From Kemlo.Rogerson <@t> waht.swest.nhs.uk Thu Nov 1 02:37:53 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu Nov 1 02:38:02 2007 Subject: [Histonet] perfusion fixation Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F099@wahtntex2.waht.swest.nhs.uk> "I don't agree. If the perfusion pressure does not match physiological pressure at least, you will not wash out all the blood. Fixative will reach some places much later than others. Red blood cells that autoflouresce and catalyze HRP reactions will remain behind. To match physiological pressure, you would need about 3 meters between the subject and the fluid reservoir. Do the math. I use double phyisological pressure, and get excellent perfusions free of red blood cells and with no damage. EM and Nissl sections look normal. Charles" Anyway, you just need a longer tube and longer arms then! The logic that you don't need a pump still stands. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From Kemlo.Rogerson <@t> waht.swest.nhs.uk Thu Nov 1 02:59:11 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu Nov 1 02:59:17 2007 Subject: [Histonet] Flammable and acid storage Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F0A1@wahtntex2.waht.swest.nhs.uk> Don't flammables and acids form esters? Knew a girl called Ester, she was pretty inflammatory. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From flip-flop <@t> ngs.ru Thu Nov 1 05:25:28 2007 From: flip-flop <@t> ngs.ru (dimas) Date: Thu Nov 1 05:25:41 2007 Subject: [Histonet] (no subject) Message-ID: please unsubscribe me From Anna.Inman <@t> stmarygj.org Thu Nov 1 10:13:50 2007 From: Anna.Inman <@t> stmarygj.org (Inman, Anna) Date: Thu Nov 1 10:14:11 2007 Subject: [Histonet] OSCAR / MNF116 ?? In-Reply-To: Message-ID: <2925AE271EAAD440AF48FCCEB8002D090542F915@smgmail01.smgj.sclhs.net> I am looking for information on two IHC antibodies. If anyone is using OSCAR or MNF116 - * Where do you purchase them from? * Do you perform it in conjunction with pankeratin? Thank you for your insight. Anna Inman SMH Pathology Anna.Inman@stmarygj.org CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From gu.lang <@t> gmx.at Thu Nov 1 11:07:00 2007 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Thu Nov 1 11:07:12 2007 Subject: [Histonet] pemphigus - ihc Message-ID: <000d01c81ca1$3d104d10$6412a8c0@dielangs.at> Hi histonetters, Has anyone experience with detecting immunoglobulins in human skin by "regular" immunohistochemistry instead of immunofluorescence? We think about a diploma thesis, that deals with the comparison of this techniques (IF on frozens and IHC on FFPE). Gudrun Lang From rgarhart <@t> system1.net Thu Nov 1 12:31:38 2007 From: rgarhart <@t> system1.net (Robert Garhart) Date: Thu Nov 1 12:31:29 2007 Subject: [Histonet] OPPORTUNITY Histology Manager Position in GA In-Reply-To: <000201c77d29$5ef145f0$800aa8c0@domain.local> References: <000201c77d29$5ef145f0$800aa8c0@domain.local> Message-ID: There is a position for a histology manager in GA. There will be two histology supervisors who will report to you. This is a great opportunity for a Histology Supervisor who would like to spend less time on the bench and more time developing technologists, working with policy & procedure and creating an efficient histology dept. Must have 5 years experience as histology manager. Please contact Robert or send resume to discuss more details. Robert Garhart Executive Recruiter System 1 Search 678-342-9029 Office rgarhart@system1.net Website: www.system1.net From jennifer.harvey <@t> Vanderbilt.Edu Thu Nov 1 13:34:56 2007 From: jennifer.harvey <@t> Vanderbilt.Edu (Harvey, Jennifer Lynn) Date: Thu Nov 1 13:35:00 2007 Subject: [Histonet] Bioloid paraffin Message-ID: Hi, Can you still buy Bioloid paraffin? It was sold by a company called Van Water and Roger Scientific, Rochester, NY. If not, is there something similar? Thanks Jennifer Harvey, HT(ASCP) QIHC Vanderbilt Vision Research Center RM 8105 MCE North Tower 1215 21st Ave. South Nashville, TN 37232-8808 Phone: 615-936-1486 From stamptrain <@t> yahoo.com Thu Nov 1 14:19:33 2007 From: stamptrain <@t> yahoo.com (Roger Moretz) Date: Thu Nov 1 14:19:37 2007 Subject: [Histonet] Staining tissue in Embed 812 resin with Hematoxylin and Eosin? In-Reply-To: <472705D9020000DB00001B4F@CE08A07M.ad.us.pri.wyeth.com> Message-ID: <113238.46931.qm@web55807.mail.re3.yahoo.com> It is not trivial, but is not impossible. A quick Google search of the terms "staining epoxy sections" turned up over 600K responses. A search of PubMed might be more to the point. The first paper I saw referenced use of H2O2 to facilitate H&E staining. Have not tried it myself. References I was more familiar with used either sodium methoxide or ethoxide to remove the plastic. Staining was pretty much straightforward after that. These techniques involve hazardous chemicals. A more useful method would be to use one of the polychromatic stains also found in the Google search I mentioned at the outset. Sorry, but I can't provide specific references--those are long gone along with the job. Roger Moretz, Ph.D. Electron Microscopist (ret.) "I'm not dead yet" Monty Python, Search for the Holy Grail. --- Lawrence Mason wrote: > I have some archived mouse tissues in Embed 812 > resin. Is it possible to stain these tissues with > hematoxylin and eosin? I was unable to find any > relevant info in the archives. > > Thanks, > Larry Mason > Wyeth Research > Cambridge, MA > _______________________________________________ > 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 afarias <@t> dmioptical.com Thu Nov 1 14:42:18 2007 From: afarias <@t> dmioptical.com (Ali Farias) Date: Thu Nov 1 14:42:39 2007 Subject: [Histonet] Leider Microscopes Message-ID: <000001c81cbf$506e3430$f14a9c90$@com> Anybody knows the leider microscope, and their website From akemiat3377 <@t> yahoo.com Thu Nov 1 14:46:30 2007 From: akemiat3377 <@t> yahoo.com (Akemi Allison-Tacha) Date: Thu Nov 1 14:46:35 2007 Subject: [Histonet] OSCAR / MNF116 ?? In-Reply-To: <2925AE271EAAD440AF48FCCEB8002D090542F915@smgmail01.smgj.sclhs.net> Message-ID: <823155.20185.qm@web31310.mail.mud.yahoo.com> You can purchase OSCAR from Cell Marque (800) 665-7284 Akemi Allison-Tacha BS, HT (ASCP) HTL Client Services Manager PhenoPath Laboratories 551 N 34th St., #100 Seattle, WA 98103 (206) 374-9000 akemi@phenopath.com http://www.phenopath.com --- "Inman, Anna" wrote: > > > I am looking for information on two IHC > antibodies. > > > > If anyone is using OSCAR or MNF116 - > > * Where do you purchase them from? > > * Do you perform it in conjunction with > pankeratin? > > > > Thank you for your insight. > > > > Anna Inman > > SMH Pathology > > Anna.Inman@stmarygj.org > > > > > > CONFIDENTIALITY NOTICE: This e-mail message, > including any attachments, is for the sole use of > the intended recipient(s) and may contain > confidential and privileged information. Any > unauthorized review, use, disclosure or distribution > is prohibited. If you are not the intended > recipient, please contact the sender by reply e-mail > and destroy all copies of the original message. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Akemi Allison-Tacha, BS, HT(ASCP)HTL President Phoenix Lab Consulting & Staffing Tele: (925)788-0900 E-Mail: akemiat3377@yahoo.com From JGREWE <@t> OhioHealth.com Thu Nov 1 15:01:25 2007 From: JGREWE <@t> OhioHealth.com (JGREWE@OhioHealth.com) Date: Thu Nov 1 15:01:36 2007 Subject: [Histonet] Jacquelyn Grewe/Staff/OhioHealth is out of the office . Message-ID: I will be out of the office starting 11/01/2007 and will not return until 11/13/2007. I will return Tuesday November 13 at 11 PM and will respond to your message when I return. Thanks, Jackie From victoria.spoon <@t> bassett.org Thu Nov 1 15:36:22 2007 From: victoria.spoon <@t> bassett.org (Spoon, Victoria) Date: Thu Nov 1 15:39:08 2007 Subject: [Histonet] Signed requisitions Message-ID: <415700FC732DE14491A3E39367834F77012456EE@ex3.bassett.org> Do labs require their pathology requisitions have the doctor's signature? I see no problem with the clinic specimens submitted with a signature. But how does this work for the OR's? The surgeons "don't even see the requisitions". Any thoughts would be appreciated. Victoria Spoon Anatomic Pathology Manager Bassett Hospital NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by New York State, and Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient or have received this communication in error please contact the sender or email.security@bassett.org and destroy all copies of the original message. Thank you. From HornHV <@t> archildrens.org Thu Nov 1 15:42:03 2007 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Thu Nov 1 15:46:10 2007 Subject: [Histonet] Signed requisitions In-Reply-To: <415700FC732DE14491A3E39367834F77012456EE@ex3.bassett.org> References: <415700FC732DE14491A3E39367834F77012456EE@ex3.bassett.org> Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6317D8298C@EMAIL.archildrens.org> Our surgeon's sign the reqs. They do this before they suit up. Our req is the official order for pathology and is filed in the chart. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 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 Spoon, Victoria Sent: Thursday, November 01, 2007 3:36 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Signed requisitions Do labs require their pathology requisitions have the doctor's signature? I see no problem with the clinic specimens submitted with a signature. But how does this work for the OR's? The surgeons "don't even see the requisitions". Any thoughts would be appreciated. Victoria Spoon Anatomic Pathology Manager Bassett Hospital NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by New York State, and Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient or have received this communication in error please contact the sender or email.security@bassett.org and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sccrshlly <@t> yahoo.com Thu Nov 1 20:02:39 2007 From: sccrshlly <@t> yahoo.com (Shelly Coker) Date: Thu Nov 1 20:02:46 2007 Subject: [Histonet] Re: paraffin and mw Message-ID: <974142.44645.qm@web90303.mail.mud.yahoo.com> Jennifer, The best option when microwave processing is keeping the paraffin in a convection oven, like one you would dry slides in. If you are reusing the paraffin, it needs to reach a temperature of more than 70 degrees celsius for the alcohol residual to evaporate off. If you are using fresh paraffin each time (very costly with high volumes and not necessary), then another option would be to use a standard paraffin pot to keep the paraffin molten. Good luck, Shelly __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From CarneyK <@t> allkids.org Fri Nov 2 07:55:07 2007 From: CarneyK <@t> allkids.org (Carney, Kathy-Jean) Date: Fri Nov 2 07:55:00 2007 Subject: [Histonet] test Message-ID: Testing service ============================================================================== CONFIDENTIALITY NOTICE: This E-mail message, including any attachments, is for the sole use of intended recipient(s)and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply E-mail and destroy all copies of the original message and its attachments without reading or saving in any manner. Thank you ============================================================================== From mlm11 <@t> cornell.edu Fri Nov 2 08:05:12 2007 From: mlm11 <@t> cornell.edu (Mary Lou Norman) Date: Fri Nov 2 08:05:16 2007 Subject: [Histonet] auto technicon locking bar Message-ID: <6.2.1.2.2.20071102085736.01eee090@postoffice9.mail.cornell.edu> Hi, To those of you who use or have used the Auto Technicon, is there a way to tighten the locking bar? Mine keeps popping open. From the February 12, 2007 Advance magazine, is anybody familiar with Julie Fuimano, an executive coach with Nurturing Your Success? Thank you, Mary Lou Norman From Kemlo.Rogerson <@t> waht.swest.nhs.uk Fri Nov 2 08:19:52 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Fri Nov 2 08:19:59 2007 Subject: [Histonet] test Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F0B6@wahtntex2.waht.swest.nhs.uk> "Testing service" Kathy-Jean You mean Weston's bus service, very testing; either none or they come all at once. How did you know? Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From Herrick.James <@t> mayo.edu Fri Nov 2 09:28:49 2007 From: Herrick.James <@t> mayo.edu (Herrick, James L.) Date: Fri Nov 2 09:28:58 2007 Subject: FW: [Histonet] Deplastification and immuno question Message-ID: <572057D3BDD52A46BD05BC6DA50686110CAEE1@MSGEBE22.mfad.mfroot.org> -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Herrick, James L. Sent: Wednesday, October 31, 2007 4:43 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Deplastification and immuno question Hi all, Has anyone had any success with attaching GMA or MMA embedded sections to glass slides, deplastifying and running immuno, without the sections loosening or falling off of the slide? If so, were you successful with antigen retrieval and antibody attachment? Thank you much for your help. Jim _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Nov 2 11:09:38 2007 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Nov 2 11:09:54 2007 Subject: [Histonet] Multiple controls Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD32048F4106@sjhaexc02.sjha.org> Will those who embed multiple type tissue controls (small pieces of several type control tissues in same block), please contact me privately? Thanks much! Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From AGrobe2555 <@t> aol.com Fri Nov 2 11:57:09 2007 From: AGrobe2555 <@t> aol.com (AGrobe2555@aol.com) Date: Fri Nov 2 11:57:18 2007 Subject: [Histonet] Re: Multiple controls Message-ID: Can you provide an e-mail address? Albert ************************************** See what's new at http://www.aol.com From portera <@t> msu.edu Fri Nov 2 12:20:55 2007 From: portera <@t> msu.edu (Amy Porter) Date: Fri Nov 2 12:23:05 2007 Subject: [Histonet] Feline retina whole mounts - tissue loss Message-ID: <002401c81d74$b9eabe00$8e7a0923@histolab> Help....I have a researcher who for years has used the same sample preparation by placing whole mount feline retinas on 1% Gelatin subbed slides and then vapor (curing) fixing with 37%-40% for 48 hours. They are suddenly having problems with tissue falling off of their slides. Does anyone out there have a suggestion or a source for some type of commercial subbing adhesive solution? Thanks in advance for the help. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 Fax: (517) 432-1368 Email: portera@msu.edu Web: www.humanpathology.msu.edu From mauger <@t> email.chop.edu Fri Nov 2 13:00:30 2007 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Fri Nov 2 13:00:59 2007 Subject: [Histonet] Xmatrix vs. Bondmax Message-ID: Hi everyone, Would anyone be willing to share their opinions on the BioGenex Xmatrix stainer? I have been demo-ing the Bondmax from Visionsbiosystems(Leica), and I am very impressed. I wonder how the xmatrix compares. It sounds much more open, and it does IF as well as IHC, FISH,ISH, and special stains!Sounds too good. Does it use disposable pipette tips? Thanks in advance. Jo Mauger From Ronald.Houston <@t> nationwidechildrens.org Fri Nov 2 13:50:33 2007 From: Ronald.Houston <@t> nationwidechildrens.org (Houston, Ronald) Date: Fri Nov 2 13:51:11 2007 Subject: [Histonet] GI dysmotility Message-ID: <979FF5962E234F45B06CF0DB7C1AABB2131B435A@chi2k3ms01.columbuschildrens.net> Is anyone aware of studies using IHC in pediatric GI dysmotility syndromes, apart form CD117 for Cajal cells? Ronnie Houston, MS, HT(ASCP)QIHC Anatomic Pathology Manager Nationwide Children's Hospital 700 Children's Drive Columbus, OH 43205 (614) 722 5465 Ronald.Houston@NationwideChildrens.org Columbus Children's Hospital is now Nationwide Children's Hospital www.NationwideChildrens.org From Ronald.Houston <@t> nationwidechildrens.org Fri Nov 2 14:23:56 2007 From: Ronald.Houston <@t> nationwidechildrens.org (Houston, Ronald) Date: Fri Nov 2 14:24:25 2007 Subject: [Histonet] GI dysmotility Message-ID: <979FF5962E234F45B06CF0DB7C1AABB2131B43FC@chi2k3ms01.columbuschildrens.net> Is anyone aware of studies using IHC in pediatric GI dysmotility syndromes, apart form CD117 for Cajal cells? Ronnie Houston, MS, HT(ASCP)QIHC Anatomic Pathology Manager Nationwide Children's Hospital 700 Children's Drive Columbus, OH 43205 (614) 722 5465 Ronald.Houston@NationwideChildrens.org Columbus Children's Hospital is now Nationwide Children's Hospital www.NationwideChildrens.org From akemiat3377 <@t> yahoo.com Fri Nov 2 14:27:23 2007 From: akemiat3377 <@t> yahoo.com (Akemi Allison-Tacha) Date: Fri Nov 2 14:27:27 2007 Subject: [Histonet] auto technicon locking bar In-Reply-To: <6.2.1.2.2.20071102085736.01eee090@postoffice9.mail.cornell.edu> Message-ID: <93100.90884.qm@web31305.mail.mud.yahoo.com> Hi, My God, you this have a autotechnicon! Well, one of the few people that can answer your question is still around! Ed Brock used to sell and demo autotechnicon's and can most likely answer your questions. He has long since retired, but he is a friend and I have his e-mail address. It is broed@aol.com I am sure he would be more than happy to help. Akemi Allison-Tacha BS, HT (ASCP) HTL Client Services Manager PhenoPath Laboratories 551 N 34th St., #100 Seattle, WA 98103 (206) 374-9000 akemi@phenopath.com http://www.phenopath.com --- Mary Lou Norman wrote: > Hi, > > To those of you who use or have used the Auto > Technicon, is there a way to > tighten the locking bar? Mine keeps popping open. > > From the February 12, 2007 Advance magazine, is > anybody familiar with > Julie Fuimano, an executive coach with Nurturing > Your Success? > > Thank you, > Mary Lou Norman > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Akemi Allison-Tacha, BS, HT(ASCP)HTL President Phoenix Lab Consulting & Staffing Tele: (925)788-0900 E-Mail: akemiat3377@yahoo.com From Maria.Mejia <@t> ucsf.edu Fri Nov 2 15:44:50 2007 From: Maria.Mejia <@t> ucsf.edu (Mejia, Maria) Date: Fri Nov 2 15:45:01 2007 Subject: [Histonet] Physitemp attachments for sliding microtome Message-ID: <6CF686BD6F24A546B85B24FE3B97864701B78FA3@EXVS06.net.ucsf.edu> Hello, If anyone out there is thinking of buying a Physitemp sliding microtome freezing stage unit measuring 3x3 inches, the pump & tank attachments for your sliding microtome - please I'd like to hear before you purchase these items. Please contact me either by email or call me at the lab. Regards Maria Bartola Mejia Department of Neurosurgery UCSF SF CA 94103 Email: maria.mejia@ucsf.edu Lab: 415-514-2954 From b-frederick <@t> northwestern.edu Fri Nov 2 18:12:41 2007 From: b-frederick <@t> northwestern.edu (b-frederick@northwestern.edu) Date: Fri Nov 2 18:12:45 2007 Subject: [Histonet] Flammable and acid storage Message-ID: <20071102231241.44F067457@merle.it.northwestern.edu> somwhere,but I can't recall. did you look at the CAP regulations -it may be there. We keep them seperate and always have. Even the acids are seperate form the bases. They can and will cross react. Rusted a few hinges in a cabinet once. Bernice ==============Original message text=============== On Wed, 31 Oct 2007 1:34:20 pm CDT "Laurie Colbert" wrote: Are there any regulations that say flammables and acids cannot be stored together? Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ===========End of original message text=========== From aakrasht <@t> yahoo.com Fri Nov 2 18:17:15 2007 From: aakrasht <@t> yahoo.com (Ali A. Krasht) Date: Fri Nov 2 18:17:20 2007 Subject: [Histonet] Price listing per type of stain Message-ID: <17056.19499.qm@web50903.mail.re2.yahoo.com> Hi There I am a histotechnologist and trying to open a general Histology laboratory and I need some one to help me on the price listing per type of stain H&E, special stains and Immunoflourence and Ill really apprecial your help on this matter and thank you. Please send me the list if you want by attachment or if you know any where I can buy or a link on the web it will be greatly appreciated and thank you again. Ali __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From mariakatleba <@t> aol.com Fri Nov 2 22:12:53 2007 From: mariakatleba <@t> aol.com (mariakatleba@aol.com) Date: Fri Nov 2 22:12:59 2007 Subject: [Histonet] Microscope donation ASAP Message-ID: <8C9EBDE8EE6F446-19C-A3E8@webmail-dd05.sysops.aol.com> Hi All, Anyone have any microscopes they would be willing to donate to a grammar school in California? I will provide the name and the contact's name (the science teacher) I am helping the school by providing my histo skills- process,embed and cut blocks on plants, tissues, etc.? I collected some (ready to be thrown out) stuff andthey appreciated it.... (old gloves, expired stains, you know!) Please reply if you can donate to this school. I am willing to pay for shipping as? the school has very very bad scopes. Thank you in advance, Maria Katleba MS HT(ASCP) Pathology Dept Manager Queen of the Valley Medical Center 1000 Trancas Napa CA 94558 ________________________________________________________________________ Email and AIM finally together. You've gotta check out free AOL Mail! - http://mail.aol.com From rjbuesa <@t> yahoo.com Sat Nov 3 07:45:55 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Nov 3 07:46:06 2007 Subject: [Histonet] Price listing per type of stain In-Reply-To: <17056.19499.qm@web50903.mail.re2.yahoo.com> Message-ID: <745533.2148.qm@web61212.mail.yahoo.com> You can try several universities web-sites because they list the price they charge for many procedures. Ren? J. "Ali A. Krasht" wrote: Hi There I am a histotechnologist and trying to open a general Histology laboratory and I need some one to help me on the price listing per type of stain H&E, special stains and Immunoflourence and Ill really apprecial your help on this matter and thank you. Please send me the list if you want by attachment or if you know any where I can buy or a link on the web it will be greatly appreciated and thank you again. Ali __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From d.non <@t> verizon.net Sat Nov 3 09:41:31 2007 From: d.non <@t> verizon.net (DNon) Date: Sat Nov 3 09:42:43 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Message-ID: <000a01c81e27$a2263570$1f0aa8c0@cyto.ocml.com> Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey From rjbuesa <@t> yahoo.com Sat Nov 3 10:59:08 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Nov 3 10:59:13 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens In-Reply-To: <000a01c81e27$a2263570$1f0aa8c0@cyto.ocml.com> Message-ID: <298860.48942.qm@web61223.mail.yahoo.com> I also had that very same problem and solved it by changing to Richard Allan's modified Harris hematoxylin. Ren? J. DNon wrote: Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ 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 d.non <@t> verizon.net Sat Nov 3 14:00:21 2007 From: d.non <@t> verizon.net (DNon) Date: Sat Nov 3 14:01:32 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens References: <000a01c81e27$a2263570$1f0aa8c0@cyto.ocml.com> <00a701c81e40$b1a797f0$1b893cd1@DJ4VDH31> Message-ID: <001e01c81e4b$caf01010$1f0aa8c0@cyto.ocml.com> Smaller specimens are wrapped in Surgipath biowraps and placed in standard cassettes. Larger specimens are sectioned and placed in standard cassettes. We've never had problems with this protocol in the 10+ years I've been working here, but we're open to any suggestions. ----- Original Message ----- From: "Markus F. Meyenhofer" To: "DNon" Sent: Saturday, November 03, 2007 1:40 PM Subject: Re: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens > In what capsules are your tissues processed?? > Regards, > Markus F. Meyenhofer > Microscopy Labs > ----- Original Message ----- > From: "DNon" > To: > Sent: Saturday, November 03, 2007 10:41 AM > Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens > > > Hello Histonet, > > Over the past several weeks our lab has had problems with hematoxylin > overstaining in the mucin of our gastric specimens. The degree of > hematoxylin overstaining is alarming in the mucin of the gastric specimens > but absent or negligible in other specimens. One pathologist reviewing > the slides indicated some of the overstained specimens appeared to be > inadequately fixed. We have addressed that issue which showed some > improvement, but the problem persists. We are making changes to increase > fixation time (although the specimens now appear adequately fixed yet > retain a large measure of the overstaining), but any further suggestions > for corrective action would be much appreciated. > > We've been hesitant to increase acid alcohol clearing time because our > other specimens are staining well with the current protocol. > > We are using the following reagents and stain times: > > - Surgipath specimen containers pre-filled with 10% neutral buffered > Formalin. > - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water > washes > - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water > wash > - Surgipath Scott's Tap Water for 1 min followed by running water wash > > Sakura automated stainer set to agitate ( up and down dipping of racks ) > once every 2 seconds, which is the fastest speed available on it. > > > Dick Non > Pathology Department > Ocean County Medical Lab > Brick, New Jersey > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From sjchtascp <@t> yahoo.com Sun Nov 4 16:00:24 2007 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Sun Nov 4 16:00:30 2007 Subject: [Histonet] Seeking an HT position Message-ID: <139801.17581.qm@web38215.mail.mud.yahoo.com> Anyone have any leads on potential HT openings in So.Wisconsin or No Illinois? Thanks, Steve __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From AnthonyH <@t> chw.edu.au Sun Nov 4 16:36:30 2007 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Sun Nov 4 16:37:45 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Message-ID: The mucin staining is usually a function of the Hematoxylin pH. Lowering the pH (usually by adding acetic acid) can significantly reduce mucin staining. 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: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Sunday, 4 November 2007 1:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ 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 note 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 JMacDonald <@t> mtsac.edu Sun Nov 4 16:54:47 2007 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Sun Nov 4 16:54:52 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens In-Reply-To: <000a01c81e27$a2263570$1f0aa8c0@cyto.ocml.com> Message-ID: Have you always used the Gill Hematoxylin? Mucin will stain with the Gill formulation. Jennifer MacDonald Director, Histotechnician Training Program Mt. San Antonio College 1100 N. Grand Ave. Walnut, CA 91789 (909) 594-5611 ext. 4884 jmacdonald@mtsac.edu "DNon" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/03/2007 07:41 AM To cc Subject [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> waht.swest.nhs.uk Mon Nov 5 02:21:24 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Nov 5 02:21:48 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F0C3@wahtntex2.waht.swest.nhs.uk> I found that when haematoxylin was 'old' I used to have that problem; I made my own 'Gill 3' variant and knew when the stain was 'going off' when mucin started to stain; the cure was a new batch of 'Gill's'. I'm assuming that the haematin oxidises to oxyhaematin and maybe this stains in a different way to its reduced brother. Your differentiation time seems rather short for Gill's 3 but even then, longer differentiation rarely eradicates the problem. The change of haematoxylin by Rene sort of suggests that it's a problem with that make/ or batch and obtaining another make or batch has solved the problem without solving the problem, if you see what I mean. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From Kemlo.Rogerson <@t> waht.swest.nhs.uk Mon Nov 5 02:27:54 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Nov 5 02:28:00 2007 Subject: [Histonet] Price listing per type of stain Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F0C4@wahtntex2.waht.swest.nhs.uk> "Hi There I am a histotechnologist and trying to open a general Histology laboratory and I need some one to help me on the price listing per type of stain H&E, special stains and Immunoflourence and Ill really apprecial your help on this matter and thank you. Please send me the list if you want by attachment or if you know any where I can buy or a link on the web it will be greatly appreciated and thank you again. Ali" Odd way round! To decide on your pricing surely you must look at your running costs and Company costs and then by looking at the work you are carrying out you figure out the price (by calculating your profit margin and putting that on top of your costs). If you use someone else's prices then how do you know if you are profitable as your costs are unlikely to be the same as theirs? If they are unprofitable then so will you be, if they are making money how will you know if you are? Surely your Business Plan will have all these calculations in them in order to get funding from Banks, or friends, or even to protect your own capital? Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From lpwenk <@t> sbcglobal.net Mon Nov 5 04:10:13 2007 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Mon Nov 5 04:10:36 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens In-Reply-To: <000a01c81e27$a2263570$1f0aa8c0@cyto.ocml.com> Message-ID: <000901c81f94$0e70e640$0202a8c0@HPPav2> Check the pH of the hematoxylin. It should be 2.2-2.9. We like to keep it at about 2.4-2.6. If the pH goes above 3.0, mucin starts to stain. Add acetic acid to the hematoxylin, and bring the pH back down. (Note: have to use a pH meter. The hematoxylin stains the pH strip blue, so they don't work when having to compare color of the strip to the color on the pH range. Personal experience with a "duh" factor at the end on my part.) Check the pH of the hematoxylin right from the bottle. If the hematoxylin pH is the problem, make certain you contact the manufacturer, tell them the lot number, and explain the problem. The manufacturers usually keep samples of each lot, and and see what the problem is (poor QC on their end, rapid decomposition, improper storage, etc.). Since the 2.2-2.9 is an acceptable range, sometimes a batch/lot will arrive that is closer to the 2.9 end. As water is carried over into the hematoxylin, the water (having a higher pH), will increase the pH of the hematoxylin, causing the hematoxylin to go over the 3.0 pH side, and thus stain the mucin. It also then starts to stain the slide, too. Increasing time in the acid wash afterwards will remove some of the mucin staining. But it's better to reduce the pH of the hematoxylin. If you are using tap water to rinse, you might also want to check the pH of the tap water. Sometimes the water companies have to add other chemicals to the water supply, to take care of problems on their side (increase bacteria, inversion of the water table levels stirring up debris, etc.). So possibly your water supply is more alkaline these past couple of weeks. Thus the carry over of water into the hematoxylin is increasing the pH of the hematoxylin a lot faster than it usually does. If this is the problem, you might need to add a step of using distilled or deionized water, after the tap water and before the hematoxylin. Let us know what the problem is. It will help other techs in your area if the problem is the water pH. It will help other techs in the country, if the problem is a lot/batch of a particular brand of hematoxylin. Good luck. Peggy A. 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 DNon Sent: Saturday, November 03, 2007 10:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Mon Nov 5 04:27:36 2007 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Mon Nov 5 04:27:57 2007 Subject: [Histonet] Flammable and acid storage In-Reply-To: <20071102231241.44F067457@merle.it.northwestern.edu> Message-ID: <000a01c81f96$7bee7640$0202a8c0@HPPav2> Try OSHA. 29 CFR 1910.106, starting in section d Lots of information about storing flammables. Boils down to - have to store flammables separate from everything else. Also the NFPA Section 30 has information. Also, check the MSDS (a requirement of OSHA). They always have information about incompatabilities. And many flammables and acids say to store them separate from each other. Peggy A. 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 b-frederick@northwestern.edu Sent: Friday, November 02, 2007 7:13 PM To: laurie.colbert@huntingtonhospital.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Flammable and acid storage somwhere,but I can't recall. did you look at the CAP regulations -it may be there. We keep them seperate and always have. Even the acids are seperate form the bases. They can and will cross react. Rusted a few hinges in a cabinet once. Bernice ==============Original message text=============== On Wed, 31 Oct 2007 1:34:20 pm CDT "Laurie Colbert" wrote: Are there any regulations that say flammables and acids cannot be stored together? Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ===========End of original message text=========== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From xqzhai <@t> uclan.ac.uk Mon Nov 5 05:25:55 2007 From: xqzhai <@t> uclan.ac.uk (Xiao Qun Zhai) Date: Mon Nov 5 05:26:47 2007 Subject: [Histonet] about Reference Manager Message-ID: <472EFDC40200009700001CD5@gwise-gw12.uclan.ac.uk> Hello everyone, This question is about Reference Manger 11. I am currently use it for my phd thesis. But I have some problems about inserting reference. Could anybody help? Is there any web forum I can discuss my questions? Thanks for help. Sunny Zhai From cmiller <@t> physlab.com Mon Nov 5 08:03:51 2007 From: cmiller <@t> physlab.com (Cheri Miller) Date: Mon Nov 5 08:03:49 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens In-Reply-To: <298860.48942.qm@web61223.mail.yahoo.com> References: <000a01c81e27$a2263570$1f0aa8c0@cyto.ocml.com> <298860.48942.qm@web61223.mail.yahoo.com> Message-ID: <013701c81fb4$b1e07230$3402a8c0@plab.local> We had the same issue and Richard Allen hematoxylin solved it as well Cheri Miller HT ASCP Histology Supervisor Physicians Laboratory Services, Inc. Omaha, NE 68117 402 738 5052 PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Saturday, November 03, 2007 10:59 AM To: DNon; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I also had that very same problem and solved it by changing to Richard Allan's modified Harris hematoxylin. Ren? J. DNon wrote: Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ 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 PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. From MSHERWOOD <@t> PARTNERS.ORG Mon Nov 5 08:08:42 2007 From: MSHERWOOD <@t> PARTNERS.ORG (Sherwood, Margaret ) Date: Mon Nov 5 08:08:47 2007 Subject: [Histonet] Flammable and acid storage In-Reply-To: <000a01c81f96$7bee7640$0202a8c0@HPPav2> Message-ID: <073AE2BEA1C2BA4A8837AB6C4B943D97B11CED@PHSXMB30.partners.org> Just another note: acids and bases need to be stored separately. We keep our acids under our hood in lab, bases in another cabinet and flammables, of course, in a yellow flammable safety cabinet. Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114-2696 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-1206 (fax) msherwood@partners.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Lee & Peggy Wenk Sent: Monday, November 05, 2007 5:28 AM To: b-frederick@northwestern.edu; laurie.colbert@huntingtonhospital.com Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Flammable and acid storage Try OSHA. 29 CFR 1910.106, starting in section d Lots of information about storing flammables. Boils down to - have to store flammables separate from everything else. Also the NFPA Section 30 has information. Also, check the MSDS (a requirement of OSHA). They always have information about incompatabilities. And many flammables and acids say to store them separate from each other. Peggy A. 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 b-frederick@northwestern.edu Sent: Friday, November 02, 2007 7:13 PM To: laurie.colbert@huntingtonhospital.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Flammable and acid storage somwhere,but I can't recall. did you look at the CAP regulations -it may be there. We keep them seperate and always have. Even the acids are seperate form the bases. They can and will cross react. Rusted a few hinges in a cabinet once. Bernice ==============Original message text=============== On Wed, 31 Oct 2007 1:34:20 pm CDT "Laurie Colbert" wrote: Are there any regulations that say flammables and acids cannot be stored together? Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ===========End of original message text=========== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted in this electronic communication is intended only for the person or entity to whom 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 information in error, please contact the Compliance HelpLine at 800-856-1983 and properly dispose of this information. From mcauliff <@t> umdnj.edu Mon Nov 5 08:21:07 2007 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Mon Nov 5 08:21:24 2007 Subject: [Histonet] about Reference Manager In-Reply-To: <472EFDC40200009700001CD5@gwise-gw12.uclan.ac.uk> References: <472EFDC40200009700001CD5@gwise-gw12.uclan.ac.uk> Message-ID: <472F26D3.5010509@umdnj.edu> Contact their tech support. Geoff Xiao Qun Zhai wrote: > Hello everyone, > > This question is about Reference Manger 11. I am currently use it for my phd thesis. But I have some problems about inserting reference. Could anybody help? Is there any web forum I can discuss my questions? > > Thanks for help. > > Sunny Zhai > > _______________________________________________ > 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 mcauliff@umdnj.edu ********************************************** From pierre.chaumat <@t> alphelys.com Mon Nov 5 08:29:11 2007 From: pierre.chaumat <@t> alphelys.com (Pierre CHAUMAT) Date: Mon Nov 5 08:29:20 2007 Subject: [Histonet] Tissue fixative FIXALL Message-ID: <0MKwh2-1Ip2ww1uLo-0004zH@mrelayeu.kundenserver.de> Dear members, Any one has heard or even use FIXALL tissue fixative from Carlo Erba ? Best regards Pierre Pierre CHAUMAT President & CEO ALPHELYS SAS Impasse Paul Langevin Ferme des Ebisoires F-78370 PLAISIR FRANCE Tel +33 1 30 07 52 95 Fax +33 1 30 07 51 56 Cell +33 6 03 47 75 92 http://www.alphelys.com ********************************************************************** This e-mail and any files transmitted with it are confidential and intended solely for the use of the individual to whom it is addressed. If you have received this email in error please send it back to the person that sent it to you. Any views or opinions presented are solely those of its author and do not necessarily represent those of ALPHELYS SAS or any of its subsidiary companies. Unauthorized publication, use, dissemination, forwarding, printing or copying of this email and its associated attachments is strictly prohibited. ********************************************************************** From Traczyk7 <@t> aol.com Mon Nov 5 10:47:16 2007 From: Traczyk7 <@t> aol.com (Traczyk7@aol.com) Date: Mon Nov 5 10:47:26 2007 Subject: [Histonet] Histologist of the Year Message-ID: Peggy: Thank you for taking the time to answer questions posted on HistoNet. The information you share, and the obvious thought behind it, is very much appreciated. Your recent award from NSH for Histologist of the Year was well deserved. Congratulations. Dorothy Dorothy Murphy Traczyk Hacker Instruments & Industries Inc. ************************************** See what's new at http://www.aol.com From oshel1pe <@t> cmich.edu Mon Nov 5 07:23:35 2007 From: oshel1pe <@t> cmich.edu (Philip Oshel) Date: Mon Nov 5 10:52:54 2007 Subject: [Histonet] Flammable and acid storage In-Reply-To: <20071102231241.44F067457@merle.it.northwestern.edu> References: <20071102231241.44F067457@merle.it.northwestern.edu> Message-ID: I don't know if CAP covers chemical storage, but in a sense it doesn't matter. In the US, the federal OSHA does, and in this case OSHA is who matters. By OSHA regs, flammables must be stored in a properly built and labeled flammable-chemical locker, and acids and bases stored separately from flammables and from each other, and are supposed to be in specially made and labeled storage lockers, although my experience with acids and bases is that special lockers are absolute requirements, if the amounts aren't large and the chemicals are stored properly and separately. Further, these storage cabinets are supposed to be individually vented to the outside of the building -- at least, the flammables locker is. Again, this regulation doesn't seem to be an absolute. There are more requirements I don't recall off hand, but your institution's safety people should have those regs. OSHA can, will, and has shut down and fined facilities that violate chemical storage and handling regulations (we just had a visit from the state version, no problems that I heard of, but ...). By "facilities", I don't mean industrial or commercial, but colleges, hospitals, and the like. Phil >somwhere,but I can't recall. did you look at the CAP regulations -it >may be there. We keep them >seperate and always have. Even the acids are seperate form the >bases. They can and will cross >react. Rusted a few hinges in a cabinet once. >Bernice > >==============Original message text=============== >On Wed, 31 Oct 2007 1:34:20 pm CDT "Laurie Colbert" wrote: > >Are there any regulations that say flammables and acids cannot be stored >together? > > > >Laurie Colbert > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >===========End of original message text=========== -- Philip Oshel Microscopy Facility Supervisor Biology Department 024C Brooks Hall Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 From asmith <@t> mail.barry.edu Sun Nov 4 11:31:04 2007 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Mon Nov 5 10:54:02 2007 Subject: [Histonet] Flammable and acid storage In-Reply-To: <20071102231241.44F067457@merle.it.northwestern.edu> Message-ID: <7DBCCC1FBC77C94F99F920D0CA6400B61E4139@exchsrv01.barrynet.barry.edu> Regardless of regulations, nitric, sulfuric, and perchloric acids should be kept separate from organic compounds: they can form very dangerous reaction products. Hydrochloric acid is best kept with the other mineral acids; although it can react nastily with nitric acid, it also reacts nastily with formaldehyde. For storage, formic and acetic acids should be treated as organic compounds rather than acids: they can react violently with oxidizing mineral acids; their organic reactions are gentler. I like to keep my acids in high-density polyethylene cabinets; hydrochloric and nitric acids tend to rust out the metal ones, and a drop of sulfuric acid running down the side of the bottle can make an unholy mess of a wooden cabinet. 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 b-frederick@northwestern.edu Sent: Friday, November 02, 2007 6:13 PM To: laurie.colbert@huntingtonhospital.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Flammable and acid storage somwhere,but I can't recall. did you look at the CAP regulations -it may be there. We keep them seperate and always have. Even the acids are seperate form the bases. They can and will cross react. Rusted a few hinges in a cabinet once. Bernice ==============Original message text=============== On Wed, 31 Oct 2007 1:34:20 pm CDT "Laurie Colbert" wrote: Are there any regulations that say flammables and acids cannot be stored together? Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ===========End of original message text=========== _______________________________________________ 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 bakerj <@t> umich.edu Mon Nov 5 11:07:07 2007 From: bakerj <@t> umich.edu (John Baker) Date: Mon Nov 5 11:09:14 2007 Subject: [Histonet] deplasticizing PMMA sections for staining Message-ID: <26b16e62cc93c54c84205288e3868fe5@umich.edu> > > > Hello All, We are sectioning PMMA embedded rat femur at 5 microns on > the Polycut. Then we are deplasticizing for H & E and Trichrome > stains. It has been taking over 2 weeks with several changes of xylene > to get them done and still some remnants of plastic there. I see in > Sheehan's Practice and Theory of Histotechnology that in several > protocols they deplasticize in xylene for 4 hours at 55 degrees C. > The flash point of xylene is 25 degree C. Is it safe in covered > staining dishes to actually do this? Also, we use Mayer's Hematoxylin > and 2% Eosin to stain. What H & E protocol do you use for bone? Any > suggestions on how to speed up the deplasticizing process welcome. > Does anyone have Diane Sterchi's and Gayle Callis's email addresses, > lost them? > Thanks, John > > > > > John A. Baker The University of Michigan Orthopaedic Research Laboratories Histology Unit 109 Zina Pitcher Place, 2218 BSRB Ann Arbor, MI 48109-2200 734-936-1635 From hlukey <@t> msn.com Mon Nov 5 12:22:56 2007 From: hlukey <@t> msn.com (Hugh Luk) Date: Mon Nov 5 12:23:04 2007 Subject: [Histonet] TMA arrayers. Does anyone have a favorite model? Message-ID: I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts!? Play Star Shuffle:? the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct From nancy.troiano <@t> yale.edu Mon Nov 5 12:25:52 2007 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Mon Nov 5 12:25:58 2007 Subject: [Histonet] Deplasticizing MMA sections Message-ID: <5.2.1.1.2.20071105132352.00c3f668@email.med.yale.edu> Try soaking the slides in two 5 minute changes of acetone (100%) to deplastify or you can deplastify in 2 changes of Cellosolve (Fisher, cat. #E181-4) 25 minutes each, followed by 5 minutes in 70% ethanol, 40% ethanol, then distilled water for five minutes prior to doing your staining. From Xorren <@t> aol.com Mon Nov 5 12:44:13 2007 From: Xorren <@t> aol.com (Xorren@aol.com) Date: Mon Nov 5 12:44:31 2007 Subject: [Histonet] Histo, cyto job in NY Message-ID: Hello all ! Does anyone know of any histology or cytology prep job in the NY area. I have have 15 years in the field and hope someone has some info. you can respond @ xorren@aol.com or call (516) 671-0907 have a great day !!!!! ************************************** See what's new at http://www.aol.com From gvdobbin <@t> ihis.org Mon Nov 5 12:44:33 2007 From: gvdobbin <@t> ihis.org (Greg Dobbin) Date: Mon Nov 5 12:44:50 2007 Subject: [Histonet] TMA arrayers. Does anyone have a favorite model? Message-ID: I am investigating an instrument that I saw at NSH by RMC Products. Model no. is KIN-1. The picture on the web site does not match exactly the unit they had at NSH but they could send you the same broshure they were passing out at their booth (it must be available as a pdf). Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> hlukey@msn.com 11/5/2007 2:22 PM >>> I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. From sbreeden <@t> nmda.nmsu.edu Mon Nov 5 12:50:55 2007 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Mon Nov 5 12:51:06 2007 Subject: [Histonet] NSH Report Message-ID: <4D14F0FC9316DD41972D5F03C070908B8F4725@nmdamailsvr.nmda.ad.nmsu.edu> Okay, I'm back from my week at NSH and this is my report. 1. The NSH staff is awesome. Aubrey, Carrie and the entire NSH staff pulled off yet another flawless meeting. We can't thank NSH enough for all they do! 2. Joe Nocito is alive and well. I met him. He lives and he wasn't wearing a flak jacket. 3. Our vendors are magnificent - supportive, creative, helpful and approachable. Kudos!! 4. Each one of us needs to recruit NSH members - there is strength in numbers. NSH is our connection in SO many ways! 5. Ada Feldman makes a terrific clown! 6. Did you know that histologists are the ONLY technicians not recognized by the US Government as healthcare professionals? 7. After 3 days of walking the 6 blocks from hotel to convention center, I discovered the free bus. 8. There must be a "message board" at next year's meeting so we can find each other if we need to. 9. I want to know what vitamins Peggy Wenk takes. Seriously. Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 From tp2 <@t> medicine.wisc.edu Mon Nov 5 12:55:36 2007 From: tp2 <@t> medicine.wisc.edu (Thomas Pier) Date: Mon Nov 5 12:56:01 2007 Subject: [Histonet] TMA arrayers. Does anyone have a favorite model? Message-ID: <472F12C8020000DF00002B3D@gwmail.medicine.wisc.edu> I haven't used any other instruments, but I've been happy with the Beecher MTA-1. Tom Pier >>> Hugh Luk 11/05/07 12:22 PM >>> I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Boneslides <@t> aol.com Mon Nov 5 13:10:40 2007 From: Boneslides <@t> aol.com (Boneslides@aol.com) Date: Mon Nov 5 13:11:23 2007 Subject: [Histonet] Re: Histonet Digest, Vol 48, Issue 5 Message-ID: I de-plasticize my PMMA sections in methyl acetate...takes about 30 minutes, then rehydrate thru several changes of absolute, 95% and 70% alcohol, rinse in water, stain as desired. Diane M. Mahovlic, HT, MLT(ASCP) Orthopedic Pathology & Biomaterials Laboratory Department of Anatomic Pathology The Cleveland Clinic Foundation 9500 Euclid Avenue- L30 Cleveland, Ohio 44195 216-444-0166 ************************************** See what's new at http://www.aol.com From PMonfils <@t> Lifespan.org Mon Nov 5 13:22:08 2007 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Nov 5 13:22:15 2007 Subject: [Histonet] deplasticizing PMMA sections for staining In-Reply-To: <26b16e62cc93c54c84205288e3868fe5@umich.edu> Message-ID: <4EBFF65383B74D49995298C4976D1D5E273CF6@LSRIEXCH1.lsmaster.lifespan.org> In my experience, xylene is a rather poor solvent for PMMA. It works much faster at 60 degrees, but don't try heating flammable solvents in your oven unless you are certain it is certified explosion-proof (most paraffin ovens are not). Even then, unless you have the oven in or at least next to a fume hood, you'll get a lot of xylene vapors in the air as a result of heating it. I deplasticise PMMA with 2-methoxyethyl acetate, which only takes a couple of hours at room temperature. This has a rather strong though not unpleasant odor, and should be used in the hood. From gvdobbin <@t> ihis.org Mon Nov 5 14:12:32 2007 From: gvdobbin <@t> ihis.org (Greg Dobbin) Date: Mon Nov 5 14:12:54 2007 Subject: [Histonet] TMA arrayers. Web link Message-ID: Sorry. I see the company name is actually Boeckeler and the website is www.boeckeler.com Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> "Greg Dobbin" 11/5/2007 2:44 PM >>> I am investigating an instrument that I saw at NSH by RMC Products. Model no. is KIN-1. The picture on the web site does not match exactly the unit they had at NSH but they could send you the same broshure they were passing out at their booth (it must be available as a pdf). Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> hlukey@msn.com 11/5/2007 2:22 PM >>> I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. From trathborne <@t> somerset-healthcare.com Mon Nov 5 14:30:49 2007 From: trathborne <@t> somerset-healthcare.com (Rathborne, Toni) Date: Mon Nov 5 14:31:13 2007 Subject: [Histonet] NSH show Message-ID: How was the show? I heard there were a lot of vendors there. What about the lectures/workshops? I was hoping to go, so many relevant topics offered. A friend of mine was there and mentioned that Ventana had some sort of secretive booth. What was that all about? Is it a new IHC stainer? Or part of their whole "lab platform"? Toni CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. Somerset Medical Center is proud to receive the Somerset County Business Partnership's 2006 Quality of Life Award. From Charles.Embrey <@t> carle.com Mon Nov 5 15:08:54 2007 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Mon Nov 5 15:10:11 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens In-Reply-To: <000a01c81e27$a2263570$1f0aa8c0@cyto.ocml.com> Message-ID: <44780C571F28624DBB446DE55C4D733A1FE565@EXCHANGEBE1.carle.com> I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bwhitaker <@t> brownpathology.com Mon Nov 5 15:22:45 2007 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Mon Nov 5 15:19:12 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens In-Reply-To: <44780C571F28624DBB446DE55C4D733A1FE565@EXCHANGEBE1.carle.com> Message-ID: <003501c81ff2$02602060$3601a8c0@brownpathology.net> Hi All, I had the same type incident (with the increase in mucin staining with the Surgipath hematoxylin), but it was a one time thing that we attributed to a larger than normal volume of H&E's in that particular time period. We increased the frequency that solutions are changed slightly, and haven't had another problem. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, November 05, 2007 3:09 PM To: DNon; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@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 doug <@t> ppspath.com Mon Nov 5 15:26:45 2007 From: doug <@t> ppspath.com (Douglas D Deltour) Date: Mon Nov 5 15:27:21 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens In-Reply-To: <44780C571F28624DBB446DE55C4D733A1FE565@EXCHANGEBE1.carle.com> Message-ID: Charles, We also are having the same issue with the Surgipath Gills III. This issue also started last month. Very interesting. Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, November 05, 2007 4:09 PM To: DNon; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@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 ryaskovich <@t> dir.nidcr.nih.gov Mon Nov 5 15:43:34 2007 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR) [E]) Date: Mon Nov 5 15:43:39 2007 Subject: [Histonet] deplasticizing PMMA sections for staining In-Reply-To: <26b16e62cc93c54c84205288e3868fe5@umich.edu> References: <26b16e62cc93c54c84205288e3868fe5@umich.edu> Message-ID: John, You can soak them in 3 changes of acetone for 15 minutes each. Put them in the first acetone right out of the oven. Ruth Yaskovich National Institutes of Health National Institute of Dental and Craniofacial Research -----Original Message----- From: John Baker [mailto:bakerj@umich.edu] Sent: Monday, November 05, 2007 12:07 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] deplasticizing PMMA sections for staining > > > Hello All, We are sectioning PMMA embedded rat femur at 5 microns on > the Polycut. Then we are deplasticizing for H & E and Trichrome > stains. It has been taking over 2 weeks with several changes of xylene > to get them done and still some remnants of plastic there. I see in > Sheehan's Practice and Theory of Histotechnology that in several > protocols they deplasticize in xylene for 4 hours at 55 degrees C. > The flash point of xylene is 25 degree C. Is it safe in covered > staining dishes to actually do this? Also, we use Mayer's Hematoxylin > and 2% Eosin to stain. What H & E protocol do you use for bone? Any > suggestions on how to speed up the deplasticizing process welcome. > Does anyone have Diane Sterchi's and Gayle Callis's email addresses, > lost them? > Thanks, John > > > > > John A. Baker The University of Michigan Orthopaedic Research Laboratories Histology Unit 109 Zina Pitcher Place, 2218 BSRB Ann Arbor, MI 48109-2200 734-936-1635 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From b-frederick <@t> northwestern.edu Mon Nov 5 15:45:56 2007 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Mon Nov 5 15:46:16 2007 Subject: [Histonet] NSH Message-ID: <000001c81ff5$42a1aec0$d00f7ca5@lurie.northwestern.edu> This was my first meeting and whereas I enjoyed myself tremendously I only had 2 gripes 1. There should be a block of time set aside to see the vendors. Most of us were in workshops for a lot of the time and had to do quick run up to the vendors. 2. I was surprised water was only provided for the speakers. It was so dry I was going through 36-48 oz of water every day and at 2$ a pop... If I'd known, I'd have run off to Walgreens or somewhere. I do plan to be back and really enjoyed everybody and everything. Missed meeting Joe though I thought we had arranged for that (tee hee). Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 From ander093 <@t> tc.umn.edu Mon Nov 5 15:51:47 2007 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Mon Nov 5 15:52:06 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens In-Reply-To: <44780C571F28624DBB446DE55C4D733A1FE565@EXCHANGEBE1.carle.c om> References: <000a01c81e27$a2263570$1f0aa8c0@cyto.ocml.com> <44780C571F28624DBB446DE55C4D733A1FE565@EXCHANGEBE1.carle.com> Message-ID: I too have had problems recently with Surgipath hematoxylin in brain tissue (not mucin). They replaced what I had with a new bottle, but I am seeing the same problems. I believe the formula has been changed to eliminate mercury--don't know if this is the basis of the problems or not--but I believe that is when I started having problems. They have a "new" hematoxylin (560) and eosin (515), but our pathologists did not care for those either. I have been using Surgipath for 14 years and just started having staining problems with it. LuAnn Anderson Neuropthology Lab University of Minnesota At 03:08 PM 11/5/2007, Charles.Embrey wrote: >I find this post most interesting. After using Surgipath Gills III for >the past 7 years we started having a problem with blue mucin just last >month. I corrected the problem by adding GAA but I now have to monitor >the pH on a daily basis. It is strange that after all these years I >would suddenly have this problem and then read about another lab having >the same difficulty with Surgipath Hematoxylin starting about the same >timeframe. Has Surgipath changed something? > >Charles Embrey Jr., PA(ASCP) >Histology Manager, Carle Clinic > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon >Sent: Saturday, November 03, 2007 9:42 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens > >Hello Histonet, > > Over the past several weeks our lab has had problems with >hematoxylin overstaining in the mucin of our gastric specimens. The >degree of hematoxylin overstaining is alarming in the mucin of the >gastric specimens but absent or negligible in other specimens. One >pathologist reviewing the slides indicated some of the overstained >specimens appeared to be inadequately fixed. We have addressed that >issue which showed some improvement, but the problem persists. We are >making changes to increase fixation time (although the specimens now >appear adequately fixed yet retain a large measure of the overstaining), >but any further suggestions for corrective action would be much >appreciated. > > We've been hesitant to increase acid alcohol clearing time because >our other specimens are staining well with the current protocol. > >We are using the following reagents and stain times: > >- Surgipath specimen containers pre-filled with 10% neutral buffered >Formalin. >- Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running >water washes >- .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water >wash >- Surgipath Scott's Tap Water for 1 min followed by running water wash > >Sakura automated stainer set to agitate ( up and down dipping of racks ) >once every 2 seconds, which is the fastest speed available on it. > > >Dick Non >Pathology Department >Ocean County Medical Lab >Brick, New Jersey >_______________________________________________ >Histonet mailing list >Histonet@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 fudo <@t> pathology.ufl.edu Mon Nov 5 15:52:46 2007 From: fudo <@t> pathology.ufl.edu (FU,DONTAO) Date: Mon Nov 5 15:52:53 2007 Subject: [Histonet] how to keep thick sections on the slides after deparaffin Message-ID: <81FEA2B93DC53C4B8CF3667707CA592A0DCE8C@path-exchange.ad.ufl.edu> Hi, I met a big problem in keeping thick sections on the slides after deparaffin recently. I cut a 70um-thick mouse retina section on superfrost plus Gold slides, leaving it dry for 2days. Before deparaffin, I put the slide in the 65C oven for 30 min to let it melt, then I did the general deparaffin procedure. Unfortunately, the section fell off from the slide. Does anyone have any experience on deparaffin thick sections? Any suggestions? Thank you, Ann Dongtao Fu MD Ph.D Lab Manager Molecular Pathology core Dept. of Pathology University of Florida Lab Phone: 352-273-7752 FAX: 352-273-7753 Rm: D11-50 From d.non <@t> verizon.net Mon Nov 5 17:51:18 2007 From: d.non <@t> verizon.net (DNon) Date: Mon Nov 5 17:52:32 2007 Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens References: <44780C571F28624DBB446DE55C4D733A1FE565@EXCHANGEBE1.carle.com> Message-ID: <000601c82006$c3081ac0$1f0aa8c0@cyto.ocml.com> I spoke with Surgipath's QC department today and asked if there was a change in their Gill's III formulation. According to Mike Urban at Surgipath, nothing has changed. Nonetheless, he is going to run additional tests on the lot number in question and will get back to me. I find your post very interesting indeed as our lab has used Surgipath Gill's III for the last 10 years. They ( Surgipath) will also be sending me a gallon of Gill's III from a different lot. If it doesn't show improvement, we'll either lower the ph similar to what you did, or switch to Harris hematoxylin. ----- Original Message ----- From: "Charles.Embrey" To: "DNon" ; Sent: Monday, November 05, 2007 4:08 PM Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> waht.swest.nhs.uk Tue Nov 6 02:24:24 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Tue Nov 6 02:24:35 2007 Subject: [Histonet] how to keep thick sections on the slides after deparaffin Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F0E6@wahtntex2.waht.swest.nhs.uk> Hi, I met a big problem in keeping thick sections on the slides after deparaffin recently. I cut a 70um-thick mouse retina section on superfrost plus Gold slides, leaving it dry for 2days. Before deparaffin, I put the slide in the 65C oven for 30 min to let it melt, then I did the general deparaffin procedure. Unfortunately, the section fell off from the slide. Does anyone have any experience on deparaffin thick sections? Any suggestions? Thank you, You could use bovine albumin to coat the slides (I think 1% but I'm not sure) then dry at 37 degrees for those two days then into the oven at 60 degrees (ish). Think that sort of sets the section onto the slide but you need to get the concentration correct as the protein could stain with some techniques if its too concentrated. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From barbwebb <@t> webtv.net Tue Nov 6 04:07:04 2007 From: barbwebb <@t> webtv.net (Barbara Webb) Date: Tue Nov 6 04:07:15 2007 Subject: [Histonet] FYI - Brainbow Message-ID: Looking at the fuure? http://www.nytimes.com/2007/11/06/health/research/06brai.html?_r=1&ref=science&oref=slogin From lpwenk <@t> sbcglobal.net Tue Nov 6 04:20:07 2007 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Tue Nov 6 04:20:32 2007 Subject: [Histonet] Histo, cyto job in NY In-Reply-To: Message-ID: <000801c8205e$9b559130$0202a8c0@HPPav2> If you are looking for a job as a histotech in New York, it's not going to happen. The NY legislators have made a licensure law that only certified med techs can be histotechs. Those already working as histotechs in NY can continue to work in NY. However, any certified histotech moving into NY cannot work as a histotech until they take all the med tech courses and pass the med tech certification exam. NY will not recognize the NAACLS histotech program graduates from SUNY, the only accredited HT program in NY. (SUNY will be closing, as it's impossible to offer all med tech courses and all histotech courses, all in a 2 year program.) NY will not recognize those earning their ASCP HT or HTL certification through the associate degree on-the-job training route. ASCP, NY histotech society, and the NY pathologist groups are lobbying for changes, but the legislators are not listening. NSH has weighed in with ASCP, and the NY pathologists are coming to NSH to see what NSH can do, too. (That's where Vinnie DelaSperanza's (NSH presidents) comments on needing more members come into play. If ASCP and pathologist groups aren't swaying the legislators, then NSH with our fewer numbers are not going to help. Histotechs need to join NSH and ASCP, to histotechs have more numbers and will be listened to better.) The original draft of the bill, according to Vinnie, included histotechs with the appropriate information on training and certification and job responsibilities. The bill got changed over the years, and went unnoticed by NY histotechs. The bill took years (decade+) to pass, so the legislators don't want to change anything on the law at this stage. It may take years to correct. At this time, the med tech programs do not have classes in the theory or practical aspects of histotechnology. I don't know if they are planning on adding them to the med tech curriculum. In the mean time, NY histotechs will be retiring and leaving the field. Workloads will increase as the patient populations ages and technology requires more tissue tests. And there will be fewer and fewer NY histotechs to do these jobs. It's a histology crisis in the making. Peggy A. 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 Xorren@aol.com Sent: Monday, November 05, 2007 1:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histo, cyto job in NY Hello all ! Does anyone know of any histology or cytology prep job in the NY area. I have have 15 years in the field and hope someone has some info. you can respond @ xorren@aol.com or call (516) 671-0907 have a great day !!!!! ************************************** See what's new at http://www.aol.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From relia1 <@t> earthlink.net Tue Nov 6 07:21:20 2007 From: relia1 <@t> earthlink.net (Pam Barker) Date: Tue Nov 6 07:21:26 2007 Subject: [Histonet] RELIA's Histology Opportunities Update 11-06-07 Message-ID: Hi Histonetters! I have been reading some of the posts on the NSH in Denver. What about everybody else, did you make it to the NSH in Denver? If so how was it? What was your favorite thing about it? I haven?t been since the convention in 2005 in Ft. Lauderdale. Did you have a get together for the histonet? We did at the Ft. Lauderdale convention it was a small turnout but we had fun. I really enjoyed meeting people, learning new things and the party at the Hard Rock Casino. I am planning on attending the 2008 meeting in Pittsburgh. Will I see you there? Here is a list of my current openings all of the positions are full time permanent positions. They are M-F dayshift opportunities and my clients offer excellent compensation, benefits and relocation assistance. And they are ready to interview and hire right away. ****RELIA Spotlight Opportunities**** I am working with a client in the Philadelphia, PA area that has 2 unique and exciting opportunities. These are nonclinical positions. BioSciences Product Manager Histologist Technical and Field Support ********************************* Histology Managers/Supervisors needed in: Dallas, TX Austin, TX Philadelphia, PA Valencia, CA Cape Cod, MA Histotechnicians/Histotechnologists needed in: Phoenix, AZ Atlanta, GA San Francisco, CA Los Angeles, CA Valencia, CA Orlando, FL St. Petersburg, FL Fredericksburg, VA Dallas, TX Waco, TX Austin, TX Philadelphia, PA Pittsburgh, PA If you know of anyone else who might be interested in any of these positions or might like to subscribe to RELIA?s Histology Careers Bulletin please feel free to pass this e-mail along to them. Remember it never hurts to look!!! Thanks, Pam 877-60 RELIA (877-607-3542) Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net www.myspace.com/pamatrelia From jwatson <@t> gnf.org Tue Nov 6 08:41:31 2007 From: jwatson <@t> gnf.org (James Watson) Date: Tue Nov 6 08:43:12 2007 Subject: [Histonet] Histo, cyto job in NY References: <000801c8205e$9b559130$0202a8c0@HPPav2> Message-ID: At NSH we were told by someone that was in contact with the legistators that wrote the bill that this did not pass and that they were rewriting the bill. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Lee & Peggy Wenk Sent: Tue 11/6/2007 2:20 AM To: Xorren@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histo, cyto job in NY If you are looking for a job as a histotech in New York, it's not going to happen. The NY legislators have made a licensure law that only certified med techs can be histotechs. Those already working as histotechs in NY can continue to work in NY. However, any certified histotech moving into NY cannot work as a histotech until they take all the med tech courses and pass the med tech certification exam. NY will not recognize the NAACLS histotech program graduates from SUNY, the only accredited HT program in NY. (SUNY will be closing, as it's impossible to offer all med tech courses and all histotech courses, all in a 2 year program.) NY will not recognize those earning their ASCP HT or HTL certification through the associate degree on-the-job training route. ASCP, NY histotech society, and the NY pathologist groups are lobbying for changes, but the legislators are not listening. NSH has weighed in with ASCP, and the NY pathologists are coming to NSH to see what NSH can do, too. (That's where Vinnie DelaSperanza's (NSH presidents) comments on needing more members come into play. If ASCP and pathologist groups aren't swaying the legislators, then NSH with our fewer numbers are not going to help. Histotechs need to join NSH and ASCP, to histotechs have more numbers and will be listened to better.) The original draft of the bill, according to Vinnie, included histotechs with the appropriate information on training and certification and job responsibilities. The bill got changed over the years, and went unnoticed by NY histotechs. The bill took years (decade+) to pass, so the legislators don't want to change anything on the law at this stage. It may take years to correct. At this time, the med tech programs do not have classes in the theory or practical aspects of histotechnology. I don't know if they are planning on adding them to the med tech curriculum. In the mean time, NY histotechs will be retiring and leaving the field. Workloads will increase as the patient populations ages and technology requires more tissue tests. And there will be fewer and fewer NY histotechs to do these jobs. It's a histology crisis in the making. Peggy A. 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 Xorren@aol.com Sent: Monday, November 05, 2007 1:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histo, cyto job in NY Hello all ! Does anyone know of any histology or cytology prep job in the NY area. I have have 15 years in the field and hope someone has some info. you can respond @ xorren@aol.com or call (516) 671-0907 have a great day !!!!! ************************************** See what's new at http://www.aol.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 tissuearray <@t> hotmail.com Tue Nov 6 08:52:39 2007 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Tue Nov 6 08:52:53 2007 Subject: [Histonet] TMA arrayers. Web link In-Reply-To: References: Message-ID: Check out the "Arraymold" Go to: http:/www.arraymold.com > Date: Mon, 5 Nov 2007 16:12:32 -0400> From: gvdobbin@ihis.org> To: Histonet@lists.utsouthwestern.edu> Subject: Re: [Histonet] TMA arrayers. Web link> CC: > > Sorry. I see the company name is actually Boeckeler and the website is> www.boeckeler.com> > Greg Dobbin, R.T.> Chief Technologist, Histology Lab> Dept. of Laboratory Medicine,> Queen Elizabeth Hospital,> P.O. Box 6600> Charlottetown, PE C1A 8T5> Phone: (902) 894-2337> Fax: (902) 894-2385> > There is some merit in doing the right thing rather badly,> but absolutely none in doing the wrong thing excellently!> > >>> "Greg Dobbin" 11/5/2007 2:44 PM >>>> I am investigating an instrument that I saw at NSH by RMC Products.> Model no. is KIN-1. The picture on the web site does not match exactly> the unit they had at NSH but they could send you the same broshure> they> were passing out at their booth (it must be available as a pdf).> Cheers!> Greg> > Greg Dobbin, R.T.> Chief Technologist, Histology Lab> Dept. of Laboratory Medicine,> Queen Elizabeth Hospital,> P.O. Box 6600> Charlottetown, PE C1A 8T5> Phone: (902) 894-2337> Fax: (902) 894-2385> > There is some merit in doing the right thing rather badly,> but absolutely none in doing the wrong thing excellently!> > >>> hlukey@msn.com 11/5/2007 2:22 PM >>>> > I need advice. Is there a favorite Tissue microarray unit? I> currently use the Beecher TMA #1, but due to the humidity in Hawaii,> the> electronics have failed. I guess I shouldn't have taken it swimming> with me in the ocean! I attending NSH, but did not see too many TMA> vendors. Beecher did throw a very nice break-time pretzel party, but I> missed meeting any of their people. I would appreciate any advice...> Thanks,> Hugh Luk, HTL (ASCP)> hlukey@msn.com > or hluk@crch.hawaii.edu > Pathology Shared Resources Lab > Cancer Research Center of Hawaii > Honolulu, Hawaii> _________________________________________________________________> Climb to the top of the charts! Play Star Shuffle: the word scramble> challenge with star power.> http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct_______________________________________________> > > Histonet mailing list> Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Statement of Confidentiality> This message (including attachments) may contain confidential or> privileged information intended for a specific individual or> organization. If you have received this communication in error, please> notify the sender immediately. If you are not the intended recipient,> you are not authorized to use, disclose, distribute, copy, print or rely> on this email, and should promptly delete this email from your entire> computer system. > > D?claration de confidentialit?> Le pr?sent message (y compris les annexes) peut contenir des> renseignements confidentiels ayant pour objet une personne ou un> organisme particulier. Si vous avez re?u la pr?sente communication par> erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes> pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser,> divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous> en servir, et vous devriez l'effacer compl?tement de votre syst?me> informatique.> > _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet> > Statement of Confidentiality> This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. > > D?claration de confidentialit?> Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique.> > _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Help yourself to FREE treats served up daily at the Messenger Caf?. Stop by today. http://www.cafemessenger.com/info/info_sweetstuff2.html?ocid=TXT_TAGLM_OctWLtagline From michelle-griffin-reyes <@t> uiowa.edu Tue Nov 6 08:57:25 2007 From: michelle-griffin-reyes <@t> uiowa.edu (Griffin-Reyes, Michelle A) Date: Tue Nov 6 08:57:27 2007 Subject: [Histonet] eosinophil peroxidase (EPO) Message-ID: Does anyone have any experience using eosinophil peroxidase (EPO) to stain FFPE tissues (I am doing other staining methods as well)? I have searched and can find EPO usage for western blot and flow and one publication using frozen tissues but nothing for FFPE tissues. Any suggestions would be greatly appreciated. Thanks in advance!! Michelle A. Griffin-Reyes Comparative Pathology Laboratory University of Iowa Hospitals and Clinics 319-384-4620 From shive003 <@t> umn.edu Tue Nov 6 09:47:28 2007 From: shive003 <@t> umn.edu (Jan Shivers) Date: Tue Nov 6 09:47:34 2007 Subject: [Histonet] Coxiella burnetii control tissue Message-ID: <00a301c8208c$55e04310$a1065486@auxs.umn.edu> I'm looking for FFPE positive control tissue in order to work up an IHC test for Coxiella burnetii. Can anyone point me in the right direction of a source? Thanks in advance, Jan Shivers IHC/Histo/EM Veterinary Diagnostic Lab UMN College of Veterinary Medicine St. Paul, MN, USA shive003@umn.edu From tkngflght <@t> yahoo.com Tue Nov 6 10:38:58 2007 From: tkngflght <@t> yahoo.com (Cheryl R. Kerry) Date: Tue Nov 6 10:39:23 2007 Subject: [Histonet] NY License rules--link to the official site In-Reply-To: <000601c82006$c3081ac0$1f0aa8c0@cyto.ocml.com> Message-ID: <000001c82093$8842f530$6801a8c0@CHERYLSLAPTOP> http://www.op.nysed.gov/clp.htm http://www.op.nysed.gov/clp.htm Follow the above link and look at the clinical tech rules. I've assisted close to a dozen histotechs get licensed in the last year. If you aren't sure what the guidelines say, call them--they're pretty easy to get on the phone. This is a work in progress. If you have trouble, voice your concern to the legislature and work to get it changed. It'd be nice if they were reciprocal with ASCP and the other governing bodies--but this link will show you what we have to work with right now. Hope this helps! Cheryl Cheryl R. Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab, one great tech at a time. 281.852.9457 office 281.883.7704 cell 800.756.3309 fax and alternate phone From cbobrowi <@t> mcw.edu Tue Nov 6 11:51:29 2007 From: cbobrowi <@t> mcw.edu (Bobrowitz, Carol) Date: Tue Nov 6 11:51:37 2007 Subject: [Histonet] Gayle Callis Message-ID: <8F78639AC56F4143B267FE5F5A1B92C8F0AE74@guyton.phys.mcw.edu> Hi, I'm trying to e-mail Gayle Callis. It seems I have the wrong e-mail address. I also misplaced her phone number. Could anyone forward this information to me, please? Thank you, Carol Ann Bobrowitz Medical College of Wisconsin Department of Physiology cbobrowi@mcw.edu From vvancleave <@t> hendrickhealth.org Tue Nov 6 12:58:56 2007 From: vvancleave <@t> hendrickhealth.org (Vancleave, Vince) Date: Tue Nov 6 12:57:40 2007 Subject: [Histonet] non pathologist grossing versus processing Message-ID: <740F77F6594C20458BDC2A4A500D8D4150163C@EMAIL.hhs.hendrickhealth.org> Ok, So this is a subject that has been beat like a dead horse for a long time on histonet. The subject is, a histotech describing, measuring (color, description, measurments, etc) of G.I. and other non-complicated specimens (some skins included...inking). So, as a PA(ASCP) myself I spend much of my paid time with realy simple specimens like G.I. biopsies and skins. Basically, these are specimens that are boring and take up a lot of time cause there are so many of them....not to say it makes me not like my job....I still LOVE it. But, if I could delegate those to a histotech then I would have the time to devote more to higher complexity specimens like radical nephrectomies, mastectomies, pneumonectomies, etc.. We have some near by labs that our pathologists contract out to, of which also have gross, but those pathologists have been doing that gross. So, it would be great, in my mind to be able to deligate the low complexity specimens to technicians in both places and have myself grossing all the complex specimens so the pathologists don't have to hire another pathologist or PA and I would be expending what I get paid in a more effecient manner. In the CLIA regs it states that anyone doing gross exam on specimens needs to be a high complexity tester under CLIA 493.1489. It also mentions that there is a distinction between processing specimens (specimen preparation, what technicians do) and grossing specimens (describing color, weight, measurements, submitting sections). I notice on histonet that everybody always says that you have to be a 493.1489 tester to be able to do any type of gross. But, what if processing a specimen could include the description, measurement, inking, color, etc...of specimens that required no knowledge of anatomy and the entire specimen was submitted? According to the CAP inspection checklist, there is this exact destinction between two levels of macroscopic tissue complexity and are clearly defined. The two levels are defined as "1)Processing," and "2)Grossing." "Are all macroscopic tissue examinations performed by a pathologist or pathology resident, or under the supervision of a qualified pathologist? NOTE: Two levels of complexity of macroscopic tissue examination are defined, as follows: "1)Processing is defined as a tissue examination limited to description, inking and cutting of the specimen (if applicable), and submission of the entire specimen to histology. Tissue processing can be performed according to standardized protocols. Processing is generally limited to small specimens (skin ellipses, small biopsies, curettings, etc.) and does not require knowledge of anatomy." "2) Grossing (or gross examination) is defined as a tissue examination requiring a greater exercise of judgment and a knowledge of anatomy. Dissection of the specimen and selection of tissue samples for submission to histology are generally required. The specimen description is not necessarily standardized." Throughout this checklist it never requires one who processes tissues to be qualified as a high complexity tester, or 493.1489 individual. You must, however, include a lot of documentation about the nature of supervision (direct or indirect, by a pathologist) and the specific types of specimens that they are limited to, and exactly how they are to process them. My question is, why don't we here more about this? Why can't I find it in the CLIA regs, but I can in the CAP checklist? And, is anybody out there even aware of this disctinction? Well, it made my day...and hopefully it will be a help to others out there! Vince Van Cleave, BS, PA(ASCP), HTL, HT Laboratory Manager and Pathologist Assistant (325)670-6528 Clinical Pathology Associates Abilene, TX PRIVILEGED AND CONFIDENTIAL NOTICE: The information contained in this e-mail may be confidential and/or privileged. This e-mail is intended to be reviewed initially by only the individual named above. If the reader of this e-mail is not the intended recipient or a representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail, or the information contained herein, is prohibited. If you have received this e-mail in error, notify the above sender, send the message back and then delete it. Thank you for your assistance. Hendrick Health System 1900 Pine St. Abilene, Texas 79601 - 325.670.2000 79601 - 325.670.2000 From JWEEMS <@t> sjha.org Tue Nov 6 13:09:04 2007 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Tue Nov 6 13:09:21 2007 Subject: [Histonet] non pathologist grossing versus processing In-Reply-To: <740F77F6594C20458BDC2A4A500D8D4150163C@EMAIL.hhs.hendrickhealth.org> Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD32048F4165@sjhaexc02.sjha.org> Bottom line - you must be high complexity for CLIA and you must have distinct procedures for CAP. Mine said "Biopsy Grossing", so at inspection I had to change it to processing. Which for us means the tissue processor. So it's all a mess, in my most humble 2 cents worth again! j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Vancleave, Vince Sent: Tuesday, November 06, 2007 1:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] non pathologist grossing versus processing Ok, So this is a subject that has been beat like a dead horse for a long time on histonet. The subject is, a histotech describing, measuring (color, description, measurments, etc) of G.I. and other non-complicated specimens (some skins included...inking). So, as a PA(ASCP) myself I spend much of my paid time with realy simple specimens like G.I. biopsies and skins. Basically, these are specimens that are boring and take up a lot of time cause there are so many of them....not to say it makes me not like my job....I still LOVE it. But, if I could delegate those to a histotech then I would have the time to devote more to higher complexity specimens like radical nephrectomies, mastectomies, pneumonectomies, etc.. We have some near by labs that our pathologists contract out to, of which also have gross, but those pathologists have been doing that gross. So, it would be great, in my mind to be able to deligate the low complexity specimens to technicians in both places and have myself grossing all the complex specimens so the pathologists don't have to hire another pathologist or PA and I would be expending what I get paid in a more effecient manner. In the CLIA regs it states that anyone doing gross exam on specimens needs to be a high complexity tester under CLIA 493.1489. It also mentions that there is a distinction between processing specimens (specimen preparation, what technicians do) and grossing specimens (describing color, weight, measurements, submitting sections). I notice on histonet that everybody always says that you have to be a 493.1489 tester to be able to do any type of gross. But, what if processing a specimen could include the description, measurement, inking, color, etc...of specimens that required no knowledge of anatomy and the entire specimen was submitted? According to the CAP inspection checklist, there is this exact destinction between two levels of macroscopic tissue complexity and are clearly defined. The two levels are defined as "1)Processing," and "2)Grossing." "Are all macroscopic tissue examinations performed by a pathologist or pathology resident, or under the supervision of a qualified pathologist? NOTE: Two levels of complexity of macroscopic tissue examination are defined, as follows: "1)Processing is defined as a tissue examination limited to description, inking and cutting of the specimen (if applicable), and submission of the entire specimen to histology. Tissue processing can be performed according to standardized protocols. Processing is generally limited to small specimens (skin ellipses, small biopsies, curettings, etc.) and does not require knowledge of anatomy." "2) Grossing (or gross examination) is defined as a tissue examination requiring a greater exercise of judgment and a knowledge of anatomy. Dissection of the specimen and selection of tissue samples for submission to histology are generally required. The specimen description is not necessarily standardized." Throughout this checklist it never requires one who processes tissues to be qualified as a high complexity tester, or 493.1489 individual. You must, however, include a lot of documentation about the nature of supervision (direct or indirect, by a pathologist) and the specific types of specimens that they are limited to, and exactly how they are to process them. My question is, why don't we here more about this? Why can't I find it in the CLIA regs, but I can in the CAP checklist? And, is anybody out there even aware of this disctinction? Well, it made my day...and hopefully it will be a help to others out there! Vince Van Cleave, BS, PA(ASCP), HTL, HT Laboratory Manager and Pathologist Assistant (325)670-6528 Clinical Pathology Associates Abilene, TX PRIVILEGED AND CONFIDENTIAL NOTICE: The information contained in this e-mail may be confidential and/or privileged. This e-mail is intended to be reviewed initially by only the individual named above. If the reader of this e-mail is not the intended recipient or a representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail, or the information contained herein, is prohibited. If you have received this e-mail in error, notify the above sender, send the message back and then delete it. Thank you for your assistance. Hendrick Health System 1900 Pine St. Abilene, Texas 79601 - 325.670.2000 79601 - 325.670.2000 _______________________________________________ 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 Joseph's Health System, Inc. From dez <@t> u.washington.edu Tue Nov 6 13:24:21 2007 From: dez <@t> u.washington.edu (J. Fernandez) Date: Tue Nov 6 13:24:30 2007 Subject: [Histonet] non pathologist grossing versus processing In-Reply-To: <740F77F6594C20458BDC2A4A500D8D4150163C@EMAIL.hhs.hendrickhealth.org> References: <740F77F6594C20458BDC2A4A500D8D4150163C@EMAIL.hhs.hendrickhealth.org> Message-ID: joeltao64@aol.com On Tue, 6 Nov 2007, Vancleave, Vince wrote: > Ok, > So this is a subject that has been beat like a dead horse for a long time on histonet. The subject is, a histotech describing, measuring (color, description, measurments, etc) of G.I. and other non-complicated specimens (some skins included...inking). > > So, as a PA(ASCP) myself I spend much of my paid time with realy simple specimens like G.I. biopsies and skins. Basically, these are specimens that are boring and take up a lot of time cause there are so many of them....not to say it makes me not like my job....I still LOVE it. But, if I could delegate those to a histotech then I would have the time to devote more to higher complexity specimens like radical nephrectomies, mastectomies, pneumonectomies, etc.. We have some near by labs that our pathologists contract out to, of which also have gross, but those pathologists have been doing that gross. So, it would be great, in my mind to be able to deligate the low complexity specimens to technicians in both places and have myself grossing all the complex specimens so the pathologists don't have to hire another pathologist or PA and I would be expending what I get paid in a more effecient manner. > > In the CLIA regs it states that anyone doing gross exam on specimens needs to be a high complexity tester under CLIA 493.1489. It also mentions that there is a distinction between processing specimens (specimen preparation, what technicians do) and grossing specimens (describing color, weight, measurements, submitting sections). I notice on histonet that everybody always says that you have to be a 493.1489 tester to be able to do any type of gross. But, what if processing a specimen could include the description, measurement, inking, color, etc...of specimens that required no knowledge of anatomy and the entire specimen was submitted? According to the CAP inspection checklist, there is this exact destinction between two levels of macroscopic tissue complexity and are clearly defined. The two levels are defined as "1)Processing," and "2)Grossing." > > > "Are all macroscopic tissue examinations performed by a pathologist or pathology resident, or under the supervision of a qualified pathologist? > > NOTE: Two levels of complexity of macroscopic tissue examination are defined, as follows: > > > "1)Processing is defined as a tissue examination limited to description, inking and cutting of the specimen (if applicable), and submission of the entire specimen to histology. Tissue processing can be performed according to standardized protocols. Processing is generally limited to small specimens (skin ellipses, small biopsies, curettings, etc.) and does not require knowledge of anatomy." > > "2) Grossing (or gross examination) is defined as a tissue examination requiring a greater exercise of judgment and a knowledge of anatomy. Dissection of the specimen and selection of tissue samples for submission to histology are generally required. The specimen description is not necessarily standardized." > > > Throughout this checklist it never requires one who processes tissues to be qualified as a high complexity tester, or 493.1489 individual. You must, however, include a lot of documentation about the nature of supervision (direct or indirect, by a pathologist) and the specific types of specimens that they are limited to, and exactly how they are to process them. > > My question is, why don't we here more about this? Why can't I find it in the CLIA regs, but I can in the CAP checklist? And, is anybody out there even aware of this disctinction? Well, it made my day...and hopefully it will be a help to others out there! > > > Vince Van Cleave, BS, PA(ASCP), HTL, HT > Laboratory Manager and Pathologist Assistant > (325)670-6528 > Clinical Pathology Associates > Abilene, TX > > PRIVILEGED AND CONFIDENTIAL NOTICE: > The information contained in this e-mail may be confidential and/or privileged. This e-mail is intended to be reviewed initially by only the individual named above. If the reader of this e-mail is not the intended recipient or a representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail, or the information contained herein, is prohibited. If you have received this e-mail in error, notify the above sender, send the message back and then delete it. Thank you for your assistance. > > Hendrick Health System 1900 Pine St. Abilene, Texas 79601 - 325.670.2000 79601 - 325.670.2000 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Melissa.Gonzalez <@t> cellgenesys.com Tue Nov 6 13:37:24 2007 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Tue Nov 6 13:37:33 2007 Subject: [Histonet] RE: eosinophil peroxidase In-Reply-To: <6c784e$u5ac@mail.cellgenesys.com> References: <6c784e$u5ac@mail.cellgenesys.com> Message-ID: <2884B897182A1D438C7BA24B9A8F94A213A47F@hqsvr01mail.cgi.com> Hi Michelle, I have also only seen one publication for EPO IHC but it was also on frozens with mouse tissue. Have you tried MBP, ECP (via EG1 & EG2), EDN, or eotaxin on human FFPE's with any success? Melissa Melissa A. Gonz?lez Edick R&D, Cell Genesys Inc. 500 Forbes Blvd South San Francisco, CA 94080 p(650) 266-3168 f (650) 266-3080 ------------------------------ Message: 2 Date: Tue, 6 Nov 2007 08:57:25 -0600 From: "Griffin-Reyes, Michelle A" Subject: [Histonet] eosinophil peroxidase (EPO) To: Does anyone have any experience using eosinophil peroxidase (EPO) to stain FFPE tissues (I am doing other staining methods as well)? I have searched and can find EPO usage for western blot and flow and one publication using frozen tissues but nothing for FFPE tissues. Any suggestions would be greatly appreciated. Thanks in advance!! Michelle A. Griffin-Reyes Comparative Pathology Laboratory University of Iowa Hospitals and Clinics 319-384-4620 From cbobrowi <@t> mcw.edu Tue Nov 6 14:10:56 2007 From: cbobrowi <@t> mcw.edu (Bobrowitz, Carol) Date: Tue Nov 6 14:11:05 2007 Subject: [Histonet] primary antibodies specific for rat tissue Message-ID: <8F78639AC56F4143B267FE5F5A1B92C8F0AE75@guyton.phys.mcw.edu> Hi, I'm so jealous that Gayle has retired. I wish her much happiness. I'm looking for sources/vendors that carry primary antibodies specific for rat tissue. I'm finding many companies have merged and I need some direction where to look. Any information will be greatly appreciated. Thank you, Carol Ann Bobrowitz Medical College of Wisconsin Department of Physiology cbobrowi@mcw.edu From portera <@t> msu.edu Tue Nov 6 14:50:26 2007 From: portera <@t> msu.edu (Amy Porter) Date: Tue Nov 6 14:52:40 2007 Subject: [Histonet] primary antibodies specific for rat tissue References: <8F78639AC56F4143B267FE5F5A1B92C8F0AE75@guyton.phys.mcw.edu> Message-ID: <002601c820b6$a8bd63e0$8e7a0923@histolab> Try AbD Serotec, BD Pharmingen, or Serotec. Amy S. Porter, HT (ASCP) QIHC Investigative HistoPathology Laboratory - Supervisor 2201 Biomedical Physical Sciences Bldg. Rm #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 Fax: (517) 432-1368 Email: portera@msu.edu Web: www.humanpathology.msu.edu ----- Original Message ----- From: "Bobrowitz, Carol" To: Sent: Tuesday, November 06, 2007 3:10 PM Subject: [Histonet] primary antibodies specific for rat tissue Hi, I'm so jealous that Gayle has retired. I wish her much happiness. I'm looking for sources/vendors that carry primary antibodies specific for rat tissue. I'm finding many companies have merged and I need some direction where to look. Any information will be greatly appreciated. Thank you, Carol Ann Bobrowitz Medical College of Wisconsin Department of Physiology cbobrowi@mcw.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Tue Nov 6 14:55:07 2007 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Tue Nov 6 14:55:22 2007 Subject: [Histonet] primary antibodies specific for rat tissue In-Reply-To: <8F78639AC56F4143B267FE5F5A1B92C8F0AE75@guyton.phys.mcw.edu> Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD32048F416F@sjhaexc02.sjha.org> Who gave her permission?! I hope she hasn't left us for good! Gayle if you you're there, best wishes! Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bobrowitz, Carol Sent: Tuesday, November 06, 2007 3:11 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] primary antibodies specific for rat tissue Hi, I'm so jealous that Gayle has retired. I wish her much happiness. I'm looking for sources/vendors that carry primary antibodies specific for rat tissue. I'm finding many companies have merged and I need some direction where to look. Any information will be greatly appreciated. Thank you, Carol Ann Bobrowitz Medical College of Wisconsin Department of Physiology cbobrowi@mcw.edu _______________________________________________ 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 Joseph's Health System, Inc. From Linke_Noelle <@t> Allergan.com Tue Nov 6 15:47:58 2007 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Tue Nov 6 15:48:55 2007 Subject: [Histonet] primary antibodies specific for rat tissue In-Reply-To: <8F78639AC56F4143B267FE5F5A1B92C8F0AE75@guyton.phys.mcw.edu> Message-ID: <5C3DA4BE34AA0641BAA10A7C1478B60502EC3A@IRMAIL132.irvine.allergan.com> LabVision (Thermo Fisher) is my favorite.... No?lle Linke, MS, 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 Bobrowitz, Carol Sent: Tuesday, November 06, 2007 12:11 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] primary antibodies specific for rat tissue Hi, I'm so jealous that Gayle has retired. I wish her much happiness. I'm looking for sources/vendors that carry primary antibodies specific for rat tissue. I'm finding many companies have merged and I need some direction where to look. Any information will be greatly appreciated. Thank you, Carol Ann Bobrowitz Medical College of Wisconsin Department of Physiology cbobrowi@mcw.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Charles.Embrey <@t> carle.com Tue Nov 6 16:06:53 2007 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Tue Nov 6 16:08:10 2007 Subject: [Histonet] non pathologist grossing versus processing In-Reply-To: <740F77F6594C20458BDC2A4A500D8D4150163C@EMAIL.hhs.hendrickhealth.org> Message-ID: <44780C571F28624DBB446DE55C4D733A1FE566@EXCHANGEBE1.carle.com> The thing you have to remember, Vince, is that CLIA and CAP are two different, completely separate entities. CLIA is government controlled and CAP is private. Whereas CAP recently decided to separate grossing into two different complexities, CLIA has not. Bottom line...if you are CLIA licensed you MUST follow the CLIA standard. If not, then you can do whatever you like. Charles Embrey Jr., PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vancleave, Vince Sent: Tuesday, November 06, 2007 12:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] non pathologist grossing versus processing Ok, So this is a subject that has been beat like a dead horse for a long time on histonet. The subject is, a histotech describing, measuring (color, description, measurments, etc) of G.I. and other non-complicated specimens (some skins included...inking). So, as a PA(ASCP) myself I spend much of my paid time with realy simple specimens like G.I. biopsies and skins. Basically, these are specimens that are boring and take up a lot of time cause there are so many of them....not to say it makes me not like my job....I still LOVE it. But, if I could delegate those to a histotech then I would have the time to devote more to higher complexity specimens like radical nephrectomies, mastectomies, pneumonectomies, etc.. We have some near by labs that our pathologists contract out to, of which also have gross, but those pathologists have been doing that gross. So, it would be great, in my mind to be able to deligate the low complexity specimens to technicians in both places and have myself grossing all the complex specimens so the pathologists don't have to hire another pathologist or PA and I would be expending what I get paid in a more effecient manner. In the CLIA regs it states that anyone doing gross exam on specimens needs to be a high complexity tester under CLIA 493.1489. It also mentions that there is a distinction between processing specimens (specimen preparation, what technicians do) and grossing specimens (describing color, weight, measurements, submitting sections). I notice on histonet that everybody always says that you have to be a 493.1489 tester to be able to do any type of gross. But, what if processing a specimen could include the description, measurement, inking, color, etc...of specimens that required no knowledge of anatomy and the entire specimen was submitted? According to the CAP inspection checklist, there is this exact destinction between two levels of macroscopic tissue complexity and are clearly defined. The two levels are defined as "1)Processing," and "2)Grossing." "Are all macroscopic tissue examinations performed by a pathologist or pathology resident, or under the supervision of a qualified pathologist? NOTE: Two levels of complexity of macroscopic tissue examination are defined, as follows: "1)Processing is defined as a tissue examination limited to description, inking and cutting of the specimen (if applicable), and submission of the entire specimen to histology. Tissue processing can be performed according to standardized protocols. Processing is generally limited to small specimens (skin ellipses, small biopsies, curettings, etc.) and does not require knowledge of anatomy." "2) Grossing (or gross examination) is defined as a tissue examination requiring a greater exercise of judgment and a knowledge of anatomy. Dissection of the specimen and selection of tissue samples for submission to histology are generally required. The specimen description is not necessarily standardized." Throughout this checklist it never requires one who processes tissues to be qualified as a high complexity tester, or 493.1489 individual. You must, however, include a lot of documentation about the nature of supervision (direct or indirect, by a pathologist) and the specific types of specimens that they are limited to, and exactly how they are to process them. My question is, why don't we here more about this? Why can't I find it in the CLIA regs, but I can in the CAP checklist? And, is anybody out there even aware of this disctinction? Well, it made my day...and hopefully it will be a help to others out there! Vince Van Cleave, BS, PA(ASCP), HTL, HT Laboratory Manager and Pathologist Assistant (325)670-6528 Clinical Pathology Associates Abilene, TX PRIVILEGED AND CONFIDENTIAL NOTICE: The information contained in this e-mail may be confidential and/or privileged. This e-mail is intended to be reviewed initially by only the individual named above. If the reader of this e-mail is not the intended recipient or a representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail, or the information contained herein, is prohibited. If you have received this e-mail in error, notify the above sender, send the message back and then delete it. Thank you for your assistance. Hendrick Health System 1900 Pine St. Abilene, Texas 79601 - 325.670.2000 79601 - 325.670.2000 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjchtascp <@t> yahoo.com Tue Nov 6 17:49:24 2007 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Tue Nov 6 17:49:31 2007 Subject: [Histonet] Florida Contract work Message-ID: <932048.10178.qm@web38211.mail.mud.yahoo.com> How does an HT get a FL license in order to contract in Florida. I'm mainly interested in Jacksonville. Steve __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From LuckG <@t> empirehealth.org Tue Nov 6 17:59:02 2007 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Tue Nov 6 17:59:11 2007 Subject: [Histonet] non pathologist grossing versus processing In-Reply-To: <44780C571F28624DBB446DE55C4D733A1FE566@EXCHANGEBE1.carle.com> References: <740F77F6594C20458BDC2A4A500D8D4150163C@EMAIL.hhs.hendrickhealth.org> <44780C571F28624DBB446DE55C4D733A1FE566@EXCHANGEBE1.carle.com> Message-ID: <6BB8BC4519AAB844B174FC739A679BBCCEFF08@IRMEXCH01.irm.inhs.org> Charles, et.al. , Which standard do you follow if your lab is both CLIA licensed/regulated and CAP accredited? It would seem to me that the deemed status would fall to the higher/more stringent standard (i.e. in this case CLIA). Thanks, Greg Greg Luck, B.S., HT(ASCP) Anatomic Pathology Supervisor Deaconess Med Cntr 800 W. 5th Ave Spokane, WA 99204 Offc 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org www.deaconessmc.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Tuesday, November 06, 2007 2:07 PM To: Vancleave, Vince; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] non pathologist grossing versus processing The thing you have to remember, Vince, is that CLIA and CAP are two different, completely separate entities. CLIA is government controlled and CAP is private. Whereas CAP recently decided to separate grossing into two different complexities, CLIA has not. Bottom line...if you are CLIA licensed you MUST follow the CLIA standard. If not, then you can do whatever you like. Charles Embrey Jr., PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vancleave, Vince Sent: Tuesday, November 06, 2007 12:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] non pathologist grossing versus processing Ok, So this is a subject that has been beat like a dead horse for a long time on histonet. The subject is, a histotech describing, measuring (color, description, measurments, etc) of G.I. and other non-complicated specimens (some skins included...inking). So, as a PA(ASCP) myself I spend much of my paid time with realy simple specimens like G.I. biopsies and skins. Basically, these are specimens that are boring and take up a lot of time cause there are so many of them....not to say it makes me not like my job....I still LOVE it. But, if I could delegate those to a histotech then I would have the time to devote more to higher complexity specimens like radical nephrectomies, mastectomies, pneumonectomies, etc.. We have some near by labs that our pathologists contract out to, of which also have gross, but those pathologists have been doing that gross. So, it would be great, in my mind to be able to deligate the low complexity specimens to technicians in both places and have myself grossing all the complex specimens so the pathologists don't have to hire another pathologist or PA and I would be expending what I get paid in a more effecient manner. In the CLIA regs it states that anyone doing gross exam on specimens needs to be a high complexity tester under CLIA 493.1489. It also mentions that there is a distinction between processing specimens (specimen preparation, what technicians do) and grossing specimens (describing color, weight, measurements, submitting sections). I notice on histonet that everybody always says that you have to be a 493.1489 tester to be able to do any type of gross. But, what if processing a specimen could include the description, measurement, inking, color, etc...of specimens that required no knowledge of anatomy and the entire specimen was submitted? According to the CAP inspection checklist, there is this exact destinction between two levels of macroscopic tissue complexity and are clearly defined. The two levels are defined as "1)Processing," and "2)Grossing." "Are all macroscopic tissue examinations performed by a pathologist or pathology resident, or under the supervision of a qualified pathologist? NOTE: Two levels of complexity of macroscopic tissue examination are defined, as follows: "1)Processing is defined as a tissue examination limited to description, inking and cutting of the specimen (if applicable), and submission of the entire specimen to histology. Tissue processing can be performed according to standardized protocols. Processing is generally limited to small specimens (skin ellipses, small biopsies, curettings, etc.) and does not require knowledge of anatomy." "2) Grossing (or gross examination) is defined as a tissue examination requiring a greater exercise of judgment and a knowledge of anatomy. Dissection of the specimen and selection of tissue samples for submission to histology are generally required. The specimen description is not necessarily standardized." Throughout this checklist it never requires one who processes tissues to be qualified as a high complexity tester, or 493.1489 individual. You must, however, include a lot of documentation about the nature of supervision (direct or indirect, by a pathologist) and the specific types of specimens that they are limited to, and exactly how they are to process them. My question is, why don't we here more about this? Why can't I find it in the CLIA regs, but I can in the CAP checklist? And, is anybody out there even aware of this disctinction? Well, it made my day...and hopefully it will be a help to others out there! Vince Van Cleave, BS, PA(ASCP), HTL, HT Laboratory Manager and Pathologist Assistant (325)670-6528 Clinical Pathology Associates Abilene, TX PRIVILEGED AND CONFIDENTIAL NOTICE: The information contained in this e-mail may be confidential and/or privileged. This e-mail is intended to be reviewed initially by only the individual named above. If the reader of this e-mail is not the intended recipient or a representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail, or the information contained herein, is prohibited. If you have received this e-mail in error, notify the above sender, send the message back and then delete it. Thank you for your assistance. Hendrick Health System 1900 Pine St. Abilene, Texas 79601 - 325.670.2000 79601 - 325.670.2000 _______________________________________________ Histonet mailing list Histonet@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 khenken <@t> ufl.edu Wed Nov 7 08:43:32 2007 From: khenken <@t> ufl.edu (HENKEN,KIRSTEN RUTH) Date: Wed Nov 7 08:43:43 2007 Subject: [Histonet] viability stain for 450 micron brain slices Message-ID: <464094433.180371194446612705.JavaMail.osg@osgjas03.cns.ufl.edu> I am looking for a staining technique to test the viability of slices of the rat brain after some force-displacement tests on the tissue. As I am an engineer and no chemist I do not know what kind of stain would be appropriate. We keep the slices from the cerebral cortex and the corpus callosumalive alive in a tissue chamber with ACSF that is bubbled with 95% O2/ 5% CO2. The slices have a thickness of 450 micron and there is a fluorescent confocal microscope available. As this microscope is away from the testing site and we do not have the equipment that can keep the tissue alive during imaging, it would be very convenient to fix the tissue after staining. Does anyone of you have experience with a staining technique that is suitable for my experiment? Best, Kirsten Henken From Charles.Embrey <@t> carle.com Wed Nov 7 08:54:44 2007 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Wed Nov 7 08:56:02 2007 Subject: [Histonet] non pathologist grossing versus processing In-Reply-To: <6BB8BC4519AAB844B174FC739A679BBCCEFF08@IRMEXCH01.irm.inhs.org> Message-ID: <44780C571F28624DBB446DE55C4D733A1FE568@EXCHANGEBE1.carle.com> Exactly. If you fall under both standards the more stringent must be followed. You can't get in trouble for going above the standard just for being below it. By meeting the CLIA standard you pass both your CAP and CLIA inspections. Chuck -----Original Message----- From: Luck, Greg D. [mailto:LuckG@empirehealth.org] Sent: Tuesday, November 06, 2007 5:59 PM To: Charles.Embrey; Vancleave, Vince; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] non pathologist grossing versus processing Charles, et.al. , Which standard do you follow if your lab is both CLIA licensed/regulated and CAP accredited? It would seem to me that the deemed status would fall to the higher/more stringent standard (i.e. in this case CLIA). Thanks, Greg Greg Luck, B.S., HT(ASCP) Anatomic Pathology Supervisor Deaconess Med Cntr 800 W. 5th Ave Spokane, WA 99204 Offc 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org www.deaconessmc.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Tuesday, November 06, 2007 2:07 PM To: Vancleave, Vince; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] non pathologist grossing versus processing The thing you have to remember, Vince, is that CLIA and CAP are two different, completely separate entities. CLIA is government controlled and CAP is private. Whereas CAP recently decided to separate grossing into two different complexities, CLIA has not. Bottom line...if you are CLIA licensed you MUST follow the CLIA standard. If not, then you can do whatever you like. Charles Embrey Jr., PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vancleave, Vince Sent: Tuesday, November 06, 2007 12:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] non pathologist grossing versus processing Ok, So this is a subject that has been beat like a dead horse for a long time on histonet. The subject is, a histotech describing, measuring (color, description, measurments, etc) of G.I. and other non-complicated specimens (some skins included...inking). So, as a PA(ASCP) myself I spend much of my paid time with realy simple specimens like G.I. biopsies and skins. Basically, these are specimens that are boring and take up a lot of time cause there are so many of them....not to say it makes me not like my job....I still LOVE it. But, if I could delegate those to a histotech then I would have the time to devote more to higher complexity specimens like radical nephrectomies, mastectomies, pneumonectomies, etc.. We have some near by labs that our pathologists contract out to, of which also have gross, but those pathologists have been doing that gross. So, it would be great, in my mind to be able to deligate the low complexity specimens to technicians in both places and have myself grossing all the complex specimens so the pathologists don't have to hire another pathologist or PA and I would be expending what I get paid in a more effecient manner. In the CLIA regs it states that anyone doing gross exam on specimens needs to be a high complexity tester under CLIA 493.1489. It also mentions that there is a distinction between processing specimens (specimen preparation, what technicians do) and grossing specimens (describing color, weight, measurements, submitting sections). I notice on histonet that everybody always says that you have to be a 493.1489 tester to be able to do any type of gross. But, what if processing a specimen could include the description, measurement, inking, color, etc...of specimens that required no knowledge of anatomy and the entire specimen was submitted? According to the CAP inspection checklist, there is this exact destinction between two levels of macroscopic tissue complexity and are clearly defined. The two levels are defined as "1)Processing," and "2)Grossing." "Are all macroscopic tissue examinations performed by a pathologist or pathology resident, or under the supervision of a qualified pathologist? NOTE: Two levels of complexity of macroscopic tissue examination are defined, as follows: "1)Processing is defined as a tissue examination limited to description, inking and cutting of the specimen (if applicable), and submission of the entire specimen to histology. Tissue processing can be performed according to standardized protocols. Processing is generally limited to small specimens (skin ellipses, small biopsies, curettings, etc.) and does not require knowledge of anatomy." "2) Grossing (or gross examination) is defined as a tissue examination requiring a greater exercise of judgment and a knowledge of anatomy. Dissection of the specimen and selection of tissue samples for submission to histology are generally required. The specimen description is not necessarily standardized." Throughout this checklist it never requires one who processes tissues to be qualified as a high complexity tester, or 493.1489 individual. You must, however, include a lot of documentation about the nature of supervision (direct or indirect, by a pathologist) and the specific types of specimens that they are limited to, and exactly how they are to process them. My question is, why don't we here more about this? Why can't I find it in the CLIA regs, but I can in the CAP checklist? And, is anybody out there even aware of this disctinction? Well, it made my day...and hopefully it will be a help to others out there! Vince Van Cleave, BS, PA(ASCP), HTL, HT Laboratory Manager and Pathologist Assistant (325)670-6528 Clinical Pathology Associates Abilene, TX PRIVILEGED AND CONFIDENTIAL NOTICE: The information contained in this e-mail may be confidential and/or privileged. This e-mail is intended to be reviewed initially by only the individual named above. If the reader of this e-mail is not the intended recipient or a representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail, or the information contained herein, is prohibited. If you have received this e-mail in error, notify the above sender, send the message back and then delete it. Thank you for your assistance. Hendrick Health System 1900 Pine St. Abilene, Texas 79601 - 325.670.2000 79601 - 325.670.2000 _______________________________________________ Histonet mailing list Histonet@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 laurie.colbert <@t> huntingtonhospital.com Wed Nov 7 09:51:55 2007 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Wed Nov 7 09:52:00 2007 Subject: [Histonet] State Requirements for Keeping Records Message-ID: <57BE698966D5C54EAE8612E8941D768301EA66ED@EXCHANGE3.huntingtonhospital.com> Does anyone know what the state requirement for California is for keeping records, such as log sheets, and where I can find documentation stating these requirements? Laurie Colbert Huntington Hospital From TillRenee <@t> uams.edu Wed Nov 7 11:08:12 2007 From: TillRenee <@t> uams.edu (Till, Renee) Date: Wed Nov 7 11:08:41 2007 Subject: [Histonet] Rocking/shaking slides Message-ID: <11F927674DEBDC43B960809A7403C5D206529641@MAILPED.ad.uams.edu> Does anyone rock or shake their slides during incubation steps? I recently had a product e-mail given to me for a slide container intended for this use. The theory being the same as used during western blots; that it would reduce the antibody concentration needed for good staining and that the antibody solution would be reusable. I have protocols that call for washing steps on a shaker, but I had never thought of how it might affect the actual antibody incubation nor come across a protocol that calls for it. Renee' Till, HT (ASCP) Research Assistant Arkansas Children's Nutrition Center 1212 Marshall St./N2021 Little Rock, AR 72202 Lab (501)364-8504 Office Fax (501)364-3161 Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From katia.catunda <@t> cipax.com.br Wed Nov 7 12:20:36 2007 From: katia.catunda <@t> cipax.com.br (=?iso-8859-1?Q?Ms._K=E1tia_Cristina_Catunda?=) Date: Wed Nov 7 11:20:16 2007 Subject: [Histonet] Templates for grossing reports In-Reply-To: <44780C571F28624DBB446DE55C4D733A1FE568@EXCHANGEBE1.carle.com> Message-ID: Hi there again, Hope I can make myself clear with the following question: Does anybody uses preformatted templates for gross description on the reports? Have you developed them or bought? We actually use a combination of many codes, each code for each one of the characteristic of the specimen. Each code is written on a paper and sent to another department so the codes may be inserted on the computer, and the computer automatically translate each code to a phrase. Our goal is to create one unique code for each kind of specimen received. Tks a lot! ? Ms. K?tia Catunda Produ??o +55 12 3203-0612 (direto) +55 12 3203-0633 (PABX) www.cipax.com.br katia.catunda@cipax.com.br From Rcartun <@t> harthosp.org Wed Nov 7 11:31:02 2007 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Wed Nov 7 11:31:20 2007 Subject: [Histonet] Fli-1 antibody Message-ID: <4731B0060200007700008F8C@gwmail4.harthosp.org> Which antibody are people using for the immunohistochemical detection of Fli-1 in formalin-fixed tissues? We are currently using a rabbit polyclonal from Santa Cruz labeled "RUO". 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 rjbuesa <@t> yahoo.com Wed Nov 7 11:58:45 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 7 11:58:49 2007 Subject: [Histonet] Rocking/shaking slides In-Reply-To: <11F927674DEBDC43B960809A7403C5D206529641@MAILPED.ad.uams.edu> Message-ID: <303300.99903.qm@web61211.mail.yahoo.com> Totally unnecessary, just a good sales pitch worth disregarding. Ren? J. "Till, Renee" wrote: Does anyone rock or shake their slides during incubation steps? I recently had a product e-mail given to me for a slide container intended for this use. The theory being the same as used during western blots; that it would reduce the antibody concentration needed for good staining and that the antibody solution would be reusable. I have protocols that call for washing steps on a shaker, but I had never thought of how it might affect the actual antibody incubation nor come across a protocol that calls for it. Renee' Till, HT (ASCP) Research Assistant Arkansas Children's Nutrition Center 1212 Marshall St./N2021 Little Rock, AR 72202 Lab (501)364-8504 Office Fax (501)364-3161 Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From burch007 <@t> mc.duke.edu Wed Nov 7 12:05:48 2007 From: burch007 <@t> mc.duke.edu (James L Burchette) Date: Wed Nov 7 12:04:16 2007 Subject: [Histonet] Fli-1 antibody In-Reply-To: <4731B0060200007700008F8C@gwmail4.harthosp.org> Message-ID: Richard: We are using a rabbit polyclonal Fli-1 from Thermo Scientific. It's RUO classified. Jim "Richard Cartun" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/07/2007 12:31 PM To "Histonet" cc Subject [Histonet] Fli-1 antibody Which antibody are people using for the immunohistochemical detection of Fli-1 in formalin-fixed tissues? We are currently using a rabbit polyclonal from Santa Cruz labeled "RUO". 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 arnie.jimenez <@t> vel-lab.com Wed Nov 7 12:08:19 2007 From: arnie.jimenez <@t> vel-lab.com (arnie.jimenez@vel-lab.com) Date: Wed Nov 7 12:10:22 2007 Subject: [Histonet] Re: Away from Office Message-ID: <20071107180819.10376.qmail@plesk5.hostgator.com> Thank you for contacting Vel-Lab Research. We will be out of the lab starting Nov. 7th and will return on Nov. 12th. We will check e-mail regularly but will be unable to respond. Please do not mail any packages to our lab until Nov. 12th. Regards, Arnie Jimenez Vel-Lab Research From algranth <@t> u.arizona.edu Wed Nov 7 12:43:10 2007 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Wed Nov 7 12:48:20 2007 Subject: [Histonet] Cape Fear Hospital Message-ID: <6.2.3.4.1.20071107112809.01fffa58@algranth.inbox.email.arizona.edu> Is there a histotech on this listserver from Cape Fear Hospital in Wilmington NC? If so - I have a student who will graduate from our Histology program and be moving there next month. She is eligible for the ASCP HT Registry and will be looking for a job when she gets there. We were wondering if you still had an opening in the histology lab or knew of any histology jobs in the area. Thanks! Andi Grantham ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From jdhisto <@t> yahoo.com Wed Nov 7 13:37:47 2007 From: jdhisto <@t> yahoo.com (Jonathan De La Rosa) Date: Wed Nov 7 13:37:49 2007 Subject: [Histonet] Cape Fear Hospital Message-ID: <444026.82073.qm@web30307.mail.mud.yahoo.com> The Hospital in Wilmington, NC is "New Hanover Regional Medical Center", part of the New Hanover Health Network (NHHN). I'm pretty sure they might have a position. You can contact the laboratory at 910.343.2718. http://www.nhhn.org/ There's another lab down the street form the hospital that hires Histotechs. I think the name is "WPA (Wilmington Pathology Associates)". http://www.wilmingtonpathology.com/. The Dr's, Manager's, techs, laboratory set up and city are great. Good luck; tell them Jonathan sent ya. Jonathan De La Rosa From Jessica.Vacca <@t> HCAhealthcare.com Wed Nov 7 13:39:19 2007 From: Jessica.Vacca <@t> HCAhealthcare.com (Vacca Jessica) Date: Wed Nov 7 13:39:45 2007 Subject: [Histonet] Autopsy table Message-ID: <41E16A15CE78374EA45B57E0F94339B802FD78DE@ORLEV01.hca.corpad.net> We will no longer be doing our autopsies in-house. Does anyone know of anyone or anywhere that might be interested in purchasing this used piece of equipment? It looks like a Jewett autopsy table model "D". If anyone is interested please contact me. Thanks Jessica Vacca Histology Supervisor Brandon Regional Hospital 119 Oakfield Dr. Brandon Fl 33511 (813) 571-5193 or (813) 681-5551 ext 2454 Jessica.Vacca@hcahealthcare.com From LBlack <@t> carilion.com Wed Nov 7 13:59:29 2007 From: LBlack <@t> carilion.com (Lisa Black) Date: Wed Nov 7 13:59:56 2007 Subject: [Histonet] Histology position available Message-ID: <4731D2D1020000E80000B699@chs-gw-gwia2.carilion.com> We have a full time Histology position open at Carilion Labs in Roanoke, VA. If anyone is interested, please go to www.carilion.com and look at the job postings. If you would like more information, feel free to give me a call. Lisa Black Histology Manager Carilion Labs Roanoke, VA 540-985-4082 From vvancleave <@t> hendrickhealth.org Wed Nov 7 14:39:28 2007 From: vvancleave <@t> hendrickhealth.org (Vancleave, Vince) Date: Wed Nov 7 14:38:18 2007 Subject: [Histonet] Still not sure about this low complexity or not Message-ID: <740F77F6594C20458BDC2A4A500D8D4150163F@EMAIL.hhs.hendrickhealth.org> "The thing you have to remember, Vince, is that CLIA and CAP are two different, completely separate entities. CLIA is government controlled and CAP is private. Whereas CAP recently decided to separate grossing into two different complexities, CLIA has not. Bottom line...if you are CLIA licensed you MUST follow the CLIA standard. If not, then you can do whatever you like. Charles Embrey Jr., PA(ASCP)" I'm not sure I'm sold on this yet, b/c also in the CLIA regs it states in sec. 493.559 about exemption from CLIA, via using a private accreditation agency (which is what CAP is), and states clearly that this agency can only be deemed acceptable if their requirements for the laboratory are equivalent or more stringent and the laboratory would meet CLIA requirements if the laboratory had not been granted deemed status. Ya'll are right, this is a mess! So, wouldn't this essentially mean that according to CLIA, since CAP is an acceptable private agency, they are giving the ok for CAP's decision on this? Wouldn't CAP be unallowed to do what they do if it was less stringent than CLIA? I also find nothing in the entire CLIA 88' that indicates that this would not be ok. In fact, CLIA has a criteria for categorization of complexity: http://www.fda.gov/cdrh/clia/categorization.html . And in their database of tests that they have said has to absolutely be a certain way, I find no reference to tissue description, gross examination, tissue examination, or processing. Have I stumped anyone yet, or is there some more things I am missing? I really appreciate all ya'lls willingness to re-visit this issue again. I guess I am a stuborn monkey! Thanks, Vince Van Cleave, BS, PA(ASCP), HTL, HT Laboratory Manager and Pathologist Assistant (325)670-6528 Clinical Pathology Associates Abilene, TX PRIVILEGED AND CONFIDENTIAL NOTICE: The information contained in this e-mail may be confidential and/or privileged. This e-mail is intended to be reviewed initially by only the individual named above. If the reader of this e-mail is not the intended recipient or a representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail, or the information contained herein, is prohibited. If you have received this e-mail in error, notify the above sender, send the message back and then delete it. Thank you for your assistance. Hendrick Health System 1900 Pine St. Abilene, Texas 79601 - 325.670.2000 79601 - 325.670.2000 From CEgan <@t> healthsearchgroup.com Wed Nov 7 16:28:23 2007 From: CEgan <@t> healthsearchgroup.com (Carolyn Egan) Date: Wed Nov 7 16:28:35 2007 Subject: [Histonet] Mailing lists Message-ID: Please remove my name from the mailing list. Thank you for your courtesy. Sincerely, Carolyn Egan Executive Consultant The Health Search Group (914) 941 - 6107, x118 e-mail: cegan@healthsearchgroup.com From pierre.chaumat <@t> alphelys.com Wed Nov 7 17:41:52 2007 From: pierre.chaumat <@t> alphelys.com (Pierre CHAUMAT) Date: Wed Nov 7 17:42:03 2007 Subject: [Histonet] TMA arrayers. Does anyone have a favorite model ? In-Reply-To: <200711061504.lA6F496r004558@smtp09.msg.oleane.net> Message-ID: <0MKwtQ-1IpuWs1nVw-0001gc@mrelayeu.kundenserver.de> Dear all, Greg, you mentionned Boekeler but on their web site, I could not find any tissuer arrayer. Would you have a link to an exact page ? Thanks Pierre -----Message d'origine----- De : histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] De la part de histonet-request@lists.utsouthwestern.edu Envoy? : mardi 6 novembre 2007 16:04 ? : histonet@lists.utsouthwestern.edu Objet : Histonet Digest, Vol 48, Issue 6 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. TMA arrayers. Does anyone have a favorite model? (Hugh Luk) 2. Deplasticizing MMA sections (Nancy W. Troiano) 3. Histo, cyto job in NY (Xorren@aol.com) 4. Re: TMA arrayers. Does anyone have a favorite model? (Greg Dobbin) 5. NSH Report (Breeden, Sara) 6. Re: TMA arrayers. Does anyone have a favorite model? (Thomas Pier) 7. Re: Histonet Digest, Vol 48, Issue 5 (Boneslides@aol.com) 8. RE: deplasticizing PMMA sections for staining (Monfils, Paul) 9. Re: TMA arrayers. Web link (Greg Dobbin) 10. NSH show (Rathborne, Toni) 11. RE: Mucin overstained with Hematoxylin in G.I. specimens (Charles.Embrey) 12. RE: Mucin overstained with Hematoxylin in G.I. specimens (Bonnie Whitaker) 13. RE: Mucin overstained with Hematoxylin in G.I. specimens (Douglas D Deltour) 14. RE: deplasticizing PMMA sections for staining (Yaskovich, Ruth A (NIH/NIDCR) [E]) 15. NSH (Bernice Frederick) 16. RE: Mucin overstained with Hematoxylin in G.I. specimens (LuAnn Anderson) 17. how to keep thick sections on the slides after deparaffin (FU,DONTAO) 18. Re: Mucin overstained with Hematoxylin in G.I. specimens (DNon) 19. RE: how to keep thick sections on the slides after deparaffin (Kemlo Rogerson) 20. FYI - Brainbow (Barbara Webb) 21. RE: Histo, cyto job in NY (Lee & Peggy Wenk) 22. RELIA's Histology Opportunities Update 11-06-07 (Pam Barker) 23. RE: Histo, cyto job in NY (James Watson) ---------------------------------------------------------------------- Message: 1 Date: Mon, 5 Nov 2007 08:22:56 -1000 From: Hugh Luk Subject: [Histonet] TMA arrayers. Does anyone have a favorite model? To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct ------------------------------ Message: 2 Date: Mon, 05 Nov 2007 13:25:52 -0500 From: "Nancy W. Troiano" Subject: [Histonet] Deplasticizing MMA sections To: histonet@lists.utsouthwestern.edu Message-ID: <5.2.1.1.2.20071105132352.00c3f668@email.med.yale.edu> Content-Type: text/plain; format=flowed; charset=us-ascii Try soaking the slides in two 5 minute changes of acetone (100%) to deplastify or you can deplastify in 2 changes of Cellosolve (Fisher, cat. #E181-4) 25 minutes each, followed by 5 minutes in 70% ethanol, 40% ethanol, then distilled water for five minutes prior to doing your staining. ------------------------------ Message: 3 Date: Mon, 5 Nov 2007 13:44:13 EST From: Xorren@aol.com Subject: [Histonet] Histo, cyto job in NY To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Hello all ! Does anyone know of any histology or cytology prep job in the NY area. I have have 15 years in the field and hope someone has some info. you can respond @ xorren@aol.com or call (516) 671-0907 have a great day !!!!! ************************************** See what's new at http://www.aol.com ------------------------------ Message: 4 Date: Mon, 05 Nov 2007 14:44:33 -0400 From: "Greg Dobbin" Subject: Re: [Histonet] TMA arrayers. Does anyone have a favorite model? To: Message-ID: Content-Type: text/plain; charset=US-ASCII I am investigating an instrument that I saw at NSH by RMC Products. Model no. is KIN-1. The picture on the web site does not match exactly the unit they had at NSH but they could send you the same broshure they were passing out at their booth (it must be available as a pdf). Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> hlukey@msn.com 11/5/2007 2:22 PM >>> I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct__ _____________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. ------------------------------ Message: 5 Date: Mon, 5 Nov 2007 11:50:55 -0700 From: "Breeden, Sara" Subject: [Histonet] NSH Report To: Message-ID: <4D14F0FC9316DD41972D5F03C070908B8F4725@nmdamailsvr.nmda.ad.nmsu.edu> Content-Type: text/plain; charset="us-ascii" Okay, I'm back from my week at NSH and this is my report. 1. The NSH staff is awesome. Aubrey, Carrie and the entire NSH staff pulled off yet another flawless meeting. We can't thank NSH enough for all they do! 2. Joe Nocito is alive and well. I met him. He lives and he wasn't wearing a flak jacket. 3. Our vendors are magnificent - supportive, creative, helpful and approachable. Kudos!! 4. Each one of us needs to recruit NSH members - there is strength in numbers. NSH is our connection in SO many ways! 5. Ada Feldman makes a terrific clown! 6. Did you know that histologists are the ONLY technicians not recognized by the US Government as healthcare professionals? 7. After 3 days of walking the 6 blocks from hotel to convention center, I discovered the free bus. 8. There must be a "message board" at next year's meeting so we can find each other if we need to. 9. I want to know what vitamins Peggy Wenk takes. Seriously. Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 ------------------------------ Message: 6 Date: Mon, 05 Nov 2007 12:55:36 -0600 From: "Thomas Pier" Subject: Re: [Histonet] TMA arrayers. Does anyone have a favorite model? To: , Message-ID: <472F12C8020000DF00002B3D@gwmail.medicine.wisc.edu> Content-Type: text/plain; charset=US-ASCII I haven't used any other instruments, but I've been happy with the Beecher MTA-1. Tom Pier >>> Hugh Luk 11/05/07 12:22 PM >>> I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct__ _____________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Mon, 5 Nov 2007 14:10:40 EST From: Boneslides@aol.com Subject: [Histonet] Re: Histonet Digest, Vol 48, Issue 5 To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" I de-plasticize my PMMA sections in methyl acetate...takes about 30 minutes, then rehydrate thru several changes of absolute, 95% and 70% alcohol, rinse in water, stain as desired. Diane M. Mahovlic, HT, MLT(ASCP) Orthopedic Pathology & Biomaterials Laboratory Department of Anatomic Pathology The Cleveland Clinic Foundation 9500 Euclid Avenue- L30 Cleveland, Ohio 44195 216-444-0166 ************************************** See what's new at http://www.aol.com ------------------------------ Message: 8 Date: Mon, 5 Nov 2007 14:22:08 -0500 From: "Monfils, Paul" Subject: RE: [Histonet] deplasticizing PMMA sections for staining To: Message-ID: <4EBFF65383B74D49995298C4976D1D5E273CF6@LSRIEXCH1.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" In my experience, xylene is a rather poor solvent for PMMA. It works much faster at 60 degrees, but don't try heating flammable solvents in your oven unless you are certain it is certified explosion-proof (most paraffin ovens are not). Even then, unless you have the oven in or at least next to a fume hood, you'll get a lot of xylene vapors in the air as a result of heating it. I deplasticise PMMA with 2-methoxyethyl acetate, which only takes a couple of hours at room temperature. This has a rather strong though not unpleasant odor, and should be used in the hood. ------------------------------ Message: 9 Date: Mon, 05 Nov 2007 16:12:32 -0400 From: "Greg Dobbin" Subject: Re: [Histonet] TMA arrayers. Web link To: Message-ID: Content-Type: text/plain; charset=US-ASCII Sorry. I see the company name is actually Boeckeler and the website is www.boeckeler.com Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> "Greg Dobbin" 11/5/2007 2:44 PM >>> I am investigating an instrument that I saw at NSH by RMC Products. Model no. is KIN-1. The picture on the web site does not match exactly the unit they had at NSH but they could send you the same broshure they were passing out at their booth (it must be available as a pdf). Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> hlukey@msn.com 11/5/2007 2:22 PM >>> I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct__ _____________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. ------------------------------ Message: 10 Date: Mon, 5 Nov 2007 15:30:49 -0500 From: "Rathborne, Toni" Subject: [Histonet] NSH show To: Message-ID: Content-Type: text/plain; charset="utf-8" How was the show? I heard there were a lot of vendors there. What about the lectures/workshops? I was hoping to go, so many relevant topics offered. A friend of mine was there and mentioned that Ventana had some sort of secretive booth. What was that all about? Is it a new IHC stainer? Or part of their whole "lab platform"? Toni CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. Somerset Medical Center is proud to receive the Somerset County Business Partnership's 2006 Quality of Life Award. ------------------------------ Message: 11 Date: Mon, 5 Nov 2007 15:08:54 -0600 From: "Charles.Embrey" Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "DNon" , Message-ID: <44780C571F28624DBB446DE55C4D733A1FE565@EXCHANGEBE1.carle.com> Content-Type: text/plain; charset="us-ascii" I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Mon, 5 Nov 2007 15:22:45 -0600 From: "Bonnie Whitaker" Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "'Charles.Embrey'" , "'DNon'" , Message-ID: <003501c81ff2$02602060$3601a8c0@brownpathology.net> Content-Type: text/plain; charset="us-ascii" Hi All, I had the same type incident (with the increase in mucin staining with the Surgipath hematoxylin), but it was a one time thing that we attributed to a larger than normal volume of H&E's in that particular time period. We increased the frequency that solutions are changed slightly, and haven't had another problem. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, November 05, 2007 3:09 PM To: DNon; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@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: 13 Date: Mon, 5 Nov 2007 16:26:45 -0500 From: "Douglas D Deltour" Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "'Charles.Embrey'" , "'DNon'" , Message-ID: Content-Type: text/plain; charset="US-ASCII" Charles, We also are having the same issue with the Surgipath Gills III. This issue also started last month. Very interesting. Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, November 05, 2007 4:09 PM To: DNon; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@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: 14 Date: Mon, 5 Nov 2007 16:43:34 -0500 From: "Yaskovich, Ruth A (NIH/NIDCR) [E]" Subject: RE: [Histonet] deplasticizing PMMA sections for staining To: "John Baker" , Message-ID: Content-Type: text/plain; charset="us-ascii" John, You can soak them in 3 changes of acetone for 15 minutes each. Put them in the first acetone right out of the oven. Ruth Yaskovich National Institutes of Health National Institute of Dental and Craniofacial Research -----Original Message----- From: John Baker [mailto:bakerj@umich.edu] Sent: Monday, November 05, 2007 12:07 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] deplasticizing PMMA sections for staining > > > Hello All, We are sectioning PMMA embedded rat femur at 5 microns on > the Polycut. Then we are deplasticizing for H & E and Trichrome > stains. It has been taking over 2 weeks with several changes of xylene > to get them done and still some remnants of plastic there. I see in > Sheehan's Practice and Theory of Histotechnology that in several > protocols they deplasticize in xylene for 4 hours at 55 degrees C. > The flash point of xylene is 25 degree C. Is it safe in covered > staining dishes to actually do this? Also, we use Mayer's Hematoxylin > and 2% Eosin to stain. What H & E protocol do you use for bone? Any > suggestions on how to speed up the deplasticizing process welcome. > Does anyone have Diane Sterchi's and Gayle Callis's email addresses, > lost them? > Thanks, John > > > > > John A. Baker The University of Michigan Orthopaedic Research Laboratories Histology Unit 109 Zina Pitcher Place, 2218 BSRB Ann Arbor, MI 48109-2200 734-936-1635 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Mon, 5 Nov 2007 15:45:56 -0600 From: "Bernice Frederick" Subject: [Histonet] NSH To: Message-ID: <000001c81ff5$42a1aec0$d00f7ca5@lurie.northwestern.edu> Content-Type: text/plain; charset="us-ascii" This was my first meeting and whereas I enjoyed myself tremendously I only had 2 gripes 1. There should be a block of time set aside to see the vendors. Most of us were in workshops for a lot of the time and had to do quick run up to the vendors. 2. I was surprised water was only provided for the speakers. It was so dry I was going through 36-48 oz of water every day and at 2$ a pop... If I'd known, I'd have run off to Walgreens or somewhere. I do plan to be back and really enjoyed everybody and everything. Missed meeting Joe though I thought we had arranged for that (tee hee). Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 ------------------------------ Message: 16 Date: Mon, 05 Nov 2007 15:51:47 -0600 From: LuAnn Anderson Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "Charles.Embrey" , "DNon" , Message-ID: Content-Type: text/plain; charset="us-ascii"; format=flowed I too have had problems recently with Surgipath hematoxylin in brain tissue (not mucin). They replaced what I had with a new bottle, but I am seeing the same problems. I believe the formula has been changed to eliminate mercury--don't know if this is the basis of the problems or not--but I believe that is when I started having problems. They have a "new" hematoxylin (560) and eosin (515), but our pathologists did not care for those either. I have been using Surgipath for 14 years and just started having staining problems with it. LuAnn Anderson Neuropthology Lab University of Minnesota At 03:08 PM 11/5/2007, Charles.Embrey wrote: >I find this post most interesting. After using Surgipath Gills III for >the past 7 years we started having a problem with blue mucin just last >month. I corrected the problem by adding GAA but I now have to monitor >the pH on a daily basis. It is strange that after all these years I >would suddenly have this problem and then read about another lab having >the same difficulty with Surgipath Hematoxylin starting about the same >timeframe. Has Surgipath changed something? > >Charles Embrey Jr., PA(ASCP) >Histology Manager, Carle Clinic > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon >Sent: Saturday, November 03, 2007 9:42 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens > >Hello Histonet, > > Over the past several weeks our lab has had problems with >hematoxylin overstaining in the mucin of our gastric specimens. The >degree of hematoxylin overstaining is alarming in the mucin of the >gastric specimens but absent or negligible in other specimens. One >pathologist reviewing the slides indicated some of the overstained >specimens appeared to be inadequately fixed. We have addressed that >issue which showed some improvement, but the problem persists. We are >making changes to increase fixation time (although the specimens now >appear adequately fixed yet retain a large measure of the overstaining), >but any further suggestions for corrective action would be much >appreciated. > > We've been hesitant to increase acid alcohol clearing time because >our other specimens are staining well with the current protocol. > >We are using the following reagents and stain times: > >- Surgipath specimen containers pre-filled with 10% neutral buffered >Formalin. >- Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running >water washes >- .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water >wash >- Surgipath Scott's Tap Water for 1 min followed by running water wash > >Sakura automated stainer set to agitate ( up and down dipping of racks ) >once every 2 seconds, which is the fastest speed available on it. > > >Dick Non >Pathology Department >Ocean County Medical Lab >Brick, New Jersey >_______________________________________________ >Histonet mailing list >Histonet@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: 17 Date: Mon, 5 Nov 2007 16:52:46 -0500 From: "FU,DONTAO" Subject: [Histonet] how to keep thick sections on the slides after deparaffin To: Message-ID: <81FEA2B93DC53C4B8CF3667707CA592A0DCE8C@path-exchange.ad.ufl.edu> Content-Type: text/plain; charset="us-ascii" Hi, I met a big problem in keeping thick sections on the slides after deparaffin recently. I cut a 70um-thick mouse retina section on superfrost plus Gold slides, leaving it dry for 2days. Before deparaffin, I put the slide in the 65C oven for 30 min to let it melt, then I did the general deparaffin procedure. Unfortunately, the section fell off from the slide. Does anyone have any experience on deparaffin thick sections? Any suggestions? Thank you, Ann Dongtao Fu MD Ph.D Lab Manager Molecular Pathology core Dept. of Pathology University of Florida Lab Phone: 352-273-7752 FAX: 352-273-7753 Rm: D11-50 ------------------------------ Message: 18 Date: Mon, 05 Nov 2007 18:51:18 -0500 From: "DNon" Subject: Re: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "Charles.Embrey" Cc: histonet@lists.utsouthwestern.edu Message-ID: <000601c82006$c3081ac0$1f0aa8c0@cyto.ocml.com> Content-Type: text/plain; format=flowed; charset=iso-8859-1; reply-type=original I spoke with Surgipath's QC department today and asked if there was a change in their Gill's III formulation. According to Mike Urban at Surgipath, nothing has changed. Nonetheless, he is going to run additional tests on the lot number in question and will get back to me. I find your post very interesting indeed as our lab has used Surgipath Gill's III for the last 10 years. They ( Surgipath) will also be sending me a gallon of Gill's III from a different lot. If it doesn't show improvement, we'll either lower the ph similar to what you did, or switch to Harris hematoxylin. ----- Original Message ----- From: "Charles.Embrey" To: "DNon" ; Sent: Monday, November 05, 2007 4:08 PM Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Tue, 6 Nov 2007 08:24:24 -0000 From: "Kemlo Rogerson" Subject: RE: [Histonet] how to keep thick sections on the slides after deparaffin To: "FU,DONTAO" , Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F0E6@wahtntex2.waht.swest.nhs.uk> Content-Type: text/plain; charset="us-ascii" Hi, I met a big problem in keeping thick sections on the slides after deparaffin recently. I cut a 70um-thick mouse retina section on superfrost plus Gold slides, leaving it dry for 2days. Before deparaffin, I put the slide in the 65C oven for 30 min to let it melt, then I did the general deparaffin procedure. Unfortunately, the section fell off from the slide. Does anyone have any experience on deparaffin thick sections? Any suggestions? Thank you, You could use bovine albumin to coat the slides (I think 1% but I'm not sure) then dry at 37 degrees for those two days then into the oven at 60 degrees (ish). Think that sort of sets the section onto the slide but you need to get the concentration correct as the protein could stain with some techniques if its too concentrated. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation ------------------------------ Message: 20 Date: Tue, 06 Nov 2007 10:07:04 GMT From: "Barbara Webb" Subject: [Histonet] FYI - Brainbow To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="utf-8" Looking at the fuure? http://www.nytimes.com/2007/11/06/health/research/06brai.html?_r=1&ref=scien ce&oref=slogin ------------------------------ Message: 21 Date: Tue, 6 Nov 2007 05:20:07 -0500 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] Histo, cyto job in NY To: , Message-ID: <000801c8205e$9b559130$0202a8c0@HPPav2> Content-Type: text/plain; charset="us-ascii" If you are looking for a job as a histotech in New York, it's not going to happen. The NY legislators have made a licensure law that only certified med techs can be histotechs. Those already working as histotechs in NY can continue to work in NY. However, any certified histotech moving into NY cannot work as a histotech until they take all the med tech courses and pass the med tech certification exam. NY will not recognize the NAACLS histotech program graduates from SUNY, the only accredited HT program in NY. (SUNY will be closing, as it's impossible to offer all med tech courses and all histotech courses, all in a 2 year program.) NY will not recognize those earning their ASCP HT or HTL certification through the associate degree on-the-job training route. ASCP, NY histotech society, and the NY pathologist groups are lobbying for changes, but the legislators are not listening. NSH has weighed in with ASCP, and the NY pathologists are coming to NSH to see what NSH can do, too. (That's where Vinnie DelaSperanza's (NSH presidents) comments on needing more members come into play. If ASCP and pathologist groups aren't swaying the legislators, then NSH with our fewer numbers are not going to help. Histotechs need to join NSH and ASCP, to histotechs have more numbers and will be listened to better.) The original draft of the bill, according to Vinnie, included histotechs with the appropriate information on training and certification and job responsibilities. The bill got changed over the years, and went unnoticed by NY histotechs. The bill took years (decade+) to pass, so the legislators don't want to change anything on the law at this stage. It may take years to correct. At this time, the med tech programs do not have classes in the theory or practical aspects of histotechnology. I don't know if they are planning on adding them to the med tech curriculum. In the mean time, NY histotechs will be retiring and leaving the field. Workloads will increase as the patient populations ages and technology requires more tissue tests. And there will be fewer and fewer NY histotechs to do these jobs. It's a histology crisis in the making. Peggy A. 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 Xorren@aol.com Sent: Monday, November 05, 2007 1:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histo, cyto job in NY Hello all ! Does anyone know of any histology or cytology prep job in the NY area. I have have 15 years in the field and hope someone has some info. you can respond @ xorren@aol.com or call (516) 671-0907 have a great day !!!!! ************************************** See what's new at http://www.aol.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 22 Date: Tue, 6 Nov 2007 08:21:20 -0500 From: "Pam Barker" Subject: [Histonet] RELIA's Histology Opportunities Update 11-06-07 To: "'Histonet'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi Histonetters! I have been reading some of the posts on the NSH in Denver. What about everybody else, did you make it to the NSH in Denver? If so how was it? What was your favorite thing about it? I havent been since the convention in 2005 in Ft. Lauderdale. Did you have a get together for the histonet? We did at the Ft. Lauderdale convention it was a small turnout but we had fun. I really enjoyed meeting people, learning new things and the party at the Hard Rock Casino. I am planning on attending the 2008 meeting in Pittsburgh. Will I see you there? Here is a list of my current openings all of the positions are full time permanent positions. They are M-F dayshift opportunities and my clients offer excellent compensation, benefits and relocation assistance. And they are ready to interview and hire right away. ****RELIA Spotlight Opportunities**** I am working with a client in the Philadelphia, PA area that has 2 unique and exciting opportunities. These are nonclinical positions. BioSciences Product Manager Histologist Technical and Field Support ********************************* Histology Managers/Supervisors needed in: Dallas, TX Austin, TX Philadelphia, PA Valencia, CA Cape Cod, MA Histotechnicians/Histotechnologists needed in: Phoenix, AZ Atlanta, GA San Francisco, CA Los Angeles, CA Valencia, CA Orlando, FL St. Petersburg, FL Fredericksburg, VA Dallas, TX Waco, TX Austin, TX Philadelphia, PA Pittsburgh, PA If you know of anyone else who might be interested in any of these positions or might like to subscribe to RELIAs Histology Careers Bulletin please feel free to pass this e-mail along to them. Remember it never hurts to look!!! Thanks, Pam 877-60 RELIA (877-607-3542) Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net www.myspace.com/pamatrelia ------------------------------ Message: 23 Date: Tue, 6 Nov 2007 06:41:31 -0800 From: "James Watson" Subject: RE: [Histonet] Histo, cyto job in NY To: , , Message-ID: Content-Type: text/plain; charset="iso-8859-1" At NSH we were told by someone that was in contact with the legistators that wrote the bill that this did not pass and that they were rewriting the bill. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Lee & Peggy Wenk Sent: Tue 11/6/2007 2:20 AM To: Xorren@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histo, cyto job in NY If you are looking for a job as a histotech in New York, it's not going to happen. The NY legislators have made a licensure law that only certified med techs can be histotechs. Those already working as histotechs in NY can continue to work in NY. However, any certified histotech moving into NY cannot work as a histotech until they take all the med tech courses and pass the med tech certification exam. NY will not recognize the NAACLS histotech program graduates from SUNY, the only accredited HT program in NY. (SUNY will be closing, as it's impossible to offer all med tech courses and all histotech courses, all in a 2 year program.) NY will not recognize those earning their ASCP HT or HTL certification through the associate degree on-the-job training route. ASCP, NY histotech society, and the NY pathologist groups are lobbying for changes, but the legislators are not listening. NSH has weighed in with ASCP, and the NY pathologists are coming to NSH to see what NSH can do, too. (That's where Vinnie DelaSperanza's (NSH presidents) comments on needing more members come into play. If ASCP and pathologist groups aren't swaying the legislators, then NSH with our fewer numbers are not going to help. Histotechs need to join NSH and ASCP, to histotechs have more numbers and will be listened to better.) The original draft of the bill, according to Vinnie, included histotechs with the appropriate information on training and certification and job responsibilities. The bill got changed over the years, and went unnoticed by NY histotechs. The bill took years (decade+) to pass, so the legislators don't want to change anything on the law at this stage. It may take years to correct. At this time, the med tech programs do not have classes in the theory or practical aspects of histotechnology. I don't know if they are planning on adding them to the med tech curriculum. In the mean time, NY histotechs will be retiring and leaving the field. Workloads will increase as the patient populations ages and technology requires more tissue tests. And there will be fewer and fewer NY histotechs to do these jobs. It's a histology crisis in the making. Peggy A. 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 Xorren@aol.com Sent: Monday, November 05, 2007 1:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histo, cyto job in NY Hello all ! Does anyone know of any histology or cytology prep job in the NY area. I have have 15 years in the field and hope someone has some info. you can respond @ xorren@aol.com or call (516) 671-0907 have a great day !!!!! ************************************** See what's new at http://www.aol.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 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 6 *************************************** From mtarango <@t> nvcancer.org Wed Nov 7 18:19:58 2007 From: mtarango <@t> nvcancer.org (Tarango, Mark) Date: Wed Nov 7 18:20:19 2007 Subject: [Histonet] viability stain for 450 micron brain slices In-Reply-To: <464094433.180371194446612705.JavaMail.osg@osgjas03.cns.ufl.edu> Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F6B@NVCIEXCH02.NVCI.org> The first thing that comes to mind is Trypan blue, but that would be for regular light microscopy. If you want to do florescent you might try 7AAD or propidium idodide. An ethidium bromide / acridine orange combo might be good too. I'm not sure about fixing the staining. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of HENKEN,KIRSTEN RUTH Sent: Wednesday, November 07, 2007 6:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] viability stain for 450 micron brain slices I am looking for a staining technique to test the viability of slices of the rat brain after some force-displacement tests on the tissue. As I am an engineer and no chemist I do not know what kind of stain would be appropriate. We keep the slices from the cerebral cortex and the corpus callosumalive alive in a tissue chamber with ACSF that is bubbled with 95% O2/ 5% CO2. The slices have a thickness of 450 micron and there is a fluorescent confocal microscope available. As this microscope is away from the testing site and we do not have the equipment that can keep the tissue alive during imaging, it would be very convenient to fix the tissue after staining. Does anyone of you have experience with a staining technique that is suitable for my experiment? Best, Kirsten Henken _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== From liz <@t> premierlab.com Wed Nov 7 18:38:00 2007 From: liz <@t> premierlab.com (Liz Chlipala) Date: Wed Nov 7 18:38:08 2007 Subject: [Histonet] viability stain for 450 micron brain slices In-Reply-To: <001910BE531548AA9CF0A4DF03FEF8D4@PremierLab.local> References: <464094433.180371194446612705.JavaMail.osg@osgjas03.cns.ufl.edu> <001910BE531548AA9CF0A4DF03FEF8D4@PremierLab.local> Message-ID: Try a live cell/dead cell kit. You see this application used alot on thick sections of cartilage, etc. I think I might have a reference for thick sections of articular cartilage. If you can get away without using fluoresence I would try TTC staining. Just search for TTC and rat brain or something like that, its used quite a bit in the literature. You might also try NADH. I have a protocol for the NADH stain but its for frozen sections and it may work for thick sections. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, CO 80308 phone (303) 735-5001 fax (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory, LLC University of Colorado at Boulder MCDB, Room A3B40 Boulder, CO 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tarango, Mark Sent: Wednesday, November 07, 2007 5:31 PM To: HENKEN,KIRSTEN RUTH; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] viability stain for 450 micron brain slices The first thing that comes to mind is Trypan blue, but that would be for regular light microscopy. If you want to do florescent you might try 7AAD or propidium idodide. An ethidium bromide / acridine orange combo might be good too. I'm not sure about fixing the staining. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of HENKEN,KIRSTEN RUTH Sent: Wednesday, November 07, 2007 6:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] viability stain for 450 micron brain slices I am looking for a staining technique to test the viability of slices of the rat brain after some force-displacement tests on the tissue. As I am an engineer and no chemist I do not know what kind of stain would be appropriate. We keep the slices from the cerebral cortex and the corpus callosumalive alive in a tissue chamber with ACSF that is bubbled with 95% O2/ 5% CO2. The slices have a thickness of 450 micron and there is a fluorescent confocal microscope available. As this microscope is away from the testing site and we do not have the equipment that can keep the tissue alive during imaging, it would be very convenient to fix the tissue after staining. Does anyone of you have experience with a staining technique that is suitable for my experiment? Best, Kirsten Henken _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------ ------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ======================================================================== ====== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CIngles <@t> uwhealth.org Wed Nov 7 18:35:22 2007 From: CIngles <@t> uwhealth.org (Ingles Claire) Date: Wed Nov 7 18:39:21 2007 Subject: [Histonet] Still not sure about this low complexity or not References: <740F77F6594C20458BDC2A4A500D8D4150163F@EMAIL.hhs.hendrickhealth.org> Message-ID: <08A0A863637F1349BBFD83A96B27A50A12009A@uwhis-xchng3.uwhis.hosp.wisc.edu> Vince: I believe this to be interpreted as by "using a private accreditation agency..." they mean having CAP DO the accreditation survey and not CLIA. If that were the case, then the CLIA rules can just be thrown out anyway. Anybody else willing to wade in? Claire ________________________________ Charles Embrey Jr., PA(ASCP)" I'm not sure I'm sold on this yet, b/c also in the CLIA regs it states in sec. 493.559 about exemption from CLIA, via using a private accreditation agency (which is what CAP is), and states clearly that this agency can only be deemed acceptable if their requirements for the laboratory are equivalent or more stringent and the laboratory would meet CLIA requirements if the laboratory had not been granted deemed status. Ya'll are right, this is a mess! So, wouldn't this essentially mean that according to CLIA, since CAP is an acceptable private agency, they are giving the ok for CAP's decision on this? Wouldn't CAP be unallowed to do what they do if it was less stringent than CLIA? I also find nothing in the entire CLIA 88' that indicates that this would not be ok. In fact, CLIA has a criteria for categorization of complexity: http://www.fda.gov/cdrh/clia/categorization.html . And in their database of tests that they have said has to absolutely be a certain way, I find no reference to tissue description, gross examination, tissue examination, or processing. Have I stumped anyone yet, or is there some more things I am missing? I really appreciate all ya'lls willingness to re-visit this issue again. I guess I am a stuborn monkey! Thanks, Vince Van Cleave, BS, PA(ASCP), HTL, HT Laboratory Manager and Pathologist Assistant (325)670-6528 Clinical Pathology Associates Abilene, TX From dis.tox <@t> suven.com Thu Nov 8 00:41:41 2007 From: dis.tox <@t> suven.com (Toxicology) Date: Thu Nov 8 00:34:39 2007 Subject: [Histonet] Artefacts Message-ID: Dear all, I am facing a problem in Hematoxylin and Eosin staining. There are several faint stained spots on sections and because of that its difficuly to interpret the lesions. I am unable to sort out the problem. Can any one help me? -------------------------Email Disclaimer--------------------------------- This e-mail may contain confidential and/or privileged information. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden. From arnie.jimenez <@t> vel-lab.com Thu Nov 8 00:37:36 2007 From: arnie.jimenez <@t> vel-lab.com (arnie.jimenez@vel-lab.com) Date: Thu Nov 8 00:39:49 2007 Subject: [Histonet] Re: Away from Office Message-ID: <20071108063736.19107.qmail@plesk5.hostgator.com> Thank you for contacting Vel-Lab Research. We will be out of the lab starting Nov. 7th and will return on Nov. 12th. We will check e-mail regularly but will be unable to respond. Please do not mail any packages to our lab until Nov. 12th. Regards, Arnie Jimenez Vel-Lab Research From dis.tox <@t> suven.com Thu Nov 8 00:50:15 2007 From: dis.tox <@t> suven.com (Toxicology) Date: Thu Nov 8 00:46:42 2007 Subject: [Histonet] Fwd: Artefacts Message-ID: Dear all, I am facing a problem in Hematoxylin and Eosin staining. There are several faint stained spots on sections and because of that its difficuly to interpret the lesions. I am unable to sort out the problem. Can any one help me? I have posted a picture to www.histonet.org. named artefact. -----Original Message----- From: "Toxicology" To: histonet@lists.utsouthwestern.edu Date: Thu, 08 Nov 2007 12:11:41 +0530 Subject: Artefacts Dear all, I am facing a problem in Hematoxylin and Eosin staining. There are several faint stained spots on sections and because of that its difficuly to interpret the lesions. I am unable to sort out the problem. Can any one help me? -------------------------Email Disclaimer--------------------------------- This e-mail may contain confidential and/or privileged information. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden. From Kemlo.Rogerson <@t> waht.swest.nhs.uk Thu Nov 8 03:20:21 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu Nov 8 03:20:32 2007 Subject: [Histonet] Artefacts Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F121@wahtntex2.waht.swest.nhs.uk> Ah the spotty H&E problem again. 1) Inadequate dewaxing. 2) Inadequate fixation. 3) Inadequate processing. Not necessarily in that order. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From pierre.chaumat <@t> alphelys.com Thu Nov 8 04:06:53 2007 From: pierre.chaumat <@t> alphelys.com (Pierre CHAUMAT) Date: Thu Nov 8 04:06:59 2007 Subject: [Histonet] RE: Formaline-free fixative In-Reply-To: <200711080634.lA86YoGm027387@smtp13.msg.oleane.net> Message-ID: <0ML21M-1Iq4Hj1GWe-00029l@mrelayeu.kundenserver.de> Hello Histonet, Has anyone tested alternative solution to formaline fixation ? Has anyone switched to a new tissue fixative ? I would like to share some experience. Kind regards Pierre From tkngflght <@t> yahoo.com Thu Nov 8 07:03:31 2007 From: tkngflght <@t> yahoo.com (Cheryl R. Kerry) Date: Thu Nov 8 07:03:49 2007 Subject: [Histonet] Anyone want to live in Texas?? Temp-to-perm in TX, SC, CT and more. In-Reply-To: <08A0A863637F1349BBFD83A96B27A50A12009A@uwhis-xchng3.uwhis.hosp.wisc.edu> Message-ID: <01a001c82207$c3b9ac60$6801a8c0@CHERYLSLAPTOP> Hi All- I have FIVE open positions for temp-to-perm, all in the great Lone Star State of Texas (yes, I live in TX and was surprised at how much I LIKE it!). These are all over the state, in different lab types, different shifts and different responsibilities from routine bench to management. I've worked in several of these labs and have had our temps in some of the others--good places, all. The phone call takes about 10 minutes and I don't come across as a used car salesman--it's your life and my job is to help you find something that allows you to be HAPPY in your work! Give me a ring--it is an easy conversation. Cheryl Cheryl R. Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab, one great tech at a time. 281.852.9457 office 281.883.7704 cell 800.756.3309 fax and alternate phone We have a bunch of other temp-to-perms, too, including SC, CT and a few other places on top of about 40 other permanent positions--if you're curious, just ask! From fudo <@t> ufl.edu Thu Nov 8 07:38:39 2007 From: fudo <@t> ufl.edu (FU,DONGTAO) Date: Thu Nov 8 07:38:48 2007 Subject: [Histonet] problem in dual staining of CD4 and FOXP3 Message-ID: <1287148415.447131194529119265.JavaMail.osg@osgjas02.cns.ufl.edu> Hi, all Thank you first for giving me some good suggestions on thick-section question I posted last time. Now I met another problem when I did CD4 and FOXP3 dual staining using murine fresh frozen spleen. If I did single staining, both antibodies worked very well. However if I did dual staining, I could only get good result from the first antibody. The second one did not work at all(I mean no specific staining). I used CD4 from BD(rat anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. I think there might be something wrong with my serum block(I used normal rat serum block) before I added secondary antibody. Or any other serum block I need to add to decrease the non-specific binding which I have not done yet. Does anyone can give me some suggestions according to my protocol below? How can I get specific staining of the secondary (primary)antibody? Thank you, Below is the simple protocol I used for dual staining: 1. 2% normal goat serum block 20 min 2. 1* antibody CD4 1:750 in diluent O/N 4C 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT 4. Serum block: 5% normal rat serum 30 min 5. 1* antibody FOXP3 1:100 in diluent 1h RT 6. Secondary AF488 Donkey anti-rat in TBS 1h RT Use 1xTBS as wash buffer. Before staining, fix tissue in -20C Aceton for 5 min, then airdry. Ann Dongtao Fu MD Ph.D Lab Manager Molecular Pathology core Dept. of Pathology University of Florida 1600 SW Archer Road Gainesville, FL 32610 Lab Phone: 352-273-7752 FAX: 352-273-7753 Rm: D11-50 From gvdobbin <@t> ihis.org Thu Nov 8 07:39:12 2007 From: gvdobbin <@t> ihis.org (Greg Dobbin) Date: Thu Nov 8 07:39:29 2007 Subject: [Histonet] TISSUE MICROARRAYER-LINK Message-ID: http://www.rmcproducts.com/cms/?ParentItemIDList=17933,24155,24156&ItemID=100258 Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. From hfedor <@t> jhmi.edu Thu Nov 8 07:39:51 2007 From: hfedor <@t> jhmi.edu (Helen Fedor) Date: Thu Nov 8 07:40:11 2007 Subject: [Histonet] TMA arrayers. Does anyone have a favorite model ? In-Reply-To: <0MKwtQ-1IpuWs1nVw-0001gc@mrelayeu.kundenserver.de> References: <200711061504.lA6F496r004558@smtp09.msg.oleane.net> <0MKwtQ-1IpuWs1nVw-0001gc@mrelayeu.kundenserver.de> Message-ID: <4732CB57.61A1.0088.3@jhmi.edu> http://www.rmcproducts.com/cms/index.cfm/path/17933/25999/24156/100258/ Helen L. Fedor B.S. Johns Hopkins University Pathology Department 600 N Wolfe St Marburg Room 406 Baltimore MD 21287 email: hfedor@jhmi.edu Phone: 410 614-1660 Pager: 410 283-3419 WARNING: E-mail sent over the Internet is not secure. Information sent by e-mail may not remain confidential. DISCLAIMER: This e-mail is intended only for the individual to whom it is addressed. It may be used only in accordance with applicable laws. If you received this e-mail by mistake, notify the sender and destroy the e-mail. >>> Pierre CHAUMAT 11/7/2007 6:41 PM >>> Dear all, Greg, you mentionned Boekeler but on their web site, I could not find any tissuer arrayer. Would you have a link to an exact page ? Thanks Pierre -----Message d'origine----- De : histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] De la part de histonet-request@lists.utsouthwestern.edu Envoy? : mardi 6 novembre 2007 16:04 ? : histonet@lists.utsouthwestern.edu Objet : Histonet Digest, Vol 48, Issue 6 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. TMA arrayers. Does anyone have a favorite model? (Hugh Luk) 2. Deplasticizing MMA sections (Nancy W. Troiano) 3. Histo, cyto job in NY (Xorren@aol.com) 4. Re: TMA arrayers. Does anyone have a favorite model? (Greg Dobbin) 5. NSH Report (Breeden, Sara) 6. Re: TMA arrayers. Does anyone have a favorite model? (Thomas Pier) 7. Re: Histonet Digest, Vol 48, Issue 5 (Boneslides@aol.com) 8. RE: deplasticizing PMMA sections for staining (Monfils, Paul) 9. Re: TMA arrayers. Web link (Greg Dobbin) 10. NSH show (Rathborne, Toni) 11. RE: Mucin overstained with Hematoxylin in G.I. specimens (Charles.Embrey) 12. RE: Mucin overstained with Hematoxylin in G.I. specimens (Bonnie Whitaker) 13. RE: Mucin overstained with Hematoxylin in G.I. specimens (Douglas D Deltour) 14. RE: deplasticizing PMMA sections for staining (Yaskovich, Ruth A (NIH/NIDCR) [E]) 15. NSH (Bernice Frederick) 16. RE: Mucin overstained with Hematoxylin in G.I. specimens (LuAnn Anderson) 17. how to keep thick sections on the slides after deparaffin (FU,DONTAO) 18. Re: Mucin overstained with Hematoxylin in G.I. specimens (DNon) 19. RE: how to keep thick sections on the slides after deparaffin (Kemlo Rogerson) 20. FYI - Brainbow (Barbara Webb) 21. RE: Histo, cyto job in NY (Lee & Peggy Wenk) 22. RELIA's Histology Opportunities Update 11-06-07 (Pam Barker) 23. RE: Histo, cyto job in NY (James Watson) ---------------------------------------------------------------------- Message: 1 Date: Mon, 5 Nov 2007 08:22:56 -1000 From: Hugh Luk Subject: [Histonet] TMA arrayers. Does anyone have a favorite model? To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct ------------------------------ Message: 2 Date: Mon, 05 Nov 2007 13:25:52 -0500 From: "Nancy W. Troiano" Subject: [Histonet] Deplasticizing MMA sections To: histonet@lists.utsouthwestern.edu Message-ID: <5.2.1.1.2.20071105132352.00c3f668@email.med.yale.edu> Content-Type: text/plain; format=flowed; charset=us-ascii Try soaking the slides in two 5 minute changes of acetone (100%) to deplastify or you can deplastify in 2 changes of Cellosolve (Fisher, cat. #E181-4) 25 minutes each, followed by 5 minutes in 70% ethanol, 40% ethanol, then distilled water for five minutes prior to doing your staining. ------------------------------ Message: 3 Date: Mon, 5 Nov 2007 13:44:13 EST From: Xorren@aol.com Subject: [Histonet] Histo, cyto job in NY To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Hello all ! Does anyone know of any histology or cytology prep job in the NY area. I have have 15 years in the field and hope someone has some info. you can respond @ xorren@aol.com or call (516) 671-0907 have a great day !!!!! ************************************** See what's new at http://www.aol.com ------------------------------ Message: 4 Date: Mon, 05 Nov 2007 14:44:33 -0400 From: "Greg Dobbin" Subject: Re: [Histonet] TMA arrayers. Does anyone have a favorite model? To: Message-ID: Content-Type: text/plain; charset=US-ASCII I am investigating an instrument that I saw at NSH by RMC Products. Model no. is KIN-1. The picture on the web site does not match exactly the unit they had at NSH but they could send you the same broshure they were passing out at their booth (it must be available as a pdf). Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> hlukey@msn.com 11/5/2007 2:22 PM >>> I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct__ _____________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. ------------------------------ Message: 5 Date: Mon, 5 Nov 2007 11:50:55 -0700 From: "Breeden, Sara" Subject: [Histonet] NSH Report To: Message-ID: <4D14F0FC9316DD41972D5F03C070908B8F4725@nmdamailsvr.nmda.ad.nmsu.edu> Content-Type: text/plain; charset="us-ascii" Okay, I'm back from my week at NSH and this is my report. 1. The NSH staff is awesome. Aubrey, Carrie and the entire NSH staff pulled off yet another flawless meeting. We can't thank NSH enough for all they do! 2. Joe Nocito is alive and well. I met him. He lives and he wasn't wearing a flak jacket. 3. Our vendors are magnificent - supportive, creative, helpful and approachable. Kudos!! 4. Each one of us needs to recruit NSH members - there is strength in numbers. NSH is our connection in SO many ways! 5. Ada Feldman makes a terrific clown! 6. Did you know that histologists are the ONLY technicians not recognized by the US Government as healthcare professionals? 7. After 3 days of walking the 6 blocks from hotel to convention center, I discovered the free bus. 8. There must be a "message board" at next year's meeting so we can find each other if we need to. 9. I want to know what vitamins Peggy Wenk takes. Seriously. Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 ------------------------------ Message: 6 Date: Mon, 05 Nov 2007 12:55:36 -0600 From: "Thomas Pier" Subject: Re: [Histonet] TMA arrayers. Does anyone have a favorite model? To: , Message-ID: <472F12C8020000DF00002B3D@gwmail.medicine.wisc.edu> Content-Type: text/plain; charset=US-ASCII I haven't used any other instruments, but I've been happy with the Beecher MTA-1. Tom Pier >>> Hugh Luk 11/05/07 12:22 PM >>> I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct__ _____________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Mon, 5 Nov 2007 14:10:40 EST From: Boneslides@aol.com Subject: [Histonet] Re: Histonet Digest, Vol 48, Issue 5 To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" I de-plasticize my PMMA sections in methyl acetate...takes about 30 minutes, then rehydrate thru several changes of absolute, 95% and 70% alcohol, rinse in water, stain as desired. Diane M. Mahovlic, HT, MLT(ASCP) Orthopedic Pathology & Biomaterials Laboratory Department of Anatomic Pathology The Cleveland Clinic Foundation 9500 Euclid Avenue- L30 Cleveland, Ohio 44195 216-444-0166 ************************************** See what's new at http://www.aol.com ------------------------------ Message: 8 Date: Mon, 5 Nov 2007 14:22:08 -0500 From: "Monfils, Paul" Subject: RE: [Histonet] deplasticizing PMMA sections for staining To: Message-ID: <4EBFF65383B74D49995298C4976D1D5E273CF6@LSRIEXCH1.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" In my experience, xylene is a rather poor solvent for PMMA. It works much faster at 60 degrees, but don't try heating flammable solvents in your oven unless you are certain it is certified explosion-proof (most paraffin ovens are not). Even then, unless you have the oven in or at least next to a fume hood, you'll get a lot of xylene vapors in the air as a result of heating it. I deplasticise PMMA with 2-methoxyethyl acetate, which only takes a couple of hours at room temperature. This has a rather strong though not unpleasant odor, and should be used in the hood. ------------------------------ Message: 9 Date: Mon, 05 Nov 2007 16:12:32 -0400 From: "Greg Dobbin" Subject: Re: [Histonet] TMA arrayers. Web link To: Message-ID: Content-Type: text/plain; charset=US-ASCII Sorry. I see the company name is actually Boeckeler and the website is www.boeckeler.com Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> "Greg Dobbin" 11/5/2007 2:44 PM >>> I am investigating an instrument that I saw at NSH by RMC Products. Model no. is KIN-1. The picture on the web site does not match exactly the unit they had at NSH but they could send you the same broshure they were passing out at their booth (it must be available as a pdf). Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> hlukey@msn.com 11/5/2007 2:22 PM >>> I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct__ _____________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. ------------------------------ Message: 10 Date: Mon, 5 Nov 2007 15:30:49 -0500 From: "Rathborne, Toni" Subject: [Histonet] NSH show To: Message-ID: Content-Type: text/plain; charset="utf-8" How was the show? I heard there were a lot of vendors there. What about the lectures/workshops? I was hoping to go, so many relevant topics offered. A friend of mine was there and mentioned that Ventana had some sort of secretive booth. What was that all about? Is it a new IHC stainer? Or part of their whole "lab platform"? Toni CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. Somerset Medical Center is proud to receive the Somerset County Business Partnership's 2006 Quality of Life Award. ------------------------------ Message: 11 Date: Mon, 5 Nov 2007 15:08:54 -0600 From: "Charles.Embrey" Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "DNon" , Message-ID: <44780C571F28624DBB446DE55C4D733A1FE565@EXCHANGEBE1.carle.com> Content-Type: text/plain; charset="us-ascii" I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Mon, 5 Nov 2007 15:22:45 -0600 From: "Bonnie Whitaker" Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "'Charles.Embrey'" , "'DNon'" , Message-ID: <003501c81ff2$02602060$3601a8c0@brownpathology.net> Content-Type: text/plain; charset="us-ascii" Hi All, I had the same type incident (with the increase in mucin staining with the Surgipath hematoxylin), but it was a one time thing that we attributed to a larger than normal volume of H&E's in that particular time period. We increased the frequency that solutions are changed slightly, and haven't had another problem. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, November 05, 2007 3:09 PM To: DNon; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@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: 13 Date: Mon, 5 Nov 2007 16:26:45 -0500 From: "Douglas D Deltour" Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "'Charles.Embrey'" , "'DNon'" , Message-ID: Content-Type: text/plain; charset="US-ASCII" Charles, We also are having the same issue with the Surgipath Gills III. This issue also started last month. Very interesting. Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, November 05, 2007 4:09 PM To: DNon; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@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: 14 Date: Mon, 5 Nov 2007 16:43:34 -0500 From: "Yaskovich, Ruth A (NIH/NIDCR) [E]" Subject: RE: [Histonet] deplasticizing PMMA sections for staining To: "John Baker" , Message-ID: Content-Type: text/plain; charset="us-ascii" John, You can soak them in 3 changes of acetone for 15 minutes each. Put them in the first acetone right out of the oven. Ruth Yaskovich National Institutes of Health National Institute of Dental and Craniofacial Research -----Original Message----- From: John Baker [mailto:bakerj@umich.edu] Sent: Monday, November 05, 2007 12:07 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] deplasticizing PMMA sections for staining > > > Hello All, We are sectioning PMMA embedded rat femur at 5 microns on > the Polycut. Then we are deplasticizing for H & E and Trichrome > stains. It has been taking over 2 weeks with several changes of xylene > to get them done and still some remnants of plastic there. I see in > Sheehan's Practice and Theory of Histotechnology that in several > protocols they deplasticize in xylene for 4 hours at 55 degrees C. > The flash point of xylene is 25 degree C. Is it safe in covered > staining dishes to actually do this? Also, we use Mayer's Hematoxylin > and 2% Eosin to stain. What H & E protocol do you use for bone? Any > suggestions on how to speed up the deplasticizing process welcome. > Does anyone have Diane Sterchi's and Gayle Callis's email addresses, > lost them? > Thanks, John > > > > > John A. Baker The University of Michigan Orthopaedic Research Laboratories Histology Unit 109 Zina Pitcher Place, 2218 BSRB Ann Arbor, MI 48109-2200 734-936-1635 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Mon, 5 Nov 2007 15:45:56 -0600 From: "Bernice Frederick" Subject: [Histonet] NSH To: Message-ID: <000001c81ff5$42a1aec0$d00f7ca5@lurie.northwestern.edu> Content-Type: text/plain; charset="us-ascii" This was my first meeting and whereas I enjoyed myself tremendously I only had 2 gripes 1. There should be a block of time set aside to see the vendors. Most of us were in workshops for a lot of the time and had to do quick run up to the vendors. 2. I was surprised water was only provided for the speakers. It was so dry I was going through 36-48 oz of water every day and at 2$ a pop... If I'd known, I'd have run off to Walgreens or somewhere. I do plan to be back and really enjoyed everybody and everything. Missed meeting Joe though I thought we had arranged for that (tee hee). Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 ------------------------------ Message: 16 Date: Mon, 05 Nov 2007 15:51:47 -0600 From: LuAnn Anderson Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "Charles.Embrey" , "DNon" , Message-ID: Content-Type: text/plain; charset="us-ascii"; format=flowed I too have had problems recently with Surgipath hematoxylin in brain tissue (not mucin). They replaced what I had with a new bottle, but I am seeing the same problems. I believe the formula has been changed to eliminate mercury--don't know if this is the basis of the problems or not--but I believe that is when I started having problems. They have a "new" hematoxylin (560) and eosin (515), but our pathologists did not care for those either. I have been using Surgipath for 14 years and just started having staining problems with it. LuAnn Anderson Neuropthology Lab University of Minnesota At 03:08 PM 11/5/2007, Charles.Embrey wrote: >I find this post most interesting. After using Surgipath Gills III for >the past 7 years we started having a problem with blue mucin just last >month. I corrected the problem by adding GAA but I now have to monitor >the pH on a daily basis. It is strange that after all these years I >would suddenly have this problem and then read about another lab having >the same difficulty with Surgipath Hematoxylin starting about the same >timeframe. Has Surgipath changed something? > >Charles Embrey Jr., PA(ASCP) >Histology Manager, Carle Clinic > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon >Sent: Saturday, November 03, 2007 9:42 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens > >Hello Histonet, > > Over the past several weeks our lab has had problems with >hematoxylin overstaining in the mucin of our gastric specimens. The >degree of hematoxylin overstaining is alarming in the mucin of the >gastric specimens but absent or negligible in other specimens. One >pathologist reviewing the slides indicated some of the overstained >specimens appeared to be inadequately fixed. We have addressed that >issue which showed some improvement, but the problem persists. We are >making changes to increase fixation time (although the specimens now >appear adequately fixed yet retain a large measure of the overstaining), >but any further suggestions for corrective action would be much >appreciated. > > We've been hesitant to increase acid alcohol clearing time because >our other specimens are staining well with the current protocol. > >We are using the following reagents and stain times: > >- Surgipath specimen containers pre-filled with 10% neutral buffered >Formalin. >- Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running >water washes >- .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water >wash >- Surgipath Scott's Tap Water for 1 min followed by running water wash > >Sakura automated stainer set to agitate ( up and down dipping of racks ) >once every 2 seconds, which is the fastest speed available on it. > > >Dick Non >Pathology Department >Ocean County Medical Lab >Brick, New Jersey >_______________________________________________ >Histonet mailing list >Histonet@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: 17 Date: Mon, 5 Nov 2007 16:52:46 -0500 From: "FU,DONTAO" Subject: [Histonet] how to keep thick sections on the slides after deparaffin To: Message-ID: <81FEA2B93DC53C4B8CF3667707CA592A0DCE8C@path-exchange.ad.ufl.edu> Content-Type: text/plain; charset="us-ascii" Hi, I met a big problem in keeping thick sections on the slides after deparaffin recently. I cut a 70um-thick mouse retina section on superfrost plus Gold slides, leaving it dry for 2days. Before deparaffin, I put the slide in the 65C oven for 30 min to let it melt, then I did the general deparaffin procedure. Unfortunately, the section fell off from the slide. Does anyone have any experience on deparaffin thick sections? Any suggestions? Thank you, Ann Dongtao Fu MD Ph.D Lab Manager Molecular Pathology core Dept. of Pathology University of Florida Lab Phone: 352-273-7752 FAX: 352-273-7753 Rm: D11-50 ------------------------------ Message: 18 Date: Mon, 05 Nov 2007 18:51:18 -0500 From: "DNon" Subject: Re: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "Charles.Embrey" Cc: histonet@lists.utsouthwestern.edu Message-ID: <000601c82006$c3081ac0$1f0aa8c0@cyto.ocml.com> Content-Type: text/plain; format=flowed; charset=iso-8859-1; reply-type=original I spoke with Surgipath's QC department today and asked if there was a change in their Gill's III formulation. According to Mike Urban at Surgipath, nothing has changed. Nonetheless, he is going to run additional tests on the lot number in question and will get back to me. I find your post very interesting indeed as our lab has used Surgipath Gill's III for the last 10 years. They ( Surgipath) will also be sending me a gallon of Gill's III from a different lot. If it doesn't show improvement, we'll either lower the ph similar to what you did, or switch to Harris hematoxylin. ----- Original Message ----- From: "Charles.Embrey" To: "DNon" ; Sent: Monday, November 05, 2007 4:08 PM Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Tue, 6 Nov 2007 08:24:24 -0000 From: "Kemlo Rogerson" Subject: RE: [Histonet] how to keep thick sections on the slides after deparaffin To: "FU,DONTAO" , Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F0E6@wahtntex2.waht.swest.nhs.uk> Content-Type: text/plain; charset="us-ascii" Hi, I met a big problem in keeping thick sections on the slides after deparaffin recently. I cut a 70um-thick mouse retina section on superfrost plus Gold slides, leaving it dry for 2days. Before deparaffin, I put the slide in the 65C oven for 30 min to let it melt, then I did the general deparaffin procedure. Unfortunately, the section fell off from the slide. Does anyone have any experience on deparaffin thick sections? Any suggestions? Thank you, You could use bovine albumin to coat the slides (I think 1% but I'm not sure) then dry at 37 degrees for those two days then into the oven at 60 degrees (ish). Think that sort of sets the section onto the slide but you need to get the concentration correct as the protein could stain with some techniques if its too concentrated. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation ------------------------------ Message: 20 Date: Tue, 06 Nov 2007 10:07:04 GMT From: "Barbara Webb" Subject: [Histonet] FYI - Brainbow To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="utf-8" Looking at the fuure? http://www.nytimes.com/2007/11/06/health/research/06brai.html?_r=1&ref=scien ce&oref=slogin ------------------------------ Message: 21 Date: Tue, 6 Nov 2007 05:20:07 -0500 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] Histo, cyto job in NY To: , Message-ID: <000801c8205e$9b559130$0202a8c0@HPPav2> Content-Type: text/plain; charset="us-ascii" If you are looking for a job as a histotech in New York, it's not going to happen. The NY legislators have made a licensure law that only certified med techs can be histotechs. Those already working as histotechs in NY can continue to work in NY. However, any certified histotech moving into NY cannot work as a histotech until they take all the med tech courses and pass the med tech certification exam. NY will not recognize the NAACLS histotech program graduates from SUNY, the only accredited HT program in NY. (SUNY will be closing, as it's impossible to offer all med tech courses and all histotech courses, all in a 2 year program.) NY will not recognize those earning their ASCP HT or HTL certification through the associate degree on-the-job training route. ASCP, NY histotech society, and the NY pathologist groups are lobbying for changes, but the legislators are not listening. NSH has weighed in with ASCP, and the NY pathologists are coming to NSH to see what NSH can do, too. (That's where Vinnie DelaSperanza's (NSH presidents) comments on needing more members come into play. If ASCP and pathologist groups aren't swaying the legislators, then NSH with our fewer numbers are not going to help. Histotechs need to join NSH and ASCP, to histotechs have more numbers and will be listened to better.) The original draft of the bill, according to Vinnie, included histotechs with the appropriate information on training and certification and job responsibilities. The bill got changed over the years, and went unnoticed by NY histotechs. The bill took years (decade+) to pass, so the legislators don't want to change anything on the law at this stage. It may take years to correct. At this time, the med tech programs do not have classes in the theory or practical aspects of histotechnology. I don't know if they are planning on adding them to the med tech curriculum. In the mean time, NY histotechs will be retiring and leaving the field. Workloads will increase as the patient populations ages and technology requires more tissue tests. And there will be fewer and fewer NY histotechs to do these jobs. It's a histology crisis in the making. Peggy A. 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 Xorren@aol.com Sent: Monday, November 05, 2007 1:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histo, cyto job in NY Hello all ! Does anyone know of any histology or cytology prep job in the NY area. I have have 15 years in the field and hope someone has some info. you can respond @ xorren@aol.com or call (516) 671-0907 have a great day !!!!! ************************************** See what's new at http://www.aol.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 22 Date: Tue, 6 Nov 2007 08:21:20 -0500 From: "Pam Barker" Subject: [Histonet] RELIA's Histology Opportunities Update 11-06-07 To: "'Histonet'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi Histonetters! I have been reading some of the posts on the NSH in Denver. What about everybody else, did you make it to the NSH in Denver? If so how was it? What was your favorite thing about it? I havent been since the convention in 2005 in Ft. Lauderdale. Did you have a get together for the histonet? We did at the Ft. Lauderdale convention it was a small turnout but we had fun. I really enjoyed meeting people, learning new things and the party at the Hard Rock Casino. I am planning on attending the 2008 meeting in Pittsburgh. Will I see you there? Here is a list of my current openings all of the positions are full time permanent positions. They are M-F dayshift opportunities and my clients offer excellent compensation, benefits and relocation assistance. And they are ready to interview and hire right away. ****RELIA Spotlight Opportunities**** I am working with a client in the Philadelphia, PA area that has 2 unique and exciting opportunities. These are nonclinical positions. BioSciences Product Manager Histologist Technical and Field Support ********************************* Histology Managers/Supervisors needed in: Dallas, TX Austin, TX Philadelphia, PA Valencia, CA Cape Cod, MA Histotechnicians/Histotechnologists needed in: Phoenix, AZ Atlanta, GA San Francisco, CA Los Angeles, CA Valencia, CA Orlando, FL St. Petersburg, FL Fredericksburg, VA Dallas, TX Waco, TX Austin, TX Philadelphia, PA Pittsburgh, PA If you know of anyone else who might be interested in any of these positions or might like to subscribe to RELIAs Histology Careers Bulletin please feel free to pass this e-mail along to them. Remember it never hurts to look!!! Thanks, Pam 877-60 RELIA (877-607-3542) Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net www.myspace.com/pamatrelia ------------------------------ Message: 23 Date: Tue, 6 Nov 2007 06:41:31 -0800 From: "James Watson" Subject: RE: [Histonet] Histo, cyto job in NY To: , , Message-ID: Content-Type: text/plain; charset="iso-8859-1" At NSH we were told by someone that was in contact with the legistators that wrote the bill that this did not pass and that they were rewriting the bill. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Lee & Peggy Wenk Sent: Tue 11/6/2007 2:20 AM To: Xorren@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histo, cyto job in NY If you are looking for a job as a histotech in New York, it's not going to happen. The NY legislators have made a licensure law that only certified med techs can be histotechs. Those already working as histotechs in NY can continue to work in NY. However, any certified histotech moving into NY cannot work as a histotech until they take all the med tech courses and pass the med tech certification exam. NY will not recognize the NAACLS histotech program graduates from SUNY, the only accredited HT program in NY. (SUNY will be closing, as it's impossible to offer all med tech courses and all histotech courses, all in a 2 year program.) NY will not recognize those earning their ASCP HT or HTL certification through the associate degree on-the-job training route. ASCP, NY histotech society, and the NY pathologist groups are lobbying for changes, but the legislators are not listening. NSH has weighed in with ASCP, and the NY pathologists are coming to NSH to see what NSH can do, too. (That's where Vinnie DelaSperanza's (NSH presidents) comments on needing more members come into play. If ASCP and pathologist groups aren't swaying the legislators, then NSH with our fewer numbers are not going to help. Histotechs need to join NSH and ASCP, to histotechs have more numbers and will be listened to better.) The original draft of the bill, according to Vinnie, included histotechs with the appropriate information on training and certification and job responsibilities. The bill got changed over the years, and went unnoticed by NY histotechs. The bill took years (decade+) to pass, so the legislators don't want to change anything on the law at this stage. It may take years to correct. At this time, the med tech programs do not have classes in the theory or practical aspects of histotechnology. I don't know if they are planning on adding them to the med tech curriculum. In the mean time, NY histotechs will be retiring and leaving the field. Workloads will increase as the patient populations ages and technology requires more tissue tests. And there will be fewer and fewer NY histotechs to do these jobs. It's a histology crisis in the making. Peggy A. 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 Xorren@aol.com Sent: Monday, November 05, 2007 1:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histo, cyto job in NY Hello all ! Does anyone know of any histology or cytology prep job in the NY area. I have have 15 years in the field and hope someone has some info. you can respond @ xorren@aol.com or call (516) 671-0907 have a great day !!!!! ************************************** See what's new at http://www.aol.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 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 6 *************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From b-frederick <@t> northwestern.edu Thu Nov 8 08:17:25 2007 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Thu Nov 8 08:17:42 2007 Subject: [Histonet] TMA arrayers. Does anyone have a favorite model ? In-Reply-To: <0MKwtQ-1IpuWs1nVw-0001gc@mrelayeu.kundenserver.de> Message-ID: <000001c82212$190ce3d0$d00f7ca5@lurie.northwestern.edu> To all, Did anyone see the TMA template (Quick-Ray from Sakura)? It seemed to be kind of neat as well as simple, especially for those 1mm cores. There is a demo video on the website. Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pierre CHAUMAT Sent: Wednesday, November 07, 2007 5:42 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] TMA arrayers. Does anyone have a favorite model ? Dear all, Greg, you mentionned Boekeler but on their web site, I could not find any tissuer arrayer. Would you have a link to an exact page ? Thanks Pierre -----Message d'origine----- De : histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] De la part de histonet-request@lists.utsouthwestern.edu Envoy? : mardi 6 novembre 2007 16:04 ? : histonet@lists.utsouthwestern.edu Objet : Histonet Digest, Vol 48, Issue 6 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. TMA arrayers. Does anyone have a favorite model? (Hugh Luk) 2. Deplasticizing MMA sections (Nancy W. Troiano) 3. Histo, cyto job in NY (Xorren@aol.com) 4. Re: TMA arrayers. Does anyone have a favorite model? (Greg Dobbin) 5. NSH Report (Breeden, Sara) 6. Re: TMA arrayers. Does anyone have a favorite model? (Thomas Pier) 7. Re: Histonet Digest, Vol 48, Issue 5 (Boneslides@aol.com) 8. RE: deplasticizing PMMA sections for staining (Monfils, Paul) 9. Re: TMA arrayers. Web link (Greg Dobbin) 10. NSH show (Rathborne, Toni) 11. RE: Mucin overstained with Hematoxylin in G.I. specimens (Charles.Embrey) 12. RE: Mucin overstained with Hematoxylin in G.I. specimens (Bonnie Whitaker) 13. RE: Mucin overstained with Hematoxylin in G.I. specimens (Douglas D Deltour) 14. RE: deplasticizing PMMA sections for staining (Yaskovich, Ruth A (NIH/NIDCR) [E]) 15. NSH (Bernice Frederick) 16. RE: Mucin overstained with Hematoxylin in G.I. specimens (LuAnn Anderson) 17. how to keep thick sections on the slides after deparaffin (FU,DONTAO) 18. Re: Mucin overstained with Hematoxylin in G.I. specimens (DNon) 19. RE: how to keep thick sections on the slides after deparaffin (Kemlo Rogerson) 20. FYI - Brainbow (Barbara Webb) 21. RE: Histo, cyto job in NY (Lee & Peggy Wenk) 22. RELIA's Histology Opportunities Update 11-06-07 (Pam Barker) 23. RE: Histo, cyto job in NY (James Watson) ---------------------------------------------------------------------- Message: 1 Date: Mon, 5 Nov 2007 08:22:56 -1000 From: Hugh Luk Subject: [Histonet] TMA arrayers. Does anyone have a favorite model? To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct ------------------------------ Message: 2 Date: Mon, 05 Nov 2007 13:25:52 -0500 From: "Nancy W. Troiano" Subject: [Histonet] Deplasticizing MMA sections To: histonet@lists.utsouthwestern.edu Message-ID: <5.2.1.1.2.20071105132352.00c3f668@email.med.yale.edu> Content-Type: text/plain; format=flowed; charset=us-ascii Try soaking the slides in two 5 minute changes of acetone (100%) to deplastify or you can deplastify in 2 changes of Cellosolve (Fisher, cat. #E181-4) 25 minutes each, followed by 5 minutes in 70% ethanol, 40% ethanol, then distilled water for five minutes prior to doing your staining. ------------------------------ Message: 3 Date: Mon, 5 Nov 2007 13:44:13 EST From: Xorren@aol.com Subject: [Histonet] Histo, cyto job in NY To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Hello all ! Does anyone know of any histology or cytology prep job in the NY area. I have have 15 years in the field and hope someone has some info. you can respond @ xorren@aol.com or call (516) 671-0907 have a great day !!!!! ************************************** See what's new at http://www.aol.com ------------------------------ Message: 4 Date: Mon, 05 Nov 2007 14:44:33 -0400 From: "Greg Dobbin" Subject: Re: [Histonet] TMA arrayers. Does anyone have a favorite model? To: Message-ID: Content-Type: text/plain; charset=US-ASCII I am investigating an instrument that I saw at NSH by RMC Products. Model no. is KIN-1. The picture on the web site does not match exactly the unit they had at NSH but they could send you the same broshure they were passing out at their booth (it must be available as a pdf). Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> hlukey@msn.com 11/5/2007 2:22 PM >>> I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct__ _____________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. ------------------------------ Message: 5 Date: Mon, 5 Nov 2007 11:50:55 -0700 From: "Breeden, Sara" Subject: [Histonet] NSH Report To: Message-ID: <4D14F0FC9316DD41972D5F03C070908B8F4725@nmdamailsvr.nmda.ad.nmsu.edu> Content-Type: text/plain; charset="us-ascii" Okay, I'm back from my week at NSH and this is my report. 1. The NSH staff is awesome. Aubrey, Carrie and the entire NSH staff pulled off yet another flawless meeting. We can't thank NSH enough for all they do! 2. Joe Nocito is alive and well. I met him. He lives and he wasn't wearing a flak jacket. 3. Our vendors are magnificent - supportive, creative, helpful and approachable. Kudos!! 4. Each one of us needs to recruit NSH members - there is strength in numbers. NSH is our connection in SO many ways! 5. Ada Feldman makes a terrific clown! 6. Did you know that histologists are the ONLY technicians not recognized by the US Government as healthcare professionals? 7. After 3 days of walking the 6 blocks from hotel to convention center, I discovered the free bus. 8. There must be a "message board" at next year's meeting so we can find each other if we need to. 9. I want to know what vitamins Peggy Wenk takes. Seriously. Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 ------------------------------ Message: 6 Date: Mon, 05 Nov 2007 12:55:36 -0600 From: "Thomas Pier" Subject: Re: [Histonet] TMA arrayers. Does anyone have a favorite model? To: , Message-ID: <472F12C8020000DF00002B3D@gwmail.medicine.wisc.edu> Content-Type: text/plain; charset=US-ASCII I haven't used any other instruments, but I've been happy with the Beecher MTA-1. Tom Pier >>> Hugh Luk 11/05/07 12:22 PM >>> I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct__ _____________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Mon, 5 Nov 2007 14:10:40 EST From: Boneslides@aol.com Subject: [Histonet] Re: Histonet Digest, Vol 48, Issue 5 To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" I de-plasticize my PMMA sections in methyl acetate...takes about 30 minutes, then rehydrate thru several changes of absolute, 95% and 70% alcohol, rinse in water, stain as desired. Diane M. Mahovlic, HT, MLT(ASCP) Orthopedic Pathology & Biomaterials Laboratory Department of Anatomic Pathology The Cleveland Clinic Foundation 9500 Euclid Avenue- L30 Cleveland, Ohio 44195 216-444-0166 ************************************** See what's new at http://www.aol.com ------------------------------ Message: 8 Date: Mon, 5 Nov 2007 14:22:08 -0500 From: "Monfils, Paul" Subject: RE: [Histonet] deplasticizing PMMA sections for staining To: Message-ID: <4EBFF65383B74D49995298C4976D1D5E273CF6@LSRIEXCH1.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" In my experience, xylene is a rather poor solvent for PMMA. It works much faster at 60 degrees, but don't try heating flammable solvents in your oven unless you are certain it is certified explosion-proof (most paraffin ovens are not). Even then, unless you have the oven in or at least next to a fume hood, you'll get a lot of xylene vapors in the air as a result of heating it. I deplasticise PMMA with 2-methoxyethyl acetate, which only takes a couple of hours at room temperature. This has a rather strong though not unpleasant odor, and should be used in the hood. ------------------------------ Message: 9 Date: Mon, 05 Nov 2007 16:12:32 -0400 From: "Greg Dobbin" Subject: Re: [Histonet] TMA arrayers. Web link To: Message-ID: Content-Type: text/plain; charset=US-ASCII Sorry. I see the company name is actually Boeckeler and the website is www.boeckeler.com Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> "Greg Dobbin" 11/5/2007 2:44 PM >>> I am investigating an instrument that I saw at NSH by RMC Products. Model no. is KIN-1. The picture on the web site does not match exactly the unit they had at NSH but they could send you the same broshure they were passing out at their booth (it must be available as a pdf). Cheers! Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> hlukey@msn.com 11/5/2007 2:22 PM >>> I need advice. Is there a favorite Tissue microarray unit? I currently use the Beecher TMA #1, but due to the humidity in Hawaii, the electronics have failed. I guess I shouldn't have taken it swimming with me in the ocean! I attending NSH, but did not see too many TMA vendors. Beecher did throw a very nice break-time pretzel party, but I missed meeting any of their people. I would appreciate any advice... Thanks, Hugh Luk, HTL (ASCP) hlukey@msn.com or hluk@crch.hawaii.edu Pathology Shared Resources Lab Cancer Research Center of Hawaii Honolulu, Hawaii _________________________________________________________________ Climb to the top of the charts! Play Star Shuffle: the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct__ _____________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. ------------------------------ Message: 10 Date: Mon, 5 Nov 2007 15:30:49 -0500 From: "Rathborne, Toni" Subject: [Histonet] NSH show To: Message-ID: Content-Type: text/plain; charset="utf-8" How was the show? I heard there were a lot of vendors there. What about the lectures/workshops? I was hoping to go, so many relevant topics offered. A friend of mine was there and mentioned that Ventana had some sort of secretive booth. What was that all about? Is it a new IHC stainer? Or part of their whole "lab platform"? Toni CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. Somerset Medical Center is proud to receive the Somerset County Business Partnership's 2006 Quality of Life Award. ------------------------------ Message: 11 Date: Mon, 5 Nov 2007 15:08:54 -0600 From: "Charles.Embrey" Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "DNon" , Message-ID: <44780C571F28624DBB446DE55C4D733A1FE565@EXCHANGEBE1.carle.com> Content-Type: text/plain; charset="us-ascii" I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Mon, 5 Nov 2007 15:22:45 -0600 From: "Bonnie Whitaker" Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "'Charles.Embrey'" , "'DNon'" , Message-ID: <003501c81ff2$02602060$3601a8c0@brownpathology.net> Content-Type: text/plain; charset="us-ascii" Hi All, I had the same type incident (with the increase in mucin staining with the Surgipath hematoxylin), but it was a one time thing that we attributed to a larger than normal volume of H&E's in that particular time period. We increased the frequency that solutions are changed slightly, and haven't had another problem. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, November 05, 2007 3:09 PM To: DNon; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@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: 13 Date: Mon, 5 Nov 2007 16:26:45 -0500 From: "Douglas D Deltour" Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "'Charles.Embrey'" , "'DNon'" , Message-ID: Content-Type: text/plain; charset="US-ASCII" Charles, We also are having the same issue with the Surgipath Gills III. This issue also started last month. Very interesting. Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, November 05, 2007 4:09 PM To: DNon; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@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: 14 Date: Mon, 5 Nov 2007 16:43:34 -0500 From: "Yaskovich, Ruth A (NIH/NIDCR) [E]" Subject: RE: [Histonet] deplasticizing PMMA sections for staining To: "John Baker" , Message-ID: Content-Type: text/plain; charset="us-ascii" John, You can soak them in 3 changes of acetone for 15 minutes each. Put them in the first acetone right out of the oven. Ruth Yaskovich National Institutes of Health National Institute of Dental and Craniofacial Research -----Original Message----- From: John Baker [mailto:bakerj@umich.edu] Sent: Monday, November 05, 2007 12:07 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] deplasticizing PMMA sections for staining > > > Hello All, We are sectioning PMMA embedded rat femur at 5 microns on > the Polycut. Then we are deplasticizing for H & E and Trichrome > stains. It has been taking over 2 weeks with several changes of xylene > to get them done and still some remnants of plastic there. I see in > Sheehan's Practice and Theory of Histotechnology that in several > protocols they deplasticize in xylene for 4 hours at 55 degrees C. > The flash point of xylene is 25 degree C. Is it safe in covered > staining dishes to actually do this? Also, we use Mayer's Hematoxylin > and 2% Eosin to stain. What H & E protocol do you use for bone? Any > suggestions on how to speed up the deplasticizing process welcome. > Does anyone have Diane Sterchi's and Gayle Callis's email addresses, > lost them? > Thanks, John > > > > > John A. Baker The University of Michigan Orthopaedic Research Laboratories Histology Unit 109 Zina Pitcher Place, 2218 BSRB Ann Arbor, MI 48109-2200 734-936-1635 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Mon, 5 Nov 2007 15:45:56 -0600 From: "Bernice Frederick" Subject: [Histonet] NSH To: Message-ID: <000001c81ff5$42a1aec0$d00f7ca5@lurie.northwestern.edu> Content-Type: text/plain; charset="us-ascii" This was my first meeting and whereas I enjoyed myself tremendously I only had 2 gripes 1. There should be a block of time set aside to see the vendors. Most of us were in workshops for a lot of the time and had to do quick run up to the vendors. 2. I was surprised water was only provided for the speakers. It was so dry I was going through 36-48 oz of water every day and at 2$ a pop... If I'd known, I'd have run off to Walgreens or somewhere. I do plan to be back and really enjoyed everybody and everything. Missed meeting Joe though I thought we had arranged for that (tee hee). Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 ------------------------------ Message: 16 Date: Mon, 05 Nov 2007 15:51:47 -0600 From: LuAnn Anderson Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "Charles.Embrey" , "DNon" , Message-ID: Content-Type: text/plain; charset="us-ascii"; format=flowed I too have had problems recently with Surgipath hematoxylin in brain tissue (not mucin). They replaced what I had with a new bottle, but I am seeing the same problems. I believe the formula has been changed to eliminate mercury--don't know if this is the basis of the problems or not--but I believe that is when I started having problems. They have a "new" hematoxylin (560) and eosin (515), but our pathologists did not care for those either. I have been using Surgipath for 14 years and just started having staining problems with it. LuAnn Anderson Neuropthology Lab University of Minnesota At 03:08 PM 11/5/2007, Charles.Embrey wrote: >I find this post most interesting. After using Surgipath Gills III for >the past 7 years we started having a problem with blue mucin just last >month. I corrected the problem by adding GAA but I now have to monitor >the pH on a daily basis. It is strange that after all these years I >would suddenly have this problem and then read about another lab having >the same difficulty with Surgipath Hematoxylin starting about the same >timeframe. Has Surgipath changed something? > >Charles Embrey Jr., PA(ASCP) >Histology Manager, Carle Clinic > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon >Sent: Saturday, November 03, 2007 9:42 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens > >Hello Histonet, > > Over the past several weeks our lab has had problems with >hematoxylin overstaining in the mucin of our gastric specimens. The >degree of hematoxylin overstaining is alarming in the mucin of the >gastric specimens but absent or negligible in other specimens. One >pathologist reviewing the slides indicated some of the overstained >specimens appeared to be inadequately fixed. We have addressed that >issue which showed some improvement, but the problem persists. We are >making changes to increase fixation time (although the specimens now >appear adequately fixed yet retain a large measure of the overstaining), >but any further suggestions for corrective action would be much >appreciated. > > We've been hesitant to increase acid alcohol clearing time because >our other specimens are staining well with the current protocol. > >We are using the following reagents and stain times: > >- Surgipath specimen containers pre-filled with 10% neutral buffered >Formalin. >- Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running >water washes >- .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water >wash >- Surgipath Scott's Tap Water for 1 min followed by running water wash > >Sakura automated stainer set to agitate ( up and down dipping of racks ) >once every 2 seconds, which is the fastest speed available on it. > > >Dick Non >Pathology Department >Ocean County Medical Lab >Brick, New Jersey >_______________________________________________ >Histonet mailing list >Histonet@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: 17 Date: Mon, 5 Nov 2007 16:52:46 -0500 From: "FU,DONTAO" Subject: [Histonet] how to keep thick sections on the slides after deparaffin To: Message-ID: <81FEA2B93DC53C4B8CF3667707CA592A0DCE8C@path-exchange.ad.ufl.edu> Content-Type: text/plain; charset="us-ascii" Hi, I met a big problem in keeping thick sections on the slides after deparaffin recently. I cut a 70um-thick mouse retina section on superfrost plus Gold slides, leaving it dry for 2days. Before deparaffin, I put the slide in the 65C oven for 30 min to let it melt, then I did the general deparaffin procedure. Unfortunately, the section fell off from the slide. Does anyone have any experience on deparaffin thick sections? Any suggestions? Thank you, Ann Dongtao Fu MD Ph.D Lab Manager Molecular Pathology core Dept. of Pathology University of Florida Lab Phone: 352-273-7752 FAX: 352-273-7753 Rm: D11-50 ------------------------------ Message: 18 Date: Mon, 05 Nov 2007 18:51:18 -0500 From: "DNon" Subject: Re: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens To: "Charles.Embrey" Cc: histonet@lists.utsouthwestern.edu Message-ID: <000601c82006$c3081ac0$1f0aa8c0@cyto.ocml.com> Content-Type: text/plain; format=flowed; charset=iso-8859-1; reply-type=original I spoke with Surgipath's QC department today and asked if there was a change in their Gill's III formulation. According to Mike Urban at Surgipath, nothing has changed. Nonetheless, he is going to run additional tests on the lot number in question and will get back to me. I find your post very interesting indeed as our lab has used Surgipath Gill's III for the last 10 years. They ( Surgipath) will also be sending me a gallon of Gill's III from a different lot. If it doesn't show improvement, we'll either lower the ph similar to what you did, or switch to Harris hematoxylin. ----- Original Message ----- From: "Charles.Embrey" To: "DNon" ; Sent: Monday, November 05, 2007 4:08 PM Subject: RE: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens I find this post most interesting. After using Surgipath Gills III for the past 7 years we started having a problem with blue mucin just last month. I corrected the problem by adding GAA but I now have to monitor the pH on a daily basis. It is strange that after all these years I would suddenly have this problem and then read about another lab having the same difficulty with Surgipath Hematoxylin starting about the same timeframe. Has Surgipath changed something? Charles Embrey Jr., PA(ASCP) Histology Manager, Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DNon Sent: Saturday, November 03, 2007 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mucin overstained with Hematoxylin in G.I. specimens Hello Histonet, Over the past several weeks our lab has had problems with hematoxylin overstaining in the mucin of our gastric specimens. The degree of hematoxylin overstaining is alarming in the mucin of the gastric specimens but absent or negligible in other specimens. One pathologist reviewing the slides indicated some of the overstained specimens appeared to be inadequately fixed. We have addressed that issue which showed some improvement, but the problem persists. We are making changes to increase fixation time (although the specimens now appear adequately fixed yet retain a large measure of the overstaining), but any further suggestions for corrective action would be much appreciated. We've been hesitant to increase acid alcohol clearing time because our other specimens are staining well with the current protocol. We are using the following reagents and stain times: - Surgipath specimen containers pre-filled with 10% neutral buffered Formalin. - Surgipath Gills III Hematoxlyin - 6.5 minutes followed by running water washes - .75% HCL in 70% ETOH for 1 second (1 dip) followed by running water wash - Surgipath Scott's Tap Water for 1 min followed by running water wash Sakura automated stainer set to agitate ( up and down dipping of racks ) once every 2 seconds, which is the fastest speed available on it. Dick Non Pathology Department Ocean County Medical Lab Brick, New Jersey _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Tue, 6 Nov 2007 08:24:24 -0000 From: "Kemlo Rogerson" Subject: RE: [Histonet] how to keep thick sections on the slides after deparaffin To: "FU,DONTAO" , Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F0E6@wahtntex2.waht.swest.nhs.uk> Content-Type: text/plain; charset="us-ascii" Hi, I met a big problem in keeping thick sections on the slides after deparaffin recently. I cut a 70um-thick mouse retina section on superfrost plus Gold slides, leaving it dry for 2days. Before deparaffin, I put the slide in the 65C oven for 30 min to let it melt, then I did the general deparaffin procedure. Unfortunately, the section fell off from the slide. Does anyone have any experience on deparaffin thick sections? Any suggestions? Thank you, You could use bovine albumin to coat the slides (I think 1% but I'm not sure) then dry at 37 degrees for those two days then into the oven at 60 degrees (ish). Think that sort of sets the section onto the slide but you need to get the concentration correct as the protein could stain with some techniques if its too concentrated. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation ------------------------------ Message: 20 Date: Tue, 06 Nov 2007 10:07:04 GMT From: "Barbara Webb" Subject: [Histonet] FYI - Brainbow To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="utf-8" Looking at the fuure? http://www.nytimes.com/2007/11/06/health/research/06brai.html?_r=1&ref=scien ce&oref=slogin ------------------------------ Message: 21 Date: Tue, 6 Nov 2007 05:20:07 -0500 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] Histo, cyto job in NY To: , Message-ID: <000801c8205e$9b559130$0202a8c0@HPPav2> Content-Type: text/plain; charset="us-ascii" If you are looking for a job as a histotech in New York, it's not going to happen. The NY legislators have made a licensure law that only certified med techs can be histotechs. Those already working as histotechs in NY can continue to work in NY. However, any certified histotech moving into NY cannot work as a histotech until they take all the med tech courses and pass the med tech certification exam. NY will not recognize the NAACLS histotech program graduates from SUNY, the only accredited HT program in NY. (SUNY will be closing, as it's impossible to offer all med tech courses and all histotech courses, all in a 2 year program.) NY will not recognize those earning their ASCP HT or HTL certification through the associate degree on-the-job training route. ASCP, NY histotech society, and the NY pathologist groups are lobbying for changes, but the legislators are not listening. NSH has weighed in with ASCP, and the NY pathologists are coming to NSH to see what NSH can do, too. (That's where Vinnie DelaSperanza's (NSH presidents) comments on needing more members come into play. If ASCP and pathologist groups aren't swaying the legislators, then NSH with our fewer numbers are not going to help. Histotechs need to join NSH and ASCP, to histotechs have more numbers and will be listened to better.) The original draft of the bill, according to Vinnie, included histotechs with the appropriate information on training and certification and job responsibilities. The bill got changed over the years, and went unnoticed by NY histotechs. The bill took years (decade+) to pass, so the legislators don't want to change anything on the law at this stage. It may take years to correct. At this time, the med tech programs do not have classes in the theory or practical aspects of histotechnology. I don't know if they are planning on adding them to the med tech curriculum. In the mean time, NY histotechs will be retiring and leaving the field. Workloads will increase as the patient populations ages and technology requires more tissue tests. And there will be fewer and fewer NY histotechs to do these jobs. It's a histology crisis in the making. Peggy A. 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 Xorren@aol.com Sent: Monday, November 05, 2007 1:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histo, cyto job in NY Hello all ! Does anyone know of any histology or cytology prep job in the NY area. I have have 15 years in the field and hope someone has some info. you can respond @ xorren@aol.com or call (516) 671-0907 have a great day !!!!! ************************************** See what's new at http://www.aol.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 22 Date: Tue, 6 Nov 2007 08:21:20 -0500 From: "Pam Barker" Subject: [Histonet] RELIA's Histology Opportunities Update 11-06-07 To: "'Histonet'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi Histonetters! I have been reading some of the posts on the NSH in Denver. What about everybody else, did you make it to the NSH in Denver? If so how was it? What was your favorite thing about it? I havent been since the convention in 2005 in Ft. Lauderdale. Did you have a get together for the histonet? We did at the Ft. Lauderdale convention it was a small turnout but we had fun. I really enjoyed meeting people, learning new things and the party at the Hard Rock Casino. I am planning on attending the 2008 meeting in Pittsburgh. Will I see you there? Here is a list of my current openings all of the positions are full time permanent positions. They are M-F dayshift opportunities and my clients offer excellent compensation, benefits and relocation assistance. And they are ready to interview and hire right away. ****RELIA Spotlight Opportunities**** I am working with a client in the Philadelphia, PA area that has 2 unique and exciting opportunities. These are nonclinical positions. BioSciences Product Manager Histologist Technical and Field Support ********************************* Histology Managers/Supervisors needed in: Dallas, TX Austin, TX Philadelphia, PA Valencia, CA Cape Cod, MA Histotechnicians/Histotechnologists needed in: Phoenix, AZ Atlanta, GA San Francisco, CA Los Angeles, CA Valencia, CA Orlando, FL St. Petersburg, FL Fredericksburg, VA Dallas, TX Waco, TX Austin, TX Philadelphia, PA Pittsburgh, PA If you know of anyone else who might be interested in any of these positions or might like to subscribe to RELIAs Histology Careers Bulletin please feel free to pass this e-mail along to them. Remember it never hurts to look!!! Thanks, Pam 877-60 RELIA (877-607-3542) Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net www.myspace.com/pamatrelia ------------------------------ Message: 23 Date: Tue, 6 Nov 2007 06:41:31 -0800 From: "James Watson" Subject: RE: [Histonet] Histo, cyto job in NY To: , , Message-ID: Content-Type: text/plain; charset="iso-8859-1" At NSH we were told by someone that was in contact with the legistators that wrote the bill that this did not pass and that they were rewriting the bill. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Lee & Peggy Wenk Sent: Tue 11/6/2007 2:20 AM To: Xorren@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histo, cyto job in NY If you are looking for a job as a histotech in New York, it's not going to happen. The NY legislators have made a licensure law that only certified med techs can be histotechs. Those already working as histotechs in NY can continue to work in NY. However, any certified histotech moving into NY cannot work as a histotech until they take all the med tech courses and pass the med tech certification exam. NY will not recognize the NAACLS histotech program graduates from SUNY, the only accredited HT program in NY. (SUNY will be closing, as it's impossible to offer all med tech courses and all histotech courses, all in a 2 year program.) NY will not recognize those earning their ASCP HT or HTL certification through the associate degree on-the-job training route. ASCP, NY histotech society, and the NY pathologist groups are lobbying for changes, but the legislators are not listening. NSH has weighed in with ASCP, and the NY pathologists are coming to NSH to see what NSH can do, too. (That's where Vinnie DelaSperanza's (NSH presidents) comments on needing more members come into play. If ASCP and pathologist groups aren't swaying the legislators, then NSH with our fewer numbers are not going to help. Histotechs need to join NSH and ASCP, to histotechs have more numbers and will be listened to better.) The original draft of the bill, according to Vinnie, included histotechs with the appropriate information on training and certification and job responsibilities. The bill got changed over the years, and went unnoticed by NY histotechs. The bill took years (decade+) to pass, so the legislators don't want to change anything on the law at this stage. It may take years to correct. At this time, the med tech programs do not have classes in the theory or practical aspects of histotechnology. I don't know if they are planning on adding them to the med tech curriculum. In the mean time, NY histotechs will be retiring and leaving the field. Workloads will increase as the patient populations ages and technology requires more tissue tests. And there will be fewer and fewer NY histotechs to do these jobs. It's a histology crisis in the making. Peggy A. 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 Xorren@aol.com Sent: Monday, November 05, 2007 1:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histo, cyto job in NY Hello all ! Does anyone know of any histology or cytology prep job in the NY area. I have have 15 years in the field and hope someone has some info. you can respond @ xorren@aol.com or call (516) 671-0907 have a great day !!!!! ************************************** See what's new at http://www.aol.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 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 6 *************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hej01 <@t> health.state.ny.us Thu Nov 8 08:22:44 2007 From: hej01 <@t> health.state.ny.us (Helen E Johnson) Date: Thu Nov 8 08:23:05 2007 Subject: [Histonet] mouse histology atlas Message-ID: Hi Histonetters, Does anyone know of a good mouse histology atlas? Helen Johnson (hej01@health.state.ny.us) IMPORTANT NOTICE: This e-mail and any attachments may contain confidential or sensitive information which is, or may be, legally privileged or otherwise protected by law from further disclosure. It is intended only for the addressee. If you received this in error or from someone who was not authorized to send it to you, please do not distribute, copy or use it or any attachments. Please notify the sender immediately by reply e-mail and delete this from your system. Thank you for your cooperation. From arnie.jimenez <@t> vel-lab.com Thu Nov 8 08:23:20 2007 From: arnie.jimenez <@t> vel-lab.com (arnie.jimenez@vel-lab.com) Date: Thu Nov 8 08:25:33 2007 Subject: [Histonet] Re: Away from Office Message-ID: <20071108142320.15276.qmail@plesk5.hostgator.com> Thank you for contacting Vel-Lab Research. We will be out of the lab starting Nov. 7th and will return on Nov. 12th. We will check e-mail regularly but will be unable to respond. Please do not mail any packages to our lab until Nov. 12th. Regards, Arnie Jimenez Vel-Lab Research From gvdobbin <@t> ihis.org Thu Nov 8 08:59:21 2007 From: gvdobbin <@t> ihis.org (Greg Dobbin) Date: Thu Nov 8 08:59:46 2007 Subject: [Histonet] Short processing schedule-suggestions Message-ID: Hello All, I would like to get your suggestions for a processing schedule for small biopsies (cores, GI's, other endoscopics). I want to process these little guys separate from everything else to hopefully improve section quality and perhaps staining quality. I don't want the absolute shortest possible schedule. We have a newer VIP5 for our main processor (doing everything on one schedule currently) and a much older VIP sitting idle as a backup that I plan to start using for the biopsies. We process overnight so I have more than enough time. I will just delay the biopsy processing so that both processors are finished at essentially the same time. What I need is a shorter schedule for the biopsies so that the detrimental affects of xylene and alcohol are minimized (and yes, we still use xylene! And in case it matters schedule-wise, we use 10% NBFS as well). Thanks in advance. Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. From RSRICHMOND <@t> aol.com Thu Nov 8 10:08:27 2007 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Thu Nov 8 10:08:52 2007 Subject: [Histonet] Re: Formaline-free fixative Message-ID: Pierre Chaumat (where are you, Pierre?) asks: >>Has anyone tested alternative solution to formaline fixation ? Has anyone switched to a new tissue fixative ? - I would like to share some experience.<< About the only alternative "fixative" that actually is a fixative is glyoxal, available under numerous trade names. Its fixative properties are somewhat different from those of formaldehyde. In particular, immunostains may require adjustment, and comparison with formaldehyde fixed tissue from the same case. In the USA, the breast cancer immunostains are required by the federal govenrment to be done on formaldehyde fixed tissue. Do not use any trade-named fixative whose composition is secret. Sometimes fairly adequate information can be obtained from the Materials Safety Data Sheet (MSDS), in the USA. Does France require similar documentation of hazardous materials? If so, what is this document called, and what is a good Web site to look some of them up? (I can read French.) Bob Richmond Samurai Pathologist Knoxville, Tennessee From mauger <@t> email.chop.edu Thu Nov 8 10:50:43 2007 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Thu Nov 8 10:51:24 2007 Subject: [Histonet] B5 substitute Message-ID: Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger From rjbuesa <@t> yahoo.com Thu Nov 8 10:59:40 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 8 10:59:45 2007 Subject: [Histonet] Artefacts In-Reply-To: <86ADE4EB583CE64799A9924684A0FBBF0222F121@wahtntex2.waht.swest.nhs.uk> Message-ID: <222219.92551.qm@web61219.mail.yahoo.com> and most likely the first! Ren? J. Kemlo Rogerson wrote: Ah the spotty H&E problem again. 1) Inadequate dewaxing. 2) Inadequate fixation. 3) Inadequate processing. Not necessarily in that order. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation _______________________________________________ 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 rjbuesa <@t> yahoo.com Thu Nov 8 11:03:38 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 8 11:03:43 2007 Subject: [Histonet] Short processing schedule-suggestions In-Reply-To: Message-ID: <462982.42437.qm@web61214.mail.yahoo.com> >From my experience in processing, the most important thing is to maintain dehydration time to clearing time to infiltration time in constant proportions once you find a good protocol. If you "long" protocol works well for you, reduce the time in each step in a way that at the end will be completed in the time you need, BUT preserving the time proportions you have now. Ren? J. Greg Dobbin wrote: Hello All, I would like to get your suggestions for a processing schedule for small biopsies (cores, GI's, other endoscopics). I want to process these little guys separate from everything else to hopefully improve section quality and perhaps staining quality. I don't want the absolute shortest possible schedule. We have a newer VIP5 for our main processor (doing everything on one schedule currently) and a much older VIP sitting idle as a backup that I plan to start using for the biopsies. We process overnight so I have more than enough time. I will just delay the biopsy processing so that both processors are finished at essentially the same time. What I need is a shorter schedule for the biopsies so that the detrimental affects of xylene and alcohol are minimized (and yes, we still use xylene! And in case it matters schedule-wise, we use 10% NBFS as well). Thanks in advance. Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. _______________________________________________ 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 SDrew <@t> uwhealth.org Thu Nov 8 11:07:25 2007 From: SDrew <@t> uwhealth.org (Drew Sally A.) Date: Thu Nov 8 11:08:52 2007 Subject: [Histonet] Clone for Prog. Receptor Message-ID: We currently are using the 1A6 clone for our progesterone receptor(PR) IHC. We were wondering how other people felt about rabbit monoclonal PR antibodies. Does anyone have strong preferences or experiences with other PR clones-mouse or rabbit-that they'd like to share? Thank you...! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 From jkiernan <@t> uwo.ca Thu Nov 8 11:10:47 2007 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Thu Nov 8 11:10:51 2007 Subject: [Histonet] Artefacts Message-ID: Dear Toxicology You need to explain your problem more clearly. Please include histonet@lists.utsouthwestern.edu in your replies. We all want to share our knowledge. What tissue are you staining? How was it fixed? Are you staining dewaxed and hydrated paraffin sections? Cryostat sections? Which haemalum & eosin method did you use? Does your lab have a book with instructions for H&E staining? If not, why not? Are the "faint stained spots on sections" blue, purple, pink or red? Where are these spots? John Kiernan Anatomy, UWO London, Canada. === ----- Original Message ----- From: Toxicology Date: Thursday, November 8, 2007 1:36 Subject: [Histonet] Artefacts To: histonet@lists.utsouthwestern.edu > Dear all, > I am facing a problem in Hematoxylin and Eosin staining. There > are several > faint stained spots on sections and because of that its > difficuly to > interpret the lesions. I am unable to sort out the > problem. Can any one > help me? > > > > > -------------------------Email Disclaimer------------------------ > --------- > From Terry.Marshall <@t> rothgen.nhs.uk Thu Nov 8 11:13:21 2007 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Thu Nov 8 11:13:29 2007 Subject: [Histonet] Artefacts Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE4F@TRFT-EX01.xRothGen.nhs.uk> Some of these questions should be answered by the photos, but I cannot find them. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: 08 November 2007 17:11 To: Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Artefacts Dear Toxicology You need to explain your problem more clearly. Please include histonet@lists.utsouthwestern.edu in your replies. We all want to share our knowledge. What tissue are you staining? How was it fixed? Are you staining dewaxed and hydrated paraffin sections? Cryostat sections? Which haemalum & eosin method did you use? Does your lab have a book with instructions for H&E staining? If not, why not? Are the "faint stained spots on sections" blue, purple, pink or red? Where are these spots? John Kiernan Anatomy, UWO London, Canada. === ----- Original Message ----- From: Toxicology Date: Thursday, November 8, 2007 1:36 Subject: [Histonet] Artefacts To: histonet@lists.utsouthwestern.edu > Dear all, > I am facing a problem in Hematoxylin and Eosin staining. There are > several faint stained spots on sections and because of that its > difficuly to interpret the lesions. I am unable to sort out the > problem. Can any one help me? > > > > > -------------------------Email Disclaimer------------------------ > --------- > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From fudo <@t> ufl.edu Thu Nov 8 11:13:22 2007 From: fudo <@t> ufl.edu (FU,DONGTAO) Date: Thu Nov 8 11:13:36 2007 Subject: [Histonet] problem in dual staining of CD4 and FOXP3 Message-ID: <813060655.505211194542002919.JavaMail.osg@osgjas04.cns.ufl.edu> Hello, Can anyone give me some suggestions on my case? I know a lot of people here have a lot of dual staining experiences. I did dual staining on 2 antibodies from different species in the past. They worked well. But using 2 antibodies from same species is my first time try and I met a big problem. Any suggestions I will be very appreciate. Ann On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" wrote: > Hi, all > > Thank you first for giving me some good suggestions on > thick-section question I posted last time. Now I met another > problem when I did CD4 and FOXP3 dual staining using murine fresh > frozen spleen. If I did single staining, both antibodies worked > very well. However if I did dual staining, I could only get good > result from the first antibody. The second one did not work at > all(I mean no specific staining). I used CD4 from BD(rat > anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. > I think there might be something wrong with my serum block(I used > normal rat serum block) before I added secondary antibody. Or any > other serum block I need to add to decrease the non-specific > binding which I have not done yet. Does anyone can give me some > suggestions according to my protocol below? How can I get > specific staining of the secondary (primary)antibody? Thank you, > > Below is the simple protocol I used for dual staining: > 1. 2% normal goat serum block 20 min > 2. 1* antibody CD4 1:750 in diluent O/N 4C > 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT > 4. Serum block: 5% normal rat serum 30 min > 5. 1* antibody FOXP3 1:100 in diluent 1h RT > 6. Secondary AF488 Donkey anti-rat in TBS 1h RT > > Use 1xTBS as wash buffer. Before staining, fix tissue in -20C > Aceton for 5 min, then airdry. > > > Ann Dongtao Fu MD Ph.D > Lab Manager > Molecular Pathology core > Dept. of Pathology > University of Florida > 1600 SW Archer Road > Gainesville, FL 32610 > Lab Phone: 352-273-7752 > FAX: 352-273-7753 > Rm: D11-50 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Ann Dongtao Fu MD, Ph.D Lab Manager Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 From JWEEMS <@t> sjha.org Thu Nov 8 11:16:04 2007 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Thu Nov 8 11:16:24 2007 Subject: [Histonet] Artefacts In-Reply-To: <5C0BED61F529364E86309CADEA63FEF2F3AE4F@TRFT-EX01.xRothGen.nhs.uk> Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD32048F41AF@sjhaexc02.sjha.org> The web site says give it 24 hrs for the photo to post... Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, November 08, 2007 12:13 PM To: John Kiernan; Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Artefacts Some of these questions should be answered by the photos, but I cannot find them. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: 08 November 2007 17:11 To: Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Artefacts Dear Toxicology You need to explain your problem more clearly. Please include histonet@lists.utsouthwestern.edu in your replies. We all want to share our knowledge. What tissue are you staining? How was it fixed? Are you staining dewaxed and hydrated paraffin sections? Cryostat sections? Which haemalum & eosin method did you use? Does your lab have a book with instructions for H&E staining? If not, why not? Are the "faint stained spots on sections" blue, purple, pink or red? Where are these spots? John Kiernan Anatomy, UWO London, Canada. === ----- Original Message ----- From: Toxicology Date: Thursday, November 8, 2007 1:36 Subject: [Histonet] Artefacts To: histonet@lists.utsouthwestern.edu > Dear all, > I am facing a problem in Hematoxylin and Eosin staining. There are > several faint stained spots on sections and because of that its > difficuly to interpret the lesions. I am unable to sort out the > problem. Can any one help me? > > > > > -------------------------Email Disclaimer------------------------ > --------- > _______________________________________________ Histonet mailing list Histonet@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 Joseph's Health System, Inc. From JWEEMS <@t> sjha.org Thu Nov 8 11:20:12 2007 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Thu Nov 8 11:20:27 2007 Subject: [Histonet] B5 substitute In-Reply-To: Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD32048F41B0@sjhaexc02.sjha.org> We use B-Plus from BBC. http://www.bbcus.com/website_search.html It takes a long time to get used to, especially when they have to compare to previous mercury fixed cases, but it can be done! Good luck, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne Mauger Sent: Thursday, November 08, 2007 11:51 AM To: gvdobbin@ihis.org; Histonet@lists.utsouthwestern.edu Subject: [Histonet] B5 substitute Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ 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 Joseph's Health System, Inc. From mohs76009 <@t> yahoo.com Thu Nov 8 11:24:17 2007 From: mohs76009 <@t> yahoo.com (Matt Bancroft) Date: Thu Nov 8 11:24:21 2007 Subject: [Histonet] B5 substitute In-Reply-To: Message-ID: <704342.79971.qm@web63407.mail.re1.yahoo.com> B-Plus Joanne Mauger wrote: Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ 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 Terry.Marshall <@t> rothgen.nhs.uk Thu Nov 8 11:27:01 2007 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Thu Nov 8 11:27:08 2007 Subject: [Histonet] Artefacts Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE50@TRFT-EX01.xRothGen.nhs.uk> Yes, I know. Posters can read that too and wait until the photos appear before they post the question. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: 08 November 2007 17:16 To: Marshall Terry Dr, Consultant Histopathologist; John Kiernan; Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Artefacts The web site says give it 24 hrs for the photo to post... Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, November 08, 2007 12:13 PM To: John Kiernan; Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Artefacts Some of these questions should be answered by the photos, but I cannot find them. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: 08 November 2007 17:11 To: Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Artefacts Dear Toxicology You need to explain your problem more clearly. Please include histonet@lists.utsouthwestern.edu in your replies. We all want to share our knowledge. What tissue are you staining? How was it fixed? Are you staining dewaxed and hydrated paraffin sections? Cryostat sections? Which haemalum & eosin method did you use? Does your lab have a book with instructions for H&E staining? If not, why not? Are the "faint stained spots on sections" blue, purple, pink or red? Where are these spots? John Kiernan Anatomy, UWO London, Canada. === ----- Original Message ----- From: Toxicology Date: Thursday, November 8, 2007 1:36 Subject: [Histonet] Artefacts To: histonet@lists.utsouthwestern.edu > Dear all, > I am facing a problem in Hematoxylin and Eosin staining. There are > several faint stained spots on sections and because of that its > difficuly to interpret the lesions. I am unable to sort out the > problem. Can any one help me? > > > > > -------------------------Email Disclaimer------------------------ > --------- > _______________________________________________ Histonet mailing list Histonet@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 Joseph's Health System, Inc. From Rcartun <@t> harthosp.org Thu Nov 8 11:36:02 2007 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Thu Nov 8 11:36:18 2007 Subject: [Histonet] B5 substitute In-Reply-To: References: Message-ID: <473302B20200007700009021@gwmail4.harthosp.org> Formalin (fix well and cut it thin). 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 >>> "Joanne Mauger" 11/08/07 11:50 AM >>> Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ 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 Rcartun <@t> harthosp.org Thu Nov 8 11:40:02 2007 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Thu Nov 8 11:40:25 2007 Subject: [Histonet] Clone for Prog. Receptor In-Reply-To: References: Message-ID: <473303A20200007700009027@gwmail4.harthosp.org> We have used clone PgR636 (from Dako) for several years now with excellent results. 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 >>> "Drew Sally A." 11/08/07 12:07 PM >>> We currently are using the 1A6 clone for our progesterone receptor(PR) IHC. We were wondering how other people felt about rabbit monoclonal PR antibodies. Does anyone have strong preferences or experiences with other PR clones-mouse or rabbit-that they'd like to share? Thank you...! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 _______________________________________________ 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 mtarango <@t> nvcancer.org Thu Nov 8 11:45:24 2007 From: mtarango <@t> nvcancer.org (Tarango, Mark) Date: Thu Nov 8 11:46:23 2007 Subject: [Histonet] problem in dual staining of CD4 and FOXP3 In-Reply-To: <813060655.505211194542002919.JavaMail.osg@osgjas04.cns.ufl.edu> Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F71@NVCIEXCH02.NVCI.org> Looking at T-regulatory cells, huh? Are you trying to stain both in the same color? I know Foxp3 is nuclear and CD4 is on the cell membrane, but two colors might be better. Your protocol looks incomplete. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of FU,DONGTAO Sent: Thursday, November 08, 2007 9:13 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3 Hello, Can anyone give me some suggestions on my case? I know a lot of people here have a lot of dual staining experiences. I did dual staining on 2 antibodies from different species in the past. They worked well. But using 2 antibodies from same species is my first time try and I met a big problem. Any suggestions I will be very appreciate. Ann On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" wrote: > Hi, all > > Thank you first for giving me some good suggestions on > thick-section question I posted last time. Now I met another > problem when I did CD4 and FOXP3 dual staining using murine fresh > frozen spleen. If I did single staining, both antibodies worked > very well. However if I did dual staining, I could only get good > result from the first antibody. The second one did not work at > all(I mean no specific staining). I used CD4 from BD(rat > anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. > I think there might be something wrong with my serum block(I used > normal rat serum block) before I added secondary antibody. Or any > other serum block I need to add to decrease the non-specific > binding which I have not done yet. Does anyone can give me some > suggestions according to my protocol below? How can I get > specific staining of the secondary (primary)antibody? Thank you, > > Below is the simple protocol I used for dual staining: > 1. 2% normal goat serum block 20 min > 2. 1* antibody CD4 1:750 in diluent O/N 4C > 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT > 4. Serum block: 5% normal rat serum 30 min > 5. 1* antibody FOXP3 1:100 in diluent 1h RT > 6. Secondary AF488 Donkey anti-rat in TBS 1h RT > > Use 1xTBS as wash buffer. Before staining, fix tissue in -20C > Aceton for 5 min, then airdry. > > > Ann Dongtao Fu MD Ph.D > Lab Manager > Molecular Pathology core > Dept. of Pathology > University of Florida > 1600 SW Archer Road > Gainesville, FL 32610 > Lab Phone: 352-273-7752 > FAX: 352-273-7753 > Rm: D11-50 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Ann Dongtao Fu MD, Ph.D Lab Manager Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== From mtarango <@t> nvcancer.org Thu Nov 8 11:47:54 2007 From: mtarango <@t> nvcancer.org (Tarango, Mark) Date: Thu Nov 8 11:48:13 2007 Subject: [Histonet] B5 substitute In-Reply-To: Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F72@NVCIEXCH02.NVCI.org> Why not just use 10% NBF? It's great if you allow enough time for fixation. Make sure your bone marrows are fixed well BEFORE decalcifying and you should have no problems. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joanne Mauger Sent: Thursday, November 08, 2007 8:51 AM To: gvdobbin@ihis.org; Histonet@lists.utsouthwestern.edu Subject: [Histonet] B5 substitute Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== From sjchtascp <@t> yahoo.com Thu Nov 8 11:58:22 2007 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Thu Nov 8 11:58:27 2007 Subject: [Histonet] HT position wanted Message-ID: <621094.56475.qm@web38209.mail.mud.yahoo.com> I'm looking for an HT position in the Madison, WI to Rockford, Ill area. Contract in Milwaukee, WI. Thanks, Steve __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From fudo <@t> ufl.edu Thu Nov 8 11:59:10 2007 From: fudo <@t> ufl.edu (FU,DONGTAO) Date: Thu Nov 8 11:59:20 2007 Subject: [Histonet] problem in dual staining of CD4 and FOXP3 Message-ID: <1211976397.304891194544750373.JavaMail.osg@osgjas03.cns.ufl.edu> Hi, Mark, Yes, I am looking at T-regulatory cells of mouse. For CD4, I used AF594(red color), for FOXP3 I used AF488(green color). The problem for me is for the second primary antibody(or maybe I should say for the secondary color) of dual staining, it never worked. I do not know which serum I should use to block the non-specific staining of the secondary antibody. If you think my protocol is imcomplete, could give me some suggestions? Many thanks, Ann On Thu Nov 08 12:45:24 EST 2007, "Tarango, Mark" wrote: > Looking at T-regulatory cells, huh? Are you trying to stain both > in the > same color? I know Foxp3 is nuclear and CD4 is on the cell > membrane, > but two colors might be better. Your protocol looks incomplete. > Mark Adam Tarango HT(ASCP) > Histology & IHC Supervisor > Nevada Cancer Institute > One Breakthrough Way > Las Vegas, NV 89135 > Direct Line (702) 822-5112 > Treo (702) 759-9229 > Fax (702) 939-7663 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > FU,DONGTAO > Sent: Thursday, November 08, 2007 9:13 AM > To: Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3 > > Hello, > > Can anyone give me some suggestions on my case? I know a lot of > people here have a lot of dual staining experiences. I did dual > staining on 2 antibodies from different species in the past. They > worked well. But using 2 antibodies from same species is my first > time try and I met a big problem. Any suggestions I will be very > appreciate. > > Ann > > > On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" > wrote: > >> Hi, all >> >> Thank you first for giving me some good suggestions on >> thick-section question I posted last time. Now I met another >> problem when I did CD4 and FOXP3 dual staining using murine >> fresh frozen spleen. If I did single staining, both antibodies >> worked very well. However if I did dual staining, I could only >> get good result from the first antibody. The second one did not >> work at all(I mean no specific staining). I used CD4 from BD(rat >> anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. >> I think there might be something wrong with my serum block(I >> used normal rat serum block) before I added secondary antibody. >> Or any other serum block I need to add to decrease the >> non-specific binding which I have not done yet. Does anyone can >> give me some suggestions according to my protocol below? How can >> I get specific staining of the secondary (primary)antibody? >> Thank you, >> >> Below is the simple protocol I used for dual staining: >> 1. 2% normal goat serum block 20 min >> 2. 1* antibody CD4 1:750 in diluent O/N 4C >> 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT >> 4. Serum block: 5% normal rat serum 30 min >> 5. 1* antibody FOXP3 1:100 in diluent 1h RT >> 6. Secondary AF488 Donkey anti-rat in TBS 1h RT >> >> Use 1xTBS as wash buffer. Before staining, fix tissue in -20C >> Aceton for 5 min, then airdry. >> >> >> Ann Dongtao Fu MD Ph.D >> Lab Manager >> Molecular Pathology core >> Dept. of Pathology >> University of Florida >> 1600 SW Archer Road >> Gainesville, FL 32610 >> Lab Phone: 352-273-7752 >> FAX: 352-273-7753 >> Rm: D11-50 >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > > > Ann Dongtao Fu MD, Ph.D > Lab Manager > Dept. of Pathology > Lab phone: 352-273-7752 > Lab FAX: 352-273-7755 > Lab address: D11-50 > PO Box: 100275 > 1600 SW Archer Road > University of Flodrida > Gainesville, FL 32610 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > "EMF " made the following annotations. > ------------------------------------------------------------------------------ > CONFIDENTIALITY NOTICE: This e-mail message, including any > attachments, is for the sole use of the intended recipient(s) and > may contain confidential, proprietary, and/or privileged > information protected by law. If you are not the intended > recipient, you may not use, copy, or distribute this e-mail > message or its attachments. If you believe you have received this > e-mail message in error, please contact the sender by reply > e-mail and destroy all copies of the original message > ============================================================================== > > > Ann Dongtao Fu MD, Ph.D Lab Manager Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 From rjbuesa <@t> yahoo.com Thu Nov 8 12:07:36 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 8 12:07:40 2007 Subject: [Histonet] B5 substitute In-Reply-To: <5AEC610C1CE02945BD63A395BA763EDE011B6F72@NVCIEXCH02.NVCI.org> Message-ID: <19687.42178.qm@web61218.mail.yahoo.com> Amen! Only make absolutely sure that the pH of your NBF is neutral (7). Ren? J. "Tarango, Mark" wrote: Why not just use 10% NBF? It's great if you allow enough time for fixation. Make sure your bone marrows are fixed well BEFORE decalcifying and you should have no problems. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joanne Mauger Sent: Thursday, November 08, 2007 8:51 AM To: gvdobbin@ihis.org; Histonet@lists.utsouthwestern.edu Subject: [Histonet] B5 substitute Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From arnie.jimenez <@t> vel-lab.com Thu Nov 8 12:14:42 2007 From: arnie.jimenez <@t> vel-lab.com (arnie.jimenez@vel-lab.com) Date: Thu Nov 8 12:16:56 2007 Subject: [Histonet] Re: Away from Office Message-ID: <20071108181442.24244.qmail@plesk5.hostgator.com> Thank you for contacting Vel-Lab Research. We will be out of the lab starting Nov. 7th and will return on Nov. 12th. We will check e-mail regularly but will be unable to respond. Please do not mail any packages to our lab until Nov. 12th. Regards, Arnie Jimenez Vel-Lab Research From b-frederick <@t> northwestern.edu Thu Nov 8 12:36:23 2007 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Thu Nov 8 12:36:40 2007 Subject: [Histonet] B5 substitute In-Reply-To: Message-ID: <000001c82236$46879c50$d00f7ca5@lurie.northwestern.edu> AZF works well. Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joanne Mauger Sent: Thursday, November 08, 2007 10:51 AM To: gvdobbin@ihis.org; Histonet@lists.utsouthwestern.edu Subject: [Histonet] B5 substitute Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From b-frederick <@t> northwestern.edu Thu Nov 8 12:38:12 2007 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Thu Nov 8 12:38:25 2007 Subject: [Histonet] Clone for Prog. Receptor In-Reply-To: Message-ID: <000101c82236$8754f750$d00f7ca5@lurie.northwestern.edu> Per a lecture at the NSh mouse monocloncal antibodies have better affinity than rabbit monoclonals. We use 1A6 but then, we are following a clinical protocol and cannot deviate. Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Drew Sally A. Sent: Thursday, November 08, 2007 11:07 AM To: Histonet Subject: [Histonet] Clone for Prog. Receptor We currently are using the 1A6 clone for our progesterone receptor(PR) IHC. We were wondering how other people felt about rabbit monoclonal PR antibodies. Does anyone have strong preferences or experiences with other PR clones-mouse or rabbit-that they'd like to share? Thank you...! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mtarango <@t> nvcancer.org Thu Nov 8 13:16:35 2007 From: mtarango <@t> nvcancer.org (Tarango, Mark) Date: Thu Nov 8 13:16:56 2007 Subject: [Histonet] problem in dual staining of CD4 and FOXP3 In-Reply-To: <1211976397.304891194544750373.JavaMail.osg@osgjas03.cns.ufl.edu> Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F73@NVCIEXCH02.NVCI.org> I didn't look closely enough at your protocol. I think you should omit step 4 of your protocol. Maybe when you trying blocking with rat serum the AF594 gets eaten up, since it is an anti-rat antibody. It might just go into the serum and wash away. ...Wait a sec, you said you were having problems with the second primary antibody.... I don't know what to say, try direct conjugates. Sorry I can't be of more help. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of FU,DONGTAO Sent: Thursday, November 08, 2007 9:59 AM To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] problem in dual staining of CD4 and FOXP3 Hi, Mark, Yes, I am looking at T-regulatory cells of mouse. For CD4, I used AF594(red color), for FOXP3 I used AF488(green color). The problem for me is for the second primary antibody(or maybe I should say for the secondary color) of dual staining, it never worked. I do not know which serum I should use to block the non-specific staining of the secondary antibody. If you think my protocol is imcomplete, could give me some suggestions? Many thanks, Ann On Thu Nov 08 12:45:24 EST 2007, "Tarango, Mark" wrote: > Looking at T-regulatory cells, huh? Are you trying to stain both > in the > same color? I know Foxp3 is nuclear and CD4 is on the cell > membrane, > but two colors might be better. Your protocol looks incomplete. > Mark Adam Tarango HT(ASCP) > Histology & IHC Supervisor > Nevada Cancer Institute > One Breakthrough Way > Las Vegas, NV 89135 > Direct Line (702) 822-5112 > Treo (702) 759-9229 > Fax (702) 939-7663 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > FU,DONGTAO > Sent: Thursday, November 08, 2007 9:13 AM > To: Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3 > > Hello, > > Can anyone give me some suggestions on my case? I know a lot of > people here have a lot of dual staining experiences. I did dual > staining on 2 antibodies from different species in the past. They > worked well. But using 2 antibodies from same species is my first > time try and I met a big problem. Any suggestions I will be very > appreciate. > > Ann > > > On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" > wrote: > >> Hi, all >> >> Thank you first for giving me some good suggestions on >> thick-section question I posted last time. Now I met another >> problem when I did CD4 and FOXP3 dual staining using murine >> fresh frozen spleen. If I did single staining, both antibodies >> worked very well. However if I did dual staining, I could only >> get good result from the first antibody. The second one did not >> work at all(I mean no specific staining). I used CD4 from BD(rat >> anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. >> I think there might be something wrong with my serum block(I >> used normal rat serum block) before I added secondary antibody. >> Or any other serum block I need to add to decrease the >> non-specific binding which I have not done yet. Does anyone can >> give me some suggestions according to my protocol below? How can >> I get specific staining of the secondary (primary)antibody? >> Thank you, >> >> Below is the simple protocol I used for dual staining: >> 1. 2% normal goat serum block 20 min >> 2. 1* antibody CD4 1:750 in diluent O/N 4C >> 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT >> 4. Serum block: 5% normal rat serum 30 min >> 5. 1* antibody FOXP3 1:100 in diluent 1h RT >> 6. Secondary AF488 Donkey anti-rat in TBS 1h RT >> >> Use 1xTBS as wash buffer. Before staining, fix tissue in -20C >> Aceton for 5 min, then airdry. >> >> >> Ann Dongtao Fu MD Ph.D >> Lab Manager >> Molecular Pathology core >> Dept. of Pathology >> University of Florida >> 1600 SW Archer Road >> Gainesville, FL 32610 >> Lab Phone: 352-273-7752 >> FAX: 352-273-7753 >> Rm: D11-50 >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > > > Ann Dongtao Fu MD, Ph.D > Lab Manager > Dept. of Pathology > Lab phone: 352-273-7752 > Lab FAX: 352-273-7755 > Lab address: D11-50 > PO Box: 100275 > 1600 SW Archer Road > University of Flodrida > Gainesville, FL 32610 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > "EMF " made the following annotations. > ------------------------------------------------------------------------ ------ > CONFIDENTIALITY NOTICE: This e-mail message, including any > attachments, is for the sole use of the intended recipient(s) and > may contain confidential, proprietary, and/or privileged > information protected by law. If you are not the intended > recipient, you may not use, copy, or distribute this e-mail > message or its attachments. If you believe you have received this > e-mail message in error, please contact the sender by reply > e-mail and destroy all copies of the original message > ======================================================================== ====== > > > Ann Dongtao Fu MD, Ph.D Lab Manager Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== From drbugge <@t> gmail.com Thu Nov 8 13:45:54 2007 From: drbugge <@t> gmail.com (Dawn Bugge) Date: Thu Nov 8 13:46:00 2007 Subject: [Histonet] Hematoxylin to dark on some slides Message-ID: <1c4db3750711081145h474f017g7aa568e8721cd676@mail.gmail.com> Recently we have been cutting some of our slides at night due to the high volume we have. What we are finding though is the slides that are dried over night are much darker then the slides we cut the same day. We have decreased the hematoxylin time on the slides done at night but haven't seen much difference. Any suggestions? -- Dawn R Bugge Seattle Histology From GDawson <@t> dynacaremilwaukee.com Thu Nov 8 13:47:40 2007 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Thu Nov 8 13:47:44 2007 Subject: FW: [Histonet] Re: Formaline-free fixative Message-ID: -----Original Message----- From: Dawson, Glen Sent: Thursday, November 08, 2007 1:46 PM To: 'Robert Richmond' Subject: RE: [Histonet] Re: Formaline-free fixative All, I know I've said this already but I think it merits repeating. Rather than looking into formalin alternatives, it would be best if ALL of us concentrated on using formalin wisely and looking for the best ways to deal with any problems that arise from its use. In today's climate of pushing standardized testing for world-wide, uniform results (which is impossible BTW), those who insist on using fixatives other than formalin are begging for a nightmare and doing a dis-service to their patients. Without a universal fixative, standardization of results from testing such as IHC is nothing but a pipe dream. Also, could you imagine trying to run an IHC reference lab and needing to have appropriate controls fixed in every conceivable fixative out there? Let's get realistic about this fixation issue because if the first step in histology (fixation) can't be solid, then all of the testing, staining, etc... that comes after is suspect. If your lab does all of its own testing and would never have to send a block out for a test that is not "in-house", I can see using whatever fixative you want and optimizing every procedure you have to that specific one, but we all know that there will always be an instance where that block could need to be sent out. My Opinion, Glen Dawson IHC Manager Milwaukee, WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Robert Richmond Sent: Thursday, November 08, 2007 10:08 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Formaline-free fixative Pierre Chaumat (where are you, Pierre?) asks: >>Has anyone tested alternative solution to formaline fixation ? Has anyone switched to a new tissue fixative ? - I would like to share some experience.<< About the only alternative "fixative" that actually is a fixative is glyoxal, available under numerous trade names. Its fixative properties are somewhat different from those of formaldehyde. In particular, immunostains may require adjustment, and comparison with formaldehyde fixed tissue from the same case. In the USA, the breast cancer immunostains are required by the federal govenrment to be done on formaldehyde fixed tissue. Do not use any trade-named fixative whose composition is secret. Sometimes fairly adequate information can be obtained from the Materials Safety Data Sheet (MSDS), in the USA. Does France require similar documentation of hazardous materials? If so, what is this document called, and what is a good Web site to look some of them up? (I can read French.) Bob Richmond Samurai Pathologist Knoxville, Tennessee _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Melissa.Gonzalez <@t> cellgenesys.com Thu Nov 8 13:47:49 2007 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Thu Nov 8 13:48:01 2007 Subject: [Histonet] RE: problem in dual staining of CD4 and FOXP3 Message-ID: <2884B897182A1D438C7BA24B9A8F94A213A580@hqsvr01mail.cgi.com> Hi Ann, I think the problem is that you are blocking with rat serum. You wouldn't block with rat serum before your first primary, and you shouldn't for your second primary since both are rat antibodies. There are several ways you could try to accomplish dual staining of 2 rat antibodies. I think the easiest, since you are doing fluorescent on frozens, is use BD's rat x mouse CD4 FITC (green cytoplasm). It works very well. Ebioscience has AF conjugated primaries, so you should be able to get rat x FOXP3 AF555 or AF594 (red nucleus). Then you could just cocktail them and do single step staining. Or you could try Probes (Invitrogen's) Zenon labeling kits, and directly conjugate your rat antibodies yourself. (I don't know if they have kits for rat primaries though) Thirdly, you could try following your protocol for the first primary, adding Biocare Medicals' Denaturing solution to seal the antigen site (since the first is cytoplasmic) then the second would have access to the nuclear site. 1. 5% normal goat serum block 20 min 2. 1* antibody CD4 1:750 in diluent O/N 4C 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT 4. Biocare Medicals' Denaturing solution (follow manuf instructions) 5. Serum block: 5% normal DONKEY serum 30 min 6. 1* antibody FOXP3 1:100 in diluent 1h RT 7. Secondary AF488 Donkey anti-rat in TBS 1h RT GL, Melissa Melissa A. Gonz?lez Edick R&D, Cell Genesys Inc. 500 Forbes Blvd South San Francisco, CA 94080 p(650) 266-3168 f (650) 266-3080 ---------------------------------------------------------------------------------------------------------- Message: 21 Date: Thu, 8 Nov 2007 12:59:10 -0500 (EST) From: "FU,DONGTAO" Subject: RE: [Histonet] problem in dual staining of CD4 and FOXP3 To: Histonet@lists.utsouthwestern.edu Message-ID: <1211976397.304891194544750373.JavaMail.osg@osgjas03.cns.ufl.edu> Content-Type: text/plain; format=flowed; charset=us-ascii Hi, Mark, Yes, I am looking at T-regulatory cells of mouse. For CD4, I used AF594(red color), for FOXP3 I used AF488(green color). The problem for me is for the second primary antibody(or maybe I should say for the secondary color) of dual staining, it never worked. I do not know which serum I should use to block the non-specific staining of the secondary antibody. If you think my protocol is imcomplete, could give me some suggestions? Many thanks, Ann On Thu Nov 08 12:45:24 EST 2007, "Tarango, Mark" wrote: > Looking at T-regulatory cells, huh? Are you trying to stain both > in the > same color? I know Foxp3 is nuclear and CD4 is on the cell > membrane, > but two colors might be better. Your protocol looks incomplete. > Mark Adam Tarango HT(ASCP) > Histology & IHC Supervisor > Nevada Cancer Institute > One Breakthrough Way > Las Vegas, NV 89135 > Direct Line (702) 822-5112 > Treo (702) 759-9229 > Fax (702) 939-7663 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > FU,DONGTAO > Sent: Thursday, November 08, 2007 9:13 AM > To: Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3 > > Hello, > > Can anyone give me some suggestions on my case? I know a lot of > people here have a lot of dual staining experiences. I did dual > staining on 2 antibodies from different species in the past. They > worked well. But using 2 antibodies from same species is my first > time try and I met a big problem. Any suggestions I will be very > appreciate. > > Ann > > > On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" > wrote: > >> Hi, all >> >> Thank you first for giving me some good suggestions on >> thick-section question I posted last time. Now I met another >> problem when I did CD4 and FOXP3 dual staining using murine >> fresh frozen spleen. If I did single staining, both antibodies >> worked very well. However if I did dual staining, I could only >> get good result from the first antibody. The second one did not >> work at all(I mean no specific staining). I used CD4 from BD(rat >> anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. >> I think there might be something wrong with my serum block(I >> used normal rat serum block) before I added secondary antibody. >> Or any other serum block I need to add to decrease the >> non-specific binding which I have not done yet. Does anyone can >> give me some suggestions according to my protocol below? How can >> I get specific staining of the secondary (primary)antibody? >> Thank you, >> >> Below is the simple protocol I used for dual staining: >> 1. 2% normal goat serum block 20 min >> 2. 1* antibody CD4 1:750 in diluent O/N 4C >> 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT >> 4. Serum block: 5% normal rat serum 30 min >> 5. 1* antibody FOXP3 1:100 in diluent 1h RT >> 6. Secondary AF488 Donkey anti-rat in TBS 1h RT From trinimaican2501 <@t> yahoo.com Thu Nov 8 14:00:50 2007 From: trinimaican2501 <@t> yahoo.com (I-sanna Gibbons) Date: Thu Nov 8 14:00:58 2007 Subject: [Histonet] frozen brain Message-ID: <254923.82101.qm@web50311.mail.re2.yahoo.com> Hi all, Does anyone have a protocol for freezing fixed (10% formalin) brain tissue? Will cell morphology be compromised to a large extent? Thanks I-sanna __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From michelle-griffin-reyes <@t> uiowa.edu Thu Nov 8 14:37:23 2007 From: michelle-griffin-reyes <@t> uiowa.edu (Griffin-Reyes, Michelle A) Date: Thu Nov 8 14:37:37 2007 Subject: [Histonet] RE: Mouse Histology Atlas In-Reply-To: <200711081823.lA8INVta015069@sun.its.uiowa.edu> Message-ID: I really like the book I am currently using. The title is "The Laboratory Mouse" edited by Hans Hedrich and Gillian Bullock. 2004. ISBN:0-12-336425-6. It goes over strains, stocks, morphologies, pros/cons to certain necropsy and sectioning methods. Hope this was of some help. Michelle A. Griffin-Reyes Comparative Pathology Laboratory University of Iowa Hospitals and Clinics -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Thursday, November 08, 2007 12:24 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 48, Issue 11 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. mouse histology atlas (Helen E Johnson) 2. Re: Away from Office (arnie.jimenez@vel-lab.com) 3. Short processing schedule-suggestions (Greg Dobbin) 4. Re: Formaline-free fixative (Robert Richmond) 5. B5 substitute (Joanne Mauger) 6. RE: Artefacts (Rene J Buesa) 7. Re: Short processing schedule-suggestions (Rene J Buesa) 8. Clone for Prog. Receptor (Drew Sally A.) 9. Re: Artefacts (John Kiernan) 10. RE: Artefacts (Marshall Terry Dr, Consultant Histopathologist) 11. Re: problem in dual staining of CD4 and FOXP3 (FU,DONGTAO) 12. RE: Artefacts (Weems, Joyce) 13. RE: B5 substitute (Weems, Joyce) 14. Re: B5 substitute (Matt Bancroft) 15. RE: Artefacts (Marshall Terry Dr, Consultant Histopathologist) 16. Re: B5 substitute (Richard Cartun) 17. Re: Clone for Prog. Receptor (Richard Cartun) 18. RE: problem in dual staining of CD4 and FOXP3 (Tarango, Mark) 19. RE: B5 substitute (Tarango, Mark) 20. HT position wanted (Steven Coakley) 21. RE: problem in dual staining of CD4 and FOXP3 (FU,DONGTAO) ---------------------------------------------------------------------- Message: 1 Date: Thu, 8 Nov 2007 09:22:44 -0500 From: Helen E Johnson Subject: [Histonet] mouse histology atlas To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Histonetters, Does anyone know of a good mouse histology atlas? Helen Johnson (hej01@health.state.ny.us) IMPORTANT NOTICE: This e-mail and any attachments may contain confidential or sensitive information which is, or may be, legally privileged or otherwise protected by law from further disclosure. It is intended only for the addressee. If you received this in error or from someone who was not authorized to send it to you, please do not distribute, copy or use it or any attachments. Please notify the sender immediately by reply e-mail and delete this from your system. Thank you for your cooperation. ------------------------------ Message: 2 Date: 8 Nov 2007 08:23:20 -0600 From: arnie.jimenez@vel-lab.com Subject: [Histonet] Re: Away from Office To: histonet@lists.utsouthwestern.edu Message-ID: <20071108142320.15276.qmail@plesk5.hostgator.com> Content-Type: text/plain; charset="UTF-8" Thank you for contacting Vel-Lab Research. We will be out of the lab starting Nov. 7th and will return on Nov. 12th. We will check e-mail regularly but will be unable to respond. Please do not mail any packages to our lab until Nov. 12th. Regards, Arnie Jimenez Vel-Lab Research ------------------------------ Message: 3 Date: Thu, 08 Nov 2007 10:59:21 -0400 From: "Greg Dobbin" Subject: [Histonet] Short processing schedule-suggestions To: Message-ID: Content-Type: text/plain; charset=US-ASCII Hello All, I would like to get your suggestions for a processing schedule for small biopsies (cores, GI's, other endoscopics). I want to process these little guys separate from everything else to hopefully improve section quality and perhaps staining quality. I don't want the absolute shortest possible schedule. We have a newer VIP5 for our main processor (doing everything on one schedule currently) and a much older VIP sitting idle as a backup that I plan to start using for the biopsies. We process overnight so I have more than enough time. I will just delay the biopsy processing so that both processors are finished at essentially the same time. What I need is a shorter schedule for the biopsies so that the detrimental affects of xylene and alcohol are minimized (and yes, we still use xylene! And in case it matters schedule-wise, we use 10% NBFS as well). Thanks in advance. Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. ------------------------------ Message: 4 Date: Thu, 8 Nov 2007 10:08:27 -0600 From: "Robert Richmond" Subject: [Histonet] Re: Formaline-free fixative To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Pierre Chaumat (where are you, Pierre?) asks: >>Has anyone tested alternative solution to formaline fixation ? Has anyone switched to a new tissue fixative ? - I would like to share some experience.<< About the only alternative "fixative" that actually is a fixative is glyoxal, available under numerous trade names. Its fixative properties are somewhat different from those of formaldehyde. In particular, immunostains may require adjustment, and comparison with formaldehyde fixed tissue from the same case. In the USA, the breast cancer immunostains are required by the federal govenrment to be done on formaldehyde fixed tissue. Do not use any trade-named fixative whose composition is secret. Sometimes fairly adequate information can be obtained from the Materials Safety Data Sheet (MSDS), in the USA. Does France require similar documentation of hazardous materials? If so, what is this document called, and what is a good Web site to look some of them up? (I can read French.) Bob Richmond Samurai Pathologist Knoxville, Tennessee ------------------------------ Message: 5 Date: Thu, 08 Nov 2007 11:50:43 -0500 From: "Joanne Mauger" Subject: [Histonet] B5 substitute To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger ------------------------------ Message: 6 Date: Thu, 8 Nov 2007 08:59:40 -0800 (PST) From: Rene J Buesa Subject: RE: [Histonet] Artefacts To: Kemlo Rogerson , Toxicology , histonet@lists.utsouthwestern.edu Message-ID: <222219.92551.qm@web61219.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 and most likely the first! Ren? J. Kemlo Rogerson wrote: Ah the spotty H&E problem again. 1) Inadequate dewaxing. 2) Inadequate fixation. 3) Inadequate processing. Not necessarily in that order. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation _______________________________________________ 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 ------------------------------ Message: 7 Date: Thu, 8 Nov 2007 09:03:38 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Short processing schedule-suggestions To: Greg Dobbin , Histonet@lists.utsouthwestern.edu Message-ID: <462982.42437.qm@web61214.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 >From my experience in processing, the most important thing is to maintain dehydration time to clearing time to infiltration time in constant proportions once you find a good protocol. If you "long" protocol works well for you, reduce the time in each step in a way that at the end will be completed in the time you need, BUT preserving the time proportions you have now. Ren? J. Greg Dobbin wrote: Hello All, I would like to get your suggestions for a processing schedule for small biopsies (cores, GI's, other endoscopics). I want to process these little guys separate from everything else to hopefully improve section quality and perhaps staining quality. I don't want the absolute shortest possible schedule. We have a newer VIP5 for our main processor (doing everything on one schedule currently) and a much older VIP sitting idle as a backup that I plan to start using for the biopsies. We process overnight so I have more than enough time. I will just delay the biopsy processing so that both processors are finished at essentially the same time. What I need is a shorter schedule for the biopsies so that the detrimental affects of xylene and alcohol are minimized (and yes, we still use xylene! And in case it matters schedule-wise, we use 10% NBFS as well). Thanks in advance. Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. _______________________________________________ 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 ------------------------------ Message: 8 Date: Thu, 8 Nov 2007 11:07:25 -0600 From: "Drew Sally A." Subject: [Histonet] Clone for Prog. Receptor To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" We currently are using the 1A6 clone for our progesterone receptor(PR) IHC. We were wondering how other people felt about rabbit monoclonal PR antibodies. Does anyone have strong preferences or experiences with other PR clones-mouse or rabbit-that they'd like to share? Thank you...! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 ------------------------------ Message: 9 Date: Thu, 08 Nov 2007 12:10:47 -0500 From: John Kiernan Subject: Re: [Histonet] Artefacts To: Toxicology Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Dear Toxicology You need to explain your problem more clearly. Please include histonet@lists.utsouthwestern.edu in your replies. We all want to share our knowledge. What tissue are you staining? How was it fixed? Are you staining dewaxed and hydrated paraffin sections? Cryostat sections? Which haemalum & eosin method did you use? Does your lab have a book with instructions for H&E staining? If not, why not? Are the "faint stained spots on sections" blue, purple, pink or red? Where are these spots? John Kiernan Anatomy, UWO London, Canada. === ----- Original Message ----- From: Toxicology Date: Thursday, November 8, 2007 1:36 Subject: [Histonet] Artefacts To: histonet@lists.utsouthwestern.edu > Dear all, > I am facing a problem in Hematoxylin and Eosin staining. There > are several > faint stained spots on sections and because of that its > difficuly to > interpret the lesions. I am unable to sort out the > problem. Can any one > help me? > > > > > -------------------------Email Disclaimer------------------------ > --------- > ------------------------------ Message: 10 Date: Thu, 8 Nov 2007 17:13:21 -0000 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Artefacts To: "John Kiernan" , "Toxicology" Cc: histonet@lists.utsouthwestern.edu Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE4F@TRFT-EX01.xRothGen.nhs.uk> Content-Type: text/plain; charset="us-ascii" Some of these questions should be answered by the photos, but I cannot find them. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: 08 November 2007 17:11 To: Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Artefacts Dear Toxicology You need to explain your problem more clearly. Please include histonet@lists.utsouthwestern.edu in your replies. We all want to share our knowledge. What tissue are you staining? How was it fixed? Are you staining dewaxed and hydrated paraffin sections? Cryostat sections? Which haemalum & eosin method did you use? Does your lab have a book with instructions for H&E staining? If not, why not? Are the "faint stained spots on sections" blue, purple, pink or red? Where are these spots? John Kiernan Anatomy, UWO London, Canada. === ----- Original Message ----- From: Toxicology Date: Thursday, November 8, 2007 1:36 Subject: [Histonet] Artefacts To: histonet@lists.utsouthwestern.edu > Dear all, > I am facing a problem in Hematoxylin and Eosin staining. There are > several faint stained spots on sections and because of that its > difficuly to interpret the lesions. I am unable to sort out the > problem. Can any one help me? > > > > > -------------------------Email Disclaimer------------------------ > --------- > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Thu, 8 Nov 2007 12:13:22 -0500 (EST) From: "FU,DONGTAO" Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3 To: Histonet@lists.utsouthwestern.edu Message-ID: <813060655.505211194542002919.JavaMail.osg@osgjas04.cns.ufl.edu> Content-Type: text/plain; format=flowed; charset=us-ascii Hello, Can anyone give me some suggestions on my case? I know a lot of people here have a lot of dual staining experiences. I did dual staining on 2 antibodies from different species in the past. They worked well. But using 2 antibodies from same species is my first time try and I met a big problem. Any suggestions I will be very appreciate. Ann On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" wrote: > Hi, all > > Thank you first for giving me some good suggestions on > thick-section question I posted last time. Now I met another > problem when I did CD4 and FOXP3 dual staining using murine fresh > frozen spleen. If I did single staining, both antibodies worked > very well. However if I did dual staining, I could only get good > result from the first antibody. The second one did not work at > all(I mean no specific staining). I used CD4 from BD(rat > anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. > I think there might be something wrong with my serum block(I used > normal rat serum block) before I added secondary antibody. Or any > other serum block I need to add to decrease the non-specific > binding which I have not done yet. Does anyone can give me some > suggestions according to my protocol below? How can I get > specific staining of the secondary (primary)antibody? Thank you, > > Below is the simple protocol I used for dual staining: > 1. 2% normal goat serum block 20 min > 2. 1* antibody CD4 1:750 in diluent O/N 4C > 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT > 4. Serum block: 5% normal rat serum 30 min > 5. 1* antibody FOXP3 1:100 in diluent 1h RT > 6. Secondary AF488 Donkey anti-rat in TBS 1h RT > > Use 1xTBS as wash buffer. Before staining, fix tissue in -20C > Aceton for 5 min, then airdry. > > > Ann Dongtao Fu MD Ph.D > Lab Manager > Molecular Pathology core > Dept. of Pathology > University of Florida > 1600 SW Archer Road > Gainesville, FL 32610 > Lab Phone: 352-273-7752 > FAX: 352-273-7753 > Rm: D11-50 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Ann Dongtao Fu MD, Ph.D Lab Manager Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 ------------------------------ Message: 12 Date: Thu, 8 Nov 2007 12:16:04 -0500 From: "Weems, Joyce" Subject: RE: [Histonet] Artefacts To: "Marshall Terry Dr, Consultant Histopathologist" , "John Kiernan" , "Toxicology" Cc: histonet@lists.utsouthwestern.edu Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD32048F41AF@sjhaexc02.sjha.org> Content-Type: text/plain; charset="utf-8" The web site says give it 24 hrs for the photo to post... Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, November 08, 2007 12:13 PM To: John Kiernan; Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Artefacts Some of these questions should be answered by the photos, but I cannot find them. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: 08 November 2007 17:11 To: Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Artefacts Dear Toxicology You need to explain your problem more clearly. Please include histonet@lists.utsouthwestern.edu in your replies. We all want to share our knowledge. What tissue are you staining? How was it fixed? Are you staining dewaxed and hydrated paraffin sections? Cryostat sections? Which haemalum & eosin method did you use? Does your lab have a book with instructions for H&E staining? If not, why not? Are the "faint stained spots on sections" blue, purple, pink or red? Where are these spots? John Kiernan Anatomy, UWO London, Canada. === ----- Original Message ----- From: Toxicology Date: Thursday, November 8, 2007 1:36 Subject: [Histonet] Artefacts To: histonet@lists.utsouthwestern.edu > Dear all, > I am facing a problem in Hematoxylin and Eosin staining. There are > several faint stained spots on sections and because of that its > difficuly to interpret the lesions. I am unable to sort out the > problem. Can any one help me? > > > > > -------------------------Email Disclaimer------------------------ > --------- > _______________________________________________ Histonet mailing list Histonet@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 Joseph's Health System, Inc. ------------------------------ Message: 13 Date: Thu, 8 Nov 2007 12:20:12 -0500 From: "Weems, Joyce" Subject: RE: [Histonet] B5 substitute To: Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD32048F41B0@sjhaexc02.sjha.org> Content-Type: text/plain; charset="utf-8" We use B-Plus from BBC. http://www.bbcus.com/website_search.html It takes a long time to get used to, especially when they have to compare to previous mercury fixed cases, but it can be done! Good luck, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne Mauger Sent: Thursday, November 08, 2007 11:51 AM To: gvdobbin@ihis.org; Histonet@lists.utsouthwestern.edu Subject: [Histonet] B5 substitute Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ 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 Joseph's Health System, Inc. ------------------------------ Message: 14 Date: Thu, 8 Nov 2007 09:24:17 -0800 (PST) From: Matt Bancroft Subject: Re: [Histonet] B5 substitute To: Joanne Mauger , gvdobbin@ihis.org, Histonet@lists.utsouthwestern.edu Message-ID: <704342.79971.qm@web63407.mail.re1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 B-Plus Joanne Mauger wrote: Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ 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 ------------------------------ Message: 15 Date: Thu, 8 Nov 2007 17:27:01 -0000 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Artefacts To: "Weems, Joyce" , "John Kiernan" , "Toxicology" Cc: histonet@lists.utsouthwestern.edu Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE50@TRFT-EX01.xRothGen.nhs.uk> Content-Type: text/plain; charset="us-ascii" Yes, I know. Posters can read that too and wait until the photos appear before they post the question. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: 08 November 2007 17:16 To: Marshall Terry Dr, Consultant Histopathologist; John Kiernan; Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Artefacts The web site says give it 24 hrs for the photo to post... Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, November 08, 2007 12:13 PM To: John Kiernan; Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Artefacts Some of these questions should be answered by the photos, but I cannot find them. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: 08 November 2007 17:11 To: Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Artefacts Dear Toxicology You need to explain your problem more clearly. Please include histonet@lists.utsouthwestern.edu in your replies. We all want to share our knowledge. What tissue are you staining? How was it fixed? Are you staining dewaxed and hydrated paraffin sections? Cryostat sections? Which haemalum & eosin method did you use? Does your lab have a book with instructions for H&E staining? If not, why not? Are the "faint stained spots on sections" blue, purple, pink or red? Where are these spots? John Kiernan Anatomy, UWO London, Canada. === ----- Original Message ----- From: Toxicology Date: Thursday, November 8, 2007 1:36 Subject: [Histonet] Artefacts To: histonet@lists.utsouthwestern.edu > Dear all, > I am facing a problem in Hematoxylin and Eosin staining. There are > several faint stained spots on sections and because of that its > difficuly to interpret the lesions. I am unable to sort out the > problem. Can any one help me? > > > > > -------------------------Email Disclaimer------------------------ > --------- > _______________________________________________ Histonet mailing list Histonet@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 Joseph's Health System, Inc. ------------------------------ Message: 16 Date: Thu, 08 Nov 2007 12:36:02 -0500 From: "Richard Cartun" Subject: Re: [Histonet] B5 substitute To: "Joanne Mauger" ,, Message-ID: <473302B20200007700009021@gwmail4.harthosp.org> Content-Type: text/plain; charset=US-ASCII Formalin (fix well and cut it thin). 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 >>> "Joanne Mauger" 11/08/07 11:50 AM >>> Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ 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: 17 Date: Thu, 08 Nov 2007 12:40:02 -0500 From: "Richard Cartun" Subject: Re: [Histonet] Clone for Prog. Receptor To: "Histonet" , "Drew Sally A." Message-ID: <473303A20200007700009027@gwmail4.harthosp.org> Content-Type: text/plain; charset=US-ASCII We have used clone PgR636 (from Dako) for several years now with excellent results. 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 >>> "Drew Sally A." 11/08/07 12:07 PM >>> We currently are using the 1A6 clone for our progesterone receptor(PR) IHC. We were wondering how other people felt about rabbit monoclonal PR antibodies. Does anyone have strong preferences or experiences with other PR clones-mouse or rabbit-that they'd like to share? Thank you...! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 _______________________________________________ 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: 18 Date: Thu, 8 Nov 2007 09:45:24 -0800 From: "Tarango, Mark" Subject: RE: [Histonet] problem in dual staining of CD4 and FOXP3 To: "FU,DONGTAO" , Histonet@lists.utsouthwestern.edu Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F71@NVCIEXCH02.NVCI.org> Content-Type: text/plain; charset=us-ascii Looking at T-regulatory cells, huh? Are you trying to stain both in the same color? I know Foxp3 is nuclear and CD4 is on the cell membrane, but two colors might be better. Your protocol looks incomplete. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of FU,DONGTAO Sent: Thursday, November 08, 2007 9:13 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3 Hello, Can anyone give me some suggestions on my case? I know a lot of people here have a lot of dual staining experiences. I did dual staining on 2 antibodies from different species in the past. They worked well. But using 2 antibodies from same species is my first time try and I met a big problem. Any suggestions I will be very appreciate. Ann On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" wrote: > Hi, all > > Thank you first for giving me some good suggestions on > thick-section question I posted last time. Now I met another > problem when I did CD4 and FOXP3 dual staining using murine fresh > frozen spleen. If I did single staining, both antibodies worked > very well. However if I did dual staining, I could only get good > result from the first antibody. The second one did not work at > all(I mean no specific staining). I used CD4 from BD(rat > anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. > I think there might be something wrong with my serum block(I used > normal rat serum block) before I added secondary antibody. Or any > other serum block I need to add to decrease the non-specific > binding which I have not done yet. Does anyone can give me some > suggestions according to my protocol below? How can I get > specific staining of the secondary (primary)antibody? Thank you, > > Below is the simple protocol I used for dual staining: > 1. 2% normal goat serum block 20 min > 2. 1* antibody CD4 1:750 in diluent O/N 4C > 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT > 4. Serum block: 5% normal rat serum 30 min > 5. 1* antibody FOXP3 1:100 in diluent 1h RT > 6. Secondary AF488 Donkey anti-rat in TBS 1h RT > > Use 1xTBS as wash buffer. Before staining, fix tissue in -20C > Aceton for 5 min, then airdry. > > > Ann Dongtao Fu MD Ph.D > Lab Manager > Molecular Pathology core > Dept. of Pathology > University of Florida > 1600 SW Archer Road > Gainesville, FL 32610 > Lab Phone: 352-273-7752 > FAX: 352-273-7753 > Rm: D11-50 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Ann Dongtao Fu MD, Ph.D Lab Manager Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== ------------------------------ Message: 19 Date: Thu, 8 Nov 2007 09:47:54 -0800 From: "Tarango, Mark" Subject: RE: [Histonet] B5 substitute To: "Joanne Mauger" , gvdobbin@ihis.org, Histonet@lists.utsouthwestern.edu Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F72@NVCIEXCH02.NVCI.org> Content-Type: text/plain; charset=us-ascii Why not just use 10% NBF? It's great if you allow enough time for fixation. Make sure your bone marrows are fixed well BEFORE decalcifying and you should have no problems. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joanne Mauger Sent: Thursday, November 08, 2007 8:51 AM To: gvdobbin@ihis.org; Histonet@lists.utsouthwestern.edu Subject: [Histonet] B5 substitute Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== ------------------------------ Message: 20 Date: Thu, 8 Nov 2007 09:58:22 -0800 (PST) From: Steven Coakley Subject: [Histonet] HT position wanted To: Histonet@lists.utsouthwestern.edu Message-ID: <621094.56475.qm@web38209.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I'm looking for an HT position in the Madison, WI to Rockford, Ill area. Contract in Milwaukee, WI. Thanks, Steve __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 21 Date: Thu, 8 Nov 2007 12:59:10 -0500 (EST) From: "FU,DONGTAO" Subject: RE: [Histonet] problem in dual staining of CD4 and FOXP3 To: Histonet@lists.utsouthwestern.edu Message-ID: <1211976397.304891194544750373.JavaMail.osg@osgjas03.cns.ufl.edu> Content-Type: text/plain; format=flowed; charset=us-ascii Hi, Mark, Yes, I am looking at T-regulatory cells of mouse. For CD4, I used AF594(red color), for FOXP3 I used AF488(green color). The problem for me is for the second primary antibody(or maybe I should say for the secondary color) of dual staining, it never worked. I do not know which serum I should use to block the non-specific staining of the secondary antibody. If you think my protocol is imcomplete, could give me some suggestions? Many thanks, Ann On Thu Nov 08 12:45:24 EST 2007, "Tarango, Mark" wrote: > Looking at T-regulatory cells, huh? Are you trying to stain both > in the > same color? I know Foxp3 is nuclear and CD4 is on the cell > membrane, > but two colors might be better. Your protocol looks incomplete. > Mark Adam Tarango HT(ASCP) > Histology & IHC Supervisor > Nevada Cancer Institute > One Breakthrough Way > Las Vegas, NV 89135 > Direct Line (702) 822-5112 > Treo (702) 759-9229 > Fax (702) 939-7663 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > FU,DONGTAO > Sent: Thursday, November 08, 2007 9:13 AM > To: Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3 > > Hello, > > Can anyone give me some suggestions on my case? I know a lot of > people here have a lot of dual staining experiences. I did dual > staining on 2 antibodies from different species in the past. They > worked well. But using 2 antibodies from same species is my first > time try and I met a big problem. Any suggestions I will be very > appreciate. > > Ann > > > On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" > wrote: > >> Hi, all >> >> Thank you first for giving me some good suggestions on >> thick-section question I posted last time. Now I met another >> problem when I did CD4 and FOXP3 dual staining using murine >> fresh frozen spleen. If I did single staining, both antibodies >> worked very well. However if I did dual staining, I could only >> get good result from the first antibody. The second one did not >> work at all(I mean no specific staining). I used CD4 from BD(rat >> anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. >> I think there might be something wrong with my serum block(I >> used normal rat serum block) before I added secondary antibody. >> Or any other serum block I need to add to decrease the >> non-specific binding which I have not done yet. Does anyone can >> give me some suggestions according to my protocol below? How can >> I get specific staining of the secondary (primary)antibody? >> Thank you, >> >> Below is the simple protocol I used for dual staining: >> 1. 2% normal goat serum block 20 min >> 2. 1* antibody CD4 1:750 in diluent O/N 4C >> 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT >> 4. Serum block: 5% normal rat serum 30 min >> 5. 1* antibody FOXP3 1:100 in diluent 1h RT >> 6. Secondary AF488 Donkey anti-rat in TBS 1h RT >> >> Use 1xTBS as wash buffer. Before staining, fix tissue in -20C >> Aceton for 5 min, then airdry. >> >> >> Ann Dongtao Fu MD Ph.D >> Lab Manager >> Molecular Pathology core >> Dept. of Pathology >> University of Florida >> 1600 SW Archer Road >> Gainesville, FL 32610 >> Lab Phone: 352-273-7752 >> FAX: 352-273-7753 >> Rm: D11-50 >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > > > Ann Dongtao Fu MD, Ph.D > Lab Manager > Dept. of Pathology > Lab phone: 352-273-7752 > Lab FAX: 352-273-7755 > Lab address: D11-50 > PO Box: 100275 > 1600 SW Archer Road > University of Flodrida > Gainesville, FL 32610 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > "EMF " made the following annotations. > ------------------------------------------------------------------------------ > CONFIDENTIALITY NOTICE: This e-mail message, including any > attachments, is for the sole use of the intended recipient(s) and > may contain confidential, proprietary, and/or privileged > information protected by law. If you are not the intended > recipient, you may not use, copy, or distribute this e-mail > message or its attachments. If you believe you have received this > e-mail message in error, please contact the sender by reply > e-mail and destroy all copies of the original message > ============================================================================== > > > Ann Dongtao Fu MD, Ph.D Lab Manager Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 11 **************************************** From kmilne <@t> bccancer.bc.ca Thu Nov 8 15:22:12 2007 From: kmilne <@t> bccancer.bc.ca (Milne, Katy) Date: Thu Nov 8 15:22:20 2007 Subject: [Histonet] Dendritic cells in mouse FFPE tissue In-Reply-To: <6e37ab$24kaug@ironport1in.cw.bc.ca> Message-ID: <07979E76B0869D4E8C9FE4AA9FC0657804668BC5@srvex03.phsabc.ehcnet.ca> Ok, I know I'm on another one of my wild goose chases again but is anyone successfully working with an antibody that detects dendritic cells in mouse FFPE tissue (no, frozen is not an option at this point)? It needs to be something specific for dendritic cells, i.e. no cross-reactivity with macrophages etc. Thanks, Katy Milne Deeley Research Centre From pierre.chaumat <@t> alphelys.com Thu Nov 8 16:16:08 2007 From: pierre.chaumat <@t> alphelys.com (Pierre CHAUMAT) Date: Thu Nov 8 16:16:31 2007 Subject: [Histonet] Formaline free fixative In-Reply-To: <200711081807.lA8I7GT1028397@smtp39.msg.oleane.net> Message-ID: <0ML2xA-1IqFfU2ECJ-0004wN@mrelayeu.kundenserver.de> Dear Bob, I know indeed quite a number of glyoxal based fixatives. Many labs have tested them over here. Products were FineFix, GlyoFix, amongst others. However, most of time morphology results were OK but IHC was bad. It looks like glyoxal tends to fix too much. As for MSDS, yes, they are required as well in France and I have noticed that allowed ppm in lab air is even less that with formalin ! Do you have yourself any experience with glyoxal based fixative ? Best regards Pierre -----Message d'origine----- De : histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] De la part de histonet-request@lists.utsouthwestern.edu Envoy? : jeudi 8 novembre 2007 19:07 ? : histonet@lists.utsouthwestern.edu Objet : Histonet Digest, Vol 48, Issue 11 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. mouse histology atlas (Helen E Johnson) 2. Re: Away from Office (arnie.jimenez@vel-lab.com) 3. Short processing schedule-suggestions (Greg Dobbin) 4. Re: Formaline-free fixative (Robert Richmond) 5. B5 substitute (Joanne Mauger) 6. RE: Artefacts (Rene J Buesa) 7. Re: Short processing schedule-suggestions (Rene J Buesa) 8. Clone for Prog. Receptor (Drew Sally A.) 9. Re: Artefacts (John Kiernan) 10. RE: Artefacts (Marshall Terry Dr, Consultant Histopathologist) 11. Re: problem in dual staining of CD4 and FOXP3 (FU,DONGTAO) 12. RE: Artefacts (Weems, Joyce) 13. RE: B5 substitute (Weems, Joyce) 14. Re: B5 substitute (Matt Bancroft) 15. RE: Artefacts (Marshall Terry Dr, Consultant Histopathologist) 16. Re: B5 substitute (Richard Cartun) 17. Re: Clone for Prog. Receptor (Richard Cartun) 18. RE: problem in dual staining of CD4 and FOXP3 (Tarango, Mark) 19. RE: B5 substitute (Tarango, Mark) 20. HT position wanted (Steven Coakley) 21. RE: problem in dual staining of CD4 and FOXP3 (FU,DONGTAO) ---------------------------------------------------------------------- Message: 1 Date: Thu, 8 Nov 2007 09:22:44 -0500 From: Helen E Johnson Subject: [Histonet] mouse histology atlas To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Histonetters, Does anyone know of a good mouse histology atlas? Helen Johnson (hej01@health.state.ny.us) IMPORTANT NOTICE: This e-mail and any attachments may contain confidential or sensitive information which is, or may be, legally privileged or otherwise protected by law from further disclosure. It is intended only for the addressee. If you received this in error or from someone who was not authorized to send it to you, please do not distribute, copy or use it or any attachments. Please notify the sender immediately by reply e-mail and delete this from your system. Thank you for your cooperation. ------------------------------ Message: 2 Date: 8 Nov 2007 08:23:20 -0600 From: arnie.jimenez@vel-lab.com Subject: [Histonet] Re: Away from Office To: histonet@lists.utsouthwestern.edu Message-ID: <20071108142320.15276.qmail@plesk5.hostgator.com> Content-Type: text/plain; charset="UTF-8" Thank you for contacting Vel-Lab Research. We will be out of the lab starting Nov. 7th and will return on Nov. 12th. We will check e-mail regularly but will be unable to respond. Please do not mail any packages to our lab until Nov. 12th. Regards, Arnie Jimenez Vel-Lab Research ------------------------------ Message: 3 Date: Thu, 08 Nov 2007 10:59:21 -0400 From: "Greg Dobbin" Subject: [Histonet] Short processing schedule-suggestions To: Message-ID: Content-Type: text/plain; charset=US-ASCII Hello All, I would like to get your suggestions for a processing schedule for small biopsies (cores, GI's, other endoscopics). I want to process these little guys separate from everything else to hopefully improve section quality and perhaps staining quality. I don't want the absolute shortest possible schedule. We have a newer VIP5 for our main processor (doing everything on one schedule currently) and a much older VIP sitting idle as a backup that I plan to start using for the biopsies. We process overnight so I have more than enough time. I will just delay the biopsy processing so that both processors are finished at essentially the same time. What I need is a shorter schedule for the biopsies so that the detrimental affects of xylene and alcohol are minimized (and yes, we still use xylene! And in case it matters schedule-wise, we use 10% NBFS as well). Thanks in advance. Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. ------------------------------ Message: 4 Date: Thu, 8 Nov 2007 10:08:27 -0600 From: "Robert Richmond" Subject: [Histonet] Re: Formaline-free fixative To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Pierre Chaumat (where are you, Pierre?) asks: >>Has anyone tested alternative solution to formaline fixation ? Has >>anyone switched to a new tissue fixative ? - I would like to share some experience.<< About the only alternative "fixative" that actually is a fixative is glyoxal, available under numerous trade names. Its fixative properties are somewhat different from those of formaldehyde. In particular, immunostains may require adjustment, and comparison with formaldehyde fixed tissue from the same case. In the USA, the breast cancer immunostains are required by the federal govenrment to be done on formaldehyde fixed tissue. Do not use any trade-named fixative whose composition is secret. Sometimes fairly adequate information can be obtained from the Materials Safety Data Sheet (MSDS), in the USA. Does France require similar documentation of hazardous materials? If so, what is this document called, and what is a good Web site to look some of them up? (I can read French.) Bob Richmond Samurai Pathologist Knoxville, Tennessee ------------------------------ Message: 5 Date: Thu, 08 Nov 2007 11:50:43 -0500 From: "Joanne Mauger" Subject: [Histonet] B5 substitute To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger ------------------------------ Message: 6 Date: Thu, 8 Nov 2007 08:59:40 -0800 (PST) From: Rene J Buesa Subject: RE: [Histonet] Artefacts To: Kemlo Rogerson , Toxicology , histonet@lists.utsouthwestern.edu Message-ID: <222219.92551.qm@web61219.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 and most likely the first! Reni J. Kemlo Rogerson wrote: Ah the spotty H&E problem again. 1) Inadequate dewaxing. 2) Inadequate fixation. 3) Inadequate processing. Not necessarily in that order. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation _______________________________________________ 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 ------------------------------ Message: 7 Date: Thu, 8 Nov 2007 09:03:38 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Short processing schedule-suggestions To: Greg Dobbin , Histonet@lists.utsouthwestern.edu Message-ID: <462982.42437.qm@web61214.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 >From my experience in processing, the most important thing is to maintain dehydration time to clearing time to infiltration time in constant proportions once you find a good protocol. If you "long" protocol works well for you, reduce the time in each step in a way that at the end will be completed in the time you need, BUT preserving the time proportions you have now. Reni J. Greg Dobbin wrote: Hello All, I would like to get your suggestions for a processing schedule for small biopsies (cores, GI's, other endoscopics). I want to process these little guys separate from everything else to hopefully improve section quality and perhaps staining quality. I don't want the absolute shortest possible schedule. We have a newer VIP5 for our main processor (doing everything on one schedule currently) and a much older VIP sitting idle as a backup that I plan to start using for the biopsies. We process overnight so I have more than enough time. I will just delay the biopsy processing so that both processors are finished at essentially the same time. What I need is a shorter schedule for the biopsies so that the detrimental affects of xylene and alcohol are minimized (and yes, we still use xylene! And in case it matters schedule-wise, we use 10% NBFS as well). Thanks in advance. Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. _______________________________________________ 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 ------------------------------ Message: 8 Date: Thu, 8 Nov 2007 11:07:25 -0600 From: "Drew Sally A." Subject: [Histonet] Clone for Prog. Receptor To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" We currently are using the 1A6 clone for our progesterone receptor(PR) IHC. We were wondering how other people felt about rabbit monoclonal PR antibodies. Does anyone have strong preferences or experiences with other PR clones-mouse or rabbit-that they'd like to share? Thank you...! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 ------------------------------ Message: 9 Date: Thu, 08 Nov 2007 12:10:47 -0500 From: John Kiernan Subject: Re: [Histonet] Artefacts To: Toxicology Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Dear Toxicology You need to explain your problem more clearly. Please include histonet@lists.utsouthwestern.edu in your replies. We all want to share our knowledge. What tissue are you staining? How was it fixed? Are you staining dewaxed and hydrated paraffin sections? Cryostat sections? Which haemalum & eosin method did you use? Does your lab have a book with instructions for H&E staining? If not, why not? Are the "faint stained spots on sections" blue, purple, pink or red? Where are these spots? John Kiernan Anatomy, UWO London, Canada. === ----- Original Message ----- From: Toxicology Date: Thursday, November 8, 2007 1:36 Subject: [Histonet] Artefacts To: histonet@lists.utsouthwestern.edu > Dear all, > I am facing a problem in Hematoxylin and Eosin staining. There are > several faint stained spots on sections and because of that its > difficuly to interpret the lesions. I am unable to sort out the > problem. Can any one help me? > > > > > -------------------------Email Disclaimer------------------------ > --------- > ------------------------------ Message: 10 Date: Thu, 8 Nov 2007 17:13:21 -0000 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Artefacts To: "John Kiernan" , "Toxicology" Cc: histonet@lists.utsouthwestern.edu Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE4F@TRFT-EX01.xRothGen.nhs.uk> Content-Type: text/plain; charset="us-ascii" Some of these questions should be answered by the photos, but I cannot find them. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: 08 November 2007 17:11 To: Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Artefacts Dear Toxicology You need to explain your problem more clearly. Please include histonet@lists.utsouthwestern.edu in your replies. We all want to share our knowledge. What tissue are you staining? How was it fixed? Are you staining dewaxed and hydrated paraffin sections? Cryostat sections? Which haemalum & eosin method did you use? Does your lab have a book with instructions for H&E staining? If not, why not? Are the "faint stained spots on sections" blue, purple, pink or red? Where are these spots? John Kiernan Anatomy, UWO London, Canada. === ----- Original Message ----- From: Toxicology Date: Thursday, November 8, 2007 1:36 Subject: [Histonet] Artefacts To: histonet@lists.utsouthwestern.edu > Dear all, > I am facing a problem in Hematoxylin and Eosin staining. There are > several faint stained spots on sections and because of that its > difficuly to interpret the lesions. I am unable to sort out the > problem. Can any one help me? > > > > > -------------------------Email Disclaimer------------------------ > --------- > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Thu, 8 Nov 2007 12:13:22 -0500 (EST) From: "FU,DONGTAO" Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3 To: Histonet@lists.utsouthwestern.edu Message-ID: <813060655.505211194542002919.JavaMail.osg@osgjas04.cns.ufl.edu> Content-Type: text/plain; format=flowed; charset=us-ascii Hello, Can anyone give me some suggestions on my case? I know a lot of people here have a lot of dual staining experiences. I did dual staining on 2 antibodies from different species in the past. They worked well. But using 2 antibodies from same species is my first time try and I met a big problem. Any suggestions I will be very appreciate. Ann On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" wrote: > Hi, all > > Thank you first for giving me some good suggestions on thick-section > question I posted last time. Now I met another problem when I did CD4 > and FOXP3 dual staining using murine fresh frozen spleen. If I did > single staining, both antibodies worked very well. However if I did > dual staining, I could only get good result from the first antibody. > The second one did not work at all(I mean no specific staining). I > used CD4 from BD(rat anti-mouse). FOXP3 I used from ebioscience, also > rat anti-mouse. > I think there might be something wrong with my serum block(I used > normal rat serum block) before I added secondary antibody. Or any > other serum block I need to add to decrease the non-specific binding > which I have not done yet. Does anyone can give me some suggestions > according to my protocol below? How can I get specific staining of the > secondary (primary)antibody? Thank you, > > Below is the simple protocol I used for dual staining: > 1. 2% normal goat serum block 20 min > 2. 1* antibody CD4 1:750 in diluent O/N 4C > 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT > 4. Serum block: 5% normal rat serum 30 min > 5. 1* antibody FOXP3 1:100 in diluent 1h RT > 6. Secondary AF488 Donkey anti-rat in TBS 1h RT > > Use 1xTBS as wash buffer. Before staining, fix tissue in -20C Aceton > for 5 min, then airdry. > > > Ann Dongtao Fu MD Ph.D > Lab Manager > Molecular Pathology core > Dept. of Pathology > University of Florida > 1600 SW Archer Road > Gainesville, FL 32610 > Lab Phone: 352-273-7752 > FAX: 352-273-7753 > Rm: D11-50 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Ann Dongtao Fu MD, Ph.D Lab Manager Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 ------------------------------ Message: 12 Date: Thu, 8 Nov 2007 12:16:04 -0500 From: "Weems, Joyce" Subject: RE: [Histonet] Artefacts To: "Marshall Terry Dr, Consultant Histopathologist" , "John Kiernan" , "Toxicology" Cc: histonet@lists.utsouthwestern.edu Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD32048F41AF@sjhaexc02.sjha.org> Content-Type: text/plain; charset="utf-8" The web site says give it 24 hrs for the photo to post... Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, November 08, 2007 12:13 PM To: John Kiernan; Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Artefacts Some of these questions should be answered by the photos, but I cannot find them. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: 08 November 2007 17:11 To: Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Artefacts Dear Toxicology You need to explain your problem more clearly. Please include histonet@lists.utsouthwestern.edu in your replies. We all want to share our knowledge. What tissue are you staining? How was it fixed? Are you staining dewaxed and hydrated paraffin sections? Cryostat sections? Which haemalum & eosin method did you use? Does your lab have a book with instructions for H&E staining? If not, why not? Are the "faint stained spots on sections" blue, purple, pink or red? Where are these spots? John Kiernan Anatomy, UWO London, Canada. === ----- Original Message ----- From: Toxicology Date: Thursday, November 8, 2007 1:36 Subject: [Histonet] Artefacts To: histonet@lists.utsouthwestern.edu > Dear all, > I am facing a problem in Hematoxylin and Eosin staining. There are > several faint stained spots on sections and because of that its > difficuly to interpret the lesions. I am unable to sort out the > problem. Can any one help me? > > > > > -------------------------Email Disclaimer------------------------ > --------- > _______________________________________________ Histonet mailing list Histonet@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 Joseph's Health System, Inc. ------------------------------ Message: 13 Date: Thu, 8 Nov 2007 12:20:12 -0500 From: "Weems, Joyce" Subject: RE: [Histonet] B5 substitute To: Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD32048F41B0@sjhaexc02.sjha.org> Content-Type: text/plain; charset="utf-8" We use B-Plus from BBC. http://www.bbcus.com/website_search.html It takes a long time to get used to, especially when they have to compare to previous mercury fixed cases, but it can be done! Good luck, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne Mauger Sent: Thursday, November 08, 2007 11:51 AM To: gvdobbin@ihis.org; Histonet@lists.utsouthwestern.edu Subject: [Histonet] B5 substitute Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ 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 Joseph's Health System, Inc. ------------------------------ Message: 14 Date: Thu, 8 Nov 2007 09:24:17 -0800 (PST) From: Matt Bancroft Subject: Re: [Histonet] B5 substitute To: Joanne Mauger , gvdobbin@ihis.org, Histonet@lists.utsouthwestern.edu Message-ID: <704342.79971.qm@web63407.mail.re1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 B-Plus Joanne Mauger wrote: Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ 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 ------------------------------ Message: 15 Date: Thu, 8 Nov 2007 17:27:01 -0000 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Artefacts To: "Weems, Joyce" , "John Kiernan" , "Toxicology" Cc: histonet@lists.utsouthwestern.edu Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE50@TRFT-EX01.xRothGen.nhs.uk> Content-Type: text/plain; charset="us-ascii" Yes, I know. Posters can read that too and wait until the photos appear before they post the question. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: 08 November 2007 17:16 To: Marshall Terry Dr, Consultant Histopathologist; John Kiernan; Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Artefacts The web site says give it 24 hrs for the photo to post... Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Thursday, November 08, 2007 12:13 PM To: John Kiernan; Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Artefacts Some of these questions should be answered by the photos, but I cannot find them. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: 08 November 2007 17:11 To: Toxicology Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Artefacts Dear Toxicology You need to explain your problem more clearly. Please include histonet@lists.utsouthwestern.edu in your replies. We all want to share our knowledge. What tissue are you staining? How was it fixed? Are you staining dewaxed and hydrated paraffin sections? Cryostat sections? Which haemalum & eosin method did you use? Does your lab have a book with instructions for H&E staining? If not, why not? Are the "faint stained spots on sections" blue, purple, pink or red? Where are these spots? John Kiernan Anatomy, UWO London, Canada. === ----- Original Message ----- From: Toxicology Date: Thursday, November 8, 2007 1:36 Subject: [Histonet] Artefacts To: histonet@lists.utsouthwestern.edu > Dear all, > I am facing a problem in Hematoxylin and Eosin staining. There are > several faint stained spots on sections and because of that its > difficuly to interpret the lesions. I am unable to sort out the > problem. Can any one help me? > > > > > -------------------------Email Disclaimer------------------------ > --------- > _______________________________________________ Histonet mailing list Histonet@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 Joseph's Health System, Inc. ------------------------------ Message: 16 Date: Thu, 08 Nov 2007 12:36:02 -0500 From: "Richard Cartun" Subject: Re: [Histonet] B5 substitute To: "Joanne Mauger" ,, Message-ID: <473302B20200007700009021@gwmail4.harthosp.org> Content-Type: text/plain; charset=US-ASCII Formalin (fix well and cut it thin). 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 >>> "Joanne Mauger" 11/08/07 11:50 AM >>> Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ 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: 17 Date: Thu, 08 Nov 2007 12:40:02 -0500 From: "Richard Cartun" Subject: Re: [Histonet] Clone for Prog. Receptor To: "Histonet" , "Drew Sally A." Message-ID: <473303A20200007700009027@gwmail4.harthosp.org> Content-Type: text/plain; charset=US-ASCII We have used clone PgR636 (from Dako) for several years now with excellent results. 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 >>> "Drew Sally A." 11/08/07 12:07 PM >>> We currently are using the 1A6 clone for our progesterone receptor(PR) IHC. We were wondering how other people felt about rabbit monoclonal PR antibodies. Does anyone have strong preferences or experiences with other PR clones-mouse or rabbit-that they'd like to share? Thank you...! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 _______________________________________________ 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: 18 Date: Thu, 8 Nov 2007 09:45:24 -0800 From: "Tarango, Mark" Subject: RE: [Histonet] problem in dual staining of CD4 and FOXP3 To: "FU,DONGTAO" , Histonet@lists.utsouthwestern.edu Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F71@NVCIEXCH02.NVCI.org> Content-Type: text/plain; charset=us-ascii Looking at T-regulatory cells, huh? Are you trying to stain both in the same color? I know Foxp3 is nuclear and CD4 is on the cell membrane, but two colors might be better. Your protocol looks incomplete. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of FU,DONGTAO Sent: Thursday, November 08, 2007 9:13 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3 Hello, Can anyone give me some suggestions on my case? I know a lot of people here have a lot of dual staining experiences. I did dual staining on 2 antibodies from different species in the past. They worked well. But using 2 antibodies from same species is my first time try and I met a big problem. Any suggestions I will be very appreciate. Ann On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" wrote: > Hi, all > > Thank you first for giving me some good suggestions on > thick-section question I posted last time. Now I met another > problem when I did CD4 and FOXP3 dual staining using murine fresh > frozen spleen. If I did single staining, both antibodies worked > very well. However if I did dual staining, I could only get good > result from the first antibody. The second one did not work at > all(I mean no specific staining). I used CD4 from BD(rat > anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. > I think there might be something wrong with my serum block(I used > normal rat serum block) before I added secondary antibody. Or any > other serum block I need to add to decrease the non-specific > binding which I have not done yet. Does anyone can give me some > suggestions according to my protocol below? How can I get > specific staining of the secondary (primary)antibody? Thank you, > > Below is the simple protocol I used for dual staining: > 1. 2% normal goat serum block 20 min > 2. 1* antibody CD4 1:750 in diluent O/N 4C > 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT > 4. Serum block: 5% normal rat serum 30 min > 5. 1* antibody FOXP3 1:100 in diluent 1h RT > 6. Secondary AF488 Donkey anti-rat in TBS 1h RT > > Use 1xTBS as wash buffer. Before staining, fix tissue in -20C > Aceton for 5 min, then airdry. > > > Ann Dongtao Fu MD Ph.D > Lab Manager > Molecular Pathology core > Dept. of Pathology > University of Florida > 1600 SW Archer Road > Gainesville, FL 32610 > Lab Phone: 352-273-7752 > FAX: 352-273-7753 > Rm: D11-50 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Ann Dongtao Fu MD, Ph.D Lab Manager Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ---------------------------------------------------------------------------- -- CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================ == ------------------------------ Message: 19 Date: Thu, 8 Nov 2007 09:47:54 -0800 From: "Tarango, Mark" Subject: RE: [Histonet] B5 substitute To: "Joanne Mauger" , gvdobbin@ihis.org, Histonet@lists.utsouthwestern.edu Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F72@NVCIEXCH02.NVCI.org> Content-Type: text/plain; charset=us-ascii Why not just use 10% NBF? It's great if you allow enough time for fixation. Make sure your bone marrows are fixed well BEFORE decalcifying and you should have no problems. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joanne Mauger Sent: Thursday, November 08, 2007 8:51 AM To: gvdobbin@ihis.org; Histonet@lists.utsouthwestern.edu Subject: [Histonet] B5 substitute Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ---------------------------------------------------------------------------- -- CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================ == ------------------------------ Message: 20 Date: Thu, 8 Nov 2007 09:58:22 -0800 (PST) From: Steven Coakley Subject: [Histonet] HT position wanted To: Histonet@lists.utsouthwestern.edu Message-ID: <621094.56475.qm@web38209.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I'm looking for an HT position in the Madison, WI to Rockford, Ill area. Contract in Milwaukee, WI. Thanks, Steve __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 21 Date: Thu, 8 Nov 2007 12:59:10 -0500 (EST) From: "FU,DONGTAO" Subject: RE: [Histonet] problem in dual staining of CD4 and FOXP3 To: Histonet@lists.utsouthwestern.edu Message-ID: <1211976397.304891194544750373.JavaMail.osg@osgjas03.cns.ufl.edu> Content-Type: text/plain; format=flowed; charset=us-ascii Hi, Mark, Yes, I am looking at T-regulatory cells of mouse. For CD4, I used AF594(red color), for FOXP3 I used AF488(green color). The problem for me is for the second primary antibody(or maybe I should say for the secondary color) of dual staining, it never worked. I do not know which serum I should use to block the non-specific staining of the secondary antibody. If you think my protocol is imcomplete, could give me some suggestions? Many thanks, Ann On Thu Nov 08 12:45:24 EST 2007, "Tarango, Mark" wrote: > Looking at T-regulatory cells, huh? Are you trying to stain both > in the > same color? I know Foxp3 is nuclear and CD4 is on the cell > membrane, > but two colors might be better. Your protocol looks incomplete. > Mark Adam Tarango HT(ASCP) > Histology & IHC Supervisor > Nevada Cancer Institute > One Breakthrough Way > Las Vegas, NV 89135 > Direct Line (702) 822-5112 > Treo (702) 759-9229 > Fax (702) 939-7663 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > FU,DONGTAO > Sent: Thursday, November 08, 2007 9:13 AM > To: Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] problem in dual staining of CD4 and FOXP3 > > Hello, > > Can anyone give me some suggestions on my case? I know a lot of > people here have a lot of dual staining experiences. I did dual > staining on 2 antibodies from different species in the past. They > worked well. But using 2 antibodies from same species is my first > time try and I met a big problem. Any suggestions I will be very > appreciate. > > Ann > > > On Thu Nov 08 08:38:39 EST 2007, "FU,DONGTAO" > wrote: > >> Hi, all >> >> Thank you first for giving me some good suggestions on >> thick-section question I posted last time. Now I met another >> problem when I did CD4 and FOXP3 dual staining using murine >> fresh frozen spleen. If I did single staining, both antibodies >> worked very well. However if I did dual staining, I could only >> get good result from the first antibody. The second one did not >> work at all(I mean no specific staining). I used CD4 from BD(rat >> anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. >> I think there might be something wrong with my serum block(I >> used normal rat serum block) before I added secondary antibody. >> Or any other serum block I need to add to decrease the >> non-specific binding which I have not done yet. Does anyone can >> give me some suggestions according to my protocol below? How can >> I get specific staining of the secondary (primary)antibody? >> Thank you, >> >> Below is the simple protocol I used for dual staining: >> 1. 2% normal goat serum block 20 min >> 2. 1* antibody CD4 1:750 in diluent O/N 4C >> 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT >> 4. Serum block: 5% normal rat serum 30 min >> 5. 1* antibody FOXP3 1:100 in diluent 1h RT >> 6. Secondary AF488 Donkey anti-rat in TBS 1h RT >> >> Use 1xTBS as wash buffer. Before staining, fix tissue in -20C >> Aceton for 5 min, then airdry. >> >> >> Ann Dongtao Fu MD Ph.D >> Lab Manager >> Molecular Pathology core >> Dept. of Pathology >> University of Florida >> 1600 SW Archer Road >> Gainesville, FL 32610 >> Lab Phone: 352-273-7752 >> FAX: 352-273-7753 >> Rm: D11-50 >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > > > Ann Dongtao Fu MD, Ph.D > Lab Manager > Dept. of Pathology > Lab phone: 352-273-7752 > Lab FAX: 352-273-7755 > Lab address: D11-50 > PO Box: 100275 > 1600 SW Archer Road > University of Flodrida > Gainesville, FL 32610 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > "EMF " made the following annotations. > ---------------------------------------------------------------------------- -- > CONFIDENTIALITY NOTICE: This e-mail message, including any > attachments, is for the sole use of the intended recipient(s) and > may contain confidential, proprietary, and/or privileged > information protected by law. If you are not the intended > recipient, you may not use, copy, or distribute this e-mail > message or its attachments. If you believe you have received this > e-mail message in error, please contact the sender by reply > e-mail and destroy all copies of the original message > ============================================================================ == > > > Ann Dongtao Fu MD, Ph.D Lab Manager Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 11 **************************************** From arnie.jimenez <@t> vel-lab.com Thu Nov 8 16:21:12 2007 From: arnie.jimenez <@t> vel-lab.com (arnie.jimenez@vel-lab.com) Date: Thu Nov 8 16:23:29 2007 Subject: [Histonet] Re: Away from Office Message-ID: <20071108222112.31568.qmail@plesk5.hostgator.com> Thank you for contacting Vel-Lab Research. We will be out of the lab starting Nov. 7th and will return on Nov. 12th. We will check e-mail regularly but will be unable to respond. Please do not mail any packages to our lab until Nov. 12th. Regards, Arnie Jimenez Vel-Lab Research From mtarango <@t> nvcancer.org Thu Nov 8 17:35:13 2007 From: mtarango <@t> nvcancer.org (Tarango, Mark) Date: Thu Nov 8 17:41:39 2007 Subject: [Histonet] Dendritic cells in mouse FFPE tissue In-Reply-To: <07979E76B0869D4E8C9FE4AA9FC0657804668BC5@srvex03.phsabc.ehcnet.ca> Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F76@NVCIEXCH02.NVCI.org> What kind of dendritic cells? I have a research antibody that I SWEAR is marking the nuclei of dendritic cells. The researcher who made the antibody claims they're stem cells though. What do I know? Those langerhans kinds of dendritic cells should be CD1a positive and BIOCARE has an antibody that works on paraffin. You might try CD11c too. For plasmacytoid dendritic cells or follicular center/germinal center dendritic cells I don't know of any single marker that is great and won't mark anything aside from the dendritic cells. You don't want any cross-reactivity with macrophages, so I guess CD68 ain't an option... Hmmm.... Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Milne, Katy Sent: Thursday, November 08, 2007 1:22 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dendritic cells in mouse FFPE tissue Ok, I know I'm on another one of my wild goose chases again but is anyone successfully working with an antibody that detects dendritic cells in mouse FFPE tissue (no, frozen is not an option at this point)? It needs to be something specific for dendritic cells, i.e. no cross-reactivity with macrophages etc. Thanks, Katy Milne Deeley Research Centre _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== From lpwenk <@t> sbcglobal.net Thu Nov 8 19:04:12 2007 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Thu Nov 8 19:04:46 2007 Subject: [Histonet] B5 substitute In-Reply-To: Message-ID: <002701c8226c$71801190$0202a8c0@HPPav2> Couple of hints with any B5 substitute (which are usually zinc formalin), from a research project 2 of our students did on various brands of zinc formalin at different time intervals - 1. If you are going to place zinc formalin in the tissue processor, use zinc sulfate, not zinc chloride. When zinc chloride cross-links with protein, it releases hydrochloric acid, which can chew up the retort chamber of the processor (hint from a Journal of Histotechnology article a lot of years ago, one of which authors was Liz Chlipala, who contributes to Histonet). 2. However long you are used to fixing the tissue in B5, multiply that number by 1.5, and that will be about right for the minimum time in zinc formalin. Zinc binds slower than mercury - plain simple chemistry. So if you were doing bone marrow biopsies for 2 hour in B5, 2 x 1.5 = 3 hours in zinc formalin. If the lymph nodes were taking 4 hours in B5, try 4 x 1.5 = 6 hours in zinc formalin. The first time we tried this experiment with different brands of zinc formalin, we kept the times the same as B5. All the tissues had awful looking nuclei. So we dropped the idea of switching. A couple of years later, we tried the experiment again, with time intervals of something like 1, 2, 3, 4, 6, 8 and 24 hours. And we did it on bone marrow biopsies and on lymph nodes. That's when we came up with the "multiply it by 1.5" factor. It seemed pretty universal, regardless of which company. After coming up with the best time, we conveyed the information to the med techs who go up on the bone marrow biopsies. Most of the biopsies looked fine, but every once in a while, one would have nuclei that looked like a bad alcian blue on goblet cells. Turns out, one of the med techs didn't like the change in routine - "but I've always done it for 2 hours" and was refusing to change to 3 hours, until we showed her other tech's well fixed zinc formalin bone marrows and her zinc formalin-fixed bone marrows, and asked her which one she would like the hematopathologist to diagnosis from, if this was her daughter's biopsy for possible leukemia. No problems after that. 3. You may need 2 different zinc formalins. We found one worked better for tissues that were going to go into acid for decalcification, and a different one worked better for all other tissues (tumors, lymph nodes). Sorry, I just don't remember which companies we liked. I'd have to dig out the research papers. 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 Joanne Mauger Sent: Thursday, November 08, 2007 11:51 AM To: gvdobbin@ihis.org; Histonet@lists.utsouthwestern.edu Subject: [Histonet] B5 substitute Hi , What are you using to replace B5 fixative with mercury? I want the best for hematopoetic tissues, and for immunostains. Thanks, Jo Mauger _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Tony_Reilly <@t> health.qld.gov.au Thu Nov 8 21:56:10 2007 From: Tony_Reilly <@t> health.qld.gov.au (Anthony Reilly) Date: Thu Nov 8 22:18:15 2007 Subject: [Histonet] Short processing schedule-suggestions Message-ID: Hi Greg We do a lot of short cycling of biopsies and until recently were using an older model VIP circa 1990. I found that a 2 hour cycle was adequate for your standard sized gut biopsies, however longer was required for the polypoid lesions which can come in all shapes and sizes. So in short for standard sized biopsies (up to 4mm diameter) 2 hours is adequate but if you wanted to be conservative 3 or 4 hours would not be too long. regards Tony Reilly Chief Scientist Anatomical Pathology Pathology Queensland Prince Charles Hospital Rode Rd Chermside Q 4032 Australia Ph: 07 3139 4543 Fax: 07 3193 4546 tony_reilly@health.qld.gov.au >>> "Greg Dobbin" 11/09/07 12:59 am >>> Hello All, I would like to get your suggestions for a processing schedule for small biopsies (cores, GI's, other endoscopics). I want to process these little guys separate from everything else to hopefully improve section quality and perhaps staining quality. I don't want the absolute shortest possible schedule. We have a newer VIP5 for our main processor (doing everything on one schedule currently) and a much older VIP sitting idle as a backup that I plan to start using for the biopsies. We process overnight so I have more than enough time. I will just delay the biopsy processing so that both processors are finished at essentially the same time. What I need is a shorter schedule for the biopsies so that the detrimental affects of xylene and alcohol are minimized (and yes, we still use xylene! And in case it matters schedule-wise, we use 10% NBFS as well). Thanks in advance. Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. 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The information contained in this email, including any attachment sent with it, may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone collect on Australia +61 1800 198 175 or by return email. You should also delete this email, and any copies, from your computer system network and destroy any hard copies produced. If not an intended recipient of this email, you must not copy, distribute or take any action(s) that relies on it; any form of disclosure, modification, distribution and/or publication of this email is also prohibited. Although Queensland Health takes all reasonable steps to ensure this email does not contain malicious software, Queensland Health does not accept responsibility for the consequences if any person's computer inadvertently suffers any disruption to services, loss of information, harm or is infected with a virus, other malicious computer programme or code that may occur as a consequence of receiving this email. Unless stated otherwise, this email represents only the views of the sender and not the views of the Queensland Government. **************************************************************** From sitiariza <@t> yahoo.com Thu Nov 8 23:34:55 2007 From: sitiariza <@t> yahoo.com (siti ariza) Date: Thu Nov 8 23:35:00 2007 Subject: [Histonet] studies on digestive enzymes of fish laravae Message-ID: <576275.53813.qm@web33610.mail.mud.yahoo.com> Hello, i'm currently doing my M.Sc and working on histochemistry of fish larvae. For this i will be looking at digestive enzymes of early development of fish larvae. i'm not sure about the method of preparation before incubation sections. in this study, i'm looking for acid phosphatase, alkali phosphatase and lipase enzymes. i hope someone could give me some information on this. thanks Siti Ariza Aripin, Student, Universiti Malaysia Terengganu, Malaysia. __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Kemlo.Rogerson <@t> waht.swest.nhs.uk Fri Nov 9 02:03:40 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Fri Nov 9 02:03:47 2007 Subject: [Histonet] Hematoxylin to dark on some slides Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F132@wahtntex2.waht.swest.nhs.uk> You mean you stain at the same time and those cut at night are staining darker with haematoxylin than those cut during the day. If you are staining those cut at night, also at night, maybe you can't see the clock? Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; No snowflake in an avalanche ever feels responsible. --George Burns This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From c.m.vanderloos <@t> amc.uva.nl Fri Nov 9 02:08:07 2007 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Nov 9 02:47:13 2007 Subject: [Histonet] RE: problem in dual staining of CD4 and FOXP3 Message-ID: <20508a20ccf6.20ccf620508a@amc.uva.nl> Hello Ann, Looking at your double staining protocol I noticed that the normal rat serum blocking is not blocking but making the cross-reaction problem even worse. Normal rat serum will bind at randomly all over the tissue section and your last step 6 (donkey anti-rat/AF488) will bind to it. Therefore I am sure that your whole tissue section is glowing bright green! Our research group have recently published a paper in showing CD4/FOXP3 double staining in human tissues, but we used enzymatic labels here (De Boer et al., JHC 2007, 55:891-898). The use of enzymatic labels allows a relatively easy sequential double staining procedure. However, immuno fluorescence double staining with two unlabeled primaries of the same species is not easy. Most promising is a kind of sequential approach starting with a very high dilution of the first primary detected by a super sensitive fluorescent tyramide amplification method. The second primary antibody is simply detected by a 2-step procedure. No blocking step in between! The rationale behind this, is the very high dilution of the first primary that is not detected by the second detection system. See Brouns et al., JHC 2002, 50:575-582. You have to test a dilution series of the first primary followed by the tyramide-fluo1 detection, then omitting the second primary and applying the final anti-rat/fluo2 step. From the dilution series you have find a dilution of your first primary that is showing a good signal but that is not picked up by the second detection system. Good luck! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Thu, 8 Nov 2007 08:38:39 -0500 (EST) From: "FU,DONGTAO" Subject: [Histonet] problem in dual staining of CD4 and FOXP3 To: histonet@lists.utsouthwestern.edu Hi, all Thank you first for giving me some good suggestions on thick-section question I posted last time. Now I met another problem when I did CD4 and FOXP3 dual staining using murine fresh frozen spleen. If I did single staining, both antibodies worked very well. However if I did dual staining, I could only get good result from the first antibody. The second one did not work at all(I mean no specific staining). I used CD4 from BD(rat anti-mouse). FOXP3 I used from ebioscience, also rat anti-mouse. I think there might be something wrong with my serum block(I used normal rat serum block) before I added secondary antibody. Or any other serum block I need to add to decrease the non-specific binding which I have not done ye! t. Does a From sbledsoe <@t> iupui.edu Fri Nov 9 06:37:03 2007 From: sbledsoe <@t> iupui.edu (Sharon Bledsoe) Date: Fri Nov 9 06:37:21 2007 Subject: [Histonet] Hematoxylin to dark on some slides In-Reply-To: <86ADE4EB583CE64799A9924684A0FBBF0222F132@wahtntex2.waht.swest.nhs.uk> References: <86ADE4EB583CE64799A9924684A0FBBF0222F132@wahtntex2.waht.swest.nhs.uk> Message-ID: Or the slice thickness At 8:03 AM +0000 11/9/07, Kemlo Rogerson wrote: >You mean you stain at the same time and those cut at night are staining >darker with haematoxylin than those cut during the day. If you are >staining those cut at night, also at night, maybe you can't see the >clock? > >Kemlo Rogerson >Pathology Manager >DD 01934 647057 or extension 3311 >Mob 07749 754194; Pager 07659 597107; >No snowflake in an avalanche ever feels responsible. --George Burns > >This e-mail is confidential and privileged. If you are not the intended >recipient please accept my 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 me that this message has gone astray before deleting it. >Thank you for your co-operation > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Farnana <@t> nehealth.com Fri Nov 9 06:50:28 2007 From: Farnana <@t> nehealth.com (Amy Farnan) Date: Fri Nov 9 06:50:53 2007 Subject: [Histonet] biopsy run schedule Message-ID: <47341144.26ED.00D9.0@nehealth.com> Good morning, I currently have a VIP300 and a VIP5. I run my routines on the 5 and run a biopsy run on the 300. My biopsy run is 15 min. in each solution and they come out great. I also have it delayed so they come off at the same time. I use the following solutions: 2- formalin 1 - 70 % - found Penfix tends to over fix and make them drier. 1- 80% 2- 95% 2- 100% 2- xylene 3- paraffin I hope this helps...good luck Disclaimer: The information in this message is confidential. If you are not the intended recipient, do not disclose, copy, or distribute this message, and please immediately contact the sender. From cmiller <@t> physlab.com Fri Nov 9 07:27:56 2007 From: cmiller <@t> physlab.com (Cheri Miller) Date: Fri Nov 9 07:30:02 2007 Subject: [Histonet] Hematoxylin to dark on some slides In-Reply-To: References: <86ADE4EB583CE64799A9924684A0FBBF0222F132@wahtntex2.waht.swest.nhs.uk> Message-ID: <008701c822d4$572039c0$3402a8c0@plab.local> Who is cutting them? I know I can have 2 techs cutting at the same microns, same tissue and one's sections will always be thicker regardless of which microtome they are cutting on. I have come to the conclusion its all in the technique of each individual tech. Cheri Cheri Miller HT ASCP Histology Supervisor Physicians Laboratory Services, Inc. Omaha, NE 68117 402 738 5052 PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Bledsoe Sent: Friday, November 09, 2007 6:37 AM To: Kemlo Rogerson; Dawn Bugge; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Hematoxylin to dark on some slides Or the slice thickness At 8:03 AM +0000 11/9/07, Kemlo Rogerson wrote: >You mean you stain at the same time and those cut at night are staining >darker with haematoxylin than those cut during the day. If you are >staining those cut at night, also at night, maybe you can't see the >clock? > >Kemlo Rogerson >Pathology Manager >DD 01934 647057 or extension 3311 >Mob 07749 754194; Pager 07659 597107; >No snowflake in an avalanche ever feels responsible. --George Burns > >This e-mail is confidential and privileged. If you are not the intended >recipient please accept my 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 me that this message has gone astray before deleting it. >Thank you for your co-operation > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. From Kemlo.Rogerson <@t> waht.swest.nhs.uk Fri Nov 9 07:31:32 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Fri Nov 9 07:31:39 2007 Subject: [Histonet] Hematoxylin to dark on some slides Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F144@wahtntex2.waht.swest.nhs.uk> Why would they cut thicker sections at night? If they couldn't see the thickness knob and changed it they would sometimes have thick and thin sections. Is it warmer at night? Do the night people stain using a different procedure? Or the slice thickness Sharon At 8:03 AM +0000 11/9/07, Kemlo Rogerson wrote: >You mean you stain at the same time and those cut at night are staining >darker with haematoxylin than those cut during the day. If you are >staining those cut at night, also at night, maybe you can't see the >clock? > Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; Nonviolence means avoiding not only external physical violence but also internal violence of spirit. You not only refuse to shoot a man, but you refuse to hate him. --Martin Luther King Jr. This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From Beatrice.Debrosse-Serra <@t> pfizer.com Fri Nov 9 08:20:43 2007 From: Beatrice.Debrosse-Serra <@t> pfizer.com (Debrosse-Serra, Beatrice) Date: Fri Nov 9 08:21:11 2007 Subject: [Histonet] Pfizer, La Jolla, CA, Histology opening Message-ID: <8404DFBED5207B4B8E5EEF4332CEEA5304BEFB52@lajamrexm01.amer.pfizer.com> Hi Histonetters, We have an opening for a histologist here at Pfizer in beautiful La Jolla, CA. Below you find the job description. If you are interested, please forward your r?sum? to me. Thanks, Bea Responsibilities: The ideal candidate will have a high degree of proficiency in routine histology. The successful individual will be responsible for the technical procedures utilized in the preparation of microscopic slides including tissue processing, paraffin block sectioning, and H&E staining. Computer literacy is required. Main responsibilities will be production of high quality slides from frozen or fixed tissues. This will include trimming, embedding, processing, sectioning using a microtome/cryostat, and staining with H&E or other stains. Conduct slide reviews for quality control of tissue sections. Produce a quality product in a timely manner. A microtomy sectioning rate of approximately 60 blocks/day with a <3% recut rate is expected. Experience in immunohistochemistry and in situ hybridization is desirable but not essential. Other tasks may include file maintenance; other instrument and equipment maintenance; quality control checks; and related laboratory tasks. Qualifications: The successful candidate will have a Bachelor/Master degree in biology or other life sciences, be certified as a Histotechnician (HT) or Histotechnologist (HTL) and have at least 2 years of experience in a histology setting. Experience in working in a GLP environment is desirable. Proficiency in necropsy techniques or a willingness to be trained in this area is desirable. Strong written and oral communication skills are essential as the selected individual will be required to contribute to written SOP and/or in group meetings. The ability to develop effective working relationships in cross-functional teams, strong interpersonal skills, and strong troubleshooting skills are also essential. Beatrice DeBrosse-Serra Pathology Scientist Pfizer Global Research & Development CB4, 2150 10646 Science Center Drive San Diego, CA 92121 Phone# 858-622-5986 Fax# 858-678-8290 From Luis.Chiriboga <@t> med.nyu.edu Fri Nov 9 08:38:22 2007 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Nov 9 08:41:00 2007 Subject: [Histonet] TGIF Message-ID: *_NEW HOSPITAL WING_* When a panel of doctors was asked to vote on adding a new wing to their hospital, the Allergists voted to scratch it, and the Dermatologists advised not to make any rash moves. The Gastroenterologists had sort of a gut feeling about it, but the Neurologists thought the administration had a lot of nerve, and the Obstetricians felt they were all laboring under a misconception The Ophthalmologists considered the idea shortsighted; the Pathologists yelled, 'Over my dead body', while the Pediatricians said, 'Oh, grow up!' The Psychiatrists thought the whole idea was madness, the Radiologists could see right through it, and the Surgeons decided to wash their hands of the whole thing. The Internists thought it was a bitter pill to swallow, and the Plastic Surgeons said, 'This puts a whole new face on the matter.' The Podiatrists thought it was a step forward, but the Urologists felt the scheme wouldn't hold water. The Anesthesiologists thought the whole idea was a gas, and the Cardiologists didn't have the heart to say no. In the end, the Proctologists left the decision up to some a....... in administration. From Terry.Marshall <@t> rothgen.nhs.uk Fri Nov 9 10:13:47 2007 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Nov 9 10:13:53 2007 Subject: [Histonet] Fwd: Artefacts Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE5B@TRFT-EX01.xRothGen.nhs.uk> The pictures are still not there. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Toxicology Sent: 08 November 2007 06:50 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fwd: Artefacts Dear all, I am facing a problem in Hematoxylin and Eosin staining. There are several faint stained spots on sections and because of that its difficuly to interpret the lesions. I am unable to sort out the problem. Can any one help me? I have posted a picture to www.histonet.org. named artefact. -----Original Message----- From: "Toxicology" To: histonet@lists.utsouthwestern.edu Date: Thu, 08 Nov 2007 12:11:41 +0530 Subject: Artefacts Dear all, I am facing a problem in Hematoxylin and Eosin staining. There are several faint stained spots on sections and because of that its difficuly to interpret the lesions. I am unable to sort out the problem. Can any one help me? -------------------------Email Disclaimer--------------------------------- This e-mail may contain confidential and/or privileged information. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janci.Wellborn <@t> stlukes-stl.com Fri Nov 9 10:20:24 2007 From: Janci.Wellborn <@t> stlukes-stl.com (Wellborn, Janci R) Date: Fri Nov 9 10:20:30 2007 Subject: [Histonet] biopsy run schedule In-Reply-To: <47341144.26ED.00D9.0@nehealth.com> Message-ID: <42DC3293408E094499C68DDDD45CA08E35CA73@W3CEXCHANGE2.slh.stlukes.com> Janci R Wellborn, BS, BSeD, HTL (ASCP) Anatomic Pathology Supervisor St. Luke's Hospital 232 South Woods Mill Road Chesterfield, Missouri 63017 (314) 205-6228 janci.wellborn@stlukes-stl.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amy Farnan Sent: Friday, November 09, 2007 6:50 AM To: gvdobbin@ihis.org; histonet@lists.utsouthwestern.edu Subject: [Histonet] biopsy run schedule Good morning, I currently have a VIP300 and a VIP5. I run my routines on the 5 and run a biopsy run on the 300. My biopsy run is 15 min. in each solution and they come out great. I also have it delayed so they come off at the same time. I use the following solutions: 2- formalin 1 - 70 % - found Penfix tends to over fix and make them drier. 1- 80% 2- 95% 2- 100% 2- xylene 3- paraffin I hope this helps...good luck Disclaimer: The information in this message is confidential. If you are not the intended recipient, do not disclose, copy, or distribute this message, and please immediately contact the sender. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: The contents of this e-mail, including any attachments, contain information which may be confidential, legally privileged, proprietary in nature, or otherwise protected by law from disclosure, and is solely for the use of the intended recipient(s). If you are not the intended recipient, any use, disclosure or copying of this e-mail, including any attachments, is unauthorized and strictly prohibited. If you have received this e-mail in error, please notify us via return e-mail and immediately delete all copies of it from your system. Any opinions either expressed or implied in this e-mail and all attachments, are those of its author only, and do not necessarily reflect those of St. Luke's Hospital. From amylee779 <@t> yahoo.com Fri Nov 9 11:04:30 2007 From: amylee779 <@t> yahoo.com (Amy Lee) Date: Fri Nov 9 11:04:45 2007 Subject: [Histonet] Go frozen after 10% NBF and 70% EtOH? Message-ID: <995704.72384.qm@web38003.mail.mud.yahoo.com> Hello histonetters, We received a few whole rat brains in 70% EtOH after perfused with 10% NBF. Now they changed mind want go for frozen. The want Cresyl Violet stain. Is it doable? How? Thanks in advance, Amy __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From johnzhan <@t> yahoo.com Fri Nov 9 11:26:34 2007 From: johnzhan <@t> yahoo.com (J. Zhan) Date: Fri Nov 9 11:26:37 2007 Subject: [Histonet] I need a chance of clinical practicum Message-ID: <333264.70596.qm@web31715.mail.mud.yahoo.com> Hi, Histonetters: Because of my foreign training and years working background in histology, histochemistry and IHC, before I am eligible to take ASCP exam I need at least one-year clinical experience under guidance of a ASCP-certified supervisor/pathologist in US. Is there any lab to be able to accept me as apprentice/intern without payment? Meanwhile, I am going to join a on-line training program as well, which can shorten this training, but it is necessary to have a qualified supervisor for me during this training, too. Please help me and save my American dream. Your advice is welcome! You can contact me at 626-377-3153 or email me to johnzhan@yahoo.com. Thank you for your attention! Sincerely, John Zhan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From jdhisto <@t> yahoo.com Fri Nov 9 11:55:55 2007 From: jdhisto <@t> yahoo.com (Jonathan De La Rosa) Date: Fri Nov 9 11:55:59 2007 Subject: [Histonet] Medical technology chat room? Message-ID: <999090.90745.qm@web30301.mail.mud.yahoo.com> Hello, Does anyone know of a "Medical Technology" chat room like this one? Thanks in advance... Jonathan D. From arnie.jimenez <@t> vel-lab.com Fri Nov 9 12:04:13 2007 From: arnie.jimenez <@t> vel-lab.com (arnie.jimenez@vel-lab.com) Date: Fri Nov 9 12:06:40 2007 Subject: [Histonet] Re: Away from Office Message-ID: <20071109180413.31627.qmail@plesk5.hostgator.com> Thank you for contacting Vel-Lab Research. We will be out of the lab starting Nov. 7th and will return on Nov. 12th. We will check e-mail regularly but will be unable to respond. Please do not mail any packages to our lab until Nov. 12th. Regards, Arnie Jimenez Vel-Lab Research From gvdobbin <@t> ihis.org Fri Nov 9 12:06:51 2007 From: gvdobbin <@t> ihis.org (Greg Dobbin) Date: Fri Nov 9 12:07:11 2007 Subject: [Histonet] Medical technology chat room? Message-ID: MEDLAB-L run out of the U. of Alberta (if it is still running??) Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> Jonathan De La Rosa 11/9/2007 1:55:55 PM >>> Hello, Does anyone know of a "Medical Technology" chat room like this one? Thanks in advance... Jonathan D. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. From SBaldwin <@t> compucyte.com Fri Nov 9 16:11:35 2007 From: SBaldwin <@t> compucyte.com (Scott Baldwin) Date: Fri Nov 9 16:11:31 2007 Subject: [Histonet] Duke University Symposium Message-ID: To All Histonetters: There will be a 1/2 day symposium at Duke University on December 6th from 1:30-5:30 at the Ground Floor Auditorium Hock Plaza 2424 Erwin Road Durham, NC 27705 Directions: http://medicalphysics.duke.edu/hock/index.html Amongst the presenters will be David L. Krull, Senior Scientist, Molecular & Ultrastructural Pathology, GSK Safety Assessment, who recently presented at the National Society of Histotechnology Convention in Denver this month on "Quantification of Qdot Labelled Pancreas Using Laser Scanning Cytometry". There will be topics of interest in the area of histopathology and automated tissue analysis. Hopefully those of you that are interested can make it to the symposium. Scott Baldwin (CompuCyte Corporation) From STapper <@t> smdc.org Fri Nov 9 16:45:26 2007 From: STapper <@t> smdc.org (Tapper, Sheila J.) Date: Fri Nov 9 16:46:02 2007 Subject: [Histonet] HT position available in Duluth MN Message-ID: SMDC Clinical Laboratories has a full time position available for an HT or eligible candidate. Our Laboratory offers the opportunity to work with state of the art equipment in a supportive environment. Our pathology group is forward thinking and appreciative of the techs. We of course offer an excellent salary, paid time off, and retirement benefits. What sets us apart are the people you work with, and location, location, location. We don't have the ocean nearby - but we are on the beautiful shores of Lake Superior. We can't deliver a temperate climate but there is nothing more beautiful than the change of seasons in Northern Minnesota. Duluth is not a large metropolis, but we have our own Symphony and Ballet Company. The community theater is very active, and traveling Broadway shows are presented every year in the DECC auditorium. Duluth is home to the Bluesfest - an annual outdoor concert fest held at the beautiful Bayfront Park, right next to the water. For those that need to get away from it all - you can! The Boundary Waters are about 110 miles away, and Minneapolis/St. Paul is only 150 miles away. If you like park-like atmospheres, we have fabulous green space within the city limits that often offer more wildlife than you may want to see - bear, deer and occasionally moose on our 150 miles of trails located in the city. An annual count of raptors is taken as they fly over Duluth during their migration south at Hawk Ridge - one of the premier birding sites in the US, located within the city limits! Recreation is available for any interests - from dragon boat races to speed skating, marathons, cross country skiing, mountain biking, hiking, downhill skiing on our own ski hill, sail boating, inline skating, rowing, and figure skating, snowshoeing, big lake fishing for salmon, inland lake fishing for walleye, stream fishing for trout, and don't forget hockey!! Visit www.duluth.com to see more about the city, and visit the SMDC website at www.smdc.org to see more about our institution. Apply online! Sheila Tapper HT(ASCP) Anatomic Pathology Supervisor SMDC Clinical Laboratory 407 East First Street Duluth, MN 55804 Telephone: 218-786-5472 Fax: 218-786-2369 This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any attachments. From AFoshey <@t> chw.org Fri Nov 9 16:50:07 2007 From: AFoshey <@t> chw.org (Foshey, Annette) Date: Fri Nov 9 16:50:16 2007 Subject: [Histonet] Glut 1 antibody Message-ID: <9E6D52F532809247BDA1783680E92C560B70294E@CHWEXC.chwi.chswi.org> Does anyone have a vendor for Glut 1 ? Thanks, Annette Foshey, HT (ASCP) Team Leader in Histology Children's Hospital of Wisconsin 414-266-6580 Fax 414-266-2779 afoshey@chw.org ************************** Children's Hospital and Health System recognizes that unencrypted e-mail is not secure and does not guarantee confidentiality. The confidentiality of replies to this message cannot be guaranteed unless the replies are encrypted. From MElliott <@t> mrl.ubc.ca Fri Nov 9 17:09:55 2007 From: MElliott <@t> mrl.ubc.ca (Mark Elliott) Date: Fri Nov 9 17:10:39 2007 Subject: [Histonet] New Hospital Wing Message-ID: <47347843.11C6.00D6.0@mrl.ubc.ca> Did the Respirologists blow a lot of "hot air" about the matter? *_NEW HOSPITAL WING_* When a panel of doctors was asked to vote on adding a new wing to their hospital, the Allergists voted to scratch it, and the Dermatologists advised not to make any rash moves. The Gastroenterologists had sort of a gut feeling about it, but the Neurologists thought the administration had a lot of nerve, and the Obstetricians felt they were all laboring under a misconception The Ophthalmologists considered the idea shortsighted; the Pathologists yelled, 'Over my dead body', while the Pediatricians said, 'Oh, grow up!' The Psychiatrists thought the whole idea was madness, the Radiologists could see right through it, and the Surgeons decided to wash their hands of the whole thing. The Internists thought it was a bitter pill to swallow, and the Plastic Surgeons said, 'This puts a whole new face on the matter.' The Podiatrists thought it was a step forward, but the Urologists felt the scheme wouldn't hold water. The Anesthesiologists thought the whole idea was a gas, and the Cardiologists didn't have the heart to say no. In the end, the Proctologists left the decision up to some a....... in administration. ***CONFIDENTIALITY NOTICE*** This electronic message is intended only for the use of the addressee and may contain information that is privileged and confidential. Any dissemination, distribution or copying of this communication by unauthorized individuals is strictly prohibited. If you have received this communication in error, please notify the sender immediately by reply e-mail and delete the original and all copies from your system. From mtarango <@t> nvcancer.org Fri Nov 9 17:16:49 2007 From: mtarango <@t> nvcancer.org (Tarango, Mark) Date: Fri Nov 9 17:17:28 2007 Subject: [Histonet] Glut 1 antibody In-Reply-To: <9E6D52F532809247BDA1783680E92C560B70294E@CHWEXC.chwi.chswi.org> Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F86@NVCIEXCH02.NVCI.org> I use GT15-M from Alpha Diagnostics (http://www.4adi.com/ 800-786-5777) with infantile hemangioma as the control. You should see staining restricted to the tumor endothelium. If you need control material, I'd be happy to send you some. Glad its Friday! Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Foshey, Annette Sent: Friday, November 09, 2007 2:50 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glut 1 antibody Does anyone have a vendor for Glut 1 ? Thanks, Annette Foshey, HT (ASCP) Team Leader in Histology Children's Hospital of Wisconsin 414-266-6580 Fax 414-266-2779 afoshey@chw.org ************************** Children's Hospital and Health System recognizes that unencrypted e-mail is not secure and does not guarantee confidentiality. The confidentiality of replies to this message cannot be guaranteed unless the replies are encrypted. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== From pruegg <@t> ihctech.net Sat Nov 10 10:07:32 2007 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Sat Nov 10 10:07:59 2007 Subject: [Histonet] frozen brain In-Reply-To: <254923.82101.qm@web50311.mail.re2.yahoo.com> References: <254923.82101.qm@web50311.mail.re2.yahoo.com> Message-ID: <001001c823b3$cde69ce0$0202a8c0@ihctechq9h2qof> After fixation I always infiltrate the tissue in 30% sucrose at least overnight before freezing. The morphology is very well preserved and frozen sections are great improved over trying to cut nbf fixed tissue frozen without sucrose infiltration. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of I-sanna Gibbons Sent: Thursday, November 08, 2007 1:01 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] frozen brain Hi all, Does anyone have a protocol for freezing fixed (10% formalin) brain tissue? Will cell morphology be compromised to a large extent? Thanks I-sanna __________________________________________________ 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 Sat Nov 10 10:10:13 2007 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Sat Nov 10 10:10:40 2007 Subject: [Histonet] Clone for Prog. Receptor In-Reply-To: <000101c82236$8754f750$d00f7ca5@lurie.northwestern.edu> References: <000101c82236$8754f750$d00f7ca5@lurie.northwestern.edu> Message-ID: <001101c823b4$2e2262b0$0202a8c0@ihctechq9h2qof> In my experience, the rab monoclonals for er and pr are just as sensitive or more so than the ms monoclonal, both using the same labeled polymer detection. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bernice Frederick Sent: Thursday, November 08, 2007 11:38 AM To: 'Drew Sally A.'; 'Histonet' Subject: RE: [Histonet] Clone for Prog. Receptor Per a lecture at the NSh mouse monocloncal antibodies have better affinity than rabbit monoclonals. We use 1A6 but then, we are following a clinical protocol and cannot deviate. Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Drew Sally A. Sent: Thursday, November 08, 2007 11:07 AM To: Histonet Subject: [Histonet] Clone for Prog. Receptor We currently are using the 1A6 clone for our progesterone receptor(PR) IHC. We were wondering how other people felt about rabbit monoclonal PR antibodies. Does anyone have strong preferences or experiences with other PR clones-mouse or rabbit-that they'd like to share? Thank you...! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 _______________________________________________ Histonet mailing list Histonet@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 Sat Nov 10 10:15:49 2007 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Sat Nov 10 10:16:15 2007 Subject: [Histonet] Clone for Prog. Receptor In-Reply-To: <473303A20200007700009027@gwmail4.harthosp.org> References: <473303A20200007700009027@gwmail4.harthosp.org> Message-ID: <001301c823b4$f60afda0$0202a8c0@ihctechq9h2qof> I do not have the clone, but I use rab monoclonal pr from Lab Vision with good results, I use a labeled polymer detection. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Thursday, November 08, 2007 10:40 AM To: Histonet; Drew Sally A. Subject: Re: [Histonet] Clone for Prog. Receptor We have used clone PgR636 (from Dako) for several years now with excellent results. 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 >>> "Drew Sally A." 11/08/07 12:07 PM >>> We currently are using the 1A6 clone for our progesterone receptor(PR) IHC. We were wondering how other people felt about rabbit monoclonal PR antibodies. Does anyone have strong preferences or experiences with other PR clones-mouse or rabbit-that they'd like to share? Thank you...! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 _______________________________________________ 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 koellingr <@t> comcast.net Sat Nov 10 10:40:49 2007 From: koellingr <@t> comcast.net (koellingr@comcast.net) Date: Sat Nov 10 10:41:01 2007 Subject: [Histonet] Clone for Prog. Receptor Message-ID: <111020071640.16383.4735DF110007C3EB00003FFF22007347489D09020704040A0105@comcast.net> Sally, You asked about PR but there is a lot of discussion about mouse monoclonals versus rabbit monoclonals in general and as there is a lot of "fact" floating around that is somewhere between questionable to outright fallacy, I thought I'd chime in. In a former life, we were involved with the company that originated the rabbit monoclonal Ab technology as well as some companies that they licensed their technology to. Anyone stating that rabbit monoclonals have higher affinities than their cousin mouse monoclonals as a general concept have some real experimental science to do. For reasons that would take a page or 2, I cannot do so here. Whether by IHC staining you get "more" staining or "less" staining signal with rabbits versus mouse Mab's, might have nothing what-so-ever to do with their relative affinities to tissue target. The readers digest version of this is that you need to do "Biacore" type (surface plasmon resonance technology) to determine affinities of antibodies. As it happens, one of the only (first?) articles published I've ever seen doing this to bring some sense to this controversial subject was published in the latest journal of the Journal of Histotechnology. Their results, I think will jolt a few of the preconceived notions that some have of the utter and absolute superiority of Rabbit Mab's over Mouse Mab's. A caveat, this was done by Vision Biosystems themselves. I think a great piece for a world-wide peer-reviewed journal would be to do their analysis (x5 or 6 antibodies) and with several more experiments and it would be to great service and utility for the entire IHC and general antibody world. I like Rabbit Ab's, have used and use them. Some utterly superior to Mouse Mab's for IHC in my hands. But some are not nearly as good and even terrible, in my hands and this may or may not have one thing to do with affinity. Raymond Koelling PhenoPath Laboratories Seattle, WA -------------- Original message -------------- From: > I do not have the clone, but I use rab monoclonal pr from Lab Vision with > good results, I use a labeled polymer detection. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard > Cartun > Sent: Thursday, November 08, 2007 10:40 AM > To: Histonet; Drew Sally A. > Subject: Re: [Histonet] Clone for Prog. Receptor > > We have used clone PgR636 (from Dako) for several years now with excellent > results. > > 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 > > >>> "Drew Sally A." 11/08/07 12:07 PM >>> > We currently are using the 1A6 clone for our progesterone receptor(PR) > IHC. > We were wondering how other people felt about rabbit monoclonal PR > antibodies. > Does anyone have strong preferences or experiences with other PR > clones-mouse or rabbit-that > they'd like to share? > Thank you...! > > Sally Ann Drew, MT(ASCP) > IHC/ISH Laboratory > University of Wisconsin Hosp. & Clinics > Madison, WI 53792 > (608)265-6596 > > _______________________________________________ > 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 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From middlebabs <@t> yahoo.com Sat Nov 10 11:13:26 2007 From: middlebabs <@t> yahoo.com (kathy machado) Date: Sat Nov 10 11:13:30 2007 Subject: [Histonet] Histology position open in Danville, VA Message-ID: <159926.95116.qm@web54307.mail.re2.yahoo.com> Hello all, We have a histochnecian position open at Danville Regional Medical Center in lovely Danville, Va. This is a small lab looking for a third full time histo tech. For more information email drhs.hr@lpnt.net or call 1-800-688-3762. Kathy Machado Danille, VA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From arnie.jimenez <@t> vel-lab.com Sat Nov 10 12:06:43 2007 From: arnie.jimenez <@t> vel-lab.com (arnie.jimenez@vel-lab.com) Date: Sat Nov 10 12:09:23 2007 Subject: [Histonet] Re: Away from Office Message-ID: <20071110180643.6022.qmail@plesk5.hostgator.com> Thank you for contacting Vel-Lab Research. We will be out of the lab starting Nov. 7th and will return on Nov. 12th. We will check e-mail regularly but will be unable to respond. Please do not mail any packages to our lab until Nov. 12th. Regards, Arnie Jimenez Vel-Lab Research From Eric.Hoy <@t> UTSouthwestern.edu Sat Nov 10 12:32:09 2007 From: Eric.Hoy <@t> UTSouthwestern.edu (Eric Hoy) Date: Sat Nov 10 12:32:15 2007 Subject: [Histonet] Re: Medical technology chat room In-Reply-To: <200711101800.lAAI0QlW027733@nlpi045.prodigy.net> Message-ID: Jonathan De La Rosa asked about a "Medical Technology" chat room. Try MEDLAB-L: http://www.ualberta.ca/~pletendr/MEDLAB-L.html It's a nice discussion group, and Pat Letendre, the listowner, has done an excellent job. Eric Hoy =================================================== Eric S. Hoy, Ph.D., SI(ASCP) Clinical Associate Professor Department of Medical Laboratory Sciences The University of Texas Southwestern Medical Center Dallas, Texas Email: Eric.Hoy@UTSouthwestern.edu =================================================== From mshaeffer <@t> cox.net Sat Nov 10 16:54:00 2007 From: mshaeffer <@t> cox.net (Marc Shaeffer) Date: Sat Nov 10 16:54:11 2007 Subject: [Histonet] AO "Spencer" 820 Locking Lever Message-ID: <006701c823ec$959ede90$6500a8c0@youro0kwkw9jwc> Does anyone know where I can find a =93locking lever=94 for an AO = =93Spencer=94 820 microtome? We have three of these instruments in a research lab and the locking lever is broken on all three. Thanks in advance, =20 Marc Shaeffer Histotechnology Student No virus found in this outgoing message. Checked by AVG Free Edition.=20 Version: 7.5.503 / Virus Database: 269.15.27/1121 - Release Date: = 11/9/2007 7:29 PM =20 From gayle.callis <@t> bresnan.net Sun Nov 11 11:35:57 2007 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Sun Nov 11 11:35:14 2007 Subject: [Histonet] Gayle Callis is retired, however Message-ID: <000c01c82489$51a97a40$6501a8c0@DHXTS541> Dear Histonetters I decided to retire this past September from routine daily work. However, I will be consulting with one researcher who needs immunostaining and cryotomy skills but not on a full time basis. He threatened to chain me to his cryostat but more than that, it will keep me active both mentally and in the lab. Retirement does not mean we lose our skills. I will remain active with the National Society for Histotechnology and the Journal of Histotechnology -in particular, The Writing Partners Program to help individuals less skilled in scientific manuscript preparation but have so much to share through their methods, techniques, hints, modifications, and so on. NSH and their sponsors have been an important part of my career by giving so much in recognition but also the opportunity to study abroad. It has been a long career in this profession, starting in 1962 with many wonderful changes along the way - computers with Internet access to anything we need, better instrumentation with automation, fabulous microsocopy advances, superb cryostats, plastic methods for light and electron microscopy, undecalcified bone techniques with MMA, and more than anything the entry of immunohistochemical staining and molecular biology techniques into our field. The latter two could be compared to the universe's "Big Bang Theory" - at a rate of expansion to keep us on our histo-toes forever. One thing to be noted are the routine staining methods we use today, including decalcification. These much the same as when they were first published long before I entered histotechnology. I hope those new to the profession will become students of histotechnology and read publications about the tried and true classic methods. Any many of us still perform our duties as they were done early on - doing things by hand i.e. processing. It has been a delightful experience to be part of the Histonet participation which has broadened my histotechnology experience so vitally needed to stay current in this profession. The discussions were informative and very useful, but most importantly are the wonderful people i have met through this list. These individuals are now some of my closest friends and colleagues who enrich my life daily, both personally and histotechnologically. The Histonet format has provided me (and will contine to do so) with so much - the excellent discussions, a means of finding information/products, the right to disagree or reply privately, but most of all - meeting people on an international basis. I will subscribe from time to time to participate on Histonet, but may be using a different email address at home, e.g. a business address. My university email address is no longer in service so if anyone has tried to contact me, the email was probably rejected, returned or ITC deleted it. Forwarding was not an option since due to spam issues. My contact information is on the NSH website, members only webpage, and some non NSH members also have my current address. For those new to Histonet, don'r forget to use the Histonet Archives - it is a vast resource of unbelievable hints and help on improving one's histotechnics. Take care, stay in touch, see you on Histonet Gayle M. Callis MT,HT,HTL(ASCP) Bozeman MT From gayle.callis <@t> bresnan.net Sun Nov 11 12:00:26 2007 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Sun Nov 11 11:59:42 2007 Subject: [Histonet] Rat brain in 70% alcohol for cryotomy Message-ID: <000801c8248c$bd386430$6501a8c0@DHXTS541> Amy, You wrote We received a few whole rat brains in 70% EtOH after perfused with 10% BF. Now they changed mind want go for frozen. The want Cresyl Violet stain. Is it doable? How The problem is that 70% alcohol acts as antifreeze, and unless you remove the alcohol, the brain will not freeze properly. The alcohol can be replaced with a sucrose cryoprotection, needed even after NBF fixaiton for proper snap freezing. You could sucrose cryoprotect the brains in order to prepare them for snap freezing and cryotomy, but it may take several days using an increasing concentration/sucrose gradient to replace the alcohol with sucrose solution . You can start in 15% sucrose, 20% sucrose and then 30% sucrose. I am not sure of timing for a whole rat brain, but overnight in each change at 4C may do the job. It may be easier to just process into paraffin and do the cresyl echt violet on paraffin sections instead, unless they are asking for thick sections? We do cresyl violet staining on paraffin sections instead of frozen sections, but the stain can be done on frozen sections. Histonet Archives has discussed sucrose gradients, the problem you presented and also doing cresyl echt violet on frozen sections - you may want to do a search to find out what others have discussed. Good luck, Gayle Callis MT,HT,HTL(ASCP) From arnie.jimenez <@t> vel-lab.com Sun Nov 11 12:05:56 2007 From: arnie.jimenez <@t> vel-lab.com (arnie.jimenez@vel-lab.com) Date: Sun Nov 11 12:08:48 2007 Subject: [Histonet] Re: Away from Office Message-ID: <20071111180556.25802.qmail@plesk5.hostgator.com> Thank you for contacting Vel-Lab Research. We will be out of the lab starting Nov. 7th and will return on Nov. 12th. We will check e-mail regularly but will be unable to respond. Please do not mail any packages to our lab until Nov. 12th. Regards, Arnie Jimenez Vel-Lab Research From gayle.callis <@t> bresnan.net Sun Nov 11 12:42:26 2007 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Sun Nov 11 12:41:40 2007 Subject: [Histonet] CD4 and FOXP3 dual staining using same species primary hosts Message-ID: <000c01c82492$9b3ab440$6501a8c0@DHXTS541> Ann, We do two rat antiMouse primaries for immunofluorescent staining routinely, and have done triple staining also, without problems. You must do sequential staining with the primaries. Our favorite method is to use one biotinylated antibody, and that one is used after the first primary/secondary application. For immunofluorescent work we do NOT use BSA or any protein carriers in our buffers, but do add 0.025% Tween 20 for sheeting action. Rinsing well is critical for IFA work. Spin all fluorophores after dilution and just before application to get rid of any fluorophore labelled protein aggregates (glowing garbage!). Our frozen sections are air dried overnight at RT, and fixed the next day - if you use acetone, 10 min at 4C, air dry then proceed to buffer after getting rid of acetone fumes. Normal serum block 5% donkey (or goat depending on host of secondary) serum 30 min RT streptavidin/biotin block is needed. (Vector kit) 1. FOXP3 primary rat antimouse monoclonal diluted in 5% donkey serum, this may not need overnight incubation unless you have determined you need overnight for this primary. 2. Donkey antiRat F(ab')2 frag of IgG - Rhodamine Red X, adsorbed to mouse tissue 1:300 or so diluted in 5% donkey serum. 30 min RT (Jackson Immunoresearch for this antibody) For secondary antibody, you can purchase a different one from Molecular Probes as you did, and the Alexa 594 it will have better spectral separation from the 488. The seconday should be adsored to the mouse. If the secondary is made in goat then use goat serum instead. 3. Rat serum, 5% for 30 minutes in order to ensure the secondary antibody is bound to rat, and does NOT recognize the next rat antimouse primary, even if it is biotinylated. Rinse well. 4. CD4-biotinylated (BD Biosciences) 1:500 in 5% donkey serum/1.25% mouse serum 30 minutes at RT 5. Strepavidin-488 diluted in buffer with Tween, NO SERUM with this step. Molecular Probes gives a dilution range up to 1:1000, and this will work, incubate in dark for 30 min, RT. Rinse well with pure buffer and mount coverslip with Prolong Gold antifade reagent, ready to use from Molecular Probes The denaturing solution is NOT needed when doing this method. Using a biotinylated primary is a good way to avoid cross reaction to a secondary antibody previously used, but the rat serum block is critical. If you want to see a photograph of this type of staining method, I will be happy to attach on to you privately, but will need your email address. Gayle Callis MT,HT,HTL(ASCP) ******************************************************************** Hi Ann, I think the problem is that you are blocking with rat serum. You wouldn't block with rat serum before your first primary, and you shouldn't for your second primary since both are rat antibodies. There are several ways you could try to accomplish dual staining of 2 rat antibodies. I think the easiest, since you are doing fluorescent on frozens, is use BD's rat x mouse CD4 FITC (green cytoplasm). It works very well. Ebioscience has AF conjugated primaries, so you should be able to get rat x FOXP3 AF555 or AF594 (red nucleus). Then you could just cocktail them and do single step staining. Or you could try Probes (Invitrogen's) Zenon labeling kits, and directly conjugate your rat antibodies yourself. (I don't know if they have kits for rat primaries though) Thirdly, you could try following your protocol for the first primary, adding Biocare Medicals' Denaturing solution to seal the antigen site (since the first is cytoplasmic) then the second would have access to the nuclear site. 1. 5% normal goat serum block 20 min 2. 1* antibody CD4 1:750 in diluent O/N 4C 3. Seconday AF594 GT anti-rat 1:1k in 1xTBS 1h RT 4. Biocare Medicals' Denaturing solution (follow manuf instructions) 5. Serum block: 5% normal DONKEY serum 30 min 6. 1* antibody FOXP3 1:100 in diluent 1h RT7. Secondary AF488 Donkey anti-rat in TBS 1h RT From mtarango <@t> nvcancer.org Sun Nov 11 17:24:59 2007 From: mtarango <@t> nvcancer.org (Tarango, Mark) Date: Sun Nov 11 17:25:20 2007 Subject: [Histonet] QIHC Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F92@NVCIEXCH02.NVCI.org> To anyone who has taken the QIHC exam, what kinds of questions does it ask? I've got a few books from the reading list, but I have no idea where to focus my attention. It is more about staining techniques or immunophenotyping? Quality control stuff? What's on the test that you might not expect? Is it a difficult exam? Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== From Kemlo.Rogerson <@t> waht.swest.nhs.uk Mon Nov 12 02:25:35 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Nov 12 02:25:47 2007 Subject: [Histonet] Fwd: Artefacts Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F14A@wahtntex2.waht.swest.nhs.uk> He's correct the 'pictures' aren't there. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; Nonviolence means avoiding not only external physical violence but also internal violence of spirit. You not only refuse to shoot a man, but you refuse to hate him. --Martin Luther King Jr. This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From octavio109 <@t> hotmail.com Mon Nov 12 07:47:58 2007 From: octavio109 <@t> hotmail.com (Corinthia D. Emanuel) Date: Mon Nov 12 07:48:05 2007 Subject: [Histonet] FW: Atlanta, GA derm path lab seeks techs & assistants In-Reply-To: References: Message-ID: Some previous experience highly preferred for immediate part time positions early morning, late afternoon, evening or weekends. ASAP, send email of interest with availability and paste resume within the email, please. Include at least two (direct supervisor) references, please. Address email to cemanuel@myfamilyderm.com ________________________________> Subject: Atlanta, GA derm path lab seeks techs & assistants> Date: Mon, 12 Nov 2007 08:42:03 -0500> From: cemanuel@myfamilyderm.com> To: histonet@lists.utsouthwestern.edu> CC: octavio109@hotmail.com>> Some previous experience highly preferred for part time positions early morning, late afternoon, evening or weekends.> ASAP, send email of interest with availability and paste resume within the email, please.> Include at least two (direct supervisor) references, please.> Address email to cemanuel@myfamilyderm.com _________________________________________________________________ Boo!?Scare away worms, viruses and so much more! Try Windows Live OneCare! http://onecare.live.com/standard/en-us/purchase/trial.aspx?s_cid=wl_hotmailnews From tjey <@t> hotmail.com Mon Nov 12 08:40:38 2007 From: tjey <@t> hotmail.com (Tanya Ewing-Finchem) Date: Mon Nov 12 08:40:45 2007 Subject: [Histonet] Histology Job In Tucson In-Reply-To: References: Message-ID: Histotechnician III IRC4399 To support the functions of the Histology Department by performing the following duties. PRIMARY RESPONSIBILITIES AND ACCOUNTABILITY:The Duties of a Histotechnician III involve: Grossing, processing, embedding tissue sample. Maintaining lab records and equipment. Qualifying tissue blocks. Section tissue samples onto slides. Perform routine H&E stains. Perform Verification/Validation Studies. (Follow protocols, communicate with R &D, trouble-shoot). Grading instrument manufacturing test slides. Perform Oracle transactions concerning manufactured control slides kits. QC testing of manufactured control slides kits. Monitors work to ensure quality, and continuously promotes Quality First Time. Qualifications:To perform this job successfully, an individual must be able to perform each essential duty satisfactorily. The requirements listed below are representative of the knowledge, skill, and/or ability required. Reasonable accommodations may be made to enable individuals with disabilities to perform the essential functions.Education/Experience: Minimum of a Associates Degree in a related field and 3-5 years experience in the field of histology. HT (ASCP) Certified. Q-IHC (ASCP) certified or High School Diploma and 5-7 years experience HT(ASCP) Certified. Required Training: Continuing education in the field of histology, immunohistochemistry and insitu. Updates of FDA/ health regulations in dealing with handling human tissue and chemical solvents. Ventana offers a complete benefits package. Employees can select options that meet their individual needs incl medical, dental, vision; company paid life insurance, 401k and Employee Stock Purchase Plan. To be considered for the position you must register at https://careers.ventanamed.com. Ventana Medical Systems, Inc. 1910 Innovation Park Drive Tucson, AZ 85755 Ventana is an equal opportunity employer. M/F/D/V From thisisann <@t> aol.com Mon Nov 12 08:48:51 2007 From: thisisann <@t> aol.com (thisisann@aol.com) Date: Mon Nov 12 08:49:15 2007 Subject: [Histonet] Gold colored slides Message-ID: <8C9F35246600E06-CE8-86B@FWM-D19.sysops.aol.com> Can someone please let me know where I can order gold colored slides, at a reasonable price!! Thank you, Ann Angelo ________________________________________________________________________ Email and AIM finally together. You've gotta check out free AOL Mail! - http://mail.aol.com From emilietrudelle <@t> hotmail.com Mon Nov 12 09:07:41 2007 From: emilietrudelle <@t> hotmail.com (emilie trudelle) Date: Mon Nov 12 09:07:48 2007 Subject: [Histonet] unsuscribe Message-ID: Please unsuscribe me. thanks _________________________________________________________________ Envoie un sourire, fais rire, amuse-toi! Employez-le maintenant! http://www.emoticonesgratuites.ca/?icid=EMFRCA120 From trinimaican2501 <@t> yahoo.com Mon Nov 12 09:32:54 2007 From: trinimaican2501 <@t> yahoo.com (I-sanna Gibbons) Date: Mon Nov 12 09:33:02 2007 Subject: [Histonet] frozen bat brain Message-ID: <479694.71699.qm@web50306.mail.re2.yahoo.com> Thanks for all the suggestions! Initially I was producing paraffin sections of bat brains but the results were not satisfactory. I am recording cell structure of the nuclei in the brainstem. I still have a number of brains fixing in 10% formalin and now want to change to produce frozen sections without compromising cell structure. Most of you suggested starting off by immersing the brain in 30% sucrose (cryoprotectant) until the tissue sinks. Does anyone have a detailed procedure (material, duration, temperature) for whole brains that can follow after this step or know of a site to which I can be directed that will provide me with that information? Thanks a lot I-sanna __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Heather.D.Renko <@t> osfhealthcare.org Mon Nov 12 09:37:50 2007 From: Heather.D.Renko <@t> osfhealthcare.org (Renko, Heather D.) Date: Mon Nov 12 09:38:12 2007 Subject: [Histonet] RE: large specimen cut off times Message-ID: <40026EDDE64CDA47AB382C52619ACD3C06612D5C@pmc-rfd-mx01.intranet.osfnet.org> Hello fellow histo netters, If any of you have a specific policy for cut off times for specimens (i.e. large mainly, colon etc.)in your department, can you please respond to me personally. I am developing a policy and would like to know what the standards in the group are-I have my pre conceived thoughts of what should be the times but, I would really like to get an idea of what other facilities do. My processor begins its cycle at 3:30pm each day with three changes of formalin and goes into the first alcohol(70%) at 7pm each night. Thank you in advance. Heather D. Renko, Histology Coordinator OSF Saint Anthony Medical Center 5666 East State Street Rockford, Illinois 61108 Direct: 815-395-5410 Heather.D.Renko@osfhealthcare.org ============================================================================== The information in this message is confidential and may be legally privileged. Access to this message by anyone other than the addressee is not authorized. If you are not the intended recipient, or an agent of the intended recipient, any disclosure, copying, or distribution of the message or any action or omission taken by you in reliance on it, is prohibited and may be unlawful. If you have received this message in error, please contact the sender immediately and permanently delete the original e-mail, attachment(s), and any copies. ============================================================================== From jengirl1014 <@t> yahoo.com Mon Nov 12 12:14:18 2007 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Mon Nov 12 12:14:21 2007 Subject: [Histonet] Getting a Tissue Processor Message-ID: <750683.40287.qm@web62408.mail.re1.yahoo.com> My lab is FINALLY getting a Shandon Excelsior Tissue Processor. I was wondering if anyone had any good protocols for the machine that would be great for mouse tissue...mainly the GI trac and liver and kidneys. Thanks for all of your help! Jen __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From vvancleave <@t> hendrickhealth.org Mon Nov 12 13:54:54 2007 From: vvancleave <@t> hendrickhealth.org (Vancleave, Vince) Date: Mon Nov 12 13:55:29 2007 Subject: [Histonet] non pathologist grossing vs processing continued References: <740F77F6594C20458BDC2A4A500D8D4150163F@EMAIL.hhs.hendrickhealth.org> Message-ID: <740F77F6594C20458BDC2A4A500D8D41501642@EMAIL.hhs.hendrickhealth.org> From: Vancleave, Vince Sent: Wed 11/7/2007 2:39 PM To: histonet@lists.utsouthwestern.edu Cc: Webster, John A. M.D. Subject: Still not sure about this low complexity or not "The thing you have to remember, Vince, is that CLIA and CAP are two different, completely separate entities. CLIA is government controlled and CAP is private. Whereas CAP recently decided to separate grossing into two different complexities, CLIA has not. Bottom line...if you are CLIA licensed you MUST follow the CLIA standard. If not, then you can do whatever you like. Charles Embrey Jr., PA(ASCP)" Charles makes a great point but: I'm just not sure I'm sold on this yet, b/c also in the CLIA regs it states in sec. 493.559 about exemption from CLIA, via using a private accreditation agency (which is what CAP is), and states clearly that this agency can only be deemed acceptable if their requirements for the laboratory are equivalent or more stringent and the laboratory would meet CLIA requirements if the laboratory had not been granted deemed status. Ya'll are right, this is a mess! So, wouldn't this essentially mean that according to CLIA, since CAP is an acceptable private agency, they are giving the ok for CAP's decision on this? Wouldn't CAP be unallowed to do what they do if it was less stringent than CLIA? I also find nothing in the entire CLIA 88' that indicates that this would not be ok. In fact, CLIA has a criteria for categorization of complexity: http://www.fda.gov/cdrh/clia/categorization.html . And in their database of tests that they have said has to absolutely be a certain way, I find no reference to tissue description, gross examination, tissue examination, or processing. Have I stumped anyone yet, or is there some more things I am missing? I really appreciate all ya'lls willingness to re-visit this issue again. I guess I am a stuborn monkey! Thanks, Vince Van Cleave, BS, PA(ASCP), HTL, HT Laboratory Manager and Pathologist Assistant (325)670-6528 Clinical Pathology Associates Abilene, TX PRIVILEGED AND CONFIDENTIAL NOTICE: The information contained in this e-mail may be confidential and/or privileged. This e-mail is intended to be reviewed initially by only the individual named above. If the reader of this e-mail is not the intended recipient or a representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail, or the information contained herein, is prohibited. If you have received this e-mail in error, notify the above sender, send the message back and then delete it. Thank you for your assistance. Hendrick Health System 1900 Pine St. Abilene, Texas 79601 - 325.670.2000 79601 - 325.670.2000 From Charles.Embrey <@t> carle.com Mon Nov 12 14:28:36 2007 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Mon Nov 12 14:29:57 2007 Subject: [Histonet] non pathologist grossing vs processing continued In-Reply-To: <740F77F6594C20458BDC2A4A500D8D41501642@EMAIL.hhs.hendrickhealth.org> Message-ID: <44780C571F28624DBB446DE55C4D733A1FE56A@EXCHANGEBE1.carle.com> Hi Vince, Sorry to take so long in getting back to you but things here have been very busy. By CLIA's guidelines it sounds as though CAP has removed themselves as an "acceptable" private accreditation when they loosened the restrictions on grossing. CLIA's database may not list grossing but CLIA did publish a guideline to clarify their position: This is the CLIA '88 guideline for high complexity testing personnel- CLIA '88 493.1489 "On or before April 24 1995 (I) be a high school graduate or equivalent; and (b) have documentation of training appropriate for the test performed before analyzing patient specimens"...After that date it requires an associate degree in a biological or chemical science or medical laboratory technology -or- qualify as a medical technologist with a bachelor's degree from an accredited institution -or- earned a bachelor's degree in a chemical, physical, biologic or clinical laboratory science. A few years ago CLIA put out this interpretive guideline, (printed below), to clear up the question of grossing. As you see, it references 493.1498 (the above "high complexity guideline"). They basically sum up that if you examine/describe the tissue for record noting color weight or measurement then it is grossing and falls under 493.1498. Tissue processing as CAP calls it involves noting size and other characteristics and qualifies as grossing under CLIA. The new federal register CLIA interpretive guidelines appendix C subpart M effective April 24, 2003 states in section 493.1461(e) "In the case if gross examinations, the technical supervisor may delegate to individuals qualified under 493.1498 the responsibility for the physical examination/description, including color, weight, measurement, and other characteristics of the tissue; or other mechanical procedures for which a specific written protocol has been developed." I hope this helps. I don't know which side will budge first but I have heard whisperings of CLIA doing something in the near future but what ever it is it will probably only muddy the water further. All I can say is that CLIA is what happens when a government bureaucracy gets involved with healthcare. Charles Embrey PA(ASCP) Histology Manager Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vancleave, Vince Sent: Monday, November 12, 2007 1:55 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] non pathologist grossing vs processing continued From: Vancleave, Vince Sent: Wed 11/7/2007 2:39 PM To: histonet@lists.utsouthwestern.edu Cc: Webster, John A. M.D. Subject: Still not sure about this low complexity or not "The thing you have to remember, Vince, is that CLIA and CAP are two different, completely separate entities. CLIA is government controlled and CAP is private. Whereas CAP recently decided to separate grossing into two different complexities, CLIA has not. Bottom line...if you are CLIA licensed you MUST follow the CLIA standard. If not, then you can do whatever you like. Charles Embrey Jr., PA(ASCP)" Charles makes a great point but: I'm just not sure I'm sold on this yet, b/c also in the CLIA regs it states in sec. 493.559 about exemption from CLIA, via using a private accreditation agency (which is what CAP is), and states clearly that this agency can only be deemed acceptable if their requirements for the laboratory are equivalent or more stringent and the laboratory would meet CLIA requirements if the laboratory had not been granted deemed status. Ya'll are right, this is a mess! So, wouldn't this essentially mean that according to CLIA, since CAP is an acceptable private agency, they are giving the ok for CAP's decision on this? Wouldn't CAP be unallowed to do what they do if it was less stringent than CLIA? I also find nothing in the entire CLIA 88' that indicates that this would not be ok. In fact, CLIA has a criteria for categorization of complexity: http://www.fda.gov/cdrh/clia/categorization.html . And in their database of tests that they have said has to absolutely be a certain way, I find no reference to tissue description, gross examination, tissue examination, or processing. Have I stumped anyone yet, or is there some more things I am missing? I really appreciate all ya'lls willingness to re-visit this issue again. I guess I am a stuborn monkey! Thanks, Vince Van Cleave, BS, PA(ASCP), HTL, HT Laboratory Manager and Pathologist Assistant (325)670-6528 Clinical Pathology Associates Abilene, TX PRIVILEGED AND CONFIDENTIAL NOTICE: The information contained in this e-mail may be confidential and/or privileged. This e-mail is intended to be reviewed initially by only the individual named above. If the reader of this e-mail is not the intended recipient or a representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail, or the information contained herein, is prohibited. If you have received this e-mail in error, notify the above sender, send the message back and then delete it. Thank you for your assistance. Hendrick Health System 1900 Pine St. Abilene, Texas 79601 - 325.670.2000 79601 - 325.670.2000 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SHargrove <@t> urhcs.org Mon Nov 12 16:52:19 2007 From: SHargrove <@t> urhcs.org (SHargrove@urhcs.org) Date: Mon Nov 12 16:52:46 2007 Subject: [Histonet] Susie Hargrove is out of the office. Message-ID: I will be out of the office starting 11/12/2007 and will not return until 11/19/2007. I will respond to your message when I return. From MichelM9 <@t> chw.edu.au Mon Nov 12 17:27:51 2007 From: MichelM9 <@t> chw.edu.au (Michelle McDonald) Date: Mon Nov 12 17:29:15 2007 Subject: [Histonet] deplasticizing PMMA sections for staining Message-ID: <47BD6E7614A693499453835D47E36F7003882B4F@hedwig.nch.kids> We actually use Acetone to deplasticise MMA sections. Only takes 2 changes for 10 mins each. Michelle Dr Michelle McDonald B.MedSci, PhD Research Officer Orthopaedic Research Dept. The Children's Hospital Westmead. Westmead NSW 2145 Australia Ph. +612 9845 1451 Fax. +612 9845 3078 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Tuesday, 6 November 2007 6:22 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] deplasticizing PMMA sections for staining In my experience, xylene is a rather poor solvent for PMMA. It works much faster at 60 degrees, but don't try heating flammable solvents in your oven unless you are certain it is certified explosion-proof (most paraffin ovens are not). Even then, unless you have the oven in or at least next to a fume hood, you'll get a lot of xylene vapors in the air as a result of heating it. I deplasticise PMMA with 2-methoxyethyl acetate, which only takes a couple of hours at room temperature. This has a rather strong though not unpleasant odor, and should be used in the hood. _______________________________________________ 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 note 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 san_tin <@t> biolab.com.sg Mon Nov 12 19:34:47 2007 From: san_tin <@t> biolab.com.sg (San Tin) Date: Mon Nov 12 19:34:40 2007 Subject: [Histonet] MyoD1 Message-ID: Dear All, Can anyone give me the protocol of MyoD1 clone 5.8A on Ventana IHC autostainer? I have problem with staining. Thanks San From cfarish <@t> csu.edu.au Mon Nov 12 21:03:54 2007 From: cfarish <@t> csu.edu.au (Farish, Craig) Date: Mon Nov 12 21:04:05 2007 Subject: [Histonet] RE: TGIF Message-ID: <79A000B60AFE8045BA2581119DFEC44402A251D4@ESWW01.CSUMain.csu.edu.au> And in return: A surgeon, pathologist, physician, psychiatrist and radiologist go duck hunting out on a pond. As the first duck flies over head, the psychiatrist takes aim, ....but doesn't fire. "What the hell happened?" demands the surgeon (they're known for their retiring manner) "Well, says the psychiatrist, I know its a duck, and YOU know its a duck - but does the duck know its a duck?" The second bird flies over head, and the physician takes aim.....but doesn't fire. "What now???" roars the surgeon. Well, says the physician, in a recent article I read in the New England Journal of Medicine, 12% of ducks are actually Terns, and they're a protected species.... The third duck flies over, and the radiologist takes aim......then paddles the boat further into the pond, then takes aim, then paddles back a bit, then takes a further aim....and by this time the duck has gone. "Jesus wept!" explodes the surgeon, what the f*** were you doing? Well, says the radiologist - every time I took aim, I realised that I needed another view. After lunch, they re-group for more hunting. This time the surgeon gets out his double barrelled, gold plated bazooka, blows his duck whistle - and as a flock of birds swarm over head, he fires randomly and enthusiastically into the air. Objects rain out of the sky. When he had finally finished blasting away to the heavens - he turned to the pathologist and said: "row over to all those bodies, and tell me if any of them was a duck" Craig (Joe) Farish Technical Officer School of Agricultural and Veterinary Sciences Charles Sturt University Wagga Wagga NSW Australia ' I don't want to achieve immortality through my work, I want to achieve immortality through not dying' - Woody Allen From ombadda3 <@t> gmail.com Tue Nov 13 04:54:35 2007 From: ombadda3 <@t> gmail.com (K M) Date: Tue Nov 13 04:54:47 2007 Subject: [Histonet] Claudin-1 as a prognostic factor in colorectal cancer Message-ID: Hi all I am here again and I start do my Msc and the topic I have chosen is: Claudin-1 as a prognostic factor in colorectal cancer. I will tell you each step I will do so I can benefit from your valuable experiences and knowledge. Thanks again to all members of this valuable forum -- Khalaf Mobile Phone: 0020127920404 Egypt From DixonM <@t> vetmed.ufl.edu Tue Nov 13 07:18:06 2007 From: DixonM <@t> vetmed.ufl.edu (MaryAnn Dixon) Date: Tue Nov 13 07:18:26 2007 Subject: [Histonet] staining of negative controls with IHC Message-ID: <47395DBE.6EA2.00FD.0@vetmed.ufl.edu> We are currently retrieving animal tissue in a pressure cooker with a program of 125 degrees for 30 secs. This protocol was taken from biocare but I'm sure this is for human tissue. Most of the tissue is retrieved with Biocare's Reveal (pH 6.0). We are receiving some background staining of our negative controls whereas if we did not retrieve them, there is none. We were staining for KI-67 which is a nuclear stain. I am using a tonsil for the control. The negative control showed some sporatic cytoplasmic staining on the Universal negative control from biocare as well as deionized water. Since then we have taken the temperature down to 115 degrees and increased the time to 25 minutes. It seems to have decreased the cytoplasmic staining but there is still some lingering. Anyone out there have any ideas??? I'm sure there is a better protocol for retrieving animal tissues. It has to be a time and temperature thing! MaryAnn Dixon Biological Scientist Anatomic Pathology University of Florida School of Veterinary Medicine 352-392-2235 ext 4517 From rjbuesa <@t> yahoo.com Tue Nov 13 07:32:35 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 13 07:32:40 2007 Subject: [Histonet] staining of negative controls with IHC In-Reply-To: <47395DBE.6EA2.00FD.0@vetmed.ufl.edu> Message-ID: <428387.48113.qm@web61222.mail.yahoo.com> There are many who prefer the pressure cooker because of the low incubation period but I personally did not like it because of background issues as you describe and because some tissues peel-off the slides (too much heat). Because of that I ended using a HIER step in a buffer (citrate or EDTA depending on the pH) initially heated to boliling temperature in a microwave oven and the actual HIER to take place in a steamer during 20 minutes + 20 minutes out of the steamer. All problems were resolved this way. Give it a try. Ren? J. MaryAnn Dixon wrote: We are currently retrieving animal tissue in a pressure cooker with a program of 125 degrees for 30 secs. This protocol was taken from biocare but I'm sure this is for human tissue. Most of the tissue is retrieved with Biocare's Reveal (pH 6.0). We are receiving some background staining of our negative controls whereas if we did not retrieve them, there is none. We were staining for KI-67 which is a nuclear stain. I am using a tonsil for the control. The negative control showed some sporatic cytoplasmic staining on the Universal negative control from biocare as well as deionized water. Since then we have taken the temperature down to 115 degrees and increased the time to 25 minutes. It seems to have decreased the cytoplasmic staining but there is still some lingering. Anyone out there have any ideas??? I'm sure there is a better protocol for retrieving animal tissues. It has to be a time and temperature thing! MaryAnn Dixon Biological Scientist Anatomic Pathology University of Florida School of Veterinary Medicine 352-392-2235 ext 4517 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Never miss a thing. Make Yahoo your homepage. From AGrobe2555 <@t> aol.com Tue Nov 13 09:39:46 2007 From: AGrobe2555 <@t> aol.com (AGrobe2555@aol.com) Date: Tue Nov 13 09:39:56 2007 Subject: [Histonet] staining of negative controls with IHC Message-ID: You don't say what detection system you are using....However, if you are using a Biotin-avidin/streptavidin detection, the background may be due to the exposure of endogenous biotin by HIER. You may need to incorporate an appropriate blocking agent/system Albert Albert C. Grobe, PhD International Heart Institute of Montana Foundation Tissue Engineering Lab, Saint Patrick Hospital ************************************** See what's new at http://www.aol.com From mtarango <@t> nvcancer.org Tue Nov 13 10:24:00 2007 From: mtarango <@t> nvcancer.org (Tarango, Mark) Date: Tue Nov 13 10:24:21 2007 Subject: [Histonet] staining of negative controls with IHC In-Reply-To: <47395DBE.6EA2.00FD.0@vetmed.ufl.edu> Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F9A@NVCIEXCH02.NVCI.org> You probably don't even need to get up to boiling for ki-67. Try 30 minutes at 95 C in your pressure cooker. Even on a "negative" control you're bound to see something stain positive for ki-67, since it's a proliferation marker... although it shouldn't be cytoplasmic. I'm just guessing there's got to be a cell or two in the tissue that's dividing. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of MaryAnn Dixon Sent: Tuesday, November 13, 2007 5:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] staining of negative controls with IHC We are currently retrieving animal tissue in a pressure cooker with a program of 125 degrees for 30 secs. This protocol was taken from biocare but I'm sure this is for human tissue. Most of the tissue is retrieved with Biocare's Reveal (pH 6.0). We are receiving some background staining of our negative controls whereas if we did not retrieve them, there is none. We were staining for KI-67 which is a nuclear stain. I am using a tonsil for the control. The negative control showed some sporatic cytoplasmic staining on the Universal negative control from biocare as well as deionized water. Since then we have taken the temperature down to 115 degrees and increased the time to 25 minutes. It seems to have decreased the cytoplasmic staining but there is still some lingering. Anyone out there have any ideas??? I'm sure there is a better protocol for retrieving animal tissues. It has to be a time and temperature thing! MaryAnn Dixon Biological Scientist Anatomic Pathology University of Florida School of Veterinary Medicine 352-392-2235 ext 4517 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== From lesley.bechtold <@t> jax.org Tue Nov 13 10:01:02 2007 From: lesley.bechtold <@t> jax.org (Lesley Bechtold) Date: Tue Nov 13 10:32:14 2007 Subject: [Histonet] automated paraffin embedding systems Message-ID: <20071113110102345.00000002152@spikey> Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 From rjbuesa <@t> yahoo.com Tue Nov 13 11:03:48 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 13 11:03:53 2007 Subject: [Histonet] automated paraffin embedding systems In-Reply-To: <20071113110102345.00000002152@spikey> Message-ID: <284972.24114.qm@web61211.mail.yahoo.com> This embedding instrument is manufactured by Sakura. It is not that the instrument orients the tissue, that is done when the tissues are placed in the cassette and held in place by a plastic frame that will be sectioned along with the tissue when the block is prepared. To function ideally it requires a special type of cassette and cassette holder (that is used alsao by the Sakura Xpress microwave tissue processor). Ren? J. Lesley Bechtold wrote: Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From mpence <@t> grhs.net Tue Nov 13 11:09:57 2007 From: mpence <@t> grhs.net (Mike Pence) Date: Tue Nov 13 11:10:07 2007 Subject: [Histonet] automated paraffin embedding systems In-Reply-To: <284972.24114.qm@web61211.mail.yahoo.com> Message-ID: <661949901A768E4F9CC16D8AF8F2838CA1C7A8@IS-E2K3.grhs.net> Is anyone out there using this function of the Sakura Xpress? Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, November 13, 2007 11:04 AM To: Lesley Bechtold; Histology Network Subject: Re: [Histonet] automated paraffin embedding systems This embedding instrument is manufactured by Sakura. It is not that the instrument orients the tissue, that is done when the tissues are placed in the cassette and held in place by a plastic frame that will be sectioned along with the tissue when the block is prepared. To function ideally it requires a special type of cassette and cassette holder (that is used alsao by the Sakura Xpress microwave tissue processor). Ren? J. Lesley Bechtold wrote: Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lesley.bechtold <@t> jax.org Tue Nov 13 11:26:32 2007 From: lesley.bechtold <@t> jax.org (Lesley Bechtold) Date: Tue Nov 13 11:28:19 2007 Subject: [Histonet] automated paraffin embedding systems In-Reply-To: <661949901A768E4F9CC16D8AF8F2838CA1C7A8@IS-E2K3.grhs.net> Message-ID: <20071113122632276.00000002152@spikey> Thank you to everyone for so quickly responding!!! I'm calling my Sakura rep for more info. Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 -----Original Message----- From: Mike Pence [mailto:mpence@grhs.net] Sent: Tuesday, November 13, 2007 12:10 PM To: Rene J Buesa; Lesley Bechtold; Histology Network Subject: RE: [Histonet] automated paraffin embedding systems Is anyone out there using this function of the Sakura Xpress? Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, November 13, 2007 11:04 AM To: Lesley Bechtold; Histology Network Subject: Re: [Histonet] automated paraffin embedding systems This embedding instrument is manufactured by Sakura. It is not that the instrument orients the tissue, that is done when the tissues are placed in the cassette and held in place by a plastic frame that will be sectioned along with the tissue when the block is prepared. To function ideally it requires a special type of cassette and cassette holder (that is used alsao by the Sakura Xpress microwave tissue processor). Ren? J. Lesley Bechtold wrote: Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CBraaten <@t> Cheshire-Med.COM Tue Nov 13 11:32:49 2007 From: CBraaten <@t> Cheshire-Med.COM (Christine I. Braaten) Date: Tue Nov 13 11:32:51 2007 Subject: [Histonet] Antibody cocktails Message-ID: Can anybody tell me if there is another company besides Biocare that offers an antibody cocktail with P504S, HMW CK, and p63? Does anybody use the Biocare antibody detection system? I'd appreciate some feedback. Thanks. CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients 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 electronic mail and destroy all copies of the original message. From fudo <@t> ufl.edu Tue Nov 13 11:40:56 2007 From: fudo <@t> ufl.edu (FU,DONGTAO) Date: Tue Nov 13 11:41:03 2007 Subject: [Histonet] staining of negative controls with IHC Message-ID: <1798590190.34131194975656578.JavaMail.osg@osgjas03.cns.ufl.edu> Hi, Ki67 staining in both human and mouse tissue in our lab works very well. We use steamer method 30min in Citra buffer. Ann Dongtao Fu MD, Ph.D Lab Manager Molecular Pathology Core Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 On Tue Nov 13 11:24:00 EST 2007, "Tarango, Mark" wrote: > You probably don't even need to get up to boiling for ki-67. Try > 30 > minutes at 95 C in your pressure cooker. Even on a "negative" > control > you're bound to see something stain positive for ki-67, since > it's a > proliferation marker... although it shouldn't be cytoplasmic. > I'm just > guessing there's got to be a cell or two in the tissue that's > dividing. > > > Mark Adam Tarango HT(ASCP) > Histology & IHC Supervisor > Nevada Cancer Institute > One Breakthrough Way > Las Vegas, NV 89135 > Direct Line (702) 822-5112 > Treo (702) 759-9229 > Fax (702) 939-7663 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > MaryAnn > Dixon > Sent: Tuesday, November 13, 2007 5:18 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] staining of negative controls with IHC > > We are currently retrieving animal tissue in a pressure cooker > with a > program of 125 degrees for 30 secs. This protocol was taken from > biocare but I'm sure this is for human tissue. Most of the tissue > is > retrieved with Biocare's Reveal (pH 6.0). We are receiving some > background staining of our negative controls whereas if we did > not > retrieve them, there is none. We were staining for KI-67 which > is a > nuclear stain. I am using a tonsil for the control. The negative > control > showed some sporatic cytoplasmic staining on the Universal > negative > control from biocare as well as deionized water. Since then we > have > taken the temperature down to 115 degrees and increased the time > to 25 > minutes. It seems to have decreased the cytoplasmic staining but > there > is still some lingering. Anyone out there have any ideas??? I'm > sure > there is a better protocol for retrieving animal tissues. It has > to be a > time and temperature thing! > MaryAnn Dixon > Biological Scientist > Anatomic Pathology > University of Florida > School of Veterinary Medicine > 352-392-2235 ext 4517 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > "EMF " made the following annotations. > ------------------------------------------------------------------------------ > CONFIDENTIALITY NOTICE: This e-mail message, including any > attachments, is for the sole use of the intended recipient(s) and > may contain confidential, proprietary, and/or privileged > information protected by law. If you are not the intended > recipient, you may not use, copy, or distribute this e-mail > message or its attachments. If you believe you have received this > e-mail message in error, please contact the sender by reply > e-mail and destroy all copies of the original message > ============================================================================== > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From godsgalnow <@t> aol.com Tue Nov 13 12:06:20 2007 From: godsgalnow <@t> aol.com (godsgalnow@aol.com) Date: Tue Nov 13 12:06:32 2007 Subject: [Histonet] Antibody cocktails In-Reply-To: References: Message-ID: <8C9F437075F6496-EC4-5B3F@WEBMAIL-MB20.sysops.aol.com> Christine, My understanding is that other vendors do have something similar to the cocktail, but they only contain 2 of the antibodies and the third has to be added at another step, not completely sure on that.? But I do use the BioCare PIN-4 cocktail and have been doing so for 3 years now with no complaints; I?have done?a few thousands of these so far.? I use their Mach 2 Double Stain detection system with the cocktail and it works like a beaut. Roxanne Soto HT(ASCP)QIHC -----Original Message----- From: Christine I. Braaten To: histonet@lists.utsouthwestern.edu Sent: Tue, 13 Nov 2007 12:32 pm Subject: [Histonet] Antibody cocktails Can anybody tell me if there is another company besides Biocare that offers an antibody cocktail with P504S, HMW CK, and p63? Does anybody use the Biocare antibody detection system? I'd appreciate some feedback. Thanks. CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients 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 electronic mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________ Email and AIM finally together. You've gotta check out free AOL Mail! - http://mail.aol.com From katia.catunda <@t> cipax.com.br Tue Nov 13 13:20:18 2007 From: katia.catunda <@t> cipax.com.br (=?iso-8859-1?Q?Ms._K=E1tia_Cristina_Catunda?=) Date: Tue Nov 13 12:20:04 2007 Subject: [Histonet] Histology equipments In-Reply-To: <20071113122632276.00000002152@spikey> Message-ID: Hi there, Beside Sakura, Therm and Mopec, does anybody has anyother manufacturers for histology equipments for recommendation (embedding centers, grossing tables, cassette printers, slide stainers, etc) Tks a lot ? Ms. K?tia Catunda Produ??o +55 12 3203-0612 (direto) +55 12 3203-0633 (PABX) www.cipax.com.br katia.catunda@cipax.com.br From TJJ <@t> Stowers-Institute.org Tue Nov 13 12:31:34 2007 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Tue Nov 13 12:31:51 2007 Subject: [Histonet] Re: staining of negative controls with IHC Message-ID: MaryAnn Dixon wrote: "We are currently retrieving animal tissue in a pressure cooker with a program of 125 degrees for 30 secs. This protocol was taken from biocare but I'm sure this is for human tissue. Most of the tissue is retrieved with Biocare's Reveal (pH 6.0). We are receiving some background staining of our negative controls whereas if we did not retrieve them, there is none. We were staining for KI-67 which is a nuclear stain. I am using a tonsil for the control. The negative control showed some sporatic cytoplasmic staining on the Universal negative control from biocare as well as deionized water. Since then we have taken the temperature down to 115 degrees and increased the time to 25 minutes. It seems to have decreased the cytoplasmic staining but there is still some lingering. Anyone out there have any ideas??? I'm sure there is a better protocol for retrieving animal tissues. It has to be a time and temperature thing!" MaryAnn, you don't mention what type of animal you are staining, and what your IHC protocol looks like. I would never use a universal negative control when doing immunostaining on animals. Use only the Igs from the species the primary antibody was made in. If you are using a monoclonal mouse KI-67, you would need to use non-immune mouse IgGs of the same isotype of your primary antibody. If your antibody is a rabbit polyclonal or monoclonal, you'd need to use non-immune rabbit IgG. It's also important that you match the Ig concentration fo the non-immune (negative) control to the Ig concentration of your primary antibody. If you are using pre-diluted antibodies, you may be using different Ig concentrations. Try an additional run to see if your detection system is causing the staining. Run a tonsil, and do the pressure cooker retrieval as usual, however and omit the primary antibody, using only diluent or buffer in the incubation. Then continue your IHC protocol as usual. If there is still some background/non-specific staining, it's due to your detection system (kit?) and not to the antibody or negative control reagent. Additionally, I never use a universal reagent for detection. Making sure your antibody reactions are appropriate are difficult enough without throwing in additional animal species you don't need. If you still have specific questions, don't hesitate to contact me off-list and I'll be happy to take a look at your protocol and make some recommendations. Good luck! Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 From Dorothy.L.Webb <@t> HealthPartners.Com Tue Nov 13 12:35:36 2007 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Tue Nov 13 12:35:41 2007 Subject: [Histonet] processing Message-ID: <0E394B648E5284478A6CCB78E5AFDA2705635279@hpes1.HealthPartners.int> Now that we can no longer use alcoholic fixatives on breast tissue, is there anyone out there that is putting a slight temperature on the reagents on the processor to enhance fixation and dehydration? I have thought of using stronger alcohol grades and an extra xylene, which would help in the dehydration of the lipids. This would be a dedeicated processor for fatty tissue types only. What is everyone's thoughts and thanks for all input! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From mtarango <@t> nvcancer.org Tue Nov 13 12:42:51 2007 From: mtarango <@t> nvcancer.org (Tarango, Mark) Date: Tue Nov 13 12:50:31 2007 Subject: [Histonet] processing In-Reply-To: <0E394B648E5284478A6CCB78E5AFDA2705635279@hpes1.HealthPartners.int> Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F9C@NVCIEXCH02.NVCI.org> Can you use 20% formalin on breast tissue? Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Webb, Dorothy L Sent: Tuesday, November 13, 2007 10:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing Now that we can no longer use alcoholic fixatives on breast tissue, is there anyone out there that is putting a slight temperature on the reagents on the processor to enhance fixation and dehydration? I have thought of using stronger alcohol grades and an extra xylene, which would help in the dehydration of the lipids. This would be a dedeicated processor for fatty tissue types only. What is everyone's thoughts and thanks for all input! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== From TMcNemar <@t> lmhealth.org Tue Nov 13 13:03:15 2007 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Tue Nov 13 13:03:18 2007 Subject: [Histonet] processing In-Reply-To: <0E394B648E5284478A6CCB78E5AFDA2705635279@hpes1.HealthPartners.int> Message-ID: <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4F4@lmhsmail.lmhealth.org> Why can't you use alcoholic fixatives? Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Webb, Dorothy L Sent: Tuesday, November 13, 2007 1:36 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing Now that we can no longer use alcoholic fixatives on breast tissue, is there anyone out there that is putting a slight temperature on the reagents on the processor to enhance fixation and dehydration? I have thought of using stronger alcohol grades and an extra xylene, which would help in the dehydration of the lipids. This would be a dedeicated processor for fatty tissue types only. What is everyone's thoughts and thanks for all input! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tjasper <@t> copc.net Tue Nov 13 10:13:28 2007 From: tjasper <@t> copc.net (Thomas Jasper) Date: Tue Nov 13 14:13:43 2007 Subject: [Histonet] Histonet down? Message-ID: <90354A475B420441B2A0396E5008D4965E1FF9@copc-sbs.COPC.local> Anyone having trouble with the Histonet? Haven't seen any postings for a while. T. Jasper From rjbuesa <@t> yahoo.com Tue Nov 13 14:40:13 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 13 14:40:19 2007 Subject: [Histonet] Histology equipments In-Reply-To: Message-ID: <9152.47378.qm@web61212.mail.yahoo.com> There are many, but Sakura is the best. Ren? J. "Ms. K?tia Cristina Catunda" wrote: Hi there, Beside Sakura, Therm and Mopec, does anybody has anyother manufacturers for histology equipments for recommendation (embedding centers, grossing tables, cassette printers, slide stainers, etc) Tks a lot Ms. K?tia Catunda Produ??o +55 12 3203-0612 (direto) +55 12 3203-0633 (PABX) www.cipax.com.br katia.catunda@cipax.com.br _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. From barry_m <@t> ozemail.com.au Tue Nov 13 14:58:44 2007 From: barry_m <@t> ozemail.com.au (Barry Madigan) Date: Tue Nov 13 15:01:07 2007 Subject: [Histonet] Antibody cocktails In-Reply-To: References: Message-ID: <000001c82638$012fe800$038fb800$@com.au> Hi Christine, We make up our own antibody cocktail of 34?E12 (DAKO dilution 1:150) and P63 (DAKO 1:400) and stain the section with this cocktail with a Polymer peroxidise detection system (DAB).(Heat retrieval in a high pH retrieval solution for 20 min at 100 C. The stained slides are washed and darkened using a copper sulphate solution. Placed in a buffer wash and then stained for the second time with P504S (AMACR/Racemase) at a dilution of 1:150 with a Polymer Alkaline Phosphatase detection system (Fast Red). We do this all on the BondMax Immunostainer. We have been doing this for over 6 months now and did it this way because we already had all the concentrated antibodies. Proper fixation is vital. Hope this helps. Regards Barry B.Madigan Pathology Queensland Royal Brisbane Campus Australia From eca9 <@t> georgetown.edu Tue Nov 13 15:32:03 2007 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Tue Nov 13 15:32:24 2007 Subject: [Histonet] microtomy technical question Message-ID: <163fae165e00.165e00163fae@imap.georgetown.edu> Hello everyone, I have some questions regarding microtomy. We place our blocks face down on ice. The problem is that some tissues take a very long time before they are ready to be cut(some more than 6 hours). For some tissues like kidney samples (mouse tissue) we have been using glycerol on the ice. This does seem to cut down on the time needed. My question is which other tissues can I use this technique on? Do you have any other suggestions for how to get the tissue hydrated for cutting? Thank you for your help, Eva Permaul Georgetown University From rjbuesa <@t> yahoo.com Tue Nov 13 15:39:58 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 13 15:40:02 2007 Subject: [Histonet] microtomy technical question In-Reply-To: <163fae165e00.165e00163fae@imap.georgetown.edu> Message-ID: <975592.48450.qm@web61220.mail.yahoo.com> Eva: This type of problem (requiring ice, glycerol, and others) to allow sectioning usually stem from either a fixing or processing defficiency. I suggest you to check your fixing times and processing protocols or you will keep having this type of difficulties. Tissues (any type) properly infiltrated (after adequate fixation/processing) do not need anything but a light cooling to allow ribbons to be taken. Ren? J. Eva C Andersson wrote: Hello everyone, I have some questions regarding microtomy. We place our blocks face down on ice. The problem is that some tissues take a very long time before they are ready to be cut(some more than 6 hours). For some tissues like kidney samples (mouse tissue) we have been using glycerol on the ice. This does seem to cut down on the time needed. My question is which other tissues can I use this technique on? Do you have any other suggestions for how to get the tissue hydrated for cutting? Thank you for your help, Eva Permaul Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From gayle.callis <@t> bresnan.net Tue Nov 13 15:57:37 2007 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Tue Nov 13 15:56:50 2007 Subject: [Histonet] It may be more than just a microtomy technical question References: <163fae165e00.165e00163fae@imap.georgetown.edu> Message-ID: <000701c82640$33f907a0$6501a8c0@DHXTS541> Eva, It sounds as though you are processing your tissues too long, and too much water is being removed (over dehydration.) In general, a properly processed mouse tissue after a short schedule will need a bit of a water soak, but not for 6 hours. Another trick is try room temperature or even warm water for a few minutes, then go to ice water, and be careful to NOT cut away what you have soaked - be careful to reapproach the blade in order to cut the sections. Some people have suggested floating blocks face down on a waterbath for a very short time. if you post your processing schedule, Histonetters can help solve some of this problem. Gayle Callis HT,HTL,MT(ASCP) ----- Original Message ----- From: "Eva C Andersson" To: Sent: Tuesday, November 13, 2007 2:32 PM Subject: [Histonet] microtomy technical question > Hello everyone, > I have some questions regarding microtomy. > We place our blocks face down on ice. The problem is that some tissues > take a very long time before they are ready to be cut(some more than 6 > hours). For some tissues like kidney samples (mouse tissue) we have been > using glycerol on the ice. This does seem to cut down on the time needed. > My question is which other tissues can I use this technique on? Do you > have any other suggestions for how to get the tissue hydrated for cutting? > Thank you for your help, > Eva Permaul > Georgetown University > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From schaundrawalton <@t> yahoo.com Tue Nov 13 17:00:06 2007 From: schaundrawalton <@t> yahoo.com (Schaundra Walton) Date: Tue Nov 13 17:00:12 2007 Subject: [Histonet] Giemsa Stain Message-ID: <625759.21602.qm@web58904.mail.re1.yahoo.com> We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. From jqb7 <@t> CDC.GOV Wed Nov 14 04:48:41 2007 From: jqb7 <@t> CDC.GOV (Bartlett, Jeanine (CDC/CCID/NCZVED)) Date: Wed Nov 14 04:49:02 2007 Subject: [Histonet] automated paraffin embedding systems In-Reply-To: <284972.24114.qm@web61211.mail.yahoo.com> References: <20071113110102345.00000002152@spikey> <284972.24114.qm@web61211.mail.yahoo.com> Message-ID: <34BB307EFC9A65429BBB49E330675F72045E21F1@LTA3VS003.ees.hhs.gov> Slight correction. The Xpress microwave tissue processor does not require a special type of cassette. The AutoTEC does, and it can be used on the Xpress or in a traditional processor. But the Xpress can use the same type of plastic cassette used in any traditional processor. Jeanine Bartlett, BS, HT(ASCP)QIHC Centers for Disease Control and Prevention Infectious Diseases Pathology Branch 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, November 13, 2007 12:04 PM To: Lesley Bechtold; Histology Network Subject: Re: [Histonet] automated paraffin embedding systems This embedding instrument is manufactured by Sakura. It is not that the instrument orients the tissue, that is done when the tissues are placed in the cassette and held in place by a plastic frame that will be sectioned along with the tissue when the block is prepared. To function ideally it requires a special type of cassette and cassette holder (that is used alsao by the Sakura Xpress microwave tissue processor). Ren? J. Lesley Bechtold wrote: Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> CDC.GOV Wed Nov 14 04:50:24 2007 From: jqb7 <@t> CDC.GOV (Bartlett, Jeanine (CDC/CCID/NCZVED)) Date: Wed Nov 14 04:50:38 2007 Subject: [Histonet] automated paraffin embedding systems In-Reply-To: <661949901A768E4F9CC16D8AF8F2838CA1C7A8@IS-E2K3.grhs.net> References: <284972.24114.qm@web61211.mail.yahoo.com> <661949901A768E4F9CC16D8AF8F2838CA1C7A8@IS-E2K3.grhs.net> Message-ID: <34BB307EFC9A65429BBB49E330675F72045E21F2@LTA3VS003.ees.hhs.gov> I use both machines. You can use the AutoTEC cassette on any type of processor (including the Xpress) and you do not have to use the AutoTEC and the Xpress both. They can be used independently. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mike Pence Sent: Tuesday, November 13, 2007 12:10 PM To: Rene J Buesa; Lesley Bechtold; Histology Network Subject: RE: [Histonet] automated paraffin embedding systems Is anyone out there using this function of the Sakura Xpress? Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, November 13, 2007 11:04 AM To: Lesley Bechtold; Histology Network Subject: Re: [Histonet] automated paraffin embedding systems This embedding instrument is manufactured by Sakura. It is not that the instrument orients the tissue, that is done when the tissues are placed in the cassette and held in place by a plastic frame that will be sectioned along with the tissue when the block is prepared. To function ideally it requires a special type of cassette and cassette holder (that is used alsao by the Sakura Xpress microwave tissue processor). Ren? J. Lesley Bechtold wrote: Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@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 rjbuesa <@t> yahoo.com Wed Nov 14 07:01:09 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 14 07:01:14 2007 Subject: [Histonet] Giemsa Stain In-Reply-To: <625759.21602.qm@web58904.mail.re1.yahoo.com> Message-ID: <387310.3350.qm@web61217.mail.yahoo.com> Giemsa, as any other stain, is just that, a stain, and stains do not (cannot) have "internal controls". As with any other HC procedure you have to use a positive control along with your tissue. To completely demonstrate that your Giemsa procedure worked as expected, you coul use a blood smear or an appendix section as controls. That will depend on your pathologist's whims. Ren? J. Schaundra Walton wrote: We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. From Terry.Marshall <@t> rothgen.nhs.uk Wed Nov 14 07:22:36 2007 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Wed Nov 14 07:22:43 2007 Subject: [Histonet] Giemsa Stain Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE79@TRFT-EX01.xRothGen.nhs.uk> Sorry Rene, on this occasion I can't agree. Giemsa is not on this occasion being used to bring out granules as it would in it's normal use as a blood/marrow stain. It is just used as a simple quick stain, which will pick up Helicobacter inter alia. If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: 14 November 2007 13:01 To: Schaundra Walton; Histonet Subject: Re: [Histonet] Giemsa Stain Giemsa, as any other stain, is just that, a stain, and stains do not (cannot) have "internal controls". As with any other HC procedure you have to use a positive control along with your tissue. To completely demonstrate that your Giemsa procedure worked as expected, you coul use a blood smear or an appendix section as controls. That will depend on your pathologist's whims. Ren? J. Schaundra Walton wrote: We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed Nov 14 07:41:09 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 14 07:41:15 2007 Subject: [Histonet] Giemsa Stain In-Reply-To: <5C0BED61F529364E86309CADEA63FEF2F3AE79@TRFT-EX01.xRothGen.nhs.uk> Message-ID: <246072.35484.qm@web61218.mail.yahoo.com> I was referring at the fact that IF some coloration took place (of any kind) the procedure worked, and there was a staining. I was not referring to stain specificity. Ren? J. "Marshall Terry Dr, Consultant Histopathologist" wrote: Sorry Rene, on this occasion I can't agree. Giemsa is not on this occasion being used to bring out granules as it would in it's normal use as a blood/marrow stain. It is just used as a simple quick stain, which will pick up Helicobacter inter alia. If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: 14 November 2007 13:01 To: Schaundra Walton; Histonet Subject: Re: [Histonet] Giemsa Stain Giemsa, as any other stain, is just that, a stain, and stains do not (cannot) have "internal controls". As with any other HC procedure you have to use a positive control along with your tissue. To completely demonstrate that your Giemsa procedure worked as expected, you coul use a blood smear or an appendix section as controls. That will depend on your pathologist's whims. Ren? J. Schaundra Walton wrote: We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. From Kemlo.Rogerson <@t> waht.swest.nhs.uk Wed Nov 14 07:42:27 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Wed Nov 14 07:42:34 2007 Subject: [Histonet] Giemsa Stain Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F171@wahtntex2.waht.swest.nhs.uk> "If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose." Terry If it's not blue then it hasn't worked or the organism isn't there! Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From koellingr <@t> comcast.net Wed Nov 14 07:43:25 2007 From: koellingr <@t> comcast.net (koellingr@comcast.net) Date: Wed Nov 14 07:43:35 2007 Subject: [Histonet] Giemsa Stain Message-ID: <111420071343.9538.473AFB7D00086C1A0000254222058863609D09020704040A0105@comcast.net> Rene, Are you saying a histochemical stain for an organism (Helicobactor, AFB, spirochetes, etc) must have a positive control with which I'd agree? But are you saying that for "any" HC stain like elastic, trichrome, PAS, alcian blue, etc that internal controls don't (cannot) even exist? Thanks, Ray Koelling PhenoPath Labs Seattle, WA -------------- Original message -------------- From: Rene J Buesa > Giemsa, as any other stain, is just that, a stain, and stains do not (cannot) > have "internal controls". > As with any other HC procedure you have to use a positive control along with > your tissue. > To completely demonstrate that your Giemsa procedure worked as expected, you > coul use a blood smear or an appendix section as controls. That will depend on > your pathologist's whims. > Ren$B!&(BJ. > > Schaundra Walton wrote: > We've been using a Giemsa stain for our H. pylori cases without a control > tissue. It has come to my attention that this may not be good practice. I was > under the impression that the Giemsa stain had an internal control. Is anyone > else using a this stain for H. pylori? Are you running a tissue control with it? > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Wed Nov 14 07:50:33 2007 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Wed Nov 14 07:50:41 2007 Subject: [Histonet] Giemsa Stain Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE7A@TRFT-EX01.xRothGen.nhs.uk> No. The tissue can still be blue even if the organism isn't there. BTW, has anyone ever seen a Giemsa that doesn't stain? I can't really imagine it. Terry -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@waht.swest.nhs.uk] Sent: 14 November 2007 13:42 To: Marshall Terry Dr, Consultant Histopathologist; Rene J Buesa; Schaundra Walton; Histonet Subject: RE: [Histonet] Giemsa Stain "If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose." Terry If it's not blue then it hasn't worked or the organism isn't there! Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From Terry.Marshall <@t> rothgen.nhs.uk Wed Nov 14 07:53:15 2007 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Wed Nov 14 07:53:21 2007 Subject: [Histonet] Giemsa Stain Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE7B@TRFT-EX01.xRothGen.nhs.uk> So we are singing from the same hymn sheet.... Stains can't have internal controls. Tissues can, in relation to some stains. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of koellingr@comcast.net Sent: 14 November 2007 13:43 To: Rene J Buesa; Schaundra Walton; Histonet Subject: Re: [Histonet] Giemsa Stain Rene, Are you saying a histochemical stain for an organism (Helicobactor, AFB, spirochetes, etc) must have a positive control with which I'd agree? But are you saying that for "any" HC stain like elastic, trichrome, PAS, alcian blue, etc that internal controls don't (cannot) even exist? Thanks, Ray Koelling PhenoPath Labs Seattle, WA -------------- Original message -------------- From: Rene J Buesa > Giemsa, as any other stain, is just that, a stain, and stains do not > (cannot) have "internal controls". > As with any other HC procedure you have to use a positive control > along with your tissue. > To completely demonstrate that your Giemsa procedure worked as > expected, you coul use a blood smear or an appendix section as > controls. That will depend on your pathologist's whims. > Ren$B!&(BJ. > > Schaundra Walton wrote: > We've been using a Giemsa stain for our H. pylori cases without a > control tissue. It has come to my attention that this may not be good > practice. I was under the impression that the Giemsa stain had an > internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@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> waht.swest.nhs.uk Wed Nov 14 07:54:07 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Wed Nov 14 07:54:14 2007 Subject: [Histonet] Giemsa Stain Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F172@wahtntex2.waht.swest.nhs.uk> "No. The tissue can still be blue even if the organism isn't there." Terry I know what you mean; I get days like that too. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From rjbuesa <@t> yahoo.com Wed Nov 14 07:57:03 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 14 07:57:15 2007 Subject: [Histonet] Giemsa Stain In-Reply-To: <5C0BED61F529364E86309CADEA63FEF2F3AE7B@TRFT-EX01.xRothGen.nhs.uk> Message-ID: <588286.70996.qm@web61222.mail.yahoo.com> Amen! Ren? J. "Marshall Terry Dr, Consultant Histopathologist" wrote: So we are singing from the same hymn sheet.... Stains can't have internal controls. Tissues can, in relation to some stains. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of koellingr@comcast.net Sent: 14 November 2007 13:43 To: Rene J Buesa; Schaundra Walton; Histonet Subject: Re: [Histonet] Giemsa Stain Rene, Are you saying a histochemical stain for an organism (Helicobactor, AFB, spirochetes, etc) must have a positive control with which I'd agree? But are you saying that for "any" HC stain like elastic, trichrome, PAS, alcian blue, etc that internal controls don't (cannot) even exist? Thanks, Ray Koelling PhenoPath Labs Seattle, WA -------------- Original message -------------- From: Rene J Buesa > Giemsa, as any other stain, is just that, a stain, and stains do not > (cannot) have "internal controls". > As with any other HC procedure you have to use a positive control > along with your tissue. > To completely demonstrate that your Giemsa procedure worked as > expected, you coul use a blood smear or an appendix section as > controls. That will depend on your pathologist's whims. > Ren$B!&(BJ. > > Schaundra Walton wrote: > We've been using a Giemsa stain for our H. pylori cases without a > control tissue. It has come to my attention that this may not be good > practice. I was under the impression that the Giemsa stain had an > internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. From mpence <@t> grhs.net Wed Nov 14 08:03:00 2007 From: mpence <@t> grhs.net (Mike Pence) Date: Wed Nov 14 08:03:06 2007 Subject: [Histonet] Giemsa Stain In-Reply-To: <625759.21602.qm@web58904.mail.re1.yahoo.com> Message-ID: <661949901A768E4F9CC16D8AF8F2838CA1C7AB@IS-E2K3.grhs.net> We do a Diff Qik stain that takes 30 seconds to do and we use a control with the stain. Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Schaundra Walton Sent: Tuesday, November 13, 2007 5:00 PM To: Histonet Subject: [Histonet] Giemsa Stain We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From plott <@t> uab.edu Wed Nov 14 08:20:59 2007 From: plott <@t> uab.edu (Patricia F Lott) Date: Wed Nov 14 08:21:06 2007 Subject: [Histonet] questions @ MMA Message-ID: <4850136AECAE5447B2E7FF80CA9225300C3557@UABEXMB4.ad.uab.edu> Do you use an adhesive, such as Haupt's, and/or "plus" slides for plastic sections? Also, do you dry them first (after de-plasticization), before starting the stain? Or, should they be re-hydrated? From RSRICHMOND <@t> aol.com Wed Nov 14 08:53:57 2007 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Wed Nov 14 08:54:06 2007 Subject: [Histonet] Re: Giemsa stain Message-ID: The Giemsa stain is widely used as a stain for Helicobacter pylori in gastric biopsy material. It's actually simpler to use a solution of toluidine blue and related dyes in a neutral buffer (such as Diff-Quik II and its many generic equivalents). There is adequate literature to support this technique. It's never good practice to stain for an organism without an appropriate control slide, though in this particular case I don't really think it's necessary from a technical viewpoint - the adjacent tissue provides an adequate control. In the USA, some regulatory agencies (this is a somewhat controversial point) require mention of a control in the pathologist's report. My usual microscopic note reads "a bacterial stain (generic equivalent of Diff-Quik II, with a positive control slide) shows nothing to suggest Helicobacter." If the control slide doesn't contain any bacteria (frequently the case, with histotechs who never look at a slide), I say "suitable" rather than "positive". The control should be a gastric biopsy specimen positive for Helicobacter. The pathologist should inform the histotechs when a suitable case to use as a control comes by. Of course, if you get a gastrectomy specimen done for ulcer disease (shouldn't happen, but it still does occasionally) then you're set for control sections for life. Slides often need to be examined with an oil immersion lens. I do this with all positive specimens, and with apparent negatives when chronic active gastritis (neutrophils infiltrating between the gastric pits) is present. I don't think immunohistochemistry is more sensitive, but reading the slides is an awful lot faster - important if you've got 10 gastric biopsies on your desk. I have no experience with the silver techniques occasionally used. Bob Richmond Samurai Pathologist Knoxville TN From godsgalnow <@t> aol.com Wed Nov 14 09:13:38 2007 From: godsgalnow <@t> aol.com (godsgalnow@aol.com) Date: Wed Nov 14 09:13:51 2007 Subject: [Histonet] research use only Message-ID: <8C9F4E811B87D5F-17E8-97DC@FWM-D32.sysops.aol.com> Hello all.... Can anyone tell me what your protocol is for using RUO's?? Can you bill for them?? How many do you validate? Thanks in advance.... Roxanne ________________________________________________________________________ Email and AIM finally together. You've gotta check out free AOL Mail! - http://mail.aol.com From pierre.chaumat <@t> alphelys.com Wed Nov 14 08:09:44 2007 From: pierre.chaumat <@t> alphelys.com (Pierre CHAUMAT) Date: Wed Nov 14 09:18:58 2007 Subject: [Histonet] RE: Alcoholic fixative on breast tissue ? In-Reply-To: <200711141357.lAEDvSH5003001@smtp11.msg.oleane.net> Message-ID: <0ML21M-1IsIw51qXj-0002EL@mrelayeu.kundenserver.de> Dear Dorothy, I am from France and this is the first time I can read that we can no longer user alcoholc fixatives on breast tissue. Is it a regulatory rule ? Thanks for your help Kind regards --------------Your email -------------------------- Date: Tue, 13 Nov 2007 12:35:36 -0600 From: "Webb, Dorothy L" Subject: [Histonet] processing To: histonet@lists.utsouthwestern.edu Message-ID: <0E394B648E5284478A6CCB78E5AFDA2705635279@hpes1.HealthPartners.int> Content-Type: text/plain; charset="us-ascii" Now that we can no longer use alcoholic fixatives on breast tissue, -----Message d'origine----- De : histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] De la part de histonet-request@lists.utsouthwestern.edu Envoy? : mercredi 14 novembre 2007 14:57 ? : histonet@lists.utsouthwestern.edu Objet : Histonet Digest, Vol 48, Issue 18 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: Antibody cocktails (godsgalnow@aol.com) 2. Histology equipments (Ms. K?tia Cristina Catunda) 3. Re: staining of negative controls with IHC (Johnson, Teri) 4. processing (Webb, Dorothy L) 5. RE: processing (Tarango, Mark) 6. RE: processing (Tom McNemar) 7. Histonet down? (Thomas Jasper) 8. Re: Histology equipments (Rene J Buesa) 9. RE: Antibody cocktails (Barry Madigan) 10. microtomy technical question (Eva C Andersson) 11. Re: microtomy technical question (Rene J Buesa) 12. It may be more than just a microtomy technical question (Gayle Callis) 13. Giemsa Stain (Schaundra Walton) 14. RE: automated paraffin embedding systems (Bartlett, Jeanine (CDC/CCID/NCZVED)) 15. RE: automated paraffin embedding systems (Bartlett, Jeanine (CDC/CCID/NCZVED)) 16. Re: Giemsa Stain (Rene J Buesa) 17. RE: Giemsa Stain (Marshall Terry Dr, Consultant Histopathologist) 18. RE: Giemsa Stain (Rene J Buesa) 19. RE: Giemsa Stain (Kemlo Rogerson) 20. Re: Giemsa Stain (koellingr@comcast.net) 21. RE: Giemsa Stain (Marshall Terry Dr, Consultant Histopathologist) 22. RE: Giemsa Stain (Marshall Terry Dr, Consultant Histopathologist) 23. RE: Giemsa Stain (Kemlo Rogerson) 24. RE: Giemsa Stain (Rene J Buesa) ---------------------------------------------------------------------- Message: 1 Date: Tue, 13 Nov 2007 13:06:20 -0500 From: godsgalnow@aol.com Subject: Re: [Histonet] Antibody cocktails To: CBraaten@Cheshire-Med.COM, histonet@lists.utsouthwestern.edu Message-ID: <8C9F437075F6496-EC4-5B3F@WEBMAIL-MB20.sysops.aol.com> Content-Type: text/plain; charset="us-ascii" Christine, My understanding is that other vendors do have something similar to the cocktail, but they only contain 2 of the antibodies and the third has to be added at another step, not completely sure on that.? But I do use the BioCare PIN-4 cocktail and have been doing so for 3 years now with no complaints; I?have done?a few thousands of these so far.? I use their Mach 2 Double Stain detection system with the cocktail and it works like a beaut. Roxanne Soto HT(ASCP)QIHC -----Original Message----- From: Christine I. Braaten To: histonet@lists.utsouthwestern.edu Sent: Tue, 13 Nov 2007 12:32 pm Subject: [Histonet] Antibody cocktails Can anybody tell me if there is another company besides Biocare that offers an antibody cocktail with P504S, HMW CK, and p63? Does anybody use the Biocare antibody detection system? I'd appreciate some feedback. Thanks. CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients 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 electronic mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________ Email and AIM finally together. You've gotta check out free AOL Mail! - http://mail.aol.com ------------------------------ Message: 2 Date: Tue, 13 Nov 2007 16:20:18 -0300 From: Ms. K?tia Cristina Catunda Subject: [Histonet] Histology equipments To: "'Histology Network'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi there, Beside Sakura, Therm and Mopec, does anybody has anyother manufacturers for histology equipments for recommendation (embedding centers, grossing tables, cassette printers, slide stainers, etc) Tks a lot Ms. Katia Catunda Produgco +55 12 3203-0612 (direto) +55 12 3203-0633 (PABX) www.cipax.com.br katia.catunda@cipax.com.br ------------------------------ Message: 3 Date: Tue, 13 Nov 2007 12:31:34 -0600 From: "Johnson, Teri" Subject: [Histonet] Re: staining of negative controls with IHC To: Message-ID: Content-Type: text/plain; charset="US-ASCII" MaryAnn Dixon wrote: "We are currently retrieving animal tissue in a pressure cooker with a program of 125 degrees for 30 secs. This protocol was taken from biocare but I'm sure this is for human tissue. Most of the tissue is retrieved with Biocare's Reveal (pH 6.0). We are receiving some background staining of our negative controls whereas if we did not retrieve them, there is none. We were staining for KI-67 which is a nuclear stain. I am using a tonsil for the control. The negative control showed some sporatic cytoplasmic staining on the Universal negative control from biocare as well as deionized water. Since then we have taken the temperature down to 115 degrees and increased the time to 25 minutes. It seems to have decreased the cytoplasmic staining but there is still some lingering. Anyone out there have any ideas??? I'm sure there is a better protocol for retrieving animal tissues. It has to be a time and temperature thing!" MaryAnn, you don't mention what type of animal you are staining, and what your IHC protocol looks like. I would never use a universal negative control when doing immunostaining on animals. Use only the Igs from the species the primary antibody was made in. If you are using a monoclonal mouse KI-67, you would need to use non-immune mouse IgGs of the same isotype of your primary antibody. If your antibody is a rabbit polyclonal or monoclonal, you'd need to use non-immune rabbit IgG. It's also important that you match the Ig concentration fo the non-immune (negative) control to the Ig concentration of your primary antibody. If you are using pre-diluted antibodies, you may be using different Ig concentrations. Try an additional run to see if your detection system is causing the staining. Run a tonsil, and do the pressure cooker retrieval as usual, however and omit the primary antibody, using only diluent or buffer in the incubation. Then continue your IHC protocol as usual. If there is still some background/non-specific staining, it's due to your detection system (kit?) and not to the antibody or negative control reagent. Additionally, I never use a universal reagent for detection. Making sure your antibody reactions are appropriate are difficult enough without throwing in additional animal species you don't need. If you still have specific questions, don't hesitate to contact me off-list and I'll be happy to take a look at your protocol and make some recommendations. Good luck! Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 ------------------------------ Message: 4 Date: Tue, 13 Nov 2007 12:35:36 -0600 From: "Webb, Dorothy L" Subject: [Histonet] processing To: histonet@lists.utsouthwestern.edu Message-ID: <0E394B648E5284478A6CCB78E5AFDA2705635279@hpes1.HealthPartners.int> Content-Type: text/plain; charset="us-ascii" Now that we can no longer use alcoholic fixatives on breast tissue, is there anyone out there that is putting a slight temperature on the reagents on the processor to enhance fixation and dehydration? I have thought of using stronger alcohol grades and an extra xylene, which would help in the dehydration of the lipids. This would be a dedeicated processor for fatty tissue types only. What is everyone's thoughts and thanks for all input! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. ------------------------------ Message: 5 Date: Tue, 13 Nov 2007 10:42:51 -0800 From: "Tarango, Mark" Subject: RE: [Histonet] processing To: "Webb, Dorothy L" , histonet@lists.utsouthwestern.edu Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F9C@NVCIEXCH02.NVCI.org> Content-Type: text/plain; charset=us-ascii Can you use 20% formalin on breast tissue? Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Webb, Dorothy L Sent: Tuesday, November 13, 2007 10:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing Now that we can no longer use alcoholic fixatives on breast tissue, is there anyone out there that is putting a slight temperature on the reagents on the processor to enhance fixation and dehydration? I have thought of using stronger alcohol grades and an extra xylene, which would help in the dehydration of the lipids. This would be a dedeicated processor for fatty tissue types only. What is everyone's thoughts and thanks for all input! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ---------------------------------------------------------------------------- -- CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================ == ------------------------------ Message: 6 Date: Tue, 13 Nov 2007 14:03:15 -0500 From: "Tom McNemar" Subject: RE: [Histonet] processing To: "Webb, Dorothy L" , Message-ID: <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4F4@lmhsmail.lmhealth.org> Content-Type: text/plain; charset="iso-8859-1" Why can't you use alcoholic fixatives? Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Webb, Dorothy L Sent: Tuesday, November 13, 2007 1:36 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing Now that we can no longer use alcoholic fixatives on breast tissue, is there anyone out there that is putting a slight temperature on the reagents on the processor to enhance fixation and dehydration? I have thought of using stronger alcohol grades and an extra xylene, which would help in the dehydration of the lipids. This would be a dedeicated processor for fatty tissue types only. What is everyone's thoughts and thanks for all input! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Tue, 13 Nov 2007 08:13:28 -0800 From: "Thomas Jasper" Subject: [Histonet] Histonet down? To: Message-ID: <90354A475B420441B2A0396E5008D4965E1FF9@copc-sbs.COPC.local> Content-Type: text/plain; charset="us-ascii" Anyone having trouble with the Histonet? Haven't seen any postings for a while. T. Jasper ------------------------------ Message: 8 Date: Tue, 13 Nov 2007 12:40:13 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Histology equipments To: "Ms. Katia Cristina Catunda" , 'Histology Network' Message-ID: <9152.47378.qm@web61212.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 There are many, but Sakura is the best. Reni J. "Ms. Katia Cristina Catunda" wrote: Hi there, Beside Sakura, Therm and Mopec, does anybody has anyother manufacturers for histology equipments for recommendation (embedding centers, grossing tables, cassette printers, slide stainers, etc) Tks a lot Ms. Katia Catunda Produgco +55 12 3203-0612 (direto) +55 12 3203-0633 (PABX) www.cipax.com.br katia.catunda@cipax.com.br _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. ------------------------------ Message: 9 Date: Wed, 14 Nov 2007 06:58:44 +1000 From: "Barry Madigan" Subject: RE: [Histonet] Antibody cocktails To: "'Christine I. Braaten'" , Message-ID: <000001c82638$012fe800$038fb800$@com.au> Content-Type: text/plain; charset="iso-8859-1" Hi Christine, We make up our own antibody cocktail of 34_E12 (DAKO dilution 1:150) and P63 (DAKO 1:400) and stain the section with this cocktail with a Polymer peroxidise detection system (DAB).(Heat retrieval in a high pH retrieval solution for 20 min at 100 C. The stained slides are washed and darkened using a copper sulphate solution. Placed in a buffer wash and then stained for the second time with P504S (AMACR/Racemase) at a dilution of 1:150 with a Polymer Alkaline Phosphatase detection system (Fast Red). We do this all on the BondMax Immunostainer. We have been doing this for over 6 months now and did it this way because we already had all the concentrated antibodies. Proper fixation is vital. Hope this helps. Regards Barry B.Madigan Pathology Queensland Royal Brisbane Campus Australia ------------------------------ Message: 10 Date: Tue, 13 Nov 2007 16:32:03 -0500 From: Eva C Andersson Subject: [Histonet] microtomy technical question To: histonet@lists.utsouthwestern.edu Message-ID: <163fae165e00.165e00163fae@imap.georgetown.edu> Content-Type: text/plain; charset=us-ascii Hello everyone, I have some questions regarding microtomy. We place our blocks face down on ice. The problem is that some tissues take a very long time before they are ready to be cut(some more than 6 hours). For some tissues like kidney samples (mouse tissue) we have been using glycerol on the ice. This does seem to cut down on the time needed. My question is which other tissues can I use this technique on? Do you have any other suggestions for how to get the tissue hydrated for cutting? Thank you for your help, Eva Permaul Georgetown University ------------------------------ Message: 11 Date: Tue, 13 Nov 2007 13:39:58 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] microtomy technical question To: Eva C Andersson , histonet@lists.utsouthwestern.edu Message-ID: <975592.48450.qm@web61220.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Eva: This type of problem (requiring ice, glycerol, and others) to allow sectioning usually stem from either a fixing or processing defficiency. I suggest you to check your fixing times and processing protocols or you will keep having this type of difficulties. Tissues (any type) properly infiltrated (after adequate fixation/processing) do not need anything but a light cooling to allow ribbons to be taken. Reni J. Eva C Andersson wrote: Hello everyone, I have some questions regarding microtomy. We place our blocks face down on ice. The problem is that some tissues take a very long time before they are ready to be cut(some more than 6 hours). For some tissues like kidney samples (mouse tissue) we have been using glycerol on the ice. This does seem to cut down on the time needed. My question is which other tissues can I use this technique on? Do you have any other suggestions for how to get the tissue hydrated for cutting? Thank you for your help, Eva Permaul Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. ------------------------------ Message: 12 Date: Tue, 13 Nov 2007 14:57:37 -0700 From: "Gayle Callis" Subject: [Histonet] It may be more than just a microtomy technical question To: "Eva C Andersson" , "Histonet" Message-ID: <000701c82640$33f907a0$6501a8c0@DHXTS541> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Eva, It sounds as though you are processing your tissues too long, and too much water is being removed (over dehydration.) In general, a properly processed mouse tissue after a short schedule will need a bit of a water soak, but not for 6 hours. Another trick is try room temperature or even warm water for a few minutes, then go to ice water, and be careful to NOT cut away what you have soaked - be careful to reapproach the blade in order to cut the sections. Some people have suggested floating blocks face down on a waterbath for a very short time. if you post your processing schedule, Histonetters can help solve some of this problem. Gayle Callis HT,HTL,MT(ASCP) ----- Original Message ----- From: "Eva C Andersson" To: Sent: Tuesday, November 13, 2007 2:32 PM Subject: [Histonet] microtomy technical question > Hello everyone, > I have some questions regarding microtomy. > We place our blocks face down on ice. The problem is that some tissues > take a very long time before they are ready to be cut(some more than 6 > hours). For some tissues like kidney samples (mouse tissue) we have been > using glycerol on the ice. This does seem to cut down on the time needed. > My question is which other tissues can I use this technique on? Do you > have any other suggestions for how to get the tissue hydrated for cutting? > Thank you for your help, > Eva Permaul > Georgetown University > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Tue, 13 Nov 2007 15:00:06 -0800 (PST) From: Schaundra Walton Subject: [Histonet] Giemsa Stain To: Histonet Message-ID: <625759.21602.qm@web58904.mail.re1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. ------------------------------ Message: 14 Date: Wed, 14 Nov 2007 05:48:41 -0500 From: "Bartlett, Jeanine (CDC/CCID/NCZVED)" Subject: RE: [Histonet] automated paraffin embedding systems To: "Rene J Buesa" , "Lesley Bechtold" , "Histology Network" Message-ID: <34BB307EFC9A65429BBB49E330675F72045E21F1@LTA3VS003.ees.hhs.gov> Content-Type: text/plain; charset=iso-8859-1 Slight correction. The Xpress microwave tissue processor does not require a special type of cassette. The AutoTEC does, and it can be used on the Xpress or in a traditional processor. But the Xpress can use the same type of plastic cassette used in any traditional processor. Jeanine Bartlett, BS, HT(ASCP)QIHC Centers for Disease Control and Prevention Infectious Diseases Pathology Branch 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, November 13, 2007 12:04 PM To: Lesley Bechtold; Histology Network Subject: Re: [Histonet] automated paraffin embedding systems This embedding instrument is manufactured by Sakura. It is not that the instrument orients the tissue, that is done when the tissues are placed in the cassette and held in place by a plastic frame that will be sectioned along with the tissue when the block is prepared. To function ideally it requires a special type of cassette and cassette holder (that is used alsao by the Sakura Xpress microwave tissue processor). Reni J. Lesley Bechtold wrote: Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Wed, 14 Nov 2007 05:50:24 -0500 From: "Bartlett, Jeanine (CDC/CCID/NCZVED)" Subject: RE: [Histonet] automated paraffin embedding systems To: "Mike Pence" , "Rene J Buesa" , "Lesley Bechtold" , "Histology Network" Message-ID: <34BB307EFC9A65429BBB49E330675F72045E21F2@LTA3VS003.ees.hhs.gov> Content-Type: text/plain; charset=iso-8859-1 I use both machines. You can use the AutoTEC cassette on any type of processor (including the Xpress) and you do not have to use the AutoTEC and the Xpress both. They can be used independently. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mike Pence Sent: Tuesday, November 13, 2007 12:10 PM To: Rene J Buesa; Lesley Bechtold; Histology Network Subject: RE: [Histonet] automated paraffin embedding systems Is anyone out there using this function of the Sakura Xpress? Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, November 13, 2007 11:04 AM To: Lesley Bechtold; Histology Network Subject: Re: [Histonet] automated paraffin embedding systems This embedding instrument is manufactured by Sakura. It is not that the instrument orients the tissue, that is done when the tissues are placed in the cassette and held in place by a plastic frame that will be sectioned along with the tissue when the block is prepared. To function ideally it requires a special type of cassette and cassette holder (that is used alsao by the Sakura Xpress microwave tissue processor). Reni J. Lesley Bechtold wrote: Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Wed, 14 Nov 2007 05:01:09 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Giemsa Stain To: Schaundra Walton , Histonet Message-ID: <387310.3350.qm@web61217.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Giemsa, as any other stain, is just that, a stain, and stains do not (cannot) have "internal controls". As with any other HC procedure you have to use a positive control along with your tissue. To completely demonstrate that your Giemsa procedure worked as expected, you coul use a blood smear or an appendix section as controls. That will depend on your pathologist's whims. Reni J. Schaundra Walton wrote: We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. ------------------------------ Message: 17 Date: Wed, 14 Nov 2007 13:22:36 -0000 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Giemsa Stain To: "Rene J Buesa" , "Schaundra Walton" , "Histonet" Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE79@TRFT-EX01.xRothGen.nhs.uk> Content-Type: text/plain; charset="iso-8859-1" Sorry Rene, on this occasion I can't agree. Giemsa is not on this occasion being used to bring out granules as it would in it's normal use as a blood/marrow stain. It is just used as a simple quick stain, which will pick up Helicobacter inter alia. If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: 14 November 2007 13:01 To: Schaundra Walton; Histonet Subject: Re: [Histonet] Giemsa Stain Giemsa, as any other stain, is just that, a stain, and stains do not (cannot) have "internal controls". As with any other HC procedure you have to use a positive control along with your tissue. To completely demonstrate that your Giemsa procedure worked as expected, you coul use a blood smear or an appendix section as controls. That will depend on your pathologist's whims. Reni J. Schaundra Walton wrote: We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Wed, 14 Nov 2007 05:41:09 -0800 (PST) From: Rene J Buesa Subject: RE: [Histonet] Giemsa Stain To: "Marshall Terry Dr, Consultant Histopathologist" , Schaundra Walton , Histonet Message-ID: <246072.35484.qm@web61218.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I was referring at the fact that IF some coloration took place (of any kind) the procedure worked, and there was a staining. I was not referring to stain specificity. Reni J. "Marshall Terry Dr, Consultant Histopathologist" wrote: Sorry Rene, on this occasion I can't agree. Giemsa is not on this occasion being used to bring out granules as it would in it's normal use as a blood/marrow stain. It is just used as a simple quick stain, which will pick up Helicobacter inter alia. If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: 14 November 2007 13:01 To: Schaundra Walton; Histonet Subject: Re: [Histonet] Giemsa Stain Giemsa, as any other stain, is just that, a stain, and stains do not (cannot) have "internal controls". As with any other HC procedure you have to use a positive control along with your tissue. To completely demonstrate that your Giemsa procedure worked as expected, you coul use a blood smear or an appendix section as controls. That will depend on your pathologist's whims. Reni J. Schaundra Walton wrote: We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. ------------------------------ Message: 19 Date: Wed, 14 Nov 2007 13:42:27 -0000 From: "Kemlo Rogerson" Subject: RE: [Histonet] Giemsa Stain To: "Marshall Terry Dr, Consultant Histopathologist" , "Rene J Buesa" , "Schaundra Walton" , "Histonet" Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F171@wahtntex2.waht.swest.nhs.uk> Content-Type: text/plain; charset="us-ascii" "If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose." Terry If it's not blue then it hasn't worked or the organism isn't there! Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation ------------------------------ Message: 20 Date: Wed, 14 Nov 2007 13:43:25 +0000 From: koellingr@comcast.net Subject: Re: [Histonet] Giemsa Stain To: Rene J Buesa , Schaundra Walton , Histonet Message-ID: <111420071343.9538.473AFB7D00086C1A0000254222058863609D09020704040A0105@comc ast.net> Content-Type: text/plain Rene, Are you saying a histochemical stain for an organism (Helicobactor, AFB, spirochetes, etc) must have a positive control with which I'd agree? But are you saying that for "any" HC stain like elastic, trichrome, PAS, alcian blue, etc that internal controls don't (cannot) even exist? Thanks, Ray Koelling PhenoPath Labs Seattle, WA -------------- Original message -------------- From: Rene J Buesa > Giemsa, as any other stain, is just that, a stain, and stains do not (cannot) > have "internal controls". > As with any other HC procedure you have to use a positive control along with > your tissue. > To completely demonstrate that your Giemsa procedure worked as expected, you > coul use a blood smear or an appendix section as controls. That will depend on > your pathologist's whims. > Ren$B!&(BJ. > > Schaundra Walton wrote: > We've been using a Giemsa stain for our H. pylori cases without a control > tissue. It has come to my attention that this may not be good practice. I was > under the impression that the Giemsa stain had an internal control. Is anyone > else using a this stain for H. pylori? Are you running a tissue control with it? > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: Wed, 14 Nov 2007 13:50:33 -0000 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Giemsa Stain To: "Kemlo Rogerson" , "Rene J Buesa" , "Schaundra Walton" , "Histonet" Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE7A@TRFT-EX01.xRothGen.nhs.uk> Content-Type: text/plain; charset="us-ascii" No. The tissue can still be blue even if the organism isn't there. BTW, has anyone ever seen a Giemsa that doesn't stain? I can't really imagine it. Terry -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@waht.swest.nhs.uk] Sent: 14 November 2007 13:42 To: Marshall Terry Dr, Consultant Histopathologist; Rene J Buesa; Schaundra Walton; Histonet Subject: RE: [Histonet] Giemsa Stain "If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose." Terry If it's not blue then it hasn't worked or the organism isn't there! Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation ------------------------------ Message: 22 Date: Wed, 14 Nov 2007 13:53:15 -0000 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Giemsa Stain To: , "Rene J Buesa" , "Schaundra Walton" , "Histonet" Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE7B@TRFT-EX01.xRothGen.nhs.uk> Content-Type: text/plain; charset="us-ascii" So we are singing from the same hymn sheet.... Stains can't have internal controls. Tissues can, in relation to some stains. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of koellingr@comcast.net Sent: 14 November 2007 13:43 To: Rene J Buesa; Schaundra Walton; Histonet Subject: Re: [Histonet] Giemsa Stain Rene, Are you saying a histochemical stain for an organism (Helicobactor, AFB, spirochetes, etc) must have a positive control with which I'd agree? But are you saying that for "any" HC stain like elastic, trichrome, PAS, alcian blue, etc that internal controls don't (cannot) even exist? Thanks, Ray Koelling PhenoPath Labs Seattle, WA -------------- Original message -------------- From: Rene J Buesa > Giemsa, as any other stain, is just that, a stain, and stains do not > (cannot) have "internal controls". > As with any other HC procedure you have to use a positive control > along with your tissue. > To completely demonstrate that your Giemsa procedure worked as > expected, you coul use a blood smear or an appendix section as > controls. That will depend on your pathologist's whims. > Ren$B!&(BJ. > > Schaundra Walton wrote: > We've been using a Giemsa stain for our H. pylori cases without a > control tissue. It has come to my attention that this may not be good > practice. I was under the impression that the Giemsa stain had an > internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@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: 23 Date: Wed, 14 Nov 2007 13:54:07 -0000 From: "Kemlo Rogerson" Subject: RE: [Histonet] Giemsa Stain To: "Marshall Terry Dr, Consultant Histopathologist" , "Rene J Buesa" , "Schaundra Walton" , "Histonet" Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F172@wahtntex2.waht.swest.nhs.uk> Content-Type: text/plain; charset="us-ascii" "No. The tissue can still be blue even if the organism isn't there." Terry I know what you mean; I get days like that too. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation ------------------------------ Message: 24 Date: Wed, 14 Nov 2007 05:57:03 -0800 (PST) From: Rene J Buesa Subject: RE: [Histonet] Giemsa Stain To: "Marshall Terry Dr, Consultant Histopathologist" , koellingr@comcast.net, Schaundra Walton , Histonet Message-ID: <588286.70996.qm@web61222.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Amen! Reni J. "Marshall Terry Dr, Consultant Histopathologist" wrote: So we are singing from the same hymn sheet.... Stains can't have internal controls. Tissues can, in relation to some stains. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of koellingr@comcast.net Sent: 14 November 2007 13:43 To: Rene J Buesa; Schaundra Walton; Histonet Subject: Re: [Histonet] Giemsa Stain Rene, Are you saying a histochemical stain for an organism (Helicobactor, AFB, spirochetes, etc) must have a positive control with which I'd agree? But are you saying that for "any" HC stain like elastic, trichrome, PAS, alcian blue, etc that internal controls don't (cannot) even exist? Thanks, Ray Koelling PhenoPath Labs Seattle, WA -------------- Original message -------------- From: Rene J Buesa > Giemsa, as any other stain, is just that, a stain, and stains do not > (cannot) have "internal controls". > As with any other HC procedure you have to use a positive control > along with your tissue. > To completely demonstrate that your Giemsa procedure worked as > expected, you coul use a blood smear or an appendix section as > controls. That will depend on your pathologist's whims. > Ren$B!&(BJ. > > Schaundra Walton wrote: > We've been using a Giemsa stain for our H. pylori cases without a > control tissue. It has come to my attention that this may not be good > practice. I was under the impression that the Giemsa stain had an > internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 18 **************************************** From JWEEMS <@t> sjha.org Wed Nov 14 09:19:01 2007 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Nov 14 09:19:28 2007 Subject: [Histonet] research use only In-Reply-To: <8C9F4E811B87D5F-17E8-97DC@FWM-D32.sysops.aol.com> Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD32048F422E@sjhaexc02.sjha.org> According to FDA they can not be billed. We do not bill and include a disclaimer (required by CAP) in the report. j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of godsgalnow@aol.com Sent: Wednesday, November 14, 2007 10:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] research use only Hello all.... Can anyone tell me what your protocol is for using RUO's?? Can you bill for them?? How many do you validate? Thanks in advance.... Roxanne ________________________________________________________________________ Email and AIM finally together. You've gotta check out free AOL Mail! - http://mail.aol.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From Terry.Marshall <@t> rothgen.nhs.uk Wed Nov 14 09:20:33 2007 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Wed Nov 14 09:20:43 2007 Subject: [Histonet] Re: Giemsa stain Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE7C@TRFT-EX01.xRothGen.nhs.uk> Bob, always a great pleasure to read your wisdom. Here though, you seem to be blowing hot then cold about a control for Giemsa for Helicobacter. Here's the crux - will you ever see a situation in which the tissue is stained and the bacteria not? I don't think so. Terry PS All stains work, Immuno is the most sensitive and easy to read, but not of course to do. The "blue" stains tend to be overdone. Silver stains coat the organisms and make them look as if they are wearing a woolly jumper! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert Richmond Sent: 14 November 2007 14:54 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Giemsa stain The Giemsa stain is widely used as a stain for Helicobacter pylori in gastric biopsy material. It's actually simpler to use a solution of toluidine blue and related dyes in a neutral buffer (such as Diff-Quik II and its many generic equivalents). There is adequate literature to support this technique. It's never good practice to stain for an organism without an appropriate control slide, though in this particular case I don't really think it's necessary from a technical viewpoint - the adjacent tissue provides an adequate control. In the USA, some regulatory agencies (this is a somewhat controversial point) require mention of a control in the pathologist's report. My usual microscopic note reads "a bacterial stain (generic equivalent of Diff-Quik II, with a positive control slide) shows nothing to suggest Helicobacter." If the control slide doesn't contain any bacteria (frequently the case, with histotechs who never look at a slide), I say "suitable" rather than "positive". The control should be a gastric biopsy specimen positive for Helicobacter. The pathologist should inform the histotechs when a suitable case to use as a control comes by. Of course, if you get a gastrectomy specimen done for ulcer disease (shouldn't happen, but it still does occasionally) then you're set for control sections for life. Slides often need to be examined with an oil immersion lens. I do this with all positive specimens, and with apparent negatives when chronic active gastritis (neutrophils infiltrating between the gastric pits) is present. I don't think immunohistochemistry is more sensitive, but reading the slides is an awful lot faster - important if you've got 10 gastric biopsies on your desk. I have no experience with the silver techniques occasionally used. Bob Richmond Samurai Pathologist Knoxville TN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From wia2005 <@t> med.cornell.edu Wed Nov 14 09:57:30 2007 From: wia2005 <@t> med.cornell.edu (William Ares) Date: Wed Nov 14 09:57:40 2007 Subject: [Histonet] PCNA Message-ID: <2b2c3b7d1ac2.473ad49a@med.cornell.edu> hi histonetters: the PCNA protocol listed on ihcworld (http://www.ihcworld.com/_protocols/antibody_protocols/pcna_santa_cruz.htm) doesn't mention what secondary antibody to use when doing PCNA staining. does anyone know the recommended secondary? also, does anyone have a better/different protocol for PCNA? I'm using Calbiochem's mouse anti-PCNA. Thanks for the help. William Ares Research Technician Laboratory of Molecular and Developmental Neuroscience Weill Medical College of Cornell University Tel: (212) 746 5056 Email: wia2005@med.cornell.edu From gentras <@t> vetmed.auburn.edu Wed Nov 14 10:13:10 2007 From: gentras <@t> vetmed.auburn.edu (Atoska Gentry) Date: Wed Nov 14 10:13:21 2007 Subject: [Histonet] clearing agent Message-ID: <473B1E96.4050608@vetmed.auburn.edu> hello, has anyone found a tried & proven comparable replacement clearing agent for Fisher's discontinued Tissue Clear III # SH3-4? Your prompt replies will be much appreciated. Atoska -- Atoska S. Gentry, B.S., HT(ASCP) Research Assistant IV Scott-Ritchey RSCH Center College of Vet. Med Auburn, AL 36849 PH (334) 844-5579 FAX (334) 844-5850 email: gentras@vetmed.auburn.edu From shive003 <@t> umn.edu Wed Nov 14 10:41:03 2007 From: shive003 <@t> umn.edu (Jan Shivers) Date: Wed Nov 14 10:41:17 2007 Subject: [Histonet] Animal IHC - BVDv Ab source Message-ID: <00bf01c826dd$2565b6f0$a1065486@auxs.umn.edu> Could anyone please forward me their BVD (Bovine Viral Diarrhea virus) antibody vendor name? My previous vendor no longer supplies it, and I'm not happy with my new source (nonspecific background issues). I'd appreciate any information you may be able to provide. I need to find a new vendor as soon as possible. Thanks much. Jan Shivers UMN Vet Diag Lab St. Paul, MN shive003@umn.edu From Robert.Lott <@t> TriadHospitals.com Wed Nov 14 11:15:59 2007 From: Robert.Lott <@t> TriadHospitals.com (Lott, Robert) Date: Wed Nov 14 11:17:03 2007 Subject: [Histonet] Plastic Sections Message-ID: <518A08C53ED96D419F498D309E64A36A3E9457@CPRTEVS03.triadhospitals.net> Do you use a slide adhesive, such as Haupt's, and/or "plus" slides for plastic sections? Also, do you dry them first (after de-plasticization), before starting the stain? Or, should it be re-hydrated? Patty F. Lott, HT(ASCP) Orthopedic Research Laboratory University of Alabama in Birmingham From Maxim_71 <@t> mail.ru Wed Nov 14 11:46:56 2007 From: Maxim_71 <@t> mail.ru (Maxim Peshkov) Date: Wed Nov 14 11:47:48 2007 Subject: [Histonet] processing Message-ID: <1311885352.20071114204656@mail.ru> Dorothy: We always fixes our breast specimens with 10% formalin and earlier have had great troubles in processing and cutting. When we started use acetone as dehydratant and mixtures of aliphatic hydrocarbons (C7-C10) as clearant, then we got much better results than earlier, but some difficultes in cut still here. Now we use isopropanol as dehydratant and mineral oil as clearant and have best results without any problems. We process all our tissues manually and simultaneously with all specimens, including curretage, bone, uterus, skin etc. Our pathologists and pathologists from other hospitals like these slides. Next year we will be do IHC for breast. Sincerely, Maxim Peshkov Russia, Taganrog. From schaundrawalton <@t> yahoo.com Wed Nov 14 11:53:33 2007 From: schaundrawalton <@t> yahoo.com (Schaundra Walton) Date: Wed Nov 14 11:53:36 2007 Subject: [Histonet] Re: Giemsa Stain Message-ID: <162963.24543.qm@web58906.mail.re1.yahoo.com> Oops my mistake. I understand that stains themselves can't have internal controls. I meant to say that I was under the impression that the tissue had an internal control. In other words I thought that the nuclei stained blue in addition to the H. pylori organism. Therefore, the nuclei served as an internal control. We've been running this stain without a control tissue for quite some time. We've never been cited on a CAP inspection for lack of a control which is why I was curious what other people were doing. One of our pathologists was concerned. Thanks for the advise. Schaundra Walton BS HTL(ASCP) Swedish American Hospital 1401 E. State St. Rockford, IL 61104 --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From Terry.Marshall <@t> rothgen.nhs.uk Wed Nov 14 11:53:34 2007 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Wed Nov 14 11:53:41 2007 Subject: [Histonet] processing Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE80@TRFT-EX01.xRothGen.nhs.uk> Maxim, it's hard to credit that slides are still processed manually. How many in a day? Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Maxim Peshkov Sent: 14 November 2007 17:47 To: Dorothy.L.Webb@HealthPartners.Com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] processing Dorothy: We always fixes our breast specimens with 10% formalin and earlier have had great troubles in processing and cutting. When we started use acetone as dehydratant and mixtures of aliphatic hydrocarbons (C7-C10) as clearant, then we got much better results than earlier, but some difficultes in cut still here. Now we use isopropanol as dehydratant and mineral oil as clearant and have best results without any problems. We process all our tissues manually and simultaneously with all specimens, including curretage, bone, uterus, skin etc. Our pathologists and pathologists from other hospitals like these slides. Next year we will be do IHC for breast. Sincerely, Maxim Peshkov Russia, Taganrog. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mtarango <@t> nvcancer.org Wed Nov 14 11:53:01 2007 From: mtarango <@t> nvcancer.org (Tarango, Mark) Date: Wed Nov 14 11:55:35 2007 Subject: [Histonet] Giemsa Stain In-Reply-To: <5C0BED61F529364E86309CADEA63FEF2F3AE7A@TRFT-EX01.xRothGen.nhs.uk> Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6FA5@NVCIEXCH02.NVCI.org> Shit! I can't get my giemsas to stain! Seriously, if you're looking for H. Pylori, you should have an H. Pylori control. If you are doing a giemsa on a blood smear, then you should have WBCs there to stain all those pretty colors (in which I can't see why you'd need an H. Pylori control). It depends on the reason you're doing the stain. Looking for H. Pylori this way seems old school anyway. I'd rather go to a hospital that checks by IHC or alcian yellow. I like what I'm staining to stand out. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr, Consultant Histopathologist Sent: Wednesday, November 14, 2007 5:51 AM To: Kemlo Rogerson; Rene J Buesa; Schaundra Walton; Histonet Subject: RE: [Histonet] Giemsa Stain No. The tissue can still be blue even if the organism isn't there. BTW, has anyone ever seen a Giemsa that doesn't stain? I can't really imagine it. Terry -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@waht.swest.nhs.uk] Sent: 14 November 2007 13:42 To: Marshall Terry Dr, Consultant Histopathologist; Rene J Buesa; Schaundra Walton; Histonet Subject: RE: [Histonet] Giemsa Stain "If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose." Terry If it's not blue then it hasn't worked or the organism isn't there! Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== From tjasper <@t> copc.net Wed Nov 14 12:13:25 2007 From: tjasper <@t> copc.net (Thomas Jasper) Date: Wed Nov 14 12:13:37 2007 Subject: [Histonet] Registry Exam Materials Message-ID: <90354A475B420441B2A0396E5008D4965E1FFE@copc-sbs.COPC.local> Hello Histofolk, I've got a temp working here preparing for the registry exam. Any recommendations for study materials? Preferably something that's inexpensive and easily obtained. Thanks, Tom J. Thomas Jasper HT (ASCP) BAS Histology Supervisor Central Oregon Regional Pathology Services Bend, Oregon 97701 tjasper@copc.net 541/693-2677 ext. 4050 From Maxim_71 <@t> mail.ru Wed Nov 14 12:24:06 2007 From: Maxim_71 <@t> mail.ru (Maxim Peshkov) Date: Wed Nov 14 12:25:38 2007 Subject: [Histonet] processing In-Reply-To: <5C0BED61F529364E86309CADEA63FEF2F3AE80@TRFT-EX01.xRothGen.nhs.uk> References: <5C0BED61F529364E86309CADEA63FEF2F3AE80@TRFT-EX01.xRothGen.nhs.uk> Message-ID: <1805107028.20071114212406@mail.ru> Terry: We prepare daily approximate from 250 up to 450 slides. Our H&E staining and coverslipping do also manually. It is very bored. Sincerely, Maxim Peshkov Russia, Taganrog. From RSRICHMOND <@t> aol.com Wed Nov 14 12:32:57 2007 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Wed Nov 14 12:33:03 2007 Subject: [Histonet] Re: processing Message-ID: Neutral buffered formalin is the prescribed fixative for breast tissue .for the USA's FDA approved HER2 immunostaining procedure. Neutral buffered formalin is a quite adequate fixative for fatty breast tissue as long as you do two things: 1. The pathologist has to cut the damn tissue thin enough, and not overload the cassettes. I'm appalled at the number of pathologists who are unable or unwilling to do this. I think this detail is particularly important for manual processing (such as our correspondent in Russia describes), though probably no one now working in the USA remembers this technique, since the Technicon processor came into use in the early 1930's - probably the most revolutionary change in surgical pathology in the 20th century, with the possible exception of the cryostat (refrigerated microtome) in the early 1960's. 2. Breast tissue needs to fix overnight. This prolonged fixation is also prescribed by the FDA for the HER2 IHC. You have to sympathize a lot with the patient's anxiety about the outcome of her biopsy, but she needs the right diagnosis, not the fast diagnosis. Pathologists need to remind their surgeons of this delay from time to time (so I learned from Bill Shelley of blest memory, my first teacher in surgical pathology, at Johns Hopkins around 1970). Unfortunately, the commercial labs we compete with are bound by none of these injunctions - they can promise you the moon and then moon you - On another topic today, Mark Adam Tarango in Las Vegas refers to my advocacy of Diff-Quik II as "old school". Does that make me feel old!, since I set the technique up in a number of labs in the early 1990's. When this old boy was in school, Helicobacter (let alone staining it) lay even further in the future than the CT scanner. Bob Richmond Samurai Pathologist Knoxville TN From pruegg <@t> ihctech.net Wed Nov 14 12:40:41 2007 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Wed Nov 14 12:41:05 2007 Subject: [Histonet] glycosaminoglycan (GAG). Message-ID: <006b01c826ed$e0512ca0$6801a8c0@Patsy> Does anyone know of a specific marker for this, antibody or special stain? I know we have used saffrinin o, azure b at different ph's and maybe some collagen antibodies in my past life working in bone, but maybe there is something else out there I have missed? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC 12635 Montview Blvd. Ste.215 Aurora, Colorado 80045 Phone: 720-859-4060 Fax: 720-859-4110 pruegg@ihctech.net www.ihctech.net www.ihcrg.org From kfineout <@t> hotmail.com Wed Nov 14 12:50:39 2007 From: kfineout <@t> hotmail.com (Kelly Larson) Date: Wed Nov 14 12:50:52 2007 Subject: [Histonet] Clearing Agent Message-ID: We use Protocol Define for our stain clarifier and it works great:) _________________________________________________________________ Boo!?Scare away worms, viruses and so much more! Try Windows Live OneCare! http://onecare.live.com/standard/en-us/purchase/trial.aspx?s_cid=wl_hotmailnews From kfineout <@t> hotmail.com Wed Nov 14 12:52:45 2007 From: kfineout <@t> hotmail.com (Kelly Larson) Date: Wed Nov 14 12:52:49 2007 Subject: [Histonet] Breast Processing Message-ID: We fix all specimens, including breast, in 10% NBF and run all specimens on the same processor (same program) at 40?C for all stations up to the paraffin. This works well for us as long as the sections are cut nicely. Hope this helps!! _________________________________________________________________ Climb to the top of the charts!? Play Star Shuffle:? the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct From histoinfo <@t> comcast.net Wed Nov 14 12:56:12 2007 From: histoinfo <@t> comcast.net (histoinfo@comcast.net) Date: Wed Nov 14 12:56:18 2007 Subject: [Histonet] It may be more than just a microtomy technical question Message-ID: <111420071856.27775.473B44CC00031AED00006C7F221559341401000207019B9C0708@comcast.net> This is going to sound strange but try it, it really works. Soak your blocks in Group B strep btoth a liquid media used in Microbiology. You can use the glass lid to a staining dish and soak about 6 blicks at a time. They only need to soak a few minutes. This works great on tissue like liver & kidney but I have used it on everything. I even soaked all my registry blocks using this method years ago. Jennifer Saunders HT (ASCP) From hej01 <@t> health.state.ny.us Wed Nov 14 12:59:43 2007 From: hej01 <@t> health.state.ny.us (Helen E Johnson) Date: Wed Nov 14 12:59:56 2007 Subject: [Histonet] slide & cassette printers Message-ID: Hi Histonetters, Which slide & cassette printer would you recommend? Helen Johnson (hej01@health.state.ny.us) IMPORTANT NOTICE: This e-mail and any attachments may contain confidential or sensitive information which is, or may be, legally privileged or otherwise protected by law from further disclosure. It is intended only for the addressee. If you received this in error or from someone who was not authorized to send it to you, please do not distribute, copy or use it or any attachments. Please notify the sender immediately by reply e-mail and delete this from your system. Thank you for your cooperation. From sjchtascp <@t> yahoo.com Wed Nov 14 13:10:31 2007 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Wed Nov 14 13:10:35 2007 Subject: [Histonet] Seeking an HT position Message-ID: <315806.27443.qm@web38215.mail.mud.yahoo.com> I'm looking for an HT position in the Madison Wisconsin or surrounding area. I'd really appreciate any leands anyone might have. Thanks much all, Steve --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. From mcauliff <@t> umdnj.edu Wed Nov 14 13:31:24 2007 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Wed Nov 14 13:31:46 2007 Subject: [Histonet] glycosaminoglycan (GAG). In-Reply-To: <006b01c826ed$e0512ca0$6801a8c0@Patsy> References: <006b01c826ed$e0512ca0$6801a8c0@Patsy> Message-ID: <473B4D0C.5090403@umdnj.edu> Hi Patsy: Alcian Blue (1% AB in 3% acetic acid) at pH 2.5 will stain both carboxylated and sulfated GAG. At pH 1.0 only sulfated GAG will stain. Geoff Patsy Ruegg wrote: > Does anyone know of a specific marker for this, antibody or special stain? > I know we have used saffrinin o, azure b at different ph's and maybe some > collagen antibodies in my past life working in bone, but maybe there is > something else out there I have missed? > > > > Patsy > > > > > > Patsy Ruegg, HT(ASCP)QIHC > > IHCtech, LLC > > 12635 Montview Blvd. Ste.215 > > Aurora, Colorado 80045 > > Phone: 720-859-4060 > > Fax: 720-859-4110 > > pruegg@ihctech.net > > www.ihctech.net > > www.ihcrg.org > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcauliff@umdnj.edu ********************************************** From AGrobe2555 <@t> aol.com Wed Nov 14 14:07:39 2007 From: AGrobe2555 <@t> aol.com (AGrobe2555@aol.com) Date: Wed Nov 14 14:07:52 2007 Subject: [Histonet] glycosaminoglycan (GAG). Message-ID: Geoff, Would you mind posting a more detailed protocol for this staining. I will also need to do this on samples we have in the lab. Thanks, Albert Albert C. Grobe, PhD International Heart Institute of Montana Foundation Tissue Engineering Lab, Saint Patrick Hospital ************************************** See what's new at http://www.aol.com From sbreeden <@t> nmda.nmsu.edu Wed Nov 14 14:26:20 2007 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Wed Nov 14 14:26:34 2007 Subject: [Histonet] BVD Antibody source Message-ID: <4D14F0FC9316DD41972D5F03C070908B8F4755@nmdamailsvr.nmda.ad.nmsu.edu> Jan, I got my antibody from Oklahoma State University, OK Animal Disease Diagnostic Laboratory. Phone is 405-744-6623 (I believe that's the direct line into the lab). Good clean slides using this antibody! Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 From pruegg <@t> ihctech.net Wed Nov 14 15:17:59 2007 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Wed Nov 14 15:18:14 2007 Subject: [Histonet] glycosaminoglycan (GAG). In-Reply-To: <473B4D0C.5090403@umdnj.edu> Message-ID: <007f01c82703$d59f2120$6801a8c0@Patsy> Thanks Geoff, Do you think formic acid decal will have any adverse effect on the ffpe samples I want to do this on? Patsy -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Wednesday, November 14, 2007 12:31 PM To: Patsy Ruegg Cc: 'Histonet' Subject: Re: [Histonet] glycosaminoglycan (GAG). Hi Patsy: Alcian Blue (1% AB in 3% acetic acid) at pH 2.5 will stain both carboxylated and sulfated GAG. At pH 1.0 only sulfated GAG will stain. Geoff Patsy Ruegg wrote: > Does anyone know of a specific marker for this, antibody or special stain? > I know we have used saffrinin o, azure b at different ph's and maybe some > collagen antibodies in my past life working in bone, but maybe there is > something else out there I have missed? > > > > Patsy > > > > > > Patsy Ruegg, HT(ASCP)QIHC > > IHCtech, LLC > > 12635 Montview Blvd. Ste.215 > > Aurora, Colorado 80045 > > Phone: 720-859-4060 > > Fax: 720-859-4110 > > pruegg@ihctech.net > > www.ihctech.net > > www.ihcrg.org > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcauliff@umdnj.edu ********************************************** From Heather.D.Renko <@t> osfhealthcare.org Wed Nov 14 15:27:18 2007 From: Heather.D.Renko <@t> osfhealthcare.org (Renko, Heather D.) Date: Wed Nov 14 15:27:47 2007 Subject: [Histonet] RE: positive controls-CAP Speak Message-ID: <40026EDDE64CDA47AB382C52619ACD3C0A09DBFC@pmc-rfd-mx01.intranet.osfnet.org> Here is the latest and greatest on positive controls in relation to the special stains in a CAP accredited laboratory; ANP.21400 Phase II N/A YES NO Are positive controls run routinely on all special stains, with reactivity results documented, and are they verified for acceptability before reporting results? NOTE: A positive control slide must be run at the same time as any single or group of slides stained with the same special stain. The tissue chosen for the special stain control slide must be appropriate in type and amount. Both the control slide and the test tissue slide must be judged technically acceptable before the results of the special stains are reported. Sept. 2007 Heather D. Renko, Histology Coordinator OSF Saint Anthony Medical Center 5666 East State Street Rockford, Illinois 61108 Direct: 815-395-5410 Heather.D.Renko@osfhealthcare.org ============================================================================== The information in this message is confidential and may be legally privileged. Access to this message by anyone other than the addressee is not authorized. If you are not the intended recipient, or an agent of the intended recipient, any disclosure, copying, or distribution of the message or any action or omission taken by you in reliance on it, is prohibited and may be unlawful. If you have received this message in error, please contact the sender immediately and permanently delete the original e-mail, attachment(s), and any copies. ============================================================================== From bill501 <@t> mindspring.com Wed Nov 14 15:44:49 2007 From: bill501 <@t> mindspring.com (Bill) Date: Wed Nov 14 15:45:06 2007 Subject: [Histonet] Giemsa Stain In-Reply-To: <5AEC610C1CE02945BD63A395BA763EDE011B6FA5@NVCIEXCH02.NVCI.org> References: <5AEC610C1CE02945BD63A395BA763EDE011B6FA5@NVCIEXCH02.NVCI.org> Message-ID: At 9:53 AM -0800 11/14/07, Tarango, Mark wrote: >Seriously, if you're looking for H. Pylori, you should have an H. Pylori >control. I use a colon control to be sure bacteria stain. I know what H. pylori look like so do not need to see them specifically every time. I like Alcian yellow better, but we have trouble getting it. Anyone know of a good, reliable source in the US? I think IHC is overkill and too expensive. We should all hold down medical costs. Besides, most of the docs treat for H. pylori if they see an appropriate gastritis during gastroscopy whether I report H. pylori or not. Bill From dpconsult <@t> earthlink.net Wed Nov 14 15:59:35 2007 From: dpconsult <@t> earthlink.net (Dick Paulson [Source Medical Products]) Date: Wed Nov 14 15:59:59 2007 Subject: [Histonet] Animal IHC - BVDv Ab source In-Reply-To: <00bf01c826dd$2565b6f0$a1065486@auxs.umn.edu> References: <00bf01c826dd$2565b6f0$a1065486@auxs.umn.edu> Message-ID: <005801c82709$a55e0390$a3013b0a@PathoNews> Jan, You can contact: Diane Bell University of Nebraska-Vet Diagnostic Lab Fair Street & East Campus Loop Room # 113 - Histology Lab Lincoln, NE 68588 Phone : (402) 472-4634 EMail : dbell3@unl.edu Diane runs the lab here and said she would be glad to help you. This is the lab of Dr. Bruce Brodersen, the veterinary pathologist with University of Nebraska in Lincoln who developed the skin test for BVD. Hope this is useful. Dick Paulson Source Medical Products -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Wednesday, November 14, 2007 10:41 AM To: histonet Subject: [Histonet] Animal IHC - BVDv Ab source Could anyone please forward me their BVD (Bovine Viral Diarrhea virus) antibody vendor name? My previous vendor no longer supplies it, and I'm not happy with my new source (nonspecific background issues). I'd appreciate any information you may be able to provide. I need to find a new vendor as soon as possible. Thanks much. Jan Shivers UMN Vet Diag Lab St. Paul, MN shive003@umn.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Wed Nov 14 16:54:03 2007 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Wed Nov 14 16:54:14 2007 Subject: [Histonet] Colored OCT Message-ID: <57BE698966D5C54EAE8612E8941D768301FD272B@EXCHANGE3.huntingtonhospital.com> Where can I find OCT in different colors? I've seen it somewhere, but now I can't find the vendor/manufacturer. Laurie Colbert From Jason.Wiese <@t> va.gov Wed Nov 14 17:00:02 2007 From: Jason.Wiese <@t> va.gov (Wiese, Jason VHAROS) Date: Wed Nov 14 17:00:13 2007 Subject: [Histonet] remove me from list please... :) In-Reply-To: <8C9F4E811B87D5F-17E8-97DC@FWM-D32.sysops.aol.com> References: <8C9F4E811B87D5F-17E8-97DC@FWM-D32.sysops.aol.com> Message-ID: <70EEF3D43B3C164C94037D811B2BE19301D178EF@VHAV20MSGA3.v20.med.va.gov> I will be out of the office for a month or two. Please remove me from the list, and I will request to be added again when I return to business as usual. Thank You! Jason Jason Wiese, BS,CJ,CSI,HT(ASCP),PA(ASCP) From vazquezr <@t> ohsu.edu Wed Nov 14 17:10:39 2007 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Wed Nov 14 17:11:10 2007 Subject: [Histonet] Colored OCT Message-ID: We put food coloring in ours and mix, cheaper that way. Robyn From raj <@t> bluemarble.net Wed Nov 14 17:22:44 2007 From: raj <@t> bluemarble.net (Rebecca Johnson) Date: Wed Nov 14 17:22:59 2007 Subject: [Histonet] Special Stainers Message-ID: <00d901c82715$42e4da70$7b48f9d8@CHURCH> I need to know what everyone is using for Special Stainers? I now have Ventana special stainer. Have had the same machine for several years with maintance(cleaning) done on it daily, monthly and PM. Just not real happy with the service and inconsistent staining with the silver stains. Thanks for all your info. Becky From jmahoney <@t> alegent.org Wed Nov 14 17:36:08 2007 From: jmahoney <@t> alegent.org (Janice Mahoney) Date: Wed Nov 14 17:36:20 2007 Subject: [Histonet] Special Stainers In-Reply-To: <00d901c82715$42e4da70$7b48f9d8@CHURCH> References: <00d901c82715$42e4da70$7b48f9d8@CHURCH> Message-ID: <473B32080200003C00020D6F@gwia.alegent.org> I have had problems with pneumocystis staining consistency. We have tried all the recommended things, like bringing the Ag to room temp, assuring the instrument is balanced, routine decontamination, to no avail. Any suggestions are appreciated. Does anyone know of an easy , cheap, reliable AG kit for pneumo? Thanks for the help. Jan Omaha, NE "Rebecca Johnson" 11/14/2007 5:22 PM >>> I need to know what everyone is using for Special Stainers? I now have Ventana special stainer. Have had the same machine for several years with maintance(cleaning) done on it daily, monthly and PM. Just not real happy with the service and inconsistent staining with the silver stains. Thanks for all your info. Becky From mickie25 <@t> netzero.net Wed Nov 14 19:09:53 2007 From: mickie25 <@t> netzero.net (Mickie Johnson) Date: Wed Nov 14 19:10:20 2007 Subject: [Histonet] Colored OCT In-Reply-To: <57BE698966D5C54EAE8612E8941D768301FD272B@EXCHANGE3.huntingtonhospital.com> References: <57BE698966D5C54EAE8612E8941D768301FD272B@EXCHANGE3.huntingtonhospital.com> Message-ID: Dear Laurie The OCT made by Sakura does not come in colors, but a drop of food coloring and mixing by rotating the bottle end over end will do the trick. If you want darker color, use two drops. There are other brands of FS embedding media out there which do come in colors, but I like OCT the best. Mickie Mickie Johnson, B.S., HTL(ASCP) Mohs Histology Consulting Services, LLC & Mohs Lab Staffing 2507 S. Manito Blvd. Spokane, WA 99203 509-954-7134 Web: www.mohshistotemp.com & www.mohslabstaffing.com Email: mickie25@netzero.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie Colbert Sent: Wednesday, November 14, 2007 2:54 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Colored OCT Where can I find OCT in different colors? I've seen it somewhere, but now I can't find the vendor/manufacturer. Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From burch007 <@t> mc.duke.edu Wed Nov 14 21:12:41 2007 From: burch007 <@t> mc.duke.edu (James L Burchette) Date: Wed Nov 14 21:15:40 2007 Subject: [Histonet] Colored OCT In-Reply-To: <57BE698966D5C54EAE8612E8941D768301FD272B@EXCHANGE3.huntingtonhospital.com> References: <57BE698966D5C54EAE8612E8941D768301FD272B@EXCHANGE3.huntingtonhospital.com> Message-ID: Buy clear OCT. add a couple of drops of food coloring, mix. have tinted OCT. Jim Burchette -----histonet-bounces@lists.utsouthwestern.edu wrote: ----- To: waht.swest.nhs.uk Thu Nov 15 02:17:26 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu Nov 15 02:17:34 2007 Subject: [Histonet] Giemsa Stain Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F181@wahtntex2.waht.swest.nhs.uk> Maybe it's the faecal content that's prohibiting staining; too much biological matter. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From godsgalnow <@t> aol.com Thu Nov 15 06:49:32 2007 From: godsgalnow <@t> aol.com (godsgalnow@aol.com) Date: Thu Nov 15 06:49:43 2007 Subject: [Histonet] ki-67, 53, and p27 Message-ID: <8C9F59D1A6A4964-CB4-6A22@MBLK-M38.sysops.aol.com> I need some diplomatic advice...... We are doing Ki-67, p53, and p27, which are all nuclear stains.? Here is the problem......my pathologists is telling me that they should all look alike.? I asked him why and he said because they are nuclear stains.? He put the Ki-67 on the scope and told me that is what I should be shooting for in all nuclear stains.? I use breast cancer for a positive control on all 3 of these antibodies.? ANd he is telling me to titer the p27 and p53 out further because there is some faint staining on the normal epithelium in the ductal lobes.? I asked him if it was possible that they should be there and he thinks not. HELP! Roxanne ________________________________________________________________________ Email and AIM finally together. You've gotta check out free AOL Mail! - http://mail.aol.com From mohs76009 <@t> yahoo.com Thu Nov 15 07:41:24 2007 From: mohs76009 <@t> yahoo.com (Matt Bancroft) Date: Thu Nov 15 07:41:29 2007 Subject: [Histonet] Colored OCT In-Reply-To: Message-ID: <130977.43220.qm@web63409.mail.re1.yahoo.com> I use Neg 50 by Richard Allen Mickie Johnson wrote: Dear Laurie The OCT made by Sakura does not come in colors, but a drop of food coloring and mixing by rotating the bottle end over end will do the trick. If you want darker color, use two drops. There are other brands of FS embedding media out there which do come in colors, but I like OCT the best. Mickie Mickie Johnson, B.S., HTL(ASCP) Mohs Histology Consulting Services, LLC & Mohs Lab Staffing 2507 S. Manito Blvd. Spokane, WA 99203 509-954-7134 Web: www.mohshistotemp.com & www.mohslabstaffing.com Email: mickie25@netzero.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie Colbert Sent: Wednesday, November 14, 2007 2:54 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Colored OCT Where can I find OCT in different colors? I've seen it somewhere, but now I can't find the vendor/manufacturer. Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. From mohs76009 <@t> yahoo.com Thu Nov 15 07:45:16 2007 From: mohs76009 <@t> yahoo.com (Matt Bancroft) Date: Thu Nov 15 07:45:20 2007 Subject: [Histonet] Registry Exam Materials In-Reply-To: <90354A475B420441B2A0396E5008D4965E1FFE@copc-sbs.COPC.local> Message-ID: <276571.87435.qm@web63414.mail.re1.yahoo.com> Freda Carson book: HISTOTECHNOLOGY A SELF-INSTRUCTIONAL TEXT--second edition This is the only book that I studied from Thomas Jasper wrote: Hello Histofolk, I've got a temp working here preparing for the registry exam. Any recommendations for study materials? Preferably something that's inexpensive and easily obtained. Thanks, Tom J. Thomas Jasper HT (ASCP) BAS Histology Supervisor Central Oregon Regional Pathology Services Bend, Oregon 97701 tjasper@copc.net 541/693-2677 ext. 4050 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From vazquezr <@t> ohsu.edu Thu Nov 15 08:28:46 2007 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Nov 15 08:29:18 2007 Subject: [Histonet] Colored OCT Message-ID: Mickey, We like lots of color so we use 4-6 drops, depending on the color we want to achieve. BRIGHT COLORS!! And the variety of colors! It is amazing the little things that make us happy!!! Robyn OHSU From TMcNemar <@t> lmhealth.org Thu Nov 15 08:56:23 2007 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Thu Nov 15 08:56:14 2007 Subject: [Histonet] Colored OCT In-Reply-To: <57BE698966D5C54EAE8612E8941D768301FD272B@EXCHANGE3.huntingtonhospital.com> Message-ID: <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4F5@lmhsmail.lmhealth.org> So, in answer to your question, you can get it from BBC Biochemical (www.bbcus.com) We tried it but didn't like it. When frozen, there wasn't a lot of color difference. We have since gone with adding food coloring to the Tissue Tek mounting media. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Laurie Colbert Sent: Wednesday, November 14, 2007 5:54 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Colored OCT Where can I find OCT in different colors? I've seen it somewhere, but now I can't find the vendor/manufacturer. Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Dorothy.L.Webb <@t> HealthPartners.Com Thu Nov 15 09:07:57 2007 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Thu Nov 15 09:08:14 2007 Subject: [Histonet] Pin4 In-Reply-To: <01MNPJ4NKODE000Y2Y@Dino.HealthPartners.Com> Message-ID: <0E394B648E5284478A6CCB78E5AFDA2705635289@hpes1.HealthPartners.int> Cell marque also carries the cocktail!!! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, November 14, 2007 7:58 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 48, Issue 18 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: Antibody cocktails (godsgalnow@aol.com) 2. Histology equipments (Ms. K?tia Cristina Catunda) 3. Re: staining of negative controls with IHC (Johnson, Teri) 4. processing (Webb, Dorothy L) 5. RE: processing (Tarango, Mark) 6. RE: processing (Tom McNemar) 7. Histonet down? (Thomas Jasper) 8. Re: Histology equipments (Rene J Buesa) 9. RE: Antibody cocktails (Barry Madigan) 10. microtomy technical question (Eva C Andersson) 11. Re: microtomy technical question (Rene J Buesa) 12. It may be more than just a microtomy technical question (Gayle Callis) 13. Giemsa Stain (Schaundra Walton) 14. RE: automated paraffin embedding systems (Bartlett, Jeanine (CDC/CCID/NCZVED)) 15. RE: automated paraffin embedding systems (Bartlett, Jeanine (CDC/CCID/NCZVED)) 16. Re: Giemsa Stain (Rene J Buesa) 17. RE: Giemsa Stain (Marshall Terry Dr, Consultant Histopathologist) 18. RE: Giemsa Stain (Rene J Buesa) 19. RE: Giemsa Stain (Kemlo Rogerson) 20. Re: Giemsa Stain (koellingr@comcast.net) 21. RE: Giemsa Stain (Marshall Terry Dr, Consultant Histopathologist) 22. RE: Giemsa Stain (Marshall Terry Dr, Consultant Histopathologist) 23. RE: Giemsa Stain (Kemlo Rogerson) 24. RE: Giemsa Stain (Rene J Buesa) ---------------------------------------------------------------------- Message: 1 Date: Tue, 13 Nov 2007 13:06:20 -0500 From: godsgalnow@aol.com Subject: Re: [Histonet] Antibody cocktails To: CBraaten@Cheshire-Med.COM, histonet@lists.utsouthwestern.edu Message-ID: <8C9F437075F6496-EC4-5B3F@WEBMAIL-MB20.sysops.aol.com> Content-Type: text/plain; charset="us-ascii" Christine, My understanding is that other vendors do have something similar to the cocktail, but they only contain 2 of the antibodies and the third has to be added at another step, not completely sure on that.? But I do use the BioCare PIN-4 cocktail and have been doing so for 3 years now with no complaints; I?have done?a few thousands of these so far.? I use their Mach 2 Double Stain detection system with the cocktail and it works like a beaut. Roxanne Soto HT(ASCP)QIHC -----Original Message----- From: Christine I. Braaten To: histonet@lists.utsouthwestern.edu Sent: Tue, 13 Nov 2007 12:32 pm Subject: [Histonet] Antibody cocktails Can anybody tell me if there is another company besides Biocare that offers an antibody cocktail with P504S, HMW CK, and p63? Does anybody use the Biocare antibody detection system? I'd appreciate some feedback. Thanks. CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients 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 electronic mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________ Email and AIM finally together. You've gotta check out free AOL Mail! - http://mail.aol.com ------------------------------ Message: 2 Date: Tue, 13 Nov 2007 16:20:18 -0300 From: Ms. K?tia Cristina Catunda Subject: [Histonet] Histology equipments To: "'Histology Network'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi there, Beside Sakura, Therm and Mopec, does anybody has anyother manufacturers for histology equipments for recommendation (embedding centers, grossing tables, cassette printers, slide stainers, etc) Tks a lot Ms. K?tia Catunda Produ??o +55 12 3203-0612 (direto) +55 12 3203-0633 (PABX) www.cipax.com.br katia.catunda@cipax.com.br ------------------------------ Message: 3 Date: Tue, 13 Nov 2007 12:31:34 -0600 From: "Johnson, Teri" Subject: [Histonet] Re: staining of negative controls with IHC To: Message-ID: Content-Type: text/plain; charset="US-ASCII" MaryAnn Dixon wrote: "We are currently retrieving animal tissue in a pressure cooker with a program of 125 degrees for 30 secs. This protocol was taken from biocare but I'm sure this is for human tissue. Most of the tissue is retrieved with Biocare's Reveal (pH 6.0). We are receiving some background staining of our negative controls whereas if we did not retrieve them, there is none. We were staining for KI-67 which is a nuclear stain. I am using a tonsil for the control. The negative control showed some sporatic cytoplasmic staining on the Universal negative control from biocare as well as deionized water. Since then we have taken the temperature down to 115 degrees and increased the time to 25 minutes. It seems to have decreased the cytoplasmic staining but there is still some lingering. Anyone out there have any ideas??? I'm sure there is a better protocol for retrieving animal tissues. It has to be a time and temperature thing!" MaryAnn, you don't mention what type of animal you are staining, and what your IHC protocol looks like. I would never use a universal negative control when doing immunostaining on animals. Use only the Igs from the species the primary antibody was made in. If you are using a monoclonal mouse KI-67, you would need to use non-immune mouse IgGs of the same isotype of your primary antibody. If your antibody is a rabbit polyclonal or monoclonal, you'd need to use non-immune rabbit IgG. It's also important that you match the Ig concentration fo the non-immune (negative) control to the Ig concentration of your primary antibody. If you are using pre-diluted antibodies, you may be using different Ig concentrations. Try an additional run to see if your detection system is causing the staining. Run a tonsil, and do the pressure cooker retrieval as usual, however and omit the primary antibody, using only diluent or buffer in the incubation. Then continue your IHC protocol as usual. If there is still some background/non-specific staining, it's due to your detection system (kit?) and not to the antibody or negative control reagent. Additionally, I never use a universal reagent for detection. Making sure your antibody reactions are appropriate are difficult enough without throwing in additional animal species you don't need. If you still have specific questions, don't hesitate to contact me off-list and I'll be happy to take a look at your protocol and make some recommendations. Good luck! Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 ------------------------------ Message: 4 Date: Tue, 13 Nov 2007 12:35:36 -0600 From: "Webb, Dorothy L" Subject: [Histonet] processing To: histonet@lists.utsouthwestern.edu Message-ID: <0E394B648E5284478A6CCB78E5AFDA2705635279@hpes1.HealthPartners.int> Content-Type: text/plain; charset="us-ascii" Now that we can no longer use alcoholic fixatives on breast tissue, is there anyone out there that is putting a slight temperature on the reagents on the processor to enhance fixation and dehydration? I have thought of using stronger alcohol grades and an extra xylene, which would help in the dehydration of the lipids. This would be a dedeicated processor for fatty tissue types only. What is everyone's thoughts and thanks for all input! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. ------------------------------ Message: 5 Date: Tue, 13 Nov 2007 10:42:51 -0800 From: "Tarango, Mark" Subject: RE: [Histonet] processing To: "Webb, Dorothy L" , histonet@lists.utsouthwestern.edu Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F9C@NVCIEXCH02.NVCI.org> Content-Type: text/plain; charset=us-ascii Can you use 20% formalin on breast tissue? Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Webb, Dorothy L Sent: Tuesday, November 13, 2007 10:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing Now that we can no longer use alcoholic fixatives on breast tissue, is there anyone out there that is putting a slight temperature on the reagents on the processor to enhance fixation and dehydration? I have thought of using stronger alcohol grades and an extra xylene, which would help in the dehydration of the lipids. This would be a dedeicated processor for fatty tissue types only. What is everyone's thoughts and thanks for all input! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== ------------------------------ Message: 6 Date: Tue, 13 Nov 2007 14:03:15 -0500 From: "Tom McNemar" Subject: RE: [Histonet] processing To: "Webb, Dorothy L" , Message-ID: <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4F4@lmhsmail.lmhealth.org> Content-Type: text/plain; charset="iso-8859-1" Why can't you use alcoholic fixatives? Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Webb, Dorothy L Sent: Tuesday, November 13, 2007 1:36 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing Now that we can no longer use alcoholic fixatives on breast tissue, is there anyone out there that is putting a slight temperature on the reagents on the processor to enhance fixation and dehydration? I have thought of using stronger alcohol grades and an extra xylene, which would help in the dehydration of the lipids. This would be a dedeicated processor for fatty tissue types only. What is everyone's thoughts and thanks for all input! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Tue, 13 Nov 2007 08:13:28 -0800 From: "Thomas Jasper" Subject: [Histonet] Histonet down? To: Message-ID: <90354A475B420441B2A0396E5008D4965E1FF9@copc-sbs.COPC.local> Content-Type: text/plain; charset="us-ascii" Anyone having trouble with the Histonet? Haven't seen any postings for a while. T. Jasper ------------------------------ Message: 8 Date: Tue, 13 Nov 2007 12:40:13 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Histology equipments To: "Ms. K?tia Cristina Catunda" , 'Histology Network' Message-ID: <9152.47378.qm@web61212.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 There are many, but Sakura is the best. Ren? J. "Ms. K?tia Cristina Catunda" wrote: Hi there, Beside Sakura, Therm and Mopec, does anybody has anyother manufacturers for histology equipments for recommendation (embedding centers, grossing tables, cassette printers, slide stainers, etc) Tks a lot Ms. K?tia Catunda Produ??o +55 12 3203-0612 (direto) +55 12 3203-0633 (PABX) www.cipax.com.br katia.catunda@cipax.com.br _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. ------------------------------ Message: 9 Date: Wed, 14 Nov 2007 06:58:44 +1000 From: "Barry Madigan" Subject: RE: [Histonet] Antibody cocktails To: "'Christine I. Braaten'" , Message-ID: <000001c82638$012fe800$038fb800$@com.au> Content-Type: text/plain; charset="iso-8859-1" Hi Christine, We make up our own antibody cocktail of 34?E12 (DAKO dilution 1:150) and P63 (DAKO 1:400) and stain the section with this cocktail with a Polymer peroxidise detection system (DAB).(Heat retrieval in a high pH retrieval solution for 20 min at 100 C. The stained slides are washed and darkened using a copper sulphate solution. Placed in a buffer wash and then stained for the second time with P504S (AMACR/Racemase) at a dilution of 1:150 with a Polymer Alkaline Phosphatase detection system (Fast Red). We do this all on the BondMax Immunostainer. We have been doing this for over 6 months now and did it this way because we already had all the concentrated antibodies. Proper fixation is vital. Hope this helps. Regards Barry B.Madigan Pathology Queensland Royal Brisbane Campus Australia ------------------------------ Message: 10 Date: Tue, 13 Nov 2007 16:32:03 -0500 From: Eva C Andersson Subject: [Histonet] microtomy technical question To: histonet@lists.utsouthwestern.edu Message-ID: <163fae165e00.165e00163fae@imap.georgetown.edu> Content-Type: text/plain; charset=us-ascii Hello everyone, I have some questions regarding microtomy. We place our blocks face down on ice. The problem is that some tissues take a very long time before they are ready to be cut(some more than 6 hours). For some tissues like kidney samples (mouse tissue) we have been using glycerol on the ice. This does seem to cut down on the time needed. My question is which other tissues can I use this technique on? Do you have any other suggestions for how to get the tissue hydrated for cutting? Thank you for your help, Eva Permaul Georgetown University ------------------------------ Message: 11 Date: Tue, 13 Nov 2007 13:39:58 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] microtomy technical question To: Eva C Andersson , histonet@lists.utsouthwestern.edu Message-ID: <975592.48450.qm@web61220.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Eva: This type of problem (requiring ice, glycerol, and others) to allow sectioning usually stem from either a fixing or processing defficiency. I suggest you to check your fixing times and processing protocols or you will keep having this type of difficulties. Tissues (any type) properly infiltrated (after adequate fixation/processing) do not need anything but a light cooling to allow ribbons to be taken. Ren? J. Eva C Andersson wrote: Hello everyone, I have some questions regarding microtomy. We place our blocks face down on ice. The problem is that some tissues take a very long time before they are ready to be cut(some more than 6 hours). For some tissues like kidney samples (mouse tissue) we have been using glycerol on the ice. This does seem to cut down on the time needed. My question is which other tissues can I use this technique on? Do you have any other suggestions for how to get the tissue hydrated for cutting? Thank you for your help, Eva Permaul Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. ------------------------------ Message: 12 Date: Tue, 13 Nov 2007 14:57:37 -0700 From: "Gayle Callis" Subject: [Histonet] It may be more than just a microtomy technical question To: "Eva C Andersson" , "Histonet" Message-ID: <000701c82640$33f907a0$6501a8c0@DHXTS541> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Eva, It sounds as though you are processing your tissues too long, and too much water is being removed (over dehydration.) In general, a properly processed mouse tissue after a short schedule will need a bit of a water soak, but not for 6 hours. Another trick is try room temperature or even warm water for a few minutes, then go to ice water, and be careful to NOT cut away what you have soaked - be careful to reapproach the blade in order to cut the sections. Some people have suggested floating blocks face down on a waterbath for a very short time. if you post your processing schedule, Histonetters can help solve some of this problem. Gayle Callis HT,HTL,MT(ASCP) ----- Original Message ----- From: "Eva C Andersson" To: Sent: Tuesday, November 13, 2007 2:32 PM Subject: [Histonet] microtomy technical question > Hello everyone, > I have some questions regarding microtomy. > We place our blocks face down on ice. The problem is that some tissues > take a very long time before they are ready to be cut(some more than 6 > hours). For some tissues like kidney samples (mouse tissue) we have been > using glycerol on the ice. This does seem to cut down on the time needed. > My question is which other tissues can I use this technique on? Do you > have any other suggestions for how to get the tissue hydrated for cutting? > Thank you for your help, > Eva Permaul > Georgetown University > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Tue, 13 Nov 2007 15:00:06 -0800 (PST) From: Schaundra Walton Subject: [Histonet] Giemsa Stain To: Histonet Message-ID: <625759.21602.qm@web58904.mail.re1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. ------------------------------ Message: 14 Date: Wed, 14 Nov 2007 05:48:41 -0500 From: "Bartlett, Jeanine (CDC/CCID/NCZVED)" Subject: RE: [Histonet] automated paraffin embedding systems To: "Rene J Buesa" , "Lesley Bechtold" , "Histology Network" Message-ID: <34BB307EFC9A65429BBB49E330675F72045E21F1@LTA3VS003.ees.hhs.gov> Content-Type: text/plain; charset=iso-8859-1 Slight correction. The Xpress microwave tissue processor does not require a special type of cassette. The AutoTEC does, and it can be used on the Xpress or in a traditional processor. But the Xpress can use the same type of plastic cassette used in any traditional processor. Jeanine Bartlett, BS, HT(ASCP)QIHC Centers for Disease Control and Prevention Infectious Diseases Pathology Branch 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, November 13, 2007 12:04 PM To: Lesley Bechtold; Histology Network Subject: Re: [Histonet] automated paraffin embedding systems This embedding instrument is manufactured by Sakura. It is not that the instrument orients the tissue, that is done when the tissues are placed in the cassette and held in place by a plastic frame that will be sectioned along with the tissue when the block is prepared. To function ideally it requires a special type of cassette and cassette holder (that is used alsao by the Sakura Xpress microwave tissue processor). Ren? J. Lesley Bechtold wrote: Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Wed, 14 Nov 2007 05:50:24 -0500 From: "Bartlett, Jeanine (CDC/CCID/NCZVED)" Subject: RE: [Histonet] automated paraffin embedding systems To: "Mike Pence" , "Rene J Buesa" , "Lesley Bechtold" , "Histology Network" Message-ID: <34BB307EFC9A65429BBB49E330675F72045E21F2@LTA3VS003.ees.hhs.gov> Content-Type: text/plain; charset=iso-8859-1 I use both machines. You can use the AutoTEC cassette on any type of processor (including the Xpress) and you do not have to use the AutoTEC and the Xpress both. They can be used independently. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mike Pence Sent: Tuesday, November 13, 2007 12:10 PM To: Rene J Buesa; Lesley Bechtold; Histology Network Subject: RE: [Histonet] automated paraffin embedding systems Is anyone out there using this function of the Sakura Xpress? Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, November 13, 2007 11:04 AM To: Lesley Bechtold; Histology Network Subject: Re: [Histonet] automated paraffin embedding systems This embedding instrument is manufactured by Sakura. It is not that the instrument orients the tissue, that is done when the tissues are placed in the cassette and held in place by a plastic frame that will be sectioned along with the tissue when the block is prepared. To function ideally it requires a special type of cassette and cassette holder (that is used alsao by the Sakura Xpress microwave tissue processor). Ren? J. Lesley Bechtold wrote: Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Wed, 14 Nov 2007 05:01:09 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Giemsa Stain To: Schaundra Walton , Histonet Message-ID: <387310.3350.qm@web61217.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Giemsa, as any other stain, is just that, a stain, and stains do not (cannot) have "internal controls". As with any other HC procedure you have to use a positive control along with your tissue. To completely demonstrate that your Giemsa procedure worked as expected, you coul use a blood smear or an appendix section as controls. That will depend on your pathologist's whims. Ren? J. Schaundra Walton wrote: We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. ------------------------------ Message: 17 Date: Wed, 14 Nov 2007 13:22:36 -0000 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Giemsa Stain To: "Rene J Buesa" , "Schaundra Walton" , "Histonet" Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE79@TRFT-EX01.xRothGen.nhs.uk> Content-Type: text/plain; charset="iso-8859-1" Sorry Rene, on this occasion I can't agree. Giemsa is not on this occasion being used to bring out granules as it would in it's normal use as a blood/marrow stain. It is just used as a simple quick stain, which will pick up Helicobacter inter alia. If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: 14 November 2007 13:01 To: Schaundra Walton; Histonet Subject: Re: [Histonet] Giemsa Stain Giemsa, as any other stain, is just that, a stain, and stains do not (cannot) have "internal controls". As with any other HC procedure you have to use a positive control along with your tissue. To completely demonstrate that your Giemsa procedure worked as expected, you coul use a blood smear or an appendix section as controls. That will depend on your pathologist's whims. Ren? J. Schaundra Walton wrote: We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Wed, 14 Nov 2007 05:41:09 -0800 (PST) From: Rene J Buesa Subject: RE: [Histonet] Giemsa Stain To: "Marshall Terry Dr, Consultant Histopathologist" , Schaundra Walton , Histonet Message-ID: <246072.35484.qm@web61218.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I was referring at the fact that IF some coloration took place (of any kind) the procedure worked, and there was a staining. I was not referring to stain specificity. Ren? J. "Marshall Terry Dr, Consultant Histopathologist" wrote: Sorry Rene, on this occasion I can't agree. Giemsa is not on this occasion being used to bring out granules as it would in it's normal use as a blood/marrow stain. It is just used as a simple quick stain, which will pick up Helicobacter inter alia. If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: 14 November 2007 13:01 To: Schaundra Walton; Histonet Subject: Re: [Histonet] Giemsa Stain Giemsa, as any other stain, is just that, a stain, and stains do not (cannot) have "internal controls". As with any other HC procedure you have to use a positive control along with your tissue. To completely demonstrate that your Giemsa procedure worked as expected, you coul use a blood smear or an appendix section as controls. That will depend on your pathologist's whims. Ren? J. Schaundra Walton wrote: We've been using a Giemsa stain for our H. pylori cases without a control tissue. It has come to my attention that this may not be good practice. I was under the impression that the Giemsa stain had an internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. ------------------------------ Message: 19 Date: Wed, 14 Nov 2007 13:42:27 -0000 From: "Kemlo Rogerson" Subject: RE: [Histonet] Giemsa Stain To: "Marshall Terry Dr, Consultant Histopathologist" , "Rene J Buesa" , "Schaundra Walton" , "Histonet" Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F171@wahtntex2.waht.swest.nhs.uk> Content-Type: text/plain; charset="us-ascii" "If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose." Terry If it's not blue then it hasn't worked or the organism isn't there! Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation ------------------------------ Message: 20 Date: Wed, 14 Nov 2007 13:43:25 +0000 From: koellingr@comcast.net Subject: Re: [Histonet] Giemsa Stain To: Rene J Buesa , Schaundra Walton , Histonet Message-ID: <111420071343.9538.473AFB7D00086C1A0000254222058863609D09020704040A0105@comcast.net> Content-Type: text/plain Rene, Are you saying a histochemical stain for an organism (Helicobactor, AFB, spirochetes, etc) must have a positive control with which I'd agree? But are you saying that for "any" HC stain like elastic, trichrome, PAS, alcian blue, etc that internal controls don't (cannot) even exist? Thanks, Ray Koelling PhenoPath Labs Seattle, WA -------------- Original message -------------- From: Rene J Buesa > Giemsa, as any other stain, is just that, a stain, and stains do not > (cannot) > have "internal controls". > As with any other HC procedure you have to use a positive control along with > your tissue. > To completely demonstrate that your Giemsa procedure worked as expected, you > coul use a blood smear or an appendix section as controls. That will depend on > your pathologist's whims. > Ren$B!&(BJ. > > Schaundra Walton wrote: > We've been using a Giemsa stain for our H. pylori cases without a control > tissue. It has come to my attention that this may not be good practice. I was > under the impression that the Giemsa stain had an internal control. Is anyone > else using a this stain for H. pylori? Are you running a tissue control with it? > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: Wed, 14 Nov 2007 13:50:33 -0000 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Giemsa Stain To: "Kemlo Rogerson" , "Rene J Buesa" , "Schaundra Walton" , "Histonet" Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE7A@TRFT-EX01.xRothGen.nhs.uk> Content-Type: text/plain; charset="us-ascii" No. The tissue can still be blue even if the organism isn't there. BTW, has anyone ever seen a Giemsa that doesn't stain? I can't really imagine it. Terry -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@waht.swest.nhs.uk] Sent: 14 November 2007 13:42 To: Marshall Terry Dr, Consultant Histopathologist; Rene J Buesa; Schaundra Walton; Histonet Subject: RE: [Histonet] Giemsa Stain "If the tissue is blue, it has worked, at least to the extent it needs to have done for this purpose." Terry If it's not blue then it hasn't worked or the organism isn't there! Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation ------------------------------ Message: 22 Date: Wed, 14 Nov 2007 13:53:15 -0000 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Giemsa Stain To: , "Rene J Buesa" , "Schaundra Walton" , "Histonet" Message-ID: <5C0BED61F529364E86309CADEA63FEF2F3AE7B@TRFT-EX01.xRothGen.nhs.uk> Content-Type: text/plain; charset="us-ascii" So we are singing from the same hymn sheet.... Stains can't have internal controls. Tissues can, in relation to some stains. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of koellingr@comcast.net Sent: 14 November 2007 13:43 To: Rene J Buesa; Schaundra Walton; Histonet Subject: Re: [Histonet] Giemsa Stain Rene, Are you saying a histochemical stain for an organism (Helicobactor, AFB, spirochetes, etc) must have a positive control with which I'd agree? But are you saying that for "any" HC stain like elastic, trichrome, PAS, alcian blue, etc that internal controls don't (cannot) even exist? Thanks, Ray Koelling PhenoPath Labs Seattle, WA -------------- Original message -------------- From: Rene J Buesa > Giemsa, as any other stain, is just that, a stain, and stains do not > (cannot) have "internal controls". > As with any other HC procedure you have to use a positive control > along with your tissue. > To completely demonstrate that your Giemsa procedure worked as > expected, you coul use a blood smear or an appendix section as > controls. That will depend on your pathologist's whims. > Ren$B!&(BJ. > > Schaundra Walton wrote: > We've been using a Giemsa stain for our H. pylori cases without a > control tissue. It has come to my attention that this may not be good > practice. I was under the impression that the Giemsa stain had an > internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@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: 23 Date: Wed, 14 Nov 2007 13:54:07 -0000 From: "Kemlo Rogerson" Subject: RE: [Histonet] Giemsa Stain To: "Marshall Terry Dr, Consultant Histopathologist" , "Rene J Buesa" , "Schaundra Walton" , "Histonet" Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F172@wahtntex2.waht.swest.nhs.uk> Content-Type: text/plain; charset="us-ascii" "No. The tissue can still be blue even if the organism isn't there." Terry I know what you mean; I get days like that too. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation ------------------------------ Message: 24 Date: Wed, 14 Nov 2007 05:57:03 -0800 (PST) From: Rene J Buesa Subject: RE: [Histonet] Giemsa Stain To: "Marshall Terry Dr, Consultant Histopathologist" , koellingr@comcast.net, Schaundra Walton , Histonet Message-ID: <588286.70996.qm@web61222.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Amen! Ren? J. "Marshall Terry Dr, Consultant Histopathologist" wrote: So we are singing from the same hymn sheet.... Stains can't have internal controls. Tissues can, in relation to some stains. Terry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of koellingr@comcast.net Sent: 14 November 2007 13:43 To: Rene J Buesa; Schaundra Walton; Histonet Subject: Re: [Histonet] Giemsa Stain Rene, Are you saying a histochemical stain for an organism (Helicobactor, AFB, spirochetes, etc) must have a positive control with which I'd agree? But are you saying that for "any" HC stain like elastic, trichrome, PAS, alcian blue, etc that internal controls don't (cannot) even exist? Thanks, Ray Koelling PhenoPath Labs Seattle, WA -------------- Original message -------------- From: Rene J Buesa > Giemsa, as any other stain, is just that, a stain, and stains do not > (cannot) have "internal controls". > As with any other HC procedure you have to use a positive control > along with your tissue. > To completely demonstrate that your Giemsa procedure worked as > expected, you coul use a blood smear or an appendix section as > controls. That will depend on your pathologist's whims. > Ren$B!&(BJ. > > Schaundra Walton wrote: > We've been using a Giemsa stain for our H. pylori cases without a > control tissue. It has come to my attention that this may not be good > practice. I was under the impression that the Giemsa stain had an > internal control. Is anyone else using a this stain for H. pylori? Are you running a tissue control with it? > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 18 **************************************** ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From gu.lang <@t> gmx.at Thu Nov 15 09:12:16 2007 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Thu Nov 15 09:12:24 2007 Subject: WG: [Histonet] MyoD1 Message-ID: <001601c82799$e94a2850$6412a8c0@dielangs.at> 1:30, cc1 32 min, 32 min ink., 37?C Gudrun Lang Biomed. Analytikerin Histolabor Akh Linz Krankenhausstr. 9 4020 Linz +43(0)732/7806-6754 -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von San Tin Gesendet: Dienstag, 13. November 2007 02:35 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] MyoD1 Dear All, Can anyone give me the protocol of MyoD1 clone 5.8A on Ventana IHC autostainer? I have problem with staining. Thanks San _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jengirl1014 <@t> yahoo.com Thu Nov 15 09:19:13 2007 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Thu Nov 15 09:19:20 2007 Subject: [Histonet] tissue processors Message-ID: <98496.35382.qm@web62411.mail.re1.yahoo.com> I was wondering if anyone had a good protocol for mouse tissue on a Shandon Excelsior Tissue Processor. Thanks Jennifer K. Sipes, ALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 410-955-9688 fax: 410-955-9677 cell: 443-631-6361 e-mail: jsipes1@jhmi.edu ____________________________________________________________________________________ Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. http://overview.mail.yahoo.com/ From ksecrest <@t> hsc.wvu.edu Thu Nov 15 09:37:02 2007 From: ksecrest <@t> hsc.wvu.edu (Kimberly Secrest) Date: Thu Nov 15 09:37:53 2007 Subject: [Histonet] Hi all Message-ID: <473C2154.C068.0078.0@hsc.wvu.edu> Does anybody have or know of anyone willing to donate histology equipment (i.e. microtomes, tissue processor, embedding center, routine stainer, etc) or supplies (i.e. slides, cassettes, reagents) for the start up of a histotechnology program? Kimberly Secrest, HTL, QIHC Instructor Department of Pathology School of Medicine West Virginia University From FMonson <@t> wcupa.edu Thu Nov 15 09:57:31 2007 From: FMonson <@t> wcupa.edu (Monson, Frederick ) Date: Thu Nov 15 09:57:41 2007 Subject: [Histonet] microtomy technical question In-Reply-To: <163fae165e00.165e00163fae@imap.georgetown.edu> References: <163fae165e00.165e00163fae@imap.georgetown.edu> Message-ID: <641CEFFC7E5B6C42AB59539653FD082304A90A58@wcu-ex-emp2.PASSHE.LCL> I never respond without an answer from outside the bag, so to speak. 1. I once was quite ill (at the age of 40) and received the news that my CBC required a bone marrow follow up. I asked if the differential was at fault and heard a 'Yes'. I asked if the differential was performed manually. The answer was 'No'. I demanded a manual differential. The bone marrow was cancelled. Don't trust machines too far is one of my life lessons. 2. I never had to use watered or iced paraffin blocks BEFORE I was required by volume to process tissues in an auto processor. Prior to using auto, I had very early on learned the consequences of over-dehydrating, over-clearing and especially overcooking my specimens and avoided doing so. Now with so many 110V AC lines measuring 128V AC or <110V AC on occasion (the latter does no harm to the tissue but lengthen the times in each bath), why is there a question about problems with the outcomes? 3. For my SEM, I have a $5,600 Uninterruptible Power Supply (UPS) that regulates the AC to within the specs of my instrument so that I am not at the mercy of PECO - our local supplier - when I run long-duration automated analyses. Most institutions are treated by the power companies as industrial customers. The consequence is that power fluctuations are much more frequent. Industry compensates by investing in regulation wherever it is required. Institutions, on the other hand, are often ignorant of how much or little power fluctuations affect performance in various organizational modules - like laboratories in which analyses are performed. One will not see the difference between 128 and 108 in the performance of the coffee maker. Overnight, no one is watching and monitoring. Hope this helps, Fred Monson Frederick C. Monson, PhD Technical Director Microanalysis and Imaging Research and Training Center (MIRTC) Large Scientific Instrument Core Geology, West Chester University S. Church St. and W. Rosedale Ave. West Chester, PA, 19320 610-738-0437 fmonson@wcupa.edu New Scheduler: http://lexspiac.wcupa.edu/cgi-bin/ureserve_gold.pl Web Page: http://lexspiac.wcupa.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eva C Andersson Sent: Tuesday, November 13, 2007 4:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] microtomy technical question Hello everyone, I have some questions regarding microtomy. We place our blocks face down on ice. The problem is that some tissues take a very long time before they are ready to be cut(some more than 6 hours). For some tissues like kidney samples (mouse tissue) we have been using glycerol on the ice. This does seem to cut down on the time needed. My question is which other tissues can I use this technique on? Do you have any other suggestions for how to get the tissue hydrated for cutting? Thank you for your help, Eva Permaul Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RSRICHMOND <@t> aol.com Thu Nov 15 10:13:12 2007 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Thu Nov 15 10:13:22 2007 Subject: [Histonet] Re: Giemsa Stain Message-ID: Bill (good ol' Bill - where?) says of staining Helicobacter: >>I like Alcian yellow better, but we have trouble getting it. Anyone know of a good, reliable source in the US? I think IHC is overkill and too expensive.<< The original chemical synthesis of Alcian yellow is not environmentally safe, though the dye may still be made in countries that are not concerned about such matters. Dick Dapson at Anatech told us some time ago that he had achieved an environmentally acceptable synthesis of Alcian yellow, but that the product did not have a satisfactory shelf life. Anatech offers a substitute for Alcian yellow, which I have not seen. (I have no connection with Anatech.) Bob Richmond Samurai Pathologist Knoxville TN From making <@t> ufl.edu Thu Nov 15 10:29:36 2007 From: making <@t> ufl.edu (MKing) Date: Thu Nov 15 10:29:56 2007 Subject: [Histonet] pepsin Message-ID: <473C73F0.3010607@ufl.edu> An antibody vendor recommended pepsin pretreatment prior to primary antibody incubation of our free-floating 40 um formaldehyde-fixed rat brain frozen sections, but provided a protocol that only listed pepsin by weight. Since enzyme activity ranges from 3 to 4500 units/mg in commercial sources, does anyone have a rough idea what kind of activity is sufficient without destroying the tissue (which is what happened with the first test). Anybody find this approach worthwhile with similar tissue? Thanks, Mike King UF Pharmacology & Therapeutics From mauger <@t> email.chop.edu Thu Nov 15 10:44:01 2007 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Thu Nov 15 10:44:12 2007 Subject: [Histonet] pepsin Message-ID: Mike, We use pepsin at 2mgs/ml for digestion of formalin fixed, paraffin embedded tissue. We use it at 37C for 10 to 30 minutes. We buy powdered pepsin from Dako #S3002. Don't know what will happen to frozen tissue. Good luck, Jo Mauger From sally.norton <@t> seattlechildrens.org Thu Nov 15 10:42:46 2007 From: sally.norton <@t> seattlechildrens.org (Norton, Sally) Date: Thu Nov 15 10:45:12 2007 Subject: [Histonet] Colored OCT In-Reply-To: References: Message-ID: I have to ask the question. For what purpose do you use colored OCT? We here, at Children's in Seattle, have never heard of it. Thank you. Sally Norton, HT Children's Hospital and Regional Medical Center Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Thursday, November 15, 2007 6:29 AM To: laurie.colbert@huntingtonhospital.com; histonet@lists.utsouthwestern.edu; mickie25@netzero.net Subject: RE: [Histonet] Colored OCT Mickey, We like lots of color so we use 4-6 drops, depending on the color we want to achieve. BRIGHT COLORS!! And the variety of colors! It is amazing the little things that make us happy!!! Robyn OHSU _______________________________________________ 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 and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From Dorothy.L.Webb <@t> HealthPartners.Com Thu Nov 15 11:06:11 2007 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Thu Nov 15 11:06:18 2007 Subject: [Histonet] Colored OCT Message-ID: <0E394B648E5284478A6CCB78E5AFDA270563528D@hpes1.HealthPartners.int> TBS carries the colored OCT. The order number is H-TFM and is available in red or blue. Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From bill501 <@t> mindspring.com Thu Nov 15 11:27:14 2007 From: bill501 <@t> mindspring.com (Bill) Date: Thu Nov 15 11:27:21 2007 Subject: [Histonet] Re: Giemsa Stain In-Reply-To: References: Message-ID: At 11:13 AM -0500 11/15/07, Robert Richmond wrote: >Bill (good ol' Bill - where?) says of staining Helicobacter: I wondered why. Thanks Bob. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board/dhp/ From karenadams <@t> comcast.net Thu Nov 15 11:44:56 2007 From: karenadams <@t> comcast.net (karenadams@comcast.net) Date: Thu Nov 15 11:45:01 2007 Subject: [Histonet] Gill's III stainer question Message-ID: <111520071744.2924.473C859800022C3F00000B6C22070206539C030E0B0E020A9D0E05@comcast.net> We are presently using Gill's III manually for H & E and on the automated stainer.....I am seeing mucin lightly staining on the automated stainer, but not the manual.....any thoughts?? -- Karen Adams Supervisor Pathology Laboratories West 9303 Park West Blvd Knoxville, TN 37923 (865) 690-2111 FAX (865) 691-1623 From rjbuesa <@t> yahoo.com Thu Nov 15 12:03:43 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 15 12:03:48 2007 Subject: [Histonet] Gill's III stainer question In-Reply-To: <111520071744.2924.473C859800022C3F00000B6C22070206539C030E0B0E020A9D0E05@comcast.net> Message-ID: <799024.13950.qm@web61221.mail.yahoo.com> Probably because we tend to shake in/out quickier when staining manually that the stainer does. It takes to the stainer more time to take slides out-transport to the next station- and into the next station than we to us manually. That extra time can account for the difference. Ren? J. karenadams@comcast.net wrote: We are presently using Gill's III manually for H & E and on the automated stainer.....I am seeing mucin lightly staining on the automated stainer, but not the manual.....any thoughts?? -- Karen Adams Supervisor Pathology Laboratories West 9303 Park West Blvd Knoxville, TN 37923 (865) 690-2111 FAX (865) 691-1623 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. From SDrew <@t> uwhealth.org Thu Nov 15 12:09:35 2007 From: SDrew <@t> uwhealth.org (Drew Sally A.) Date: Thu Nov 15 12:09:39 2007 Subject: [Histonet] IgG4 antibody Message-ID: I've been given the task of trying to find out if anyone is using an IgG4 for IHC and could run a slide for someone. The pathologist brought in a post-doc. oncologist with a paper that used an antibody from Zymed. Any takers our there? Thank you! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 From amylee779 <@t> yahoo.com Thu Nov 15 12:14:24 2007 From: amylee779 <@t> yahoo.com (Amy Lee) Date: Thu Nov 15 12:14:27 2007 Subject: [Histonet] which species for stromal cell? Message-ID: <533046.17445.qm@web38004.mail.mud.yahoo.com> Hello, I have an antibody only recognize human tissue. Now I have a tumor tissue that is human tumor cell grow in mouse. I need to stain stromal cell. Can I use this antibody? Or I may ask the question this way: the human tumor cell grow in mouse, then is the stromal cell develope from human tumor or from mouse tissue? Thanks a lot! Amy --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. From TMcNemar <@t> lmhealth.org Thu Nov 15 12:32:54 2007 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Thu Nov 15 12:32:46 2007 Subject: [Histonet] Colored OCT In-Reply-To: Message-ID: <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4F6@lmhsmail.lmhealth.org> We use the different colors when we have multiple frozens. Frozen 'A' is alwasy clear, 'B' is always blue, etc. I just helps to keep them straight. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Norton, Sally Sent: Thursday, November 15, 2007 11:43 AM To: Robyn Vazquez; laurie.colbert@huntingtonhospital.com; histonet@lists.utsouthwestern.edu; mickie25@netzero.net Subject: RE: [Histonet] Colored OCT I have to ask the question. For what purpose do you use colored OCT? We here, at Children's in Seattle, have never heard of it. Thank you. Sally Norton, HT Children's Hospital and Regional Medical Center Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Thursday, November 15, 2007 6:29 AM To: laurie.colbert@huntingtonhospital.com; histonet@lists.utsouthwestern.edu; mickie25@netzero.net Subject: RE: [Histonet] Colored OCT Mickey, We like lots of color so we use 4-6 drops, depending on the color we want to achieve. BRIGHT COLORS!! And the variety of colors! It is amazing the little things that make us happy!!! Robyn OHSU _______________________________________________ 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 and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From josephnerk <@t> hotmail.com Thu Nov 15 12:32:40 2007 From: josephnerk <@t> hotmail.com (Joseph Nerk) Date: Thu Nov 15 12:32:50 2007 Subject: [Histonet] RE: Histonet Digest, Vol 48, Issue 21 In-Reply-To: References: Message-ID: Does anyone in Histoland provide any information on the Cryotome E or SME manufactured by Shandon. I am in the process of purchasing a new Cryostat. so need some feedback. > From: histonet-request@lists.utsouthwestern.edu> Subject: Histonet Digest, Vol 48, Issue 21> To: histonet@lists.utsouthwestern.edu> Date: Thu, 15 Nov 2007 10:00:16 -0800> > 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. WG: [Histonet] MyoD1 (Gudrun Lang)> 2. tissue processors (Jennifer Sipes)> 3. Hi all (Kimberly Secrest)> 4. RE: microtomy technical question (Monson, Frederick )> 5. Re: Giemsa Stain (Robert Richmond)> 6. pepsin (MKing)> 7. Re: pepsin (Joanne Mauger)> 8. RE: Colored OCT (Norton, Sally)> 9. Colored OCT (Webb, Dorothy L)> 10. Re: Giemsa Stain (Bill)> 11. Gill's III stainer question (karenadams@comcast.net)> > > ----------------------------------------------------------------------> > Message: 1> Date: Thu, 15 Nov 2007 16:12:16 +0100> From: "Gudrun Lang" > Subject: WG: [Histonet] MyoD1> To: > Message-ID: <001601c82799$e94a2850$6412a8c0@dielangs.at>> Content-Type: text/plain; charset="iso-8859-1"> > > 1:30, cc1 32 min, 32 min ink., 37?C > > Gudrun Lang> > Biomed. Analytikerin> Histolabor> Akh Linz> Krankenhausstr. 9> 4020 Linz> +43(0)732/7806-6754> -----Urspr?ngliche Nachricht-----> Von: histonet-bounces@lists.utsouthwestern.edu> [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von San Tin> Gesendet: Dienstag, 13. November 2007 02:35> An: histonet@lists.utsouthwestern.edu> Betreff: [Histonet] MyoD1> > Dear All,> > Can anyone give me the protocol of MyoD1 clone 5.8A on Ventana IHC> autostainer? I have problem with staining.> > > > Thanks> > San> > _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet> > > > > ------------------------------> > Message: 2> Date: Thu, 15 Nov 2007 07:19:13 -0800 (PST)> From: Jennifer Sipes > Subject: [Histonet] tissue processors> To: histonet@lists.utsouthwestern.edu> Message-ID: <98496.35382.qm@web62411.mail.re1.yahoo.com>> Content-Type: text/plain; charset=us-ascii> > I was wondering if anyone had a good protocol for mouse tissue on a Shandon Excelsior Tissue Processor.> > Thanks> > Jennifer K. Sipes, ALAT > Sr. Laboratory Technician > Johns Hopkins University > Ross 929 > 720 Rutland Avenue > Baltimore, MD 21205 > phone: 410-614-0131 > 410-955-9688 > fax: 410-955-9677 > cell: 443-631-6361 > e-mail: jsipes1@jhmi.edu> > > ____________________________________________________________________________________> Be a better pen pal. > Text or chat with friends inside Yahoo! Mail. See how. http://overview.mail.yahoo.com/> > ------------------------------> > Message: 3> Date: Thu, 15 Nov 2007 10:37:02 -0500> From: "Kimberly Secrest" > Subject: [Histonet] Hi all> To: > Message-ID: <473C2154.C068.0078.0@hsc.wvu.edu>> Content-Type: text/plain; charset=US-ASCII> > Does anybody have or know of anyone willing to donate histology equipment (i.e. microtomes, tissue processor, embedding center, routine stainer, etc) or supplies (i.e. slides, cassettes, reagents) for the start up of a histotechnology program?> > > Kimberly Secrest, HTL, QIHC> Instructor> Department of Pathology> School of Medicine> West Virginia University> > > > > > ------------------------------> > Message: 4> Date: Thu, 15 Nov 2007 10:57:31 -0500> From: "Monson, Frederick " > Subject: RE: [Histonet] microtomy technical question> To: > Message-ID:> <641CEFFC7E5B6C42AB59539653FD082304A90A58@wcu-ex-emp2.PASSHE.LCL>> Content-Type: text/plain; charset="us-ascii"> > I never respond without an answer from outside the bag, so to speak.> > 1. I once was quite ill (at the age of 40) and received the news that> my CBC required a bone marrow follow up. I asked if the differential> was at fault and heard a 'Yes'. I asked if the differential was> performed manually. The answer was 'No'. I demanded a manual> differential. The bone marrow was cancelled. Don't trust machines too> far is one of my life lessons.> > 2. I never had to use watered or iced paraffin blocks BEFORE I was> required by volume to process tissues in an auto processor. Prior to> using auto, I had very early on learned the consequences of> over-dehydrating, over-clearing and especially overcooking my specimens> and avoided doing so. Now with so many 110V AC lines measuring 128V AC> or <110V AC on occasion (the latter does no harm to the tissue but> lengthen the times in each bath), why is there a question about problems> with the outcomes?> > 3. For my SEM, I have a $5,600 Uninterruptible Power Supply (UPS) that> regulates the AC to within the specs of my instrument so that I am not> at the mercy of PECO - our local supplier - when I run long-duration> automated analyses.> > Most institutions are treated by the power companies as industrial> customers. The consequence is that power fluctuations are much more> frequent. Industry compensates by investing in regulation wherever it> is required. Institutions, on the other hand, are often ignorant of how> much or little power fluctuations affect performance in various> organizational modules - like laboratories in which analyses are> performed. One will not see the difference between 128 and 108 in the> performance of the coffee maker. Overnight, no one is watching and> monitoring.> > Hope this helps,> > Fred Monson> > Frederick C. Monson, PhD> Technical Director> Microanalysis and Imaging Research and Training Center (MIRTC)> Large Scientific Instrument Core> Geology, West Chester University> S. Church St. and W. Rosedale Ave.> West Chester, PA, 19320> 610-738-0437> fmonson@wcupa.edu> New Scheduler: http://lexspiac.wcupa.edu/cgi-bin/ureserve_gold.pl> Web Page: http://lexspiac.wcupa.edu> > > > > -----Original Message-----> From: histonet-bounces@lists.utsouthwestern.edu> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eva C> Andersson> Sent: Tuesday, November 13, 2007 4:32 PM> To: histonet@lists.utsouthwestern.edu> Subject: [Histonet] microtomy technical question> > Hello everyone,> I have some questions regarding microtomy. > We place our blocks face down on ice. The problem is that some tissues> take a very long time before they are ready to be cut(some more than 6> hours). For some tissues like kidney samples (mouse tissue) we have been> using glycerol on the ice. This does seem to cut down on the time> needed. My question is which other tissues can I use this technique on?> Do you have any other suggestions for how to get the tissue hydrated for> cutting?> Thank you for your help,> Eva Permaul> Georgetown University> > _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet> > > > ------------------------------> > Message: 5> Date: Thu, 15 Nov 2007 11:13:12 -0500> From: "Robert Richmond" > Subject: [Histonet] Re: Giemsa Stain> To: histonet@lists.utsouthwestern.edu> Message-ID:> > Content-Type: text/plain; charset=ISO-8859-1> > Bill (good ol' Bill - where?) says of staining Helicobacter:> > >>I like Alcian yellow better, but we have trouble getting it. Anyone> know of a good, reliable source in the US? I think IHC is overkill and> too expensive.<<> > The original chemical synthesis of Alcian yellow is not> environmentally safe, though the dye may still be made in countries> that are not concerned about such matters. Dick Dapson at Anatech told> us some time ago that he had achieved an environmentally acceptable> synthesis of Alcian yellow, but that the product did not have a> satisfactory shelf life. Anatech offers a substitute for Alcian> yellow, which I have not seen.> > (I have no connection with Anatech.)> > Bob Richmond> Samurai Pathologist> Knoxville TN> > > > ------------------------------> > Message: 6> Date: Thu, 15 Nov 2007 11:29:36 -0500> From: MKing > Subject: [Histonet] pepsin> To: histonet@lists.utsouthwestern.edu> Message-ID: <473C73F0.3010607@ufl.edu>> Content-Type: text/plain; charset=ISO-8859-1; format=flowed> > An antibody vendor recommended pepsin pretreatment prior to primary > antibody incubation of our free-floating 40 um formaldehyde-fixed rat > brain frozen sections, but provided a protocol that only listed pepsin > by weight. Since enzyme activity ranges from 3 to 4500 units/mg in > commercial sources, does anyone have a rough idea what kind of activity > is sufficient without destroying the tissue (which is what happened with > the first test). Anybody find this approach worthwhile with similar tissue?> Thanks,> Mike King> UF Pharmacology & Therapeutics> > > > ------------------------------> > Message: 7> Date: Thu, 15 Nov 2007 11:44:01 -0500> From: "Joanne Mauger" > Subject: Re: [Histonet] pepsin> To: ,> Message-ID: > Content-Type: text/plain; charset=US-ASCII> > Mike,> > We use pepsin at 2mgs/ml for digestion of formalin fixed, paraffin> embedded tissue. We use it at 37C for 10 to 30 minutes. We buy powdered> pepsin from Dako #S3002. > > Don't know what will happen to frozen tissue.> > Good luck,> > Jo Mauger> > > > ------------------------------> > Message: 8> Date: Thu, 15 Nov 2007 08:42:46 -0800> From: "Norton, Sally" > Subject: RE: [Histonet] Colored OCT> To: "Robyn Vazquez" ,> laurie.colbert@huntingtonhospital.com,> histonet@lists.utsouthwestern.edu, mickie25@netzero.net> Message-ID:> > Content-Type: text/plain; charset=us-ascii> > I have to ask the question. For what purpose do you use colored OCT?> We here, at Children's in Seattle, have never heard of it.> > Thank you.> Sally Norton, HT> Children's Hospital and Regional Medical Center> Seattle, WA> > > -----Original Message-----> From: histonet-bounces@lists.utsouthwestern.edu> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn> Vazquez> Sent: Thursday, November 15, 2007 6:29 AM> To: laurie.colbert@huntingtonhospital.com;> histonet@lists.utsouthwestern.edu; mickie25@netzero.net> Subject: RE: [Histonet] Colored OCT> > Mickey,> We like lots of color so we use 4-6 drops, depending on the color we> want to achieve. BRIGHT COLORS!! And the variety of colors! It is> amazing the little things that make us happy!!!> > Robyn> OHSU> _______________________________________________> 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 and privileged information protected by law. 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.> > > > > > ------------------------------> > Message: 9> Date: Thu, 15 Nov 2007 11:06:11 -0600> From: "Webb, Dorothy L" > Subject: [Histonet] Colored OCT> To: histonet@lists.utsouthwestern.edu> Message-ID:> <0E394B648E5284478A6CCB78E5AFDA270563528D@hpes1.HealthPartners.int>> Content-Type: text/plain; charset="us-ascii"> > TBS carries the colored OCT. The order number is H-TFM and is> available in red or blue.> > Dorothy Webb, HT (ASCP)> Histology Technical Supervisor > Regions Hospital, Pathology Department > 640 Jackson Street, Saint Paul, MN 55101-2595 > Phone: 651-254-2962> Fax: 651-254-2741 > Regions Hospital is part of the HealthPartners family of care> ________________________________________> This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited.> > If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us.> > > ------------------------------> > Message: 10> Date: Thu, 15 Nov 2007 11:27:14 -0600> From: Bill > Subject: [Histonet] Re: Giemsa Stain> To: histonet@lists.utsouthwestern.edu> Message-ID: > Content-Type: text/plain; charset="us-ascii"> > At 11:13 AM -0500 11/15/07, Robert Richmond wrote:> >Bill (good ol' Bill - where?) says of staining Helicobacter:> > I wondered why. Thanks Bob.> > > -- > _____________________________> Bill Blank> http://kernunnos.com (Celtic studies and numismatics)> OBOD's Message board: http://www.druidry.org/board/dhp/> > > > > > > ------------------------------> > Message: 11> Date: Thu, 15 Nov 2007 17:44:56 +0000> From: karenadams@comcast.net> Subject: [Histonet] Gill's III stainer question> To: histonet@lists.utsouthwestern.edu> Message-ID:> <111520071744.2924.473C859800022C3F00000B6C22070206539C030E0B0E020A9D0E05@comcast.net>> > Content-Type: text/plain> > > We are presently using Gill's III manually for H & E and on the automated stainer.....I am seeing mucin lightly staining on the automated stainer, but not the manual.....any thoughts??> --> Karen Adams > Supervisor> Pathology Laboratories West > 9303 Park West Blvd > Knoxville, TN 37923 > (865) 690-2111 FAX (865) 691-1623> > ------------------------------> > _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet> > End of Histonet Digest, Vol 48, Issue 21> **************************************** _________________________________________________________________ Discover the new Windows Vista http://search.msn.com/results.aspx?q=windows+vista&mkt=en-US&form=QBRE From jcline <@t> wchsys.org Thu Nov 15 12:43:23 2007 From: jcline <@t> wchsys.org (Joyce Cline) Date: Thu Nov 15 12:43:28 2007 Subject: [Histonet] Special Stainers Becky Johnson In-Reply-To: <00d901c82715$42e4da70$7b48f9d8@CHURCH> Message-ID: <001401c827b7$66935850$1d2a14ac@wchsys.org> Hi Becky, I use a Ventana stainer and I'll pass along a few hints that we have learned. For silver stains we use only Superfrost plus slides. I have tried some of the others but the staining quality was not adequate. We also leave our slides in the oven (72C) for 1/2 hour. We use the cleaning kit after every run. I don't know if you have tried any of these hints but if not, I hope they help. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca Johnson Sent: Wednesday, November 14, 2007 6:23 PM To: histonet Subject: [Histonet] Special Stainers I need to know what everyone is using for Special Stainers? I now have Ventana special stainer. Have had the same machine for several years with maintance(cleaning) done on it daily, monthly and PM. Just not real happy with the service and inconsistent staining with the silver stains. Thanks for all your info. Becky _______________________________________________ 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 Jackie.O'Connor <@t> abbott.com Thu Nov 15 12:52:07 2007 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Thu Nov 15 12:52:48 2007 Subject: [Histonet] Tape coverslipper In-Reply-To: <473C2154.C068.0078.0@hsc.wvu.edu> Message-ID: I love our automatic tape coverslipper. I'm a new user of an old machine, and I've noticed a problem. Perhaps someone can offer some insight. Occasionally, an artefact appears that resembles trapped air, microscopic bubbles, if you will, that appear gray-black on a higher plane than the tissue, so I know it's a superficial artefact. I just don't know what is causing it - but I'm sure it has something to do with the tape coverslipper - but what? I don't know enough about the machine to troubleshoot it - it seems a relatively simple instrument. Tape, a wheel and xylene. Where are the bubbles coming from, and how do I get rid of them? The pathologist thought it was some kind of dirt or dust debris accumulation - but we've decided it's air. Help. Jackie O' From JSiegel <@t> ucla.edu Thu Nov 15 12:52:48 2007 From: JSiegel <@t> ucla.edu (Jerry Siegel) Date: Thu Nov 15 12:53:09 2007 Subject: [Histonet] in situ and immunohistochemistry Message-ID: <6.2.1.2.2.20071115104638.0507a0e0@mail.ucla.edu> I would like combine in situ and immunohistochemistry on the same tissue, double labelling for mRNA and cell protein. I notice that published protocols use fixed tissue for the initial in situ step followed by immuno. We would ultimately like to perform this procedure on human tissue that has been in fix for years. We have been able to do immuno on this tissue using antigen retrieval techniques. Does anyone have any experience doing in situ on tissue that has been in fix for years? Does anyone recommend particular protocols for this double labelling procedure? J. Siegel From Jackie.O'Connor <@t> abbott.com Thu Nov 15 12:57:08 2007 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Thu Nov 15 12:57:48 2007 Subject: [Histonet] Tape coverslipper In-Reply-To: Message-ID: Oh yeah, I have pictures for anyone interested. Jackie O' Jackie M O'Connor Sent by: histonet-bounces@lists.utsouthwestern.edu 11/15/2007 12:52 PM To histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu cc Subject [Histonet] Tape coverslipper I love our automatic tape coverslipper. I'm a new user of an old machine, and I've noticed a problem. Perhaps someone can offer some insight. Occasionally, an artefact appears that resembles trapped air, microscopic bubbles, if you will, that appear gray-black on a higher plane than the tissue, so I know it's a superficial artefact. I just don't know what is causing it - but I'm sure it has something to do with the tape coverslipper - but what? I don't know enough about the machine to troubleshoot it - it seems a relatively simple instrument. Tape, a wheel and xylene. Where are the bubbles coming from, and how do I get rid of them? The pathologist thought it was some kind of dirt or dust debris accumulation - but we've decided it's air. Help. Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sally.norton <@t> seattlechildrens.org Thu Nov 15 12:59:20 2007 From: sally.norton <@t> seattlechildrens.org (Norton, Sally) Date: Thu Nov 15 12:59:35 2007 Subject: [Histonet] Colored OCT In-Reply-To: <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4F6@lmhsmail.lmhealth.org> References: <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4F6@lmhsmail.lmhealth.org> Message-ID: Thank you for the answers. We occasionally have more than one going at once, but not as often as you all do. Wow! Thanks, Sally -----Original Message----- From: Tom McNemar [mailto:TMcNemar@lmhealth.org] Sent: Thursday, November 15, 2007 10:33 AM To: Norton, Sally; Robyn Vazquez; laurie.colbert@huntingtonhospital.com; histonet@lists.utsouthwestern.edu; mickie25@netzero.net Subject: RE: [Histonet] Colored OCT We use the different colors when we have multiple frozens. Frozen 'A' is alwasy clear, 'B' is always blue, etc. I just helps to keep them straight. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Norton, Sally Sent: Thursday, November 15, 2007 11:43 AM To: Robyn Vazquez; laurie.colbert@huntingtonhospital.com; histonet@lists.utsouthwestern.edu; mickie25@netzero.net Subject: RE: [Histonet] Colored OCT I have to ask the question. For what purpose do you use colored OCT? We here, at Children's in Seattle, have never heard of it. Thank you. Sally Norton, HT Children's Hospital and Regional Medical Center Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Thursday, November 15, 2007 6:29 AM To: laurie.colbert@huntingtonhospital.com; histonet@lists.utsouthwestern.edu; mickie25@netzero.net Subject: RE: [Histonet] Colored OCT Mickey, We like lots of color so we use 4-6 drops, depending on the color we want to achieve. BRIGHT COLORS!! And the variety of colors! It is amazing the little things that make us happy!!! Robyn OHSU _______________________________________________ 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 and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information protected by law. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From rjbuesa <@t> yahoo.com Thu Nov 15 13:21:09 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 15 13:21:13 2007 Subject: [Histonet] Tape coverslipper In-Reply-To: Message-ID: <78801.1856.qm@web61222.mail.yahoo.com> Usually bubbles trapped underneath the tape are due to less than adequate amount of xylene. You have to regulate the size of the xylene drop to the amount needed to cover without bubbles. Don't overdue it because you may end with toomuch xylene and very "wet" slides. Ren? J. Jackie M O'Connor wrote: I love our automatic tape coverslipper. I'm a new user of an old machine, and I've noticed a problem. Perhaps someone can offer some insight. Occasionally, an artefact appears that resembles trapped air, microscopic bubbles, if you will, that appear gray-black on a higher plane than the tissue, so I know it's a superficial artefact. I just don't know what is causing it - but I'm sure it has something to do with the tape coverslipper - but what? I don't know enough about the machine to troubleshoot it - it seems a relatively simple instrument. Tape, a wheel and xylene. Where are the bubbles coming from, and how do I get rid of them? The pathologist thought it was some kind of dirt or dust debris accumulation - but we've decided it's air. Help. Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From mpence <@t> grhs.net Thu Nov 15 13:35:10 2007 From: mpence <@t> grhs.net (Mike Pence) Date: Thu Nov 15 13:35:20 2007 Subject: [Histonet] Tape coverslipper In-Reply-To: <78801.1856.qm@web61222.mail.yahoo.com> Message-ID: <661949901A768E4F9CC16D8AF8F2838CA1C7B2@IS-E2K3.grhs.net> To add to Ren?'s message: The xylene should drop on the slide like a string of pearls. One drop just not quit touching the other. Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Thursday, November 15, 2007 1:21 PM To: Jackie M O'Connor; histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Tape coverslipper Usually bubbles trapped underneath the tape are due to less than adequate amount of xylene. You have to regulate the size of the xylene drop to the amount needed to cover without bubbles. Don't overdue it because you may end with toomuch xylene and very "wet" slides. Ren? J. Jackie M O'Connor wrote: I love our automatic tape coverslipper. I'm a new user of an old machine, and I've noticed a problem. Perhaps someone can offer some insight. Occasionally, an artefact appears that resembles trapped air, microscopic bubbles, if you will, that appear gray-black on a higher plane than the tissue, so I know it's a superficial artefact. I just don't know what is causing it - but I'm sure it has something to do with the tape coverslipper - but what? I don't know enough about the machine to troubleshoot it - it seems a relatively simple instrument. Tape, a wheel and xylene. Where are the bubbles coming from, and how do I get rid of them? The pathologist thought it was some kind of dirt or dust debris accumulation - but we've decided it's air. Help. Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Thu Nov 15 13:50:49 2007 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Nov 15 13:51:16 2007 Subject: [Histonet] Colored OCT Message-ID: The reason we use it is, that it helps distinguish between different patients, while we are doing Mohs surgery. It is also appeasing and stimulating to the eye versus plain old white. Robyn OHSU From fudo <@t> ufl.edu Thu Nov 15 14:27:25 2007 From: fudo <@t> ufl.edu (FU,DONGTAO) Date: Thu Nov 15 14:27:39 2007 Subject: [Histonet] which species for stromal cell? Message-ID: <2021756729.220611195158445427.JavaMail.osg@osgjas03.cns.ufl.edu> Hi, You can use DAKOCytomation ARK kit to label your mouse antibody and do IHC staining on Xenograft tissue. Or you can use MOM kit from Vector to do the staining. Both are work fine in our lab. But if you want to detect stromal cells. You need use anti-mouse antibody. Because stromal cells come from mouse. Ann Dongtao Fu MD, Ph.D Lab Manager Molecular Pathology Core Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 On Thu Nov 15 13:14:24 EST 2007, Amy Lee wrote: > Hello, > I have an antibody only recognize human tissue. Now I have a > tumor tissue that is human tumor cell grow in mouse. I need to > stain stromal cell. Can I use this antibody? Or I may ask the > question this way: the human tumor cell grow in mouse, then is > the stromal cell develope from human tumor or from mouse tissue? > Thanks a lot! > Amy > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo > Mobile. Try it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From FARNANA <@t> nehealth.com Thu Nov 15 16:29:18 2007 From: FARNANA <@t> nehealth.com (Amy Farnan) Date: Thu Nov 15 16:29:32 2007 Subject: [Histonet] H&E stainer Message-ID: <473C81EE020000D900022B34@neh_domain_app_srv.nehealth.com> Hi folks - I am working at a small lab that is just getting started. We are looking to get an good but fairly inexpensive stainer for H&E slides. We are strained on space. At NSH I was looking at the Surgipath linear stainer. It is very basic and will fit well with our set up. We do not need anything fancy, just a basic stainer. Has anyone used this stainer and if so how well does it stain and hold up? If there are any other smaller stainers out there please let me know. Amy Farnan NEHealth Disclaimer: The information in this message is confidential. If you are not the intended recipient, do not disclose, copy, or distribute this message, and please immediately contact the sender. From jdmd77 <@t> hotmail.com Thu Nov 15 16:38:30 2007 From: jdmd77 <@t> hotmail.com (Julia Dahl) Date: Thu Nov 15 16:38:35 2007 Subject: [Histonet] Protocol for Rhodanine stain for copper Message-ID: Colleagues - Does anyone have a reliable protocol/procedure for the Rhodanine stain for bile that you would be willing to share? Thank you, Julia Dahl, M.D. _________________________________________________________________ Climb to the top of the charts!? Play Star Shuffle:? the word scramble challenge with star power. http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_oct From jacobc <@t> mmc.org Thu Nov 15 16:39:58 2007 From: jacobc <@t> mmc.org (Christine Jacobs) Date: Thu Nov 15 16:40:13 2007 Subject: [Histonet] MyoD1 Message-ID: <20071115T173958Z_C07000110000@mmc.org> I noticed a few other Histonetters with problems with their MyoD1. Here's my story. I use the Dako Cytomation concentrate-clone 5.8A. ON the Ventana Benchmark, my rpotocol is CC1-Mild, 60 minute incubation at 42 degrees (increased heat). I was using the antibody at a 1:100 dilution but when I started seeing poor/weak staining, I tried 1:50 and 1:20. My results were basically the same from one dilution to the other. There was varying degrees of background staining in both Rhabdo and skeletal muscle tissue and very rare cytoplasmic staining in the Rhabdo's. This is supposed to be a nuclear stain. I'm not sure if I have ever had good, consistent nuclear staining at any dilution. Any thoughts, comments, etc?........................ Chris Jacobs, IHC Specialist NorDx Labs Scarborough, ME CONFIDENTIALITY NOTICE: This email message, including any attachments, is for the use of the intended recipient(s) only and may contain information that is privileged, confidential, and prohibited from unauthorized disclosure under applicable law. If you are not the intended recipient of this message, any dissemination, distribution, or copying of this message is strictly prohibited. If you received this message in error, please notify the sender by reply email and destroy all copies of the original message and attachments. From mpence <@t> grhs.net Thu Nov 15 16:43:56 2007 From: mpence <@t> grhs.net (Mike Pence) Date: Thu Nov 15 16:44:02 2007 Subject: [Histonet] MyoD1 In-Reply-To: <20071115T173958Z_C07000110000@mmc.org> Message-ID: <661949901A768E4F9CC16D8AF8F2838CA1C7B7@IS-E2K3.grhs.net> What detection kit are you using? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Christine Jacobs Sent: Thursday, November 15, 2007 4:40 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] MyoD1 I noticed a few other Histonetters with problems with their MyoD1. Here's my story. I use the Dako Cytomation concentrate-clone 5.8A. ON the Ventana Benchmark, my rpotocol is CC1-Mild, 60 minute incubation at 42 degrees (increased heat). I was using the antibody at a 1:100 dilution but when I started seeing poor/weak staining, I tried 1:50 and 1:20. My results were basically the same from one dilution to the other. There was varying degrees of background staining in both Rhabdo and skeletal muscle tissue and very rare cytoplasmic staining in the Rhabdo's. This is supposed to be a nuclear stain. I'm not sure if I have ever had good, consistent nuclear staining at any dilution. Any thoughts, comments, etc?........................ Chris Jacobs, IHC Specialist NorDx Labs Scarborough, ME CONFIDENTIALITY NOTICE: This email message, including any attachments, is for the use of the intended recipient(s) only and may contain information that is privileged, confidential, and prohibited from unauthorized disclosure under applicable law. If you are not the intended recipient of this message, any dissemination, distribution, or copying of this message is strictly prohibited. If you received this message in error, please notify the sender by reply email and destroy all copies of the original message and attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ernestinemiddleton <@t> yahoo.ca Thu Nov 15 20:25:25 2007 From: ernestinemiddleton <@t> yahoo.ca (Ernestine Middleton) Date: Thu Nov 15 20:25:29 2007 Subject: [Histonet] (no subject) Message-ID: <494913.16935.qm@web51511.mail.re2.yahoo.com> Hi; Are any of lab using the cleaning cycle in the processor for cleaning their metal mold? Please comment . --------------------------------- Ask a question on any topic and get answers from real people. Go to Yahoo! Answers. From AnthonyH <@t> chw.edu.au Thu Nov 15 20:40:33 2007 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Thu Nov 15 20:41:50 2007 Subject: [Histonet] (no subject) Message-ID: We have now and then, but I often ask my staff "Why"? Are the metal molds any better to use? My opinion is that it is a waste of time, ie I have not noted any difference. 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: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ernestine Middleton Sent: Friday, 16 November 2007 1:25 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] (no subject) Hi; Are any of lab using the cleaning cycle in the processor for cleaning their metal mold? Please comment . --------------------------------- Ask a question on any topic and get answers from real people. Go to Yahoo! Answers. _______________________________________________ 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 note 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 jnocito <@t> satx.rr.com Fri Nov 16 06:07:59 2007 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Nov 16 06:08:08 2007 Subject: [Histonet] Colored OCT, Part II References: <130977.43220.qm@web63409.mail.re1.yahoo.com> Message-ID: <00d201c82849$553c7bd0$0302a8c0@yourxhtr8hvc4p> hey, do you have a MSDS on that food coloring? Is it being used according to manufactures recommendations? Has it been tested for phosphorus content? Have you performed an air sampling while that chemical is in use? It must be Friday. It must be Friday. Need more coffee, Need more coffee. There I go again, repeating myself. JTT ----- Original Message ----- From: "Matt Bancroft" To: ; "'Laurie Colbert'" ; Sent: Thursday, November 15, 2007 7:41 AM Subject: RE: [Histonet] Colored OCT >I use Neg 50 by Richard Allen > > Mickie Johnson wrote: Dear Laurie > > The OCT made by Sakura does not come in colors, but a drop of food > coloring > and mixing by rotating the bottle end over end will do the trick. If you > want darker color, use two drops. There are other brands of FS embedding > media out there which do come in colors, but I like OCT the best. > > Mickie > > Mickie Johnson, B.S., HTL(ASCP) > Mohs Histology Consulting Services, LLC > & Mohs Lab Staffing > 2507 S. Manito Blvd. > Spokane, WA 99203 > 509-954-7134 > Web: www.mohshistotemp.com & www.mohslabstaffing.com > Email: mickie25@netzero.net > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie > Colbert > Sent: Wednesday, November 14, 2007 2:54 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Colored OCT > > Where can I find OCT in different colors? I've seen it somewhere, but > now I can't find the vendor/manufacturer. > > > > Laurie Colbert > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try > it now. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lblazek <@t> digestivespecialists.com Fri Nov 16 07:02:34 2007 From: lblazek <@t> digestivespecialists.com (Blazek, Linda) Date: Fri Nov 16 06:59:02 2007 Subject: [Histonet] Colored OCT, Part II In-Reply-To: <00d201c82849$553c7bd0$0302a8c0@yourxhtr8hvc4p> References: <130977.43220.qm@web63409.mail.re1.yahoo.com> <00d201c82849$553c7bd0$0302a8c0@yourxhtr8hvc4p> Message-ID: <1F937FB30BDB7C4A9F39F83FEA8D379F3E4D43@bruexchange1.digestivespecialists.com> Especially if you use red. Linda Linda Blazek HT (ASCP) Manager/Supervisor GI Pathology of Dayton 7415 Brandt Pike Huber Heights, OH 45424 Phone: (937) 293-4424 ext 7118 Email: lblazek@digestivespecialists.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Friday, November 16, 2007 7:08 AM To: Matt Bancroft; mickie25@netzero.net; 'Laurie Colbert'; histonet@lists.utsouthwestern.edu Subject: [Histonet] Colored OCT, Part II hey, do you have a MSDS on that food coloring? Is it being used according to manufactures recommendations? Has it been tested for phosphorus content? Have you performed an air sampling while that chemical is in use? It must be Friday. It must be Friday. Need more coffee, Need more coffee. There I go again, repeating myself. JTT ----- Original Message ----- From: "Matt Bancroft" To: ; "'Laurie Colbert'" ; Sent: Thursday, November 15, 2007 7:41 AM Subject: RE: [Histonet] Colored OCT >I use Neg 50 by Richard Allen > > Mickie Johnson wrote: Dear Laurie > > The OCT made by Sakura does not come in colors, but a drop of food > coloring > and mixing by rotating the bottle end over end will do the trick. If you > want darker color, use two drops. There are other brands of FS embedding > media out there which do come in colors, but I like OCT the best. > > Mickie > > Mickie Johnson, B.S., HTL(ASCP) > Mohs Histology Consulting Services, LLC > & Mohs Lab Staffing > 2507 S. Manito Blvd. > Spokane, WA 99203 > 509-954-7134 > Web: www.mohshistotemp.com & www.mohslabstaffing.com > Email: mickie25@netzero.net > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie > Colbert > Sent: Wednesday, November 14, 2007 2:54 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Colored OCT > > Where can I find OCT in different colors? I've seen it somewhere, but > now I can't find the vendor/manufacturer. > > > > Laurie Colbert > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try > it now. > _______________________________________________ > Histonet mailing list > Histonet@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 rjbuesa <@t> yahoo.com Fri Nov 16 07:24:14 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 16 07:24:21 2007 Subject: [Histonet] (no subject) In-Reply-To: <494913.16935.qm@web51511.mail.re2.yahoo.com> Message-ID: <704240.49150.qm@web61225.mail.yahoo.com> Daily, as a standard procedure, the HT embedding waited until s/he finished, gathered all the molds, place them in the metal cassettes baskets and into the tissue processor and started the cleaning cycle. We always preferred to start embedding the next day with clean molds, without any paraffin particles attached. They look much better and is something obteinable without any additional reagent expense or time. Ren? J. Ernestine Middleton wrote: Hi; Are any of lab using the cleaning cycle in the processor for cleaning their metal mold? Please comment . --------------------------------- Ask a question on any topic and get answers from real people. Go to Yahoo! Answers. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From b-frederick <@t> northwestern.edu Fri Nov 16 07:46:21 2007 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Fri Nov 16 07:46:33 2007 Subject: [Histonet] (no subject) In-Reply-To: <494913.16935.qm@web51511.mail.re2.yahoo.com> Message-ID: <001901c82857$15394640$d00f7ca5@lurie.northwestern.edu> Every day. We change the cleaning solutions weekly also. Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ernestine Middleton Sent: Thursday, November 15, 2007 8:25 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] (no subject) Hi; Are any of lab using the cleaning cycle in the processor for cleaning their metal mold? Please comment . --------------------------------- Ask a question on any topic and get answers from real people. Go to Yahoo! Answers. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mpence <@t> grhs.net Fri Nov 16 08:09:58 2007 From: mpence <@t> grhs.net (Mike Pence) Date: Fri Nov 16 08:10:10 2007 Subject: [Histonet] (no subject) In-Reply-To: <494913.16935.qm@web51511.mail.re2.yahoo.com> Message-ID: <661949901A768E4F9CC16D8AF8F2838CA1C7B9@IS-E2K3.grhs.net> We clean our molds and metal lids this way. Much better to start each day with clean lids and molds. Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ernestine Middleton Sent: Thursday, November 15, 2007 8:25 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] (no subject) Hi; Are any of lab using the cleaning cycle in the processor for cleaning their metal mold? Please comment . --------------------------------- Ask a question on any topic and get answers from real people. Go to Yahoo! Answers. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Dolores_Fischer <@t> baxter.com Fri Nov 16 08:43:32 2007 From: Dolores_Fischer <@t> baxter.com (Dolores_Fischer@baxter.com) Date: Fri Nov 16 08:51:29 2007 Subject: [Histonet] immunohistochemistry on drugs Message-ID: Good morning! Has anyone out there is histoland ever tried detecting a specific drug in tissue using an immunohistochemical procedure? Not a biological marker that the drug may attach to, but the drug directly? Thanks for any info! Dolores Fischer The information transmitted is intended only for the person(s)or entity to which it is addressed and may contain confidential and/or legally privileged material. Delivery of this message to any person other than the intended recipient(s) is not intended in any way to waive privilege or confidentiality. Any review, retransmission, dissemination or other use of , or taking of any action in reliance upon, this information by entities other than the intended recipient is prohibited. If you receive this in error, please contact the sender and delete the material from any computer. For Translation: http://www.baxter.com/email_disclaimer From carmen_loiselle <@t> hotmail.com Fri Nov 16 08:54:14 2007 From: carmen_loiselle <@t> hotmail.com (carmen loiselle) Date: Fri Nov 16 08:54:20 2007 Subject: [Histonet] TFE-3 Message-ID: Can anybody share their protocol for this antibody ? I'm currently using the Benchmark and XT from Ventana. I tried different dilutions as well as different incubation time, different antigen retrieval solution (CC1 and CC2) and time.. Any input would be greatly appreciated . Thanks _________________________________________________________________ Envoie un sourire, fais rire, amuse-toi! Employez-le maintenant! http://www.emoticonesgratuites.ca/?icid=EMFRCA120 From gu.lang <@t> gmx.at Fri Nov 16 09:01:07 2007 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Nov 16 09:01:15 2007 Subject: AW: [Histonet] (no subject) In-Reply-To: <494913.16935.qm@web51511.mail.re2.yahoo.com> Message-ID: <000301c82861$850a1b70$6412a8c0@dielangs.at> We clean our molds with an ultrasonic bath. 15 min with a drop of dishwasher. Every day. Gudrun Lang Biomed. Analytikerin Histolabor Akh Linz Krankenhausstr. 9 4020 Linz +43(0)732/7806-6754 -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Ernestine Middleton Gesendet: Freitag, 16. November 2007 03:25 An: 'histonet@pathology.swmed.edu' Betreff: [Histonet] (no subject) Hi; Are any of lab using the cleaning cycle in the processor for cleaning their metal mold? Please comment . --------------------------------- Ask a question on any topic and get answers from real people. Go to Yahoo! Answers. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eca9 <@t> georgetown.edu Fri Nov 16 09:29:22 2007 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Nov 16 09:29:32 2007 Subject: [Histonet] microtomy technical question: Protocol used for processing Message-ID: Hello again, I recently posted a question regarding hydrating samples before cutting them. I am having the problem that my samples are very dry and have had to use glycerol on several of my different tissues. I have asked the technician who does the processing about the protocol that is being used as many of you have said that the samples are probably being overprocessed. They usually use one program but sometimes a slightly shorter one. Problem has been that in the past some tissues turned up underprocessed. Most of the tissues we process are mouse and rat. This is what they use: (in a Leica ASP300 processor) 70% Ethanol 55min 80% Ethanol 55min 95% Ethanol 55min 95% Ethanol 1hour 100% Ethanol 55min 100% Ethanol 1hour Clear rite 55min Clear rite 55min Clear rite 1hour Paraffin Wax 1hour Paraffin Wax 1hour Paraffin Wax 1hour Do you have any helpful suggestions? Very tired of having to spend all day waiting on my samples to hydrate. Thank in advance, Eva Permaul Georgetown University From rjbuesa <@t> yahoo.com Fri Nov 16 09:57:14 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 16 09:57:17 2007 Subject: [Histonet] microtomy technical question: Protocol used for processing In-Reply-To: Message-ID: <895094.48132.qm@web61223.mail.yahoo.com> Cut your dehydrating and clearing times in half. Eliminate one 95% EthOL and add one station with 100 EthOL-clearing 1:2 instead (after the last 100% EthOL). Ren? J. Eva C Andersson wrote: Hello again, I recently posted a question regarding hydrating samples before cutting them. I am having the problem that my samples are very dry and have had to use glycerol on several of my different tissues. I have asked the technician who does the processing about the protocol that is being used as many of you have said that the samples are probably being overprocessed. They usually use one program but sometimes a slightly shorter one. Problem has been that in the past some tissues turned up underprocessed. Most of the tissues we process are mouse and rat. This is what they use: (in a Leica ASP300 processor) 70% Ethanol 55min 80% Ethanol 55min 95% Ethanol 55min 95% Ethanol 1hour 100% Ethanol 55min 100% Ethanol 1hour Clear rite 55min Clear rite 55min Clear rite 1hour Paraffin Wax 1hour Paraffin Wax 1hour Paraffin Wax 1hour Do you have any helpful suggestions? Very tired of having to spend all day waiting on my samples to hydrate. Thank in advance, Eva Permaul Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Never miss a thing. Make Yahoo your homepage. From TJJ <@t> Stowers-Institute.org Fri Nov 16 09:57:16 2007 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Nov 16 09:57:40 2007 Subject: [Histonet] Matrigel as culture medium Message-ID: One of our researchers is using matrigel as a culture medium on chambered slides. We tried paraffin embedding them, and found the Matrigel wasn't really solid enough to hold the cellular structure in place. We found it particularly difficult to keep some sort of consistency scraping it off (it had the consistency of warm jelly). I'd love to hear some ideas on how we can accomplish sample processing without centrifugation. Ideally it would be good to encase the cells in a matrix that would solidfy enough to embed as a cell block, keeping the cellular structure intact. Thanks for your help! Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 From mpence <@t> grhs.net Fri Nov 16 10:09:59 2007 From: mpence <@t> grhs.net (Mike Pence) Date: Fri Nov 16 10:10:08 2007 Subject: [Histonet] microtomy technical question: Protocol used forprocessing In-Reply-To: Message-ID: <661949901A768E4F9CC16D8AF8F2838CA1C7BB@IS-E2K3.grhs.net> Give the fact that your tissue most likely contain very little fibroadipose tissue and that your gross tissue sections are most likely half of what a human tissue is. I would say that your are over processing your tissue by at least twice. Cut your ETOH times in half to start and get rid of one 95% - cut your clear rite in half and add another station of clear rite. This is where I wold start. Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eva C Andersson Sent: Friday, November 16, 2007 9:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] microtomy technical question: Protocol used forprocessing Hello again, I recently posted a question regarding hydrating samples before cutting them. I am having the problem that my samples are very dry and have had to use glycerol on several of my different tissues. I have asked the technician who does the processing about the protocol that is being used as many of you have said that the samples are probably being overprocessed. They usually use one program but sometimes a slightly shorter one. Problem has been that in the past some tissues turned up underprocessed. Most of the tissues we process are mouse and rat. This is what they use: (in a Leica ASP300 processor) 70% Ethanol 55min 80% Ethanol 55min 95% Ethanol 55min 95% Ethanol 1hour 100% Ethanol 55min 100% Ethanol 1hour Clear rite 55min Clear rite 55min Clear rite 1hour Paraffin Wax 1hour Paraffin Wax 1hour Paraffin Wax 1hour Do you have any helpful suggestions? Very tired of having to spend all day waiting on my samples to hydrate. Thank in advance, Eva Permaul Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mari.ann.mailhiot <@t> leica-microsystems.com Fri Nov 16 10:27:02 2007 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Nov 16 10:27:19 2007 Subject: [Histonet] microtomy technical question: Protocol used for processing In-Reply-To: Message-ID: Hi Eva You can cut your times down to around 30 minutes for each station. You are taking the bound water out of the specimens. You need that water to keep the specimens from being too dry. That being said animal tissue is dry to begin with. There is an excellent Animal Tissue Processing Manual that was published for NSH and edited by Gayle Callis and Diane Sterchi. This is an excellent resource. I have used it many times. The manual can be purchased through NSH. If you need further assistance please don't hesitate to give me a call. Best Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com Eva C Andersson To Sent by: histonet@lists.utsouthwestern.edu histonet-bounces@ cc lists.utsouthwest ern.edu Subject [Histonet] microtomy technical question: Protocol used for 11/16/2007 09:29 processing AM Hello again, I recently posted a question regarding hydrating samples before cutting them. I am having the problem that my samples are very dry and have had to use glycerol on several of my different tissues. I have asked the technician who does the processing about the protocol that is being used as many of you have said that the samples are probably being overprocessed. They usually use one program but sometimes a slightly shorter one. Problem has been that in the past some tissues turned up underprocessed. Most of the tissues we process are mouse and rat. This is what they use: (in a Leica ASP300 processor) 70% Ethanol 55min 80% Ethanol 55min 95% Ethanol 55min 95% Ethanol 1hour 100% Ethanol 55min 100% Ethanol 1hour Clear rite 55min Clear rite 55min Clear rite 1hour Paraffin Wax 1hour Paraffin Wax 1hour Paraffin Wax 1hour Do you have any helpful suggestions? Very tired of having to spend all day waiting on my samples to hydrate. Thank in advance, Eva Permaul Georgetown University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From PMonfils <@t> Lifespan.org Fri Nov 16 11:33:36 2007 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Nov 16 11:33:42 2007 Subject: [Histonet] microtomy technical question: Protocol used forprocessing In-Reply-To: Message-ID: <4EBFF65383B74D49995298C4976D1D5E273CF9@LSRIEXCH1.lsmaster.lifespan.org> The times given would be right for some types of samples, excessive for other types, and insufficient for still other types of samples. The principle factor is sample size, though tissue type also plays a role. If you are running small biopsies, only a few mm in size, 1/3 the listed time in each station should be adequate. For average tissue samples up to a cm or so in smallest dimension, the listed times are probably about right. For larger samples, especially those that are fatty or unusually dense, longer processing times would be recommended, though that doesn't seem to be your problem. There is no single processing schedule that is right for all tissue samples. Unfortunately, most clinical labs don't have much choice in this matter. Everything has to be run overnight and embedded in the morning. Where I work, in a core research facility, we can customize processing for the specific specimens we are working with. I have processing schedules programmed into the processor with total run times anywhere from 3 hours to 64 hours. And occasionally I run shorter or longer programs than these. From Heather.D.Renko <@t> osfhealthcare.org Fri Nov 16 11:57:43 2007 From: Heather.D.Renko <@t> osfhealthcare.org (Renko, Heather D.) Date: Fri Nov 16 11:58:00 2007 Subject: [Histonet] RE: cleaning cycle Message-ID: <40026EDDE64CDA47AB382C52619ACD3C0A09DC04@pmc-rfd-mx01.intranet.osfnet.org> We do a cleaning cycle daily on our metal base molds. Heather D. Renko, Histology Coordinator OSF Saint Anthony Medical Center 5666 East State Street Rockford, Illinois 61108 Direct: 815-395-5410 Heather.D.Renko@osfhealthcare.org ============================================================================== The information in this message is confidential and may be legally privileged. Access to this message by anyone other than the addressee is not authorized. If you are not the intended recipient, or an agent of the intended recipient, any disclosure, copying, or distribution of the message or any action or omission taken by you in reliance on it, is prohibited and may be unlawful. If you have received this message in error, please contact the sender immediately and permanently delete the original e-mail, attachment(s), and any copies. ============================================================================== From CIngles <@t> uwhealth.org Fri Nov 16 12:01:34 2007 From: CIngles <@t> uwhealth.org (Ingles Claire) Date: Fri Nov 16 12:05:25 2007 Subject: [Histonet] non pathologist grossing vs processing continued References: <44780C571F28624DBB446DE55C4D733A1FE56A@EXCHANGEBE1.carle.com> Message-ID: <08A0A863637F1349BBFD83A96B27A50A1200A1@uwhis-xchng3.uwhis.hosp.wisc.edu> Sorry this is a bit behind. I have been too busy to check my mail. Do you think this includes someone who has an AA in Electron Microscopy? I'm inclined to think so, but contrary to popular belief, I have been wrong before. "On or before April 24 1995 (I) be a high school graduate or equivalent; and (b) have documentation of training appropriate for the test performed before analyzing patient specimens"...After that date it requires an associate degree in a biological or chemical science or medical laboratory technology -or- qualify as a medical technologist with a bachelor's degree from an accredited institution -or- earned a bachelor's degree in a chemical, physical, biologic or clinical laboratory science. Thanks for the Advice, Claire From Dorothy.L.Webb <@t> HealthPartners.Com Fri Nov 16 12:34:08 2007 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Fri Nov 16 12:34:15 2007 Subject: [Histonet] Cleaning molds Message-ID: <0E394B648E5284478A6CCB78E5AFDA2705635298@hpes1.HealthPartners.int> At the previous facility I worked at, we daily cleaned our mold in the cleaning cycle until we had a problem with the processor and was told by the field service engineer and then the company that doing this was hard on the machine with all of the excess paraffin in the tubing. I do not know if that has any merit, but, we quit doing that and never had that type of problem again. Just a thought!! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From funderwood <@t> mcohio.org Fri Nov 16 12:43:18 2007 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Nov 16 12:43:45 2007 Subject: [Histonet] Cleaning molds In-Reply-To: <0E394B648E5284478A6CCB78E5AFDA2705635298@hpes1.HealthPartners.int> References: <0E394B648E5284478A6CCB78E5AFDA2705635298@hpes1.HealthPartners.int> Message-ID: <473D9E760200003400002C2F@mcohio.org> Hi Dorothy, Depending on the brand, some processors will heat the preheat the retort. Then pump it back into the last paraffin. The VIP does this, which is what I have. I clean my molds this way, once a week maximum, and have not had problems. I had an old MVP prior to the VIP and it did not pump back any molten paraffin. Fred >>> "Webb, Dorothy L" 11/16/2007 1:34 PM >>> At the previous facility I worked at, we daily cleaned our mold in the cleaning cycle until we had a problem with the processor and was told by the field service engineer and then the company that doing this was hard on the machine with all of the excess paraffin in the tubing. I do not know if that has any merit, but, we quit doing that and never had that type of problem again. Just a thought!! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Shirley.Chu <@t> moldev.com Fri Nov 16 13:08:32 2007 From: Shirley.Chu <@t> moldev.com (Chu, Shirley) Date: Fri Nov 16 13:08:42 2007 Subject: [Histonet] LCM Web Seminar Message-ID: The next in the series of LCM Web Seminars will be presented on Wednesday, Nov 28th at 1:30pm EDT/10:30am PDT. The speaker will be Virgina Espina, MS, MT(ASCP) of George Mason University, Center for Applied Proteomics and Molecular Medicine. The title of her talk is: "LCM for Clinical Proteomics Applications". For further information and to register for this free webinar, please see the following website: http://www.moleculardevices.com/pages/webinar_arcturus_11.2007.html Please also note that all previous LCM webinar presentations are available through this website. Shirley Chu Applications Scientist, Arcturus LCM Products Molecular Devices (now a part of MDS Analytical Technologies) 510-675-6260 | www.moleculardevices.com From Dorothy.L.Webb <@t> HealthPartners.Com Fri Nov 16 13:09:39 2007 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Fri Nov 16 13:09:46 2007 Subject: [Histonet] Minimum time processing Message-ID: <0E394B648E5284478A6CCB78E5AFDA2705635299@hpes1.HealthPartners.int> For those histology labs doing prostate needle biopsies.....I need some advice!! We process our pnbx on a separate program on a VIP 5 and these biospies are taken with a 20 gauge needle, very thin. What would be the minimum times you would recommend processing in the reagents? I am currently doing 10 minutes per station and I am seeing some very dry cores (they are usually the tiniest ones) that, of course, are staining very dark. I have a pathologist that feels we can do better, so needing my super histotechs to help me out once again!!! Thank you So much for help you always send via the histonet!! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From froyer <@t> bitstream.net Fri Nov 16 13:26:15 2007 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Nov 16 13:27:03 2007 Subject: [Histonet] Cleaning molds In-Reply-To: <0E394B648E5284478A6CCB78E5AFDA2705635298@hpes1.HealthPartners.int> References: <0E394B648E5284478A6CCB78E5AFDA2705635298@hpes1.HealthPartners.int> Message-ID: <007a01c82886$8e8049c0$7701a80a@Ford> >From the service side, I have experienced what Dorothy has described... especially on older model tissue processors. A good percentage of service calls are for plugged reagent, overflow, and/or air lines that have simply been overpowered by excessive amounts of paraffin during the cleaning cycle. This has been true even with processors that melt the residual paraffin first and pumps it back into the last paraffin station (i.e. VIP's). However in some of these cases, it should be noted that the amount of paraffin on molds, forceps, and other metal items that were placed in the retort for cleaning was very excessive. I am a believer that you can still use your processor as a mold cleaner if you limit the amount of residual paraffin on the molds (forceps, etc.) by scraping off as much as possible prior to placing them in the retort. You should also orient items so that melted paraffin can drip down to the bottom of the retort so it can actually be pumped out. It does no good to put molds and boats in the retort face up so that all the melted paraffin is simply held in the mold itself and not allowed to drain to the bottom of the retort to be pumped out. If used as a "dish washer", you should also change the "Cleaning Xylene" reagent more frequently. ~ Ford Ford M. Royer, MT(ASCP) Minnesota Medical, Inc. 7177 Madison Ave. W. Golden Valley, MN 55427-3601 CELL:? 612-839-1046 Phone:? 763-542-8725 Fax:? 763-546-4830 eMail:? clinicallab@minnesotamedical.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Webb, Dorothy L Sent: Friday, November 16, 2007 12:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cleaning molds At the previous facility I worked at, we daily cleaned our mold in the cleaning cycle until we had a problem with the processor and was told by the field service engineer and then the company that doing this was hard on the machine with all of the excess paraffin in the tubing. I do not know if that has any merit, but, we quit doing that and never had that type of problem again. Just a thought!! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Fri Nov 16 14:41:48 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 16 14:41:53 2007 Subject: [Histonet] Cleaning molds In-Reply-To: <0E394B648E5284478A6CCB78E5AFDA2705635298@hpes1.HealthPartners.int> Message-ID: <296355.57159.qm@web61225.mail.yahoo.com> I am sure you were "taken for a ride" with that opinion. There is no way the small amounts of paraffin left in the molds are going to significantly impact in the tubing, specially when the retort is hot and will melt it all. Even sometimes there is a remnant in the retorn (from the processing) that is usually more than what you can add with the cassettes. Who told you that was trying to blame you for something s/he could not know how to repair. But, in any event, you can do nothing about that now, just bear in mind for the next time you are told the same nonsense. Ren? J. "Webb, Dorothy L" wrote: At the previous facility I worked at, we daily cleaned our mold in the cleaning cycle until we had a problem with the processor and was told by the field service engineer and then the company that doing this was hard on the machine with all of the excess paraffin in the tubing. I do not know if that has any merit, but, we quit doing that and never had that type of problem again. Just a thought!! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From kellerc2 <@t> uthscsa.edu Fri Nov 16 14:42:54 2007 From: kellerc2 <@t> uthscsa.edu (Keller, Charles) Date: Fri Nov 16 14:42:58 2007 Subject: [Histonet] Matrigel as culture medium In-Reply-To: References: Message-ID: Dear Teri, a possible solution is to use Extracel (from Glycosan Inc in Utah) instead of Matrigel. I've fixed Extracel in 10% formalin and sectioned in in paraffin. It has the advantage of being synthetic (Matrigel has viral contamination from time to time). Sincerely, Charles -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Friday, November 16, 2007 9:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Matrigel as culture medium One of our researchers is using matrigel as a culture medium on chambered slides. We tried paraffin embedding them, and found the Matrigel wasn't really solid enough to hold the cellular structure in place. We found it particularly difficult to keep some sort of consistency scraping it off (it had the consistency of warm jelly). I'd love to hear some ideas on how we can accomplish sample processing without centrifugation. Ideally it would be good to encase the cells in a matrix that would solidfy enough to embed as a cell block, keeping the cellular structure intact. Thanks for your help! Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcline <@t> wchsys.org Fri Nov 16 16:08:21 2007 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Nov 16 16:08:30 2007 Subject: [Histonet] Special Stainers Becky Johnson In-Reply-To: <00db01c827f5$0d492f70$7b48f9d8@CHURCH> Message-ID: <000801c8289d$34992f50$1d2a14ac@wchsys.org> Becky, Our first stainer we had for 10 years, we received a new one this year. Ventana told us the pH should be 5.5 to 6.0 (-or+ .1). We check the pH of our special stain wash when we mix it, not at each run. -----Original Message----- From: Rebecca Johnson [mailto:raj@bluemarble.net] Sent: Thursday, November 15, 2007 9:05 PM To: Joyce Cline Subject: Re: [Histonet] Special Stainers Becky Johnson Joyce, We have always done everything Ventana told us to do from the very beginning. We use the plus green slides that Ventana advised. Daily before any run is started the Ph is taken on the special stain wash., the level is checked and the liquid coverslip, special stain wash filled then the machine is primed to clear all lines from any air or bubbles. The clean cycle is done when special stains are finished for the day. What has Ventana told you the Ph should be on the special stain wash? We only have one Nexus stainer and it is several years old, but the maintance has always been done regular. We have had the Benchmark XT for about a year. So far no problems with it. Thanks for all your advice. Becky ----- Original Message ----- From: "Joyce Cline" To: Sent: Thursday, November 15, 2007 1:43 PM Subject: RE: [Histonet] Special Stainers Becky Johnson > Hi Becky, > I use a Ventana stainer and I'll pass along a few hints that we have > learned. For silver stains we use only Superfrost plus slides. I have > tried some of the others but the staining quality was not adequate. We > also leave our slides in the oven (72C) for 1/2 hour. We use the > cleaning kit after every run. > I don't know if you have tried any of these hints but if not, I hope > they help. > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca > Johnson > Sent: Wednesday, November 14, 2007 6:23 PM > To: histonet > Subject: [Histonet] Special Stainers > > I need to know what everyone is using for Special Stainers? I now have > Ventana special stainer. Have had the same machine for several years > with maintance(cleaning) done on it daily, monthly and PM. Just not > real happy with the service and inconsistent staining with the silver > stains. Thanks for all your info. > Becky > _______________________________________________ > 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. > > > _______________________________________________ > 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 cmiller <@t> physlab.com Sat Nov 17 07:33:36 2007 From: cmiller <@t> physlab.com (Cheri Miller) Date: Sat Nov 17 07:34:04 2007 Subject: [Histonet] Cleaning molds In-Reply-To: <0E394B648E5284478A6CCB78E5AFDA2705635298@hpes1.HealthPartners.int> References: <0E394B648E5284478A6CCB78E5AFDA2705635298@hpes1.HealthPartners.int> Message-ID: <005101c8291e$74dd3170$3402a8c0@plab.local> What we do is, warm the dirty rack, lids and molds in the oven to rid of excess paraffin and then put them thru the clean cycle. Works well for us. It's not so hard on the processor, Cheri Cheri Miller HT ASCP Histology Supervisor Physicians Laboratory Services, Inc. Omaha, NE 68117 402 738 5052 PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Webb, Dorothy L Sent: Friday, November 16, 2007 12:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cleaning molds At the previous facility I worked at, we daily cleaned our mold in the cleaning cycle until we had a problem with the processor and was told by the field service engineer and then the company that doing this was hard on the machine with all of the excess paraffin in the tubing. I do not know if that has any merit, but, we quit doing that and never had that type of problem again. Just a thought!! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. From aakrasht <@t> yahoo.com Sat Nov 17 12:45:38 2007 From: aakrasht <@t> yahoo.com (Ali A. Krasht) Date: Sat Nov 17 12:45:44 2007 Subject: [Histonet] Need a sample business plan in Histology (HELP) Message-ID: <61356.25132.qm@web50902.mail.re2.yahoo.com> Hi There Anyone would like to help, I need a sample business plan for opening a Histology lab (human tissue in Dallas Texas) so I can have an idea in how to plan or write it specially the projection part is given me trouble; I got all the outline from the Small Business Bureau on how to write it but I need a little help by sending me a sample plan in Histology, I could not find any online. Thanks for your time on this matter. Ali ____________________________________________________________________________________ Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. http://overview.mail.yahoo.com/ From sheila_adey <@t> hotmail.com Sat Nov 17 14:41:02 2007 From: sheila_adey <@t> hotmail.com (sheila adey) Date: Sat Nov 17 14:41:08 2007 Subject: [Histonet] (no subject) Message-ID: Hi Netters, We are trying to minimize possible embedding contaminations. What are other people doing to prevent contamination due to forceps etc. Thanks in advanceSheila Adey HT MLTPort Huron HospitalMichigan _________________________________________________________________ Are you ready for Windows Live Messenger Beta 8.5 ? Get the latest for free today! http://entertainment.sympatico.msn.ca/WindowsLiveMessenger From sheila_adey <@t> hotmail.com Sat Nov 17 14:46:43 2007 From: sheila_adey <@t> hotmail.com (sheila adey) Date: Sat Nov 17 14:46:48 2007 Subject: [Histonet] RE: [Embedding contamination In-Reply-To: References: Message-ID: Sheila Adey HT MLTPort Huron HospitalMichigan> From: sheila_adey@hotmail.com> To: histonet@lists.utsouthwestern.edu> Date: Sat, 17 Nov 2007 15:41:02 -0500> Subject: [Histonet] (no subject)> > > Hi Netters,> > We are trying to minimize possible embedding contaminations. What are other people doing to prevent contamination due to forceps etc. > > Thanks in advanceSheila Adey HT MLTPort Huron HospitalMichigan> _________________________________________________________________> Are you ready for Windows Live Messenger Beta 8.5 ? Get the latest for free today!> http://entertainment.sympatico.msn.ca/WindowsLiveMessenger_______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Are you ready for Windows Live Messenger Beta 8.5 ? Get the latest for free today! http://entertainment.sympatico.msn.ca/WindowsLiveMessenger From rjbuesa <@t> yahoo.com Sat Nov 17 15:02:15 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Nov 17 15:02:20 2007 Subject: [Histonet] (no subject) In-Reply-To: Message-ID: <398204.57053.qm@web61215.mail.yahoo.com> Have several forceps and clean them frequently and, most important, thoroughly clean the wells in the embedding center each day; first with a dropper sucking out any debry, and finally with a Q-tip until completely clean. Ren? J. sheila adey wrote: Hi Netters, We are trying to minimize possible embedding contaminations. What are other people doing to prevent contamination due to forceps etc. Thanks in advanceSheila Adey HT MLTPort Huron HospitalMichigan _________________________________________________________________ Are you ready for Windows Live Messenger Beta 8.5 ? Get the latest for free today! http://entertainment.sympatico.msn.ca/WindowsLiveMessenger_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From swelam48 <@t> gmail.com Sat Nov 17 23:41:42 2007 From: swelam48 <@t> gmail.com (wael swelam) Date: Sat Nov 17 23:41:46 2007 Subject: [Histonet] Quick Decalcification Message-ID: <96566f7e0711172141q1b16306dy6ca69aebfc734351@mail.gmail.com> Dear Histonet members; Good morning to all of you. Do you have a formula that can be used to achieve quick decalcification for bone & Dental hard tissues without affecting the quality of the related soft tissues. Your kind help is highly appreciated Yours, Wael Mohamed Swelam BDS (Egypt), Oral Pathology MSc (Egypt), Oral Pathology PhD (Japan), Ass. Prof. Oral Biomedical Science dept. Division of Oral Pathology, Faculty of Dentistry, King Faisal University, Dammam, Saudi Arabia, P.O: 1982/3441, http://myprofile.cos.com/wmswelam From spotts <@t> aperio.com Sun Nov 18 03:12:01 2007 From: spotts <@t> aperio.com (Steven Potts) Date: Sun Nov 18 03:12:12 2007 Subject: [Histonet] RE: immunohistochemistry on drugs Message-ID: <52F0A1B1F834C54DB1C0CF32182D6E9B01B46F39@sagan.aperio.int> I?m also really curious if others have seen any protocols, using IHC for ADME type distribution tests. This area has typically the domain of radiolabeling protocols, IHC would have some advantages if an assay could be developed. What kind of drug (small molecule, biological, enzyme, antisense) would be the first question. I?ve found the following from a brief literature search: If the drug happens to be an enzyme, it can of course be done: For example, see Murray, GJ et al, Molecular Genetics and Metabolism Volume 90, Issue 3, March 2007, Pages 307-312 ?Cellular and tissue distribution of intravenously administered agalsidase alfa?. If your drug is an antisense compound, this is done regularly: For example, see Butler, M. Neuroscience 131(3): 2005. 705-715. Spinal distribution and metabolism of 2?-O-(2-methoxyethyl)-modified oligonucleotides after intrathecal administration in rats. Seems like you ought to be able to put a monoclonal antibody with DAB staining process specific for the drug, and see the distribution this way. Anyone know of an example of a small molecule drug compound that someone did this with IHC? Would it be as sensitive as direct radiolabelling? I?d like to know the answer, because with whole slide scanning and computer measurements of IHC on a cell by cell basis, we could do very consistent quantitative of the drug expression level in tissue, provided there was a colorimetric approach to correlated with drug concentration in the tissue of interest. Steve Potts, Ph.D. Scientific Director, Biopharma Aperio Technologies phone: (760) 539-1118 email: spotts@aperio.com web: www.aperio.com From kemlo <@t> f2s.com Sun Nov 18 04:21:46 2007 From: kemlo <@t> f2s.com (kemlo) Date: Sun Nov 18 04:22:10 2007 Subject: [Histonet] (no subject) In-Reply-To: References: Message-ID: In the good old days of Bunsen burners we used to fry the end of the forceps; nothing survived that and if it did it was readily identifiable. Alas the Bunsen burner has been consigned to the politically incorrect as the 'Scientists' of today would incinerate themselves whilst the 'Technicians' of yesterday didn't (well not often). The sad demise of mercury, lead, Bunsen burners, formalin, anything too hot, too cold, too explosive, too poisonous, etc. Having your tea in the Lab next to the specimens and processing TB specimens 'on the bench'. Would Histology have the techniques and stains it now has if harmful chemicals had not been experimented with? Will anything new be discovered by the HistoTech if all that we can use is 'safe' chemicals and procedures? I don't see the kids fiddling with things like we use to, no explosions, no fires and no injuries. Am I just an old reactionary waiting to be put out to pasture and ruminate on what was? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of sheila adey Sent: 17 November 2007 20:41 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hi Netters, We are trying to minimize possible embedding contaminations. What are other people doing to prevent contamination due to forceps etc. Thanks in advanceSheila Adey HT MLTPort Huron HospitalMichigan _________________________________________________________________ Are you ready for Windows Live Messenger Beta 8.5 ? Get the latest for free today! http://entertainment.sympatico.msn.ca/WindowsLiveMessenger__________________ _____________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pdunlop720 <@t> gmail.com Sun Nov 18 05:48:40 2007 From: pdunlop720 <@t> gmail.com (Patty Dunlop) Date: Sun Nov 18 05:48:45 2007 Subject: [Histonet] Histotechnician vs. Histotechnologist exam and resources Message-ID: <80ab7bc60711180348k40b18d89wd0cf0abaf6fc1f35@mail.gmail.com> Hello, I am preparing to apply to take either the histotechnician exam (HT) or the histotechnologist exam (HTL) and am not sure which one to take. I have a b.s. in biology, but did not take any histo courses. Would the HTL be too bold for me? I worked 3 years in a pathology lab working with animal tissue at Merck, so I do not have a wide knowledge of histology from my work experience. Also, what reading/study materials are recommended for someone who needs to learn about all the basics for the exam? Thanks! Patty From rjbuesa <@t> yahoo.com Sun Nov 18 07:51:51 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sun Nov 18 07:51:55 2007 Subject: [Histonet] Quick Decalcification In-Reply-To: <96566f7e0711172141q1b16306dy6ca69aebfc734351@mail.gmail.com> Message-ID: <690304.56047.qm@web61222.mail.yahoo.com> First, there is no such a thing as "quick decalcification" since the time required is determined by the type of hard part (usually bone) and its calcium contents. All decalcifiers in some way affect the soft tissue and what is precisely looked for is having little effect on the soft tissue. On the other hand, I think you could dissolve bone, and everything else, in a few imuntes if you put your bone in sulfuric acid; you will get a quick and total destruction of everything. So "quick decalcification" is incompatible with cellular detail. They are inversely proportionate (speed of decalcification and soft tissue detail). Ren? J. wael swelam wrote: Dear Histonet members; Good morning to all of you. Do you have a formula that can be used to achieve quick decalcification for bone & Dental hard tissues without affecting the quality of the related soft tissues. Your kind help is highly appreciated Yours, Wael Mohamed Swelam BDS (Egypt), Oral Pathology MSc (Egypt), Oral Pathology PhD (Japan), Ass. Prof. Oral Biomedical Science dept. Division of Oral Pathology, Faculty of Dentistry, King Faisal University, Dammam, Saudi Arabia, P.O: 1982/3441, http://myprofile.cos.com/wmswelam _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. From rjbuesa <@t> yahoo.com Sun Nov 18 07:58:31 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sun Nov 18 07:58:43 2007 Subject: [Histonet] Histotechnician vs. Histotechnologist exam and resources In-Reply-To: <80ab7bc60711180348k40b18d89wd0cf0abaf6fc1f35@mail.gmail.com> Message-ID: <184731.40280.qm@web61211.mail.yahoo.com> Your limited experience will be a handicap for either exam, but since you ha ve a BS I don't see why you should opt for the HT instead of the HTL exam. Go for the HTL, you will be equally challenged and the outcome will be better. Now, there are additional things in the HTL exam (like IHC) that perhaps you will find more challenging. As to reading, any good book will sufice as a reading source, but nothing will give you the day to day knowledge but practice. Ren? J. Patty Dunlop wrote: Hello, I am preparing to apply to take either the histotechnician exam (HT) or the histotechnologist exam (HTL) and am not sure which one to take. I have a b.s. in biology, but did not take any histo courses. Would the HTL be too bold for me? I worked 3 years in a pathology lab working with animal tissue at Merck, so I do not have a wide knowledge of histology from my work experience. Also, what reading/study materials are recommended for someone who needs to learn about all the basics for the exam? Thanks! Patty _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Never miss a thing. Make Yahoo your homepage. From dermpathsy <@t> gmail.com Sun Nov 18 09:01:26 2007 From: dermpathsy <@t> gmail.com (Sate Hamza) Date: Sun Nov 18 09:01:32 2007 Subject: [Histonet] MiTF on a Ventana Immunostainer Message-ID: <8854ff80711180701s3ae9307xb028f97bfe5f1fc4@mail.gmail.com> Hello, One of my colleagues posted the following inquiry on another forum. "Does anyone have a protocol for using MITF-1 on a Ventana immunostainer? Ventana doesn't have the antibody so they are not much help in either acquiring the antibody or helping us get started using it." I was wondering if anyone is using MITF on a Ventana Immunostainer and would be able to help. Many thanks in advance. Sate -- Sate Hamza, MD, FRCPC Dermatopathologist Winnipeg, Canada From contact <@t> excaliburpathology.com Sun Nov 18 10:12:38 2007 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Sun Nov 18 10:12:43 2007 Subject: [Histonet] no subject Message-ID: <651057.43222.qm@web50104.mail.re2.yahoo.com> Amen Kemlo! If we all are forced to "GO Green", we might as well turn out the lights and shut the doors. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com From rjbuesa <@t> yahoo.com Sun Nov 18 10:17:53 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sun Nov 18 10:17:59 2007 Subject: [Histonet] no subject In-Reply-To: <651057.43222.qm@web50104.mail.re2.yahoo.com> Message-ID: <325174.69314.qm@web61221.mail.yahoo.com> ...., and drink a large cup of hemlock! Ren? J. Paula Pierce wrote: Amen Kemlo! If we all are forced to "GO Green", we might as well turn out the lights and shut the doors. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From tkngflght <@t> yahoo.com Sun Nov 18 10:21:17 2007 From: tkngflght <@t> yahoo.com (Cheryl R. Kerry) Date: Sun Nov 18 10:21:52 2007 Subject: [Histonet] forceps and bunsen burners In-Reply-To: Message-ID: <037a01c829ff$0cc44dd0$6d01a8c0@CHERYLSLAPTOP> I really miss Bunsen burners! Now we use a lot of kimwipes. Kemlo--I recall the days of wiping your embedding center with a xylene soaked rag...me with a cuppa joe on my 'tome and others with cigarettes hanging out the sides of their mouths...and yes, the occasional lab fires that resulted!! And I really miss REAL light green SF yellowish! Cheryl -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kemlo Sent: Sunday, November 18, 2007 4:22 AM To: 'sheila adey'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] (no subject) In the good old days of Bunsen burners we used to fry the end of the forceps; nothing survived that and if it did it was readily identifiable. Alas the Bunsen burner has been consigned to the politically incorrect as the 'Scientists' of today would incinerate themselves whilst the 'Technicians' of yesterday didn't (well not often). The sad demise of mercury, lead, Bunsen burners, formalin, anything too hot, too cold, too explosive, too poisonous, etc. Having your tea in the Lab next to the specimens and processing TB specimens 'on the bench'. Would Histology have the techniques and stains it now has if harmful chemicals had not been experimented with? Will anything new be discovered by the HistoTech if all that we can use is 'safe' chemicals and procedures? I don't see the kids fiddling with things like we use to, no explosions, no fires and no injuries. Am I just an old reactionary waiting to be put out to pasture and ruminate on what was? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of sheila adey Sent: 17 November 2007 20:41 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hi Netters, We are trying to minimize possible embedding contaminations. What are other people doing to prevent contamination due to forceps etc. Thanks in advanceSheila Adey HT MLTPort Huron HospitalMichigan _________________________________________________________________ Are you ready for Windows Live Messenger Beta 8.5 ? Get the latest for free today! http://entertainment.sympatico.msn.ca/WindowsLiveMessenger__________________ _____________________________ Histonet mailing list Histonet@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 Sun Nov 18 12:18:04 2007 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Sun Nov 18 12:18:40 2007 Subject: [Histonet] Quick Decalcification In-Reply-To: <690304.56047.qm@web61222.mail.yahoo.com> References: <96566f7e0711172141q1b16306dy6ca69aebfc734351@mail.gmail.com> <690304.56047.qm@web61222.mail.yahoo.com> Message-ID: <000701c82a0f$5dd511e0$0202a8c0@ihctechq9h2qof> I agree with Rene, even some of the so called "rapid decal" reagents on the market can be detrimental to cellular morphology and connective tissue, especially if you are interested in doing IHC. That being said, I have seen some tissues (not bone) treated with strong formic acid to disinfect CJD which looked good morphologically by H&E. I did not test IHC on those samples. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 email pruegg@ihctech.net website www.ihctech.net IHC Resource Group www.ihcrg.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Sunday, November 18, 2007 6:52 AM To: wael swelam; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Quick Decalcification First, there is no such a thing as "quick decalcification" since the time required is determined by the type of hard part (usually bone) and its calcium contents. All decalcifiers in some way affect the soft tissue and what is precisely looked for is having little effect on the soft tissue. On the other hand, I think you could dissolve bone, and everything else, in a few imuntes if you put your bone in sulfuric acid; you will get a quick and total destruction of everything. So "quick decalcification" is incompatible with cellular detail. They are inversely proportionate (speed of decalcification and soft tissue detail). Ren? J. wael swelam wrote: Dear Histonet members; Good morning to all of you. Do you have a formula that can be used to achieve quick decalcification for bone & Dental hard tissues without affecting the quality of the related soft tissues. Your kind help is highly appreciated Yours, Wael Mohamed Swelam BDS (Egypt), Oral Pathology MSc (Egypt), Oral Pathology PhD (Japan), Ass. Prof. Oral Biomedical Science dept. Division of Oral Pathology, Faculty of Dentistry, King Faisal University, Dammam, Saudi Arabia, P.O: 1982/3441, http://myprofile.cos.com/wmswelam _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From contact <@t> excaliburpathology.com Sun Nov 18 12:42:26 2007 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Sun Nov 18 12:42:31 2007 Subject: [Histonet] forceps and bunsen burners Message-ID: <333065.27689.qm@web50106.mail.re2.yahoo.com> Hey! I never smoked while I was coverslipping! ;) There are several stains I miss. The dye content in some today sucks! Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com From Shirley_PHUA <@t> hsa.gov.sg Sun Nov 18 14:03:15 2007 From: Shirley_PHUA <@t> hsa.gov.sg (Shirley PHUA) Date: Sun Nov 18 14:05:19 2007 Subject: [Histonet] Shirley Phua is away on 19-23 November 2007. Message-ID: I will be out of the office from 19-11-2007 to 23-11-2007. I'll be away on 19-23 November 2007. I'll be back on 26 November 2007. Pathologists: I will process your requests when I return. If urgent, please forward your email to Henry_Kyaw@hsa.gov.sg From coleman_manufacturing <@t> yahoo.com Sun Nov 18 16:08:42 2007 From: coleman_manufacturing <@t> yahoo.com (Coleman Manufacturing) Date: Sun Nov 18 16:08:45 2007 Subject: [Histonet] UNSUBSCRIBE!!!!!!!!!!!!!!!!!!! Message-ID: <111864.77743.qm@web37603.mail.mud.yahoo.com> UNSUBSCRIBE!! This is the second email; PLEASE delete this email address from the list serve. From AnthonyH <@t> chw.edu.au Sun Nov 18 16:18:46 2007 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Sun Nov 18 16:20:03 2007 Subject: [Histonet] Minimum time processing Message-ID: Most important to ensure adequate fixation - as long as you can. At a minimum we fix 2-3mm diameter core bxs in 10% NBF for one hour and then 5 changes of alcohol 5 min each, xylene (x3) 5 min each and wax (x3) 5min each. 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: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Webb, Dorothy L Sent: Saturday, 17 November 2007 6:10 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Minimum time processing For those histology labs doing prostate needle biopsies.....I need some advice!! We process our pnbx on a separate program on a VIP 5 and these biospies are taken with a 20 gauge needle, very thin. What would be the minimum times you would recommend processing in the reagents? I am currently doing 10 minutes per station and I am seeing some very dry cores (they are usually the tiniest ones) that, of course, are staining very dark. I have a pathologist that feels we can do better, so needing my super histotechs to help me out once again!!! Thank you So much for help you always send via the histonet!! Dorothy Webb, HT (ASCP) Histology Technical Supervisor Regions Hospital, Pathology Department 640 Jackson Street, Saint Paul, MN 55101-2595 Phone: 651-254-2962 Fax: 651-254-2741 Regions Hospital is part of the HealthPartners family of care ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************* 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 note 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 cfarish <@t> csu.edu.au Sun Nov 18 16:23:46 2007 From: cfarish <@t> csu.edu.au (Farish, Craig) Date: Sun Nov 18 16:23:56 2007 Subject: [Histonet] R.E. no subject (embedding contamination) Message-ID: <79A000B60AFE8045BA2581119DFEC44402A251E1@ESWW01.CSUMain.csu.edu.au> You can also have the grooves filed from the tip of your embedding forceps (or purchase forceps without grooves if possible). They still perform the same function but can't carry over contamination as easily. We're looking at the enforced removal of Picric Acid from histo labs here. Where's the fun in non-exploding reagents? Craig (Joe) Farish Technical Officer School of Agricultural and Veterinary Sciences Charles Sturt University Wagga Wagga NSW 2678 Australia ' I don't want to achieve immortality through my work, I want to achieve immortality through not dying' - Woody Allen From tjey <@t> hotmail.com Sun Nov 18 21:51:53 2007 From: tjey <@t> hotmail.com (Tanya Ewing-Finchem) Date: Sun Nov 18 21:51:58 2007 Subject: [Histonet] MiTF on a Ventana Immunostainer In-Reply-To: References: Message-ID: Have you asked your Ventana Sales Rep about help with the situation? ;-) It has been my experience that Ventana has an applications lab that can help you with a 3rd party Ab. work up. > > Message: 11> Date: Sun, 18 Nov 2007 09:01:26 -0600> From: "Sate Hamza" > Subject: [Histonet] MiTF on a Ventana Immunostainer> To: histonet@lists.utsouthwestern.edu> Message-ID:> <8854ff80711180701s3ae9307xb028f97bfe5f1fc4@mail.gmail.com>> Content-Type: text/plain; charset=ISO-8859-1> > Hello,> > One of my colleagues posted the following inquiry on another forum.> > "Does anyone have a protocol for using MITF-1 on a Ventana immunostainer?> Ventana doesn't have the antibody so they are not much help in either> acquiring the antibody or helping us get started using it."> > I was wondering if anyone is using MITF on a Ventana Immunostainer and would> be able to help.> > Many thanks in advance.> > Sate> > > -- > Sate Hamza, MD, FRCPC> Dermatopathologist> Winnipeg, Canada> > From dermpathsy <@t> gmail.com Sun Nov 18 22:39:39 2007 From: dermpathsy <@t> gmail.com (Sate Hamza) Date: Sun Nov 18 22:39:44 2007 Subject: [Histonet] MiTF on a Ventana Immunostainer In-Reply-To: References: Message-ID: <8854ff80711182039yee9e8c6l283a9e2b0e972064@mail.gmail.com> It sounds to me from my colleague's comment that she tried contacting Ventana regarding this, but that they were not of much help. Thanks. Sate On 11/18/07, Tanya Ewing-Finchem wrote: > > > Have you asked your Ventana Sales Rep about help with the situation? ;-) > It has been my experience that Ventana has an applications lab that can > help you with a 3rd party Ab. work up. > > [--] "Does anyone have a protocol for using MITF-1 on a Ventana > immunostainer? Ventana doesn't have the antibody so they are not much help > in either acquiring the antibody or helping us get started using it."> >[--] -- Sate Hamza, MD, FRCPC Dermatopathologist Winnipeg, Canada From ree3 <@t> leicester.ac.uk Mon Nov 19 04:41:39 2007 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Mon Nov 19 04:41:46 2007 Subject: [Histonet] no subject In-Reply-To: <651057.43222.qm@web50104.mail.re2.yahoo.com> References: <651057.43222.qm@web50104.mail.re2.yahoo.com> Message-ID: You/we might not be given any choice in the future!. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Pierce Sent: 18 November 2007 16:13 To: Histonet Subject: [Histonet] no subject Amen Kemlo! If we all are forced to "GO Green", we might as well turn out the lights and shut the doors. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> waht.swest.nhs.uk Mon Nov 19 04:59:59 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Nov 19 05:00:07 2007 Subject: [Histonet] no subject Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F1B2@wahtntex2.waht.swest.nhs.uk> Amen Kemlo! If we all are forced to "GO Green", we might as well turn out the lights and shut the doors. Paula Pierce, HTL(ASCP)HT Whilst your comments are common sense, I fear the lights are being extinguished as we speak. Technicians/ Scientists who don't experiment lest they hurt themselves or someone else. It all went wrong when they stopped Lab Staff making cocktails out of ethyl alcohol at Christmas; I know some got ethyl and methyl mixed up and went blind but they soon learnt about the formulae of organic compounds. We used to have cyanide and strychnine in the Lab and oddly I never felt the inclination to ingest, odd that. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From oshel1pe <@t> cmich.edu Mon Nov 19 07:35:46 2007 From: oshel1pe <@t> cmich.edu (Philip Oshel) Date: Mon Nov 19 07:35:55 2007 Subject: [Histonet] (no subject) In-Reply-To: References: Message-ID: Ah, I no longer feel so alone. I recall using bunsen burners in junior high (that ought to date me -- "middle school" now), high school, as a college freshman and so on. Never a problem, no burned-down schools, no explosions, no kids on fire. (Mind, my brother did make nitrate contact explosives with his chemistry set, so I never got to have one.) Is common sense so dead that even experienced lab workers must be protected from themselves? I've never understood the anti-bunsen paranoia. Someone should write a history of the bunsen burner: a modest little device, but where would chemistry, microbiology, and science in general be without it? Phil >In the good old days of Bunsen burners we used to fry the end of the >forceps; nothing survived that and if it did it was readily identifiable. >Alas the Bunsen burner has been consigned to the politically incorrect as >the 'Scientists' of today would incinerate themselves whilst the >'Technicians' of yesterday didn't (well not often). > >The sad demise of mercury, lead, Bunsen burners, formalin, anything too hot, >too cold, too explosive, too poisonous, etc. Having your tea in the Lab next >to the specimens and processing TB specimens 'on the bench'. > >Would Histology have the techniques and stains it now has if harmful >chemicals had not been experimented with? Will anything new be discovered by >the HistoTech if all that we can use is 'safe' chemicals and procedures? I >don't see the kids fiddling with things like we use to, no explosions, no >fires and no injuries. Am I just an old reactionary waiting to be put out to >pasture and ruminate on what was? > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of sheila adey >Sent: 17 November 2007 20:41 >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] (no subject) > > >Hi Netters, > >We are trying to minimize possible embedding contaminations. What are other >people doing to prevent contamination due to forceps etc. > >Thanks in advanceSheila Adey HT MLTPort Huron HospitalMichigan From doug <@t> ppspath.com Mon Nov 19 07:54:03 2007 From: doug <@t> ppspath.com (Douglas D Deltour) Date: Mon Nov 19 07:54:24 2007 Subject: SPAM-LOW: [Histonet] MiTF on a Ventana Immunostainer In-Reply-To: <8854ff80711180701s3ae9307xb028f97bfe5f1fc4@mail.gmail.com> Message-ID: We just started using the MITF-1 antibody on the XT. I got it validated and we were running it for about 2 months and then all of a sudden it stopped working, but that is another story. Which company did your colleague get the antibody from? I used the same protocol that I used for the MART-1 for validation. This was working great for the MITF until..... Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sate Hamza Sent: Sunday, November 18, 2007 10:01 AM To: histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: [Histonet] MiTF on a Ventana Immunostainer Hello, One of my colleagues posted the following inquiry on another forum. "Does anyone have a protocol for using MITF-1 on a Ventana immunostainer? Ventana doesn't have the antibody so they are not much help in either acquiring the antibody or helping us get started using it." I was wondering if anyone is using MITF on a Ventana Immunostainer and would be able to help. Many thanks in advance. Sate -- Sate Hamza, MD, FRCPC Dermatopathologist Winnipeg, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From doug <@t> ppspath.com Mon Nov 19 08:08:34 2007 From: doug <@t> ppspath.com (Douglas D Deltour) Date: Mon Nov 19 08:08:56 2007 Subject: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer In-Reply-To: <8854ff80711182039yee9e8c6l283a9e2b0e972064@mail.gmail.com> Message-ID: Unfortunately Ventana will not give you much help unless it is a Ventana antibody. I can understand if they offered the antibody and you were using a competitor but they do not offer the MITF-1. If the majority of users are requesting help with the same types of third party antibodies(not offered by Ventana) then it may behoove Ventana (for the full customer service experience) to offer some suggestion with these antibodies (creating another job for someone (cough cough)). We still are paying for the prep-kit which may cost more than some antibodies. Just a thought. Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sate Hamza Sent: Sunday, November 18, 2007 11:40 PM To: Histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer It sounds to me from my colleague's comment that she tried contacting Ventana regarding this, but that they were not of much help. Thanks. Sate On 11/18/07, Tanya Ewing-Finchem wrote: > > > Have you asked your Ventana Sales Rep about help with the situation? ;-) > It has been my experience that Ventana has an applications lab that can > help you with a 3rd party Ab. work up. > > [--] "Does anyone have a protocol for using MITF-1 on a Ventana > immunostainer? Ventana doesn't have the antibody so they are not much help > in either acquiring the antibody or helping us get started using it."> >[--] -- Sate Hamza, MD, FRCPC Dermatopathologist Winnipeg, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From contact <@t> excaliburpathology.com Mon Nov 19 08:36:43 2007 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Mon Nov 19 08:36:48 2007 Subject: [Histonet] no subject Message-ID: <119713.97956.qm@web50111.mail.re2.yahoo.com> So when are we going to say, "we're mad as hell, & we're not gonna take it anymore!" ? Why bother teaching lab safety, if the potential lab of the future is so regulated nothing is harmful? My high school chemistry teacher took the entire class outside one day and he placed a chunk of pure sodium under an empty paint can. (of course this was decades ago) When enough moisture had reacted with it, that paint can was blown higher than the 2 story building. It was so cool, but we also learned what can happen. If that happened today, some parent would sue for endangering their child. We use formalin because it works! We use xylene because it works! We use ___________ because it works! If you stand for nothing, you will fall for anything! Paula From talulahgosh <@t> gmail.com Mon Nov 19 08:44:19 2007 From: talulahgosh <@t> gmail.com (Emily Sours) Date: Mon Nov 19 08:44:24 2007 Subject: [Histonet] forceps and bunsen burners In-Reply-To: <037a01c829ff$0cc44dd0$6d01a8c0@CHERYLSLAPTOP> References: <037a01c829ff$0cc44dd0$6d01a8c0@CHERYLSLAPTOP> Message-ID: We still use bunsen burners and alcohol flames to sterilize forceps. We also make our own tungsten needles from tungsten wire and capillary tubes, burning them to a point in a blast burner. We like our antiquated techniques. Emily -- crow tom mike and joel trapped in space the endless void we've got movie sign! mst3k haiku brought to you by joe wiencis From talulahgosh <@t> gmail.com Mon Nov 19 08:45:59 2007 From: talulahgosh <@t> gmail.com (Emily Sours) Date: Mon Nov 19 08:46:08 2007 Subject: [Histonet] no subject In-Reply-To: <119713.97956.qm@web50111.mail.re2.yahoo.com> References: <119713.97956.qm@web50111.mail.re2.yahoo.com> Message-ID: Union! Union! Union! On Nov 19, 2007 9:36 AM, Paula Pierce wrote: > So when are we going to say, "we're mad as hell, & we're not gonna take it > anymore!" ? > > Why bother teaching lab safety, if the potential lab of the future is so > regulated nothing is harmful? > > My high school chemistry teacher took the entire class outside one day and > he placed a chunk of pure sodium under an empty paint can. (of course this > was decades ago) When enough moisture had reacted with it, that paint can > was blown higher than the 2 story building. It was so cool, but we also > learned what can happen. If that happened today, some parent would sue for > endangering their child. > > We use formalin because it works! > We use xylene because it works! > We use ___________ because it works! > > If you stand for nothing, you will fall for anything! > > Paula > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- crow tom mike and joel trapped in space the endless void we've got movie sign! mst3k haiku brought to you by joe wiencis From doug <@t> ppspath.com Mon Nov 19 09:01:52 2007 From: doug <@t> ppspath.com (Douglas D Deltour) Date: Mon Nov 19 09:02:24 2007 Subject: SPAM-LOW: Re: [Histonet] no subject In-Reply-To: Message-ID: I honor of this thread I declare Thursday November 22nd National Histology Strike Day! We will not report to work on this day to show our dissatisfaction. Who is with me? Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Emily Sours Sent: Monday, November 19, 2007 9:46 AM To: histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: Re: [Histonet] no subject Union! Union! Union! On Nov 19, 2007 9:36 AM, Paula Pierce wrote: > So when are we going to say, "we're mad as hell, & we're not gonna take it > anymore!" ? > > Why bother teaching lab safety, if the potential lab of the future is so > regulated nothing is harmful? > > My high school chemistry teacher took the entire class outside one day and > he placed a chunk of pure sodium under an empty paint can. (of course this > was decades ago) When enough moisture had reacted with it, that paint can > was blown higher than the 2 story building. It was so cool, but we also > learned what can happen. If that happened today, some parent would sue for > endangering their child. > > We use formalin because it works! > We use xylene because it works! > We use ___________ because it works! > > If you stand for nothing, you will fall for anything! > > Paula > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- crow tom mike and joel trapped in space the endless void we've got movie sign! mst3k haiku brought to you by joe wiencis _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kgrobert <@t> rci.rutgers.edu Mon Nov 19 09:10:10 2007 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Mon Nov 19 09:02:53 2007 Subject: [Histonet] Overdoing Safety/bunsen burners In-Reply-To: References: Message-ID: <4741A752.6040000@rci.rutgers.edu> Most of the safety people here are a bit like that....paranoid, that is; but there was one woman a few years ago who actually did say to me, "you're grown up, you know what you're doing." She was originally from Russia and apparently had chosen not to swallow the "protect people from themselves and everything" line. I think she is still with us, just on a different campus. I always think it's funny when the safety people come around here, because we're part of the Pharmacology & Toxicology dept...who would know better than us what's dangerous and what isn't? :o) Kathleen Philip Oshel wrote: > Ah, I no longer feel so alone. I recall using bunsen burners in junior > high (that ought to date me -- "middle school" now), high school, as a > college freshman and so on. Never a problem, no burned-down schools, > no explosions, no kids on fire. (Mind, my brother did make nitrate > contact explosives with his chemistry set, so I never got to have > one.) Is common sense so dead that even experienced lab workers must > be protected from themselves? > I've never understood the anti-bunsen paranoia. Someone should write a > history of the bunsen burner: a modest little device, but where would > chemistry, microbiology, and science in general be without it? > > Phil > >> In the good old days of Bunsen burners we used to fry the end of the >> forceps; nothing survived that and if it did it was readily >> identifiable. >> Alas the Bunsen burner has been consigned to the politically >> incorrect as >> the 'Scientists' of today would incinerate themselves whilst the >> 'Technicians' of yesterday didn't (well not often). >> >> The sad demise of mercury, lead, Bunsen burners, formalin, anything >> too hot, >> too cold, too explosive, too poisonous, etc. Having your tea in the >> Lab next >> to the specimens and processing TB specimens 'on the bench'. >> >> Would Histology have the techniques and stains it now has if harmful >> chemicals had not been experimented with? Will anything new be >> discovered by >> the HistoTech if all that we can use is 'safe' chemicals and >> procedures? I >> don't see the kids fiddling with things like we use to, no >> explosions, no >> fires and no injuries. Am I just an old reactionary waiting to be put >> out to >> pasture and ruminate on what was? >> >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >> sheila adey >> Sent: 17 November 2007 20:41 >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] (no subject) >> >> >> Hi Netters, >> >> We are trying to minimize possible embedding contaminations. What are >> other >> people doing to prevent contamination due to forceps etc. >> >> Thanks in advanceSheila Adey HT MLTPort Huron HospitalMichigan > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> waht.swest.nhs.uk Mon Nov 19 09:09:30 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Nov 19 09:09:35 2007 Subject: SPAM-LOW: Re: [Histonet] no subject Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F1C8@wahtntex2.waht.swest.nhs.uk> I honor of this thread I declare Thursday November 22nd National Histology Strike Day! We will not report to work on this day to show our dissatisfaction. Who is with me? Douglas Where are you? Can I get you on my Sat Nav? What's your postcode? Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From lblazek <@t> digestivespecialists.com Mon Nov 19 09:17:17 2007 From: lblazek <@t> digestivespecialists.com (Blazek, Linda) Date: Mon Nov 19 09:13:36 2007 Subject: SPAM-LOW: Re: [Histonet] no subject In-Reply-To: References: Message-ID: <1F937FB30BDB7C4A9F39F83FEA8D379F3E4D5A@bruexchange1.digestivespecialists.com> I am! Linda Blazek HT (ASCP) Manager/Supervisor GI Pathology of Dayton 7415 Brandt Pike Huber Heights, OH 45424 Phone: (937) 293-4424 ext 7118 Email: lblazek@digestivespecialists.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Douglas D Deltour Sent: Monday, November 19, 2007 10:02 AM To: 'Emily Sours'; histonet@lists.utsouthwestern.edu Subject: RE: SPAM-LOW: Re: [Histonet] no subject I honor of this thread I declare Thursday November 22nd National Histology Strike Day! We will not report to work on this day to show our dissatisfaction. Who is with me? Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Emily Sours Sent: Monday, November 19, 2007 9:46 AM To: histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: Re: [Histonet] no subject Union! Union! Union! On Nov 19, 2007 9:36 AM, Paula Pierce wrote: > So when are we going to say, "we're mad as hell, & we're not gonna take it > anymore!" ? > > Why bother teaching lab safety, if the potential lab of the future is so > regulated nothing is harmful? > > My high school chemistry teacher took the entire class outside one day and > he placed a chunk of pure sodium under an empty paint can. (of course this > was decades ago) When enough moisture had reacted with it, that paint can > was blown higher than the 2 story building. It was so cool, but we also > learned what can happen. If that happened today, some parent would sue for > endangering their child. > > We use formalin because it works! > We use xylene because it works! > We use ___________ because it works! > > If you stand for nothing, you will fall for anything! > > Paula > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- crow tom mike and joel trapped in space the endless void we've got movie sign! mst3k haiku brought to you by joe wiencis _______________________________________________ Histonet mailing list Histonet@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 talulahgosh <@t> gmail.com Mon Nov 19 09:38:01 2007 From: talulahgosh <@t> gmail.com (Emily Sours) Date: Mon Nov 19 09:38:06 2007 Subject: [Histonet] no subject Message-ID: I'm totally with you! Be sure to let everyone know we're on strike. Emily -- crow tom mike and joel trapped in space the endless void we've got movie sign! mst3k haiku brought to you by joe wiencis From slappycraw <@t> yahoo.com Mon Nov 19 09:42:29 2007 From: slappycraw <@t> yahoo.com (Larry Woody) Date: Mon Nov 19 09:42:35 2007 Subject: [Histonet] Overdoing Safety/bunsen burners In-Reply-To: <4741A752.6040000@rci.rutgers.edu> Message-ID: <965432.80991.qm@web53611.mail.re2.yahoo.com> It's the Ralph Nader Syndrom, let's make the world into a nurf ball so no one ever gets hurt. My take on it is this: When safety is out of control, we are all in danger! I know they are just trying to do their jobs and protect companies from the lawsuits but you have to admit there are some safety and security people that take things too far. I once worked at a hospital where the head of security wanted to build guard towers on all four corners of the hospital property and he almost succeeded! Kathleen Roberts wrote: Most of the safety people here are a bit like that....paranoid, that is; but there was one woman a few years ago who actually did say to me, "you're grown up, you know what you're doing." She was originally from Russia and apparently had chosen not to swallow the "protect people from themselves and everything" line. I think she is still with us, just on a different campus. I always think it's funny when the safety people come around here, because we're part of the Pharmacology & Toxicology dept...who would know better than us what's dangerous and what isn't? :o) Kathleen Philip Oshel wrote: > Ah, I no longer feel so alone. I recall using bunsen burners in junior > high (that ought to date me -- "middle school" now), high school, as a > college freshman and so on. Never a problem, no burned-down schools, > no explosions, no kids on fire. (Mind, my brother did make nitrate > contact explosives with his chemistry set, so I never got to have > one.) Is common sense so dead that even experienced lab workers must > be protected from themselves? > I've never understood the anti-bunsen paranoia. Someone should write a > history of the bunsen burner: a modest little device, but where would > chemistry, microbiology, and science in general be without it? > > Phil > >> In the good old days of Bunsen burners we used to fry the end of the >> forceps; nothing survived that and if it did it was readily >> identifiable. >> Alas the Bunsen burner has been consigned to the politically >> incorrect as >> the 'Scientists' of today would incinerate themselves whilst the >> 'Technicians' of yesterday didn't (well not often). >> >> The sad demise of mercury, lead, Bunsen burners, formalin, anything >> too hot, >> too cold, too explosive, too poisonous, etc. Having your tea in the >> Lab next >> to the specimens and processing TB specimens 'on the bench'. >> >> Would Histology have the techniques and stains it now has if harmful >> chemicals had not been experimented with? Will anything new be >> discovered by >> the HistoTech if all that we can use is 'safe' chemicals and >> procedures? I >> don't see the kids fiddling with things like we use to, no >> explosions, no >> fires and no injuries. Am I just an old reactionary waiting to be put >> out to >> pasture and ruminate on what was? >> >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >> sheila adey >> Sent: 17 November 2007 20:41 >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] (no subject) >> >> >> Hi Netters, >> >> We are trying to minimize possible embedding contaminations. What are >> other >> people doing to prevent contamination due to forceps etc. >> >> Thanks in advanceSheila Adey HT MLTPort Huron HospitalMichigan > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Larry A. Woody Amgen Seattle, Wa. --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. From relia1 <@t> earthlink.net Mon Nov 19 09:57:29 2007 From: relia1 <@t> earthlink.net (Pam Barker) Date: Mon Nov 19 09:57:32 2007 Subject: [Histonet] Histo Tech Needed in Los Angeles. Can you help? Message-ID: Hello Histonetters, I hope everybody is enjoying the beautiful autumn weather. I am currently working with a client in the Los Angeles area that is in need of junior histo tech or Mohs Tech this is a large dermatopathology practice and you will have the opportunity to learn Mohs. This is a permanent full time dayshift position. The client offers a great environment, a great crew to work with and competitive salary and benefits. Experienced Techs and New Grads are welcome to apply. My question is do you know of anyone who might be interested in this position? If you think you or someone you know might be interested please contact me. I can be reached at 866-607-3542 or relia1@earthlink.net Thanks-Pam 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 www.myspace.com/pamatrelia From alaskagirl1950 <@t> yahoo.com Mon Nov 19 10:23:03 2007 From: alaskagirl1950 <@t> yahoo.com (Patricia Adams) Date: Mon Nov 19 10:23:07 2007 Subject: [Histonet] Histology Strike! Message-ID: <875640.56798.qm@web52511.mail.re2.yahoo.com> Both Elma, and I plan to carry the strike over through Friday. No one will be here to notice, bummer. Patricia Adams ----- Fight back spam! Download the Blue Frog. http://www.bluesecurity.com/register/s?user=YWxhc2thZ2lybDE5NTA%3D ____________________________________________________________________________________ Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. http://mobile.yahoo.com/sports;_ylt=At9_qDKvtAbMuh1G1SQtBI7ntAcJ From ree3 <@t> leicester.ac.uk Mon Nov 19 10:23:02 2007 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Mon Nov 19 10:23:12 2007 Subject: [Histonet] no subject In-Reply-To: References: Message-ID: But, who will notice??. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Emily Sours Sent: 19 November 2007 15:38 To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] no subject I'm totally with you! Be sure to let everyone know we're on strike. Emily -- crow tom mike and joel trapped in space the endless void we've got movie sign! mst3k haiku brought to you by joe wiencis _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PIXLEYSK <@t> UCMAIL.UC.EDU Mon Nov 19 10:40:18 2007 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Mon Nov 19 10:40:24 2007 Subject: [Histonet] RE: Rapid decalcifiers In-Reply-To: <200711191624.HLX68760@mprelay2.uc.edu> Message-ID: <771F0CF37F5F7E44884D8022D2A3EAB41EB15E@ucmail6.ad.uc.edu> RE: Decalcifying: We found that when we used a Rapid decalcifier (RDO) with our bony tissue, we could not get BrdU immunostaining to work. The BrdU stain worked with EDTA decalcification, but that takes a very long time (> 1 week). Then, we found that we could get great staining with 10% Formic acid, which takes only 3-4 days. We put in an ion exchange resin also to soak up the ions and keep it in a fume hood, stirring, with the samples suspended from strings, in biopsy bags. Ion exchange resin: Rexyn 101 H ion exchange resin Fisher # 231-500. Formula: Make 6-10 liters of 10% formic acid (10% from the stock, which is actually 90%) Add 500 g of resin Sarah Pixley From dermpathsy <@t> gmail.com Mon Nov 19 11:22:59 2007 From: dermpathsy <@t> gmail.com (Sate Hamza) Date: Mon Nov 19 11:23:07 2007 Subject: [Histonet] MiTF on a Ventana Immunostainer Message-ID: <8854ff80711190922y18de3ca0u1219e842605867e6@mail.gmail.com> Thanks to all who have responded to my message. I asked my colleague and she said that she has not ordered any antibody yet. She said she wanted to see if she can get a protocol first before she orders the antibody. She was wondering what the source of your antibody was .. Thanks again .. Sate On 11/19/07, Douglas D Deltour wrote: > > We just started using the MITF-1 antibody on the XT. I got it validated > and > we were running it for about 2 months and then all of a sudden it stopped > working, but that is another story. Which company did your colleague get > the > antibody from? I used the same protocol that I used for the MART-1 for > validation. This was working great for the MITF until..... > > [---] > > > > -- Sate Hamza, MD, FRCPC Dermatopathologist Winnipeg, Canada From ROrr <@t> enh.org Mon Nov 19 11:24:11 2007 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Mon Nov 19 11:24:21 2007 Subject: [Histonet] RE: Histonet Digest, Vol 48, Issue 17 Message-ID: Hi Leslie, Our lab is running the Auto TEC from Sakura and it's working wonderfully. Let me know if you have any questions, we'll be glad to help. Becky Becky Orr CLA,HT(ASCP)QIHC Anatomic Pathology Evanston Northwestern Healthcare 847-570-2771 Message: 13 Date: Tue, 13 Nov 2007 11:01:02 -0500 From: "Lesley Bechtold" Subject: [Histonet] automated paraffin embedding systems To: "Histology Network" Message-ID: <20071113110102345.00000002152@spikey> Content-Type: text/plain; charset=us-ascii Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, November 13, 2007 10:36 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 48, 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. Getting a Tissue Processor (Jennifer Sipes) 2. non pathologist grossing vs processing continued (Vancleave, Vince) 3. RE: non pathologist grossing vs processing continued (Charles.Embrey) 4. Susie Hargrove is out of the office. (SHargrove@urhcs.org) 5. RE: deplasticizing PMMA sections for staining (Michelle McDonald) 6. MyoD1 (San Tin) 7. RE: TGIF (Farish, Craig) 8. Claudin-1 as a prognostic factor in colorectal cancer (K M) 9. staining of negative controls with IHC (MaryAnn Dixon) 10. Re: staining of negative controls with IHC (Rene J Buesa) 11. Re: staining of negative controls with IHC (AGrobe2555@aol.com) 12. RE: staining of negative controls with IHC (Tarango, Mark) 13. automated paraffin embedding systems (Lesley Bechtold) 14. Re: automated paraffin embedding systems (Rene J Buesa) 15. RE: automated paraffin embedding systems (Mike Pence) 16. RE: automated paraffin embedding systems (Lesley Bechtold) 17. Antibody cocktails (Christine I. Braaten) 18. RE: staining of negative controls with IHC (FU,DONGTAO) ---------------------------------------------------------------------- Message: 1 Date: Mon, 12 Nov 2007 10:14:18 -0800 (PST) From: Jennifer Sipes Subject: [Histonet] Getting a Tissue Processor To: histonet@lists.utsouthwestern.edu Message-ID: <750683.40287.qm@web62408.mail.re1.yahoo.com> Content-Type: text/plain; charset=us-ascii My lab is FINALLY getting a Shandon Excelsior Tissue Processor. I was wondering if anyone had any good protocols for the machine that would be great for mouse tissue...mainly the GI trac and liver and kidneys. Thanks for all of your help! Jen __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 2 Date: Mon, 12 Nov 2007 13:54:54 -0600 From: "Vancleave, Vince" Subject: [Histonet] non pathologist grossing vs processing continued To: Message-ID: <740F77F6594C20458BDC2A4A500D8D41501642@EMAIL.hhs.hendrickhealth.org> Content-Type: text/plain; charset="iso-8859-1" From: Vancleave, Vince Sent: Wed 11/7/2007 2:39 PM To: histonet@lists.utsouthwestern.edu Cc: Webster, John A. M.D. Subject: Still not sure about this low complexity or not "The thing you have to remember, Vince, is that CLIA and CAP are two different, completely separate entities. CLIA is government controlled and CAP is private. Whereas CAP recently decided to separate grossing into two different complexities, CLIA has not. Bottom line...if you are CLIA licensed you MUST follow the CLIA standard. If not, then you can do whatever you like. Charles Embrey Jr., PA(ASCP)" Charles makes a great point but: I'm just not sure I'm sold on this yet, b/c also in the CLIA regs it states in sec. 493.559 about exemption from CLIA, via using a private accreditation agency (which is what CAP is), and states clearly that this agency can only be deemed acceptable if their requirements for the laboratory are equivalent or more stringent and the laboratory would meet CLIA requirements if the laboratory had not been granted deemed status. Ya'll are right, this is a mess! So, wouldn't this essentially mean that according to CLIA, since CAP is an acceptable private agency, they are giving the ok for CAP's decision on this? Wouldn't CAP be unallowed to do what they do if it was less stringent than CLIA? I also find nothing in the entire CLIA 88' that indicates that this would not be ok. In fact, CLIA has a criteria for categorization of complexity: http://www.fda.gov/cdrh/clia/categorization.html . And in their database of tests that they have said has to absolutely be a certain way, I find no reference to tissue description, gross examination, tissue examination, or processing. Have I stumped anyone yet, or is there some more things I am missing? I really appreciate all ya'lls willingness to re-visit this issue again. I guess I am a stuborn monkey! Thanks, Vince Van Cleave, BS, PA(ASCP), HTL, HT Laboratory Manager and Pathologist Assistant (325)670-6528 Clinical Pathology Associates Abilene, TX PRIVILEGED AND CONFIDENTIAL NOTICE: The information contained in this e-mail may be confidential and/or privileged. This e-mail is intended to be reviewed initially by only the individual named above. If the reader of this e-mail is not the intended recipient or a representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail, or the information contained herein, is prohibited. If you have received this e-mail in error, notify the above sender, send the message back and then delete it. Thank you for your assistance. Hendrick Health System 1900 Pine St. Abilene, Texas 79601 - 325.670.2000 79601 - 325.670.2000 ------------------------------ Message: 3 Date: Mon, 12 Nov 2007 14:28:36 -0600 From: "Charles.Embrey" Subject: RE: [Histonet] non pathologist grossing vs processing continued To: "Vancleave, Vince" , Message-ID: <44780C571F28624DBB446DE55C4D733A1FE56A@EXCHANGEBE1.carle.com> Content-Type: text/plain; charset="us-ascii" Hi Vince, Sorry to take so long in getting back to you but things here have been very busy. By CLIA's guidelines it sounds as though CAP has removed themselves as an "acceptable" private accreditation when they loosened the restrictions on grossing. CLIA's database may not list grossing but CLIA did publish a guideline to clarify their position: This is the CLIA '88 guideline for high complexity testing personnel- CLIA '88 493.1489 "On or before April 24 1995 (I) be a high school graduate or equivalent; and (b) have documentation of training appropriate for the test performed before analyzing patient specimens"...After that date it requires an associate degree in a biological or chemical science or medical laboratory technology -or- qualify as a medical technologist with a bachelor's degree from an accredited institution -or- earned a bachelor's degree in a chemical, physical, biologic or clinical laboratory science. A few years ago CLIA put out this interpretive guideline, (printed below), to clear up the question of grossing. As you see, it references 493.1498 (the above "high complexity guideline"). They basically sum up that if you examine/describe the tissue for record noting color weight or measurement then it is grossing and falls under 493.1498. Tissue processing as CAP calls it involves noting size and other characteristics and qualifies as grossing under CLIA. The new federal register CLIA interpretive guidelines appendix C subpart M effective April 24, 2003 states in section 493.1461(e) "In the case if gross examinations, the technical supervisor may delegate to individuals qualified under 493.1498 the responsibility for the physical examination/description, including color, weight, measurement, and other characteristics of the tissue; or other mechanical procedures for which a specific written protocol has been developed." I hope this helps. I don't know which side will budge first but I have heard whisperings of CLIA doing something in the near future but what ever it is it will probably only muddy the water further. All I can say is that CLIA is what happens when a government bureaucracy gets involved with healthcare. Charles Embrey PA(ASCP) Histology Manager Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vancleave, Vince Sent: Monday, November 12, 2007 1:55 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] non pathologist grossing vs processing continued From: Vancleave, Vince Sent: Wed 11/7/2007 2:39 PM To: histonet@lists.utsouthwestern.edu Cc: Webster, John A. M.D. Subject: Still not sure about this low complexity or not "The thing you have to remember, Vince, is that CLIA and CAP are two different, completely separate entities. CLIA is government controlled and CAP is private. Whereas CAP recently decided to separate grossing into two different complexities, CLIA has not. Bottom line...if you are CLIA licensed you MUST follow the CLIA standard. If not, then you can do whatever you like. Charles Embrey Jr., PA(ASCP)" Charles makes a great point but: I'm just not sure I'm sold on this yet, b/c also in the CLIA regs it states in sec. 493.559 about exemption from CLIA, via using a private accreditation agency (which is what CAP is), and states clearly that this agency can only be deemed acceptable if their requirements for the laboratory are equivalent or more stringent and the laboratory would meet CLIA requirements if the laboratory had not been granted deemed status. Ya'll are right, this is a mess! So, wouldn't this essentially mean that according to CLIA, since CAP is an acceptable private agency, they are giving the ok for CAP's decision on this? Wouldn't CAP be unallowed to do what they do if it was less stringent than CLIA? I also find nothing in the entire CLIA 88' that indicates that this would not be ok. In fact, CLIA has a criteria for categorization of complexity: http://www.fda.gov/cdrh/clia/categorization.html . And in their database of tests that they have said has to absolutely be a certain way, I find no reference to tissue description, gross examination, tissue examination, or processing. Have I stumped anyone yet, or is there some more things I am missing? I really appreciate all ya'lls willingness to re-visit this issue again. I guess I am a stuborn monkey! Thanks, Vince Van Cleave, BS, PA(ASCP), HTL, HT Laboratory Manager and Pathologist Assistant (325)670-6528 Clinical Pathology Associates Abilene, TX PRIVILEGED AND CONFIDENTIAL NOTICE: The information contained in this e-mail may be confidential and/or privileged. This e-mail is intended to be reviewed initially by only the individual named above. If the reader of this e-mail is not the intended recipient or a representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail, or the information contained herein, is prohibited. If you have received this e-mail in error, notify the above sender, send the message back and then delete it. Thank you for your assistance. Hendrick Health System 1900 Pine St. Abilene, Texas 79601 - 325.670.2000 79601 - 325.670.2000 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 12 Nov 2007 16:52:19 -0600 From: SHargrove@urhcs.org Subject: [Histonet] Susie Hargrove is out of the office. To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII I will be out of the office starting 11/12/2007 and will not return until 11/19/2007. I will respond to your message when I return. ------------------------------ Message: 5 Date: Tue, 13 Nov 2007 10:27:51 +1100 From: "Michelle McDonald" Subject: RE: [Histonet] deplasticizing PMMA sections for staining To: "Monfils, Paul" , Message-ID: <47BD6E7614A693499453835D47E36F7003882B4F@hedwig.nch.kids> Content-Type: text/plain; charset="us-ascii" We actually use Acetone to deplasticise MMA sections. Only takes 2 changes for 10 mins each. Michelle Dr Michelle McDonald B.MedSci, PhD Research Officer Orthopaedic Research Dept. The Children's Hospital Westmead. Westmead NSW 2145 Australia Ph. +612 9845 1451 Fax. +612 9845 3078 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Tuesday, 6 November 2007 6:22 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] deplasticizing PMMA sections for staining In my experience, xylene is a rather poor solvent for PMMA. It works much faster at 60 degrees, but don't try heating flammable solvents in your oven unless you are certain it is certified explosion-proof (most paraffin ovens are not). Even then, unless you have the oven in or at least next to a fume hood, you'll get a lot of xylene vapors in the air as a result of heating it. I deplasticise PMMA with 2-methoxyethyl acetate, which only takes a couple of hours at room temperature. This has a rather strong though not unpleasant odor, and should be used in the hood. _______________________________________________ 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 note 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: 6 Date: Tue, 13 Nov 2007 09:34:47 +0800 From: "San Tin" Subject: [Histonet] MyoD1 To: Message-ID: Content-Type: text/plain; charset="us-ascii" Dear All, Can anyone give me the protocol of MyoD1 clone 5.8A on Ventana IHC autostainer? I have problem with staining. Thanks San ------------------------------ Message: 7 Date: Tue, 13 Nov 2007 14:03:54 +1100 From: "Farish, Craig" Subject: [Histonet] RE: TGIF To: Message-ID: <79A000B60AFE8045BA2581119DFEC44402A251D4@ESWW01.CSUMain.csu.edu.au> Content-Type: text/plain; charset="us-ascii" And in return: A surgeon, pathologist, physician, psychiatrist and radiologist go duck hunting out on a pond. As the first duck flies over head, the psychiatrist takes aim, ....but doesn't fire. "What the hell happened?" demands the surgeon (they're known for their retiring manner) "Well, says the psychiatrist, I know its a duck, and YOU know its a duck - but does the duck know its a duck?" The second bird flies over head, and the physician takes aim.....but doesn't fire. "What now???" roars the surgeon. Well, says the physician, in a recent article I read in the New England Journal of Medicine, 12% of ducks are actually Terns, and they're a protected species.... The third duck flies over, and the radiologist takes aim......then paddles the boat further into the pond, then takes aim, then paddles back a bit, then takes a further aim....and by this time the duck has gone. "Jesus wept!" explodes the surgeon, what the f*** were you doing? Well, says the radiologist - every time I took aim, I realised that I needed another view. After lunch, they re-group for more hunting. This time the surgeon gets out his double barrelled, gold plated bazooka, blows his duck whistle - and as a flock of birds swarm over head, he fires randomly and enthusiastically into the air. Objects rain out of the sky. When he had finally finished blasting away to the heavens - he turned to the pathologist and said: "row over to all those bodies, and tell me if any of them was a duck" Craig (Joe) Farish Technical Officer School of Agricultural and Veterinary Sciences Charles Sturt University Wagga Wagga NSW Australia ' I don't want to achieve immortality through my work, I want to achieve immortality through not dying' - Woody Allen ------------------------------ Message: 8 Date: Tue, 13 Nov 2007 12:54:35 +0200 From: "K M" Subject: [Histonet] Claudin-1 as a prognostic factor in colorectal cancer To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hi all I am here again and I start do my Msc and the topic I have chosen is: Claudin-1 as a prognostic factor in colorectal cancer. I will tell you each step I will do so I can benefit from your valuable experiences and knowledge. Thanks again to all members of this valuable forum -- Khalaf Mobile Phone: 0020127920404 Egypt ------------------------------ Message: 9 Date: Tue, 13 Nov 2007 08:18:06 -0500 From: "MaryAnn Dixon" Subject: [Histonet] staining of negative controls with IHC To: Message-ID: <47395DBE.6EA2.00FD.0@vetmed.ufl.edu> Content-Type: text/plain; charset=US-ASCII We are currently retrieving animal tissue in a pressure cooker with a program of 125 degrees for 30 secs. This protocol was taken from biocare but I'm sure this is for human tissue. Most of the tissue is retrieved with Biocare's Reveal (pH 6.0). We are receiving some background staining of our negative controls whereas if we did not retrieve them, there is none. We were staining for KI-67 which is a nuclear stain. I am using a tonsil for the control. The negative control showed some sporatic cytoplasmic staining on the Universal negative control from biocare as well as deionized water. Since then we have taken the temperature down to 115 degrees and increased the time to 25 minutes. It seems to have decreased the cytoplasmic staining but there is still some lingering. Anyone out there have any ideas??? I'm sure there is a better protocol for retrieving animal tissues. It has to be a time and temperature thing! MaryAnn Dixon Biological Scientist Anatomic Pathology University of Florida School of Veterinary Medicine 352-392-2235 ext 4517 ------------------------------ Message: 10 Date: Tue, 13 Nov 2007 05:32:35 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] staining of negative controls with IHC To: MaryAnn Dixon , histonet@lists.utsouthwestern.edu Message-ID: <428387.48113.qm@web61222.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 There are many who prefer the pressure cooker because of the low incubation period but I personally did not like it because of background issues as you describe and because some tissues peel-off the slides (too much heat). Because of that I ended using a HIER step in a buffer (citrate or EDTA depending on the pH) initially heated to boliling temperature in a microwave oven and the actual HIER to take place in a steamer during 20 minutes + 20 minutes out of the steamer. All problems were resolved this way. Give it a try. Ren? J. MaryAnn Dixon wrote: We are currently retrieving animal tissue in a pressure cooker with a program of 125 degrees for 30 secs. This protocol was taken from biocare but I'm sure this is for human tissue. Most of the tissue is retrieved with Biocare's Reveal (pH 6.0). We are receiving some background staining of our negative controls whereas if we did not retrieve them, there is none. We were staining for KI-67 which is a nuclear stain. I am using a tonsil for the control. The negative control showed some sporatic cytoplasmic staining on the Universal negative control from biocare as well as deionized water. Since then we have taken the temperature down to 115 degrees and increased the time to 25 minutes. It seems to have decreased the cytoplasmic staining but there is still some lingering. Anyone out there have any ideas??? I'm sure there is a better protocol for retrieving animal tissues. It has to be a time and temperature thing! MaryAnn Dixon Biological Scientist Anatomic Pathology University of Florida School of Veterinary Medicine 352-392-2235 ext 4517 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Never miss a thing. Make Yahoo your homepage. ------------------------------ Message: 11 Date: Tue, 13 Nov 2007 10:39:46 EST From: AGrobe2555@aol.com Subject: Re: [Histonet] staining of negative controls with IHC To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" You don't say what detection system you are using....However, if you are using a Biotin-avidin/streptavidin detection, the background may be due to the exposure of endogenous biotin by HIER. You may need to incorporate an appropriate blocking agent/system Albert Albert C. Grobe, PhD International Heart Institute of Montana Foundation Tissue Engineering Lab, Saint Patrick Hospital ************************************** See what's new at http://www.aol.com ------------------------------ Message: 12 Date: Tue, 13 Nov 2007 08:24:00 -0800 From: "Tarango, Mark" Subject: RE: [Histonet] staining of negative controls with IHC To: "MaryAnn Dixon" , histonet@lists.utsouthwestern.edu Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6F9A@NVCIEXCH02.NVCI.org> Content-Type: text/plain; charset=us-ascii You probably don't even need to get up to boiling for ki-67. Try 30 minutes at 95 C in your pressure cooker. Even on a "negative" control you're bound to see something stain positive for ki-67, since it's a proliferation marker... although it shouldn't be cytoplasmic. I'm just guessing there's got to be a cell or two in the tissue that's dividing. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of MaryAnn Dixon Sent: Tuesday, November 13, 2007 5:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] staining of negative controls with IHC We are currently retrieving animal tissue in a pressure cooker with a program of 125 degrees for 30 secs. This protocol was taken from biocare but I'm sure this is for human tissue. Most of the tissue is retrieved with Biocare's Reveal (pH 6.0). We are receiving some background staining of our negative controls whereas if we did not retrieve them, there is none. We were staining for KI-67 which is a nuclear stain. I am using a tonsil for the control. The negative control showed some sporatic cytoplasmic staining on the Universal negative control from biocare as well as deionized water. Since then we have taken the temperature down to 115 degrees and increased the time to 25 minutes. It seems to have decreased the cytoplasmic staining but there is still some lingering. Anyone out there have any ideas??? I'm sure there is a better protocol for retrieving animal tissues. It has to be a time and temperature thing! MaryAnn Dixon Biological Scientist Anatomic Pathology University of Florida School of Veterinary Medicine 352-392-2235 ext 4517 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== ------------------------------ Message: 13 Date: Tue, 13 Nov 2007 11:01:02 -0500 From: "Lesley Bechtold" Subject: [Histonet] automated paraffin embedding systems To: "Histology Network" Message-ID: <20071113110102345.00000002152@spikey> Content-Type: text/plain; charset=us-ascii Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 ------------------------------ Message: 14 Date: Tue, 13 Nov 2007 09:03:48 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] automated paraffin embedding systems To: Lesley Bechtold , Histology Network Message-ID: <284972.24114.qm@web61211.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 This embedding instrument is manufactured by Sakura. It is not that the instrument orients the tissue, that is done when the tissues are placed in the cassette and held in place by a plastic frame that will be sectioned along with the tissue when the block is prepared. To function ideally it requires a special type of cassette and cassette holder (that is used alsao by the Sakura Xpress microwave tissue processor). Ren? J. Lesley Bechtold wrote: Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. ------------------------------ Message: 15 Date: Tue, 13 Nov 2007 11:09:57 -0600 From: "Mike Pence" Subject: RE: [Histonet] automated paraffin embedding systems To: "Rene J Buesa" , "Lesley Bechtold" , "Histology Network" Message-ID: <661949901A768E4F9CC16D8AF8F2838CA1C7A8@IS-E2K3.grhs.net> Content-Type: text/plain; charset="iso-8859-1" Is anyone out there using this function of the Sakura Xpress? Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, November 13, 2007 11:04 AM To: Lesley Bechtold; Histology Network Subject: Re: [Histonet] automated paraffin embedding systems This embedding instrument is manufactured by Sakura. It is not that the instrument orients the tissue, that is done when the tissues are placed in the cassette and held in place by a plastic frame that will be sectioned along with the tissue when the block is prepared. To function ideally it requires a special type of cassette and cassette holder (that is used alsao by the Sakura Xpress microwave tissue processor). Ren? J. Lesley Bechtold wrote: Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Tue, 13 Nov 2007 12:26:32 -0500 From: "Lesley Bechtold" Subject: RE: [Histonet] automated paraffin embedding systems To: "Mike Pence" , "Rene J Buesa" , "Histology Network" Message-ID: <20071113122632276.00000002152@spikey> Content-Type: text/plain; charset=iso-8859-1 Thank you to everyone for so quickly responding!!! I'm calling my Sakura rep for more info. Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 -----Original Message----- From: Mike Pence [mailto:mpence@grhs.net] Sent: Tuesday, November 13, 2007 12:10 PM To: Rene J Buesa; Lesley Bechtold; Histology Network Subject: RE: [Histonet] automated paraffin embedding systems Is anyone out there using this function of the Sakura Xpress? Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Tuesday, November 13, 2007 11:04 AM To: Lesley Bechtold; Histology Network Subject: Re: [Histonet] automated paraffin embedding systems This embedding instrument is manufactured by Sakura. It is not that the instrument orients the tissue, that is done when the tissues are placed in the cassette and held in place by a plastic frame that will be sectioned along with the tissue when the block is prepared. To function ideally it requires a special type of cassette and cassette holder (that is used alsao by the Sakura Xpress microwave tissue processor). Ren? J. Lesley Bechtold wrote: Hi, I've heard rumors that there are systems for doing automated paraffin embedding AND that they orient tissue correctly. Could someone (vendor, customer, anyone) please point me in the right direction? Thank you! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Tue, 13 Nov 2007 12:32:49 -0500 From: "Christine I. Braaten" Subject: [Histonet] Antibody cocktails To: Message-ID: Content-Type: text/plain; charset="us-ascii" Can anybody tell me if there is another company besides Biocare that offers an antibody cocktail with P504S, HMW CK, and p63? Does anybody use the Biocare antibody detection system? I'd appreciate some feedback. Thanks. CONFIDENTIALITY NOTICE: This electronic message, including any attachments, is for the sole use of the intended recipients 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 electronic mail and destroy all copies of the original message. ------------------------------ Message: 18 Date: Tue, 13 Nov 2007 12:40:56 -0500 (EST) From: "FU,DONGTAO" Subject: RE: [Histonet] staining of negative controls with IHC To: Histonet@lists.utsouthwestern.edu Message-ID: <1798590190.34131194975656578.JavaMail.osg@osgjas03.cns.ufl.edu> Content-Type: text/plain; format=flowed; charset=us-ascii Hi, Ki67 staining in both human and mouse tissue in our lab works very well. We use steamer method 30min in Citra buffer. Ann Dongtao Fu MD, Ph.D Lab Manager Molecular Pathology Core Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 On Tue Nov 13 11:24:00 EST 2007, "Tarango, Mark" wrote: > You probably don't even need to get up to boiling for ki-67. Try > 30 > minutes at 95 C in your pressure cooker. Even on a "negative" > control > you're bound to see something stain positive for ki-67, since > it's a > proliferation marker... although it shouldn't be cytoplasmic. > I'm just > guessing there's got to be a cell or two in the tissue that's > dividing. > > > Mark Adam Tarango HT(ASCP) > Histology & IHC Supervisor > Nevada Cancer Institute > One Breakthrough Way > Las Vegas, NV 89135 > Direct Line (702) 822-5112 > Treo (702) 759-9229 > Fax (702) 939-7663 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > MaryAnn > Dixon > Sent: Tuesday, November 13, 2007 5:18 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] staining of negative controls with IHC > > We are currently retrieving animal tissue in a pressure cooker > with a > program of 125 degrees for 30 secs. This protocol was taken from > biocare but I'm sure this is for human tissue. Most of the tissue > is > retrieved with Biocare's Reveal (pH 6.0). We are receiving some > background staining of our negative controls whereas if we did > not > retrieve them, there is none. We were staining for KI-67 which > is a > nuclear stain. I am using a tonsil for the control. The negative > control > showed some sporatic cytoplasmic staining on the Universal > negative > control from biocare as well as deionized water. Since then we > have > taken the temperature down to 115 degrees and increased the time > to 25 > minutes. It seems to have decreased the cytoplasmic staining but > there > is still some lingering. Anyone out there have any ideas??? I'm > sure > there is a better protocol for retrieving animal tissues. It has > to be a > time and temperature thing! > MaryAnn Dixon > Biological Scientist > Anatomic Pathology > University of Florida > School of Veterinary Medicine > 352-392-2235 ext 4517 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > "EMF " made the following annotations. > ------------------------------------------------------------------------------ > CONFIDENTIALITY NOTICE: This e-mail message, including any > attachments, is for the sole use of the intended recipient(s) and > may contain confidential, proprietary, and/or privileged > information protected by law. If you are not the intended > recipient, you may not use, copy, or distribute this e-mail > message or its attachments. If you believe you have received this > e-mail message in error, please contact the sender by reply > e-mail and destroy all copies of the original message > ============================================================================== > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 17 **************************************** From jmahoney <@t> alegent.org Mon Nov 19 11:27:55 2007 From: jmahoney <@t> alegent.org (Janice Mahoney) Date: Mon Nov 19 11:28:17 2007 Subject: [Histonet] FS Stain kit Message-ID: <4741733B0200003C000212CE@gwia.alegent.org> HELP! Does anyone know of an H&E Frozen Section staining kit (other than the Shandon one that is on back order)? I have quite a few Hospital sites where there are no HT's or anyone interested enough to change stains and reagents for FS. Therefore we rely on the rapid H&E kits. Thanks for the help. Jan Omaha, NE From rjr6 <@t> psu.edu Mon Nov 19 11:47:15 2007 From: rjr6 <@t> psu.edu (Roberta Horner) Date: Mon Nov 19 11:47:23 2007 Subject: [Histonet] Histology Strike! In-Reply-To: <875640.56798.qm@web52511.mail.re2.yahoo.com> References: <875640.56798.qm@web52511.mail.re2.yahoo.com> Message-ID: I plan on taking it even further. I am starting on Wednesday and I have no back up. I have been begging for help for years. Roberta Horner HT/HTL Animal Diagnostic Lab Penn State University From Cutter2754 <@t> aol.com Mon Nov 19 12:12:03 2007 From: Cutter2754 <@t> aol.com (Cutter2754@aol.com) Date: Mon Nov 19 12:12:14 2007 Subject: [Histonet] atlanta jobs Message-ID: Hi all, I'm moving to the Atlanta area in a few months and would like to know about the employment situation. I found several jobs available but would also like to hear about people's experiences - the good and the bad! Thanks, Kathy ************************************** See what's new at http://www.aol.com From doug <@t> ppspath.com Mon Nov 19 12:13:33 2007 From: doug <@t> ppspath.com (Douglas D Deltour) Date: Mon Nov 19 12:14:21 2007 Subject: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer In-Reply-To: <8854ff80711190922y18de3ca0u1219e842605867e6@mail.gmail.com> Message-ID: Invitrogen-Zymed has a pre-dilute that can be used in a prep-kit on the XT. This is the same one that I am currently troubleshooting. Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sate Hamza Sent: Monday, November 19, 2007 12:23 PM To: Douglas D Deltour; Histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer Thanks to all who have responded to my message. I asked my colleague and she said that she has not ordered any antibody yet. She said she wanted to see if she can get a protocol first before she orders the antibody. She was wondering what the source of your antibody was .. Thanks again .. Sate On 11/19/07, Douglas D Deltour wrote: > > We just started using the MITF-1 antibody on the XT. I got it validated > and > we were running it for about 2 months and then all of a sudden it stopped > working, but that is another story. Which company did your colleague get > the > antibody from? I used the same protocol that I used for the MART-1 for > validation. This was working great for the MITF until..... > > [---] > > > > -- Sate Hamza, MD, FRCPC Dermatopathologist Winnipeg, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From m5johnso <@t> ucsd.edu Mon Nov 19 12:14:31 2007 From: m5johnso <@t> ucsd.edu (Johnson, Mindy) Date: Mon Nov 19 12:14:36 2007 Subject: [Histonet] Metallic sheen on hematoxylin Message-ID: Ok, I was trying to find where I found this information, but I can't find it, so.... The metallic sheen I get on top of my hematoxylin, I though was very concentrated (this is the information I was talking about). Is this true and if so, what can I mix it with to extend the life of it? Thanks! Mindy Medical Teaching Labs, UCSD From ATOJSU <@t> aol.com Mon Nov 19 13:07:28 2007 From: ATOJSU <@t> aol.com (ATOJSU@aol.com) Date: Mon Nov 19 13:07:35 2007 Subject: [Histonet] re:B-5 solution Message-ID: We have 1 Pathologist who insists on using B-5 solution for nuclear detail on small biopsies. How many labs are still using B 5 or what is you suggestion other than Zinc Formalin for fixation. Thanks. ************************************** See what's new at http://www.aol.com From b-frederick <@t> northwestern.edu Mon Nov 19 13:20:53 2007 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Mon Nov 19 13:21:12 2007 Subject: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer In-Reply-To: <20071119181735.7FCF268FF1@slipkid.it.northwestern.edu> Message-ID: <000001c82ae1$50b449e0$d00f7ca5@lurie.northwestern.edu> LabVision also has a pre-dilute for clone D5,which is what we use undiluted. Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Douglas D Deltour Sent: Monday, November 19, 2007 12:14 PM To: 'Sate Hamza'; Histonet@lists.utsouthwestern.edu Subject: RE: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer Invitrogen-Zymed has a pre-dilute that can be used in a prep-kit on the XT. This is the same one that I am currently troubleshooting. Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sate Hamza Sent: Monday, November 19, 2007 12:23 PM To: Douglas D Deltour; Histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer Thanks to all who have responded to my message. I asked my colleague and she said that she has not ordered any antibody yet. She said she wanted to see if she can get a protocol first before she orders the antibody. She was wondering what the source of your antibody was .. Thanks again .. Sate On 11/19/07, Douglas D Deltour wrote: > > We just started using the MITF-1 antibody on the XT. I got it validated > and > we were running it for about 2 months and then all of a sudden it stopped > working, but that is another story. Which company did your colleague get > the > antibody from? I used the same protocol that I used for the MART-1 for > validation. This was working great for the MITF until..... > > [---] > > > > -- Sate Hamza, MD, FRCPC Dermatopathologist Winnipeg, Canada _______________________________________________ Histonet mailing list Histonet@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 rjbuesa <@t> yahoo.com Mon Nov 19 14:24:36 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 19 14:24:41 2007 Subject: [Histonet] Metallic sheen on hematoxylin In-Reply-To: Message-ID: <414329.50634.qm@web61217.mail.yahoo.com> Metallic sheen on the surface of the hematoxylin is due to a surface oxydation. The best way of dealing with it is to take a kin-wip and skim the surface with it. Another solution (more cumbersome) is to filter the hematoxylin. Do not mix anything with the hematoxylin because then you will end with a different formula. This is a surface chemical product that can be eliminated mechanically. Ren? J. "Johnson, Mindy" wrote: Ok, I was trying to find where I found this information, but I can't find it, so.... The metallic sheen I get on top of my hematoxylin, I though was very concentrated (this is the information I was talking about). Is this true and if so, what can I mix it with to extend the life of it? Thanks! Mindy Medical Teaching Labs, UCSD _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. From chiggerson <@t> memhosp.com Mon Nov 19 15:02:25 2007 From: chiggerson <@t> memhosp.com (chiggerson@memhosp.com) Date: Mon Nov 19 15:02:37 2007 Subject: [Histonet] Cindy Higgerson is out of the office. Message-ID: I will be out of the office starting Mon 11/19/2007 and will not return until Tue 11/27/2007. I will respond to your message when I return. From CIngles <@t> uwhealth.org Mon Nov 19 15:15:02 2007 From: CIngles <@t> uwhealth.org (Ingles Claire) Date: Mon Nov 19 15:16:18 2007 Subject: SPAM-LOW: Re: [Histonet] no subject References: <20071119150200.D376C300C4@mailsec2-in.hosp.wisc.edu> Message-ID: <08A0A863637F1349BBFD83A96B27A50A1200A2@uwhis-xchng3.uwhis.hosp.wisc.edu> I am! I'll even boycott Friday too! Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Douglas D Deltour Sent: Mon 11/19/2007 9:01 AM To: 'Emily Sours'; histonet@lists.utsouthwestern.edu Subject: RE: SPAM-LOW: Re: [Histonet] no subject I honor of this thread I declare Thursday November 22nd National Histology Strike Day! We will not report to work on this day to show our dissatisfaction. Who is with me? From AGrobe2555 <@t> aol.com Mon Nov 19 17:04:33 2007 From: AGrobe2555 <@t> aol.com (AGrobe2555@aol.com) Date: Mon Nov 19 17:04:42 2007 Subject: [Histonet] IHC for MMP2, MMP9, TIMP1 and TIMP2? Message-ID: Hello all, I have a panel of tissues (Large and small intestine, liver, spleen, kidney, heart, esophagus, lung) as well as normal artery and vein tissue (all are from sheep) that we are using to test/optimize IHC for MMP2, MMP9, TIMP1 and TIMP2. We have stained for MMP9 so far, and see some staining in cells (appear to be macrophages) located in the small and large intestine, and in the adventitial/ablumenal tissue of the vein. We will be doing the other stains in the days to come. My difficulty is that I am in unknown territory with these antibodies. Has anyone of you stained for these before? What should I be looking for as a positive signal in these tissues? Can anyone point me to some reference images? In the interim, I will continue to look for information as the product sheets are not really providing information as to appropriate + control tissues. Thank you, Albert Albert C. Grobe, PhD International Heart Institute of Montana Foundation Tissue Engineering Lab, Saint Patrick Hospital ************************************** See what's new at http://www.aol.com From lpwenk <@t> sbcglobal.net Mon Nov 19 18:33:31 2007 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Mon Nov 19 18:33:43 2007 Subject: [Histonet] re:B-5 solution In-Reply-To: Message-ID: <001201c82b0c$faad1c30$0202a8c0@HPPav2> Why not zinc formalin? Usually, if people have tried zinc formalin and didn't like it, it's because they didn't leave the tissue in the zinc formalin long enough. Usually they have left it for the same amount of time as the B5, and as a result the nuclear detail is lousy. The secret is that zinc salts binds a little slower than mercuric chloride. So take your B5 time, and multiply it by 1.5, and that will be a good starting point for zinc formalin. In other words, if the biopsies were fixed in B5 in 2 hours, then 2 x 1.5 = 3 hours would be a good guess at how long to fix in zinc formalin. Peggy A. 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 ATOJSU@aol.com Sent: Monday, November 19, 2007 2:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re:B-5 solution We have 1 Pathologist who insists on using B-5 solution for nuclear detail on small biopsies. How many labs are still using B 5 or what is you suggestion other than Zinc Formalin for fixation. Thanks. ************************************** See what's new at http://www.aol.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peddinti_2002us <@t> yahoo.co.in Mon Nov 19 19:28:23 2007 From: peddinti_2002us <@t> yahoo.co.in (kamal prasad) Date: Mon Nov 19 19:28:32 2007 Subject: [Histonet] (no subject) Message-ID: <728317.58538.qm@web8705.mail.in.yahoo.com> Yes , 10% formic acid is best decalcifier it will keep cell morphology better. Formic acod need to mix it with 10%NBF . --------------------------------- Forgot the famous last words? Access your message archive online. Click here. From tgoodpas <@t> fhcrc.org Mon Nov 19 19:37:40 2007 From: tgoodpas <@t> fhcrc.org (Goodpaster, Tracy A) Date: Mon Nov 19 19:37:46 2007 Subject: [Histonet] QIHC In-Reply-To: <5AEC610C1CE02945BD63A395BA763EDE011B6F92@NVCIEXCH02.NVCI.org> References: <5AEC610C1CE02945BD63A395BA763EDE011B6F92@NVCIEXCH02.NVCI.org> Message-ID: I am also interested in this information. I don't know how specific it will be, especially over clinical lab regulations. Any additional information would be helpful! Thanks! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tarango, Mark Sent: Sunday, November 11, 2007 3:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] QIHC To anyone who has taken the QIHC exam, what kinds of questions does it ask? I've got a few books from the reading list, but I have no idea where to focus my attention. It is more about staining techniques or immunophenotyping? Quality control stuff? What's on the test that you might not expect? Is it a difficult exam? Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 "EMF " made the following annotations. ------------------------------------------------------------------------ ------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ======================================================================== ====== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ombadda3 <@t> gmail.com Tue Nov 20 03:28:27 2007 From: ombadda3 <@t> gmail.com (K M) Date: Tue Nov 20 03:28:36 2007 Subject: [Histonet] Prognostic factors in colorectal cancers Message-ID: Dear colleagues I am doing study on IHC in colorectal cancers in human tissues and correlate them with prognosis .I used an antibody for Claudin-1(a tight junction protein found in cell membrane) as a prognostic factor.Ineed another prognostic factor for colorectal cancer for that I am enquiring if any one know other strong prognostic factor used in colorectal cancers? Dr.K.M.Omer From jnocito <@t> satx.rr.com Tue Nov 20 06:08:56 2007 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Tue Nov 20 06:08:58 2007 Subject: [Histonet] forceps and bunsen burners References: <037a01c829ff$0cc44dd0$6d01a8c0@CHERYLSLAPTOP> Message-ID: <015d01c82b6e$207b2f40$0302a8c0@yourxhtr8hvc4p> the good ole days. The lab I'm at now used to used micro loop flamers. It was like playing the game "operation", if you touched the sides, sparks shoot up. It does cause a nice light show with the lights off. My other place of employment, we used alcohol burners, others used gauze. Personally, I miss the days of having a cigarette hanging from my mouth while cutting and coverslipping. I admit, I'm much healthier today, but miss the good ole days. JTT ----- Original Message ----- From: "Cheryl R. Kerry" To: Sent: Sunday, November 18, 2007 10:21 AM Subject: [Histonet] forceps and bunsen burners >I really miss Bunsen burners! Now we use a lot of kimwipes. > > Kemlo--I recall the days of wiping your embedding center with a xylene > soaked rag...me with a cuppa joe on my 'tome and others with cigarettes > hanging out the sides of their mouths...and yes, the occasional lab fires > that resulted!! And I really miss REAL light green SF yellowish! > > Cheryl > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kemlo > Sent: Sunday, November 18, 2007 4:22 AM > To: 'sheila adey'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] (no subject) > > In the good old days of Bunsen burners we used to fry the end of the > forceps; nothing survived that and if it did it was readily identifiable. > Alas the Bunsen burner has been consigned to the politically incorrect as > the 'Scientists' of today would incinerate themselves whilst the > 'Technicians' of yesterday didn't (well not often). > > The sad demise of mercury, lead, Bunsen burners, formalin, anything too > hot, > too cold, too explosive, too poisonous, etc. Having your tea in the Lab > next > to the specimens and processing TB specimens 'on the bench'. > > Would Histology have the techniques and stains it now has if harmful > chemicals had not been experimented with? Will anything new be discovered > by > the HistoTech if all that we can use is 'safe' chemicals and procedures? I > don't see the kids fiddling with things like we use to, no explosions, no > fires and no injuries. Am I just an old reactionary waiting to be put out > to > pasture and ruminate on what was? > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of sheila > adey > Sent: 17 November 2007 20:41 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] (no subject) > > > Hi Netters, > > We are trying to minimize possible embedding contaminations. What are > other > people doing to prevent contamination due to forceps etc. > > Thanks in advanceSheila Adey HT MLTPort Huron HospitalMichigan > _________________________________________________________________ > Are you ready for Windows Live Messenger Beta 8.5 ? Get the latest for > free > today! > http://entertainment.sympatico.msn.ca/WindowsLiveMessenger__________________ > _____________________________ > Histonet mailing list > Histonet@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 victor <@t> pathology.washington.edu Tue Nov 20 10:08:36 2007 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Tue Nov 20 10:08:44 2007 Subject: [Histonet] New RA Lamb Cassette Printer UK Only Message-ID: <47430684.4000608@pathology.washington.edu> Has anyone purchased or seen this new cassette labeler? We are still waiting for Thermo to release the US version. I have heard it will print cassettes with bar codes in 2.5 seconds, which is remarkable. Victor -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. From tara.ollis <@t> teamstaffrx.com Tue Nov 20 10:48:11 2007 From: tara.ollis <@t> teamstaffrx.com (Tara Ollis) Date: Tue Nov 20 10:48:14 2007 Subject: [Histonet] Histologist Job in ATL GA Message-ID: <18512542.1195577290900.JavaMail.cfservice@webserver29> Hello- I have a Histologist position available in a dermatologist office in the greater Atlanta area. They need someone with MOHS experience and prefer HTL Certification. Feel free to email me your resume or call me for more details. Thanks! Tara Ollis | TeamStaff Rx, Inc. Office: 1.877.523.9897 ext.5511 | www.teamstaffrx.com From MVaughan4 <@t> ucok.edu Tue Nov 20 11:54:45 2007 From: MVaughan4 <@t> ucok.edu (MVaughan4@ucok.edu) Date: Tue Nov 20 11:55:14 2007 Subject: [Histonet] Matrigel as culture medium - try agarose embedding In-Reply-To: Message-ID: Teri, Can you embed or surround the matrigel mixture in a low-melting agarose first, then postfix the agarose/matrigel mixture in formalin, followed by paraffin embedding? I have used it with collagen matrices. Mel Melville B. Vaughan, Ph. D. Assistant Professor of Biology Campus Coordinator, NSF Sure-Step program University of Central Oklahoma 100 N. University Drive Edmond, OK 73034 http://www.biology.ucok.edu/PersonalPages/mvaughan/Default.htm Message: 8 Date: Fri, 16 Nov 2007 14:42:54 -0600 From: "Keller, Charles" Subject: RE: [Histonet] Matrigel as culture medium To: "Johnson, Teri" , Message-ID: Content-Type: text/plain; charset="us-ascii" Dear Teri, a possible solution is to use Extracel (from Glycosan Inc in Utah) instead of Matrigel. I've fixed Extracel in 10% formalin and sectioned in in paraffin. It has the advantage of being synthetic (Matrigel has viral contamination from time to time). Sincerely, Charles -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Friday, November 16, 2007 9:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Matrigel as culture medium One of our researchers is using matrigel as a culture medium on chambered slides. We tried paraffin embedding them, and found the Matrigel wasn't really solid enough to hold the cellular structure in place. We found it particularly difficult to keep some sort of consistency scraping it off (it had the consistency of warm jelly). I'd love to hear some ideas on how we can accomplish sample processing without centrifugation. Ideally it would be good to encase the cells in a matrix that would solidfy enough to embed as a cell block, keeping the cellular structure intact. Thanks for your help! Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------- **CONFIDENTIALITY** -This email (including any attachments) may contain confidential, proprietary and privileged information. Any unauthorized disclosure or use of this information is prohibited. From jm.lapointe <@t> accellab.com Tue Nov 20 12:17:04 2007 From: jm.lapointe <@t> accellab.com (Jean-Martin Lapointe) Date: Tue Nov 20 12:21:06 2007 Subject: [Histonet] IHC for MMP2, MMP9, TIMP1 and TIMP2? Message-ID: Probably not the info you want, but we have tried here to set up IHC for MMP9 in rabbit paraffin-embedded tissue, using a few different commercial antibodies, but we abandoned it because I didn't feel the staining we had was specific. __________________________________ Jean-Martin Lapointe, DMV, MS, dACVP Vice-President, Pathologie AccelLAB Inc 1635 Lionel-Bertrand, Boisbriand Qu?bec, Canada J7H 1N8 tel: 450-435-9482 ext.247 fax: 450-435-4795 jm.lapointe@accellab.com From DixonM <@t> vetmed.ufl.edu Tue Nov 20 13:16:08 2007 From: DixonM <@t> vetmed.ufl.edu (MaryAnn Dixon) Date: Tue Nov 20 13:16:38 2007 Subject: [Histonet] IgA, IgM, IgG Message-ID: <4742EC29.6EA2.00FD.0@vetmed.ufl.edu> I am working up anti dog IgA, IgG and IgM antibodies and was wondering what the better control tissues would be for these. I have been using tonsil for all three but would love some input on any that work better. Also, what is everyone using for pretreatment? I'm teetering between prot. K and no treatment at all. The dilutions of these antibodies are getting higher too. My IgG was out to 1:6000. Do these antibodies generally have some accepted background with them? I thought I was told that they do and you cant get it out. Can anyone please share some dilutions and pretreatments for these 3 antibodies. Don't care if they are for anti-human, just would like a starting point. I am using HRP mach 4 detection from biocare. MaryAnn Dixon Biological Scientist Anatomic Pathology University of Florida School of Veterinary Medicine 352-392-2235 ext 4517 From CIngles <@t> uwhealth.org Tue Nov 20 13:23:11 2007 From: CIngles <@t> uwhealth.org (Ingles Claire) Date: Tue Nov 20 13:23:21 2007 Subject: [Histonet] no subject References: <119713.97956.qm@web50111.mail.re2.yahoo.com> Message-ID: <08A0A863637F1349BBFD83A96B27A50A1200A4@uwhis-xchng3.uwhis.hosp.wisc.edu> Sadly I am already in a union. Granted it is with the custodians and food service people. I have no idea WHY, by the way... Our lab manager is also a nurse, so she knows &%&% about lab stuff. Fortunately she knows she doesn't know and trusts us to know what we are doing, and why we are doing it. So we're pretty much left to our own devices. (HAHAHA THE FOOLS!...) Keep fighting the power though. Every little breakthrough helps. Claire Ingles UW Hospital Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Emily Sours Sent: Mon 11/19/2007 8:45 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] no subject Union! Union! Union! From am3309 <@t> uga.edu Tue Nov 20 13:25:17 2007 From: am3309 <@t> uga.edu (Abigail M. Butler) Date: Tue Nov 20 13:25:31 2007 Subject: [Histonet] RE: QIHC Message-ID: <20071120142517.JIJ06459@punts1.cc.uga.edu> I was just responding to the questions about the QIHC exam. It's not as difficult as you may anticipate. And I really don't think studying really helped me much. I think you just have to know the stuff that you do. Now, if you are not currently performing IHC you probably do need to study. But if IHC is something that you do on a daily basis then you should be fine. Also, to the guy that has the title IHC supervisor, how did you get that title? And how many techs do you have under you? If you don't mind me asking...... Abbie Butler, HT (ASCP), QIHC University of Georgia College of Veterinary Medicine From lhotaks <@t> mcmaster.ca Tue Nov 20 13:25:36 2007 From: lhotaks <@t> mcmaster.ca (Sarka Lhotak) Date: Tue Nov 20 13:26:04 2007 Subject: [Histonet] IHC for MMP2, MMP9, TIMP1 and TIMP2 Message-ID: Hi Albert, I did IHC for MMP2, MMP9, TIMP1, TIMP2, and also MT1-MMP, on human tissues from breast carcinoma and bone metastasis from breast carcinoma (i.e. in human tissue). It was published in Clinical and Experimental Metastasis, 2000, 18(6):463-70. I believe the staining protocols and antibodies used were described in sufficient detail there. I would expect macrophages to stain for MMP-9, that's what I've seen as well (and osteoclasts). Of course, I am not sure to what extent will these antibodies work in sheep tissue. Good luck, check that paper, if you have questions, let me know. I would have to dig deep into my log books, it's been a while. Regards, Sarka Lhotak, PhD McMaster University, Hamilton, Ontario lhotaks@mcmaster.ca From shive003 <@t> umn.edu Tue Nov 20 13:42:32 2007 From: shive003 <@t> umn.edu (Jan Shivers) Date: Tue Nov 20 13:42:36 2007 Subject: [Histonet] IgA, IgM, IgG References: <4742EC29.6EA2.00FD.0@vetmed.ufl.edu> Message-ID: <007401c82bad$7e6a2040$b0065486@auxs.umn.edu> MaryAnn, I use tonsil control for dog IgG and IgM; I use stomach control (mucosa) for dog IgA. Your dilutions of Abs will, of course, depend upon what the original concentration is of your stock Ab and vendor prep. However, not all companies list the stock immunoglobulin concentration of their antibodies, so you'll just have to work them up and optimize dilutions for best staining results. Here's what I use currently, but these change with change of vendor and change of Ab lot number: Dog IgG - 1:6000; 0.4ug/ml working concentration Dog IgM - 1:4000; (no stock Ig concentration given by manuf.) Dog IgG - 1:1200; (no stock Ig concentration given by manuf.) I use Proteinase K (Dako) enzyme digestion for 5', room temp, to unmask antigens for all 3 antibodies. Yes, you'll get background with these specific antibodies, but you can decrease it by adding 4% normal dog serum in your 'linking or secondary' Ab (per volume). Hope this helps. Jan Shivers Section Head - IHC/Histo/EM Veterinary Diagnostic Laboratory College of Veterinary Medicine University of Minnesota St. Paul, MN shive003@umn.edu ----- Original Message ----- From: "MaryAnn Dixon" To: Sent: Tuesday, November 20, 2007 1:16 PM Subject: [Histonet] IgA, IgM, IgG I am working up anti dog IgA, IgG and IgM antibodies and was wondering what the better control tissues would be for these. I have been using tonsil for all three but would love some input on any that work better. Also, what is everyone using for pretreatment? I'm teetering between prot. K and no treatment at all. The dilutions of these antibodies are getting higher too. My IgG was out to 1:6000. Do these antibodies generally have some accepted background with them? I thought I was told that they do and you cant get it out. Can anyone please share some dilutions and pretreatments for these 3 antibodies. Don't care if they are for anti-human, just would like a starting point. I am using HRP mach 4 detection from biocare. MaryAnn Dixon Biological Scientist Anatomic Pathology University of Florida School of Veterinary Medicine 352-392-2235 ext 4517 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emerald_lake77 <@t> yahoo.com Tue Nov 20 15:21:38 2007 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Tue Nov 20 15:21:53 2007 Subject: [Histonet] Endothelial Cell Marker in FFPE Mouse Tissue Message-ID: <281567.9020.qm@web31706.mail.mud.yahoo.com> Hello, Holiday wishes to all of you. What are people using for endothelial cells in FFPE mouse tissue nowadays? Currently I am using P-selectin but want to find a better marker or two - preferrably CD31. Any suggestions would be very helpful. Thanks. Gustave Gustave T. Hebert Scientist II Wyeth - Research Cambridge MA --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From pierre.chaumat <@t> alphelys.com Tue Nov 20 15:38:58 2007 From: pierre.chaumat <@t> alphelys.com (Pierre CHAUMAT) Date: Tue Nov 20 15:39:15 2007 Subject: [Histonet] Formaline-free fixative In-Reply-To: <0MKqpg-1IuXbI10hY-0008EY@mx.kundenserver.de> Message-ID: <0ML2xA-1Iuao51g7u-0005Ac@mrelayeu.kundenserver.de> Hello all, We have developed a new fixative named RCL2 that provides a molecular quality close to frozen sections but with the morphology obtained with formaline. IHC requires much less retrieval, naturally. We have demonstrated stability of samples over 7 years. RCL2 also replaces advantageously deep freezing as RCL2 fixed tissues can be stored simply at -20?C thus generating large costs savings and simplicity for biobanking. Penetration speed of RCL2 is similar to formaline but you can leave your tissues into RCL2 without any damages on morphology nor signals. And to keep it for the end...RCL2 is absolutely non toxic and even better : drinkable for a "strong" man. The content ? A patented complex carbohydrate, acetic acid at 6% and ethanol at 65% (thus the "strong" man). We have hospitals and private labs starting to use it in routine. I want also to add that the French Cancer Research Institute is starting to test it for Her2neu after having given up on many other proposed fixatives with glyoxal based fixatives amongst them because of over fixation. Anyone here interested in testing it ? Motivated to change to a new fixative for real benefits ? Kind regards Pierre -----Message d'origine----- De : histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] De la part de histonet-request@lists.utsouthwestern.edu Envoy? : mardi 20 novembre 2007 19:14 ? : histonet@lists.utsouthwestern.edu Objet : Histonet Digest, Vol 48, Issue 28 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. atlanta jobs (Cutter2754@aol.com) 2. RE: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer (Douglas D Deltour) 3. Metallic sheen on hematoxylin (Johnson, Mindy) 4. re:B-5 solution (ATOJSU@aol.com) 5. RE: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer (Bernice Frederick) 6. Re: Metallic sheen on hematoxylin (Rene J Buesa) 7. Cindy Higgerson is out of the office. (chiggerson@memhosp.com) 8. RE: SPAM-LOW: Re: [Histonet] no subject (Ingles Claire) 9. IHC for MMP2, MMP9, TIMP1 and TIMP2? (AGrobe2555@aol.com) 10. RE: re:B-5 solution (Lee & Peggy Wenk) 11. (no subject) (kamal prasad) 12. RE: QIHC (Goodpaster, Tracy A) 13. Prognostic factors in colorectal cancers (K M) 14. Re: forceps and bunsen burners (Joe Nocito) 15. New RA Lamb Cassette Printer UK Only (Victor Tobias) 16. Histologist Job in ATL GA (Tara Ollis) 17. Matrigel as culture medium - try agarose embedding (MVaughan4@ucok.edu) ---------------------------------------------------------------------- Message: 1 Date: Mon, 19 Nov 2007 13:12:03 EST From: Cutter2754@aol.com Subject: [Histonet] atlanta jobs To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Hi all, I'm moving to the Atlanta area in a few months and would like to know about the employment situation. I found several jobs available but would also like to hear about people's experiences - the good and the bad! Thanks, Kathy ************************************** See what's new at http://www.aol.com ------------------------------ Message: 2 Date: Mon, 19 Nov 2007 13:13:33 -0500 From: "Douglas D Deltour" Subject: RE: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer To: "'Sate Hamza'" , Message-ID: Content-Type: text/plain; charset="us-ascii" Invitrogen-Zymed has a pre-dilute that can be used in a prep-kit on the XT. This is the same one that I am currently troubleshooting. Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sate Hamza Sent: Monday, November 19, 2007 12:23 PM To: Douglas D Deltour; Histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer Thanks to all who have responded to my message. I asked my colleague and she said that she has not ordered any antibody yet. She said she wanted to see if she can get a protocol first before she orders the antibody. She was wondering what the source of your antibody was .. Thanks again .. Sate On 11/19/07, Douglas D Deltour wrote: > > We just started using the MITF-1 antibody on the XT. I got it validated > and > we were running it for about 2 months and then all of a sudden it stopped > working, but that is another story. Which company did your colleague get > the > antibody from? I used the same protocol that I used for the MART-1 for > validation. This was working great for the MITF until..... > > [---] > > > > -- Sate Hamza, MD, FRCPC Dermatopathologist Winnipeg, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Mon, 19 Nov 2007 10:14:31 -0800 From: "Johnson, Mindy" Subject: [Histonet] Metallic sheen on hematoxylin To: "histonet@lists.utsouthwestern.edu" Message-ID: Content-Type: text/plain; charset="us-ascii" Ok, I was trying to find where I found this information, but I can't find it, so.... The metallic sheen I get on top of my hematoxylin, I though was very concentrated (this is the information I was talking about). Is this true and if so, what can I mix it with to extend the life of it? Thanks! Mindy Medical Teaching Labs, UCSD ------------------------------ Message: 4 Date: Mon, 19 Nov 2007 14:07:28 EST From: ATOJSU@aol.com Subject: [Histonet] re:B-5 solution To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" We have 1 Pathologist who insists on using B-5 solution for nuclear detail on small biopsies. How many labs are still using B 5 or what is you suggestion other than Zinc Formalin for fixation. Thanks. ************************************** See what's new at http://www.aol.com ------------------------------ Message: 5 Date: Mon, 19 Nov 2007 13:20:53 -0600 From: "Bernice Frederick" Subject: RE: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer To: "'Douglas D Deltour'" , "'Sate Hamza'" , Message-ID: <000001c82ae1$50b449e0$d00f7ca5@lurie.northwestern.edu> Content-Type: text/plain; charset="us-ascii" LabVision also has a pre-dilute for clone D5,which is what we use undiluted. Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Douglas D Deltour Sent: Monday, November 19, 2007 12:14 PM To: 'Sate Hamza'; Histonet@lists.utsouthwestern.edu Subject: RE: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer Invitrogen-Zymed has a pre-dilute that can be used in a prep-kit on the XT. This is the same one that I am currently troubleshooting. Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sate Hamza Sent: Monday, November 19, 2007 12:23 PM To: Douglas D Deltour; Histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: Re: [Histonet] MiTF on a Ventana Immunostainer Thanks to all who have responded to my message. I asked my colleague and she said that she has not ordered any antibody yet. She said she wanted to see if she can get a protocol first before she orders the antibody. She was wondering what the source of your antibody was .. Thanks again .. Sate On 11/19/07, Douglas D Deltour wrote: > > We just started using the MITF-1 antibody on the XT. I got it validated > and > we were running it for about 2 months and then all of a sudden it stopped > working, but that is another story. Which company did your colleague get > the > antibody from? I used the same protocol that I used for the MART-1 for > validation. This was working great for the MITF until..... > > [---] > > > > -- Sate Hamza, MD, FRCPC Dermatopathologist Winnipeg, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Mon, 19 Nov 2007 12:24:36 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Metallic sheen on hematoxylin To: "Johnson, Mindy" , "histonet@lists.utsouthwestern.edu" Message-ID: <414329.50634.qm@web61217.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Metallic sheen on the surface of the hematoxylin is due to a surface oxydation. The best way of dealing with it is to take a kin-wip and skim the surface with it. Another solution (more cumbersome) is to filter the hematoxylin. Do not mix anything with the hematoxylin because then you will end with a different formula. This is a surface chemical product that can be eliminated mechanically. Reni J. "Johnson, Mindy" wrote: Ok, I was trying to find where I found this information, but I can't find it, so.... The metallic sheen I get on top of my hematoxylin, I though was very concentrated (this is the information I was talking about). Is this true and if so, what can I mix it with to extend the life of it? Thanks! Mindy Medical Teaching Labs, UCSD _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. ------------------------------ Message: 7 Date: Mon, 19 Nov 2007 15:02:25 -0600 From: chiggerson@memhosp.com Subject: [Histonet] Cindy Higgerson is out of the office. To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII I will be out of the office starting Mon 11/19/2007 and will not return until Tue 11/27/2007. I will respond to your message when I return. ------------------------------ Message: 8 Date: Mon, 19 Nov 2007 15:15:02 -0600 From: "Ingles Claire" Subject: RE: SPAM-LOW: Re: [Histonet] no subject To: Message-ID: <08A0A863637F1349BBFD83A96B27A50A1200A2@uwhis-xchng3.uwhis.hosp.wisc.edu> Content-Type: text/plain; charset="iso-8859-1" I am! I'll even boycott Friday too! Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Douglas D Deltour Sent: Mon 11/19/2007 9:01 AM To: 'Emily Sours'; histonet@lists.utsouthwestern.edu Subject: RE: SPAM-LOW: Re: [Histonet] no subject I honor of this thread I declare Thursday November 22nd National Histology Strike Day! We will not report to work on this day to show our dissatisfaction. Who is with me? ------------------------------ Message: 9 Date: Mon, 19 Nov 2007 18:04:33 EST From: AGrobe2555@aol.com Subject: [Histonet] IHC for MMP2, MMP9, TIMP1 and TIMP2? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Hello all, I have a panel of tissues (Large and small intestine, liver, spleen, kidney, heart, esophagus, lung) as well as normal artery and vein tissue (all are from sheep) that we are using to test/optimize IHC for MMP2, MMP9, TIMP1 and TIMP2. We have stained for MMP9 so far, and see some staining in cells (appear to be macrophages) located in the small and large intestine, and in the adventitial/ablumenal tissue of the vein. We will be doing the other stains in the days to come. My difficulty is that I am in unknown territory with these antibodies. Has anyone of you stained for these before? What should I be looking for as a positive signal in these tissues? Can anyone point me to some reference images? In the interim, I will continue to look for information as the product sheets are not really providing information as to appropriate + control tissues. Thank you, Albert Albert C. Grobe, PhD International Heart Institute of Montana Foundation Tissue Engineering Lab, Saint Patrick Hospital ************************************** See what's new at http://www.aol.com ------------------------------ Message: 10 Date: Mon, 19 Nov 2007 19:33:31 -0500 From: "Lee & Peggy Wenk" Subject: RE: [Histonet] re:B-5 solution To: , Message-ID: <001201c82b0c$faad1c30$0202a8c0@HPPav2> Content-Type: text/plain; charset="us-ascii" Why not zinc formalin? Usually, if people have tried zinc formalin and didn't like it, it's because they didn't leave the tissue in the zinc formalin long enough. Usually they have left it for the same amount of time as the B5, and as a result the nuclear detail is lousy. The secret is that zinc salts binds a little slower than mercuric chloride. So take your B5 time, and multiply it by 1.5, and that will be a good starting point for zinc formalin. In other words, if the biopsies were fixed in B5 in 2 hours, then 2 x 1.5 = 3 hours would be a good guess at how long to fix in zinc formalin. Peggy A. 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 ATOJSU@aol.com Sent: Monday, November 19, 2007 2:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re:B-5 solution We have 1 Pathologist who insists on using B-5 solution for nuclear detail on small biopsies. How many labs are still using B 5 or what is you suggestion other than Zinc Formalin for fixation. Thanks. ************************************** See what's new at http://www.aol.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Tue, 20 Nov 2007 01:28:23 +0000 (GMT) From: kamal prasad Subject: [Histonet] (no subject) To: histonet@lists.utsouthwestern.edu Message-ID: <728317.58538.qm@web8705.mail.in.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Yes , 10% formic acid is best decalcifier it will keep cell morphology better. Formic acod need to mix it with 10%NBF . --------------------------------- Forgot the famous last words? Access your message archive online. Click here. ------------------------------ Message: 12 Date: Mon, 19 Nov 2007 17:37:40 -0800 From: "Goodpaster, Tracy A" Subject: RE: [Histonet] QIHC To: "Tarango, Mark" , Message-ID: Content-Type: text/plain; charset="us-ascii" I am also interested in this information. I don't know how specific it will be, especially over clinical lab regulations. Any additional information would be helpful! Thanks! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tarango, Mark Sent: Sunday, November 11, 2007 3:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] QIHC To anyone who has taken the QIHC exam, what kinds of questions does it ask? I've got a few books from the reading list, but I have no idea where to focus my attention. It is more about staining techniques or immunophenotyping? Quality control stuff? What's on the test that you might not expect? Is it a difficult exam? Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 "EMF " made the following annotations. ------------------------------------------------------------------------ ------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ======================================================================== ====== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Tue, 20 Nov 2007 11:28:27 +0200 From: "K M" Subject: [Histonet] Prognostic factors in colorectal cancers To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Dear colleagues I am doing study on IHC in colorectal cancers in human tissues and correlate them with prognosis .I used an antibody for Claudin-1(a tight junction protein found in cell membrane) as a prognostic factor.Ineed another prognostic factor for colorectal cancer for that I am enquiring if any one know other strong prognostic factor used in colorectal cancers? Dr.K.M.Omer ------------------------------ Message: 14 Date: Tue, 20 Nov 2007 06:08:56 -0600 From: "Joe Nocito" Subject: Re: [Histonet] forceps and bunsen burners To: "Cheryl R. Kerry" , Message-ID: <015d01c82b6e$207b2f40$0302a8c0@yourxhtr8hvc4p> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original the good ole days. The lab I'm at now used to used micro loop flamers. It was like playing the game "operation", if you touched the sides, sparks shoot up. It does cause a nice light show with the lights off. My other place of employment, we used alcohol burners, others used gauze. Personally, I miss the days of having a cigarette hanging from my mouth while cutting and coverslipping. I admit, I'm much healthier today, but miss the good ole days. JTT ----- Original Message ----- From: "Cheryl R. Kerry" To: Sent: Sunday, November 18, 2007 10:21 AM Subject: [Histonet] forceps and bunsen burners >I really miss Bunsen burners! Now we use a lot of kimwipes. > > Kemlo--I recall the days of wiping your embedding center with a xylene > soaked rag...me with a cuppa joe on my 'tome and others with cigarettes > hanging out the sides of their mouths...and yes, the occasional lab fires > that resulted!! And I really miss REAL light green SF yellowish! > > Cheryl > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of kemlo > Sent: Sunday, November 18, 2007 4:22 AM > To: 'sheila adey'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] (no subject) > > In the good old days of Bunsen burners we used to fry the end of the > forceps; nothing survived that and if it did it was readily identifiable. > Alas the Bunsen burner has been consigned to the politically incorrect as > the 'Scientists' of today would incinerate themselves whilst the > 'Technicians' of yesterday didn't (well not often). > > The sad demise of mercury, lead, Bunsen burners, formalin, anything too > hot, > too cold, too explosive, too poisonous, etc. Having your tea in the Lab > next > to the specimens and processing TB specimens 'on the bench'. > > Would Histology have the techniques and stains it now has if harmful > chemicals had not been experimented with? Will anything new be discovered > by > the HistoTech if all that we can use is 'safe' chemicals and procedures? I > don't see the kids fiddling with things like we use to, no explosions, no > fires and no injuries. Am I just an old reactionary waiting to be put out > to > pasture and ruminate on what was? > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of sheila > adey > Sent: 17 November 2007 20:41 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] (no subject) > > > Hi Netters, > > We are trying to minimize possible embedding contaminations. What are > other > people doing to prevent contamination due to forceps etc. > > Thanks in advanceSheila Adey HT MLTPort Huron HospitalMichigan > _________________________________________________________________ > Are you ready for Windows Live Messenger Beta 8.5 ? Get the latest for > free > today! > http://entertainment.sympatico.msn.ca/WindowsLiveMessenger__________________ > _____________________________ > Histonet mailing list > Histonet@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: 15 Date: Tue, 20 Nov 2007 08:08:36 -0800 From: Victor Tobias Subject: [Histonet] New RA Lamb Cassette Printer UK Only To: HISTONET Message-ID: <47430684.4000608@pathology.washington.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Has anyone purchased or seen this new cassette labeler? We are still waiting for Thermo to release the US version. I have heard it will print cassettes with bar codes in 2.5 seconds, which is remarkable. Victor -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. ------------------------------ Message: 16 Date: 20 Nov 2007 11:48:11 -0500 From: Tara Ollis Subject: [Histonet] Histologist Job in ATL GA To: histonet@lists.utsouthwestern.edu Message-ID: <18512542.1195577290900.JavaMail.cfservice@webserver29> Content-Type: text/plain; charset="utf-8" Hello- I have a Histologist position available in a dermatologist office in the greater Atlanta area. They need someone with MOHS experience and prefer HTL Certification. Feel free to email me your resume or call me for more details. Thanks! Tara Ollis | TeamStaff Rx, Inc. Office: 1.877.523.9897 ext.5511 | www.teamstaffrx.com ------------------------------ Message: 17 Date: Tue, 20 Nov 2007 11:54:45 -0600 From: MVaughan4@ucok.edu Subject: [Histonet] Matrigel as culture medium - try agarose embedding To: histonet@lists.utsouthwestern.edu Cc: TJJ@Stowers-Institute.org Message-ID: Content-Type: text/plain; charset="US-ASCII" Teri, Can you embed or surround the matrigel mixture in a low-melting agarose first, then postfix the agarose/matrigel mixture in formalin, followed by paraffin embedding? I have used it with collagen matrices. Mel Melville B. Vaughan, Ph. D. Assistant Professor of Biology Campus Coordinator, NSF Sure-Step program University of Central Oklahoma 100 N. University Drive Edmond, OK 73034 http://www.biology.ucok.edu/PersonalPages/mvaughan/Default.htm Message: 8 Date: Fri, 16 Nov 2007 14:42:54 -0600 From: "Keller, Charles" Subject: RE: [Histonet] Matrigel as culture medium To: "Johnson, Teri" , Message-ID: Content-Type: text/plain; charset="us-ascii" Dear Teri, a possible solution is to use Extracel (from Glycosan Inc in Utah) instead of Matrigel. I've fixed Extracel in 10% formalin and sectioned in in paraffin. It has the advantage of being synthetic (Matrigel has viral contamination from time to time). Sincerely, Charles -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Friday, November 16, 2007 9:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Matrigel as culture medium One of our researchers is using matrigel as a culture medium on chambered slides. We tried paraffin embedding them, and found the Matrigel wasn't really solid enough to hold the cellular structure in place. We found it particularly difficult to keep some sort of consistency scraping it off (it had the consistency of warm jelly). I'd love to hear some ideas on how we can accomplish sample processing without centrifugation. Ideally it would be good to encase the cells in a matrix that would solidfy enough to embed as a cell block, keeping the cellular structure intact. Thanks for your help! Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------- **CONFIDENTIALITY** -This email (including any attachments) may contain confidential, proprietary and privileged information. Any unauthorized disclosure or use of this information is prohibited. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 28 **************************************** From jstaruk <@t> masshistology.com Tue Nov 20 18:49:17 2007 From: jstaruk <@t> masshistology.com (jstaruk) Date: Tue Nov 20 18:50:08 2007 Subject: [Histonet] How to keep cryostating without getting frostbite? In-Reply-To: <0ML2xA-1Iuao51g7u-0005Ac@mrelayeu.kundenserver.de> Message-ID: Hey all, Are there any tricks on how to keep your fingers from getting frostbite while doing cryostat sectioning for 8 hours a day? My poor techs who have been cutting frozen sections for several straight days now are starting to complain about these inhumane conditions! Do all of you MOHS techs have bleeding, chapped hands and numb fingertips? Thank you in advance for any suggestions and Happy Thanksgiving to all! Jim _____________________ Jim Staruk Mass Histology Service www.masshistology.com From louise.renton <@t> gmail.com Wed Nov 21 04:15:20 2007 From: louise.renton <@t> gmail.com (louise renton) Date: Wed Nov 21 04:22:22 2007 Subject: [Histonet] how long does it take? Message-ID: Dear Histonetters, First off, let me say what a great experience the NSH symposium was! It was great to finally meet y'all. Made me realise that I am not the only one suffering! So here's my question rearding IHC....... What is a reasonable work-up time for a new antibody? The parameters are: 1. Ab researh only, not used in the particular species of animal I am using 2. Ab titre needs to be assessed 3. not sure what a good control might be 4.Will be using archival tissue where fixation/processing may be questionable 5. using a manual ABC/DAB technique 6. All buffers etc made up in-house Any ideas?? Best regards -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "There are nights when the wolves are silent and only the moon howls". George Carlin No trees were killed in the sending of this message. However, many electrons were terribly inconvenienced. From daliaelrouby <@t> hotmail.com Wed Nov 21 04:36:43 2007 From: daliaelrouby <@t> hotmail.com (Dalia El Rouby) Date: Wed Nov 21 04:36:48 2007 Subject: [Histonet] (no subject) Message-ID: Dear collegues: could someone give me the detailed procedure of mercuric bromephenol blue staining for protein in paraffin sections. Best regrds Dalia El-Rouby _________________________________________________________________ Invite your mail contacts to join your friends list with Windows Live Spaces. It's easy! http://spaces.live.com/spacesapi.aspx?wx_action=create&wx_url=/friends.aspx&mkt=en-us From rjbuesa <@t> yahoo.com Wed Nov 21 07:16:47 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 21 07:16:51 2007 Subject: [Histonet] how long does it take? In-Reply-To: Message-ID: <2025.80966.qm@web61219.mail.yahoo.com> Louise: If that is the only thing you are dedicated to doing, you will have to make a literature search to find out how other people have handled this Ab: that would be one week, during which you can also locate archival tissues to test based in what has been done. If there is no published information this time will be reduced and added to the experimental phase. Developing your "checker-board" of dilutions vs. incubation period, would take another week. Preparing your reagents can be done simultaneously with any others you have to prepare daily. Finally you will have to analyze your results, select the best dilution, and make some "final adjustments"; that would be a third week. So 3 weeks if that is the only thing you are doing (about 6h x 5 days x 3 weeks = 90 hours), BUT if you work less than 6 h/day in the project, extend the time to get to your 90 hours. That is how I see the time neded for this project! Ren? J. louise renton wrote: Dear Histonetters, First off, let me say what a great experience the NSH symposium was! It was great to finally meet y'all. Made me realise that I am not the only one suffering! So here's my question rearding IHC....... What is a reasonable work-up time for a new antibody? The parameters are: 1. Ab researh only, not used in the particular species of animal I am using 2. Ab titre needs to be assessed 3. not sure what a good control might be 4.Will be using archival tissue where fixation/processing may be questionable 5. using a manual ABC/DAB technique 6. All buffers etc made up in-house Any ideas?? Best regards -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "There are nights when the wolves are silent and only the moon howls". George Carlin No trees were killed in the sending of this message. However, many electrons were terribly inconvenienced. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From petepath <@t> yahoo.com Wed Nov 21 07:42:39 2007 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Wed Nov 21 07:49:27 2007 Subject: [Histonet] How to keep cryostating without getting frostbite? Message-ID: <234908.36292.qm@web45114.mail.sp1.yahoo.com> I recently suggested to a cryostat manufacturer that the route the exhaust from the motor through a hand warming device on the side of the cryostat. It was not taken seriously. I thought it was a good idea. Are there any very thin cotton gloves that could be warn under rubber gloves? How about double gloving? How about a warm puppy next to the cryostat! Happy Thanksgiving Histonetters! 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 b-frederick <@t> northwestern.edu Wed Nov 21 07:55:45 2007 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Wed Nov 21 07:55:57 2007 Subject: [Histonet] how long does it take? In-Reply-To: Message-ID: <000c01c82c46$3a013670$d00f7ca5@lurie.northwestern.edu> Louise, We tell researchers a minimum of two weeks for any project involving IHC since we have multiple clients- 1 week longer if titreing is involved. Some times you get it right away,sometimes not. Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise renton Sent: Wednesday, November 21, 2007 4:15 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] how long does it take? Dear Histonetters, First off, let me say what a great experience the NSH symposium was! It was great to finally meet y'all. Made me realise that I am not the only one suffering! So here's my question rearding IHC....... What is a reasonable work-up time for a new antibody? The parameters are: 1. Ab researh only, not used in the particular species of animal I am using 2. Ab titre needs to be assessed 3. not sure what a good control might be 4.Will be using archival tissue where fixation/processing may be questionable 5. using a manual ABC/DAB technique 6. All buffers etc made up in-house Any ideas?? Best regards -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "There are nights when the wolves are silent and only the moon howls". George Carlin No trees were killed in the sending of this message. However, many electrons were terribly inconvenienced. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From oshel1pe <@t> cmich.edu Wed Nov 21 07:58:20 2007 From: oshel1pe <@t> cmich.edu (Philip Oshel) Date: Wed Nov 21 07:58:33 2007 Subject: [Histonet] How to keep cryostating without getting frostbite? In-Reply-To: <234908.36292.qm@web45114.mail.sp1.yahoo.com> References: <234908.36292.qm@web45114.mail.sp1.yahoo.com> Message-ID: I'd prefer a Himalayan myself -- much fluffier and warmer. Would be static problems, though. Gloves: polypropylene glove liners. Fit snuggly so one can still work with them, and do a decent enough job of protecting fingers when handling very cold surfaces. Worked for me in the Arctic and Antarctic, although I haven't used them yet in a cryostat. Phil >I recently suggested to a cryostat manufacturer that the route the >exhaust from the > motor through a hand warming device on the side of the cryostat. It was not > taken seriously. I thought it was a good idea. Are there any very >thin cotton gloves that > could be warn under rubber gloves? How about double gloving? How >about a warm puppy next to the cryostat! Happy Thanksgiving >Histonetters! > > 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 -- Philip Oshel Microscopy Facility Supervisor Biology Department 024C Brooks Hall Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 From am3309 <@t> uga.edu Wed Nov 21 09:13:11 2007 From: am3309 <@t> uga.edu (Abigail M. Butler) Date: Wed Nov 21 09:13:16 2007 Subject: [Histonet] Histogel Question Message-ID: <20071121101311.JJD02650@punts1.cc.uga.edu> I completely understand how to use histogel to embed free floating cells. But how would I embed a piece of tissue that obviously is not placed in a tube? I am considering purchasing Histogel for the first time and have many ideas of how I could use it, just not sure the proper steps to take to embed let's say two mice eyes. Thanks Abbie Butler, HT (ASCP), QIHC University of Georgia College of Veterinary Medicine From jennifer.l.hofecker <@t> Vanderbilt.Edu Wed Nov 21 09:20:04 2007 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Wed Nov 21 09:20:13 2007 Subject: [Histonet] Histogel Question In-Reply-To: <20071121101311.JJD02650@punts1.cc.uga.edu> Message-ID: <898D946569A27444B65667A49C074052013D7525@mailbe06.mc.vanderbilt.edu> Hi Abby. First, Histogel is great! We published a paper in the Journal of Histotechnology, Neuropath special issue, in June. It deals with using Histogel for CNS samples, but can totally be used as a reference for other tissues. For tissues (not cell suspensions) I recommend placing the tissue in a Peel Away mold, squirt molten histogel on the tissue and cool the block (refrig). We have completely eliminated the use of "T-bags" for minute specimens. For larger fragments of brain tumor, we use a larger base mold and just add a little histogel to "hold the fragments" together. It will reduce tissue shrinkage and will preserve the orientation. This is the reader's digest version. If you have additional questions, don't hesitate to call or email me. Have a great day. Jennifer Jennifer L. Hofecker, HT(ASCP) Vanderbilt University Medical Center Division of Neuropathology Nashville, TN ph. (615)343-0083 fax. (615)343-7089 NSH Quality Control Committee Chair -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Abigail M. Butler Sent: Wednesday, November 21, 2007 9:13 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histogel Question I completely understand how to use histogel to embed free floating cells. But how would I embed a piece of tissue that obviously is not placed in a tube? I am considering purchasing Histogel for the first time and have many ideas of how I could use it, just not sure the proper steps to take to embed let's say two mice eyes. Thanks Abbie Butler, HT (ASCP), QIHC University of Georgia College of Veterinary Medicine _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Mikeykn <@t> aol.com Wed Nov 21 09:59:33 2007 From: Mikeykn <@t> aol.com (Mikeykn@aol.com) Date: Wed Nov 21 09:59:41 2007 Subject: [Histonet] job opening memphis,tn Message-ID: Registered Histotech Full time M-F with Saturday rotations (hours variable) Exp. in IHC, IF,General and biopsy processing, embedding,special staining, microtomy, etc Competative pay and excellent benefits. Great place to work. Interested ? Send resume to Human Resources Trumbull Laboratories, LLC 6046 Knight Arnold Rd. St.101 Memphis, Tn. 38115 **************************************Check out AOL's list of 2007's hottest products. (http://money.aol.com/special/hot-products-2007?NCID=aoltop00030000000001) From tkngflght <@t> yahoo.com Wed Nov 21 10:15:11 2007 From: tkngflght <@t> yahoo.com (Cheryl R. Kerry) Date: Wed Nov 21 10:15:45 2007 Subject: [Histonet] How to keep cryostating without getting frostbite? In-Reply-To: Message-ID: <011f01c82c59$b18e1790$6901a8c0@CHERYLSLAPTOP> Glove liners!! You can get these by the box at sporting goods stores--cotton glove liners that give you a little insulation between the latex or other sorts of biological protective gloves and the cold. Also, a tap of lukewarm running water over the gloves to warm them between cases---your hands don't get wet but they warm up. BE CAREFUL the water isn't too warm--it's the difference in temperature that can cause a burn, not just the temp of the water itself. Hope this helps--used to cut monkey brains for 10 hrs a day for weeks on end...the liners and a good jazz radio station kept me pretty happy in my work! Cheryl Cheryl R. Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab, one great tech at a time. 281.852.9457 office 281.883.7704 cell 800.756.3309 fax and alternate phone -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of jstaruk Sent: Tuesday, November 20, 2007 6:49 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] How to keep cryostating without getting frostbite? Hey all, Are there any tricks on how to keep your fingers from getting frostbite while doing cryostat sectioning for 8 hours a day? My poor techs who have been cutting frozen sections for several straight days now are starting to complain about these inhumane conditions! Do all of you MOHS techs have bleeding, chapped hands and numb fingertips? Thank you in advance for any suggestions and Happy Thanksgiving to all! Jim _____________________ Jim Staruk Mass Histology Service www.masshistology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Wed Nov 21 10:16:42 2007 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Wed Nov 21 10:16:53 2007 Subject: [Histonet] How to keep cryostating without getting frostbite? In-Reply-To: Message-ID: <4EBFF65383B74D49995298C4976D1D5E273CFA@LSRIEXCH1.lsmaster.lifespan.org> Just having your hands in the cold air isn't usually the problem. Each tech, according to their particular style, usually rests their hand on the knife holder in some manner for stability while picking up the sections on the slides. The rate of extraction of heat by contact with -20 degree metal is much greater than the rate by simple exposure to -20 degree air. It is these points of repeated contact between skin and metal that usually suffer from frostbite. I have tried various kinds of gloves and slip-on finger protectors, but in the final analysis I find that bandaids carefully applied to the specific points of contact provide enough insulation to prevent skin damage. I buy the extra wide ones. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of jstaruk > Sent: Tuesday, November 20, 2007 7:49 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] How to keep cryostating without getting frostbite? > > Hey all, > > Are there any tricks on how to keep your fingers from getting frostbite > while doing cryostat sectioning for 8 hours a day? My poor techs who have > been cutting frozen sections for several straight days now are starting to > complain about these inhumane conditions! Do all of you MOHS techs have > bleeding, chapped hands and numb fingertips? > > Thank you in advance for any suggestions and Happy Thanksgiving to all! > > Jim > > _____________________ > Jim Staruk > Mass Histology Service > www.masshistology.com > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From settembr <@t> umdnj.edu Wed Nov 21 10:37:44 2007 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Wed Nov 21 10:38:16 2007 Subject: [Histonet] Histogel Question Message-ID: Abbie, I have used Histogel to embed very small human eye biopsies. What do you mean, "that obviously is not placed in a tube?"? We pick up the small specimen carefully with forceps and place the tissue at the very bottom of the gel and when it goes into a cassette, it is placed facing down. We make sure that the embedding histotech understands exactly where it is for processing and embedding. Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ USA >>> "Abigail M. Butler" 11/21/07 10:13 AM >>> I completely understand how to use histogel to embed free floating cells. But how would I embed a piece of tissue that obviously is not placed in a tube? I am considering purchasing Histogel for the first time and have many ideas of how I could use it, just not sure the proper steps to take to embed let's say two mice eyes. Thanks Abbie Butler, HT (ASCP), QIHC University of Georgia College of Veterinary Medicine _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From algranth <@t> u.arizona.edu Wed Nov 21 11:22:36 2007 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Wed Nov 21 11:28:06 2007 Subject: [Histonet] How to keep cryostating without getting frostbite? In-Reply-To: <4EBFF65383B74D49995298C4976D1D5E273CFA@LSRIEXCH1.lsmaster. lifespan.org> References: <4EBFF65383B74D49995298C4976D1D5E273CFA@LSRIEXCH1.lsmaster.lifespan.org> Message-ID: <6.2.3.4.1.20071121100954.01faafa8@algranth.inbox.email.arizona.edu> I cut little rectangular pieces of styrofoam from any of the cartons that are shipped in with cold or frozen items and "glue" them to each side of the knife holder with OCT. This provides a great area to rest my hands while having to manipulate a frozen section. The styrofoam doesn't get cold and it is a comfty place to put your hands and also a slide if you have to pick up multiple sections on one slide. The tissue and OCT doesn't thaw or the slide doesn't get too cold for another section to attach. This excellent piece of advice came from Gayle Callis - thank you again Gayle!!! PS: When I want to clean out the cryostat I just turn it off and take out the knife holder and when the OCT warms up the styrofoam comes off and can be reattached when the cryostat is all clean and ready to be turned back on. I was reading where somebody suggested something to a manufacturer and the idea was not readily accepted and I likewise have made what I thought were excellent suggestions and was met with similar responses - this styrofoam thing was just one of them. Andi At 09:16 AM 11/21/2007, Monfils, Paul wrote: >Just having your hands in the cold air isn't usually the problem. >Each tech, according to their particular style, usually rests their >hand on the knife holder in some manner for stability while picking >up the sections on the slides. The rate of extraction of heat by >contact with -20 degree metal is much greater than the rate by >simple exposure to -20 degree air. It is these points of repeated >contact between skin and metal that usually suffer from >frostbite. I have tried various kinds of gloves and slip-on finger >protectors, but in the final analysis I find that bandaids carefully >applied to the specific points of contact provide enough insulation >to prevent skin damage. I buy the extra wide ones. > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf > of jstaruk > > Sent: Tuesday, November 20, 2007 7:49 PM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] How to keep cryostating without getting frostbite? > > > > Hey all, > > > > Are there any tricks on how to keep your fingers from getting frostbite > > while doing cryostat sectioning for 8 hours a day? My poor techs who have > > been cutting frozen sections for several straight days now are starting to > > complain about these inhumane conditions! Do all of you MOHS techs have > > bleeding, chapped hands and numb fingertips? > > > > Thank you in advance for any suggestions and Happy Thanksgiving to all! > > > > Jim > > > > _____________________ > > Jim Staruk > > Mass Histology Service > > www.masshistology.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 ..................................................................... : 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 am3309 <@t> uga.edu Wed Nov 21 11:28:14 2007 From: am3309 <@t> uga.edu (Abigail M. Butler) Date: Wed Nov 21 11:28:20 2007 Subject: [Histonet] RE: Anti-dog IgG Message-ID: <20071121122814.JJF06401@punts1.cc.uga.edu> MaryAnn, I use only anti-dog IgG.....and my dilution is 1:150,000. Crazy, isn't it? I also used HIER for pretreatment with citrate buffer. There is not much background. Hope this helps Abbie Butler, HT (ASCP), QIHC University of Georgia College of Veterinary Medicine From PMonfils <@t> Lifespan.org Wed Nov 21 11:36:05 2007 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Wed Nov 21 11:36:16 2007 Subject: [Histonet] -80 degree freezer, repair question Message-ID: <4EBFF65383B74D49995298C4976D1D5E273CFC@LSRIEXCH1.lsmaster.lifespan.org> We have a rather old Baxter Scientific Cryo-Fridge chest type -80 degree freezer that works fine except the lid gasket is shot, so cold air leaks out and frost and ice build up around the edges of the lid. Does anyone know of someone who could replace this gasket? Or somewhere we could purchase the part to install ourselves? We are also unable to find particle filters for the front door of the compressor compartment. From gu.lang <@t> gmx.at Wed Nov 21 11:50:42 2007 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Wed Nov 21 11:50:50 2007 Subject: [Histonet] frozens from lung-tissue Message-ID: <000601c82c67$09d1b0d0$6412a8c0@dielangs.at> Hi to all, I need the help of those, who have experience on making good frozens from lung-tissue. We look for a method for freezing lung-tissue without the usage of liquid nitrogen, co2 or isopentan. - Just on the cryocut. The head of the institute is not satisfied with the quality and wants us to do better. Our usual method is to make a OCT-layer on a chuck, let it freeze, give the tissue on it, surround with OCT, put the cold weight on it, let it freeze and make the sections. We tried putting the tissue in the liquid OCT on the chuck without a layer and freezing it in a mold with the chuck on top. I also tried first freezing the piece solely in liquid nitrogen or isopentan, then putting it on a chuck. All in all - my boss wasn't happy with it. Any input will be appreciated. Thank you in advance Gudrun Lang From tkngflght <@t> yahoo.com Wed Nov 21 12:11:41 2007 From: tkngflght <@t> yahoo.com (Cheryl R. Kerry) Date: Wed Nov 21 12:12:19 2007 Subject: [Histonet] frozens from lung-tissue In-Reply-To: <000601c82c67$09d1b0d0$6412a8c0@dielangs.at> Message-ID: <014301c82c69$f83573e0$6901a8c0@CHERYLSLAPTOP> Gudrun--I suspect this isn't quite what you were looking for but have you guys tried a cryobath? It's a little blue countertop 'freezer' unit that has well of pre-chilled isopentane in it, always cold enough to snap freeze without the drama of liquid nitrogen. It is about the size of a big toaster and is ready to go 24/7 with minimal maintenance (temp chart and periodic cleaning). It wasn't overly expensive from what I remember for the great quality of the end results. We mounted the tissue on a pre-frozen OCT-covered chuck, covered it with liquid OCT, lowered it into the cryobath for about 30-40 second and cut...beautiful, artifact free, duplicable and fast. What are the reasons s/he doesn't want the more standard snap freeze methods? Is it a time or expense issue? Cheryl -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Wednesday, November 21, 2007 11:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] frozens from lung-tissue Hi to all, I need the help of those, who have experience on making good frozens from lung-tissue. We look for a method for freezing lung-tissue without the usage of liquid nitrogen, co2 or isopentan. - Just on the cryocut. The head of the institute is not satisfied with the quality and wants us to do better. Our usual method is to make a OCT-layer on a chuck, let it freeze, give the tissue on it, surround with OCT, put the cold weight on it, let it freeze and make the sections. We tried putting the tissue in the liquid OCT on the chuck without a layer and freezing it in a mold with the chuck on top. I also tried first freezing the piece solely in liquid nitrogen or isopentan, then putting it on a chuck. All in all - my boss wasn't happy with it. Any input will be appreciated. Thank you in advance Gudrun Lang From JeromeS <@t> rice.edu Wed Nov 21 12:11:42 2007 From: JeromeS <@t> rice.edu (J. Saltarrelli) Date: Wed Nov 21 12:12:22 2007 Subject: [Histonet] unsubscribe In-Reply-To: <20071121180648.8833F384FB7@mh1.mail.rice.edu> References: <20071121180648.8833F384FB7@mh1.mail.rice.edu> Message-ID: Please unsubscribe me -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, November 21, 2007 12:07 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 48, Issue 30 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Re: Histogel Question (Dana Settembre) 2. RE: How to keep cryostating without getting frostbite? (Andrea Grantham) 3. RE: Anti-dog IgG (Abigail M. Butler) 4. -80 degree freezer, repair question (Monfils, Paul) 5. frozens from lung-tissue (Gudrun Lang) ---------------------------------------------------------------------- Message: 1 Date: Wed, 21 Nov 2007 11:37:44 -0500 From: "Dana Settembre" Subject: Re: [Histonet] Histogel Question To: , "Abigail M. Butler" Message-ID: Content-Type: text/plain; charset=US-ASCII Abbie, I have used Histogel to embed very small human eye biopsies. What do you mean, "that obviously is not placed in a tube?"? We pick up the small specimen carefully with forceps and place the tissue at the very bottom of the gel and when it goes into a cassette, it is placed facing down. We make sure that the embedding histotech understands exactly where it is for processing and embedding. Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ USA >>> "Abigail M. Butler" 11/21/07 10:13 AM >>> I completely understand how to use histogel to embed free floating cells. But how would I embed a piece of tissue that obviously is not placed in a tube? I am considering purchasing Histogel for the first time and have many ideas of how I could use it, just not sure the proper steps to take to embed let's say two mice eyes. Thanks Abbie Butler, HT (ASCP), QIHC University of Georgia College of Veterinary Medicine _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Wed, 21 Nov 2007 10:22:36 -0700 From: Andrea Grantham Subject: RE: [Histonet] How to keep cryostating without getting frostbite? To: "Monfils, Paul" , Message-ID: <6.2.3.4.1.20071121100954.01faafa8@algranth.inbox.email.arizona.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I cut little rectangular pieces of styrofoam from any of the cartons that are shipped in with cold or frozen items and "glue" them to each side of the knife holder with OCT. This provides a great area to rest my hands while having to manipulate a frozen section. The styrofoam doesn't get cold and it is a comfty place to put your hands and also a slide if you have to pick up multiple sections on one slide. The tissue and OCT doesn't thaw or the slide doesn't get too cold for another section to attach. This excellent piece of advice came from Gayle Callis - thank you again Gayle!!! PS: When I want to clean out the cryostat I just turn it off and take out the knife holder and when the OCT warms up the styrofoam comes off and can be reattached when the cryostat is all clean and ready to be turned back on. I was reading where somebody suggested something to a manufacturer and the idea was not readily accepted and I likewise have made what I thought were excellent suggestions and was met with similar responses - this styrofoam thing was just one of them. Andi At 09:16 AM 11/21/2007, Monfils, Paul wrote: >Just having your hands in the cold air isn't usually the problem. >Each tech, according to their particular style, usually rests their >hand on the knife holder in some manner for stability while picking >up the sections on the slides. The rate of extraction of heat by >contact with -20 degree metal is much greater than the rate by >simple exposure to -20 degree air. It is these points of repeated >contact between skin and metal that usually suffer from >frostbite. I have tried various kinds of gloves and slip-on finger >protectors, but in the final analysis I find that bandaids carefully >applied to the specific points of contact provide enough insulation >to prevent skin damage. I buy the extra wide ones. > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf > of jstaruk > > Sent: Tuesday, November 20, 2007 7:49 PM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] How to keep cryostating without getting frostbite? > > > > Hey all, > > > > Are there any tricks on how to keep your fingers from getting frostbite > > while doing cryostat sectioning for 8 hours a day? My poor techs who have > > been cutting frozen sections for several straight days now are starting to > > complain about these inhumane conditions! Do all of you MOHS techs have > > bleeding, chapped hands and numb fingertips? > > > > Thank you in advance for any suggestions and Happy Thanksgiving to all! > > > > Jim > > > > _____________________ > > Jim Staruk > > Mass Histology Service > > www.masshistology.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 ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html ------------------------------ Message: 3 Date: Wed, 21 Nov 2007 12:28:14 -0500 (EST) From: "Abigail M. Butler" Subject: [Histonet] RE: Anti-dog IgG To: histonet@lists.utsouthwestern.edu Message-ID: <20071121122814.JJF06401@punts1.cc.uga.edu> Content-Type: text/plain; charset=us-ascii MaryAnn, I use only anti-dog IgG.....and my dilution is 1:150,000. Crazy, isn't it? I also used HIER for pretreatment with citrate buffer. There is not much background. Hope this helps Abbie Butler, HT (ASCP), QIHC University of Georgia College of Veterinary Medicine ------------------------------ Message: 4 Date: Wed, 21 Nov 2007 12:36:05 -0500 From: "Monfils, Paul" Subject: [Histonet] -80 degree freezer, repair question To: Message-ID: <4EBFF65383B74D49995298C4976D1D5E273CFC@LSRIEXCH1.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" We have a rather old Baxter Scientific Cryo-Fridge chest type -80 degree freezer that works fine except the lid gasket is shot, so cold air leaks out and frost and ice build up around the edges of the lid. Does anyone know of someone who could replace this gasket? Or somewhere we could purchase the part to install ourselves? We are also unable to find particle filters for the front door of the compressor compartment. ------------------------------ Message: 5 Date: Wed, 21 Nov 2007 18:50:42 +0100 From: "Gudrun Lang" Subject: [Histonet] frozens from lung-tissue To: Message-ID: <000601c82c67$09d1b0d0$6412a8c0@dielangs.at> Content-Type: text/plain; charset="us-ascii" Hi to all, I need the help of those, who have experience on making good frozens from lung-tissue. We look for a method for freezing lung-tissue without the usage of liquid nitrogen, co2 or isopentan. - Just on the cryocut. The head of the institute is not satisfied with the quality and wants us to do better. Our usual method is to make a OCT-layer on a chuck, let it freeze, give the tissue on it, surround with OCT, put the cold weight on it, let it freeze and make the sections. We tried putting the tissue in the liquid OCT on the chuck without a layer and freezing it in a mold with the chuck on top. I also tried first freezing the piece solely in liquid nitrogen or isopentan, then putting it on a chuck. All in all - my boss wasn't happy with it. Any input will be appreciated. Thank you in advance Gudrun Lang ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 30 **************************************** From pierre.chaumat <@t> alphelys.com Wed Nov 21 12:31:39 2007 From: pierre.chaumat <@t> alphelys.com (Pierre CHAUMAT) Date: Wed Nov 21 12:31:46 2007 Subject: [Histonet] RE: Histonet Digest, Vol 48, Issue 30 In-Reply-To: <0ML4Q0-1IuttH3vPp-0007gU@mx.kundenserver.de> Message-ID: <0ML25U-1IuuMM24Ch-00047y@mrelayeu.kundenserver.de> Dear Gudrun, I know using isopentane without a dedicated machine is somehow acrobatic and difficult. Moreover, maintaining the right temp is also a tricky exercise. We have developped a system calles SnapFrost that uses electrical compressors to cool down to -80?C an isopentane bath that will provide you with ultra-reproducible temperature for always high quality frozens. A path from Z?rich Univ Hosp has two of these units used in routine since a while. He has shown very good morphology and always much better than freezing bar of a cryostat. Additionally, it takes only few seconds to get there when a cryostat bar will take few minutes and time always counts especially in frozen diagnosis. Kind regards Pierre Your message---------------------------- Message: 5 Date: Wed, 21 Nov 2007 18:50:42 +0100 From: "Gudrun Lang" Subject: [Histonet] frozens from lung-tissue To: Message-ID: <000601c82c67$09d1b0d0$6412a8c0@dielangs.at> Content-Type: text/plain; charset="us-ascii" Hi to all, I need the help of those, who have experience on making good frozens from lung-tissue. We look for a method for freezing lung-tissue without the usage of liquid nitrogen, co2 or isopentan. - Just on the cryocut. The head of the institute is not satisfied with the quality and wants us to do better. Our usual method is to make a OCT-layer on a chuck, let it freeze, give the tissue on it, surround with OCT, put the cold weight on it, let it freeze and make the sections. We tried putting the tissue in the liquid OCT on the chuck without a layer and freezing it in a mold with the chuck on top. I also tried first freezing the piece solely in liquid nitrogen or isopentan, then putting it on a chuck. All in all - my boss wasn't happy with it. Any input will be appreciated. Thank you in advance Gudrun Lang -----Message d'origine----- De : histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] De la part de histonet-request@lists.utsouthwestern.edu Envoy? : mercredi 21 novembre 2007 19:02 ? : histonet@lists.utsouthwestern.edu Objet : Histonet Digest, Vol 48, Issue 30 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Re: Histogel Question (Dana Settembre) 2. RE: How to keep cryostating without getting frostbite? (Andrea Grantham) 3. RE: Anti-dog IgG (Abigail M. Butler) 4. -80 degree freezer, repair question (Monfils, Paul) 5. frozens from lung-tissue (Gudrun Lang) ---------------------------------------------------------------------- Message: 1 Date: Wed, 21 Nov 2007 11:37:44 -0500 From: "Dana Settembre" Subject: Re: [Histonet] Histogel Question To: , "Abigail M. Butler" Message-ID: Content-Type: text/plain; charset=US-ASCII Abbie, I have used Histogel to embed very small human eye biopsies. What do you mean, "that obviously is not placed in a tube?"? We pick up the small specimen carefully with forceps and place the tissue at the very bottom of the gel and when it goes into a cassette, it is placed facing down. We make sure that the embedding histotech understands exactly where it is for processing and embedding. Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ USA >>> "Abigail M. Butler" 11/21/07 10:13 AM >>> I completely understand how to use histogel to embed free floating cells. But how would I embed a piece of tissue that obviously is not placed in a tube? I am considering purchasing Histogel for the first time and have many ideas of how I could use it, just not sure the proper steps to take to embed let's say two mice eyes. Thanks Abbie Butler, HT (ASCP), QIHC University of Georgia College of Veterinary Medicine _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Wed, 21 Nov 2007 10:22:36 -0700 From: Andrea Grantham Subject: RE: [Histonet] How to keep cryostating without getting frostbite? To: "Monfils, Paul" , Message-ID: <6.2.3.4.1.20071121100954.01faafa8@algranth.inbox.email.arizona.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I cut little rectangular pieces of styrofoam from any of the cartons that are shipped in with cold or frozen items and "glue" them to each side of the knife holder with OCT. This provides a great area to rest my hands while having to manipulate a frozen section. The styrofoam doesn't get cold and it is a comfty place to put your hands and also a slide if you have to pick up multiple sections on one slide. The tissue and OCT doesn't thaw or the slide doesn't get too cold for another section to attach. This excellent piece of advice came from Gayle Callis - thank you again Gayle!!! PS: When I want to clean out the cryostat I just turn it off and take out the knife holder and when the OCT warms up the styrofoam comes off and can be reattached when the cryostat is all clean and ready to be turned back on. I was reading where somebody suggested something to a manufacturer and the idea was not readily accepted and I likewise have made what I thought were excellent suggestions and was met with similar responses - this styrofoam thing was just one of them. Andi At 09:16 AM 11/21/2007, Monfils, Paul wrote: >Just having your hands in the cold air isn't usually the problem. >Each tech, according to their particular style, usually rests their >hand on the knife holder in some manner for stability while picking >up the sections on the slides. The rate of extraction of heat by >contact with -20 degree metal is much greater than the rate by >simple exposure to -20 degree air. It is these points of repeated >contact between skin and metal that usually suffer from >frostbite. I have tried various kinds of gloves and slip-on finger >protectors, but in the final analysis I find that bandaids carefully >applied to the specific points of contact provide enough insulation >to prevent skin damage. I buy the extra wide ones. > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf > of jstaruk > > Sent: Tuesday, November 20, 2007 7:49 PM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] How to keep cryostating without getting frostbite? > > > > Hey all, > > > > Are there any tricks on how to keep your fingers from getting frostbite > > while doing cryostat sectioning for 8 hours a day? My poor techs who have > > been cutting frozen sections for several straight days now are starting to > > complain about these inhumane conditions! Do all of you MOHS techs have > > bleeding, chapped hands and numb fingertips? > > > > Thank you in advance for any suggestions and Happy Thanksgiving to all! > > > > Jim > > > > _____________________ > > Jim Staruk > > Mass Histology Service > > www.masshistology.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 ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html ------------------------------ Message: 3 Date: Wed, 21 Nov 2007 12:28:14 -0500 (EST) From: "Abigail M. Butler" Subject: [Histonet] RE: Anti-dog IgG To: histonet@lists.utsouthwestern.edu Message-ID: <20071121122814.JJF06401@punts1.cc.uga.edu> Content-Type: text/plain; charset=us-ascii MaryAnn, I use only anti-dog IgG.....and my dilution is 1:150,000. Crazy, isn't it? I also used HIER for pretreatment with citrate buffer. There is not much background. Hope this helps Abbie Butler, HT (ASCP), QIHC University of Georgia College of Veterinary Medicine ------------------------------ Message: 4 Date: Wed, 21 Nov 2007 12:36:05 -0500 From: "Monfils, Paul" Subject: [Histonet] -80 degree freezer, repair question To: Message-ID: <4EBFF65383B74D49995298C4976D1D5E273CFC@LSRIEXCH1.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" We have a rather old Baxter Scientific Cryo-Fridge chest type -80 degree freezer that works fine except the lid gasket is shot, so cold air leaks out and frost and ice build up around the edges of the lid. Does anyone know of someone who could replace this gasket? Or somewhere we could purchase the part to install ourselves? We are also unable to find particle filters for the front door of the compressor compartment. ------------------------------ Message: 5 Date: Wed, 21 Nov 2007 18:50:42 +0100 From: "Gudrun Lang" Subject: [Histonet] frozens from lung-tissue To: Message-ID: <000601c82c67$09d1b0d0$6412a8c0@dielangs.at> Content-Type: text/plain; charset="us-ascii" Hi to all, I need the help of those, who have experience on making good frozens from lung-tissue. We look for a method for freezing lung-tissue without the usage of liquid nitrogen, co2 or isopentan. - Just on the cryocut. The head of the institute is not satisfied with the quality and wants us to do better. Our usual method is to make a OCT-layer on a chuck, let it freeze, give the tissue on it, surround with OCT, put the cold weight on it, let it freeze and make the sections. We tried putting the tissue in the liquid OCT on the chuck without a layer and freezing it in a mold with the chuck on top. I also tried first freezing the piece solely in liquid nitrogen or isopentan, then putting it on a chuck. All in all - my boss wasn't happy with it. Any input will be appreciated. Thank you in advance Gudrun Lang ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 30 **************************************** From gu.lang <@t> gmx.at Wed Nov 21 12:36:14 2007 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Wed Nov 21 12:36:21 2007 Subject: AW: [Histonet] frozens from lung-tissue In-Reply-To: <014301c82c69$f83573e0$6901a8c0@CHERYLSLAPTOP> Message-ID: <000301c82c6d$65f01770$6412a8c0@dielangs.at> The reason is more the antipathy paired with unfamiliarity with this reagens. I tried it with a small amount of liquid nitrogen in a plasticbox, where I placed the (metal) mold into it, with the tissue and the OCT until it cracks most times. I thought it would be the fastest way of freezing, but the result didn't suit. So I am not very ambitious to do it again. The doctor always speaks about "over-freezing" of the edges and a overall bad morphology. Thank you for the hint with the cryobath. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: Cheryl R. Kerry [mailto:tkngflght@yahoo.com] Gesendet: Mittwoch, 21. November 2007 19:12 An: gu.lang@gmx.at; histonet@lists.utsouthwestern.edu Betreff: RE: [Histonet] frozens from lung-tissue Gudrun--I suspect this isn't quite what you were looking for but have you guys tried a cryobath? It's a little blue countertop 'freezer' unit that has well of pre-chilled isopentane in it, always cold enough to snap freeze without the drama of liquid nitrogen. It is about the size of a big toaster and is ready to go 24/7 with minimal maintenance (temp chart and periodic cleaning). It wasn't overly expensive from what I remember for the great quality of the end results. We mounted the tissue on a pre-frozen OCT-covered chuck, covered it with liquid OCT, lowered it into the cryobath for about 30-40 second and cut...beautiful, artifact free, duplicable and fast. What are the reasons s/he doesn't want the more standard snap freeze methods? Is it a time or expense issue? Cheryl -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Wednesday, November 21, 2007 11:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] frozens from lung-tissue Hi to all, I need the help of those, who have experience on making good frozens from lung-tissue. We look for a method for freezing lung-tissue without the usage of liquid nitrogen, co2 or isopentan. - Just on the cryocut. The head of the institute is not satisfied with the quality and wants us to do better. Our usual method is to make a OCT-layer on a chuck, let it freeze, give the tissue on it, surround with OCT, put the cold weight on it, let it freeze and make the sections. We tried putting the tissue in the liquid OCT on the chuck without a layer and freezing it in a mold with the chuck on top. I also tried first freezing the piece solely in liquid nitrogen or isopentan, then putting it on a chuck. All in all - my boss wasn't happy with it. Any input will be appreciated. Thank you in advance Gudrun Lang From nyilmaz <@t> mersin.edu.tr Wed Nov 21 12:42:55 2007 From: nyilmaz <@t> mersin.edu.tr (nyilmaz@mersin.edu.tr) Date: Wed Nov 21 12:44:14 2007 Subject: [Histonet] Fetal Mouse Brain Processing and... Message-ID: <20071121184255.164173176AE@mail.mersin.edu.tr> Hi histonetters, Are there any suggestions on processing mouse fetus brains D15 and D17 except frozen sectioning? We had a lot of sectioning difficulties and artifacts with these FFPE tissues. Furthermore, any suggestions on preserving frozen tissues for re-sectioning for long periods? Dr. Nejat Yilmaz PhD. Mersin, TURKEY ___________________________________ Mersin Universitesi, http://www.mersin.edu.tr From tkngflght <@t> yahoo.com Wed Nov 21 13:15:37 2007 From: tkngflght <@t> yahoo.com (Cheryl R. Kerry) Date: Wed Nov 21 13:16:13 2007 Subject: A longer suggestion--RE: [Histonet] frozens from lung-tissue In-Reply-To: <000301c82c6d$65f01770$6412a8c0@dielangs.at> Message-ID: <015101c82c72$e6536ca0$6901a8c0@CHERYLSLAPTOP> That helps a bit-- Nitrogen and most other reagents dehydrate as well as freeze if they come in direct contact with the wet tissue. Think 'freeze-dried coffee'. Isopentane is less harsh--which is why it's often the contact chemical being chilled by nitrogen in a double-boiler manner. Dehydration on a small scale is inherent with the method. If you cover your tissue with OCT before freezing, it will protect it from this minor dessication while freezing. With a tissue as 'wet' as lung, you also work against a freezing artifact if you let it freeze too slowly, it will crack if you freeze it too quickly. The forces of the structure of the water expanding when freezing will create significant fissures--both big enough to be seen with the naked eye and microscopic ones--so your pathologist does have a legitimate beef. As with most things in life, too little or too much isn't the best way. Putting the freezing weights on the tissue sometimes mush it and distort tissue relationships. The less mechanical pressure you can put on the tissue, the better. This leads to the next suggestion. There are disposable molds--plastic ones--that come just for freezing OCT embedded tissues. They are in several different sizes. You can buy (or have made) a heat-sink bar that is bored out in the correct size for the embedding mold to fit in. It doesn't have to be a purchased heat sink bar--a small plate of aluminum or any heat-conductive metal will help pull the temp down quickly (think cupcake pan). You put a little alcohol in the bored out bar or on the metal to keep the mold from sticking (like PAM kitchen spray for frozens!) then add the mold (cupcake paper liner) then add a little OCT, your tissue and more OCT with the chuck inverted on top into the wet OCT (like a cream-filled cupcake in the making--the lung is the center). Any time you let OCT freeze then add more, you've created a weak junction that is more likely to fracture when cutting. Whenever possible add more OCT to the prior application before it's completely solid to make a stronger union and prevent chunking off the tissue when you cut. Once it's frozen you pull the plastic mold out of the bar and the mold from the OCT frozen tissue (cupcake paper with the now-done cupcake in it out of the tin and strip off the paper ready, mmmmMMmm, making myself hungry), cut the sloppy bits away, mount on the cryostat head and cut. The heat sink bar would come from the manufacturer of the cryostat--or perhaps your physical plant guys could make one if you showed them a picture (our guys were always game to try a new project). Try Michael Vadeboncoeur from Fisher --he responded earlier about the glove liners and hand cream for frozen hands) for the molds: michael.vadeboncoeur@thermofisher.com Sorry this was so long--you can get samples of the molds and try it for nothing lost but a little time and effort. I hope this helps!! Cheryl -----Original Message----- From: Gudrun Lang [mailto:gu.lang@gmx.at] Sent: Wednesday, November 21, 2007 12:36 PM To: 'Cheryl R. Kerry' Cc: histonet@lists.utsouthwestern.edu Subject: AW: [Histonet] frozens from lung-tissue The reason is more the antipathy paired with unfamiliarity with this reagens. I tried it with a small amount of liquid nitrogen in a plasticbox, where I placed the (metal) mold into it, with the tissue and the OCT until it cracks most times. I thought it would be the fastest way of freezing, but the result didn't suit. So I am not very ambitious to do it again. The doctor always speaks about "over-freezing" of the edges and a overall bad morphology. Thank you for the hint with the cryobath. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: Cheryl R. Kerry [mailto:tkngflght@yahoo.com] Gesendet: Mittwoch, 21. November 2007 19:12 An: gu.lang@gmx.at; histonet@lists.utsouthwestern.edu Betreff: RE: [Histonet] frozens from lung-tissue Gudrun--I suspect this isn't quite what you were looking for but have you guys tried a cryobath? It's a little blue countertop 'freezer' unit that has well of pre-chilled isopentane in it, always cold enough to snap freeze without the drama of liquid nitrogen. It is about the size of a big toaster and is ready to go 24/7 with minimal maintenance (temp chart and periodic cleaning). It wasn't overly expensive from what I remember for the great quality of the end results. We mounted the tissue on a pre-frozen OCT-covered chuck, covered it with liquid OCT, lowered it into the cryobath for about 30-40 second and cut...beautiful, artifact free, duplicable and fast. What are the reasons s/he doesn't want the more standard snap freeze methods? Is it a time or expense issue? Cheryl -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Wednesday, November 21, 2007 11:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] frozens from lung-tissue Hi to all, I need the help of those, who have experience on making good frozens from lung-tissue. We look for a method for freezing lung-tissue without the usage of liquid nitrogen, co2 or isopentan. - Just on the cryocut. The head of the institute is not satisfied with the quality and wants us to do better. Our usual method is to make a OCT-layer on a chuck, let it freeze, give the tissue on it, surround with OCT, put the cold weight on it, let it freeze and make the sections. We tried putting the tissue in the liquid OCT on the chuck without a layer and freezing it in a mold with the chuck on top. I also tried first freezing the piece solely in liquid nitrogen or isopentan, then putting it on a chuck. All in all - my boss wasn't happy with it. Any input will be appreciated. Thank you in advance Gudrun Lang From soofias2 <@t> yahoo.com Wed Nov 21 13:18:10 2007 From: soofias2 <@t> yahoo.com (soofia siddiqui) Date: Wed Nov 21 13:18:12 2007 Subject: AW: [Histonet] frozens from lung-tissue In-Reply-To: <000301c82c6d$65f01770$6412a8c0@dielangs.at> Message-ID: <182180.54616.qm@web39506.mail.mud.yahoo.com> I get the best quality sections when I snap freeze the tissue ( very small not lungs) in dry ice and isopropenal bath. Add a little isopropenal into dry ice to make the bath , use small plastic beem/ beam ( Energy Beam Sciences) capsule, fill with liquid OCT, place the tissue in the capsule with liquid OCT, put hanging tag( some thing to hold into bath or information), cover the capsule and dip the capsule for few seconds. You can see the freezing of OCT, just pull it out from the bath. We in our lab usually leave the frozen tissue in -80 Celsius degrees for over night and then cut next day. Soofia UWHC Madison WI --------------------------------- Never miss a thing. Make Yahoo your homepage. From arunams <@t> interchange.ubc.ca Wed Nov 21 13:23:37 2007 From: arunams <@t> interchange.ubc.ca (Aruna Somasiri) Date: Wed Nov 21 13:23:45 2007 Subject: [Histonet] Help with JB-4 Message-ID: <3287356.4381195673017695.JavaMail.myubc2@brahms.my.ubc.ca> Hi All, I have a problem with my JB-4 blocks and any help is appreciated. The problem is when the block cracks and crumbles when it hits the knife. Any ideas why this is happening and any way to make the block soft? Thanks again Aruna From jengirl1014 <@t> yahoo.com Wed Nov 21 13:25:11 2007 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Wed Nov 21 13:25:20 2007 Subject: [Histonet] Re: Histonet Digest, Vol 48, Issue 30 Message-ID: <324738.14713.qm@web62404.mail.re1.yahoo.com> Happy Thanksgiving everyone and good luck eith the strike tomorrow! Jen ----- Original Message ---- From: "histonet-request@lists.utsouthwestern.edu" To: histonet@lists.utsouthwestern.edu Sent: Wednesday, November 21, 2007 1:01:47 PM Subject: Histonet Digest, Vol 48, Issue 30 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Re: Histogel Question (Dana Settembre) 2. RE: How to keep cryostating without getting frostbite? (Andrea Grantham) 3. RE: Anti-dog IgG (Abigail M. Butler) 4. -80 degree freezer, repair question (Monfils, Paul) 5. frozens from lung-tissue (Gudrun Lang) ---------------------------------------------------------------------- Message: 1 Date: Wed, 21 Nov 2007 11:37:44 -0500 From: "Dana Settembre" Subject: Re: [Histonet] Histogel Question To: , "Abigail M. Butler" Message-ID: Content-Type: text/plain; charset=US-ASCII Abbie, I have used Histogel to embed very small human eye biopsies. What do you mean, "that obviously is not placed in a tube?"? We pick up the small specimen carefully with forceps and place the tissue at the very bottom of the gel and when it goes into a cassette, it is placed facing down. We make sure that the embedding histotech understands exactly where it is for processing and embedding. Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ USA >>> "Abigail M. Butler" 11/21/07 10:13 AM >>> I completely understand how to use histogel to embed free floating cells. But how would I embed a piece of tissue that obviously is not placed in a tube? I am considering purchasing Histogel for the first time and have many ideas of how I could use it, just not sure the proper steps to take to embed let's say two mice eyes. Thanks Abbie Butler, HT (ASCP), QIHC University of Georgia College of Veterinary Medicine _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Wed, 21 Nov 2007 10:22:36 -0700 From: Andrea Grantham Subject: RE: [Histonet] How to keep cryostating without getting frostbite? To: "Monfils, Paul" , Message-ID: <6.2.3.4.1.20071121100954.01faafa8@algranth.inbox.email.arizona.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I cut little rectangular pieces of styrofoam from any of the cartons that are shipped in with cold or frozen items and "glue" them to each side of the knife holder with OCT. This provides a great area to rest my hands while having to manipulate a frozen section. The styrofoam doesn't get cold and it is a comfty place to put your hands and also a slide if you have to pick up multiple sections on one slide. The tissue and OCT doesn't thaw or the slide doesn't get too cold for another section to attach. This excellent piece of advice came from Gayle Callis - thank you again Gayle!!! PS: When I want to clean out the cryostat I just turn it off and take out the knife holder and when the OCT warms up the styrofoam comes off and can be reattached when the cryostat is all clean and ready to be turned back on. I was reading where somebody suggested something to a manufacturer and the idea was not readily accepted and I likewise have made what I thought were excellent suggestions and was met with similar responses - this styrofoam thing was just one of them. Andi At 09:16 AM 11/21/2007, Monfils, Paul wrote: >Just having your hands in the cold air isn't usually the problem. >Each tech, according to their particular style, usually rests their >hand on the knife holder in some manner for stability while picking >up the sections on the slides. The rate of extraction of heat by >contact with -20 degree metal is much greater than the rate by >simple exposure to -20 degree air. It is these points of repeated >contact between skin and metal that usually suffer from >frostbite. I have tried various kinds of gloves and slip-on finger >protectors, but in the final analysis I find that bandaids carefully >applied to the specific points of contact provide enough insulation >to prevent skin damage. I buy the extra wide ones. > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf > of jstaruk > > Sent: Tuesday, November 20, 2007 7:49 PM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] How to keep cryostating without getting frostbite? > > > > Hey all, > > > > Are there any tricks on how to keep your fingers from getting frostbite > > while doing cryostat sectioning for 8 hours a day? My poor techs who have > > been cutting frozen sections for several straight days now are starting to > > complain about these inhumane conditions! Do all of you MOHS techs have > > bleeding, chapped hands and numb fingertips? > > > > Thank you in advance for any suggestions and Happy Thanksgiving to all! > > > > Jim > > > > _____________________ > > Jim Staruk > > Mass Histology Service > > www.masshistology.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 ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html ------------------------------ Message: 3 Date: Wed, 21 Nov 2007 12:28:14 -0500 (EST) From: "Abigail M. Butler" Subject: [Histonet] RE: Anti-dog IgG To: histonet@lists.utsouthwestern.edu Message-ID: <20071121122814.JJF06401@punts1.cc.uga.edu> Content-Type: text/plain; charset=us-ascii MaryAnn, I use only anti-dog IgG.....and my dilution is 1:150,000. Crazy, isn't it? I also used HIER for pretreatment with citrate buffer. There is not much background. Hope this helps Abbie Butler, HT (ASCP), QIHC University of Georgia College of Veterinary Medicine ------------------------------ Message: 4 Date: Wed, 21 Nov 2007 12:36:05 -0500 From: "Monfils, Paul" Subject: [Histonet] -80 degree freezer, repair question To: Message-ID: <4EBFF65383B74D49995298C4976D1D5E273CFC@LSRIEXCH1.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" We have a rather old Baxter Scientific Cryo-Fridge chest type -80 degree freezer that works fine except the lid gasket is shot, so cold air leaks out and frost and ice build up around the edges of the lid. Does anyone know of someone who could replace this gasket? Or somewhere we could purchase the part to install ourselves? We are also unable to find particle filters for the front door of the compressor compartment. ------------------------------ Message: 5 Date: Wed, 21 Nov 2007 18:50:42 +0100 From: "Gudrun Lang" Subject: [Histonet] frozens from lung-tissue To: Message-ID: <000601c82c67$09d1b0d0$6412a8c0@dielangs.at> Content-Type: text/plain; charset="us-ascii" Hi to all, I need the help of those, who have experience on making good frozens from lung-tissue. We look for a method for freezing lung-tissue without the usage of liquid nitrogen, co2 or isopentan. - Just on the cryocut. The head of the institute is not satisfied with the quality and wants us to do better. Our usual method is to make a OCT-layer on a chuck, let it freeze, give the tissue on it, surround with OCT, put the cold weight on it, let it freeze and make the sections. We tried putting the tissue in the liquid OCT on the chuck without a layer and freezing it in a mold with the chuck on top. I also tried first freezing the piece solely in liquid nitrogen or isopentan, then putting it on a chuck. All in all - my boss wasn't happy with it. Any input will be appreciated. Thank you in advance Gudrun Lang ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 30 **************************************** ____________________________________________________________________________________ Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. http://mobile.yahoo.com/sports;_ylt=At9_qDKvtAbMuh1G1SQtBI7ntAcJ From vsnider <@t> shrinenet.org Wed Nov 21 13:39:21 2007 From: vsnider <@t> shrinenet.org (Snider, Vivian Deanna) Date: Wed Nov 21 13:39:29 2007 Subject: [Histonet] (no subject) Message-ID: <84BE46B37B314D409C5A17B7BAB022D601A083E1@IDC-EX-VS01.shriners.cc> Jim, There are days my hands are in the cryostat the entire day. I have problems with my pinkie finger getting so cold it hurts. What I have done to offset this is, first I cut one finger out of a cotton glove we use for getting into the walkin and I put that on my pinkie. Then I put on a nylon liner, (from Fisher) and then the nitrile glove. It does seem to help. I can get through 3 to 4 hours before I have to take a break. Hope it helps as silly as it sounds! Deanna Snider HT ASCP Lead Histotechnician Shriners Hospital for Children Cincinnati, Oh Message: 9 Date: Tue, 20 Nov 2007 19:49:17 -0500 From: "jstaruk" Subject: [Histonet] How to keep cryostating without getting frostbite? To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hey all, Are there any tricks on how to keep your fingers from getting frostbite while doing cryostat sectioning for 8 hours a day? My poor techs who have been cutting frozen sections for several straight days now are starting to complain about these inhumane conditions! Do all of you MOHS techs have bleeding, chapped hands and numb fingertips? Thank you in advance for any suggestions and Happy Thanksgiving to all! Jim 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 CIngles <@t> uwhealth.org Wed Nov 21 13:42:02 2007 From: CIngles <@t> uwhealth.org (Ingles Claire) Date: Wed Nov 21 13:42:07 2007 Subject: [Histonet] How to keep cryostating without getting frostbite? References: <4EBFF65383B74D49995298C4976D1D5E273CFA@LSRIEXCH1.lsmaster.lifespan.org> <6.2.3.4.1.20071121100954.01faafa8@algranth.inbox.email.arizona.edu> Message-ID: <08A0A863637F1349BBFD83A96B27A50A1200A7@uwhis-xchng3.uwhis.hosp.wisc.edu> Jim: I work in a Mohs lab. I have been working in my cryostat for 3 years and haven't had a problem. We are able to us 'surgical' type gloves. They are thicker and contain your body heat much better. I have sometimes been working in the cryostat with just blue nitriles and definitely notice the difference. We also wear barrier gowns so maybe the extra heat retention helps our hands stay warmer. I also try to touch the metal areas as little as possible. I use forceps (room temp) for tissue manipulation. The thin gloves you are enquiring about actually have no finger tips, so I don't know how much they would help you. It is also important to keep a bottle of lotion around. Specially as winter is setting in. Keeping the hands moisturized helps protect your hands from the cold too. If you use both hands for slide stability during section pick up, you can rest your elbows on the outside frame of the cryostat and not need to touch any of the metal. Hope this helps, Claire ________________________________ At 09:16 AM 11/21/2007, Monfils, Paul wrote: >Just having your hands in the cold air isn't usually the problem. >Each tech, according to their particular style, usually rests their >hand on the knife holder in some manner for stability while picking >up the sections on the slides. The rate of extraction of heat by >contact with -20 degree metal is much greater than the rate by >simple exposure to -20 degree air. It is these points of repeated >contact between skin and metal that usually suffer from >frostbite. I have tried various kinds of gloves and slip-on finger >protectors, but in the final analysis I find that bandaids carefully >applied to the specific points of contact provide enough insulation >to prevent skin damage. I buy the extra wide ones. > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf > of jstaruk > > Sent: Tuesday, November 20, 2007 7:49 PM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] How to keep cryostating without getting frostbite? > > > > Hey all, > > > > Are there any tricks on how to keep your fingers from getting frostbite > > while doing cryostat sectioning for 8 hours a day? My poor techs who have > > been cutting frozen sections for several straight days now are starting to > > complain about these inhumane conditions! Do all of you MOHS techs have > > bleeding, chapped hands and numb fingertips? > > > > Thank you in advance for any suggestions and Happy Thanksgiving to all! > > > > Jim > > > > _____________________ > > Jim Staruk > > Mass Histology Service > > www.masshistology.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 ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Lchausse <@t> nmh.org Wed Nov 21 14:58:47 2007 From: Lchausse <@t> nmh.org (Chaussey, Leslie) Date: Wed Nov 21 14:59:06 2007 Subject: [Histonet] frozens from lung-tissue Message-ID: If you're thinking of the Histobath from ThermoFisher, we were recently told those units were discontinued. ----- Original Message ----- From: histonet-bounces@lists.utsouthwestern.edu To: gu.lang@gmx.at ; histonet@lists.utsouthwestern.edu Sent: Wed Nov 21 12:11:41 2007 Subject: RE: [Histonet] frozens from lung-tissue Gudrun--I suspect this isn't quite what you were looking for but have you guys tried a cryobath? It's a little blue countertop 'freezer' unit that has well of pre-chilled isopentane in it, always cold enough to snap freeze without the drama of liquid nitrogen. It is about the size of a big toaster and is ready to go 24/7 with minimal maintenance (temp chart and periodic cleaning). It wasn't overly expensive from what I remember for the great quality of the end results. We mounted the tissue on a pre-frozen OCT-covered chuck, covered it with liquid OCT, lowered it into the cryobath for about 30-40 second and cut...beautiful, artifact free, duplicable and fast. What are the reasons s/he doesn't want the more standard snap freeze methods? Is it a time or expense issue? Cheryl -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Wednesday, November 21, 2007 11:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] frozens from lung-tissue Hi to all, I need the help of those, who have experience on making good frozens from lung-tissue. We look for a method for freezing lung-tissue without the usage of liquid nitrogen, co2 or isopentan. - Just on the cryocut. The head of the institute is not satisfied with the quality and wants us to do better. Our usual method is to make a OCT-layer on a chuck, let it freeze, give the tissue on it, surround with OCT, put the cold weight on it, let it freeze and make the sections. We tried putting the tissue in the liquid OCT on the chuck without a layer and freezing it in a mold with the chuck on top. I also tried first freezing the piece solely in liquid nitrogen or isopentan, then putting it on a chuck. All in all - my boss wasn't happy with it. Any input will be appreciated. Thank you in advance Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------- This message and any included attachments are intended only for the addressee. The information contained in this message is confidential and may constitute proprietary or non-public information under international, federal, or state laws. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail. From Kathy.Bucknell <@t> leica-microsystems.com Wed Nov 21 15:03:29 2007 From: Kathy.Bucknell <@t> leica-microsystems.com (Kathy.Bucknell@leica-microsystems.com) Date: Wed Nov 21 15:03:39 2007 Subject: [Histonet] Bucknell, Kathy is out of the office. Message-ID: I will be out of the office starting 11/21/2007 and will not return until 11/22/2007. I am at the Northbrook warehouse today and will have limited access to my emails. For immediate assistance, please contact me at 847-224-6766. ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From Rcartun <@t> harthosp.org Wed Nov 21 15:37:10 2007 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Wed Nov 21 15:37:26 2007 Subject: [Histonet] IHC for INI Message-ID: <47445EB60200007700009423@gwmail6.harthosp.org> Is anyone offering immunohistochemical staining for INI in AT/RT brain tumors? 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 From Kathy.Abels <@t> sial.com Wed Nov 21 16:03:29 2007 From: Kathy.Abels <@t> sial.com (Kathy Abels) Date: Wed Nov 21 16:03:54 2007 Subject: [Histonet] Kathy Abels/ops/diag/sial is out of the office. Message-ID: I will be out of the office starting 11/20/2007 and will not return until 11/26/2007. I will respond to your message when I return. For urgent matters, Please contact Leigh Gaskill. This message and any files transmitted with it are the property of Sigma-Aldrich Corporation, are confidential, and are intended solely for the use of the person or entity to whom this e-mail is addressed. If you are not one of the named recipient(s) or otherwise have reason to believe that you have received this message in error, please contact the sender and delete this message immediately from your computer. Any other use, retention, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. From Rcartun <@t> harthosp.org Wed Nov 21 16:25:50 2007 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Wed Nov 21 16:26:07 2007 Subject: [Histonet] re:B-5 solution In-Reply-To: References: Message-ID: <47446A1E020000770000942B@gwmail6.harthosp.org> No one should be using B5. If your pathologist won't take "No" for an answer, get your safety officer involved. We stopped using B5 many years ago and now use only formalin. 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 >>> 11/19/07 2:07 PM >>> We have 1 Pathologist who insists on using B-5 solution for nuclear detail on small biopsies. How many labs are still using B 5 or what is you suggestion other than Zinc Formalin for fixation. Thanks. ************************************** See what's new at http://www.aol.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From michael.owen <@t> fda.hhs.gov Wed Nov 21 18:34:06 2007 From: michael.owen <@t> fda.hhs.gov (Owen, Michael P) Date: Wed Nov 21 18:34:29 2007 Subject: [Histonet] Histology Openings in Seattle (Everett), WA Message-ID: <449E51C6DA0AD840B44F57C7A6EB07BF0160A6F4@FMD3VS022.fda.gov> craigslist seattle-tacoma science/biotech jobs http://seattle.craigslist.org/sno/sci/485851194.html Histology Technician/Histotechnologist Reply to: hr@snblusa.com Date: 2007-11-21, 1:15PM PST SNBL USA, Ltd. Histology Technician/Histotechnologist SNBL USA is a leading contract research organization located just 30 minutes from Seattle, in Everett, WA. Through a commitment to investment and excellence, our team is able to offer the biotechnology and pharmaceutical industries high quality science and service. We are recruiting for a histology technician/histotechnologist. This position will process, section, stain, and prepare slides of tissue samples for microscopic evaluation by pathologists. Responsibilities will include: * Process, embed, section, and perform necessary stains to highlight specific tissue patterns and structures. * Perform quality control (QC) on finished slide preparations. * Maintain required documentation of all procedures and laboratory equipment. * Review related documentation for deviations in Standard Operating Procedures (SOP). * Maintain and troubleshoot equipment operation failures; take appropriate actions to keep interruptions in work flow to a minimum. * Train and oversee histology laboratory personnel/assistants. * Execute special histological and histochemical stains used to emphasize specific tissue elements. * Perform special histological procedures such as frozen sectioning and electron microscopy preparations (e.g. plastic processing, embedding, sectioning), and photomicrography. * May delegate some technical duties to subordinates/laboratory assistants. Education & Experience: * Associate's (A.A.) degree or higher, specializing in anatomy-related medical science. * HT/HTL certification by American Society of Clinical Pathologist (ASCP). * 1+ years related experience and/or training. * Equivalent combination of education/experience will be considered. SNBL USA, Ltd. is a smoke-free, drug-free workplace. Pre-employment drug test required. For more information about SNBL USA, Ltd., please visit our website at www.snblusa.com. For consideration send cover letter and resume to: hr@snblusa.com Equal Opportunity Employer Location: S. Everett Compensation: SNBL USA, Ltd. offers a competitive compensation and benefits package including health, dental, vision, pharmaceutical, life, and disability insurances, 401(k), Section 125, and a generous PTO package. Principals only. Recruiters, please don't contact this job poster. Please, no phone calls about this job! Please do not contact job poster about other services, products or commercial interests. PostingID: 485851194 craigslist seattle-tacoma science/biotech jobs http://seattle.craigslist.org/sno/sci/485856296.html PATHOLOGY ASSOCIATE Reply to: hr@snblusa.com Date: 2007-11-21, 1:21PM PST SNBL USA, Ltd. PATHOLOGY ASSOCIATE SNBL USA is a leading contract research organization located just 30 minutes from Seattle, in Everett, WA. Through a commitment to investment and excellence, our team is able to offer the biotechnology and pharmaceutical industries high quality science and service. We are recruiting for a Pathology Associate. The primary role of the Pathology Associate will be to perform all gross pathology related responsibilities including tissue trimming and processing of all pathology related study subjects as specified in company protocols and in compliance with GLP standards. RESPONSIBILITIES * Perform gross pathology procedures * Perform histology trimming procedures * Inventory tissue blocks * Perform room preparation and equipment maintenance * Perform instrument maintenance * Maintain stock and working solutions for mandatory fixatives and other chemicals * Recording pertinent data, written or computer input, for necropsy, trimming, and maintenance. EDUCATION and/or EXPERIENCE * High school diploma or equivalent, BS degree in a science discipline preferred * 2+ years of biological science laboratory work * Excellent knowledge in anatomical biology * Basic chemistry required * Aptitude to learn new histology techniques in pathology services * Ability to read, interpret, and implement client protocol procedures SNBL USA, Ltd. is a smoke-free, drug-free workplace. Pre-employment drug test required. For more information about SNBL USA, Ltd., please visit our website at www.snblusa.com. For consideration send cover letter and resume to: hr@snblusa.com Equal Opportunity Employer Location: S. Everett Compensation: SNBL USA, Ltd. offers a competitive compensation and benefits package including health, dental, vision, pharmaceutical, life, and disability insurances, 401(k), Section 125, and a generous PTO package. Principals only. Recruiters, please don't contact this job poster. Please, no phone calls about this job! Please do not contact job poster about other services, products or commercial interests. PostingID: 485856296 craigslist seattle-tacoma science/biotech jobs http://seattle.craigslist.org/sno/sci/485854210.html HISTOLOGY LABORATORY MANAGER Reply to: hr@snblusa.com Date: 2007-11-21, 1:19PM PST SNBL USA, Ltd. HISTOLOGY LABORATORY MANAGER SNBL USA is a leading contract research organization located just 30 minutes from Seattle, in Everett, WA. Through a commitment to investment and excellence, our team is able to offer the biotechnology and pharmaceutical industries high quality science and service. We are recruiting for a Histology Laboratory Manager. The primary role of the Histology Laboratory Manager will be to direct and manage the daily operations and supervise the staff of the histopathology department to ensure the highest quality product for histopathology evaluation by pathologists, study directors, and clients according to Standard Operating Procedures (SOP) and regulations. RESPONSIBILITIES * Initiate, acquire and oversee installation and operation of all equipment and instruments necessary to adequately perform histology, immunohistochemistry, plastic, electron micrography, photomicrography, and other related services. * Develop and implement laboratory policies, departmental Standard Operating Procedures (SOPs) and validation/qualification requirements, ensuring compliance with FDA (GLP), L&I, and other regulatory agencies for study validity. * Delegate and supervise appropriate responsibilities for histology laboratory personnel. * Identify and troubleshoot problems with personnel, equipment and procedures. * Coordinate and perform necropsy work, trimming, processing, microtomy, staining and other technical duties as required. * Control histology work schedule to meet study timelines. * Supervise all administrative responsibilities as related to staffing including analyzing future staffing needs. * Conduct annual staff evaluations and enforces safety regulations. * Maintain inventory control in cooperation with Laboratory Support Services. * Evaluate capital purchases and contracts with appropriate vendors, writes purchasing proposals and constructs budget requirements. * Create and implement training programs to establish and maintain quality specimens using uniform methods in procedures and current techniques. EDUCATION and/or EXPERIENCE * Bachelor of Science degree specializing in histology or related science and HT or HTL (ASCP) certification or equivalent. * Must have eight years recent experience and/or training. * 2+ years of demonstrated managerial experience in a laboratory setting. SNBL USA, Ltd. is a smoke-free, drug-free workplace. Pre-employment drug test required. For more information about SNBL USA, Ltd., please visit our website at www.snblusa.com. For consideration send cover letter and resume to: hr@snblusa.com Equal Opportunity Employer Location: S. Everett Compensation: SNBL USA, Ltd. offers a competitive compensation and benefits package including health, dental, vision, pharmaceutical, life, and disability insurances, 401(k), Section 125, and a generous PTO package. Principals only. Recruiters, please don't contact this job poster. Please, no phone calls about this job! Please do not contact job poster about other services, products or commercial interests. PostingID: 485854210 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.hhs.gov From mickie25 <@t> netzero.net Wed Nov 21 19:20:33 2007 From: mickie25 <@t> netzero.net (Mickie Johnson) Date: Wed Nov 21 19:21:20 2007 Subject: [Histonet] -80 degree freezer, repair question In-Reply-To: <4EBFF65383B74D49995298C4976D1D5E273CFC@LSRIEXCH1.lsmaster.lifespan.org> References: <4EBFF65383B74D49995298C4976D1D5E273CFC@LSRIEXCH1.lsmaster.lifespan.org> Message-ID: Paul, You can purchase different types of refrigerator gasket material from an appliance parts supply store. The cross-section shape comes in 'C', 'G', 'M' and many types. You will need to know what your looks like. Maybe take a thin slice of yours to compare with what they have available? Good Luck! Mickie Mickie Johnson, B.S., HTL(ASCP) Mohs Histology Consulting Services, LLC & Mohs Lab Staffing 2507 S. Manito Blvd. Spokane, WA 99203 509-954-7134 Web: www.mohshistotemp.com & www.mohslabstaffing.com Email: mickie25@netzero.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Wednesday, November 21, 2007 9:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] -80 degree freezer, repair question We have a rather old Baxter Scientific Cryo-Fridge chest type -80 degree freezer that works fine except the lid gasket is shot, so cold air leaks out and frost and ice build up around the edges of the lid. Does anyone know of someone who could replace this gasket? Or somewhere we could purchase the part to install ourselves? We are also unable to find particle filters for the front door of the compressor compartment. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From aakrasht <@t> yahoo.com Wed Nov 21 20:23:38 2007 From: aakrasht <@t> yahoo.com (Ali A. Krasht) Date: Wed Nov 21 20:23:43 2007 Subject: [Histonet] HELP Need a Sample Business Plan in Histology Message-ID: <643727.15551.qm@web50909.mail.re2.yahoo.com> Hi There Anyone would like to help, I need a sample business plan for opening a Histology lab (human tissue in Dallas Texas); basically focusing on technical and surgical dermatological samples where its going to involve all aspects in Histology from mohs,special stains,immunos,mycology,embedding, cutting, staining etc... so I can have an idea in how to plan or write, specially the projection part is given me trouble. I got all the outline from the Small Business Bureau on how to write it but I need a little help by sending me a sample plan please, I could not find any online. Thanks for your time and attention on this matter. Ali ____________________________________________________________________________________ Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. http://overview.mail.yahoo.com/ From Kemlo.Rogerson <@t> waht.swest.nhs.uk Thu Nov 22 03:41:50 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu Nov 22 03:41:56 2007 Subject: [Histonet] frozens from lung-tissue Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F204@wahtntex2.waht.swest.nhs.uk> You can't practically get any colder than liquid nitrogen and if you don't want ice artefact then I guess you have to use it. The only issue sometimes is that you can get a bubble of nitrogen around the tissue that acts as an insulator; maybe you can freeze in/ on metal foil? What are the problems you have or why is your boss unhappy? If it's ice crystal artefact you must freeze faster, if it's nuclear staining, try post fixing the slides before staining in an alcohol. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From peter.craven <@t> haht.scot.nhs.uk Thu Nov 22 08:14:48 2007 From: peter.craven <@t> haht.scot.nhs.uk (PETER CRAVEN) Date: Thu Nov 22 08:17:01 2007 Subject: [Histonet] Frozens from Lung tissue Message-ID: <70B1BBF61EF50C488725E84760EC7BFF0458186D@NHSHRMDC02.NHSH.SCOT.NHS.UK> Gudrun We used to embed our lung blocks in Gelatin prior to freezing in OCT to help hold the lung together when frozen sectioned Rinse out the formalin 10% Gelatin till the block of tissue sinks 20% Gelatin overnight Change to fresh 20% and set in a fridge to set the gelatin Trim off the excess gelatin Adhere to cryostat block with OCT and cut Hope this helps Peter Peter L Craven FIBMS BMS 2 Pathology Department Raigmore Hospital Inverness IV2 3UJ tel. 01463 704000 ext. 4269 email: peter.craven@haht.scot.nhs.uk ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the system manager. This footnote also confirms that this email message has been swept by MIMEsweeper for the presence of computer viruses. www.mimesweeper.com ********************************************************************** Please consider the environment before printing this e-mail From jstaruk <@t> masshistology.com Thu Nov 22 10:13:16 2007 From: jstaruk <@t> masshistology.com (jstaruk) Date: Thu Nov 22 10:14:09 2007 Subject: [Histonet] HELP Need a Sample Business Plan in Histology In-Reply-To: <643727.15551.qm@web50909.mail.re2.yahoo.com> Message-ID: <1F2DD84F46E247F1A6C3704AB705A3B3@JimPC> Try this page: http://www.download.com/3120-20_4-0.html?tg=dl-20&qt=business%20plan&tag=src h Jim _____________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ali A. Krasht Sent: Wednesday, November 21, 2007 9:24 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HELP Need a Sample Business Plan in Histology Hi There Anyone would like to help, I need a sample business plan for opening a Histology lab (human tissue in Dallas Texas); basically focusing on technical and surgical dermatological samples where its going to involve all aspects in Histology from mohs,special stains,immunos,mycology,embedding, cutting, staining etc... so I can have an idea in how to plan or write, specially the projection part is given me trouble. I got all the outline from the Small Business Bureau on how to write it but I need a little help by sending me a sample plan please, I could not find any online. Thanks for your time and attention on this matter. Ali ____________________________________________________________________________ ________ Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. http://overview.mail.yahoo.com/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gagnone <@t> KGH.KARI.NET Thu Nov 22 11:01:31 2007 From: gagnone <@t> KGH.KARI.NET (Gagnon, Eric) Date: Thu Nov 22 11:01:37 2007 Subject: [Histonet] TAZ protein and uses Message-ID: Has anyone tried immunohistochemistry using the TAZ-protein antibody? (alternate names BTHS, CMD3A, EFE) A researcher has asked us to try it on the Ventana Benchmark XT, even though the product insert sheet lists Western Blot and ELISA as uses. Perhaps the larger question is: should IHC be attempted using antibodies that do not list FFPE tissue as one of the antibody's uses? Eric Gagnon MLT Histology Laboratory Kingston General Hospital Kingston, Ontario, Canada From RSRICHMOND <@t> aol.com Thu Nov 22 19:47:17 2007 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Thu Nov 22 19:47:22 2007 Subject: [Histonet] Re: B5 Message-ID: Richard Cartun notes: >>No one should be using B5. If your pathologist won't take "No" for an answer, get your safety officer involved. We stopped using B5 many years ago and now use only formalin.<< I agree completely. This is a dead issue. You can't use B5, or other fixatives containing mercury such as Zenker's fixative. I don't think any present fixative offers significant advantages over neutral buffered formalin. You have to prepare the specimen properly (thin sections of lymph node) and fix overnight. - I, too, lament the passing of mercury and chromium fixatives. Richard Cartun - I was in residency with a pathologist named Martin Berman who was on the staff of Hartford Hospital for quite a while. He seems to have dropped off the face of the eartth. Do you know him? Bob Richmond Samurai Pathologist Knoxville TN From gu.lang <@t> gmx.at Fri Nov 23 09:14:45 2007 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Nov 23 09:14:57 2007 Subject: AW: [Histonet] frozens from lung-tissue In-Reply-To: <86ADE4EB583CE64799A9924684A0FBBF0222F204@wahtntex2.waht.swest.nhs.uk> Message-ID: <000001c82de3$96ddbf00$6412a8c0@dielangs.at> Kemlo, My original question was, how to get the best frozen lung-slides without the use of liquid nitrogen or isopentan. - just with the cryocut, chucks and OCT. The pathologist complains about "overfrozen" edges and a bad overall morphology. I will ask him for more descriptionary details or get a picture. Thank you for your hints. Gudrun Lang Biomed. Analytikerin Histolabor Akh Linz Krankenhausstr. 9 4020 Linz +43(0)732/7806-6754 -----Urspr?ngliche Nachricht----- Von: Kemlo Rogerson [mailto:Kemlo.Rogerson@waht.swest.nhs.uk] Gesendet: Donnerstag, 22. November 2007 10:42 An: gu.lang@gmx.at; histonet@lists.utsouthwestern.edu Betreff: RE: [Histonet] frozens from lung-tissue You can't practically get any colder than liquid nitrogen and if you don't want ice artefact then I guess you have to use it. The only issue sometimes is that you can get a bubble of nitrogen around the tissue that acts as an insulator; maybe you can freeze in/ on metal foil? What are the problems you have or why is your boss unhappy? If it's ice crystal artefact you must freeze faster, if it's nuclear staining, try post fixing the slides before staining in an alcohol. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From Kemlo.Rogerson <@t> waht.swest.nhs.uk Fri Nov 23 09:23:49 2007 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Fri Nov 23 09:23:55 2007 Subject: [Histonet] frozens from lung-tissue Message-ID: <86ADE4EB583CE64799A9924684A0FBBF0222F24C@wahtntex2.waht.swest.nhs.uk> Kemlo, My original question was, how to get the best frozen lung-slides without the use of liquid nitrogen or isopentan. - just with the cryocut, chucks and OCT. The pathologist complains about "overfrozen" edges and a bad overall morphology. I will ask him for more descriptionary details or get a picture. Thank you for your hints. Gudrun Lang But that's the thing I'm struggling with. Liquid nitrogen with or without 'additives' is the best way of reducing ice artefact, so by not using it I expect you to get worse results. If you want 'better' morphology then post fix. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From gu.lang <@t> gmx.at Fri Nov 23 09:56:10 2007 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Nov 23 09:56:18 2007 Subject: AW: [Histonet] frozens from lung-tissue In-Reply-To: <86ADE4EB583CE64799A9924684A0FBBF0222F24C@wahtntex2.waht.swest.nhs.uk> Message-ID: <000101c82de9$5df5d140$6412a8c0@dielangs.at> Sure, you are right. I hoped to go a step up on the ladder of quality with some small changes in our method, because we are not familiar with the real snap-freezing. For all other tissues the simple method is good enough, just the lung makes a headache. And this special situation only occurs once in two months and only with our primarius (head of institute). Gudrun Lang -----Urspr?ngliche Nachricht----- Von: Kemlo Rogerson [mailto:Kemlo.Rogerson@waht.swest.nhs.uk] Gesendet: Freitag, 23. November 2007 16:24 An: gu.lang@gmx.at Cc: histonet@lists.utsouthwestern.edu Betreff: RE: [Histonet] frozens from lung-tissue Kemlo, My original question was, how to get the best frozen lung-slides without the use of liquid nitrogen or isopentan. - just with the cryocut, chucks and OCT. The pathologist complains about "overfrozen" edges and a bad overall morphology. I will ask him for more descriptionary details or get a picture. Thank you for your hints. Gudrun Lang But that's the thing I'm struggling with. Liquid nitrogen with or without 'additives' is the best way of reducing ice artefact, so by not using it I expect you to get worse results. If you want 'better' morphology then post fix. Kemlo Rogerson Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; The road to success is dotted with many tempting parking places. --Author Unknown This e-mail is confidential and privileged. If you are not the intended recipient please accept my 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 me that this message has gone astray before deleting it. Thank you for your co-operation From pruegg <@t> ihctech.net Fri Nov 23 12:52:21 2007 From: pruegg <@t> ihctech.net (patsy ruegg) Date: Fri Nov 23 12:47:30 2007 Subject: [Histonet] TAZ protein and uses In-Reply-To: Message-ID: <200711231847.lANIlHgj003983@pro12.abac.com> Yes,. I do ihc all the time on antibodies listed for use on westerns and elisa only. Sometimes they work on ffpe tissue and sometimes they don't. just establish testing criteria and do use the dilution recommendations for elisa/westerns as a guide, in general I usually concentrate 10x for ihc over elisa recommendations but you still have to do a titer range comparing several dilutions and times, also you will need to try several AR pretreatments, generally I use HIER with lph and hph buffer, I also try some enzyme digestion proteinase k, pepsin, etc. until I get something that works, the key is to have good positive control tissue to test. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gagnon, Eric Sent: Thursday, November 22, 2007 10:02 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] TAZ protein and uses Has anyone tried immunohistochemistry using the TAZ-protein antibody? (alternate names BTHS, CMD3A, EFE) A researcher has asked us to try it on the Ventana Benchmark XT, even though the product insert sheet lists Western Blot and ELISA as uses. Perhaps the larger question is: should IHC be attempted using antibodies that do not list FFPE tissue as one of the antibody's uses? Eric Gagnon MLT Histology Laboratory Kingston General Hospital Kingston, Ontario, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Fri Nov 23 14:05:58 2007 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Nov 23 14:06:15 2007 Subject: [Histonet] 35 mm transfer device Message-ID: <4746EC5602000077000094BF@gwmail6.harthosp.org> Does anyone have a recommendation for a device that will transfer "old" 35 mm color slides into "modern day" digital images? 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 From jstaruk <@t> masshistology.com Fri Nov 23 14:17:19 2007 From: jstaruk <@t> masshistology.com (jstaruk) Date: Fri Nov 23 14:18:11 2007 Subject: [Histonet] 35 mm transfer device In-Reply-To: <4746EC5602000077000094BF@gwmail6.harthosp.org> Message-ID: <81687705AA1042639F5B1FD4079E6B65@JimPC> My Epson Stylus CX7800 printer / scanner came with a neat little device that scans (I think) 6 at a time and saves them as individual digital images. I transferred all of my Kodachromes to digital this way. They look great! Jim _____________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Friday, November 23, 2007 3:06 PM To: Histonet Subject: [Histonet] 35 mm transfer device Does anyone have a recommendation for a device that will transfer "old" 35 mm color slides into "modern day" digital images? 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gayle.callis <@t> bresnan.net Fri Nov 23 14:31:20 2007 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Fri Nov 23 14:30:18 2007 Subject: [Histonet] Attn: Richard Cartun 35 mm slide scanner Message-ID: <001401c82e0f$ce824490$6501a8c0@DHXTS541> Richard, I ran across this one in the last week from a membership discount store, but you should be able to buy it elsewhere. PrimeFilm 35mm film and slide scanner 365OU, Digital Ict technology, and includes Adobephotoshop Elements 4.0. It had an list price of $249.00. If you Google this device, it probably can be purchased for as little as $150, as it was from COSTCO. I was impressed by the simplicity of the device - as seen in photograph in advertisement. This device had a flat design and didn't look like a desktop hog. We have a Nikon film and slide scanner, but it is pricey, well over $500. Good luck finding this - Happy Holidays Gayle Callis HT,HTL,MT(ASCP) From gayle.callis <@t> bresnan.net Fri Nov 23 14:48:59 2007 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Fri Nov 23 14:48:01 2007 Subject: [Histonet] Cryotomy and preventing frost bite Message-ID: <001b01c82e12$462be530$6501a8c0@DHXTS541> We use the styrofoam square method mentioned by Andi Grantham, but you can also use the little cork self stick pads used to protect inside corners of cupboard doors. Any hardware store has these in different sizes. It is best to stick these on when the knife holder it warm, and clean plus you can build these up to a preferred height. The styrofoam square is certainly cheap, and we simply use Scotch self stick tape on back so it can be moved to a place comfortable per operator. Leica Microsystems had addressed this problem with a nice hand/finger rest, removable if not desired, on their new 1950(?) cryostat knifeholder. It was quite comfortable, and I now hope they will retrofit their 1850 knife holder with a similar device for the brush method cryotomy afficianados (sp?) To protect hands from cold and dishpan hand "perspiration", we also buy silk glove liners from a company called Winter Silks. These are thin, right/left handed, come in black or white, different sizes, and seconds are cheaper. There is no loss of dexterity when you put a latex or nitrile glove over these. We wash them with hand soap after using. Gloves that are stretchy or elasticized tend to cut the circulation in hands and fingers, and put pressure on painful joints, but the silk gloves do not have any spandex in them. Winter Silks has a website. Gayle Callis HT,HTL,MT(ASCP) Bozeman MT From gayle.callis <@t> bresnan.net Fri Nov 23 14:50:46 2007 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Fri Nov 23 14:49:44 2007 Subject: [Histonet] Correction of spelling for 35 mm slide scanner name Message-ID: <002401c82e12$85ad1120$6501a8c0@DHXTS541> Digital Ice, not Ict! Gayle Callis From gayle.callis <@t> bresnan.net Fri Nov 23 15:10:25 2007 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Fri Nov 23 15:09:26 2007 Subject: [Histonet] GMA block sectioning problems Message-ID: <001601c82e15$4684d340$6501a8c0@DHXTS541> You did not say what type of knife you are using? Nor the section thickness you are working with/desire? You can try softening the plastic by putting it into a moist chamber for a short time, but you should NOT touch the block if it is wet unless you wear nitrile gloves. Too much water on a block face usually means a section that is mush as it comes off onto blade. When trimming the block, do not try to trim it as you would a paraffin block, the thicker the micrometer setting, the more likely you will have crumbled sections. Approach the block face with care, and not taking thick "trim" - a sharp blade/knife it critical for good trimming too. GMA was developed for getting THIN sections, 3 um down to 1 um. You can section at 4 or 5um successfull, but the thicker you go may mean crumbled sections. We always sectioned with glass triangular knives, but people do have success with disposable blade, they must be VERY sharp, and probably a brand new edge after trimming or you can use tungsten carbide knives. There are triangular tungsten carbide knives available from Delaware Diamond Knives (DDK). Finally, you may need to adjust the plasticizer content, i.e. increase the amount a bit in order soften the plastic or make it a bit more flexible,. The spec sheets that come with the kit should tell you how to do this, OR you can contact Poly Sciences tech services and ask them how to do this. Gayle Callis HT,HTL,MT(ASCP) Bozeman MT From llewllew <@t> shaw.ca Fri Nov 23 16:53:30 2007 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Nov 23 16:54:37 2007 Subject: [Histonet] 35 mm transfer device References: <4746EC5602000077000094BF@gwmail6.harthosp.org> Message-ID: <000801c82e23$ab363a50$05024246@yourlk4rlmsu> I use an Epson 4870 scanner. It can scan at 4,000 dpi, which is good enough for 35 mm negatives and transparencies. The negatives can then be reversed to positives. It comes with masks for several film formats, including 2x2 and 35 mm strips. The model number may have changed since I bought mine. Bryan Llewellyn ----- Original Message ----- From: "Richard Cartun" To: "Histonet" Sent: Friday, November 23, 2007 12:05 PM Subject: [Histonet] 35 mm transfer device Does anyone have a recommendation for a device that will transfer "old" 35 mm color slides into "modern day" digital images? 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From beingmary53 <@t> sbcglobal.net Fri Nov 23 17:14:26 2007 From: beingmary53 <@t> sbcglobal.net (MARY JOHNSON) Date: Fri Nov 23 17:14:32 2007 Subject: [Histonet] heatoxylin Message-ID: <850398.25222.qm@web81611.mail.mud.yahoo.com> we use harris hematoxylin and to every 400ml we added 15ml of glacial acetic acid. hope this help. From aakrasht <@t> yahoo.com Fri Nov 23 19:16:44 2007 From: aakrasht <@t> yahoo.com (Ali A. Krasht) Date: Fri Nov 23 19:16:52 2007 Subject: [Histonet] Need a Sample Business Plan in Histology HELP Message-ID: <898405.69563.qm@web50909.mail.re2.yahoo.com> Hi There Anyone would like to help, I need a sample business plan for opening a Histology lab (human tissue in Dallas Texas); basically focusing on technical and surgical dermatological samples where its going to involve all aspects in Histology from mohs,special stains,immunos,mycology,embedding, cutting, staining etc... so I can have an idea in how to plan or write, specially the projection part is given me trouble. I got all the outline from the Small Business Bureau on how to write it but I need a little help by sending me a sample plan please, I could not find any online. Any suggestions! Thanks for your time and attention on this matter. Ali Histotechnologist ____________________________________________________________________________________ Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. http://overview.mail.yahoo.com/ From AnthonyH <@t> chw.edu.au Fri Nov 23 20:11:50 2007 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Nov 23 20:13:49 2007 Subject: [Histonet] 35 mm transfer device Message-ID: You can also use these scanners (with a 35mm slide adapter) to scan whole slides This comes in useful when a whole-mount is required. 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: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of jstaruk Sent: Saturday, 24 November 2007 7:17 AM To: 'Richard Cartun'; 'Histonet' Subject: RE: [Histonet] 35 mm transfer device My Epson Stylus CX7800 printer / scanner came with a neat little device that scans (I think) 6 at a time and saves them as individual digital images. I transferred all of my Kodachromes to digital this way. They look great! Jim _____________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Friday, November 23, 2007 3:06 PM To: Histonet Subject: [Histonet] 35 mm transfer device Does anyone have a recommendation for a device that will transfer "old" 35 mm color slides into "modern day" digital images? 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************* This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note 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 dis.tox <@t> suven.com Sat Nov 24 03:48:40 2007 From: dis.tox <@t> suven.com (Toxicology) Date: Sat Nov 24 03:42:14 2007 Subject: [Histonet] Liver cells Message-ID: Can anyone help me out to recognize these liver cells. I have pasted image on histonet as LIVER Vinod -------------------------Email Disclaimer--------------------------------- This e-mail may contain confidential and/or privileged information. If you are not the intended recipient (or have received this e-mail in error) please notify the sender immediately and destroy this e-mail. Any unauthorized copying, disclosure or distribution of the material in this e-mail is strictly forbidden. From carl.hobbs <@t> kcl.ac.uk Sat Nov 24 12:49:25 2007 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Sat Nov 24 12:49:50 2007 Subject: [Histonet] re: 35 mm transfer device Message-ID: <001001c82eca$bc733f00$4101a8c0@carlba65530bda> I agree with Gail. I use a dedicated Nikon 35mm scanner for work. I have at home one too! Canon's is just as good.....cheaper, I recall. However, at home I tend to use my Epson Perfection 4870 flatbed scanner for most of my non-essential 35mm conversions. Sure, it has a much bigger footprint but it does transmitted/reflective scans. I used to be a Umax fan but no longer. NB: a good image processing software app is very important, as Gail pointed out. Imho, one needs to buy Adobe's Photoshop, if you are "professionally inclined". For home use, Adobe's "Elements" is excellent...so is the free Irfanview. In fact , I use Irfanview for many image resizings. Great respect for The Man behind "Irfanview". C From kevinclayton1979 <@t> gmail.com Mon Nov 26 08:32:39 2007 From: kevinclayton1979 <@t> gmail.com (Kevin Clayton) Date: Mon Nov 26 08:32:53 2007 Subject: [Histonet] quantifying only the surface-expressed levels of a receptor in sections Message-ID: <94c6d26c0711260632w563d9476gbdac300f8e6dafed@mail.gmail.com> Hi all As the subject says, I want to quantify only the surface levels of glutamate receptors in mouse brain vibratome sections. I want to see if there are more/less receptors from mice that have received a certain treatment. Now that I have the antibodies working well I realise that it may not even be possible to specifically look at the level of the receptors in the cell membrane (excluding those in intracellular vesicles, for example). Is it possible? I don't permeabilize of course but sectioning itself must damage the integrity of the cells. Or can that be overcome by only taking confocal slices from further in the tissue? Thanks Kevin From kevinclayton1979 <@t> gmail.com Mon Nov 26 09:14:27 2007 From: kevinclayton1979 <@t> gmail.com (Kevin Clayton) Date: Mon Nov 26 09:14:39 2007 Subject: [Histonet] Update: quantifying only the surface-expressed levels of a receptor in sections Message-ID: <94c6d26c0711260714i73786d3ak826794de00e71674@mail.gmail.com> Sorry to mail again but I just noticed that as the brains were cryoprotected in glycerol/ethylene glycol/PBS they've already been permeabilized (right?). Kevin -------------------------- Hi all As the subject says, I want to quantify only the surface levels of glutamate receptors in mouse brain vibratome sections. I want to see if there are more/less receptors from mice that have received a certain treatment. Now that I have the antibodies working well I realise that it may not even be possible to specifically look at the level of the receptors in the cell membrane (excluding those in intracellular vesicles, for example). Is it possible? I don't permeabilize of course but sectioning itself must damage the integrity of the cells. Or can that be overcome by only taking confocal slices from further in the tissue? Thanks Kevin From ian.montgomery <@t> bio.gla.ac.uk Mon Nov 26 09:33:28 2007 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Mon Nov 26 09:33:49 2007 Subject: [Histonet] Fish brain. Message-ID: <003701c83041$b19f8c50$6424d182@IBLS.GLA.AC.UK> I'm about to start a general study on brown trout brain. In size it's similar to a mouse brain so I intend using a processing schedule that suits mouse brain. But, it's fish brain, any hints and tips? Ian. Dr. Ian Montgomery, Histotechnology, I.B.L.S. Support Unit, Thomson Building, University of Glasgow, G12 8QQ. From rjbuesa <@t> yahoo.com Mon Nov 26 09:59:54 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Nov 26 10:00:10 2007 Subject: [Histonet] Fish brain. In-Reply-To: <003701c83041$b19f8c50$6424d182@IBLS.GLA.AC.UK> Message-ID: <764050.60158.qm@web61220.mail.yahoo.com> It is not as "dry" as mouse brain (and not because it is a fish!). Ren? J. Ian Montgomery wrote: I'm about to start a general study on brown trout brain. In size it's similar to a mouse brain so I intend using a processing schedule that suits mouse brain. But, it's fish brain, any hints and tips? Ian. Dr. Ian Montgomery, Histotechnology, I.B.L.S. Support Unit, Thomson Building, University of Glasgow, G12 8QQ. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From MadaryJ <@t> MedImmune.com Mon Nov 26 10:48:41 2007 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Mon Nov 26 10:49:19 2007 Subject: [Histonet] need cd19 help in mouse spleens and a job Message-ID: <8F3E1865E343C943BB38506D56FF015101335516@MD1EV002.medimmune.com> I need help finding a source and even a decent procedure that works on cd19 on FFPE mouse spleens. I have a decent procedure for cd3 and cd45, but the path wants cd19, so I figured I would throw it to the experts first. Thanks in advance. Also Medimmune is looking for a histotech here in Rockville Md. Someone with GLP, frozen and IHC experience. Nick Madary, HT/HTL(ASCP)QIHC Histology Mgr, Medimmune 301.398.6360(lab), 4745(vm),9745(fax) "To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary, and expected to be used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation." From kmilne <@t> bccancer.bc.ca Mon Nov 26 12:11:54 2007 From: kmilne <@t> bccancer.bc.ca (Milne, Katy) Date: Mon Nov 26 12:15:46 2007 Subject: [Histonet] RE: need cd19 help in mouse spleens and a job References: <6e37ab$25la17@ironport1in.cw.bc.ca> Message-ID: <07979E76B0869D4E8C9FE4AA9FC06578031E6C7F@srvex03.phsabc.ehcnet.ca> Hi Nick, I wasn't very successful at finding a CD19 Ab for mouse FFPE but I did find a PAX5 Ab that worked for detecting B cells, it came from Lab Vision (RB-9406), it should give nuclear staining in B cells but not in plasma cells though. You could potentially try CD79a for that as I think that comes up strong in plasma cells but again getting a good Ab for mouse FFPE for that is tricky. Katy Milne Deeley Research Centre Message: 5 Date: Mon, 26 Nov 2007 11:48:41 -0500 From: "Madary, Joseph" Subject: [Histonet] need cd19 help in mouse spleens and a job To: Message-ID: <8F3E1865E343C943BB38506D56FF015101335516@MD1EV002.medimmune.com> Content-Type: text/plain; charset="us-ascii" I need help finding a source and even a decent procedure that works on cd19 on FFPE mouse spleens. I have a decent procedure for cd3 and cd45, but the path wants cd19, so I figured I would throw it to the experts first. Thanks in advance. Also Medimmune is looking for a histotech here in Rockville Md. Someone with GLP, frozen and IHC experience. Nick Madary, HT/HTL(ASCP)QIHC Histology Mgr, Medimmune 301.398.6360(lab), 4745(vm),9745(fax) "To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary, and expected to be used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation." ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 36 **************************************** From fudo <@t> ufl.edu Mon Nov 26 12:35:56 2007 From: fudo <@t> ufl.edu (FU,DONGTAO) Date: Mon Nov 26 12:36:02 2007 Subject: [Histonet] Dewax solution Message-ID: <1599229135.303891196102156131.JavaMail.osg@osgjas04.cns.ufl.edu> Hi all, We usually use Xylene to dewax paraffin. Now we want to get away from using xylene at some point and use heat-based dewaxing instead. Does anyone have some experiences on it? Which company's product do you use? Many thanks, Ann Dongtao Fu MD, Ph.D Lab Manager Molecular Pathology Core Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 From mtarango <@t> nvcancer.org Mon Nov 26 12:35:49 2007 From: mtarango <@t> nvcancer.org (Tarango, Mark) Date: Mon Nov 26 12:36:11 2007 Subject: [Histonet] need cd19 help in mouse spleens and a job In-Reply-To: <8F3E1865E343C943BB38506D56FF015101335516@MD1EV002.medimmune.com> Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6FDA@NVCIEXCH02.NVCI.org> BioCare recently came out with an anti-CD19 antibody that works in FFPE tissue sections. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Madary, Joseph Sent: Monday, November 26, 2007 8:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] need cd19 help in mouse spleens and a job I need help finding a source and even a decent procedure that works on cd19 on FFPE mouse spleens. I have a decent procedure for cd3 and cd45, but the path wants cd19, so I figured I would throw it to the experts first. Thanks in advance. Also Medimmune is looking for a histotech here in Rockville Md. Someone with GLP, frozen and IHC experience. Nick Madary, HT/HTL(ASCP)QIHC Histology Mgr, Medimmune 301.398.6360(lab), 4745(vm),9745(fax) "To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary, and expected to be used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation." _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== From fudo <@t> ufl.edu Mon Nov 26 12:36:08 2007 From: fudo <@t> ufl.edu (FU,DONGTAO) Date: Mon Nov 26 12:36:30 2007 Subject: [Histonet] Dewax solution Message-ID: <1428458827.303941196102168921.JavaMail.osg@osgjas04.cns.ufl.edu> Hi all, We usually use Xylene to dewax paraffin. Now we want to get away from using xylene at some point and use heat-based dewaxing instead. Does anyone have some experiences on it? Which company's product do you use? Many thanks, Ann Dongtao Fu MD, Ph.D Lab Manager Molecular Pathology Core Dept. of Pathology Lab phone: 352-273-7752 Lab FAX: 352-273-7755 Lab address: D11-50 PO Box: 100275 1600 SW Archer Road University of Flodrida Gainesville, FL 32610 From Heather.D.Renko <@t> osfhealthcare.org Mon Nov 26 13:21:24 2007 From: Heather.D.Renko <@t> osfhealthcare.org (Renko, Heather D.) Date: Mon Nov 26 13:21:41 2007 Subject: [Histonet] sentinel biopsies Message-ID: <40026EDDE64CDA47AB382C52619ACD3C0A09DC27@pmc-rfd-mx01.intranet.osfnet.org> Hazel, I seen your query a few years back on your sentinal node protocol and did not know if your obtained your information on how to dispose of radioactive material from your nodes. If so, can I see your protocols. I am revising mine and would appreciate any help available. Heather D. Renko, Histology Coordinator OSF Saint Anthony Medical Center 5666 East State Street Rockford, Illinois 61108 Direct: 815-395-5410 Heather.D.Renko@osfhealthcare.org ============================================================================== The information in this message is confidential and may be legally privileged. Access to this message by anyone other than the addressee is not authorized. If you are not the intended recipient, or an agent of the intended recipient, any disclosure, copying, or distribution of the message or any action or omission taken by you in reliance on it, is prohibited and may be unlawful. If you have received this message in error, please contact the sender immediately and permanently delete the original e-mail, attachment(s), and any copies. ============================================================================== From SHargrove <@t> urhcs.org Mon Nov 26 16:53:19 2007 From: SHargrove <@t> urhcs.org (SHargrove@urhcs.org) Date: Mon Nov 26 16:53:37 2007 Subject: [Histonet] Susie Hargrove is out of the office. Message-ID: I will be out of the office starting 11/26/2007 and will not return until 12/04/2007. I will respond to your message when I return. If immediate assistance is needed please call 3198. From sbreeden <@t> nmda.nmsu.edu Tue Nov 27 10:00:17 2007 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Tue Nov 27 10:00:24 2007 Subject: [Histonet] SelecTech Staining Message-ID: <4D14F0FC9316DD41972D5F03C070908B017E609F@nmdamailsvr.nmda.ad.nmsu.edu> Is anyone out there using Surgipath's SelecTech stains (Blue Buffer and Define) with a regressive hematoxylin on a LEICA AUTOSTAINER XL? If so, would you be willing to offer your staining protocol (setup and times)? Thanks so much! Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 From gvdobbin <@t> ihis.org Tue Nov 27 11:03:40 2007 From: gvdobbin <@t> ihis.org (Greg Dobbin) Date: Tue Nov 27 11:04:17 2007 Subject: [Histonet] SelecTech Staining Message-ID: I am using them on the Dako Autostainer XL and sure I'll share! Here is our schedule: Oven, 65C 10:00 Xylene 3 min Xylene 2 min Abs. ETOH 1 min Abs. ETOH 1 min 95% ETOH 1 min 70% ETOH 1 min Wash, Tap H2O 2 min Wash, dist. H2O 1 min Hematox. 560 4 min Wash, Tap H2O 2 min Define 1 min Wash, Tap H2O 2 min Bluing 1 min Wash, Tap H2O 70% ETOH 1 min Alc. Eosin 515 30 sec Abs. ETOH 1 min Abs. ETOH 1 min Abs. ETOH 1 min Xylene 2 min Xylene 2 min Exit We (the technologists) love the these reagents-no more filtering, and the docs love the staining. We have toyed with the idea of adding a 50% ETOH step after eosin to reduce the eosin intensity, but we would have to lose another station and the docs are happy with it as it is, sooo for now we will not be bothered. Good luck with them. Greg Greg Dobbin, R.T. Chief Technologist, Histology Lab Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 There is some merit in doing the right thing rather badly, but absolutely none in doing the wrong thing excellently! >>> "Breeden, Sara" 11/27/2007 12:00 PM >>> Is anyone out there using Surgipath's SelecTech stains (Blue Buffer and Define) with a regressive hematoxylin on a LEICA AUTOSTAINER XL? If so, would you be willing to offer your staining protocol (setup and times)? Thanks so much! Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. D?claration de confidentialit? Le pr?sent message (y compris les annexes) peut contenir des renseignements confidentiels ayant pour objet une personne ou un organisme particulier. Si vous avez re?u la pr?sente communication par erreur, veuillez en informer l'exp?diteur imm?diatement. Si vous n'?tes pas le destinataire pr?vu, vous n'avez pas le droit d'utiliser, divulguer, distribuer, copier ou imprimer ce courriel ou encore de vous en servir, et vous devriez l'effacer compl?tement de votre syst?me informatique. From jennifer.harvey <@t> Vanderbilt.Edu Tue Nov 27 11:28:27 2007 From: jennifer.harvey <@t> Vanderbilt.Edu (Harvey, Jennifer Lynn) Date: Tue Nov 27 11:28:33 2007 Subject: [Histonet] NaCl in antibody diluent Message-ID: Histonet, Has anyone tried NaCl in there diluent buffer for IHC, if so what concentration. I read about it in DAKO's immunostaining methods booklet but they don't state the concentration. This is my last chance with my background problem. I have a sticky collagen issue I cant get rid of. Thanks! Jennifer Harvey, HT(ASCP) QIHC Vanderbilt Vision Research Center RM 8105 MCE North Tower 1215 21st Ave. South Nashville, TN 37232-8808 Phone: 615-936-1486 From trathborne <@t> somerset-healthcare.com Tue Nov 27 11:33:06 2007 From: trathborne <@t> somerset-healthcare.com (Rathborne, Toni) Date: Tue Nov 27 11:33:28 2007 Subject: [Histonet] Control tissue for PCP Message-ID: Hi Histonetters. Some time ago, I read a procedure for growing your own Gram +/- controls using a piece of fresh liver. Has anyone tried something similar with lung tissue for PCP? The control slides are very expensive, so we're looking for an alternative. Thanks for any ideas you may have. Toni CONFIDENTIALITY NOTICE This message and any included attachments are from Somerset Medical Center and are intended only for the addressee. The information contained in this message is confidential and may contain privileged, confidential, proprietary and/or trade secret information entitled to protection and/or exemption from disclosure under applicable law. Unauthorized forwarding, printing, copying, distribution, or use of such information is strictly prohibited and may be unlawful. If you are not the addressee, please promptly delete this message and notify the sender of the delivery error by e-mail or you may call Somerset Medical Center's computer Help Desk at 908-685-2200, ext. 4050. Somerset Medical Center is proud to receive the Somerset County Business Partnership's 2006 Quality of Life Award. From talulahgosh <@t> gmail.com Tue Nov 27 11:42:34 2007 From: talulahgosh <@t> gmail.com (Emily Sours) Date: Tue Nov 27 11:42:39 2007 Subject: [Histonet] vibratome circulating bath Message-ID: We purchased a vibratome which comes with a buffer ice tray. Does this serve the same purpose as the circulating bath (to keep the buffer cold)? Why would one need a circulating bath? Emily -- crow tom mike and joel trapped in space the endless void we've got movie sign! mst3k haiku brought to you by joe wiencis From rjbuesa <@t> yahoo.com Tue Nov 27 11:44:33 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 27 11:44:37 2007 Subject: [Histonet] NaCl in antibody diluent In-Reply-To: Message-ID: <244249.10617.qm@web61211.mail.yahoo.com> Jennifer: The Ab diluent is made of 1% bovine serum albumen in PBS with ).05% Tween 20 and 0.1% sodium azide.'NaCl is part of the PBS but I have never used or kbown anybody using additional NaCl Ren? J. "Harvey, Jennifer Lynn" wrote: Histonet, Has anyone tried NaCl in there diluent buffer for IHC, if so what concentration. I read about it in DAKO's immunostaining methods booklet but they don't state the concentration. This is my last chance with my background problem. I have a sticky collagen issue I cant get rid of. Thanks! Jennifer Harvey, HT(ASCP) QIHC Vanderbilt Vision Research Center RM 8105 MCE North Tower 1215 21st Ave. South Nashville, TN 37232-8808 Phone: 615-936-1486 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Never miss a thing. Make Yahoo your homepage. From ttroyer <@t> petersonlab.com Tue Nov 27 11:54:57 2007 From: ttroyer <@t> petersonlab.com (Travis Troyer) Date: Tue Nov 27 11:47:56 2007 Subject: [Histonet] PGP 9.5 Message-ID: <002b01c8311e$a053b740$6e01010a@Peterson.local> I was asked today about PGP 9.5 (Protein Gene Product 9.5) and was wondering if anyone has heard of a company that can do this as a clinical test. Thanks, Travis From rjbuesa <@t> yahoo.com Tue Nov 27 11:48:01 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 27 11:48:07 2007 Subject: [Histonet] vibratome circulating bath In-Reply-To: Message-ID: <319503.37631.qm@web61219.mail.yahoo.com> A circulating bath is the best solution to keeping any liquid temperature within narrow limits because the liquid is circulated through a chiller (or heater, depending of the temperature you want to maintain) and compensate for the thermal variations caused by the environment on a still liquid. Ren? J. Emily Sours wrote: We purchased a vibratome which comes with a buffer ice tray. Does this serve the same purpose as the circulating bath (to keep the buffer cold)? Why would one need a circulating bath? Emily -- crow tom mike and joel trapped in space the endless void we've got movie sign! mst3k haiku brought to you by joe wiencis _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. From TJJ <@t> Stowers-Institute.org Tue Nov 27 12:15:56 2007 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Tue Nov 27 12:16:21 2007 Subject: [Histonet] Re: NaCl in antibody diluent Message-ID: Jennifer, I used NaCl years ago in a clinical lab environment. We never put it in antibody diluent. We did use it in our buffer rinses though, at a final concentration of 0.9% (normal saline). It was thought to reduce background staining. I don't know if it did in any great measure, but we kept using it because it didn't hurt to have it in there. Since then we have moved to automation and there is not any NaCl in our buffers. Are you getting non-specific binding in your collagen with all your antibodies, or just a couple? Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 From RSRICHMOND <@t> aol.com Tue Nov 27 12:17:36 2007 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Tue Nov 27 12:17:41 2007 Subject: [Histonet] Re: sentinel biopsies Message-ID: Heather D. Renko, Histology Coordinator, OSF Saint Anthony Medical Center, Rockford Illinois asks: >>I seen your query a few years back on your sentinel node protocol and did not know if your obtained your information on how to dispose of radioactive material from your nodes. If so, can I see your protocols. I am revising mine and would appreciate any help available.<< The radioisotope used in sentinel node procedures is technetium 99m, which has a half-life of six hours. The level of radioactivity of these specimens is low enough that they do not require any special handling. By the time a pathology specimen is ready to be disposed of (at least a week, probably more) the radioisotope is many half-lives out, and the level of radiation probably undetectable. Bob Richmond Samurai Pathologist Knoxville TN From failm <@t> musc.edu Tue Nov 27 12:43:18 2007 From: failm <@t> musc.edu (Fail, Mildred M.) Date: Tue Nov 27 12:44:42 2007 Subject: [Histonet] Cutting nails Message-ID: I am trying to compare techniques for cutting nails. What do you use for sofenting nails before processing and/or cutting? Detergent, nair, alkaline solution? How long are they placed in the solution? Thank you Rena Fail From BSauer <@t> stjosephswb.com Tue Nov 27 13:10:29 2007 From: BSauer <@t> stjosephswb.com (Sauer, Barb) Date: Tue Nov 27 13:10:42 2007 Subject: [Histonet] Iron stains on smears Message-ID: Does anyone have experience with doing iron stains on smears that have been prepared with blood from heparinized tubes? We are having trouble with our iron stain for bone marrow cases since we changed to heparinized tubes and thought that could be the problem. Thanks for your help ******************************************* This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the system manager. ******************************************* From rchiovetti <@t> yahoo.com Tue Nov 27 13:14:16 2007 From: rchiovetti <@t> yahoo.com (Robert Chiovetti) Date: Tue Nov 27 13:14:20 2007 Subject: [Histonet] NaCl in antibody diluent Message-ID: <70716.80367.qm@web58902.mail.re1.yahoo.com> Jennifer, I can't speak to IHC, but I've done quite a bit of immunolabeling for electron microscopy. We routinely used NaCl at 0.9% (by weight) in diluents and washes with pretty good success. That's considered to be "normal saline," a physiologically relevant concentration. 0.9% by weight is 150 mM, if I recall correctly. We also used to use *very* high NaCl concentrations in our washes when we had background problems, and the high salt would frequently get rid of the background staining. In those cases we'd use about 5X normal saline (4.5% NaCl by weight) as the rinse after the primary and secondary Ab's. You just have to remember to return to 0.9% normal saline for a couple of washes before continuing with the labeling procedure. Don't ask me to explain why this worked, but it frequently cleaned up messy background problems. A biochemist friend of mine says this is similar to what he does when he cleans and preps his gel columns after using them. He uses high salt to remove traces of proteins and contaminants from the columns. The way he explains it, the high salt makes most proteins completely collapse so they're no longer "sticky" and they can't bind to anything. The strange thing is, the specific Ab-Ag binding seemed to survive the high salt just fine. And my disclaimer: This frequently (but not always) worked for immunoEM. I have no experience with IHC, but it might be worth a try... Cheers, Bob Robert (Bob) Chiovetti, Ph.D. Southwest Precision Instruments The Desert Southwest's Microscopy Resource www.swpinet.com 132 North Elster Drive Tucson, AZ 85710-3212 Tel./Fax 520-546-4986 Member, Arizona Small Business Association (www.asba.com) ----- Original Message ---- Has anyone tried NaCl in there diluent buffer for IHC, if so what concentration. I read about it in DAKO's immunostaining methods booklet but they don't state the concentration. This is my last chance with my background problem. I have a sticky collagen issue I cant get rid of. ____________________________________________________________________________________ Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. http://overview.mail.yahoo.com/ From Wanda.Smith <@t> HCAhealthcare.com Tue Nov 27 13:30:48 2007 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Tue Nov 27 13:30:57 2007 Subject: [Histonet] Re: sentinel biopsies In-Reply-To: References: Message-ID: <817C2761C5A1394180709AEEDB775B7E0404092E@NASEV03.hca.corpad.net> I agree, that by the time it is time to dispose of the specimen (after 2 weeks per CAP), they do not need any special handling. We hold the s. node and breast for 24 hrs for fixation, however, we are finding on some cases we have to hold the sentinel node an extra day due to the level of radioactivity when we check it with the Geiger counter. Is anyone else having this problem? Thanks, Wanda Wanda G. Smith, HTL(ASCP)HT Trident Medical Center Pathology Supervisor ph: 843-847-4586 fx: 843-847-4296 email: wanda.smith@hcahealthcare.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert Richmond Sent: Tuesday, November 27, 2007 1:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: sentinel biopsies Heather D. Renko, Histology Coordinator, OSF Saint Anthony Medical Center, Rockford Illinois asks: >>I seen your query a few years back on your sentinel node protocol and did not know if your obtained your information on how to dispose of radioactive material from your nodes. If so, can I see your protocols. I am revising mine and would appreciate any help available.<< The radioisotope used in sentinel node procedures is technetium 99m, which has a half-life of six hours. The level of radioactivity of these specimens is low enough that they do not require any special handling. By the time a pathology specimen is ready to be disposed of (at least a week, probably more) the radioisotope is many half-lives out, and the level of radiation probably undetectable. Bob Richmond Samurai Pathologist Knoxville TN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RSRICHMOND <@t> aol.com Tue Nov 27 14:12:54 2007 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Tue Nov 27 14:13:01 2007 Subject: [Histonet] Re: sentinel biopsies In-Reply-To: <817C2761C5A1394180709AEEDB775B7E0404092E@NASEV03.hca.corpad.net> References: <817C2761C5A1394180709AEEDB775B7E0404092E@NASEV03.hca.corpad.net> Message-ID: Wanda G. Smith at Trident Medical Center (where?) notes: >>I agree, that by the time it is time to dispose of the specimen (after 2 weeks per CAP), they do not need any special handling. We hold the sentinel node and breast for 24 hours for fixation, however, we are finding on some cases we have to hold the sentinel node an extra day due to the level of radioactivity when we check it with the Geiger counter. Is anyone else having this problem?<< Once again - the authorities on this subject agree - this is not my personal opinion alone - that the level of radioactivity in these specimens does not require any special handling. You should cut the specimen, fix it overnight (for histological reasons), and process it routinely. 24 hours equals four half-lives of technetium 99m, and I would expect you could still detect radioactivity after that time. If your radiation safety people have any further misgivings, I think this topic has been discussed on Histonet before, and that literature citations were given. If this information isn't available in our archives, I can probably look it up. Bob Richmond Samurai Pathologist Knoxville TN From rjbuesa <@t> yahoo.com Tue Nov 27 14:32:46 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Nov 27 14:32:52 2007 Subject: [Histonet] Cutting nails In-Reply-To: Message-ID: <949558.89406.qm@web61220.mail.yahoo.com> What was worked for me consistently as better, is placing the fixed nail in a 10% solution of NaOH for half an hour, wash thoroughly and process as usual. Ren? J. "Fail, Mildred M." wrote: I am trying to compare techniques for cutting nails. What do you use for sofenting nails before processing and/or cutting? Detergent, nair, alkaline solution? How long are they placed in the solution? Thank you Rena Fail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From c.m.vanderloos <@t> amc.uva.nl Wed Nov 28 01:41:27 2007 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Wed Nov 28 01:41:38 2007 Subject: [Histonet] RE: NaCl in antibody diluent Message-ID: <103e2610704b.10704b103e26@amc.uva.nl>

Jennifer and Rene,

As Rene said the NaCl is part of the PBS. The 0.9% concentra 0.15 M. I have heard (but never tested) of somebody the NaCl concentration to 0.5 M for reducing non-specific ba ckground staining. He used this high salt PBS not only for diluting the concent certainly destroy

Cheers,

Chris van der Loos, PhD
Dept. of Pathology
Acade M2-230
Meibergdreef 9
NL-1105 AZ Amsterd Netherlands

 

Date: Tue, 27 Nov 2007 09:44:33 -0800 (PST)
From: <rjbuesa@yahoo.com>
Subject: Re: [Histonet] NaCl in antibody diluent
To: "Harvey, Jennif <jennifer.harvey@Vanderbilt.Edu>,
histonet@lists.utsouthwestern.edu

Jennifer: in PBS wit is part of the P additional NaCl
& J.

"Harvey, Jennifer Lynn" < jennifer.harvey@Vanderbilt.Edu> wrote:
  H tried NaCl in there diluent buffer f what
concentration. I read about it in DAKO's immunostaining methods booklet
but they don't state the concentr problem. I ha of.

Thanks!< From Tony_Reilly <@t> health.qld.gov.au Wed Nov 28 02:13:19 2007 From: Tony_Reilly <@t> health.qld.gov.au (Anthony Reilly) Date: Wed Nov 28 02:13:34 2007 Subject: [Histonet] NaCl in antibody diluent Message-ID: Hi NaCl is a normal component of PBS. The high salt buffer DAKO are most likely referring to is TBS which is a Tris buffer with 0.9% NaCl added. As others have stated this is used to reduce background. In my experience this will only give minimal improvement. If you are suffering background you are best to investigate your dilution, blocking if any and the type of detection system used. regards Tony Reilly Chief Scientist Anatomical Pathology Pathology Queensland Prince Charles Hospital Rode Rd Chermside Q 4032 Australia Ph: 07 3139 4543 Fax: 07 3193 4546 tony_reilly@health.qld.gov.au >>> "Harvey, Jennifer Lynn" 11/28/07 3:28 am >>> Histonet, Has anyone tried NaCl in there diluent buffer for IHC, if so what concentration. I read about it in DAKO's immunostaining methods booklet but they don't state the concentration. This is my last chance with my background problem. I have a sticky collagen issue I cant get rid of. Thanks! Jennifer Harvey, HT(ASCP) QIHC Vanderbilt Vision Research Center RM 8105 MCE North Tower 1215 21st Ave. South Nashville, TN 37232-8808 Phone: 615-936-1486 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/ received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is strictly prohibited. The information contained in this email, including any attachment sent with it, may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone collect on Australia +61 1800 198 175 or by return email. You should also delete this email, and any copies, from your computer system network and destroy any hard copies produced. If not an intended recipient of this email, you must not copy, distribute or take any action(s) that relies on it; any form of disclosure, modification, distribution and/or publication of this email is also prohibited. Although Queensland Health takes all reasonable steps to ensure this email does not contain malicious software, Queensland Health does not accept responsibility for the consequences if any person's computer inadvertently suffers any disruption to services, loss of information, harm or is infected with a virus, other malicious computer programme or code that may occur as a consequence of receiving this email. Unless stated otherwise, this email represents only the views of the sender and not the views of the Queensland Government. **************************************************************** From nfournier <@t> sasktel.net Wed Nov 28 03:17:56 2007 From: nfournier <@t> sasktel.net (Neil Fournier) Date: Wed Nov 28 03:17:34 2007 Subject: [Histonet] substance P receptor Message-ID: <000801c8319f$903347a0$839b2fcf@NEIL> Does anybody have experience staining rat brain for the substance P receptor? Which company would you recommend? I have also read that many researchers fix tissue with paraformaldehyde and picric acid? Unfortunately, when we designed our experiments we did not anticipate that we would have to stain our tissue with this antibody and I was wondering how important is it to fix tissue with a paraformaldehyde/picric acid solution. Our rats were perfused with physiological saline followed by 4% paraformaldehyde. Brains were removed and postfixed for 72 hrs in the same solution, before being placed in 0.1% sodium azide/PB solution prior to sectioning on a Vibratome at 50 micron. Thanks for the help, Neil From M.Walker <@t> hrsu.mrc.ac.uk Wed Nov 28 09:50:18 2007 From: M.Walker <@t> hrsu.mrc.ac.uk (Marion Walker) Date: Wed Nov 28 09:50:43 2007 Subject: [Histonet] Xylene-free processing & immunology [Scanned] In-Reply-To: References: Message-ID: <86F334797DC6524A99AD9DD8F23A8B500DDD2E@mailserv.hrsu.mrc.ac.uk> Has anybody tried any immuno staining on xylene free paraffin processed samples? Did it work? All the reference that I am coming across refer to histological stainings but nothing about immunos. Arantza Esnal.MRC-HRSU, Edinburgh. From rjbuesa <@t> yahoo.com Wed Nov 28 10:06:42 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 28 10:06:50 2007 Subject: [Histonet] Xylene-free processing & immunology [Scanned] In-Reply-To: <86F334797DC6524A99AD9DD8F23A8B500DDD2E@mailserv.hrsu.mrc.ac.uk> Message-ID: <78101.92531.qm@web61218.mail.yahoo.com> My xylene free (with mineral oil) tissue processing was tested for all the IHC procedures we used at the moment. If you are interested I can send you an article I published on the subject. Ren? J. Marion Walker wrote: Has anybody tried any immuno staining on xylene free paraffin processed samples? Did it work? All the reference that I am coming across refer to histological stainings but nothing about immunos. Arantza Esnal.MRC-HRSU, Edinburgh. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. From rhrogers <@t> iupui.edu Wed Nov 28 10:14:51 2007 From: rhrogers <@t> iupui.edu (Rogers, Rhonda) Date: Wed Nov 28 10:15:00 2007 Subject: [Histonet] benzidine stain to demonstrate red blood cells in whole embryos Message-ID: <79188A7B37805441AB277C7F7B7F9A8F0812EAF35F@iu-mssg-mbx03.ads.iu.edu> Good morning, I am looking for a method to stain whole mouse embryos with benzidine to demonstrate red blood cells. The one we were given by a colleague is not working for us, so I hope someone on Histonet is willing to share a reliable method. Regards, Rhonda Rhonda Rogers, HTL(ASCP) Wells Center for Pediatric Research - Conway Laboratory Indiana University Purdue University Indianapolis 1044 W. Walnut St., R4 379 Indianapolis, IN 46202-5225 Phone: 317-278-8781 E-mail: rhrogers@iupui.edu From gu.lang <@t> gmx.at Wed Nov 28 10:31:45 2007 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Wed Nov 28 10:31:53 2007 Subject: AW: [Histonet] Xylene-free processing & immunology [Scanned] In-Reply-To: <86F334797DC6524A99AD9DD8F23A8B500DDD2E@mailserv.hrsu.mrc.ac.uk> Message-ID: <000301c831dc$2b2bdc40$6412a8c0@dielangs.at> Our tissue is routinly processed with shellsol instead of xylene for decades. It's a kind of mineral spirit, has no influence on immunhisto. Gudrun Lang Biomed. Analytikerin Histolabor Akh Linz Krankenhausstr. 9 4020 Linz +43(0)732/7806-6754 -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Marion Walker Gesendet: Mittwoch, 28. November 2007 16:50 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Xylene-free processing & immunology [Scanned] Has anybody tried any immuno staining on xylene free paraffin processed samples? Did it work? All the reference that I am coming across refer to histological stainings but nothing about immunos. Arantza Esnal.MRC-HRSU, Edinburgh. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed Nov 28 10:33:54 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Nov 28 10:33:59 2007 Subject: [Histonet] benzidine stain to demonstrate red blood cells in whole embryos In-Reply-To: <79188A7B37805441AB277C7F7B7F9A8F0812EAF35F@iu-mssg-mbx03.ads.iu.edu> Message-ID: <738079.60914.qm@web61216.mail.yahoo.com> And you should be glad it does not work, because benzidine is one of the most dangerous substances you can work with. Stay away from it! Ren? J. "Rogers, Rhonda" wrote: Good morning, I am looking for a method to stain whole mouse embryos with benzidine to demonstrate red blood cells. The one we were given by a colleague is not working for us, so I hope someone on Histonet is willing to share a reliable method. Regards, Rhonda Rhonda Rogers, HTL(ASCP) Wells Center for Pediatric Research - Conway Laboratory Indiana University Purdue University Indianapolis 1044 W. Walnut St., R4 379 Indianapolis, IN 46202-5225 Phone: 317-278-8781 E-mail: rhrogers@iupui.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. From derek.papalegis <@t> tufts.edu Wed Nov 28 10:34:01 2007 From: derek.papalegis <@t> tufts.edu (Derek Papalegis) Date: Wed Nov 28 10:34:08 2007 Subject: [Histonet] benzidine stain to demonstrate red blood cells in whole embryos In-Reply-To: <79188A7B37805441AB277C7F7B7F9A8F0812EAF35F@iu-mssg-mbx03.ads.iu.edu> References: <79188A7B37805441AB277C7F7B7F9A8F0812EAF35F@iu-mssg-mbx03.ads.iu.edu> Message-ID: <474D9879.1020507@tufts.edu> Rogers, Rhonda wrote: > Good morning, > > I am looking for a method to stain whole mouse embryos with benzidine to demonstrate red blood cells. The one we were given by a colleague is not working for us, so I hope someone on Histonet is willing to share a reliable method. > > Regards, > > Rhonda > > > Rhonda Rogers, HTL(ASCP) > > Wells Center for Pediatric Research - Conway Laboratory > > Indiana University Purdue University Indianapolis > > 1044 W. Walnut St., R4 379 > > Indianapolis, IN 46202-5225 > > > > Phone: 317-278-8781 E-mail: rhrogers@iupui.edu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Hi Rhonda, I recently had an investigator request this. We got the best results by staining the mouse embryos with Benzadine after fixation in Bouin's. I believe we stained them for 30 minutes in Benzadine. After staining, just process and cut like normal and you should see staining on the tissue. In addition to the Benzadine staining, we stained the slides with Okajima's and the investigator was very happy with the results. I hope this helps, Derek -- Derek Papalegis HT (ASCP) Histotechnician Division of Laboratory Animal Medicine Tufts University 136 Harrison Avenue Boston, MA 02111 phone: 617 636-2971 fax: 617 636-8354 From am3309 <@t> uga.edu Wed Nov 28 11:29:43 2007 From: am3309 <@t> uga.edu (Abigail M. Butler) Date: Wed Nov 28 11:29:49 2007 Subject: [Histonet] Pay Range Message-ID: <20071128122943.JPR06067@punts1.cc.uga.edu> Maybe someone can help me out.... I am just curious to know what other histotechs are making that have the same duties as I do. I am HT certified, QIHC qualified, I do ALL the IHC in the lab, I do ordering, I do all the monthly billing, and routine stuff as well. I am not the supervisor just a tech. Can anyone offer any salaries? Thanks Abbie Butler, HT (ASCP), QIHC University of Georgia College of Veterinary Medicine From sjchtascp <@t> yahoo.com Wed Nov 28 12:07:22 2007 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Wed Nov 28 12:07:25 2007 Subject: [Histonet] HT position Wanted Madison, WI Message-ID: <484534.46069.qm@web38204.mail.mud.yahoo.com> I'm looking for an HT position in either Madison, WI or Rockford, Ill or surronding area's. Thanks, Steve --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. From Joannah.Miley <@t> northwestpathology.com Wed Nov 28 12:31:17 2007 From: Joannah.Miley <@t> northwestpathology.com (Miley, Joannah C.) Date: Wed Nov 28 12:31:24 2007 Subject: [Histonet] Melan-A on Dako Immuno Stainer Message-ID: <719B46988560834BBE7D7F72EEB7C10A09C81BE8@HINET1.hinet.org> Our lab is demoing a Dako immuonstainer and I am having a hard time optimizing our Melan-A. The dilutions I have tried are 1:50, 1:100, and 1:200. I have tried antigen retreival with EDTA pH9 and Citrate buffer at pH 6. I am incubating in the primary and polymer for 20 minutes each. I am still having a lot of trouble with excessive muddy background. Any ideas?? Thanks, Joannah Miley, HT(ASCP) NW Pathology Bellingham, WA From SBaldwin <@t> compucyte.com Wed Nov 28 12:36:09 2007 From: SBaldwin <@t> compucyte.com (Scott Baldwin) Date: Wed Nov 28 12:36:17 2007 Subject: [Histonet] Upcoming Cold Spring Harbor Symposium Message-ID: To All Histonetters, On behalf of the Cold Spring Harbor Symposium, we would like to let everyone know about the following exciting upcoming symposium. For all that are interested, please note that the third day of the symposium is dedicated to Tissue Analysis and Histopathology applications. This is the first announcement of the Inaugural Quantitative Imaging Cytometry Symposium at the Cold Spring Harbor Laboratory, which has been established as part of the Quantitative Imaging Cytometry Regional Center of Excellence activities (http://www.compucyte.com/news-Cold%20Spring%20Release.htm). The symposium will take place at the Cold Spring Harbor, New York campus from January 30 to February 1, 2008. The meeting is structured to combine both the theory and practice of QIC, with morning seminars featuring a keynote lecturer and presentations by researchers employing the technology in their work. Confirmed presentations and speakers include: ? Time, Biochemistry, and Cell States: Role of cytometry in systems biology - James Jacobberger, Professor of Oncology, Case Comprehensive Cancer Center, Cleveland, OH ? Laser Scanning cytometry: Where it fits into modern biomedical analysis - William Telford, NIH/NCI, Bethesda, MD ? Application of Laser Scanning Cytometry to the Functional Analysis of a Protein Phosphatase in a Mouse Model of Parkinson's Disease - Nicholas Tonks, Director of Shared Resources, Cold Spring Harbor Laboratory Cancer Center, Cold Spring Harbor, NY ? Quantitative Imaging Cytometry: How it complements the analytical capabilities of flow cytometry and molecular biology techniques - Zbigniew Darzynkiewicz, Director, Brander Cancer Research Institute, Valhalla, NY ? Cell Fate Signaling in Tumor Cells - Shazib Pervaiz, Professor, National University of Singapore ? Potential Mechanisms for the Generation of Chromosome Aneuploidy in Human Cancer - John M. Lehman, Professor, Brody School of Medicine, East Carolina University, Greenville, NC ? The Role of Circulating Epithelial Tumor Cells (CETC) in the Metastatic Pathway - Katharina Pachmann, Professor, Department of Experimental Hematology and Oncology, Friedrich Schiller Universit?t, Jena, Germany ? Looking at Old Colors with New Lights: Applying quantitative imaging cytometry to histopathology - William Geddie, Department of Laboratory Medicine and Pathobiology, University of Toronto ? Quantitative Imaging Cytometry Applications in Pre-Clinical Drug Development - David Krull, GlaxoSmithKline Safety Assessment, Research Triangle Park, NC ? Quantitation of Caspase 3 Activation as a Pharmacodynamic Endpoint in Fine Needle Aspirate (FNA) Biopsies - Gloria Juan, Clinical Immunology, Amgen, Thousand Oaks, CA ? Quantitative Imaging of Biomarkers in Whole Sections and Tissue Microarrays - Viju Ananthanarayanan, University of Illinois at Chicago, IL Lectures are followed by small group laboratory sessions providing extensive hands-on practical opportunities for further skill development in applying quantitative imaging cytometry (solid phase laser scanning cytometry) techniques in cell- and tissue-based applications. These sessions will be guided by an expert-level faculty from universities and industry and will focus on the following areas: 1. Practical instruction in designing QIC experiments: Implementation of high-content cell- and tissue-based applications on the iCys? Imaging Cytometer, quantification of fluorescence and laser light loss, defining assay end-points, dye selection (fluorescent and chromatic), sample preparation for automated analysis, troubleshooting analytical and image performance. 2. Advanced data analysis techniques: Multiparameter cell cycle; DNA damage; high-content tissue microarray analysis. 3. Individualized application development assistance on requested applications will be offered by experienced staff. The Symposium represents an opportunity for users of iGeneration LSC technology to come together to develop advanced skills in the technology, exchange ideas with other researchers and users of LSC technology, and take advantage of customized application development services at no additional cost. Evening poster sessions will provide an opportunity for attendees to further share their work with other researchers. A full 3-day course fee of $1500 will cover admission to the morning symposia, all afternoon instruction and materials, a peer-reviewed poster session and reception, accommodations for three nights and meals for three days on the Cold Spring Harbor Laboratory campus. Partial registration packages are also available. Registration for the meeting is now open at www.imagingcytometrycenter.com, as is the Call for Poster Abstracts. Registration is limited to 75 people; early registration is therefore recommended. Scott Baldwin CompuCyte Corporation From jqb7 <@t> CDC.GOV Wed Nov 28 13:37:46 2007 From: jqb7 <@t> CDC.GOV (Bartlett, Jeanine (CDC/CCID/NCZVED)) Date: Wed Nov 28 13:37:59 2007 Subject: [Histonet] plastics Message-ID: <34BB307EFC9A65429BBB49E330675F72045E2260@LTA3VS003.ees.hhs.gov> Hello All! I would like to hear from those of you that use plastics for light histology. I'd like to know which tissue types you use it for, what type you use, and if it requires the use of a special knife, etc. Thanks in advance for your help! Jeanine Bartlett, BS, HT(ASCP)QIHC Centers for Disease Control and Prevention Infectious Diseases Pathology Branch 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov From settembr <@t> umdnj.edu Wed Nov 28 13:11:54 2007 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Wed Nov 28 15:58:21 2007 Subject: [Histonet] Melan-A on Dako Immuno Stainer Message-ID: Hello Joannah, I use Dako's Melan-A on FFPE human tissue @ 1:50 I use their TRS for 40 minutes in a steamer and incubate for 15 minutes room temp. I then use LSAB2, but I suspect that you'll get better results with your polymer or Dako's polymer Env+ Mouse or Flex. Good luck. Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ USA >>> "Miley, Joannah C." 11/28/07 1:31 PM >>> Our lab is demoing a Dako immuonstainer and I am having a hard time optimizing our Melan-A. The dilutions I have tried are 1:50, 1:100, and 1:200. I have tried antigen retreival with EDTA pH9 and Citrate buffer at pH 6. I am incubating in the primary and polymer for 20 minutes each. I am still having a lot of trouble with excessive muddy background. Any ideas?? Thanks, Joannah Miley, HT(ASCP) NW Pathology Bellingham, WA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From fong <@t> zoology.ubc.ca Wed Nov 28 17:07:21 2007 From: fong <@t> zoology.ubc.ca (Angelina Fong) Date: Wed Nov 28 17:08:17 2007 Subject: [Histonet] substance P receptor In-Reply-To: <000801c8319f$903347a0$839b2fcf@NEIL> References: <000801c8319f$903347a0$839b2fcf@NEIL> Message-ID: <474DF4A9.1040708@zoology.ubc.ca> Hi Neil, I have had some lovely substance P receptor (or Neurokinin 1 receptor) labelling in rat brains using a rabbit polyclonal antibody from Novus Biological/ I don't have the catalogue number on hand, but let me know if you want it. I have never processed tissue for NK1R labeling with picric acid, but have used just straight 4% PFA transcardial perfusion after saline and labelling/staining looks great, both in fluorescence or DAB. I have used a guinea pig anti-NK1R antibody that didn't work quite as well as the one from Novus. Let me know if you would like more specifics. Good luck Angelina > Does anybody have experience staining rat brain for the substance P receptor? Which company would you recommend? I have also read that many researchers fix tissue with paraformaldehyde and picric acid? Unfortunately, when we designed our experiments we did not anticipate that we would have to stain our tissue with this antibody and I was wondering how important is it to fix tissue with a paraformaldehyde/picric acid solution. Our rats were perfused with physiological saline followed by 4% paraformaldehyde. Brains were removed and postfixed for 72 hrs in the same solution, before being placed in 0.1% sodium azide/PB solution prior to sectioning on a Vibratome at 50 micron. > > Thanks for the help, > > Neil > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- ~~o~~o~~o~~o~~o~~o~~o~~o~~o~~o Angelina Y. Fong, Ph.D. Department of Zoology Biological Sciences Building 6270 University Boulevard University of British Columbia Vancouver, BC, V6T 1Z4 Canada Ph: (604) 822-5990 Fax: (604) 822-2416 Email: fong@zoology.ubc.ca From MVaughan4 <@t> ucok.edu Wed Nov 28 18:29:16 2007 From: MVaughan4 <@t> ucok.edu (MVaughan4@ucok.edu) Date: Wed Nov 28 18:29:49 2007 Subject: [Histonet] How about another cryostat request? In-Reply-To: Message-ID: Histonetters, I have read some of the archives about cryostats, some like Microm, others Leica. I finally managed to scrape together about 15K to spend on a cryostat, but the cheapest (not necessarily the best) I found was a Thermo Richard Allan HM550 SDM (Stripped-Down Model, haha) for 16K, but it recently went up to 20K so it is pretty much out of my market. I plan to use a cryostat to section mostly skin and artificial skin tissues about 4 um thick. Does anybody have any idea what I should be looking for? Thanks for your help. Mel Melville B. Vaughan, Ph. D. Assistant Professor of Biology Campus Coordinator, NSF Sure-Step program University of Central Oklahoma 100 N. University Drive Edmond, OK 73034 ----------------------------------------- **CONFIDENTIALITY** -This email (including any attachments) may contain confidential, proprietary and privileged information. Any unauthorized disclosure or use of this information is prohibited. From karling <@t> usc.edu Wed Nov 28 21:52:26 2007 From: karling <@t> usc.edu (Kar Ling) Date: Wed Nov 28 21:52:31 2007 Subject: [Histonet] rehydrating cryosections Message-ID: Hi, Is anyone out there do cryosectioning for snap-frozen muscle (without cryoprotection with sucrose)?? How do you rehydrate the cryosections before immunofluorescent staining? Thank you! Ling From kgreen <@t> hsh.org Thu Nov 29 05:20:47 2007 From: kgreen <@t> hsh.org (Green, Kathy) Date: Thu Nov 29 05:20:51 2007 Subject: [Histonet] question about coating blocks with paraffin for storage Message-ID: Dear Fellow Histotechs: We have always "dipped" out blocks after cutting them before we file them, so I'm inquiring if any of you do not "dip" or recoat with paraffin, have you had any problems with your blocks? Someone said the tissue would shrink & bugs would get into them??? Thanks for your help in this matter. Kathy Green, HT Manager Histology Laboratory Holy Spirit Hospital 503 N. 21st Street Camp Hill, PA 17011 kgreen@hsh.org (717) 763-2930 Blackberry: (717) 370-1726 Confidentiality Disclaimer: The information contained in this communication may be confidential, is intended for the use of the recipient named above, and may be legally privileged.If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication, or any of its contents, is strictly prohibited. If you received this communication in error, please resend this communication to the sender and delete the original message and any copy of it from your computer system. Thank You From Barry.R.Rittman <@t> uth.tmc.edu Thu Nov 29 06:24:10 2007 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Thu Nov 29 06:24:16 2007 Subject: [Histonet] question about coating blocks with paraffin for storage References: Message-ID: Kathy this is one of those it depends answers. Sealing the surface is essential, not only for blocks but for mounted section that are kept in reserve. A quick dip in paraffin wax while it will temporarily seal the surface doe not mean that the paraffin has fused with the block, it may have merely coated the surface. This can separate at a later stage especially with long term storage, temperature changes etc. It is, in my experience, preferable to run a hot blade over the surface to ensure that the surface is in fact sealed from the atmosphere. We have only had problems when storing tissue blocks for long periods of time when we have used the dipping method. Disadvantage of the hot blade is that it takes a little more time and you do lose a very small layer of tissue. However if you are going to recut some time in the future you will almost certainly lose some tissue anyway. Hope that this helps. Barry ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Green, Kathy Sent: Thu 11/29/2007 5:20 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] question about coating blocks with paraffin for storage Dear Fellow Histotechs: We have always "dipped" out blocks after cutting them before we file them, so I'm inquiring if any of you do not "dip" or recoat with paraffin, have you had any problems with your blocks? Someone said the tissue would shrink & bugs would get into them??? Thanks for your help in this matter. Kathy Green, HT Manager Histology Laboratory Holy Spirit Hospital 503 N. 21st Street Camp Hill, PA 17011 kgreen@hsh.org (717) 763-2930 Blackberry: (717) 370-1726 Confidentiality Disclaimer: The information contained in this communication may be confidential, is intended for the use of the recipient named above, and may be legally privileged.If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication, or any of its contents, is strictly prohibited. If you received this communication in error, please resend this communication to the sender and delete the original message and any copy of it from your computer system. Thank You _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From petepath <@t> yahoo.com Thu Nov 29 06:26:30 2007 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Thu Nov 29 06:26:38 2007 Subject: [Histonet] How about another cryostat request? Message-ID: <725281.5457.qm@web45113.mail.sp1.yahoo.com> You may want to have a look at the Leica 1500 which is used in many Mohs labs and I believe is close to your price range. It is an excellent low priced cryostat with many of the features of the more expensive 1850. These are distributed by Belair Instrument Co. in Springfield NJ., also a provider of excellent cryostat maintence. From lelmgren <@t> sunriselab.com Thu Nov 29 06:35:43 2007 From: lelmgren <@t> sunriselab.com (Laurie Elmgren) Date: Thu Nov 29 06:36:06 2007 Subject: [Histonet] IHC Auto Stainer Message-ID: <4A672C6AE0402D4A89ECE29E8A4B47E32B74FC@MailPDC.sunriselab.com> We are exploring the options of doing IHC in house. There is so much press about the Ventanna XT. Does anyone have any input or experience with the Leica Bond Max Immunostainer? Laurie Elmgren Histology Supervisor Sunrise Medical Labs From Ronald.Houston <@t> nationwidechildrens.org Thu Nov 29 07:22:03 2007 From: Ronald.Houston <@t> nationwidechildrens.org (Houston, Ronald) Date: Thu Nov 29 07:22:34 2007 Subject: [Histonet] IHC Auto Stainer In-Reply-To: <4A672C6AE0402D4A89ECE29E8A4B47E32B74FC@MailPDC.sunriselab.com> Message-ID: <979FF5962E234F45B06CF0DB7C1AABB213A41643@chi2k3ms01.columbuschildrens.net> It's wonderful; to try to compare the two is like comparing oil and water Ronnie Houston, MS, HT(ASCP)QIHC Anatomic Pathology Manager Nationwide Children's Hospital -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie Elmgren Sent: Thursday, November 29, 2007 7:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC Auto Stainer We are exploring the options of doing IHC in house. There is so much press about the Ventanna XT. Does anyone have any input or experience with the Leica Bond Max Immunostainer? Laurie Elmgren Histology Supervisor Sunrise Medical Labs _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mauger <@t> email.chop.edu Thu Nov 29 07:39:15 2007 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Thu Nov 29 07:38:46 2007 Subject: [Histonet] IHC Auto Stainer Message-ID: Laurie, I am very impressed with the Leica Bondmax. Their detection is beautiful-it blows everyone else away. Get a demo and check it out. It also does ISH totally on board, and the results are great. Jo From mike.collins <@t> imperial.ac.uk Thu Nov 29 07:46:59 2007 From: mike.collins <@t> imperial.ac.uk (Collins, Michael) Date: Thu Nov 29 07:47:24 2007 Subject: [Histonet] Substance P Message-ID: Hi Neil! I recently used SP on buffered formalin perfused rat and guinea pig. Peninsula T-4106 1 : 250 in PBS + 0.1% BSA + 0.025% Tween 20 60 mins. Swine anti rabbit 1: 150 in that PBS 60 mins. ABC in normal PBS 30 mins. Aliquotted DAB 50 secs. Mike. From JeromeS <@t> rice.edu Thu Nov 29 08:28:26 2007 From: JeromeS <@t> rice.edu (J. Saltarrelli) Date: Thu Nov 29 08:28:34 2007 Subject: [Histonet] Immunocytochemistry question Message-ID: Hello all, I have an ICC dilemma and was hoping that someone would be able to help. I'm using ICC to test the phenotype of endothelial cells using anti-von Willebrand factor. The primary was produced in rabbit. To try and alleviate non-specific binding, as a control I used an isotype IgG that is also produced in rabbit. The secondary, FITC, is against rabbit. Could using an isotype IgG for blocking from the same species as the primary lead to excessive non-specific binding?? Would running the test using IgG from rat possibly reduce non-specific binding since the secondary is anti-rabbit?? Thanks for all of your help Thank you, J. Saltarrelli, Ph.D. Postdoctoral Research Fellow Department of Bioengineering Rice University From tkngflght <@t> yahoo.com Thu Nov 29 08:55:06 2007 From: tkngflght <@t> yahoo.com (Cheryl R. Kerry) Date: Thu Nov 29 08:55:39 2007 Subject: [Histonet] question about coating blocks with paraffin for storage In-Reply-To: Message-ID: <003001c83297$d5254860$6901a8c0@CHERYLSLAPTOP> Hi Kathy-- Y'know how we do things because 'we've always done them this way'? This is one of those situations and it's great that you're asking 'why'. Generally speaking--there is no need--even though with older processing and materials there USED to be. If your tissues are well processed using polymer waxes, you do not need to do this. They won't get bugs and there should be NO water, alcohol or hydrocarbons (xylene) left to dehydrate and shrink the tissues. With that said--if you have underprocessed tissues like uterus, fatty tissues or placenta that aren't completely fixed, dehydrated, cleared and infiltrated--you may have the centers of these tissues shrink back in a little bit. Having altered our procedures to save us this time every day, the best thing I can share it to do a control test! Leave a few days of blocks un-sealed and check them once a week for a month or two. It shouldn't take longer than that now that the heat is on in your facility. If there is no appreciable change--you're good to stop. If you see the larger tissues shrinking--then just seal those along the way. If all the tissues demonstrate some change--it's a sign they aren't completely processed and this should be addressed at processing, not storage (and this is unlikely as it would show on the slides.) Hope this helps--it may not seem like a huge time savings but if every tech takes 10 minutes a day--that is equal to almost one daily break--and who doesn't want to keep their breaks!!! Cheryl Cheryl R. Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab, one great tech at a time. 281.852.9457 office 281.883.7704 cell 800.756.3309 fax and alternate phone -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Green, Kathy Sent: Thursday, November 29, 2007 5:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] question about coating blocks with paraffin for storage Dear Fellow Histotechs: We have always "dipped" out blocks after cutting them before we file them, so I'm inquiring if any of you do not "dip" or recoat with paraffin, have you had any problems with your blocks? Someone said the tissue would shrink & bugs would get into them??? Thanks for your help in this matter. Kathy Green, HT Manager Histology Laboratory Holy Spirit Hospital 503 N. 21st Street Camp Hill, PA 17011 kgreen@hsh.org (717) 763-2930 Blackberry: (717) 370-1726 Confidentiality Disclaimer: The information contained in this communication may be confidential, is intended for the use of the recipient named above, and may be legally privileged.If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication, or any of its contents, is strictly prohibited. If you received this communication in error, please resend this communication to the sender and delete the original message and any copy of it from your computer system. Thank You _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TMcNemar <@t> lmhealth.org Thu Nov 29 09:02:33 2007 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Thu Nov 29 09:02:02 2007 Subject: [Histonet] Melan-A on Dako Immuno Stainer In-Reply-To: <719B46988560834BBE7D7F72EEB7C10A09C81BE8@HINET1.hinet.org> Message-ID: <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4FE@lmhsmail.lmhealth.org> We use Dako's Mel-A at 1:50, 20 min retrieval in a steamer using Dako's Target retrieval solution, and a 30 min incubation on the Autostainer. Works beautifully with the Envision + kit. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Miley, Joannah C. Sent: Wednesday, November 28, 2007 1:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Melan-A on Dako Immuno Stainer Our lab is demoing a Dako immuonstainer and I am having a hard time optimizing our Melan-A. The dilutions I have tried are 1:50, 1:100, and 1:200. I have tried antigen retreival with EDTA pH9 and Citrate buffer at pH 6. I am incubating in the primary and polymer for 20 minutes each. I am still having a lot of trouble with excessive muddy background. Any ideas?? Thanks, Joannah Miley, HT(ASCP) NW Pathology Bellingham, WA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Thu Nov 29 09:34:55 2007 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Thu Nov 29 09:35:04 2007 Subject: AW: [Histonet] question about coating blocks with paraffin for storage In-Reply-To: Message-ID: <001101c8329d$664ac6d0$6412a8c0@dielangs.at> We never sealed the blocks and never had any problems. One thing is, that in the storage boxes, the blocks sit so tight, that they probably seal one another. Gudrun Lang Biomed. Analytikerin Histolabor Akh Linz Krankenhausstr. 9 4020 Linz +43(0)732/7806-6754 -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Green, Kathy Gesendet: Donnerstag, 29. November 2007 12:21 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] question about coating blocks with paraffin for storage Dear Fellow Histotechs: We have always "dipped" out blocks after cutting them before we file them, so I'm inquiring if any of you do not "dip" or recoat with paraffin, have you had any problems with your blocks? Someone said the tissue would shrink & bugs would get into them??? Thanks for your help in this matter. Kathy Green, HT Manager Histology Laboratory Holy Spirit Hospital 503 N. 21st Street Camp Hill, PA 17011 kgreen@hsh.org (717) 763-2930 Blackberry: (717) 370-1726 Confidentiality Disclaimer: The information contained in this communication may be confidential, is intended for the use of the recipient named above, and may be legally privileged.If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication, or any of its contents, is strictly prohibited. If you received this communication in error, please resend this communication to the sender and delete the original message and any copy of it from your computer system. Thank You _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cmiller <@t> physlab.com Thu Nov 29 09:42:37 2007 From: cmiller <@t> physlab.com (Cheri Miller) Date: Thu Nov 29 09:42:44 2007 Subject: [Histonet] question about coating blocks with paraffin for storage In-Reply-To: <001101c8329d$664ac6d0$6412a8c0@dielangs.at> References: <001101c8329d$664ac6d0$6412a8c0@dielangs.at> Message-ID: <012001c8329e$77fe8690$3402a8c0@plab.local> We seal the stinky cysts! Cheri Miller HT ASCP Histology Supervisor Physicians Laboratory Services, Inc. Omaha, NE 68117 402 738 5052 PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Thursday, November 29, 2007 9:35 AM To: 'Green, Kathy' Cc: histonet@lists.utsouthwestern.edu Subject: AW: [Histonet] question about coating blocks with paraffin for storage We never sealed the blocks and never had any problems. One thing is, that in the storage boxes, the blocks sit so tight, that they probably seal one another. Gudrun Lang Biomed. Analytikerin Histolabor Akh Linz Krankenhausstr. 9 4020 Linz +43(0)732/7806-6754 -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Green, Kathy Gesendet: Donnerstag, 29. November 2007 12:21 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] question about coating blocks with paraffin for storage Dear Fellow Histotechs: We have always "dipped" out blocks after cutting them before we file them, so I'm inquiring if any of you do not "dip" or recoat with paraffin, have you had any problems with your blocks? Someone said the tissue would shrink & bugs would get into them??? Thanks for your help in this matter. Kathy Green, HT Manager Histology Laboratory Holy Spirit Hospital 503 N. 21st Street Camp Hill, PA 17011 kgreen@hsh.org (717) 763-2930 Blackberry: (717) 370-1726 Confidentiality Disclaimer: The information contained in this communication may be confidential, is intended for the use of the recipient named above, and may be legally privileged.If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication, or any of its contents, is strictly prohibited. If you received this communication in error, please resend this communication to the sender and delete the original message and any copy of it from your computer system. Thank You _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. From rjbuesa <@t> yahoo.com Thu Nov 29 10:36:37 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 29 10:36:44 2007 Subject: [Histonet] Melan-A on Dako Immuno Stainer In-Reply-To: <719B46988560834BBE7D7F72EEB7C10A09C81BE8@HINET1.hinet.org> Message-ID: <801746.53957.qm@web61222.mail.yahoo.com> With the DAKO autostainer I used Melan-A at 1:50 x 30 min after HIER with citrate at pH6 with good results. Ren? J. "Miley, Joannah C." wrote: Our lab is demoing a Dako immuonstainer and I am having a hard time optimizing our Melan-A. The dilutions I have tried are 1:50, 1:100, and 1:200. I have tried antigen retreival with EDTA pH9 and Citrate buffer at pH 6. I am incubating in the primary and polymer for 20 minutes each. I am still having a lot of trouble with excessive muddy background. Any ideas?? Thanks, Joannah Miley, HT(ASCP) NW Pathology Bellingham, WA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. From dmccaig <@t> ckha.on.ca Thu Nov 29 10:14:16 2007 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Thu Nov 29 10:45:58 2007 Subject: [Histonet] ORIENTATION OF ENDOSCOPIC BIOPSIES Message-ID: Hi Does anyone have any suggetions (other than eosin in the processor) to assist in have the endoscopic biopsies orientated on the proper plane when embedded. I feel we have too high of a percentage that are not getting embedded properly. diana From HornHV <@t> archildrens.org Thu Nov 29 10:49:25 2007 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Thu Nov 29 10:49:54 2007 Subject: [Histonet] semantics? Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6317D82A04@EMAIL.archildrens.org> Is it a reticular stain or a reticulum stain? On our labels for the retic stain we put the word, reticulum. One of my pathologists has asked me to correct this with the word, reticular. So, for 35 years have I been using the wrong word? Yikes. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3912 visit us on the web at: www.archildrens.org From rjbuesa <@t> yahoo.com Thu Nov 29 10:50:09 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 29 10:50:13 2007 Subject: [Histonet] IHC Auto Stainer In-Reply-To: <4A672C6AE0402D4A89ECE29E8A4B47E32B74FC@MailPDC.sunriselab.com> Message-ID: <174391.30539.qm@web61219.mail.yahoo.com> Leica Bond Max beats Ventana hands down! Ren? J. Laurie Elmgren wrote: We are exploring the options of doing IHC in house. There is so much press about the Ventanna XT. Does anyone have any input or experience with the Leica Bond Max Immunostainer? Laurie Elmgren Histology Supervisor Sunrise Medical Labs _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. From JWEEMS <@t> sjha.org Thu Nov 29 10:51:59 2007 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Thu Nov 29 10:52:15 2007 Subject: [Histonet] semantics? In-Reply-To: <9AE8AA9E1F644B4AA6C155FB6FD51C6317D82A04@EMAIL.archildrens.org> References: <9AE8AA9E1F644B4AA6C155FB6FD51C6317D82A04@EMAIL.archildrens.org> Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD3204CF93E0@sjhaexc02.sjha.org> Or then, it could be reticulin - which is preferred here... -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Thursday, November 29, 2007 11:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] semantics? Is it a reticular stain or a reticulum stain? On our labels for the retic stain we put the word, reticulum. One of my pathologists has asked me to correct this with the word, reticular. So, for 35 years have I been using the wrong word? Yikes. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3912 visit us on the web at: www.archildrens.org _______________________________________________ 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 Joseph's Health System, Inc. From rjbuesa <@t> yahoo.com Thu Nov 29 10:54:34 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 29 10:54:38 2007 Subject: [Histonet] semantics? In-Reply-To: <1CD6831EB9B26D45B0A3EAA79F7EBD3204CF93E0@sjhaexc02.sjha.org> Message-ID: <952396.50913.qm@web61211.mail.yahoo.com> Reticular is a qualifier, reticulin a name. You stain for reticulin, even if the end product has a reticular appearance. Ren? J. "Weems, Joyce" wrote: Or then, it could be reticulin - which is preferred here... -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Thursday, November 29, 2007 11:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] semantics? Is it a reticular stain or a reticulum stain? On our labels for the retic stain we put the word, reticulum. One of my pathologists has asked me to correct this with the word, reticular. So, for 35 years have I been using the wrong word? Yikes. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3912 visit us on the web at: www.archildrens.org _______________________________________________ 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 Joseph's Health System, Inc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. From rjbuesa <@t> yahoo.com Thu Nov 29 10:56:00 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 29 10:56:09 2007 Subject: [Histonet] semantics? In-Reply-To: <9AE8AA9E1F644B4AA6C155FB6FD51C6317D82A04@EMAIL.archildrens.org> Message-ID: <113877.70492.qm@web61214.mail.yahoo.com> No, you have not been wrong. Find out your pathologist's birthday and present him with a Webster desk edition. Ren? J. "Horn, Hazel V" wrote: Is it a reticular stain or a reticulum stain? On our labels for the retic stain we put the word, reticulum. One of my pathologists has asked me to correct this with the word, reticular. So, for 35 years have I been using the wrong word? Yikes. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3912 visit us on the web at: www.archildrens.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Never miss a thing. Make Yahoo your homepage. From TJJ <@t> Stowers-Institute.org Thu Nov 29 11:39:32 2007 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Thu Nov 29 11:39:48 2007 Subject: [Histonet] Re: Immunocytochemistry question Message-ID: Jerome, you wrote: >>I'm using ICC to test the phenotype of endothelial cells using anti-von Willebrand factor. The primary was produced in rabbit. To try and alleviate non-specific binding, as a control I used an isotype IgG that is also produced in rabbit. The secondary, FITC, is against rabbit. Could using an isotype IgG for blocking from the same species as the primary lead to excessive non-specific binding?? Would running the test using IgG from rat possibly reduce non-specific binding since the secondary is anti-rabbit??<< I think you're confusing where to use the isotype IgG. That particular reagent should be used as a negative control on a separate slide, using the same Ig concentration as your primary antibody (in ug/ml). If you use this as a serum block prior to antibody incubation (which is what I think you're asking), your anti-rabbit secondary antibody will recognize all the sites the rabbit IgG is located and bind everywhere. You will want to block with a 5-10% solution of normal serum of the animal your secondary antibody was made in. Your FITC labeled anti-rabbit antibody was made in what host species? Goat? Donkey? Swine? Whatever the answer is, you'll want to use that species' normal serum as a block prior to incubation in primary antibody. You can also use it as an antibody diluent for the primary and secondary antibodies if you wish. Good luck! Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 From LINDA.MARGRAF <@t> childrens.com Thu Nov 29 11:49:33 2007 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Thu Nov 29 11:49:58 2007 Subject: [Histonet] Position for Senior scientist Message-ID: <474EA74D020000DA00019275@CNET3.CHILDRENS.COM> Here's a message I was asked to post (please send inquiries to them not me). Thanks Linda M Histonet administrator ............ Senior Scientist / Manager Carpinteria, CA A position as Senior Scientist / Manager is open in R&D at Dako North America. The new person will head our Special Stains development group. Dako currently has 23 Special Stains products in a growing portfolio that will be expanded to match all staining needs in America and Europe. Some travel activity is expected to visit external contacts and for conferences. Good interpersonal skills will be needed for staff relations, in-house team collaboration and external contacts. The new person will be familiar with assay / product development, testing / validation and or troubleshooting. New stains projects develop, transfer and launch waves of 4 new products at a time. Additionally the group supports product maintenance including optimization of existing products to better meet current market demands. We have a network of external collaborators and advisors plus outsourcing partners that all facilitate our work and assure the capacity for continuous high product development quality. We are working according to ISO and FDA standards and internal Design Control procedures. We have a relaxed business work style with 40 hours flexible week schedules and have project teams that include members from other departments and or external collaborators. Requirements. Hospital Laboratory or Pathology Laboratory experience Experience with assay development and or troubleshooting Experience with Special Stains preferred, plus training / experience in one or more of Histology, Pathology and Oncology with additional Immunohistochemistry (Immunology) beneficial. Team working skills, including constructive utilization of diverse skills Presentation of own work in written and or oral format Constructive interaction with external experts, collaborators, partners and customers Adeptly following deadlines (or work towards updating these) Dako is an international, biomedical products company that manufactures and markets instrumentation, reagents and diagnostic kits for use in clinical and research laboratories. Dako is one of the world's leading companies in cancer diagnostics. We are devoted to helping pathologists by improving their ability to diagnose cancer. Dako is a global business with headquarters in Denmark. It was founded by the Danish medical doctor Niels Harboe in 1966 and has 40 years' experience in the development and production of reagents and antibodies. Dako provides excellent benefits including health, dental, vision, prescription card, 401k with Dako match and a competitive salary. No phone calls or 3rd parties please. Applications will only be accepted for current open positions. Dako is an Equal Employment Employer. Women and minority candidates are encouraged to apply. Please email or fax your resume, (word attachments only), with salary history to 805-684-5935 or hr.ca@dako.com ------------------------ From TMcNemar <@t> lmhealth.org Thu Nov 29 12:01:09 2007 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Thu Nov 29 12:00:43 2007 Subject: [Histonet] semantics? In-Reply-To: <9AE8AA9E1F644B4AA6C155FB6FD51C6317D82A04@EMAIL.archildrens.org> Message-ID: <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4FF@lmhsmail.lmhealth.org> Reticulin here. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Horn, Hazel V Sent: Thursday, November 29, 2007 11:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] semantics? Is it a reticular stain or a reticulum stain? On our labels for the retic stain we put the word, reticulum. One of my pathologists has asked me to correct this with the word, reticular. So, for 35 years have I been using the wrong word? Yikes. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3912 visit us on the web at: www.archildrens.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ryaskovich <@t> dir.nidcr.nih.gov Thu Nov 29 12:10:12 2007 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR) [E]) Date: Thu Nov 29 12:10:18 2007 Subject: [Histonet] semantics? In-Reply-To: <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4FF@lmhsmail.lmhealth.org> References: <9AE8AA9E1F644B4AA6C155FB6FD51C6317D82A04@EMAIL.archildrens.org> <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4FF@lmhsmail.lmhealth.org> Message-ID: A.F.I.P. says Reticulum. Ruth Yaskovich N.I.H. -----Original Message----- From: Tom McNemar [mailto:TMcNemar@lmhealth.org] Sent: Thursday, November 29, 2007 1:01 PM To: Horn, Hazel V; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] semantics? Reticulin here. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Horn, Hazel V Sent: Thursday, November 29, 2007 11:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] semantics? Is it a reticular stain or a reticulum stain? On our labels for the retic stain we put the word, reticulum. One of my pathologists has asked me to correct this with the word, reticular. So, for 35 years have I been using the wrong word? Yikes. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3912 visit us on the web at: www.archildrens.org _______________________________________________ Histonet mailing list Histonet@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 Shirley_PHUA <@t> hsa.gov.sg Thu Nov 29 12:06:04 2007 From: Shirley_PHUA <@t> hsa.gov.sg (Shirley PHUA) Date: Thu Nov 29 12:12:49 2007 Subject: [Histonet] Shirley Phua is out-of-office on 29 November 2007 afternoon. Message-ID: I will be out of the office from 29-11-2007 to 30-11-2007. I'll be out-of-office on 29 November 2007. I'll be back on 30 November 2007. Pathologists: I will process your requests when I return. If urgent, please forward your email to Henry_Kyaw@hsa.gov.sg From RSRICHMOND <@t> aol.com Thu Nov 29 12:19:49 2007 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Thu Nov 29 12:19:55 2007 Subject: [Histonet] Re: reticulum stain (was semantics?) Message-ID: The preferred name for this venerable silver technique is "reticulum stain". Whatever reticulin is (or was), it isn't all that the technique demonstrates. I never heard anybody say "reticular stain". How are people handling the hazmats problems with reticwhatever stains? Is anybody still using uranyl nitrate? Bob Richmond Samurai Pathologist Knoxville TN From JWEEMS <@t> sjha.org Thu Nov 29 12:21:24 2007 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Thu Nov 29 12:21:56 2007 Subject: [Histonet] Re: reticulum stain (was semantics?) In-Reply-To: Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD32048F4361@sjhaexc02.sjha.org> We are and put in the haul away waste... Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Robert Richmond Sent: Thursday, November 29, 2007 1:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: reticulum stain (was semantics?) The preferred name for this venerable silver technique is "reticulum stain". Whatever reticulin is (or was), it isn't all that the technique demonstrates. I never heard anybody say "reticular stain". How are people handling the hazmats problems with reticwhatever stains? Is anybody still using uranyl nitrate? Bob Richmond Samurai Pathologist Knoxville TN _______________________________________________ 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 Joseph's Health System, Inc. From RSRICHMOND <@t> aol.com Thu Nov 29 12:22:10 2007 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Thu Nov 29 12:22:17 2007 Subject: [Histonet] Re: question about coating blocks with paraffin for storage Message-ID: The late great R.D. Lillie, who spent his later years at LSU, pointed out that in the warm wet climate of Louisiana if you didn't seal your paraffin blocks the cockroaches would eat the tissue out of them. I've never seen this, but I've never practiced in Louisiana either. Bob Richmond Samurai Pathologist Knoxville TN From nbhatia <@t> dermatology.wisc.edu Thu Nov 29 12:28:58 2007 From: nbhatia <@t> dermatology.wisc.edu (Neehar Bhatia) Date: Thu Nov 29 12:29:10 2007 Subject: [Histonet] Immunofluorescence Message-ID: <20071129122858056.00000002868@derm-HF55T81> Generator Microsoft Word 11 (filtered medium) Hi Everybody, I plan to do immunofluorescence in mouse tissue. I saw a protocol where they have fixed the tissue in 95/1% ethanol/acetic acid and the made paraffin blocks. Why is this fixation protocol used instead of formalin? Does it help in eliminating autofluorescence? Thanks for your help Neehar Bhatia, Ph.D Assistant Scientist, Department of Dermatology, University of Wisconsin, 445 Henry Mall, Rm 302, Madison WI 53706 Ph 608.262.3239 Fax 608.263.2919 From wstover <@t> vet.uga.edu Thu Nov 29 12:29:38 2007 From: wstover <@t> vet.uga.edu (Wanda Stover) Date: Thu Nov 29 12:29:43 2007 Subject: [Histonet] UGA Workshop Message-ID: There will be a immunohistochemistry workshop at the University of Georgia's College of Vet Med on Jan. 12, 2008. This course will provide histologists and lab techs with the essential information needed to understand the theory and practice of immunohistochemistry. At completion, the histologist will be familiar with the appropriate terminology, basic theory and techniques, development of new assays, and troubleshooting. This course will provide 7.5 C.E. hours. Please contact me for the Agenda and or registration form. 706 542-5835 Wanda Stover Cost is: $100.00 for students Non students $125.00 Wanda Stover UGA VET MED Histology Athens, Ga. 30602 706 583-0474 From oshel1pe <@t> cmich.edu Thu Nov 29 12:34:04 2007 From: oshel1pe <@t> cmich.edu (Philip Oshel) Date: Thu Nov 29 12:34:19 2007 Subject: [Histonet] Uranyl nitrate disposal In-Reply-To: References: Message-ID: Uranyl nitrate and Uranyl acetate are routinely used as stains for transmission electron microscopy (really contrasting agents, not stains in the light microscopy sense). Both for sections and negative stains of bacteria, viruses, and macromolecules. Waste disposal is just as a toxic compound (uranium is toxic) in water. The radioactivity is too low to bother about, even for the safety people -- although there are probably some safety offices that regulate UNO3 and UAc they same way they do carbon-14 or phosphorus-32, which are issues. Phil >The preferred name for this venerable silver technique is "reticulum >stain". Whatever reticulin is (or was), it isn't all that the >technique demonstrates. I never heard anybody say "reticular stain". > >How are people handling the hazmats problems with reticwhatever >stains? Is anybody still using uranyl nitrate? > >Bob Richmond >Samurai Pathologist >Knoxville TN > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Microscopy Facility Supervisor Biology Department 024C Brooks Hall Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 From rjbuesa <@t> yahoo.com Thu Nov 29 12:36:58 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Nov 29 12:37:02 2007 Subject: [Histonet] Re: question about coating blocks with paraffin for storage In-Reply-To: Message-ID: <532382.97773.qm@web61222.mail.yahoo.com> But I did in Cuba, and there were also "FFPE" tissue eating cockroaches (although perhaps not now, they probably have died of lack of food!). Ren? J. Robert Richmond wrote: The late great R.D. Lillie, who spent his later years at LSU, pointed out that in the warm wet climate of Louisiana if you didn't seal your paraffin blocks the cockroaches would eat the tissue out of them. I've never seen this, but I've never practiced in Louisiana either. Bob Richmond Samurai Pathologist Knoxville TN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. From Barry.R.Rittman <@t> uth.tmc.edu Thu Nov 29 12:44:58 2007 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Thu Nov 29 12:45:03 2007 Subject: [Histonet] Re: question about coating blocks with paraffin forstorage In-Reply-To: <532382.97773.qm@web61222.mail.yahoo.com> Message-ID: It is not only insects but also the possibility of various fungi invading the tissue. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Thursday, November 29, 2007 12:37 PM To: Robert Richmond; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Re: question about coating blocks with paraffin forstorage But I did in Cuba, and there were also "FFPE" tissue eating cockroaches (although perhaps not now, they probably have died of lack of food!). Ren? J. Robert Richmond wrote: The late great R.D. Lillie, who spent his later years at LSU, pointed out that in the warm wet climate of Louisiana if you didn't seal your paraffin blocks the cockroaches would eat the tissue out of them. I've never seen this, but I've never practiced in Louisiana either. Bob Richmond Samurai Pathologist Knoxville TN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From b-frederick <@t> northwestern.edu Thu Nov 29 13:01:15 2007 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Thu Nov 29 13:01:28 2007 Subject: [Histonet] semantics? In-Reply-To: <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4FF@lmhsmail.lmhealth.org> Message-ID: <000001c832ba$3ad591c0$d00f7ca5@lurie.northwestern.edu> We just say Retic. No semantic issues at all. Now as to which method..... Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tom McNemar Sent: Thursday, November 29, 2007 12:01 PM To: Horn, Hazel V; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] semantics? Reticulin here. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Horn, Hazel V Sent: Thursday, November 29, 2007 11:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] semantics? Is it a reticular stain or a reticulum stain? On our labels for the retic stain we put the word, reticulum. One of my pathologists has asked me to correct this with the word, reticular. So, for 35 years have I been using the wrong word? Yikes. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3912 visit us on the web at: www.archildrens.org _______________________________________________ Histonet mailing list Histonet@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 cornettl <@t> hotmail.com Thu Nov 29 13:10:24 2007 From: cornettl <@t> hotmail.com (Lorraine Cornett) Date: Thu Nov 29 13:10:33 2007 Subject: [Histonet] Amyloid Control Message-ID: I am appealing to the histonet to help me out. We have totally exhausted all amyloid control blocks that we can get our hands on. My pathologists don't want to order control slides, they want a block that can be cut when the stain is ordered. Does anyone out there have extra amyloid control blocks that they could share? Thanks in advance for any help!Lorraine Cornett, HT (ASCP)Highlands PathologyBlue Ridge Division, Kingsport, TN423 224-5793 fax 423 224-5349 _________________________________________________________________ Connect and share in new ways with Windows Live. http://www.windowslive.com/connect.html?ocid=TXT_TAGLM_Wave2_newways_112007 From failm <@t> musc.edu Thu Nov 29 13:09:19 2007 From: failm <@t> musc.edu (Fail, Mildred M.) Date: Thu Nov 29 13:10:44 2007 Subject: [Histonet] Retiring 2/1/08 Message-ID: I am retiring February 1st, If you are looking for a great place to live and work try Charleston, SC. Wonderful beaches, beautiful gardens.Spoleto in the spring. The job is challenging, but there is so much variety, boredom is never a problem. Immunohistochemistry, immunofluorescence, muscle histochemistry, special stains, And still so much to learn. My boss is always open to learning new techniques.And being the editor of the Histologic encourages the staff to share their expertise with others by submitting articles to the Histologic. The salary being offered is quite good. (A bit more than I'm paid actually)I've been fortunate, to be able to work at a profession that I enjoyed in a institution I respected. But its time for me to spend some time with family and play at other hobbies. From dennijc <@t> vetmed.auburn.edu Thu Nov 29 13:14:55 2007 From: dennijc <@t> vetmed.auburn.edu (John C. Dennis) Date: Thu Nov 29 13:15:21 2007 Subject: [Histonet] Immunocytochemistry question In-Reply-To: References: Message-ID: Dear J Saltarrelli Go ahead and do the experiment. Run your protocol without primary and see what sort of non-specific signal there is to be seen. Or does that open the empiricism vs rationalism can of worms? John Carroll Dennis Anatomy, Physiology, and Pharmacology 109 Greene Hall Auburn University, AL 36849 On Thu, 29 Nov 2007, J. Saltarrelli wrote: > Hello all, > > > > I have an ICC dilemma and was hoping that someone would be able to help. > > > > I'm using ICC to test the phenotype of endothelial cells using anti-von > Willebrand factor. The primary was produced in rabbit. To try and > alleviate non-specific binding, as a control I used an isotype IgG that is > also produced in rabbit. The secondary, FITC, is against rabbit. Could > using an isotype IgG for blocking from the same species as the primary lead > to excessive non-specific binding?? Would running the test using IgG from > rat possibly reduce non-specific binding since the secondary is > anti-rabbit?? > > > > Thanks for all of your help > > > > Thank you, > > > > J. Saltarrelli, Ph.D. > > Postdoctoral Research Fellow > > Department of Bioengineering > > Rice University > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gu.lang <@t> gmx.at Thu Nov 29 13:43:12 2007 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Thu Nov 29 13:43:18 2007 Subject: WG: [Histonet] semantics? Message-ID: <000401c832c0$13d0f4b0$6412a8c0@dielangs.at> "Gitterfaser-Faerbung" ;) Gudrun Lang Biomed. Analytikerin Histolabor Akh Linz Krankenhausstr. 9 4020 Linz +43(0)732/7806-6754 -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Horn, Hazel V Gesendet: Donnerstag, 29. November 2007 17:49 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] semantics? Is it a reticular stain or a reticulum stain? On our labels for the retic stain we put the word, reticulum. One of my pathologists has asked me to correct this with the word, reticular. So, for 35 years have I been using the wrong word? Yikes. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3912 visit us on the web at: www.archildrens.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CIngles <@t> uwhealth.org Thu Nov 29 13:59:25 2007 From: CIngles <@t> uwhealth.org (Ingles Claire) Date: Thu Nov 29 14:02:59 2007 Subject: [Histonet] How about another cryostat request? References: Message-ID: <08A0A863637F1349BBFD83A96B27A50A1200A8@uwhis-xchng3.uwhis.hosp.wisc.edu> Mel: Have you checked around for reconditioned/used equipment? It is alot cheaper, and most companies still have warranties on the work. Hope this helps. Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of MVaughan4@ucok.edu Sent: Wed 11/28/2007 6:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] How about another cryostat request? Histonetters, I have read some of the archives about cryostats, some like Microm, others Leica. I finally managed to scrape together about 15K to spend on a cryostat, but the cheapest (not necessarily the best) I found was a Thermo Richard Allan HM550 SDM (Stripped-Down Model, haha) for 16K, but it recently went up to 20K so it is pretty much out of my market. I plan to use a cryostat to section mostly skin and artificial skin tissues about 4 um thick. Does anybody have any idea what I should be looking for? Thanks for your help. Mel Melville B. Vaughan, Ph. D. Assistant Professor of Biology Campus Coordinator, NSF Sure-Step program University of Central Oklahoma 100 N. University Drive Edmond, OK 73034 ----------------------------------------- **CONFIDENTIALITY** -This email (including any attachments) may contain confidential, proprietary and privileged information. Any unauthorized disclosure or use of this information is prohibited. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cad <@t> Stowers-Institute.org Thu Nov 29 14:11:39 2007 From: cad <@t> Stowers-Institute.org (Dickey, Coral) Date: Thu Nov 29 14:12:00 2007 Subject: [Histonet] Double skeletal stain in Zebrafish Message-ID: Hello, I am looking for a protocol for Alizarian red/Alcian blue double skeletal stain on whole zebrafish. I have a protocol for chick and mice embryos, but wanted something more specific to the species I am working with. Thanks, Coral Dickey Histology Specialist I Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, Missouri 64110 Phone: 816-926-4305 e-mail: cad@stowers-institute.org From mtarango <@t> nvcancer.org Thu Nov 29 15:57:20 2007 From: mtarango <@t> nvcancer.org (Tarango, Mark) Date: Thu Nov 29 15:58:29 2007 Subject: [Histonet] semantics? In-Reply-To: <51D5D78FBEDAEA4FBCCD9A9D44211DC528F4FF@lmhsmail.lmhealth.org> Message-ID: <5AEC610C1CE02945BD63A395BA763EDE011B6FED@NVCIEXCH02.NVCI.org> I think it's reticulum network, reticular fibers, or reticulin fibers that we're staining. I really don't think that the label needs to be corrected. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tom McNemar Sent: Thursday, November 29, 2007 10:01 AM To: Horn, Hazel V; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] semantics? Reticulin here. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Horn, Hazel V Sent: Thursday, November 29, 2007 11:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] semantics? Is it a reticular stain or a reticulum stain? On our labels for the retic stain we put the word, reticulum. One of my pathologists has asked me to correct this with the word, reticular. So, for 35 years have I been using the wrong word? Yikes. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3912 visit us on the web at: www.archildrens.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== From jerry.santiago <@t> jax.ufl.edu Thu Nov 29 18:44:36 2007 From: jerry.santiago <@t> jax.ufl.edu (Santiago, Jerry) Date: Thu Nov 29 18:44:56 2007 Subject: [Histonet] Florida Society for Histotechnology-Call for Abstracts Message-ID: <816DC61E1730E843A0BCF4CCFA1EFCF3065511@jaxmail.umc.ufl.edu> Call for Abstracts! Florida Society for Histotechnology is now accepting abstracts for their 2008 meeting. The deadline for abstracts is December 31st, 2007. The meeting if schedule for May 15-18, 2008 at the Bahia Mar Beach Resort in Fort Lauderdale. Abstracts may be sent via e-mail to fsh@fshgroup.org. Announcement of abstract acceptance will be announced at the end of January 2008. Exhibit information will be available as of December 3rd, 2007 on our website www.fshgroup.org From jnocito <@t> satx.rr.com Thu Nov 29 18:46:04 2007 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Thu Nov 29 18:46:08 2007 Subject: [Histonet] semantics? References: <5AEC610C1CE02945BD63A395BA763EDE011B6FED@NVCIEXCH02.NVCI.org> Message-ID: <019501c832ea$635acbd0$0302a8c0@yourxhtr8hvc4p> Scotch works for me Joe ----- Original Message ----- From: "Tarango, Mark" To: "Tom McNemar" ; "Horn, Hazel V" ; Sent: Thursday, November 29, 2007 3:57 PM Subject: RE: [Histonet] semantics? I think it's reticulum network, reticular fibers, or reticulin fibers that we're staining. I really don't think that the label needs to be corrected. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tom McNemar Sent: Thursday, November 29, 2007 10:01 AM To: Horn, Hazel V; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] semantics? Reticulin here. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Horn, Hazel V Sent: Thursday, November 29, 2007 11:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] semantics? Is it a reticular stain or a reticulum stain? On our labels for the retic stain we put the word, reticulum. One of my pathologists has asked me to correct this with the word, reticular. So, for 35 years have I been using the wrong word? Yikes. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3912 visit us on the web at: www.archildrens.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ============================================================================== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Susan.Walzer <@t> HCAHealthcare.com Fri Nov 30 02:40:45 2007 From: Susan.Walzer <@t> HCAHealthcare.com (Walzer Susan) Date: Fri Nov 30 02:42:39 2007 Subject: [Histonet] ORIENTATION OF ENDOSCOPIC BIOPSIES In-Reply-To: References: Message-ID: <471953BC63077941B82C26A4338272B42F0559@ORLEV03.hca.corpad.net> I was under the impression that most biopsies done with this procedure were like skin shave bx's, that is that they are disc shaped usually and need to be embedded on end in order to see all layers of mucosa. Often they come in all shapes and we do the best we can. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Diana McCaig Sent: Thursday, November 29, 2007 11:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] ORIENTATION OF ENDOSCOPIC BIOPSIES Hi Does anyone have any suggetions (other than eosin in the processor) to assist in have the endoscopic biopsies orientated on the proper plane when embedded. I feel we have too high of a percentage that are not getting embedded properly. diana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nyilmaz <@t> mersin.edu.tr Fri Nov 30 05:17:44 2007 From: nyilmaz <@t> mersin.edu.tr (Nejat) Date: Fri Nov 30 05:16:36 2007 Subject: [Histonet] mouse fetal brain processing Message-ID: <000501c83342$a15a31c0$5201a8c0@NEJAT1> Hi histonetters, Are there any suggestions on processing mouse fetal brains D15 and D17 except frozen sectioning? We had a lot of sectioning difficulties and artifacts with these FFPE tissues. Furthermore, any suggestions on preserving frozen tissues for re-sectioning for long periods? Dr. Necat Yilmaz Mersin Universitesi Tip Fakultesi Histoloji ve Embriyoloji Anabilim Dali From jqb7 <@t> CDC.GOV Fri Nov 30 05:16:58 2007 From: jqb7 <@t> CDC.GOV (Bartlett, Jeanine (CDC/CCID/NCZVED)) Date: Fri Nov 30 05:17:13 2007 Subject: [Histonet] Uranyl nitrate disposal In-Reply-To: References: Message-ID: <34BB307EFC9A65429BBB49E330675F72045E226A@LTA3VS003.ees.hhs.gov> We are not allowed to dispose of our uranyl nitrate in the normal hazardous waste. Our radiation group handles it. Jeanine Bartlett, BS, HT(ASCP)QIHC Centers for Disease Control and Prevention Infectious Diseases Pathology Branch 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Philip Oshel Sent: Thursday, November 29, 2007 1:34 PM To: Histonet@Pathology.swmed.edu Subject: [Histonet] Uranyl nitrate disposal Uranyl nitrate and Uranyl acetate are routinely used as stains for transmission electron microscopy (really contrasting agents, not stains in the light microscopy sense). Both for sections and negative stains of bacteria, viruses, and macromolecules. Waste disposal is just as a toxic compound (uranium is toxic) in water. The radioactivity is too low to bother about, even for the safety people -- although there are probably some safety offices that regulate UNO3 and UAc they same way they do carbon-14 or phosphorus-32, which are issues. Phil >The preferred name for this venerable silver technique is "reticulum >stain". Whatever reticulin is (or was), it isn't all that the technique >demonstrates. I never heard anybody say "reticular stain". > >How are people handling the hazmats problems with reticwhatever stains? >Is anybody still using uranyl nitrate? > >Bob Richmond >Samurai Pathologist >Knoxville TN > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Microscopy Facility Supervisor Biology Department 024C Brooks Hall Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMitchell <@t> uwhealth.org Fri Nov 30 09:37:22 2007 From: JMitchell <@t> uwhealth.org (Mitchell Jean A.) Date: Fri Nov 30 09:37:26 2007 Subject: [Histonet] Ultramicrotome Message-ID: <713A63AAF4280941A2EF88D66838F9A0FC40E4@uwhis-xchng4.uwhis.hosp.wisc.edu> I am looking to purchase a used ultramicrotome for electron microscopy. Would appreciate any suggestions as to where to begin hunting for one. Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Neuromuscular Laboratory Manager 600 Highland Avenue Madison, WI 53792-5132 From RBARNHART <@t> summithealth.org Fri Nov 30 09:53:24 2007 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Nov 30 09:54:06 2007 Subject: [Histonet] Slide quality Message-ID: <474FEBA4020000570000C7DF@carrierpigeon.SummitHealth.local> Recently we started monitoring slide quality in the histology department. We have noticed a trend of folds/wrinkles. I have changed paraffins from Surgipath Blue Ribbon to Fisher's Paraplast X-tra and it does not seem to have made a difference. I have tired using Halt which didn't make a difference either. Most of our specimens are GI biopsies, curettings, skins and breast biopsies. Any other suggestions? Thanks for the help. From rjbuesa <@t> yahoo.com Fri Nov 30 10:01:09 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 30 10:01:15 2007 Subject: [Histonet] Ultramicrotome In-Reply-To: <713A63AAF4280941A2EF88D66838F9A0FC40E4@uwhis-xchng4.uwhis.hosp.wisc.edu> Message-ID: <167138.53090.qm@web61215.mail.yahoo.com> Try eBay under microtomes. I have seen some there, or they lead to a used supplier. Ren? J. "Mitchell Jean A." wrote: I am looking to purchase a used ultramicrotome for electron microscopy. Would appreciate any suggestions as to where to begin hunting for one. Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Neuromuscular Laboratory Manager 600 Highland Avenue Madison, WI 53792-5132 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Never miss a thing. Make Yahoo your homepage. From rjbuesa <@t> yahoo.com Fri Nov 30 10:04:46 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 30 10:04:50 2007 Subject: [Histonet] Slide quality In-Reply-To: <474FEBA4020000570000C7DF@carrierpigeon.SummitHealth.local> Message-ID: <479329.35997.qm@web61223.mail.yahoo.com> Usually wrinkles are the consequence of poor infiltration or poor microtomy practices more than on the paraffin's quality. Check if you have changed anything on your processing schedule or in some of the techs' sectioning habits. Do wrinkles appear on sections from everybody? A section QA program has to be able to identify who prepared the slides. Ren? J. Rebecca Barnhart wrote: Recently we started monitoring slide quality in the histology department. We have noticed a trend of folds/wrinkles. I have changed paraffins from Surgipath Blue Ribbon to Fisher's Paraplast X-tra and it does not seem to have made a difference. I have tired using Halt which didn't make a difference either. Most of our specimens are GI biopsies, curettings, skins and breast biopsies. Any other suggestions? Thanks for the help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. From doug <@t> ppspath.com Fri Nov 30 10:18:21 2007 From: doug <@t> ppspath.com (Douglas D Deltour) Date: Fri Nov 30 10:18:39 2007 Subject: SPAM-LOW: [Histonet] Slide quality In-Reply-To: <474FEBA4020000570000C7DF@carrierpigeon.SummitHealth.local> Message-ID: Rebecca, I would defiantly check to see if the wrinkles are generated by one particular person. My techs initial the slides that they cut. This is the easiest way to see if things of this nature are a trend. Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca Barnhart Sent: Friday, November 30, 2007 10:53 AM To: histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: [Histonet] Slide quality Recently we started monitoring slide quality in the histology department. We have noticed a trend of folds/wrinkles. I have changed paraffins from Surgipath Blue Ribbon to Fisher's Paraplast X-tra and it does not seem to have made a difference. I have tired using Halt which didn't make a difference either. Most of our specimens are GI biopsies, curettings, skins and breast biopsies. Any other suggestions? Thanks for the help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From staceylburton <@t> yahoo.com Fri Nov 30 10:31:25 2007 From: staceylburton <@t> yahoo.com (Stacey Burton) Date: Fri Nov 30 10:31:28 2007 Subject: [Histonet] remove from list Message-ID: <630300.59097.qm@web30007.mail.mud.yahoo.com> Please remove me from the mailing list. Thank you, Stacey Burton ____________________________________________________________________________________ Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. http://mobile.yahoo.com/sports;_ylt=At9_qDKvtAbMuh1G1SQtBI7ntAcJ From Lori.Disher <@t> HCAhealthcare.com Fri Nov 30 10:31:06 2007 From: Lori.Disher <@t> HCAhealthcare.com (Disher Lori) Date: Fri Nov 30 10:32:06 2007 Subject: [Histonet] Decal Solutions and Her2 Fish Message-ID: <095327C7CDBDF64B9E9728A54799091E33A593@ORLEV03.hca.corpad.net> Hello, Can anyone recomend a decal solution to use that will not destroy the cells for HER 2 Fish? If so, do you have a policy? Lori Disher Fawcett Memorial Hosp 21298 Olean Blvd Port Charlotte, FL 33952 941-627-6128 From ifc1028 <@t> yahoo.com Fri Nov 30 11:25:31 2007 From: ifc1028 <@t> yahoo.com (Irene Cochran) Date: Fri Nov 30 11:25:37 2007 Subject: [Histonet] sudan black stain for fecal fat Message-ID: <746750.38134.qm@web45608.mail.sp1.yahoo.com> Our pathologist is looking for a procedure on staining fecal fat with sudan black any suggestions? thanks --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. From jpiche-grocki <@t> wtbyhosp.org Fri Nov 30 11:33:53 2007 From: jpiche-grocki <@t> wtbyhosp.org (Piche-Grocki, Jessica) Date: Fri Nov 30 11:35:39 2007 Subject: [Histonet] CDX2 and HCA (hepatocellular carcinoma) Message-ID: <0A57D6AEAE4CBA4A984D27257160A72D01069D0B@win03exchange01.wtbyhosp.org> Hello, I am having a hard time finding information for our pathologists on CDX2 and HCA antibodies. If anyone has any information they can share that would be great. I am especially having a hard time with the HCA(hepatocellular carcinoma antigen). Thanks and have a great weekend!! Jessica Piche-Grocki, HT(ASCP) CONFIDENTIALITY NOTICE: This email and any attachments contain confidential information that is legally privileged. This information is intended only for the use of the individual or entity named above. The authorized recipient of this information is prohibited from disclosing this information to any other party unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution or action taken in reliance on the contents of these documents is strictly prohibited. If you have received this information in error, please notify the sender immediately and delete these documents. Copyright (c) Waterbury Hospital From rjbuesa <@t> yahoo.com Fri Nov 30 11:39:42 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 30 11:39:48 2007 Subject: [Histonet] Decal Solutions and Her2 Fish In-Reply-To: <095327C7CDBDF64B9E9728A54799091E33A593@ORLEV03.hca.corpad.net> Message-ID: <916852.26259.qm@web61213.mail.yahoo.com> We used RDO and EDTA for more delicate specimens (and BM) and after that we regularly did IHC on those sections. I don't think that Her2 neu FISH would behave any differently, although I never did that (FISH always on breast tumors not requiring decal). Ren? J. Disher Lori wrote: Hello, Can anyone recomend a decal solution to use that will not destroy the cells for HER 2 Fish? If so, do you have a policy? Lori Disher Fawcett Memorial Hosp 21298 Olean Blvd Port Charlotte, FL 33952 941-627-6128 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. From rjbuesa <@t> yahoo.com Fri Nov 30 11:41:57 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 30 11:42:03 2007 Subject: [Histonet] sudan black stain for fecal fat In-Reply-To: <746750.38134.qm@web45608.mail.sp1.yahoo.com> Message-ID: <57495.58444.qm@web61216.mail.yahoo.com> And don't you use Oil Red O? You have not processed the feces and could use the ORO Ren? J. Irene Cochran wrote: Our pathologist is looking for a procedure on staining fecal fat with sudan black any suggestions? thanks --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. From cornettl <@t> hotmail.com Fri Nov 30 11:49:26 2007 From: cornettl <@t> hotmail.com (Lorraine Cornett) Date: Fri Nov 30 11:49:31 2007 Subject: [Histonet] Amyloid Message-ID: Thanks to everyone on the histonet, I have a couple of control blocks on the way! Have a great weekend!Lorraine Cornett, HT (ASCP)Highlands PathologyBlue Ridge Division, Kingsport, TN423 224-5793 fax 423 224-5349 _________________________________________________________________ Your smile counts. The more smiles you share, the more we donate.? Join in. www.windowslive.com/smile?ocid=TXT_TAGLM_Wave2_oprsmilewlhmtagline From FMonson <@t> wcupa.edu Fri Nov 30 12:07:25 2007 From: FMonson <@t> wcupa.edu (Monson, Frederick ) Date: Fri Nov 30 12:07:35 2007 Subject: [Histonet] semantics? In-Reply-To: <019501c832ea$635acbd0$0302a8c0@yourxhtr8hvc4p> References: <5AEC610C1CE02945BD63A395BA763EDE011B6FED@NVCIEXCH02.NVCI.org> <019501c832ea$635acbd0$0302a8c0@yourxhtr8hvc4p> Message-ID: <641CEFFC7E5B6C42AB59539653FD082304E0B578@wcu-ex-emp2.PASSHE.LCL> I agree with Joe (here I am butting in again), but..... I like 'dissection' rather than 'disection' which terms some lexicographers like to treat as synonyms. Since I am write and they R rong, and I am in charge, all my minions have been dissectors rather than disectors. I always hope that my surgeon will be the former rather than the latter. Thus it goes with arbitrary, and even sometimes correct, bosses. Even when they are wrong, they assume the right to be left when they are talking to their techs. I assume that many pathologists have bottles of dye/stain on their shelves designated as either 'blue collagen' or 'pink collagen' stain which phrases they must also use in their presentations. In such cases their sub-ordinates merely have to kno the final color to choose the correct one. Of course a peer may ask at a luncheon, "What's the difference between red and pink collagen? Surely there must be a molecular explanation. Have you ever seen yellow collagen or green? Perhaps there is a paper here." I vote for the guy/gal who invented the stain. Surely s/he got it wrong for all of us to follow. Jeers, Cheers, and Beers, it's Friday! Fred Monson Frederick C. Monson, PhD Technical Director Microanalysis and Imaging Research and Training Center (MIRTC) Large Scientific Instrument Core Geology, West Chester University S. Church St. and W. Rosedale Ave. West Chester, PA, 19320 610-738-0437 fmonson@wcupa.edu New Scheduler: http://lexspiac.wcupa.edu/cgi-bin/ureserve_gold.pl Web Page: http://lexspiac.wcupa.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Thursday, November 29, 2007 7:46 PM To: Tarango, Mark; Tom McNemar; Horn, Hazel V; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] semantics? Scotch works for me Joe ----- Original Message ----- From: "Tarango, Mark" To: "Tom McNemar" ; "Horn, Hazel V" ; Sent: Thursday, November 29, 2007 3:57 PM Subject: RE: [Histonet] semantics? I think it's reticulum network, reticular fibers, or reticulin fibers that we're staining. I really don't think that the label needs to be corrected. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tom McNemar Sent: Thursday, November 29, 2007 10:01 AM To: Horn, Hazel V; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] semantics? Reticulin here. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Horn, Hazel V Sent: Thursday, November 29, 2007 11:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] semantics? Is it a reticular stain or a reticulum stain? On our labels for the retic stain we put the word, reticulum. One of my pathologists has asked me to correct this with the word, reticular. So, for 35 years have I been using the wrong word? Yikes. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3912 visit us on the web at: www.archildrens.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------ ------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ======================================================================== ====== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pierre.chaumat <@t> alphelys.com Fri Nov 30 12:08:04 2007 From: pierre.chaumat <@t> alphelys.com (Pierre CHAUMAT) Date: Fri Nov 30 12:08:17 2007 Subject: [Histonet] Wrinkles and folds in paraffin sections In-Reply-To: <0MKtd6-1IyA9w0OL4-0006re@mx.kundenserver.de> Message-ID: <0ML21M-1IyAHU2QEX-0002OL@mrelayeu.kundenserver.de> Dear Rebecca, To my understanding, wrinkles do usually appear when the speed of cutting is not correct with hesitations. The cutting has to be smooth, homogeneous speed without stops or hesitations. Too cold paraffin frequently increases this but it is usually mainly due to handling. Motorized microtome can also help by controlling speed but it costs ! Good luck Pierre Rebecca Barnhart wrote: Recently we started monitoring slide quality in the histology department. We have noticed a trend of folds/wrinkles. I have changed paraffins from Surgipath Blue Ribbon to Fisher's Paraplast X-tra and it does not seem to have made a difference. I have tired using Halt which didn't make a difference either. Most of our specimens are GI biopsies, curettings, skins and breast biopsies. Any other suggestions? Thanks for the help. -----Message d'origine----- De : histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] De la part de histonet-request@lists.utsouthwestern.edu Envoy? : vendredi 30 novembre 2007 19:00 ? : histonet@lists.utsouthwestern.edu Objet : Histonet Digest, Vol 48, Issue 42 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Re: Slide quality (Rene J Buesa) 2. RE: SPAM-LOW: [Histonet] Slide quality (Douglas D Deltour) 3. remove from list (Stacey Burton) 4. Decal Solutions and Her2 Fish (Disher Lori) 5. sudan black stain for fecal fat (Irene Cochran) 6. CDX2 and HCA (hepatocellular carcinoma) (Piche-Grocki, Jessica) 7. Re: Decal Solutions and Her2 Fish (Rene J Buesa) 8. Re: sudan black stain for fecal fat (Rene J Buesa) 9. Amyloid (Lorraine Cornett) ---------------------------------------------------------------------- Message: 1 Date: Fri, 30 Nov 2007 08:04:46 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Slide quality To: Rebecca Barnhart , histonet@lists.utsouthwestern.edu Message-ID: <479329.35997.qm@web61223.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Usually wrinkles are the consequence of poor infiltration or poor microtomy practices more than on the paraffin's quality. Check if you have changed anything on your processing schedule or in some of the techs' sectioning habits. Do wrinkles appear on sections from everybody? A section QA program has to be able to identify who prepared the slides. Reni J. Rebecca Barnhart wrote: Recently we started monitoring slide quality in the histology department. We have noticed a trend of folds/wrinkles. I have changed paraffins from Surgipath Blue Ribbon to Fisher's Paraplast X-tra and it does not seem to have made a difference. I have tired using Halt which didn't make a difference either. Most of our specimens are GI biopsies, curettings, skins and breast biopsies. Any other suggestions? Thanks for the help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. ------------------------------ Message: 2 Date: Fri, 30 Nov 2007 11:18:21 -0500 From: "Douglas D Deltour" Subject: RE: SPAM-LOW: [Histonet] Slide quality To: "'Rebecca Barnhart'" , Message-ID: Content-Type: text/plain; charset="US-ASCII" Rebecca, I would defiantly check to see if the wrinkles are generated by one particular person. My techs initial the slides that they cut. This is the easiest way to see if things of this nature are a trend. Douglas D. Deltour HT(ASCP) Histology Manager Professional Pathology Services, PC One Science Court Suite 200 Columbia, SC 29203 Office (803)252-1913 Fax (803)254-3262 Doug@ppspath.com ***************************************************** PROFESSIONAL PATHOLOGY SERVICES, PC NOTICE OF CONFIDENTIALITY This message is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication is strictly prohibited by law. If you have received this communication in error, please notify me immediately. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rebecca Barnhart Sent: Friday, November 30, 2007 10:53 AM To: histonet@lists.utsouthwestern.edu Subject: SPAM-LOW: [Histonet] Slide quality Recently we started monitoring slide quality in the histology department. We have noticed a trend of folds/wrinkles. I have changed paraffins from Surgipath Blue Ribbon to Fisher's Paraplast X-tra and it does not seem to have made a difference. I have tired using Halt which didn't make a difference either. Most of our specimens are GI biopsies, curettings, skins and breast biopsies. Any other suggestions? Thanks for the help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Fri, 30 Nov 2007 08:31:25 -0800 (PST) From: Stacey Burton Subject: [Histonet] remove from list To: Histonet@lists.utsouthwestern.edu Message-ID: <630300.59097.qm@web30007.mail.mud.yahoo.com> Content-Type: text/plain; charset=us-ascii Please remove me from the mailing list. Thank you, Stacey Burton ____________________________________________________________________________ ________ Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. http://mobile.yahoo.com/sports;_ylt=At9_qDKvtAbMuh1G1SQtBI7ntAcJ ------------------------------ Message: 4 Date: Fri, 30 Nov 2007 11:31:06 -0500 From: "Disher Lori" Subject: [Histonet] Decal Solutions and Her2 Fish To: Message-ID: <095327C7CDBDF64B9E9728A54799091E33A593@ORLEV03.hca.corpad.net> Content-Type: text/plain; charset="iso-8859-1" Hello, Can anyone recomend a decal solution to use that will not destroy the cells for HER 2 Fish? If so, do you have a policy? Lori Disher Fawcett Memorial Hosp 21298 Olean Blvd Port Charlotte, FL 33952 941-627-6128 ------------------------------ Message: 5 Date: Fri, 30 Nov 2007 09:25:31 -0800 (PST) From: Irene Cochran Subject: [Histonet] sudan black stain for fecal fat To: histonet@lists.utsouthwestern.edu Message-ID: <746750.38134.qm@web45608.mail.sp1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Our pathologist is looking for a procedure on staining fecal fat with sudan black any suggestions? thanks --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. ------------------------------ Message: 6 Date: Fri, 30 Nov 2007 12:33:53 -0500 From: "Piche-Grocki, Jessica" Subject: [Histonet] CDX2 and HCA (hepatocellular carcinoma) To: Message-ID: <0A57D6AEAE4CBA4A984D27257160A72D01069D0B@win03exchange01.wtbyhosp.org> Content-Type: text/plain; charset="iso-8859-1" Hello, I am having a hard time finding information for our pathologists on CDX2 and HCA antibodies. If anyone has any information they can share that would be great. I am especially having a hard time with the HCA(hepatocellular carcinoma antigen). Thanks and have a great weekend!! Jessica Piche-Grocki, HT(ASCP) CONFIDENTIALITY NOTICE: This email and any attachments contain confidential information that is legally privileged. This information is intended only for the use of the individual or entity named above. The authorized recipient of this information is prohibited from disclosing this information to any other party unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution or action taken in reliance on the contents of these documents is strictly prohibited. If you have received this information in error, please notify the sender immediately and delete these documents. Copyright (c) Waterbury Hospital ------------------------------ Message: 7 Date: Fri, 30 Nov 2007 09:39:42 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Decal Solutions and Her2 Fish To: Disher Lori , histonet@lists.utsouthwestern.edu Message-ID: <916852.26259.qm@web61213.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 We used RDO and EDTA for more delicate specimens (and BM) and after that we regularly did IHC on those sections. I don't think that Her2 neu FISH would behave any differently, although I never did that (FISH always on breast tumors not requiring decal). Reni J. Disher Lori wrote: Hello, Can anyone recomend a decal solution to use that will not destroy the cells for HER 2 Fish? If so, do you have a policy? Lori Disher Fawcett Memorial Hosp 21298 Olean Blvd Port Charlotte, FL 33952 941-627-6128 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better sports nut! Let your teams follow you with Yahoo Mobile. Try it now. ------------------------------ Message: 8 Date: Fri, 30 Nov 2007 09:41:57 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] sudan black stain for fecal fat To: Irene Cochran , histonet@lists.utsouthwestern.edu Message-ID: <57495.58444.qm@web61216.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 And don't you use Oil Red O? You have not processed the feces and could use the ORO Reni J. Irene Cochran wrote: Our pathologist is looking for a procedure on staining fecal fat with sudan black any suggestions? thanks --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get easy, one-click access to your favorites. Make Yahoo! your homepage. ------------------------------ Message: 9 Date: Fri, 30 Nov 2007 12:49:26 -0500 From: Lorraine Cornett Subject: [Histonet] Amyloid To: Histonet Listserve Message-ID: Content-Type: text/plain; charset="iso-8859-1" Thanks to everyone on the histonet, I have a couple of control blocks on the way! Have a great weekend!Lorraine Cornett, HT (ASCP)Highlands PathologyBlue Ridge Division, Kingsport, TN423 224-5793 fax 423 224-5349 _________________________________________________________________ Your smile counts. The more smiles you share, the more we donate. Join in. www.windowslive.com/smile?ocid=TXT_TAGLM_Wave2_oprsmilewlhmtagline ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 48, Issue 42 **************************************** From Janet.Bonner <@t> FLHOSP.ORG Fri Nov 30 12:12:58 2007 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Nov 30 12:14:23 2007 Subject: [Histonet] Re: question about coating blocks with paraffinforstorage References: Message-ID: <5F31F38C96781A4FBE3196EBC22D47802E11D0@fhosxchmb006.ADVENTISTCORP.NET> ...and we have tissue-eating mice. -Florida ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rittman, Barry R Sent: Thu 11/29/2007 1:44 PM To: Rene J Buesa; Robert Richmond; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Re: question about coating blocks with paraffinforstorage It is not only insects but also the possibility of various fungi invading the tissue. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Thursday, November 29, 2007 12:37 PM To: Robert Richmond; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Re: question about coating blocks with paraffin forstorage But I did in Cuba, and there were also "FFPE" tissue eating cockroaches (although perhaps not now, they probably have died of lack of food!). Ren? J. Robert Richmond wrote: The late great R.D. Lillie, who spent his later years at LSU, pointed out that in the warm wet climate of Louisiana if you didn't seal your paraffin blocks the cockroaches would eat the tissue out of them. I've never seen this, but I've never practiced in Louisiana either. Bob Richmond Samurai Pathologist Knoxville TN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Be a better pen pal. Text or chat with friends inside Yahoo! Mail. See how. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ======================================================= The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or 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 the sender immediately by replying to this message and deleting the material from any computer. ======================================================= From MICHELLE.MCNEESE <@t> childrens.com Fri Nov 30 12:24:31 2007 From: MICHELLE.MCNEESE <@t> childrens.com (MICHELLE MCNEESE) Date: Fri Nov 30 12:24:58 2007 Subject: [Histonet] Tissue-Tek Quick-Ray Message-ID: <475000FF020000290000E33F@CNET3.CHILDRENS.COM> Our lab is looking into doing Tissue Microarrays. Because some of them are cost prohibitive, we have been investigating the Tissue-Tek Quick Ray. Does anybody have experience with this particular model? If so, could you please let me know what you like and do not like about it. Thank you! Michelle L McNeese, HT (ASCP) Lead Tech, Histology Department of Pathology Childrens Medical Center (214) 456-6146 Email: michelle.mcneese@childrens.com From Heather.D.Renko <@t> osfhealthcare.org Fri Nov 30 12:39:16 2007 From: Heather.D.Renko <@t> osfhealthcare.org (Renko, Heather D.) Date: Fri Nov 30 12:40:34 2007 Subject: [Histonet] RE: Retic jargon Message-ID: <40026EDDE64CDA47AB382C52619ACD3C0A09DC4A@pmc-rfd-mx01.intranet.osfnet.org> We say "Reticulum" ??? The term reticulin was coined in 1892 by M. Siegfried.[2] Today the term reticulin or reticular fiber is restricted to fibers composed of type III collagen . However, during the pre-molecular era, there was confusion in the use of the term 'reticulin', which was used to describe two structures: * the argyrophilic (silver staining) fibrous structures present in basement membranes * histologically similar fibers present in developing connective tissue[3] . The history of the reticulin silver stain is reviewed by Puchtler et al. (1978).[4] The abstract of this paper says: Maresch (1905) introduced Bielschowsky's silver impregnation technic for neurofibrils as a stain for reticulum fibers, but emphasized the nonspecificity of such procedures. This lack of specificity has been confirmed repeatedly. Yet, since the 1920's the definition of "reticulin" and studies of its distribution were based solely on silver impregnation technics. The chemical mechanism and specificity of this group of stains is obscure. Application of Gomori's and Wilder's methods to human tissues showed variations of staining patterns with the fixatives and technics employed. Besides reticulum fibers, various other tissue structures, e.g. I bands of striated muscle, fibers in nervous tissues, and model substances, e.g. polysaccharides, egg white, gliadin, were also stained. Deposition of silver compounds on reticulum fibers was limited to an easily removable substance; the remaining collagen component did not bind silver. These histochemical studies indicate that silver impregnation technics for reticulum fibers have no chemical significance and cannot be considered as histochemical technics for "reticulin" or type III collagen. Who'd a thunk it? TGIF Heather D. Renko, Histology Coordinator OSF Saint Anthony Medical Center 5666 East State Street Rockford, Illinois 61108 Direct: 815-395-5410 Heather.D.Renko@osfhealthcare.org ============================================================================== The information in this message is confidential and may be legally privileged. Access to this message by anyone other than the addressee is not authorized. If you are not the intended recipient, or an agent of the intended recipient, any disclosure, copying, or distribution of the message or any action or omission taken by you in reliance on it, is prohibited and may be unlawful. If you have received this message in error, please contact the sender immediately and permanently delete the original e-mail, attachment(s), and any copies. ============================================================================== From oshel1pe <@t> cmich.edu Fri Nov 30 12:41:15 2007 From: oshel1pe <@t> cmich.edu (Philip Oshel) Date: Fri Nov 30 12:41:31 2007 Subject: [Histonet] Re: question about coating blocks with paraffinforstorage In-Reply-To: <5F31F38C96781A4FBE3196EBC22D47802E11D0@fhosxchmb006.ADVENTISTCORP.NET> References: <5F31F38C96781A4FBE3196EBC22D47802E11D0@fhosxchmb006.ADVENTISTCORP.NET> Message-ID: A little breeding and you could market the first Biotome. Phil >...and we have tissue-eating mice. -Florida > >________________________________ > >From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rittman, Barry R >Sent: Thu 11/29/2007 1:44 PM >To: Rene J Buesa; Robert Richmond; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Re: question about >coating blocks with paraffinforstorage > > > >It is not only insects but also the possibility >of various fungi invading the tissue. >Barry > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] >On Behalf Of Rene J Buesa >Sent: Thursday, November 29, 2007 12:37 PM >To: Robert Richmond; histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Re: question about >coating blocks with paraffin forstorage > >But I did in Cuba, and there were also "FFPE" >tissue eating cockroaches (although perhaps not >now, they probably have died of lack of food!). > Ren? J. > >Robert Richmond wrote: > The late great R.D. Lillie, who spent his later years at LSU, pointed >out that in the warm wet climate of Louisiana if you didn't seal your >paraffin blocks the cockroaches would eat the tissue out of them. I've >never seen this, but I've never practiced in Louisiana either. > >Bob Richmond >Samurai Pathologist >Knoxville TN -- Philip Oshel Microscopy Facility Supervisor Biology Department 024C Brooks Hall Central Michigan University Mt. Pleasant, MI 48859 (989) 774-3576 From Tiffany.Price <@t> thomaswv.org Fri Nov 30 12:44:52 2007 From: Tiffany.Price <@t> thomaswv.org (Price, Tiffany) Date: Fri Nov 30 12:49:11 2007 Subject: [Histonet] Bridge for Meditech Client Server/Magic Pathology Modules Message-ID: <7B5F5D9A00739F43A1819403EC7CF49103C3248C@thm-mail.thomaswv.org> Does anyone know where I might get information that could help with linking Meditech Client Server with Meditech Magic Pathology modules? I need to be able to get patient demographic information/ previous cases from one facility to the other without manually copying requisitions and face sheets. Thanks Tiffany Confidentiality Notice: This email message including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privilege 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 message. From marybeth.kaulahao <@t> thibodaux.com Fri Nov 30 13:07:04 2007 From: marybeth.kaulahao <@t> thibodaux.com (Marybeth Kaulahao) Date: Fri Nov 30 13:07:08 2007 Subject: [Histonet] pLEASE REMOVE ME Message-ID: <002001c83384$31e28e90$7496010a@trmc.thibodaux.com> Please remove me from the mailing list. Second request. Mary Beth Kaulahao From Sharon.Cook <@t> osumc.edu Fri Nov 30 13:15:11 2007 From: Sharon.Cook <@t> osumc.edu (Cook, Sharon ) Date: Fri Nov 30 13:15:16 2007 Subject: [Histonet] OSUMC Clinical Histology Manager position available. Message-ID: <7A541592F9A53A4E86E7A3D5C4C7F68C01D794ED@msxc01.osumc.edu> The Department of Pathology at The Ohio State University Medical Center is seeking a lab professional for a full time Histology Manager position. This individual will be responsible for the management of Histology and will direct and monitor all personnel, technical and quality assurance activities, and maintain lab adherence to all regulatory (CLIA, CAP, OSHA) standards, as well as monitor and prepare budget, select personnel and develop and maintain monthly productivity benchmarking. The laboratory is a state-of-the-art facility offering a full range of clinical laboratory services. We offer a competitive salary in addition to a comprehensive employee benefit package featuring health insurance, paid leave program, life insurance, short term and long term disability. The Ohio State University Medical Center System is composed of a 923-bed tertiary care hospital, a 160-bed NCI designated comprehensive cancer center, and a 150-bed community hospital with a pathology residency training program, dermatopathology fellowship program (beginning July 2008), Pathologist Assistant Masters program, and clinical histology training site for several online Histology programs. Last year, approximately 65,000 surgical cases were processed with approximately 11,000 HC stains. The laboratory staff consists of 12 HTs, 4 Lab Assistants, 2 Students, and Quality / Technical Training Manager. The Ohio State University Medical Center is located in Columbus, OH home of the Big Ten Ohio State University and located within 2 ? hours of Cincinnati, Cleveland, Pittsburgh and Indianapolis. We require ASCP certification for HT or HTL with a minimum of five years experience in Histology plus strong leadership skills. If you're interested in joining our team, please contact me at the addresss below. Thanks, Sharon Sharon S. Cook, MT(ASCP) SBB Operations Director, Anatomic Pathology The Ohio State University E407 Doan Hall 410 West 10th Avenue Columbus, OH 43210 sharon.cook@osumc.edu Phone: 614 / 293-8418 Fax: 614 / 293-2779 Pager: 614 / 346-0746 From vazquezr <@t> ohsu.edu Fri Nov 30 13:19:39 2007 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Nov 30 13:20:02 2007 Subject: [Histonet] Slide quality Message-ID: Try soaking the faced block in a ammonia water sol for 1 min and in between each level. Robyn OHSU From hfedor <@t> jhmi.edu Fri Nov 30 13:23:59 2007 From: hfedor <@t> jhmi.edu (Helen Fedor) Date: Fri Nov 30 13:24:13 2007 Subject: [Histonet] Tissue-Tek Quick-Ray In-Reply-To: <475000FF020000290000E33F@CNET3.CHILDRENS.COM> References: <475000FF020000290000E33F@CNET3.CHILDRENS.COM> Message-ID: <47501CFF.61A1.0088.3@jhmi.edu> Dear Michelle, I have used this device and it works, it is fairly inexpensive. The TMA blocks section very well. If you are not going to make a lot of arrays then this is definitely a good product for you. It does not come with the ability of doing 0.6mm size punch. So you are limited to the larger size punches. Advantages: Inexpensive compact (you can put it in a drawer when not it use) Ease of sectioning (the sections come off the block like butter) Disadvantages: No high density arrays (0.6mm punch didn't exist when I purchased it) easily fatigued while arraying since you have to hold the device in you hand need to purchase the companies recipient blocks. Helen Helen L. Fedor B.S. Johns Hopkins University Pathology Department 600 N Wolfe St Marburg Room 406 Baltimore MD 21287 email: hfedor@jhmi.edu Phone: 410 614-1660 Pager: 410 283-3419 WARNING: E-mail sent over the Internet is not secure. Information sent by e-mail may not remain confidential. DISCLAIMER: This e-mail is intended only for the individual to whom it is addressed. It may be used only in accordance with applicable laws. If you received this e-mail by mistake, notify the sender and destroy the e-mail. >>> MICHELLE MCNEESE 11/30/2007 1:24 PM >>> Our lab is looking into doing Tissue Microarrays. Because some of them are cost prohibitive, we have been investigating the Tissue-Tek Quick Ray. Does anybody have experience with this particular model? If so, could you please let me know what you like and do not like about it. Thank you! Michelle L McNeese, HT (ASCP) Lead Tech, Histology Department of Pathology Childrens Medical Center (214) 456-6146 Email: michelle.mcneese@childrens.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From amio <@t> stanford.edu Fri Nov 30 13:50:17 2007 From: amio <@t> stanford.edu (Ami Okada) Date: Fri Nov 30 13:50:24 2007 Subject: [Histonet] Leitz 1400 sliding microtome Message-ID: <20071130115017.xzjpifz9noscgs0g@webmail.stanford.edu> Hi THere is a chance that I might be able to get a used Leitz 1400 sliding microtome in apparent good condition for a reasonable price ($1000), with a cooling stage. I was wondering if anyone has had any experience with it, and know whether this is a good price or not. I am also curios as to what type of blade it takes. Thank you so much in advance for your advice -Ami From b-frederick <@t> northwestern.edu Fri Nov 30 14:09:02 2007 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Fri Nov 30 14:09:20 2007 Subject: [Histonet] Slide quality In-Reply-To: Message-ID: <001701c8338c$dd1be510$d00f7ca5@lurie.northwestern.edu> What's a wrinkle? (they're not allowed!!!!) Cutting angle and compression can give wrinkles too. I "grew up" using a 48 degree waterbath and paraplast+, then extra. I only get wrinkles if I get inferior paraffin from one of the outside institutions we deal with. Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Friday, November 30, 2007 1:20 PM To: histonet@lists.utsouthwestern.edu; RBARNHART@summithealth.org Subject: Re: [Histonet] Slide quality Try soaking the faced block in a ammonia water sol for 1 min and in between each level. Robyn OHSU _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From djemge <@t> aol.com Fri Nov 30 14:46:38 2007 From: djemge <@t> aol.com (djemge@aol.com) Date: Fri Nov 30 14:46:47 2007 Subject: [Histonet] Re: Ultramicrotome and other Equipment ie Cryostats Message-ID: <8CA01A93D495BF3-F00-1EA7@Webmail-mg12.sysops.aol.com> You can find used / refurbished?equipment at the following places: Labx.com, Rankin biomedical, Tech One Biomedical, Pmed, eBay, Pacific Southwest?and several other sources. Some great bargains can be found at Labx.com and eBay if you are patient and careful. Donna Emge, HT (ASCP) Northwestern University Feinberg School of Medicine 303 E. Superior St. Lurie 7-220 Chicago, IL 60611 d-emge@northwestern.edu 312-503-2036 ________________________________________________________________________ More new features than ever. Check out the new AOL Mail ! - http://o.aolcdn.com/cdn.webmail.aol.com/mailtour/aol/en-us/text.htm?ncid=aolcmp00050000000003 From alaskagirl1950 <@t> yahoo.com Fri Nov 30 15:27:00 2007 From: alaskagirl1950 <@t> yahoo.com (Patricia Adams) Date: Fri Nov 30 15:27:05 2007 Subject: [Histonet] IHC charges Message-ID: <558551.2170.qm@web52511.mail.re2.yahoo.com> I need to come up with charges for researchers to be able to add to grant money. They will be buying their own antibodies, but I need to give them a charge for the work and chemicals and wash buffers that will be used on the Dako Autostainer plus. Do any of you doing research have a cost worked up per slide? Any help you can give will be very helpful! Thank you in advance. Patricia Adams ----- Fight back spam! Download the Blue Frog. http://www.bluesecurity.com/register/s?user=YWxhc2thZ2lybDE5NTA%3D ____________________________________________________________________________________ Never miss a thing. Make Yahoo your home page. http://www.yahoo.com/r/hs From rjbuesa <@t> yahoo.com Fri Nov 30 15:44:36 2007 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Nov 30 15:44:39 2007 Subject: [Histonet] IHC charges In-Reply-To: <558551.2170.qm@web52511.mail.re2.yahoo.com> Message-ID: <154402.3550.qm@web61220.mail.yahoo.com> Patricia: Under separate cover I am sending my costs analyses. Ren? J. Patricia Adams wrote: I need to come up with charges for researchers to be able to add to grant money. They will be buying their own antibodies, but I need to give them a charge for the work and chemicals and wash buffers that will be used on the Dako Autostainer plus. Do any of you doing research have a cost worked up per slide? Any help you can give will be very helpful! Thank you in advance. Patricia Adams ----- Fight back spam! Download the Blue Frog. http://www.bluesecurity.com/register/s?user=YWxhc2thZ2lybDE5NTA%3D ____________________________________________________________________________________ Never miss a thing. Make Yahoo your home page. http://www.yahoo.com/r/hs _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Never miss a thing. Make Yahoo your homepage. From histotech40 <@t> yahoo.com Fri Nov 30 17:15:20 2007 From: histotech40 <@t> yahoo.com (Sandy Smith) Date: Fri Nov 30 17:15:23 2007 Subject: [Histonet] Slide quality Message-ID: <648380.1758.qm@web57412.mail.re1.yahoo.com> When should the pathologist document wrinkles/folds? Only when it interferes with the diagnosis? Or any time there is a wrinkle or fold? Also how many histo techs out there get 100% wrinkle free sections? I have had discussions with my pathologists about these issues and would like others input. I believe any time there is a wrinkle or fold it should be documented. ____________________________________________________________________________________ Get easy, one-click access to your favorites. Make Yahoo! your homepage. http://www.yahoo.com/r/hs From jnocito <@t> satx.rr.com Fri Nov 30 18:25:28 2007 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Nov 30 18:25:36 2007 Subject: [Histonet] semantics? References: <5AEC610C1CE02945BD63A395BA763EDE011B6FED@NVCIEXCH02.NVCI.org> <019501c832ea$635acbd0$0302a8c0@yourxhtr8hvc4p> <641CEFFC7E5B6C42AB59539653FD082304E0B578@wcu-ex-emp2.PASSHE.LCL> Message-ID: <007e01c833b0$ad4bdd30$0302a8c0@yourxhtr8hvc4p> here, here or is that hear, hear? ----- Original Message ----- From: "Monson, Frederick " To: "Joe Nocito" ; "Tarango, Mark" ; "Tom McNemar" ; "Horn, Hazel V" ; Sent: Friday, November 30, 2007 12:07 PM Subject: RE: [Histonet] semantics? I agree with Joe (here I am butting in again), but..... I like 'dissection' rather than 'disection' which terms some lexicographers like to treat as synonyms. Since I am write and they R rong, and I am in charge, all my minions have been dissectors rather than disectors. I always hope that my surgeon will be the former rather than the latter. Thus it goes with arbitrary, and even sometimes correct, bosses. Even when they are wrong, they assume the right to be left when they are talking to their techs. I assume that many pathologists have bottles of dye/stain on their shelves designated as either 'blue collagen' or 'pink collagen' stain which phrases they must also use in their presentations. In such cases their sub-ordinates merely have to kno the final color to choose the correct one. Of course a peer may ask at a luncheon, "What's the difference between red and pink collagen? Surely there must be a molecular explanation. Have you ever seen yellow collagen or green? Perhaps there is a paper here." I vote for the guy/gal who invented the stain. Surely s/he got it wrong for all of us to follow. Jeers, Cheers, and Beers, it's Friday! Fred Monson Frederick C. Monson, PhD Technical Director Microanalysis and Imaging Research and Training Center (MIRTC) Large Scientific Instrument Core Geology, West Chester University S. Church St. and W. Rosedale Ave. West Chester, PA, 19320 610-738-0437 fmonson@wcupa.edu New Scheduler: http://lexspiac.wcupa.edu/cgi-bin/ureserve_gold.pl Web Page: http://lexspiac.wcupa.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Thursday, November 29, 2007 7:46 PM To: Tarango, Mark; Tom McNemar; Horn, Hazel V; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] semantics? Scotch works for me Joe ----- Original Message ----- From: "Tarango, Mark" To: "Tom McNemar" ; "Horn, Hazel V" ; Sent: Thursday, November 29, 2007 3:57 PM Subject: RE: [Histonet] semantics? I think it's reticulum network, reticular fibers, or reticulin fibers that we're staining. I really don't think that the label needs to be corrected. Mark Adam Tarango HT(ASCP) Histology & IHC Supervisor Nevada Cancer Institute One Breakthrough Way Las Vegas, NV 89135 Direct Line (702) 822-5112 Treo (702) 759-9229 Fax (702) 939-7663 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tom McNemar Sent: Thursday, November 29, 2007 10:01 AM To: Horn, Hazel V; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] semantics? Reticulin here. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Horn, Hazel V Sent: Thursday, November 29, 2007 11:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] semantics? Is it a reticular stain or a reticulum stain? On our labels for the retic stain we put the word, reticulum. One of my pathologists has asked me to correct this with the word, reticular. So, for 35 years have I been using the wrong word? Yikes. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3912 visit us on the web at: www.archildrens.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "EMF " made the following annotations. ------------------------------------------------------------------------ ------ CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential, proprietary, and/or privileged information protected by law. If you are not the intended recipient, you may not use, copy, or distribute this e-mail message or its attachments. If you believe you have received this e-mail message in error, please contact the sender by reply e-mail and destroy all copies of the original message ======================================================================== ====== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet