From timyoon2000 <@t> hotmail.com Mon Jan 2 23:21:36 2006 From: timyoon2000 <@t> hotmail.com (Tim Yoon) Date: Mon Jan 2 23:21:43 2006 Subject: [Histonet] Laser Microdissection services Message-ID: Hello all, Does anyone know of any consultant group that knows how to use laser microdissection technology onto microarray analysis? My company has an Arcturus and a Leica laser microdissection instrument, but due to turnover, nobody knows how to use it. We are trying to find a person (or a group) that can train my team how to take clinical samples onto different microarray platforms using LCM. Thanks in advance Tim Yoon Pfizer _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From tracey.couse <@t> ibb.gatech.edu Tue Jan 3 08:34:47 2006 From: tracey.couse <@t> ibb.gatech.edu (Tracey Couse) Date: Tue Jan 3 08:36:31 2006 Subject: [Histonet] Histologist Position at Gatech - Atlanta, GA Message-ID: <43BA8B87.90500@ibb.gatech.edu> Happy New Year everyone! In case any interested histologists missed this psosting in December... I will soon be leaving my current position as Laboratory Coordinator/Histologist here at Georgia Institute of Technology. I am assisting my supervisor in locating a replacement. If anyone is interested in a technically challenging as well as socially and academically stimulating position with the Gatech/Emory Center for the Engineering of Living Tissues, please read on. The lucky candidate will have the opportunity to work with a variety of personnel (graduate students, staff, and faculty) as well as sample types. The newly renovated (1 year old), fully equipped core lab currently utilizes frozen, paraffin, and plastic techniques. Below is a link to Gatech?s Human Resources website with the official posting. I have also included the formal job description below. If anyone is interested, please contact myself or supervisor with any questions or visit the link below. Sincerely, Tracey -- Tracey Couse Laboratory Coordinator Georgia Tech/Emory Center for the Engineering of Living Tissues Georgia Institute of Technology Office: 404.385.2611 Lab: 404-385-6735 Fax: 404.894.2291 Steve Woodard Laboratory Manager/Safety Officer Petit Institute for Bioengineering and Bioscience Georgia Institute of Technology Email: steve.woodard@ibb.gatech.edu https://ea.ohr.gatech.edu/FullDescription.asp?jobid=CEW4987&type=3&typeofjob=ext&jobtitle=LABORATORY%20COORDINATOR Laboratory Coordinator Institute for Bioengineering and Biosciences Duties: Supervise the histology laboratory and support faculty and graduate student researchers; instruct, perform experiments (e.g. immunohistochemistry); lab safety and compliance knowledge; microscopy knowledge; ability to exchange information with academic/research faculty and graduate information. Position requires strong interpersonal communications skills for advising, exchanging information, teaching; responsibility for promoting an atmosphere of acceptance and respect among employees and customers. Education: A Bachelor?s Degree, preferably in Chemistry or Biochemistry, or any equivalent combination of education and experience is required. Experience: Three years of job related experience is required; preferred computer experience to include Windows, Office 2000/PC. -- Tracey Couse Laboratory Coordinator Georgia Tech/Emory Center for the Engineering of Living Tissues Georgia Institute of Technology IBB, Room 1123 Office: 404.385.2611 Lab: 404-385-6735 Fax: 404.894.2291 From benoit.delatour <@t> ibaic.u-psud.fr Tue Jan 3 10:09:31 2006 From: benoit.delatour <@t> ibaic.u-psud.fr (=?iso-8859-1?Q?Beno=EEt?= Delatour) Date: Tue Jan 3 09:09:19 2006 Subject: [Histonet] Thalamic lesions in Alzheimer mice Message-ID: <6.0.1.1.2.20060103155559.02972ec0@mailhost.pop.u-psud.fr> Dear all, First of all, happy new year! I was wondering if one of you may have an idea concerning a very curious lesion we recently observed in the thalamus of "Alzheimer" (APP) transgenic mice. Most specifically these lesions are located in the ventral posterior complex (Ventral postero lateral (VPL) and Ventral postero medial (VPM) nuclei). They do not seem to be visible in other brain regions. Results from neuropathological examination are the following: -These lesions are mainly associated with VPL/VPM amyloid deposits (plaques) and have a cristal-like appearance with a Nissl stain. -They are positive for iron (PERLS-DAB) and calcium (Alizarine red) -They do not seem to be associated with vascular abnormalities (fact. VIII IHC) or enhanced inflammatory reaction (lectin IHC). If someone has been confronted with similar observations or has any ideas/suggestions concerning the origin / pathological mechanism associated to / identification of such neuropathological alterations... (I can send microphotos to anyone interested). Thanks in advance! B. Delatour _______________________________ B. Delatour Laboratoire de Neurobiologie de l'Apprentissage, de la M?moire & de la Communication, NAMC, CNRS UMR 8620, B?t 446 Universit? Paris-Sud 91405 Orsay Cedex, FRANCE Email benoit.delatour@ibaic.u-psud.fr Web [1]http://www.namc.u-psud.fr References 1. 3D"http://www.namc.u-psud.fr/" From Jeffery-Smith <@t> ouhsc.edu Tue Jan 3 09:27:45 2006 From: Jeffery-Smith <@t> ouhsc.edu (Smith, Jeffery D. (HSC)) Date: Tue Jan 3 09:27:54 2006 Subject: [Histonet] RE:Happy or Merry Message-ID: Sorry for the late reply, but I have been out of the office for a week and a half. All Holiday greetings should be perfectly acceptable to the Christians. After all "holiday" is derived from the the Biblical Holy Day. God said remember the Sabbath to keep it Holy. As this is translated to mean a day of not working, it follows that the earliest days taken off of work were termed Holy Days and are still referred to as holidays. From sjchtascp <@t> yahoo.com Tue Jan 3 09:37:41 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Tue Jan 3 09:37:50 2006 Subject: [Histonet] HT Position Message-ID: <20060103153741.57283.qmail@web90205.mail.scd.yahoo.com> I'm Looking for an HT position in So.WI or No. Ill. --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less From Sandra.Harrison3 <@t> va.gov Tue Jan 3 09:57:08 2006 From: Sandra.Harrison3 <@t> va.gov (Harrison, Sandra C.) Date: Tue Jan 3 09:57:22 2006 Subject: [Histonet] optimal thickness for cutting of IHC sections Message-ID: <736E8889E98B8F4FBBD29FDFEC8074BA49BF6E@VHAV23MSGA2.v23.med.va.gov> Hi Histonetters, Is 5 microns the norm for IHC sections? What is considered optimal? Thanks, Sandy From rjbuesa <@t> yahoo.com Tue Jan 3 10:12:16 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Jan 3 10:12:24 2006 Subject: [Histonet] optimal thickness for cutting of IHC sections In-Reply-To: <736E8889E98B8F4FBBD29FDFEC8074BA49BF6E@VHAV23MSGA2.v23.med.va.gov> Message-ID: <20060103161216.47964.qmail@web61220.mail.yahoo.com> Sandy: A 5 ?m thichness is the general standard for most laboratories and we used it, BUT (there is always a "but" isn't it) if we were targeting a nuclear epitone we tried a little thicker (6-7 ?m) and for cytoplasmic ones we tried somewhat thinner (ca. 4 ?m). This approach was also used for histochemistry procedures: at least 6-7 ?m for reticulum staining in bone marrows. I hope this will help you! Ren? J. "Harrison, Sandra C." wrote: Hi Histonetters, Is 5 microns the norm for IHC sections? What is considered optimal? Thanks, Sandy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From japoteete <@t> saintfrancis.com Tue Jan 3 11:50:12 2006 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Tue Jan 3 11:51:18 2006 Subject: [Histonet] optimal thickness for cutting of IHC sections Message-ID: We use 4 ?m for everything unless the pathologist wants a thinner section. I haven't cut a thicker section in over 5 years. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com -----Original Message----- From: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: Tuesday, January 03, 2006 10:12 AM To: Harrison, Sandra C.; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] optimal thickness for cutting of IHC sections Sandy: A 5 ?m thichness is the general standard for most laboratories and we used it, BUT (there is always a "but" isn't it) if we were targeting a nuclear epitone we tried a little thicker (6-7 ?m) and for cytoplasmic ones we tried somewhat thinner (ca. 4 ?m). This approach was also used for histochemistry procedures: at least 6-7 ?m for reticulum staining in bone marrows. I hope this will help you! Ren? J. "Harrison, Sandra C." wrote: Hi Histonetters, Is 5 microns the norm for IHC sections? What is considered optimal? Thanks, Sandy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From pruegg <@t> ihctech.net Tue Jan 3 12:04:32 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Tue Jan 3 12:04:12 2006 Subject: [Histonet] optimal thickness for cutting of IHC sections In-Reply-To: Message-ID: <200601031804.k03I41Op004348@chip.viawest.net> I come from a Hematopathology background where thinner is better, but for general purposes especially for IHC I cut 4 micron thick sections. I find that tissues thinner than 3 microns can stain very pale by IHC methods. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Poteete, Jacquie A. Sent: Tuesday, January 03, 2006 10:50 AM To: 'Rene J Buesa'; Harrison, Sandra C.; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] optimal thickness for cutting of IHC sections We use 4 ?m for everything unless the pathologist wants a thinner section. I haven't cut a thicker section in over 5 years. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com -----Original Message----- From: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: Tuesday, January 03, 2006 10:12 AM To: Harrison, Sandra C.; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] optimal thickness for cutting of IHC sections Sandy: A 5 ?m thichness is the general standard for most laboratories and we used it, BUT (there is always a "but" isn't it) if we were targeting a nuclear epitone we tried a little thicker (6-7 ?m) and for cytoplasmic ones we tried somewhat thinner (ca. 4 ?m). This approach was also used for histochemistry procedures: at least 6-7 ?m for reticulum staining in bone marrows. I hope this will help you! Ren? J. "Harrison, Sandra C." wrote: Hi Histonetters, Is 5 microns the norm for IHC sections? What is considered optimal? Thanks, Sandy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From timyoon2000 <@t> hotmail.com Tue Jan 3 12:06:00 2006 From: timyoon2000 <@t> hotmail.com (Tim Yoon) Date: Tue Jan 3 12:06:10 2006 Subject: [Histonet] RE: Laser Microdissection services FOUND In-Reply-To: Message-ID: Thanks to all for your replies. I found a service site that may be of interest. Check this out. I am going to review it with my manager. www.LCMprep.com Tim >From: "Tim Yoon" >To: histonet@lists.utsouthwestern.edu >CC: timyoon2000@hotmail.com >Subject: Laser Microdissection services >Date: Mon, 02 Jan 2006 21:21:36 -0800 > >Hello all, >Does anyone know of any consultant group that knows how to use laser >microdissection technology onto microarray analysis? >My company has an Arcturus and a Leica laser microdissection instrument, >but due to turnover, nobody knows how to use it. We are trying to find a >person (or a group) that can train my team how to take clinical samples >onto different microarray platforms using LCM. > >Thanks in advance > >Tim Yoon >Pfizer > >_________________________________________________________________ >Express yourself instantly with MSN Messenger! Download today - it's FREE! >http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ > _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From PMonfils <@t> Lifespan.org Tue Jan 3 12:09:20 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Tue Jan 3 12:09:33 2006 Subject: [Histonet] optimal thickness for cutting of IHC sections Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171763B@lsexch.lsmaster.lifespan.org> I believe section thickness is less critical for IHC because antibodies are very large molecules that don't penetrate tissue very well, so regardless of the thickness of the section, you are really only staining the exposed surface of the tissue, perhaps to a depth of 2 microns or so. We have verified this by electron microscopy. This is quite different from standard histochemical procedures where a thicker section results in a more intense stain because the small dye molecules penetrate the tissue readily and stain it all the way through. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Harrison, Sandra C. > Sent: Tuesday, January 3, 2006 7:57 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] optimal thickness for cutting of IHC sections > > Hi Histonetters, > > Is 5 microns the norm for IHC sections? What is considered optimal? > > Thanks, > > Sandy > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From DDittus787 <@t> aol.com Tue Jan 3 14:27:14 2006 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Tue Jan 3 14:27:37 2006 Subject: [Histonet] job posting Message-ID: <13c.22c41018.30ec3822@aol.com> I am posting this job opportunity for someone else, so I would appreciate anyone who is interested ,contacting the e-mail at the end of this message. A very lovely woman called today wanting to know if I knew anyone who would be interested in helping to set up and validate a program on the meditech computer system for Blood bank and a Donor center. The job is in California ( good time for east coldsters) to get warm. If you or anyone you know is interested please send your resume to: _roberta.deluca@acs-hcs.com_ (mailto:roberta.deluca@acs-hcs.com) she will then contact you. Thanks Dana From gcallis <@t> montana.edu Tue Jan 3 14:50:03 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Jan 3 14:50:20 2006 Subject: [Histonet] autofluorescence with vibratome/PFA fixed embryonic brain sections Message-ID: <6.0.0.22.1.20060103134406.01b40290@gemini.msu.montana.edu> Ali, Instead of using a FITC conjugated antibody, try a red fluorophore, i.e. rhodamine (RRX from Jackson) or Texas Red. You will have nice contrast with the greenish autofluorescence that tends to be worse with PFA fixation. Do NOT change the fixation, but simply use a different color. You could also use an antibody conjugated to Alexa 555 (Molecular Probes) which is more resistant to photobleaching and extremely bright - it is the equivalent to rhodamine. From:"Ali Moussavi Nik" ---------- Dear All Happy new year I am doing Immunohistochemistry on 4% PFA fixed, vibrotome sectioned embryonic brain. My antibody is FITC conjugated I have a problem with the background green Autoflorescence of the PFA . Can anyone propose me other method of fixation for embryonic brain. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From pruegg <@t> ihctech.net Tue Jan 3 14:51:00 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Tue Jan 3 14:50:39 2006 Subject: [Histonet] Tunnel Message-ID: <200601032050.k03KoSOp029068@chip.viawest.net> I guess the time has come when I have to get back into doing Tunnel for apoptosis. I have avoided this, and most of my customers are satisified with cleaved caspase 3 in place of Tunnel, but more and more are starting to ask for both. Please share your choice of kits for Tunnel these days so I can gear up for this again. I am also interested in knowing how folks deal with the problem of Tunnel not distinquishing between apoptosis and necrosis (at least that is what we found some years ago, hopefully the technology has improved?). The samples I will be dealing with will have both necrosis and apoptosis going on at the same time. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From drgrant <@t> cmh.edu Tue Jan 3 15:05:40 2006 From: drgrant <@t> cmh.edu (Grant, Debra, R) Date: Tue Jan 3 15:06:52 2006 Subject: [Histonet] EM Survey Message-ID: <6F434E27D5DE944B9E898984CADDD1490439B3@exchmail.CMH.Internal> Hi All, It's survey time! Do labs make their own Glutaraldehyde for EM or buy it? If you buy it, which vendor is preferred? Also, do you keep the glutaraldehyde (with the specimen) in the refrigerator until you send it out to the lab for EM work, or can it be left out at room temperature? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 From pathrm35 <@t> adelphia.net Tue Jan 3 15:48:08 2006 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Tue Jan 3 15:48:17 2006 Subject: [Histonet] Biocare Decloaking Chamber for sale Message-ID: <3169651.1136324888446.JavaMail.root@web22> I have a slightly used Biocare Decloaking Chamber I would like to sell. If there is anyone interested please contact me. Thanks, Ron Martin From rjbuesa <@t> yahoo.com Tue Jan 3 16:00:16 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Jan 3 16:00:25 2006 Subject: [Histonet] EM Survey In-Reply-To: <6F434E27D5DE944B9E898984CADDD1490439B3@exchmail.CMH.Internal> Message-ID: <20060103220016.49362.qmail@web61215.mail.yahoo.com> Hi Debra: We used to make our own glutaraldehyde for EM and our own Michel's for immunofluorescence. We did our own EM work (start to photos enlargements) and the specimens were kept in a regular refrigerator (ca. 4?C0 Ren? J. "Grant, Debra, R" wrote: Hi All, It's survey time! Do labs make their own Glutaraldehyde for EM or buy it? If you buy it, which vendor is preferred? Also, do you keep the glutaraldehyde (with the specimen) in the refrigerator until you send it out to the lab for EM work, or can it be left out at room temperature? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From gcallis <@t> montana.edu Tue Jan 3 16:06:54 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Jan 3 16:07:20 2006 Subject: [Histonet] EM Survey In-Reply-To: <6F434E27D5DE944B9E898984CADDD1490439B3@exchmail.CMH.Intern al> References: <6F434E27D5DE944B9E898984CADDD1490439B3@exchmail.CMH.Internal> Message-ID: <6.0.0.22.1.20060103143231.01b36c90@gemini.msu.montana.edu> Ted Pella, concentrated comes in ampules, made up our own for a recipe that fit the project, or any given concentration of glut. . At 02:05 PM 1/3/2006, you wrote: >Hi All, > >It's survey time! Do labs make their own Glutaraldehyde for EM or buy >it? If you buy it, which vendor is preferred? Also, do you keep the >glutaraldehyde (with the specimen) in the refrigerator until you send it >out to the lab for EM work, or can it be left out at room temperature? > >Kind Regards, > > >Debby R. Grant HT(ASCP) >Histology Coordinator >The Children's Mercy Hospital & Clinics >2401 Gillham Rd. >Kansas City, Missouri 64108 >lab (816)234-3827 >fax (816) 802-1492 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From pruegg <@t> ihctech.net Tue Jan 3 16:10:52 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Tue Jan 3 16:11:16 2006 Subject: [Histonet] Tunnel responses from IHCRG Message-ID: <200601032210.k03MAqOp025529@chip.viawest.net> I recently used the kit from Chemicon cat# S7101 ( Looks just like the old Intergen one to me) and ran caspase 3 along side with near perfect correlation.I was thrilled because I recalled Tunnel as a troublesome assay...I bet its been 5 years since I ran it.I just followed the insert and had no problems. I'd wish you good luck , but you won't need it! Sue Wells Hi Patsy - I use chemicon [a division of serologicals] #S7101. It's a little pricey, but works well. I am willing to share my optimized protocol with you if you choose this one. I do it several times a month for various investigators. Most have no trouble distinguishing necrosis from apoptosis... They look for nuclear "blebbing" and fragmented nuclei, etc. Most requests come from people who have been doing this a long time. [I'm glad I don't have to do this part] Please let me know if you have any questions. Best Regards, Mary Vaughan HT (ASCP) Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From ndruckenbrod <@t> wisc.edu Tue Jan 3 16:41:10 2006 From: ndruckenbrod <@t> wisc.edu (NOAH R DRUCKENBROD) Date: Tue Jan 3 16:41:20 2006 Subject: [Histonet] TUNEL Message-ID: Hi Everyone, I am going to begin some TUNEL experiments for the first time and have some basic questions. First, how difficult is it to stain un-sectioned embryonic tissue? The target cells are about 4-5 cell layers in from the surface. Second, the target cells are transgenically labeled with YFP--will the YFP signal,or its epitope, be lost by the tissue prep for TUNEL? Lastly, could I get some perspective on good/bad TUNEL kits? I have found some of this topic in the archive, but this advice may have a shelf life. Thanks in advance. Cordially, Noah Druckenbrod University of Wisconsin-Madison From AnthonyH <@t> chw.edu.au Tue Jan 3 17:06:16 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Jan 3 17:06:35 2006 Subject: [Histonet] EM Survey Message-ID: We prepare our own Glutaraldehyde for EM and store specimens in it in the fridge before processing. 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 Grant, Debra, R Sent: Wednesday, 4 January 2006 8:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] EM Survey Hi All, It's survey time! Do labs make their own Glutaraldehyde for EM or buy it? If you buy it, which vendor is preferred? Also, do you keep the glutaraldehyde (with the specimen) in the refrigerator until you send it out to the lab for EM work, or can it be left out at room temperature? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ 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 lsunhwa <@t> gmail.com Tue Jan 3 17:53:08 2006 From: lsunhwa <@t> gmail.com (Sunhwa Lee) Date: Tue Jan 3 17:53:15 2006 Subject: [Histonet] Plastic sectioning Message-ID: <177172430601031553h77fb0156hfae3b6942eb39dd0@mail.gmail.com> Hi all I am trying to cut 20-30um plastic sections of Zebrafish brain. The fish was perfused with AFA (alcohol-formalin-acetic acid) for more than a month and treated with Technovit 7100 for plastic sections. Then the sections were stained with Thionin. (Thionin 15m ? running tap water 5m ? rinse in dH2O ? Dry ? DePex) But it got shattering and lots of knife marks on it. Any suggestion for embedding and staining will be very appreciated. Best, Sunny From gcallis <@t> montana.edu Tue Jan 3 18:13:21 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Jan 3 18:13:37 2006 Subject: [Histonet] TUNEL In-Reply-To: References: Message-ID: <6.0.0.22.1.20060103170330.01b4bfd8@gemini.msu.montana.edu> YFP is yellow fluorescent protein, is a chimera of Green fluorescent protein, the barrel shaped protein so popular these days. I am not sure it can be referred to as an epitope, unless my definition of epitope is not complete or you are you are planning to detect the YFP with an antiYFP antibody. Maybe I am missing something on the definition of epitope. Being a chimera of GFP, it may be highly affected by fixation and other chemicals, temperatures, enzymes, even more so than eGFP itself, when used with TUNEL tissue preparations. You can read up on working with GFP and YFP at the Clontech website - where you can access their manual on Living Colours. I think there are now books written on the subject of using GFP and its chimeras. I am not sure one can combine the use of YFP with TUNEL and get the best of both worlds, but a literature search in PUBMED may be helpful. At 03:41 PM 1/3/2006, you wrote: >I am going to begin some TUNEL experiments for the first time and have >some basic questions. First, how difficult is it to stain un-sectioned >embryonic tissue? The target cells are about 4-5 cell layers in from >the surface. Second, the target cells are transgenically labeled with >YFP--will the YFP signal,or its epitope, be lost by the tissue prep for >TUNEL? Lastly, could I get some perspective on good/bad TUNEL kits? I >have found some of this topic in the archive, but this advice may have a >shelf life. Thanks in advance. > >Cordially, >Noah Druckenbrod >University of Wisconsin-Madison Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From cbass <@t> bidmc.harvard.edu Tue Jan 3 21:24:34 2006 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Tue Jan 3 21:24:32 2006 Subject: [Histonet] brain fixation and native EGFP fluorescence Message-ID: Hello Everyone, I have been using a viral vector to express EGFP in the mouse liver and examining native fluorescence. I generally fix the tissue by cutting the liver into blocks, immersing in NBF and sectioning on a cryostat. I have gotten some really good results just looking at native fluorescence. I am hoping to extend this to the brain. I have now injected my viral vector into the striatum of a mouse and am set to collect the tissue. Since I am working with the brain, my natural tendency is to perfuse, as I have always heard this is the best method of fixing brain tissue. Does anyone have advice in terms of picking a good fixative for perfusion? I have been reading about a lot of different fixatives for the brain, one which includes sucrose. I am familiar with sucrose cryopreservation, but I haven't considered adding it to the fix. Any suggestions would be appreciated. If you have a suggestion please include the formula. Someone recently suggested a paraformaldehyde zinc fix to me. What are the advantages of this paraformaldehyde alone and does anyone have a formulation for it? Thanks, Caroline Bass From jkiernan <@t> uwo.ca Tue Jan 3 23:44:16 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Tue Jan 3 23:43:55 2006 Subject: [Histonet] brain fixation and native EGFP fluorescence References: Message-ID: <43BB60B0.9A1CCFE7@uwo.ca> A colleague who knows more than I about these matters tells me that the native fluorescence of GFP depends critically on the highly organized barrel-shaped configuration of the molecule - something that may be preserved or destroyed by fixation, but won't survive dehydration in alcohols etc. Remember that aldehyde-fixed and even unfixed tissues can have green autofluorescence when exited by blue light. Heavy metal salts generally tend to suppress fluorescence, even though some histochemical fluorescence methods can be enhanced by zinc or aluminium salts. It might be best to avoid zinc formaldehyde mixtures for this reason, and also because zinc ions coagulate proteins and can be expected to change the shape of the green fluorescent protein molecule, perhaps making it no longer fluorescent. John Kiernan Anatomy, UWO London, Canada -------------------------- Caroline Bass wrote: > > Hello Everyone, > > I have been using a viral vector to express EGFP in the mouse liver > and examining native fluorescence. I generally fix the tissue by > cutting the liver into blocks, immersing in NBF and sectioning on a > cryostat. I have gotten some really good results just looking at > native fluorescence. I am hoping to extend this to the brain. I > have now injected my viral vector into the striatum of a mouse and am > set to collect the tissue. Since I am working with the brain, my > natural tendency is to perfuse, as I have always heard this is the > best method of fixing brain tissue. > > Does anyone have advice in terms of picking a good fixative for > perfusion? I have been reading about a lot of different fixatives > for the brain, one which includes sucrose. I am familiar with > sucrose cryopreservation, but I haven't considered adding it to the fix. > > Any suggestions would be appreciated. If you have a suggestion > please include the formula. Someone recently suggested a > paraformaldehyde zinc fix to me. What are the advantages of this > paraformaldehyde alone and does anyone have a formulation for it? > > Thanks, > > Caroline Bass > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From patrick.howorth <@t> bristol.ac.uk Wed Jan 4 04:25:24 2006 From: patrick.howorth <@t> bristol.ac.uk (PW Howorth, Physiology) Date: Wed Jan 4 04:25:37 2006 Subject: [Histonet] alexa fluors Message-ID: <9E52F7E00129B8D685A25392@pys-aep6.pys.bris.ac.uk> Hi all, I need to do some triple localisation studies. Previously, I have used FITC, Cy3 and AMCA. I have never really liked AMCA that much, fades quickly and the colour is pretty dire. I'm wondering about using an Alexa fluor which excites either in the blue or near red spectrum. Can you recommend one or the other and a reliable supplier ?! Many thanks Patrick ---------------------- Patrick Howorth, Dept of Physiology, University of Bristol, Bristol, BS8 1TD. +44 (0)117 33 17112 patrick.howorth@bristol.ac.uk From BMolinari <@t> heart.thi.tmc.edu Wed Jan 4 05:46:42 2006 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Wed Jan 4 05:51:45 2006 Subject: [Histonet] EM Survey Message-ID: Hi Debby, We make our own glut and store it in the fridge till it is ready to process. We have an in-house EM lab, but ask that samples sent to the lab be shipped in a Styrofoam container w/ a "frigidbrick". Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Grant, Debra, R Sent: Tuesday, January 03, 2006 3:06 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] EM Survey Hi All, It's survey time! Do labs make their own Glutaraldehyde for EM or buy it? If you buy it, which vendor is preferred? Also, do you keep the glutaraldehyde (with the specimen) in the refrigerator until you send it out to the lab for EM work, or can it be left out at room temperature? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Wed Jan 4 08:56:35 2006 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Wed Jan 4 08:57:16 2006 Subject: [Histonet] brain fixation and native EGFP fluorescence Message-ID: <5784D843593D874C93E9BADCB87342AB916A78@tpiserver03.Coretech-holdings.com> See the following link for a through discussion of perfusion, and how to avoid soft tissue shrinkage. http://www.myneurolab.com/global/Manuals/About%20Perfusion.pdf Sucrose should be used as the prewash, not in the fixative, at least until the tissue is mostly fixed. Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Caroline Bass Sent: Tuesday, January 03, 2006 9:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] brain fixation and native EGFP fluorescence Hello Everyone, I have been using a viral vector to express EGFP in the mouse liver and examining native fluorescence. I generally fix the tissue by cutting the liver into blocks, immersing in NBF and sectioning on a cryostat. I have gotten some really good results just looking at native fluorescence. I am hoping to extend this to the brain. I have now injected my viral vector into the striatum of a mouse and am set to collect the tissue. Since I am working with the brain, my natural tendency is to perfuse, as I have always heard this is the best method of fixing brain tissue. Does anyone have advice in terms of picking a good fixative for perfusion? I have been reading about a lot of different fixatives for the brain, one which includes sucrose. I am familiar with sucrose cryopreservation, but I haven't considered adding it to the fix. Any suggestions would be appreciated. If you have a suggestion please include the formula. Someone recently suggested a paraformaldehyde zinc fix to me. What are the advantages of this paraformaldehyde alone and does anyone have a formulation for it? Thanks, Caroline Bass _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Jan 4 09:25:39 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Jan 4 09:26:04 2006 Subject: [Histonet] alexa fluors In-Reply-To: <9E52F7E00129B8D685A25392@pys-aep6.pys.bris.ac.uk> References: <9E52F7E00129B8D685A25392@pys-aep6.pys.bris.ac.uk> Message-ID: <6.0.0.22.1.20060104082150.01b607a0@gemini.msu.montana.edu> There is only one supplier of Alexa fluorophores - Molecular Probes part of Invitrogen, and you can go to their website to find an Alexa which will excite at the wavelength you want to use. Contact their technical services for recommendations on this, they are very helpful. Molecular Probes Handbook is on line, explaining anything you want to know about Alexa fluorophores including their array of Alexa related products. You will not be disappointed. At 03:25 AM 1/4/2006, you wrote: >Hi all, > >I need to do some triple localisation studies. Previously, I have used >FITC, Cy3 and AMCA. I have never really liked AMCA that much, fades >quickly and the colour is pretty dire. I'm wondering about using an Alexa >fluor which excites either in the blue or near red spectrum. Can you >recommend one or the other and a reliable supplier ?! > >Many thanks > >Patrick >---------------------- >Patrick Howorth, >Dept of Physiology, >University of Bristol, >Bristol, >BS8 1TD. >+44 (0)117 33 17112 >patrick.howorth@bristol.ac.uk > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From rschoon <@t> email.unc.edu Wed Jan 4 09:36:53 2006 From: rschoon <@t> email.unc.edu (rschoon) Date: Wed Jan 4 09:40:19 2006 Subject: [Histonet] alexa fluors In-Reply-To: <6.0.0.22.1.20060104082150.01b607a0@gemini.msu.montana.edu> References: <9E52F7E00129B8D685A25392@pys-aep6.pys.bris.ac.uk> <6.0.0.22.1.20060104082150.01b607a0@gemini.msu.montana.edu> Message-ID: <43BBEB95.9000701@email.unc.edu> Gail, Serotec also sells the Alexa fluorophores. Bob Gayle Callis wrote: > There is only one supplier of Alexa fluorophores - Molecular Probes > part of Invitrogen, and you can go to their website to find an Alexa > which will excite at the wavelength you want to use. Contact their > technical services for recommendations on this, they are very > helpful. Molecular Probes Handbook is on line, explaining anything > you want to know about Alexa fluorophores including their array of > Alexa related products. You will not be disappointed. > > At 03:25 AM 1/4/2006, you wrote: > >> Hi all, >> >> I need to do some triple localisation studies. Previously, I have >> used FITC, Cy3 and AMCA. I have never really liked AMCA that much, >> fades quickly and the colour is pretty dire. I'm wondering about >> using an Alexa fluor which excites either in the blue or near red >> spectrum. Can you recommend one or the other and a reliable supplier ?! >> >> Many thanks >> >> Patrick >> ---------------------- >> Patrick Howorth, >> Dept of Physiology, >> University of Bristol, >> Bristol, >> BS8 1TD. >> +44 (0)117 33 17112 >> patrick.howorth@bristol.ac.uk >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Gayle Callis > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjchtascp <@t> yahoo.com Wed Jan 4 10:21:29 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Wed Jan 4 10:21:38 2006 Subject: [Histonet] mallory-Aaan Trichrome Message-ID: <20060104162129.67340.qmail@web90205.mail.scd.yahoo.com> Does anyone know about this pancreas islet cell stain and whether it stains A,B and D cells? Has anyone uses Maximows modification zenkers fixitive for these cells also.? Thanks, Steve --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From gcallis <@t> montana.edu Wed Jan 4 10:34:03 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Jan 4 10:34:28 2006 Subject: [Histonet] alexa fluors In-Reply-To: <43BBEB95.9000701@email.unc.edu> References: <9E52F7E00129B8D685A25392@pys-aep6.pys.bris.ac.uk> <6.0.0.22.1.20060104082150.01b607a0@gemini.msu.montana.edu> <43BBEB95.9000701@email.unc.edu> Message-ID: <6.0.0.22.1.20060104092033.01b431e8@gemini.msu.montana.edu> Hi Bob, Happy New Year to you and yours - will you be in Charleston for the regional meeting this year? So will Diane Sterchi and I, to give workshops! Serotec does sell Alexa conjugates, as does BD Pharmingen and some other companies, but these antibody/Alexa conjugates tend to be monoclonal or polyclonals for more specific staining (i.e. CD markers) along with immunoglobulin isotype- conjugates rather than some of the other Alexa products, i.e. Strepavidin-Alexa Fluors or secondary antibody-Alexa conjugates and their tidy little conjugation kits found at Molecular Probes. The Serotec website was excellent for some interesting shopping, a heck of a lot friendlier than the horrible, lumbering giganticus BD Pharmingen website - our BD rep hates his own website. Gayle From pruegg <@t> ihctech.net Wed Jan 4 10:43:35 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Wed Jan 4 10:43:50 2006 Subject: [Histonet] serca2 Message-ID: <200601041643.k04GhXOp023106@chip.viawest.net> Has anybody tried using Serca2 (sarco/endoplasmic reticulum calcium ATPase type 2) on FFPE human tissue. I have some papers on it but they are using frozen sections or cell preps not ffpe tissues. The paper I have uses FITC IHC for this antibody. Thanks, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From Rcartun <@t> harthosp.org Wed Jan 4 11:19:42 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Wed Jan 4 11:20:18 2006 Subject: [Histonet] BioChem ImmunoSystems Message-ID: Happy New Year everyone! I am trying to locate a company in Montreal, Canada by the name of "BioChem International". It used to be called IAF BioChem. Can anyone help me? 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 rjbuesa <@t> yahoo.com Wed Jan 4 11:37:38 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Jan 4 11:37:46 2006 Subject: [Histonet] mallory-Aaan Trichrome In-Reply-To: <20060104162129.67340.qmail@web90205.mail.scd.yahoo.com> Message-ID: <20060104173738.20458.qmail@web61212.mail.yahoo.com> Hi Steve: Yes, Mallory-Azan stains α cells RED, ? cells YELLOW, and delta cells BLUE; many years ago I used this stain as well as Maximov's modification (1909) of Zenker's. Formula: water → 250 mL + mercuric chloride → 12.5 g + potassium dichromate → 6.25 g + 40% formaldehyde → 25 mL + sodium sulfate → 2.5 g Ren? J. Steven Coakley wrote: Does anyone know about this pancreas islet cell stain and whether it stains A,B and D cells? Has anyone uses Maximows modification zenkers fixitive for these cells also.? Thanks, Steve --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From mward <@t> wfubmc.edu Wed Jan 4 11:49:46 2006 From: mward <@t> wfubmc.edu (Martha Ward) Date: Wed Jan 4 11:49:57 2006 Subject: [Histonet] cellmarque pressure cooker Message-ID: <61135F0455D33347B5AAE209B903A30411A8FFD5@EXCHVS2.medctr.ad.wfubmc.edu> I hope someone can help me. We received an electric Princess Pressure cooker, model DYB 350 when we ordered some Trilogy a while back. I need to find a replacement gasket but have been unable to find the company (Princess) online so I can get this part. Has anyone else had this problem? CellMarque no longer carries these cookers and was unable to help me. I hate to purchase a new pressure cooker when this one doing fine for us, except for the gasket needing to be replaced. Thanks in advance for any help ya'll can give me. Martha Ward Wake Forest University Baptist Medical Center From Malcolm.McCallum <@t> tamut.edu Wed Jan 4 12:11:36 2006 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Wed Jan 4 12:14:48 2006 Subject: [Histonet] cellmarque pressure cooker Message-ID: I haven't seen one of these, but I have a suggestion that might help you. At auto parts stores (like autozone) they sell stuff for replacing gaskets in automobiles. It comes in a tube like glue. Figure that automobiles operate at high heat and pressures, maybe it will work. Worse case scenario, you might find yourself scraping of the gasket, which frankly isn't that big of a deal once it is dry. Maybe it will work? Anyone out there familiar with the product I am refering to you might suggest a yes or no! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Martha Ward Sent: Wed 1/4/2006 11:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cellmarque pressure cooker I hope someone can help me. We received an electric Princess Pressure cooker, model DYB 350 when we ordered some Trilogy a while back. I need to find a replacement gasket but have been unable to find the company (Princess) online so I can get this part. Has anyone else had this problem? CellMarque no longer carries these cookers and was unable to help me. I hate to purchase a new pressure cooker when this one doing fine for us, except for the gasket needing to be replaced. Thanks in advance for any help ya'll can give me. Martha Ward Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Paula.F.Conlon <@t> Lahey.org Wed Jan 4 12:45:54 2006 From: Paula.F.Conlon <@t> Lahey.org (Conlon, Paula F.) Date: Wed Jan 4 12:45:18 2006 Subject: [Histonet] Frozen Section for Muscles Message-ID: Having a problem with ice crystals showing up on a special stains done on frozen sectoin slides. Moisture is coming from a few sources. We are trying to eliminate these sources. Any suggestions as how to get rid of these crystals would be appreciated. See our web page at http://www.lahey.org for a full directory of Lahey sites, staff, services and career opportunities. THIS MESSAGE IS INTENDED FOR THE USE OF THE PERSON TO WHOM IT IS ADDRESSED. IT MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If you are not the intended recipient, your use of this message for any purpose is strictly prohibited. If you have received this communication in error, please delete the message and notify the sender so that we may correct our records. From mauger <@t> email.chop.edu Wed Jan 4 13:15:42 2006 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Wed Jan 4 13:16:46 2006 Subject: [Histonet] cellmarque pressure cooker Message-ID: Martha, Biocare sells a pressure cooker and replacement gaskets for it. Jo >>> "Martha Ward" 01/04/06 12:49 PM >>> I hope someone can help me. We received an electric Princess Pressure cooker, model DYB 350 when we ordered some Trilogy a while back. I need to find a replacement gasket but have been unable to find the company (Princess) online so I can get this part. Has anyone else had this problem? CellMarque no longer carries these cookers and was unable to help me. I hate to purchase a new pressure cooker when this one doing fine for us, except for the gasket needing to be replaced. Thanks in advance for any help ya'll can give me. Martha Ward Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rachael_Emerson <@t> URMC.Rochester.edu Wed Jan 4 13:43:32 2006 From: Rachael_Emerson <@t> URMC.Rochester.edu (Emerson, Rachael) Date: Wed Jan 4 13:43:59 2006 Subject: [Histonet] ICAM-1 Message-ID: Hello. Does anyone have information/pictures off the staining pattern of ICAM-1 in adult mouse liver? Thank you Rachael -- From dellav <@t> musc.edu Wed Jan 4 14:18:01 2006 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Wed Jan 4 14:18:43 2006 Subject: [Histonet] Frozen Section for Muscles Message-ID: Ice crystals are often the result of one's technique in freezing the sample. you can find two great articles by Gayle Callis in HistoLogic, Spring and Autumn Issues, 2004 www.sakuraus.com/histologic Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Conlon, Paula F." 01/04/06 01:45PM >>> Having a problem with ice crystals showing up on a special stains done on frozen sectoin slides. Moisture is coming from a few sources. We are trying to eliminate these sources. Any suggestions as how to get rid of these crystals would be appreciated. See our web page at http://www.lahey.org for a full directory of Lahey sites, staff, services and career opportunities. THIS MESSAGE IS INTENDED FOR THE USE OF THE PERSON TO WHOM IT IS ADDRESSED. IT MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If you are not the intended recipient, your use of this message for any purpose is strictly prohibited. If you have received this communication in error, please delete the message and notify the sender so that we may correct our records. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anh2006 <@t> med.cornell.edu Wed Jan 4 14:44:00 2006 From: anh2006 <@t> med.cornell.edu (Andrea Hooper) Date: Wed Jan 4 14:44:15 2006 Subject: [Histonet] ICAM-1 Message-ID: Dear Rachael, Based on what I understand about this molecule ICAM-1 should be expressed on the sinusoidal endothelial cells and perhaps the large veins and arteries as well. I would expect in a normal healthy mouse that the expression would be negative/weak. In a mouse undergoing metastatic events or perhaps injury/toxicity to the liver you would expect to see an increase in ICAM-1 expression in these endothelial cells. I am sure a quick search on PubMed would give you just the information you are looking for. Good luck! ----------------------- From: "Emerson, Rachael" ? Sent: Wednesday, January 4, 2006 2:43 pm To: histonet@lists.utsouthwestern.edu? Subject: [Histonet] ICAM-1 Hello. Does anyone have information/pictures off the staining pattern of ICAM-1 in adult mouse liver? Thank you Rachael From AFoshey <@t> chw.org Wed Jan 4 15:50:35 2006 From: AFoshey <@t> chw.org (Foshey, Annette) Date: Wed Jan 4 15:50:54 2006 Subject: [Histonet] Number of GI biopsy sections on a slide for pediatric hospitals. Message-ID: <9E6D52F532809247BDA1783680E92C5658687A@CHWEXC.chwi.chswi.org> I am inquiring what the "standard" is for the number of GI biopsy sections that are routinely put on slides for review. We currently cut two slides with two levels on each slide. The total number of sections is between 6-8 sections on each slide. Thanks, Annette Foshey, HT Team Leader in Histology Children's Hospital of Wisconsin ************************** Children's Hospital and Health System recognizes that unencrypted e-mail is insecure and does not guarantee confidentiality. The confidentiality of replies to this message cannot be guaranteed unless the replies are encrypted. From HornHV <@t> archildrens.org Wed Jan 4 16:00:44 2006 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Wed Jan 4 16:00:49 2006 Subject: [Histonet] Number of GI biopsy sections on a slide for pediatric hospitals. Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDF27@EMAIL.archildrens.org> We cut 2 levels with 3 sections on each slide for GI's. We cut 3 levels on skin and gyns. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail 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 Foshey, Annette Sent: Wednesday, January 04, 2006 3:51 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Number of GI biopsy sections on a slide for pediatric hospitals. I am inquiring what the "standard" is for the number of GI biopsy sections that are routinely put on slides for review. We currently cut two slides with two levels on each slide. The total number of sections is between 6-8 sections on each slide. Thanks, Annette Foshey, HT Team Leader in Histology Children's Hospital of Wisconsin ************************** Children's Hospital and Health System recognizes that unencrypted e-mail is insecure 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 ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From deb.vaneyck <@t> phci.org Wed Jan 4 16:42:30 2006 From: deb.vaneyck <@t> phci.org (Van Eyck, Deb) Date: Wed Jan 4 16:42:44 2006 Subject: [Histonet] Happy New Year's and competancy/volume questions Message-ID: Hi all, If anyone has information they would like to share I would appreciate it. We are a hospital lab and we are updating doing competencies. We are interested in Volumes and expectations for embedding, cutting etc, that take into account differeing experience levels, training and quality expectations---does anyone have a good system or have good references or articles that discuss this issue? thanks for your input. Deb This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. From shu-cheng.chen <@t> spcorp.com Wed Jan 4 17:02:56 2006 From: shu-cheng.chen <@t> spcorp.com (Chen, Shu-Cheng) Date: Wed Jan 4 17:03:48 2006 Subject: [Histonet] postdoctoral position to study psoriasis Message-ID: <886D951F4246D94DAF28C82DC102DDD00C09BB@KENMSG20.us.schp.com> Hi, We have a postdoctoral position available immediately in the Inflammation Department at Schering-Plough to study the pathogenesis of psoriasis. Schering-Plough is a Pharmaceutical company located in Kenilworth NJ, which is within 30 min driving distance from New York city. We are looking for PhD candidates with experience in frozen sections and IHC. The ad has been posted in Nature since Nov 30th 2005 and the link is attached here. http://naturejobs.nature.com/texis/jobsearch/details.html?id=438db2314a01c0& q=postdoc&qField=All&qSort=date&qMatch=all&pp=10&view=1&page=15 . The job code for this particular position is 12243BR. The postdoc will have the opportunity to participate in other drug discovery projects and learning cutting-edge technologies in addition to his/her own project in psoriasis. If you know of any qualified candidates who are looking for an opportunity and the exciting experience to work in the Pharmaceutical industry please pass along this information. Thank you very much for your help! Shu-Cheng Chen ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From rjbuesa <@t> yahoo.com Wed Jan 4 17:32:46 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Jan 4 17:32:53 2006 Subject: [Histonet] Happy New Year's and competancy/volume questions In-Reply-To: Message-ID: <20060104233246.9925.qmail@web61221.mail.yahoo.com> Hi Deb: In 3 labs I wrote competencies for the expectation for sectoning was set at 25 blocks/hour and for embedding at 60 blocks/hour. Each histotech was evaluated on that basis and their individual averages were rated (as percentages) against the expectation. Those with averages above expectations were evaluated better (and received salary compensations accordingly). Nobody was considered "proficient" until their individual average reached the expectation consistently. For time utilization (ratio between time with recorded tasks versus logged time) the expectation was 75 % (i.e. 6 h with logged tasks agains 8 h/day per shift). In the Dec./04 issue of the Journal of Histotechnology there are averages for every imaginable task within the histology lab as a result of a survey of 12 labs. with annual volumes ranging from 4800 to 77000 cases/year. If your lab is within that range this information could be used by you. I hope this will help you! Ren? J. "Van Eyck, Deb" wrote: Hi all, If anyone has information they would like to share I would appreciate it. We are a hospital lab and we are updating doing competencies. We are interested in Volumes and expectations for embedding, cutting etc, that take into account differeing experience levels, training and quality expectations---does anyone have a good system or have good references or articles that discuss this issue? thanks for your input. Deb This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From JWEEMS <@t> sjha.org Wed Jan 4 18:39:53 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Jan 4 18:40:02 2006 Subject: [Histonet] TouchPreps again Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0130541F@sjhaexc02.sjha.org> How are you all using the new touch prep codes? Do you use only if you are giving a call back result or do you use them for any instead of the 88161? Thanks in advance, and happy new year to all! j Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 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 jdmd77 <@t> hotmail.com Wed Jan 4 21:04:18 2006 From: jdmd77 <@t> hotmail.com (Julia Dahl) Date: Wed Jan 4 21:04:29 2006 Subject: [Histonet] Number of GI biopsy sections on a slide for pediatrichospitals. In-Reply-To: <9E6D52F532809247BDA1783680E92C5658687A@CHWEXC.chwi.chswi.org> Message-ID: Hello Annette - >From the perspective of a gastrointestinal pathologist, the more sections that you can provide for review, the better for the patient - particularly in pediatric patients. At the University of Washington Gastrointestinal Pathology Laboratory, each block would have two complete ribbons of tissue per slide, two slides per block. This would result in about 60 to 120 sections per block for the pathologist to review. The blocks were trimmed very close to the tissue and standard 4 micron sections were ribboned, with ribbons placed about 5 mm from one another on the slides. While this is "overkill" for many standard adult GI cases, such as routine tubular adenomas in adults, the degree of accuracy in inflammatory disorders is impacted substantially by providing additional sections for review. If you would like to contact me directly to discuss, I would be happy to provide some literature about diagnostic accuracy and biopsy handling. Julia Dahl, M.D. Founder, Principal Investigator Mosaic Gastrointestinal Pathology Research Consortium Memphis, TN >From: "Foshey, Annette" >To: >Subject: [Histonet] Number of GI biopsy sections on a slide for >pediatrichospitals. >Date: Wed, 4 Jan 2006 15:50:35 -0600 >MIME-Version: 1.0 >Received: from swlx162.swmed.edu ([199.165.152.162]) by >bay0-mc7-f15.bay0.hotmail.com with Microsoft SMTPSVC(6.0.3790.211); Wed, 4 >Jan 2006 13:55:11 -0800 >Received: from localhost ([127.0.0.1] helo=swlx162.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34)id 1EuGXA-00045Z-1N; Wed, 04 Jan >2006 15:51:09 -0600 >Received: from [199.165.152.166] (helo=swlx166.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34) id 1EuGWt-00045Q-81for >histonet@lists.utsouthwestern.edu; Wed, 04 Jan 2006 15:50:52 -0600 >Received: from chwmail.chw.org ([65.90.124.41])by swlx166.swmed.edu with >esmtp (Exim 4.44) id 1EuGWs-0007p4-1Hfor histonet@lists.utsouthwestern.edu; >Wed, 04 Jan 2006 15:50:47 -0600 >Received: from CHWEXC.chwi.chswi.org (Not Verified[10.1.31.111]) >bychwmail.chw.org with NetIQ MailMarshal (v5.5.6.7)id ; Wed, 04 >Jan 2006 15:50:36 -0600 >X-Message-Info: bIUPYgMBwnfomdUbgby+4zoWKj4Dv1y2rR7pbOUJwfU= >X-MimeOLE: Produced By Microsoft Exchange V6.5 >Content-class: urn:content-classes:message >X-MS-Has-Attach: X-MS-TNEF-Correlator: Thread-Topic: Number of GI biopsy >sections on a slide for pediatric hospitals. >Thread-Index: AcYReOTA825WLiomQ0qPRtInt8A1Rg== >X-Scan-Signature: 74c672b1d43a10f79f2b0e40e5db1c22 >X-Content-Filtered-By: Mailman/MimeDel 2.1.5 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >Precedence: list >List-Id: For the exchange of information pertaining to histotechnology >andrelated fields >List-Unsubscribe: >, > >List-Archive: >List-Post: >List-Help: >List-Subscribe: >, >Errors-To: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: ab3f4c55490e4d672805895add812bc5 >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: X-Spam-Status: No, hits=0.0 required=5.5 tests=none >autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >Return-Path: histonet-bounces@lists.utsouthwestern.edu >X-OriginalArrivalTime: 04 Jan 2006 21:55:12.0092 (UTC) >FILETIME=[896C29C0:01C61179] > > >I am inquiring what the "standard" is for the number of GI biopsy >sections that are routinely put on slides for review. We currently cut >two slides with two levels on each slide. The total number of sections >is between 6-8 sections on each slide. > >Thanks, >Annette Foshey, HT >Team Leader in Histology >Children's Hospital of Wisconsin > > >************************** >Children's Hospital and Health System recognizes that unencrypted e-mail is >insecure 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 _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From Eric <@t> ategra.com Wed Jan 4 20:47:47 2006 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Wed Jan 4 21:34:07 2006 Subject: [Histonet] Immediate Job Opportunities For HistoTechs Nationwide (perm and contract) Message-ID: Histonetters - I have temporary travel assignments and permanent HistoTech openings available for you right now throughout the country! I also have a temporary assignments. If you are currently available, please contact me ASAP at (800) 466-9919, ext. 223. Our clients are now hiring and need to fill these positions ASAP. Thanks for your time/attention... Eric Dye (800) 466-9919, ext. 223 PS - If you know anyone else who is currently looking, I would appreciate it if you could email me there phone number/email address. (Your friend would also appreciate it too!) PSS - If YOU are interested in any of these opportunities, please call me ASAP @ (800) 466 9919 x223. To speed things up, please send me a copy of your resume, (if you haven't already done so) - There is absolutely N O C O S T to you whatsoever! Our services are entirely paid for by the hiring companies. Eric Dye (800) 466-9919, ext. 223 Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792 (800)466-9919 ext 231 FAX: (407) 671-6075 eric@ategra.com To Learn More About Ategra: http://www.ategra.com -------------------------------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again -------------------------------------------------------------------------------------- -------------------------------------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu -------------------------------------------------------------------------------------- From L.Driessen <@t> orthop.umcn.nl Thu Jan 5 01:58:05 2006 From: L.Driessen <@t> orthop.umcn.nl (L.Driessen@orthop.umcn.nl) Date: Thu Jan 5 01:58:23 2006 Subject: [Histonet] Re: Plastic sectioning Message-ID: <2E2AC813A84E054C8E8E572131082BE2145520@umcnet14.umcn.nl> Dear Sunny, First about the knife marks. To me it seems that you are using a blunt knife. Try a new/sharp one. About the shattering: I don't know how youre blocks look like. Is the GMA well impregnated into the brain-tissue, for else this could explain the problem. If this is not the case (I don't think, else you would have mentioned it) then perhaps you can try to use some sort of tape. Stick it onto the block and then cut you're slice. I've tried it and it worked in my case. The only problem then is how to remove the tape without damaging the slice (unless you stain the slices when still sticked to the tape). There are also commercial tapes specially for this technique (see for instance this link: http://www.alphelys.com/site/us/pHGP_CoupesParaffine.htm). Best Regards, l?on From c.m.vanderloos <@t> amc.uva.nl Thu Jan 5 04:41:48 2006 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Thu Jan 5 04:42:01 2006 Subject: [Histonet] RE: optimal thickness for cutting of IHC sections Message-ID: <1d7b4e1d1bca.1d1bca1d7b4e@amc.uva.nl>

Dear Paul,

I totally agree what you wrote about IHC: it's just a

Long time ago we had to prove that our (prehistoric) attempt of qu due to different thi on the staining i that thicker sections staining than thinner sections. style="FONT-SIZE: 10pt; COLOR: black; FONT-FAM Arial">CM van der Lo os, MMH Marijianowski and AE Becker: Quantification in immunohistoche collagen types I and I 347-354< style="FONT-SIZE: 10pt; COLOR: black; FONT-FAM Arial">Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M Nether "urn:schemas-microsoft />

 

Date: Tue, 3 Jan 2006 13:09:20 -0500
From: "Mo <PMonfils@Lifespan.org>
Subject [Histonet] optimal thickness for cutting of IHC sections< BR>To: 'histonet@lists.utsouthwestern.edu'
because anti penetrate tissue very of the section, you are exposed
surface of the tissue, perhaps microns or so.  We have
verified this b microscopy.  This is quite different from standard< BR>histochemical procedures where a thicker section results in a more i the tissue through.

From TMcNemar <@t> lmhealth.org Thu Jan 5 06:26:12 2006 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Thu Jan 5 06:31:22 2006 Subject: [Histonet] TouchPreps again Message-ID: <6CD94D97ED7D924BA5C2B588FA95282139682C@nt_exchange.lmhealth.org> Our understanding is that they are only to be used as part of (or instead of) a frozen section. We are using them for sentinel node touch preps. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Wednesday, January 04, 2006 7:40 PM To: Histonet Subject: [Histonet] TouchPreps again How are you all using the new touch prep codes? Do you use only if you are giving a call back result or do you use them for any instead of the 88161? Thanks in advance, and happy new year to all! j Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 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 deb.vaneyck <@t> phci.org Thu Jan 5 09:18:01 2006 From: deb.vaneyck <@t> phci.org (Van Eyck, Deb) Date: Thu Jan 5 09:18:16 2006 Subject: [Histonet] Happy New Years and a question? Message-ID: Hi all, I would appreciate any input or sharing of resources, procedures, articles, etc. We are looking at our competancies and workload recording process. We are a hospital lab and I would like to hear from you if you have a good policy for workload recording, expectations, etc for hsto staff, all experience levels---aimed at also quality end products. thanks-Deb This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. From jackdodo <@t> msn.com Thu Jan 5 09:56:06 2006 From: jackdodo <@t> msn.com (WAYNE HOLLAND) Date: Thu Jan 5 09:56:16 2006 Subject: [Histonet] Human pro-collagen 1 Message-ID: Has anyone used this antibody in a pre-diluted form? The only place I have located this antibody is from Chemicon (non-diluted). Thanks From joycefr <@t> frontiernet.net Thu Jan 5 10:19:40 2006 From: joycefr <@t> frontiernet.net (Joyce Friedland) Date: Thu Jan 5 10:01:51 2006 Subject: [Histonet] NYS Certification Requirements Message-ID: <277a2d600ecceecacc74eb9f06c15f12@frontiernet.net> Hi All You NYS Histotechs, I was just told that there will be, in the near future, a requirement that all histotechs in NYS be certified. Can anyone give me information on this. Will there be a "grandfathering" provision? When is this going into effect? Thanks, Joyce From nadams <@t> CapeCodHealth.org Thu Jan 5 10:13:30 2006 From: nadams <@t> CapeCodHealth.org (Adams, Nancy) Date: Thu Jan 5 10:28:48 2006 Subject: [Histonet] document for recording special stain results for quality control Message-ID: <17A1862099540D458C8FE9380C2BC461C09316@fh2xmail.fhdomain1.capecodhealth.org> Does anyone have a good document for recording this information that they'd be willing to share? I'm in the process of getting ready for a CAP inspection. Thanks Nancy Rutledge Falmouth Hospital Falmouth, MA ************************************************************ 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 contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ From BlazekL <@t> childrensdayton.org Thu Jan 5 10:59:35 2006 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Thu Jan 5 11:01:52 2006 Subject: [Histonet] document for recording special stain results for quality control Message-ID: Nancy, We document the special stain results and the control results on the pathology report. We have a protocol that states all stains will be reviewed by the tech doing the stain before they are given to the pathologist so there should never be a poor review on the pathology report. We have a form that the pathologist can fill out and return to the department supervisor if there is a problem with the stain they receive. We also put a note on the case in the computer to document why a case may have taken longer than expected to sign out due to a problem with a special stain. If I have be of any more help let me know. Linda Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org From rjbuesa <@t> yahoo.com Thu Jan 5 12:00:23 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Jan 5 12:00:31 2006 Subject: [Histonet] document for recording special stain results for quality control In-Reply-To: <17A1862099540D458C8FE9380C2BC461C09316@fh2xmail.fhdomain1.capecodhealth.org> Message-ID: <20060105180023.17687.qmail@web61221.mail.yahoo.com> Hi Nancy: We used to "batch" special stains (1 positive control for slides with the same procedure, either IHC, HC or even Her-2new), so we had a "common" form where we used to write the date, the test we batched, and the cases numbers included in the batch. We checked if the control reacted as expected, attached the form and the control to the batched slides and gave them to the pathologist. The form was originally signed by the histotech who did the test and it was also signed by the pathologist. All the forms were filed by the type of test (Steiner, IHCs on the day, PAS, etc.) and were available for inspection. In our last CAP inspection we were told that this was a good procedure. I hope this will help you! Ren? J. "Adams, Nancy" wrote: Does anyone have a good document for recording this information that they'd be willing to share? I'm in the process of getting ready for a CAP inspection. Thanks Nancy Rutledge Falmouth Hospital Falmouth, MA ************************************************************ 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 contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less From lldewe <@t> ucdavis.edu Thu Jan 5 12:11:58 2006 From: lldewe <@t> ucdavis.edu (Loralei Dewe) Date: Thu Jan 5 12:12:07 2006 Subject: [Histonet] Smear preps In-Reply-To: References: Message-ID: I need to do some smears from frozen tissue. I've never done them before but they don't sound all that tough. Anyone got a good protocol to share with me? Cheers, Loralei Dewe UC Davis Vet Hospital From BlazekL <@t> childrensdayton.org Thu Jan 5 12:12:52 2006 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Thu Jan 5 12:15:15 2006 Subject: [Histonet] document for recording special stain results for quality control Message-ID: I forgot to add something to my previous note. We run batch controls also as Rene does but we have a control label ie: GMS #1, IRON #1 etc. The control number is also dictated on the pathology report. That way we can always go back to the correct control slide - Linda Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Adams, Nancy" 01/05/2006 11:13 AM >>> Does anyone have a good document for recording this information that they'd be willing to share? I'm in the process of getting ready for a CAP inspection. Thanks Nancy Rutledge Falmouth Hospital Falmouth, MA ************************************************************ 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 contact the system administrator for Cape Cod Healthcare. Administrator@CapeCodHealth.org ************************************************************ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Jan 5 14:22:24 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Jan 5 14:22:40 2006 Subject: [Histonet] Re:Plastic sectioning In-Reply-To: <2E2AC813A84E054C8E8E572131082BE2145520@umcnet14.umcn.nl> References: <2E2AC813A84E054C8E8E572131082BE2145520@umcnet14.umcn.nl> Message-ID: <6.0.0.22.1.20060105130946.01b60008@gemini.msu.montana.edu> Sunny, You have a tough problem here. 30 um thick GMA sections may be too thick for this embedding media. GMA plastic was initially designed for doing THIN sections at 1 to 3 micrometers. It tends to be brittle when cutting thick sections and one might have to use a very sharp tungsten carbide tipped knife in order to get any thick section off a GMA block. One could try to adjust the plasticizer by increasing its concentration to make the GMA more pliable. The knife, no matter how sharp, may not actually slice through the plastic, but force its way through in such a way that the plastic shatters i.e. a shearing effect and not really have a clean cut. Patsy Ruegg published an inhouse method for GMA, rather than buy kits - that way she could adjust the ingredients accordingly for softer or harder GMA. If the tissues are thicker than 1 to 2 mm when processing and embedding, infiltration may be a problem unless extended time is used and done at 4C ( refrigerator) and polymerization might have to controlled with cold temperatures in order to slow it down for continuous or steady progression of polymerizing. That can be done at 4C also. At 12:58 AM 1/5/2006, you wrote: >Dear Sunny, > >First about the knife marks. To me it seems that you are using a blunt >knife. Try a new/sharp one. >About the shattering: I don't know how youre blocks look like. Is the GMA >well impregnated into the brain-tissue, for else this could explain the >problem. If this is not the case (I don't think, else you would have >mentioned it) then perhaps you can try to use some sort of tape. Stick it >onto the block and then cut you're slice. I've tried it and it worked in >my case. The only problem then is how to remove the tape without damaging >the slice (unless you stain the slices when still sticked to the tape). >There are also commercial tapes specially for this technique (see for >instance this link: http://www.alphelys.com/site/us/pHGP_CoupesParaffine.htm). Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From david.kinsley <@t> spcorp.com Thu Jan 5 15:32:50 2006 From: david.kinsley <@t> spcorp.com (Kinsley, David) Date: Thu Jan 5 15:33:17 2006 Subject: [Histonet] Happy New Year's and competancy/volume questions Message-ID: Hi all, It seems to me that the competencies below are for human tissues in a hospital/clinical setting. I was wondering if there are competency/productivity standards for research histology on animal tissues particularly mouse and rat species, paraffin and frozen. We are looking to establish some base guidelines for our lab. Any suggestions are appreciated. thanks Dave -----Original Message----- From: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: Wednesday, January 04, 2006 6:33 PM To: Van Eyck, Deb; ' histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] Happy New Year's and competancy/volume questions Hi Deb: In 3 labs I wrote competencies for the expectation for sectoning was set at 25 blocks/hour and for embedding at 60 blocks/hour. Each histotech was evaluated on that basis and their individual averages were rated (as percentages) against the expectation. Those with averages above expectations were evaluated better (and received salary compensations accordingly). Nobody was considered "proficient" until their individual average reached the expectation consistently. For time utilization (ratio between time with recorded tasks versus logged time) the expectation was 75 % (i.e. 6 h with logged tasks agains 8 h/day per shift). In the Dec./04 issue of the Journal of Histotechnology there are averages for every imaginable task within the histology lab as a result of a survey of 12 labs. with annual volumes ranging from 4800 to 77000 cases/year. If your lab is within that range this information could be used by you. I hope this will help you! Ren? J. "Van Eyck, Deb" wrote: Hi all, If anyone has information they would like to share I would appreciate it. We are a hospital lab and we are updating doing competencies. We are interested in Volumes and expectations for embedding, cutting etc, that take into account differeing experience levels, training and quality expectations---does anyone have a good system or have good references or articles that discuss this issue? thanks for your input. Deb This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From Tom_Wells <@t> bcit.ca Thu Jan 5 16:06:00 2006 From: Tom_Wells <@t> bcit.ca (Tom Wells) Date: Thu Jan 5 16:06:10 2006 Subject: [Histonet] gross room Message-ID: I am going to be discussing the gross room and prosecting with my medical laboratory technology students soon. This is an area that seems to be rather poorly documented. Do any of you have gross room manuals that you wouldn't mind sharing with me? Are there any web sites which discuss grossing for new residents? Are there any pathology sites that have established standards for grossing etc. Any help would be gratefully appreciated. Thanks. Tom From gentras <@t> vetmed.auburn.edu Thu Jan 5 16:36:05 2006 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Thu Jan 5 16:36:22 2006 Subject: [Histonet] 50 micron mouse brain Message-ID: <6.0.1.1.0.20060105163140.01acf798@mailhost.vetmed.auburn.edu> Hello, will someone please give me pointers on obtaining good 50u sections of PFA drop fixed and/or immersion fixed coronal paraffin sections of mouse brain? As I section they tend to roll up in tight rolls. I've tried sectioning at room temp. vs. chilled sections. Any assistance provided will be greatly appreciated. Thanks, Atoska From lenaspencer <@t> insightbb.com Thu Jan 5 17:55:31 2006 From: lenaspencer <@t> insightbb.com (lena spencer) Date: Thu Jan 5 17:50:13 2006 Subject: [Histonet] Job Opportunity Message-ID: <005401c61253$8311b880$3379df0c@org.insightbb.com> Electron Microscopy Job Opportunity Full Time/Day Shift some histology duties required Norton Healthcare Louisville, Kentucky Contact: Mary Beth Knight, MT(ASCP)HTL marybeth.knight@nortonhealthcare.org 502-629-7884 From jnocito <@t> satx.rr.com Thu Jan 5 21:35:07 2006 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Thu Jan 5 21:35:22 2006 Subject: [Histonet] Number of GI biopsy sections on a slide for pediatrichospitals. References: <9E6D52F532809247BDA1783680E92C5658687A@CHWEXC.chwi.chswi.org> Message-ID: <007901c61272$30e4f1c0$e8bd0b43@yourxhtr8hvc4p> Annette, we cut 3 levels on one slide with 2 sections per level Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Foshey, Annette" To: Sent: Wednesday, January 04, 2006 3:50 PM Subject: [Histonet] Number of GI biopsy sections on a slide for pediatrichospitals. I am inquiring what the "standard" is for the number of GI biopsy sections that are routinely put on slides for review. We currently cut two slides with two levels on each slide. The total number of sections is between 6-8 sections on each slide. Thanks, Annette Foshey, HT Team Leader in Histology Children's Hospital of Wisconsin ************************** Children's Hospital and Health System recognizes that unencrypted e-mail is insecure 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 -- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.1.371 / Virus Database: 267.14.13/221 - Release Date: 1/4/2006 From christy0424 <@t> gmail.com Fri Jan 6 00:35:02 2006 From: christy0424 <@t> gmail.com (Kuo Christy) Date: Fri Jan 6 00:35:11 2006 Subject: [Histonet] Re: CD4 and CD8 in mouse / human FFPE In-Reply-To: <5.1.0.14.0.20051219090417.00c4a238@mail.jcu.edu.au> References: <43A5CB51.20138.3A8527E7@localhost> <5.1.0.14.0.20051219090417.00c4a238@mail.jcu.edu.au> Message-ID: <5e4834130601052235w191fb3f9g206edcd8ac567a9e@mail.gmail.com> Hi Reinhard, Could I also ask for a copy of your protocol, cuz the results from anti-CD4(Novocvastra) seems to be very unspecific on human tissue FFPE sections and I don't know the reason. Thank you for your help! Best wishes, Christy christy0424@gmail.com On 12/19/05, Laurie Reilly wrote: > Reinhard, > Could I please have a copy of your protocol, we have had lots of trouble > trying to stain with > CD4 and CD8 in bovine tissues. > Any advice you can give would be most welcome, particularly about antigen > retrieval. > > Hope everyone has a happy Christmas, > Regards, > Laurie. > > At 08:49 PM 18/12/2005 +0100, you wrote: > >Hi Patsy, > >we do a lot of CD4 and CD8 in human FFPE tissues, even in BM after > >decalification. > >It works very good and is daily routine in our lab. If you need advice, > >give me a mail. > >We now got some hints how both markers could work in mouse tissue, too. If > we > >succeed, we'll let you know. > > > >best regards > >Reinhard. > > > > > > > >PD Dr. med. Reinhard von Wasielewski > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Mr.Laurie Reilly Ph 07 4781 > 4468 > School of Veterinary & Biomedical Science Fax 07 4779 1526 > James Cook University > Townsville Qld. > 4811 laurie.reilly@jcu.edu.au > > Australia. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From LJApuzzio <@t> msn.com Fri Jan 6 04:54:04 2006 From: LJApuzzio <@t> msn.com (Louis Apuzzio) Date: Fri Jan 6 04:54:15 2006 Subject: [Histonet] RE: Message-ID: Anyone in search of a Histotech, ( HT eligible) and also a Cytotech, ( CT(ASCP), for small private laboratory and or hospital, please contact: ljapuzzio@msn.com Thank you ! From c.m.vanderloos <@t> amc.uva.nl Fri Jan 6 05:26:41 2006 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Jan 6 05:26:54 2006 Subject: [Histonet] RE: optimal thickness for cutting of IHC sections Message-ID: <1d9a431dc72b.1dc72b1d9a43@amc.uva.nl> Sorry guys my message yesterday was somewhat distorted. Here it is again... Dear Paul, I totally agree what you wrote about IHC: it's just a surface event. Long time ago we had to prove that our (prehistoric) attempt of quantify due to microtome different thickness and gue on the staining intensity. The only thicker sections showed a higher non-specific b than thinner sections. See: CM van der Loos, MMH Marijianowski and AE Becker: Quantification in immu collagen types I Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Tue, 3 Jan 2006 13:09:20 -0500 From: "Monfils, Paul" 6 && arg.substring(0, 7) == 'bgcolor') color(arg.substring(8, arg.length)) f = eval('parent.' + self.name + 'Init') if (f) f() } // Note: the tag is defined in main.js and written out when a given is created. /**Enable backspace for text editing*/ if (NN>=5){ setBkspEvtHndlr("input");//specifically for text elements. checkboxes/r setBkspEvtHndlr("textarea"); setBkspEvtHndlr("browse"); } References 1. 3D"javascript:main.compose('new','t='hi From froyer <@t> bitstream.net Fri Jan 6 08:26:31 2006 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Jan 6 08:26:46 2006 Subject: [Histonet] Looking for a home... Message-ID: <002001c612cd$30f0a230$3900a8c0@fords> A friend has asked me to try and find a home for her retired TissueTek VIP (K-series) Tissue Processors. These would make a great back-up unit or a basic unit in a start-up lab. Contact me Off-List for details. ~ Ford Ford M. Royer, MT(ASCP) Minnesota Medical Specialists, Inc. Golden Valley, MN 55427-3601 763-542-8725 phone 888-790-9686 toll free 763-546-4830 fax email From dfleisch <@t> bidneedham.org Fri Jan 6 08:46:22 2006 From: dfleisch <@t> bidneedham.org (dfleisch@bidneedham.org) Date: Fri Jan 6 08:46:36 2006 Subject: [Histonet] slide dryer Message-ID: I am looking for a small slide dryer. Would greatly appreciate any suggestions. Deborah S. Fleischhacker, M.D. Pathologist Beth Israel Deaconess Hospital Needham 148 Chestnut Street Needham, MA 02492 Phone: (781) 453-3087 Fax: (781) 453-3097 From RRA <@t> Stowers-Institute.org Fri Jan 6 10:19:39 2006 From: RRA <@t> Stowers-Institute.org (Allen, Rhonda) Date: Fri Jan 6 10:20:04 2006 Subject: [Histonet] Processing/embedding question - your opinion Message-ID: Have done it on many occasions and never had it be a problem. Rhonda Allen Stowers Institute 1000 E. 50th street Kansas City, Missouri 64110 rra@stowers-institute.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of foley1 Sent: Thursday, December 29, 2005 11:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Processing/embedding question - your opinion Does anyone routinely allow for the hot wax to drain off multiple cassettes of processed tissue and be held at room temperature for multiple days (6 days) before embedding? What would the consequences be to morphology, possible immunohistochemistry and molecular (DNA) studies? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mprice26 <@t> juno.com Fri Jan 6 10:49:27 2006 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Jan 6 10:51:21 2006 Subject: [Histonet] RE: Price quotes on refurbished lab equipment/ Start Up Lab Message-ID: <20060106.085006.9834.259217@webmail48.nyc.untd.com> Histonetters, First of all, Happy New Year to everyone in histoland. I am looking for companies that can sell all the equipment (refurbished) needed for a start up lab. I would like a package deal, everything included, IE Processor, embedding center, microtome, coverslipper, & waterbath all together. Vendors are welcome to respond. Thank you. Marsha Price From rjbuesa <@t> yahoo.com Fri Jan 6 10:57:38 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Jan 6 10:57:45 2006 Subject: [Histonet] Processing/embedding question - your opinion In-Reply-To: Message-ID: <20060106165738.73387.qmail@web61219.mail.yahoo.com> Hi Foley: I have seen that done in several places. I have also seen all the cassettes wrapped in aluminum foil and kept that way (like a pizza left-over). I cannot tell you what would be the consequences for the morphology or future tests. Theoretically speaking probably this practice will not affect because the tissue itself and all its components are supposedly embedded in paraffin that will just solidify. I personally do not like this to be done. For me, personally, it indicates laziness, indolence and even "disrespect" for the tissue sample. Once I was confronted with the need of keeping processed cassettes in a secure way before casting the blocks. My solution was to put all the cassettes in a shallow plastic container, place all the blocks in it, add melted paraffin and prepare one single block, as large as the container. When I was able to prepare the blocks individually, I melted the paraffin and prepared the blocks. Hope this will help you! Ren? J. foley1 wrote: Does anyone routinely allow for the hot wax to drain off multiple cassettes of processed tissue and be held at room temperature for multiple days (6 days) before embedding? What would the consequences be to morphology, possible immunohistochemistry and molecular (DNA) studies? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less From lharris <@t> samhealth.org Fri Jan 6 10:57:49 2006 From: lharris <@t> samhealth.org (lharris@samhealth.org) Date: Fri Jan 6 10:58:02 2006 Subject: [Histonet] Microwave slide drying vs. conventional slide drying Message-ID: <5B3B26C4B71D5E469892D1860ABE10EA7F1E7F@SHSEXVS01.int.samhealth.net> We are having a discussion about slide drying at out facility. How many of you out there use a microwave to dry your H&E slides before staining versus using a conventional 70 degree Celsius hot air slide drying oven for 20 minutes? I personally against microwave slide drying but the doctors like it because it takes less time (3 min. 30 sec. on medium setting). Thanks for your response. Lori A. Harris, HT (ASCP) Histology Section Leader GSRMC - Pathology 3600 NW Samaritan Drive Corvallis, OR 97330 1-541-768-6078 lharris@samhealth.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 sjchtascp <@t> yahoo.com Fri Jan 6 11:23:14 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Jan 6 11:23:22 2006 Subject: [Histonet] sucrose liver Message-ID: <20060106172314.29262.qmail@web90204.mail.scd.yahoo.com> Good morning everyone, If anyone out there cryo-sections sucrose infiltrated fixed liver I'd like to know what temp there are sectioning at best. The mouse liver is fixed at 4C in formalin/PBS for 6 hours the 25-30% sucrose overnight also at 4C. I thought sucrose infiltrated tissue should section closer to -25C. Our chatter at anything above -18C. Steve --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less From hborgeri <@t> wfubmc.edu Fri Jan 6 11:23:26 2006 From: hborgeri <@t> wfubmc.edu (Hermina Borgerink) Date: Fri Jan 6 11:23:35 2006 Subject: [Histonet] HTL certification Message-ID: <9AEEF1FB6254224AA355ED285F84916515A37AA7@EXCHVS2.medctr.ad.wfubmc.edu> Two of my techs will be taking the HTL certification this year. I checked the requirements on the ASCP Board of Registry home page and found that they needed experience in the following areas: fixation, processing, microtomy, and routine as well as special staining. I just wanted to double check with those of you who have recently taken their Board Certifications that EM and ISH experience are no longer required. When I did my HTL certification there still were questions pertaining to both topics, and I just want to make sure they won't be faced with questions they are not trained in. Any response would be greatly appreciated and I thank you in advance. Hermina Hermina M. Borgerink, BA, HT(ASCP)HTL, QIHC Wake Forest University Health Sciences Department of Pathology Medical Center Blvd. Winston-Salem, NC 27157 Tel. (336) 716-1538 Fax. (336) 716-1515 e-mail hborgeri@wfubmc.edu From bwhitaker <@t> brownpathology.com Fri Jan 6 11:32:30 2006 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Jan 6 11:32:37 2006 Subject: [Histonet] Processing/embedding question - your opinion In-Reply-To: <20060106165738.73387.qmail@web61219.mail.yahoo.com> Message-ID: <000001c612e7$2be04060$3601a8c0@brownpathology.net> Rene, What is "lazy and indolent" about saving processed tissue without embedding it? I frequently collect many tissues for embedding in multi-tissue blocks, and I see no point in embedding the tissue temporarily while I continue to collect additional material. It makes no difference if the paraffin coating is minimal or a full block. You can also store a lot more control tissue in a small area if it is maintained in unembedded cassettes. One cassette can hold several blocks worth of material. Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, January 06, 2006 10:58 AM To: foley1; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Processing/embedding question - your opinion Hi Foley: I have seen that done in several places. I have also seen all the cassettes wrapped in aluminum foil and kept that way (like a pizza left-over). I cannot tell you what would be the consequences for the morphology or future tests. Theoretically speaking probably this practice will not affect because the tissue itself and all its components are supposedly embedded in paraffin that will just solidify. I personally do not like this to be done. For me, personally, it indicates laziness, indolence and even "disrespect" for the tissue sample. Once I was confronted with the need of keeping processed cassettes in a secure way before casting the blocks. My solution was to put all the cassettes in a shallow plastic container, place all the blocks in it, add melted paraffin and prepare one single block, as large as the container. When I was able to prepare the blocks individually, I melted the paraffin and prepared the blocks. Hope this will help you! Ren? J. foley1 wrote: Does anyone routinely allow for the hot wax to drain off multiple cassettes of processed tissue and be held at room temperature for multiple days (6 days) before embedding? What would the consequences be to morphology, possible immunohistochemistry and molecular (DNA) studies? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Fri Jan 6 11:38:07 2006 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Jan 6 11:38:22 2006 Subject: [Histonet] Microwave slide drying vs. conventional slide drying Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628005653929@EXCHANGE1.huntingtonhospital.com> Lori, We don't use either. We simply put our drained slides on the hot plate for about 5 minutes. In previous jobs, we always used an oven, and when I came here I was really nervous about just using the hot plate - but it works! Laurie Colbert Huntington Hospital Pasadena, CA -----Original Message----- From: lharris@samhealth.org [mailto:lharris@samhealth.org] Sent: Friday, January 06, 2006 8:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave slide drying vs. conventional slide drying We are having a discussion about slide drying at out facility. How many of you out there use a microwave to dry your H&E slides before staining versus using a conventional 70 degree Celsius hot air slide drying oven for 20 minutes? I personally against microwave slide drying but the doctors like it because it takes less time (3 min. 30 sec. on medium setting). Thanks for your response. Lori A. Harris, HT (ASCP) Histology Section Leader GSRMC - Pathology 3600 NW Samaritan Drive Corvallis, OR 97330 1-541-768-6078 lharris@samhealth.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 From rjbuesa <@t> yahoo.com Fri Jan 6 11:43:24 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Jan 6 11:43:33 2006 Subject: [Histonet] Processing/embedding question - your opinion In-Reply-To: <000001c612e7$2be04060$3601a8c0@brownpathology.net> Message-ID: <20060106174324.46935.qmail@web61222.mail.yahoo.com> Bonnie: That is my personal and very particular opinion, and I think I am entitled to that, in the same way that you have expressed yours. I believe that this is covered by the freedom of expression, is it not? Ren? J. Bonnie Whitaker wrote: Rene, What is "lazy and indolent" about saving processed tissue without embedding it? I frequently collect many tissues for embedding in multi-tissue blocks, and I see no point in embedding the tissue temporarily while I continue to collect additional material. It makes no difference if the paraffin coating is minimal or a full block. You can also store a lot more control tissue in a small area if it is maintained in unembedded cassettes. One cassette can hold several blocks worth of material. Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, January 06, 2006 10:58 AM To: foley1; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Processing/embedding question - your opinion Hi Foley: I have seen that done in several places. I have also seen all the cassettes wrapped in aluminum foil and kept that way (like a pizza left-over). I cannot tell you what would be the consequences for the morphology or future tests. Theoretically speaking probably this practice will not affect because the tissue itself and all its components are supposedly embedded in paraffin that will just solidify. I personally do not like this to be done. For me, personally, it indicates laziness, indolence and even "disrespect" for the tissue sample. Once I was confronted with the need of keeping processed cassettes in a secure way before casting the blocks. My solution was to put all the cassettes in a shallow plastic container, place all the blocks in it, add melted paraffin and prepare one single block, as large as the container. When I was able to prepare the blocks individually, I melted the paraffin and prepared the blocks. Hope this will help you! Ren? J. foley1 wrote: Does anyone routinely allow for the hot wax to drain off multiple cassettes of processed tissue and be held at room temperature for multiple days (6 days) before embedding? What would the consequences be to morphology, possible immunohistochemistry and molecular (DNA) studies? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less From RRA <@t> Stowers-Institute.org Fri Jan 6 11:51:03 2006 From: RRA <@t> Stowers-Institute.org (Allen, Rhonda) Date: Fri Jan 6 11:51:38 2006 Subject: [Histonet] Processing/embedding question - your opinion Message-ID: Renee, It may be your opinion, but to those of us who have done it out of necessity, not being "lazy and indolent", it sounds like you are trying to make us feel "lazy and indolent". Rhonda Allen BA HT(ASCP)HTL, QIHC Histotechnology Specialist II Stowers Institute 1000 E. 50th Street Kansas City, MO 64110 816-926-4305 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, January 06, 2006 11:43 AM To: Bonnie Whitaker; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Processing/embedding question - your opinion Bonnie: That is my personal and very particular opinion, and I think I am entitled to that, in the same way that you have expressed yours. I believe that this is covered by the freedom of expression, is it not? Ren? J. Bonnie Whitaker wrote: Rene, What is "lazy and indolent" about saving processed tissue without embedding it? I frequently collect many tissues for embedding in multi-tissue blocks, and I see no point in embedding the tissue temporarily while I continue to collect additional material. It makes no difference if the paraffin coating is minimal or a full block. You can also store a lot more control tissue in a small area if it is maintained in unembedded cassettes. One cassette can hold several blocks worth of material. Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, January 06, 2006 10:58 AM To: foley1; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Processing/embedding question - your opinion Hi Foley: I have seen that done in several places. I have also seen all the cassettes wrapped in aluminum foil and kept that way (like a pizza left-over). I cannot tell you what would be the consequences for the morphology or future tests. Theoretically speaking probably this practice will not affect because the tissue itself and all its components are supposedly embedded in paraffin that will just solidify. I personally do not like this to be done. For me, personally, it indicates laziness, indolence and even "disrespect" for the tissue sample. Once I was confronted with the need of keeping processed cassettes in a secure way before casting the blocks. My solution was to put all the cassettes in a shallow plastic container, place all the blocks in it, add melted paraffin and prepare one single block, as large as the container. When I was able to prepare the blocks individually, I melted the paraffin and prepared the blocks. Hope this will help you! Ren? J. foley1 wrote: Does anyone routinely allow for the hot wax to drain off multiple cassettes of processed tissue and be held at room temperature for multiple days (6 days) before embedding? What would the consequences be to morphology, possible immunohistochemistry and molecular (DNA) studies? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sandra.Harrison3 <@t> va.gov Fri Jan 6 11:53:08 2006 From: Sandra.Harrison3 <@t> va.gov (Harrison, Sandra C.) Date: Fri Jan 6 11:53:19 2006 Subject: [Histonet] microwave tissue processing Message-ID: <736E8889E98B8F4FBBD29FDFEC8074BA49BF79@VHAV23MSGA2.v23.med.va.gov> Hi, I'm looking into microwave tissue processors. If you are using one, please tell me the brand and whether you are pleased with the results. Do you find the instrument reliable? Thanks, Histonetters. Sandy From dusko.trajkovic <@t> pfizer.com Fri Jan 6 11:54:58 2006 From: dusko.trajkovic <@t> pfizer.com (Trajkovic, Dusko) Date: Fri Jan 6 11:55:14 2006 Subject: [Histonet] Processing/embedding question - your opinion Message-ID: <3AD0BD3142459B4E9B12CBEAFF2B89B2D896AB@lajamrexm01.amer.pfizer.com> I have yet to see any published information stating that un-embedded tissue left to solidify, is damaging in any way or form. My colleagues an I in research, have done it numerous times, for various reasons, non were for being lazy. Dusko Trajkovic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, January 06, 2006 9:43 AM To: Bonnie Whitaker; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Processing/embedding question - your opinion Bonnie: That is my personal and very particular opinion, and I think I am entitled to that, in the same way that you have expressed yours. I believe that this is covered by the freedom of expression, is it not? Ren? J. Bonnie Whitaker wrote: Rene, What is "lazy and indolent" about saving processed tissue without embedding it? I frequently collect many tissues for embedding in multi-tissue blocks, and I see no point in embedding the tissue temporarily while I continue to collect additional material. It makes no difference if the paraffin coating is minimal or a full block. You can also store a lot more control tissue in a small area if it is maintained in unembedded cassettes. One cassette can hold several blocks worth of material. Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, January 06, 2006 10:58 AM To: foley1; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Processing/embedding question - your opinion Hi Foley: I have seen that done in several places. I have also seen all the cassettes wrapped in aluminum foil and kept that way (like a pizza left-over). I cannot tell you what would be the consequences for the morphology or future tests. Theoretically speaking probably this practice will not affect because the tissue itself and all its components are supposedly embedded in paraffin that will just solidify. I personally do not like this to be done. For me, personally, it indicates laziness, indolence and even "disrespect" for the tissue sample. Once I was confronted with the need of keeping processed cassettes in a secure way before casting the blocks. My solution was to put all the cassettes in a shallow plastic container, place all the blocks in it, add melted paraffin and prepare one single block, as large as the container. When I was able to prepare the blocks individually, I melted the paraffin and prepared the blocks. Hope this will help you! Ren? J. foley1 wrote: Does anyone routinely allow for the hot wax to drain off multiple cassettes of processed tissue and be held at room temperature for multiple days (6 days) before embedding? What would the consequences be to morphology, possible immunohistochemistry and molecular (DNA) studies? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------- LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From ander093 <@t> tc.umn.edu Fri Jan 6 12:04:12 2006 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Fri Jan 6 12:04:37 2006 Subject: [Histonet] Processing/embedding question - your opinion In-Reply-To: <3AD0BD3142459B4E9B12CBEAFF2B89B2D896AB@lajamrexm01.amer.pf izer.com> References: <3AD0BD3142459B4E9B12CBEAFF2B89B2D896AB@lajamrexm01.amer.pfizer.com> Message-ID: <6.2.3.4.0.20060106120242.03b2f4d0@ander093.email.umn.edu> Why not do a comparison--take two pieces of the same tissue. Process both-embed one immediately and delay the second. Cut, stain and compare morphology. LuAnn At 11:54 AM 1/6/2006, Trajkovic, Dusko wrote: >I have yet to see any published information >stating that un-embedded tissue left to >solidify, is damaging in any way or form. My >colleagues an I in research, have done it >numerous times, for various reasons, non were for being lazy. > >Dusko Trajkovic > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa >Sent: Friday, January 06, 2006 9:43 AM >To: Bonnie Whitaker; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Processing/embedding question - your opinion > >Bonnie: > That is my personal and very particular > opinion, and I think I am entitled to that, > in the same way that you have expressed yours. > I believe that this is covered by the freedom of expression, is it not? > Ren? J. > >Bonnie Whitaker wrote: > Rene, > >What is "lazy and indolent" about saving processed tissue without embedding >it? I frequently collect many tissues for embedding in multi-tissue blocks, >and I see no point in embedding the tissue temporarily while I continue to >collect additional material. It makes no difference if the paraffin coating >is minimal or a full block. You can also store a lot more control tissue in >a small area if it is maintained in unembedded cassettes. One cassette can >hold several blocks worth of material. > >Bonnie Whitaker >Lab Manager >Brown & Associates Medical Laboratories >8076 El Rio >Houston, Texas 77054 >713-741-6677 > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa >Sent: Friday, January 06, 2006 10:58 AM >To: foley1; histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Processing/embedding question - your opinion > > >Hi Foley: >I have seen that done in several places. I have also seen all the >cassettes wrapped in aluminum foil and kept that way (like a pizza >left-over). >I cannot tell you what would be the consequences for the morphology or >future tests. >Theoretically speaking probably this practice will not affect because the >tissue itself and all its components are supposedly embedded in paraffin >that will just solidify. >I personally do not like this to be done. For me, personally, it indicates >laziness, indolence and even "disrespect" for the tissue sample. >Once I was confronted with the need of keeping processed cassettes in a >secure way before casting the blocks. My solution was to put all the >cassettes in a shallow >plastic container, place all the blocks in it, add melted paraffin and >prepare one single block, as large as the container. When I was able to >prepare the blocks individually, I melted the paraffin and prepared the >blocks. >Hope this will help you! >Ren? J. > >foley1 wrote: >Does anyone routinely allow for the hot wax to drain off multiple >cassettes of processed tissue and be held at room temperature for multiple >days (6 >days) before embedding? What would the consequences be to morphology, >possible immunohistochemistry and molecular (DNA) studies? > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > >--------------------------------- >Yahoo! DSL Something to write home about. Just $16.99/mo. or less >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > >--------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >---------------------------------------------------------------------- >LEGAL NOTICE >Unless expressly stated otherwise, this message >is confidential and may be privileged. It is >intended for the addressee(s) only. Access to >this E-mail by anyone else is unauthorized. If >you are not an addressee, any disclosure or >copying of the contents of this E-mail or any >action taken (or not taken) in reliance on it is >unauthorized and may be unlawful. If you are >not an addressee, please inform the sender immediately. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri Jan 6 12:04:19 2006 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Jan 6 12:04:48 2006 Subject: [Histonet] Processing/embedding question - your opinion Message-ID: I agree, Dusko - I think that when you have a couple-three hundred blocks to embed, it's better to let some solidify than to remain in molten paraffin for an additional 6-7 hours. Jackie O' "Trajkovic, Dusko" Sent by: histonet-bounces@lists.utsouthwestern.edu 01/06/2006 11:54 AM To: cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: RE: [Histonet] Processing/embedding question - your opinion I have yet to see any published information stating that un-embedded tissue left to solidify, is damaging in any way or form. My colleagues an I in research, have done it numerous times, for various reasons, non were for being lazy. Dusko Trajkovic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, January 06, 2006 9:43 AM To: Bonnie Whitaker; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Processing/embedding question - your opinion Bonnie: That is my personal and very particular opinion, and I think I am entitled to that, in the same way that you have expressed yours. I believe that this is covered by the freedom of expression, is it not? Ren? J. Bonnie Whitaker wrote: Rene, What is "lazy and indolent" about saving processed tissue without embedding it? I frequently collect many tissues for embedding in multi-tissue blocks, and I see no point in embedding the tissue temporarily while I continue to collect additional material. It makes no difference if the paraffin coating is minimal or a full block. You can also store a lot more control tissue in a small area if it is maintained in unembedded cassettes. One cassette can hold several blocks worth of material. Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, January 06, 2006 10:58 AM To: foley1; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Processing/embedding question - your opinion Hi Foley: I have seen that done in several places. I have also seen all the cassettes wrapped in aluminum foil and kept that way (like a pizza left-over). I cannot tell you what would be the consequences for the morphology or future tests. Theoretically speaking probably this practice will not affect because the tissue itself and all its components are supposedly embedded in paraffin that will just solidify. I personally do not like this to be done. For me, personally, it indicates laziness, indolence and even "disrespect" for the tissue sample. Once I was confronted with the need of keeping processed cassettes in a secure way before casting the blocks. My solution was to put all the cassettes in a shallow plastic container, place all the blocks in it, add melted paraffin and prepare one single block, as large as the container. When I was able to prepare the blocks individually, I melted the paraffin and prepared the blocks. Hope this will help you! Ren? J. foley1 wrote: Does anyone routinely allow for the hot wax to drain off multiple cassettes of processed tissue and be held at room temperature for multiple days (6 days) before embedding? What would the consequences be to morphology, possible immunohistochemistry and molecular (DNA) studies? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------- LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Fri Jan 6 12:12:35 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Jan 6 12:12:48 2006 Subject: [Histonet] Processing/embedding question - your opinion In-Reply-To: <6.2.3.4.0.20060106120242.03b2f4d0@ander093.email.umn.edu> Message-ID: <20060106181235.22386.qmail@web61224.mail.yahoo.com> To all that have been offended by my opinion I apologize. It was not my intention of offending anybody, I was just expressing my opinion that remains the same as initially stated. The heading of the initial posting reads:"-your opinion", well that is my opinion! I just think that it takes the same amount of work to prepare a large block with all the cassettes in, than wrapping them, or putting them aside. Ren? J. (by the Ren?e is a female name, Ren? is male!) LuAnn Anderson wrote: Why not do a comparison--take two pieces of the same tissue. Process both-embed one immediately and delay the second. Cut, stain and compare morphology. LuAnn At 11:54 AM 1/6/2006, Trajkovic, Dusko wrote: >I have yet to see any published information >stating that un-embedded tissue left to >solidify, is damaging in any way or form. My >colleagues an I in research, have done it >numerous times, for various reasons, non were for being lazy. > >Dusko Trajkovic > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa >Sent: Friday, January 06, 2006 9:43 AM >To: Bonnie Whitaker; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Processing/embedding question - your opinion > >Bonnie: > That is my personal and very particular > opinion, and I think I am entitled to that, > in the same way that you have expressed yours. > I believe that this is covered by the freedom of expression, is it not? > Ren? J. > >Bonnie Whitaker wrote: > Rene, > >What is "lazy and indolent" about saving processed tissue without embedding >it? I frequently collect many tissues for embedding in multi-tissue blocks, >and I see no point in embedding the tissue temporarily while I continue to >collect additional material. It makes no difference if the paraffin coating >is minimal or a full block. You can also store a lot more control tissue in >a small area if it is maintained in unembedded cassettes. One cassette can >hold several blocks worth of material. > >Bonnie Whitaker >Lab Manager >Brown & Associates Medical Laboratories >8076 El Rio >Houston, Texas 77054 >713-741-6677 > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa >Sent: Friday, January 06, 2006 10:58 AM >To: foley1; histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Processing/embedding question - your opinion > > >Hi Foley: >I have seen that done in several places. I have also seen all the >cassettes wrapped in aluminum foil and kept that way (like a pizza >left-over). >I cannot tell you what would be the consequences for the morphology or >future tests. >Theoretically speaking probably this practice will not affect because the >tissue itself and all its components are supposedly embedded in paraffin >that will just solidify. >I personally do not like this to be done. For me, personally, it indicates >laziness, indolence and even "disrespect" for the tissue sample. >Once I was confronted with the need of keeping processed cassettes in a >secure way before casting the blocks. My solution was to put all the >cassettes in a shallow >plastic container, place all the blocks in it, add melted paraffin and >prepare one single block, as large as the container. When I was able to >prepare the blocks individually, I melted the paraffin and prepared the >blocks. >Hope this will help you! >Ren? J. > >foley1 wrote: >Does anyone routinely allow for the hot wax to drain off multiple >cassettes of processed tissue and be held at room temperature for multiple >days (6 >days) before embedding? What would the consequences be to morphology, >possible immunohistochemistry and molecular (DNA) studies? > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > >--------------------------------- >Yahoo! DSL Something to write home about. Just $16.99/mo. or less >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > >--------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >---------------------------------------------------------------------- >LEGAL NOTICE >Unless expressly stated otherwise, this message >is confidential and may be privileged. It is >intended for the addressee(s) only. Access to >this E-mail by anyone else is unauthorized. If >you are not an addressee, any disclosure or >copying of the contents of this E-mail or any >action taken (or not taken) in reliance on it is >unauthorized and may be unlawful. If you are >not an addressee, please inform the sender immediately. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From jennifer.l.hofecker <@t> Vanderbilt.Edu Fri Jan 6 12:22:09 2006 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Fri Jan 6 12:26:47 2006 Subject: [Histonet] Microwave slide drying vs. conventional slide drying References: <5B3B26C4B71D5E469892D1860ABE10EA7F1E7F@SHSEXVS01.int.samhealth.net> Message-ID: <898D946569A27444B65667A49C074052852B3C@mailbe06.mc.vanderbilt.edu> Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax I have done both but I agree with the person that just used the hot plate, it's very easy and really works in a brief amount of time. There is some science behind why you should not heat paraffin slides in the microwave. The late (and great) Steve Slapp used to talk about it in his workshops, but I can't think of it right now. It was something molecular... Does anybody know what the scientific reason is? And before I get flamed, I am asking because I know there are many microwave gurus out there and not because I am too lazy to look it up. Have a great weekend, Jennifer ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of lharris@samhealth.org Sent: Fri 1/6/2006 10:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave slide drying vs. conventional slide drying We are having a discussion about slide drying at out facility. How many of you out there use a microwave to dry your H&E slides before staining versus using a conventional 70 degree Celsius hot air slide drying oven for 20 minutes? I personally against microwave slide drying but the doctors like it because it takes less time (3 min. 30 sec. on medium setting). Thanks for your response. Lori A. Harris, HT (ASCP) Histology Section Leader GSRMC - Pathology 3600 NW Samaritan Drive Corvallis, OR 97330 1-541-768-6078 lharris@samhealth.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 From BGapinski <@t> pathgroup.com Fri Jan 6 12:36:36 2006 From: BGapinski <@t> pathgroup.com (Bruce Gapinski) Date: Fri Jan 6 12:37:43 2006 Subject: [Histonet] Number of GI biopsy sections on a slide Message-ID: We have a protocol where we automatically give the Pathologists 4 levels (really 3 'cause one is just a "full face"). We do this based on the amount of material submitted and only for specific tissues. G.I. skins, breast core bxs, cervix, prostate needle bxs, any nasopharyngeal bxs and a few others I can't remember off the top of my head. Peace, Bruce Gapinski --------------------------------------------------------------------- Important Notice: This e-mail is intended for the use of the person to whom it is addressed and may contain information that is privileged and confidential. If you are not the intended recipient, any disclosure, copying, distribution, or use of the contents of this message is strictly prohibited. If you have received this e-mail in error, please destroy this message and contact the Security Officer at PathGroup, Inc. immediately at 615-221-4431. From TJJ <@t> Stowers-Institute.org Fri Jan 6 13:10:53 2006 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Jan 6 13:11:23 2006 Subject: [Histonet] Re: 50 micron mouse brain Message-ID: Atoska, Good luck getting anything that isn't a roll. We've successfully done 30 micron sections of samples here. One thing you might try is floating the paraffin block in warm (~40-45 degreeC) water for about 10-15 seconds and then taking your section (slowly). You'll probably only be good for one section before it rolls again. It's worth a shot Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From jkiernan <@t> uwo.ca Fri Jan 6 13:24:50 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Jan 6 13:24:58 2006 Subject: [Histonet] Processing/embedding question - your opinion References: Message-ID: <43BEC402.CE86E8CF@uwo.ca> Which is worst? Being lazy, being indolent or being both at the same time? Oh, to be idle! John Kiernan London, Canada. --------------------- "Allen, Rhonda" wrote: > > Renee, > It may be your opinion, but to those of us who have done it out of necessity, not being "lazy and indolent", it sounds like you are trying to make us feel "lazy and indolent". > > Rhonda Allen BA HT(ASCP)HTL, QIHC > Histotechnology Specialist II > Stowers Institute > 1000 E. 50th Street > Kansas City, MO 64110 > 816-926-4305 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa > Sent: Friday, January 06, 2006 11:43 AM > To: Bonnie Whitaker; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Processing/embedding question - your opinion > > Bonnie: > That is my personal and very particular opinion, and I think I am entitled to that, > in the same way that you have expressed yours. > I believe that this is covered by the freedom of expression, is it not? > Ren? J. > > Bonnie Whitaker wrote: > Rene, > > What is "lazy and indolent" about saving processed tissue without embedding it? I frequently collect many tissues for embedding in multi-tissue blocks, and I see no point in embedding the tissue temporarily while I continue to collect additional material. It makes no difference if the paraffin coating is minimal or a full block. You can also store a lot more control tissue in a small area if it is maintained in unembedded cassettes. One cassette can hold several blocks worth of material. > > Bonnie Whitaker > Lab Manager > Brown & Associates Medical Laboratories > 8076 El Rio > Houston, Texas 77054 > 713-741-6677 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa > Sent: Friday, January 06, 2006 10:58 AM > To: foley1; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Processing/embedding question - your opinion > > Hi Foley: > I have seen that done in several places. I have also seen all the cassettes wrapped in aluminum foil and kept that way (like a pizza left-over). I cannot tell you what would be the consequences for the morphology or future tests. Theoretically speaking probably this practice will not affect because the tissue itself and all its components are supposedly embedded in paraffin that will just solidify. I personally do not like this to be done. For me, personally, it indicates laziness, indolence and even "disrespect" for the tissue sample. Once I was confronted with the need of keeping processed cassettes in a secure way before casting the blocks. My solution was to put all the cassettes in a shallow plastic container, place all the blocks in it, add melted paraffin and prepare one single block, as large as the container. When I was able to prepare the blocks individually, I melted the paraffin and prepared the blocks. Hope this will help you! Ren? J. > > foley1 wrote: > Does anyone routinely allow for the hot wax to drain off multiple cassettes of processed tissue and be held at room temperature for multiple days (6 > days) before embedding? What would the consequences be to morphology, possible immunohistochemistry and molecular (DNA) studies? > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > --------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > --------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GDawson <@t> dynacaremilwaukee.com Fri Jan 6 13:34:47 2006 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Jan 6 13:37:19 2006 Subject: [Histonet] HTL certification Message-ID: Hermina, Two of my techs just took the HTL yesterday & passed. According to them, the written test was a nightmare. Nothing came from the old stand-by textbooks like Sheehan or Carson. The BOR study guide provided zero help. There was alot of IHC related questions that included trouble shooting and appropriate controls for certain stains. One or two ISH questions and 3 or 4 EM questions. They highly recommend studying the anatomy of the eye. Many poor quality pictures of unknown tissues were on it as well along with some strange questions. I believe that their tests were quite different based on how good or bad they were performing. Many managerial questions including worker relations/performance scenarios, OSHA regulations, JHACO rules & CAP regulation. Most of the test dealt with topics that they were not familiar with as they are students with little bench experience. I'm thinking that the final scoring must be on a radical curve because both were sure that they had failed by the end of it. I know for a fact that they couldn't have studied harder than they did for the test, but nothing that they actually studied was on it. Sorry for the gloom and doom, I'm just passing on the sentiments of 2 individuals that just took the test. Good Luck, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI -----Original Message----- From: Hermina Borgerink [mailto:hborgeri@wfubmc.edu] Sent: Friday, January 06, 2006 11:23 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HTL certification Two of my techs will be taking the HTL certification this year. I checked the requirements on the ASCP Board of Registry home page and found that they needed experience in the following areas: fixation, processing, microtomy, and routine as well as special staining. I just wanted to double check with those of you who have recently taken their Board Certifications that EM and ISH experience are no longer required. When I did my HTL certification there still were questions pertaining to both topics, and I just want to make sure they won't be faced with questions they are not trained in. Any response would be greatly appreciated and I thank you in advance. Hermina Hermina M. Borgerink, BA, HT(ASCP)HTL, QIHC Wake Forest University Health Sciences Department of Pathology Medical Center Blvd. Winston-Salem, NC 27157 Tel. (336) 716-1538 Fax. (336) 716-1515 e-mail hborgeri@wfubmc.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Fri Jan 6 13:38:52 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Jan 6 13:39:21 2006 Subject: [Histonet] Processing/embedding question - your opinion Message-ID: Hello Histonetters, Nobody is calling anyone lazy, so just let it go and have a nice day? :>) Thanks Robyn OHSU From jwatson <@t> gnf.org Fri Jan 6 13:45:22 2006 From: jwatson <@t> gnf.org (James Watson) Date: Fri Jan 6 13:45:37 2006 Subject: [Histonet] 50 micron mouse brain Message-ID: Atoska, I our lab we are able to cut 250 micron paraffin sections of Golgi stained whole mouse brains. We add 10% glycerin in our absolute alcohols on the processor for the Golgi stain to keep the tissue soft (we use 5% glycerin in our absolute alcohols on all our animal tissue processing- this shortens our soaking times greatly, eliminates the need to chill blocks, and eliminates cracking or drying artifact in the animal tissue). Then we section the blocks on a sliding microtome, with a long soak of warm water, moving the knife through the block very slowly. We also cut 20-50 micron sections on our rotary microtomes of mouse brains that have been processed with the 5% glycerin, but that does require long soaking times. Good luck. James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Atoska S. Gentry Sent: Thursday, January 05, 2006 2:36 PM To: Histonet Subject: [Histonet] 50 micron mouse brain Hello, will someone please give me pointers on obtaining good 50u sections of PFA drop fixed and/or immersion fixed coronal paraffin sections of mouse brain? As I section they tend to roll up in tight rolls. I've tried sectioning at room temp. vs. chilled sections. Any assistance provided will be greatly appreciated. Thanks, Atoska _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From david.kinsley <@t> spcorp.com Fri Jan 6 13:57:15 2006 From: david.kinsley <@t> spcorp.com (Kinsley, David) Date: Fri Jan 6 13:57:38 2006 Subject: [Histonet] Microwave slide drying vs. conventional slide dryi ng Message-ID: Can you let me know what tissues samples and temperature you typically use with the hot plate method? I work with animal tissues and usually place my slides on a hot plate at about 45-50 degrees to flatten the sections out (about 1 hour) and then follow by heating them in a 60 degree oven for 4 hours to overnight depending on the sample. Dave -----Original Message----- From: Laurie Colbert [mailto:laurie.colbert@huntingtonhospital.com] Sent: Friday, January 06, 2006 12:38 PM To: lharris@samhealth.org; Histonet (E-mail) Subject: RE: [Histonet] Microwave slide drying vs. conventional slide drying Lori, We don't use either. We simply put our drained slides on the hot plate for about 5 minutes. In previous jobs, we always used an oven, and when I came here I was really nervous about just using the hot plate - but it works! Laurie Colbert Huntington Hospital Pasadena, CA -----Original Message----- From: lharris@samhealth.org [mailto:lharris@samhealth.org] Sent: Friday, January 06, 2006 8:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave slide drying vs. conventional slide drying We are having a discussion about slide drying at out facility. How many of you out there use a microwave to dry your H&E slides before staining versus using a conventional 70 degree Celsius hot air slide drying oven for 20 minutes? I personally against microwave slide drying but the doctors like it because it takes less time (3 min. 30 sec. on medium setting). Thanks for your response. Lori A. Harris, HT (ASCP) Histology Section Leader GSRMC - Pathology 3600 NW Samaritan Drive Corvallis, OR 97330 1-541-768-6078 lharris@samhealth.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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From GDawson <@t> dynacaremilwaukee.com Fri Jan 6 14:13:12 2006 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Jan 6 14:15:44 2006 Subject: [Histonet] Processing/embedding question - your opinion Message-ID: I don't know about anyone else, but when I want to disrespect my tissue samples, I say terrible things about their mothers. Har har, Glen Dawson -----Original Message----- From: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: Friday, January 06, 2006 11:43 AM To: Bonnie Whitaker; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Processing/embedding question - your opinion Bonnie: That is my personal and very particular opinion, and I think I am entitled to that, in the same way that you have expressed yours. I believe that this is covered by the freedom of expression, is it not? Ren? J. Bonnie Whitaker wrote: Rene, What is "lazy and indolent" about saving processed tissue without embedding it? I frequently collect many tissues for embedding in multi-tissue blocks, and I see no point in embedding the tissue temporarily while I continue to collect additional material. It makes no difference if the paraffin coating is minimal or a full block. You can also store a lot more control tissue in a small area if it is maintained in unembedded cassettes. One cassette can hold several blocks worth of material. Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, January 06, 2006 10:58 AM To: foley1; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Processing/embedding question - your opinion Hi Foley: I have seen that done in several places. I have also seen all the cassettes wrapped in aluminum foil and kept that way (like a pizza left-over). I cannot tell you what would be the consequences for the morphology or future tests. Theoretically speaking probably this practice will not affect because the tissue itself and all its components are supposedly embedded in paraffin that will just solidify. I personally do not like this to be done. For me, personally, it indicates laziness, indolence and even "disrespect" for the tissue sample. Once I was confronted with the need of keeping processed cassettes in a secure way before casting the blocks. My solution was to put all the cassettes in a shallow plastic container, place all the blocks in it, add melted paraffin and prepare one single block, as large as the container. When I was able to prepare the blocks individually, I melted the paraffin and prepared the blocks. Hope this will help you! Ren? J. foley1 wrote: Does anyone routinely allow for the hot wax to drain off multiple cassettes of processed tissue and be held at room temperature for multiple days (6 days) before embedding? What would the consequences be to morphology, possible immunohistochemistry and molecular (DNA) studies? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Fri Jan 6 16:32:36 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Jan 6 16:32:45 2006 Subject: [Histonet] Microwave slide drying vs. conventional slide drying In-Reply-To: <5B3B26C4B71D5E469892D1860ABE10EA7F1E7F@SHSEXVS01.int.samhealth.net> Message-ID: <20060106223236.65087.qmail@web61218.mail.yahoo.com> Hi Lori: When you place slides with sections in the microwaves oven (MWO) you are not heating the paraffin, you are heating the slides. Paraffins are non polar molecules and do not vibrate under the microwaves (MW), they are "blind" to the paraffin that has a penetration ratio of 15,000 cm. MW go through the paraffin as if it did not exist at all. That is not the case of water that has a penetration ratio of just 4.4 cm. This means that placing the slides in a MWO you will melt the paraffin but because the glass in the slides heats (and could break because glass, as you know, if a very bad heat conductor). For this reason some tissue processors that use MW (like the "newest" Sakura) has 3 chambers, the first 2 heated under the influence of MW and the last, where the paraffin rests, is heated electrically; the infiltration is due to the effect of pressure and vacuum. You should heat your slides either in a hot plate (has to be really big to accomodate a large number of slides) or in a conventional convection oven (I prefer this method). The Sakura stainer heats the slides at 70?C during just 7 minutes in a heating chamber before taking the slides to the first xylene. I hope this helps you to argue your case with your doctors. Ren? J. lharris@samhealth.org wrote: We are having a discussion about slide drying at out facility. How many of you out there use a microwave to dry your H&E slides before staining versus using a conventional 70 degree Celsius hot air slide drying oven for 20 minutes? I personally against microwave slide drying but the doctors like it because it takes less time (3 min. 30 sec. on medium setting). Thanks for your response. Lori A. Harris, HT (ASCP) Histology Section Leader GSRMC - Pathology 3600 NW Samaritan Drive Corvallis, OR 97330 1-541-768-6078 lharris@samhealth.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 --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less From alasdair.wilson <@t> blueyonder.co.uk Fri Jan 6 17:26:21 2006 From: alasdair.wilson <@t> blueyonder.co.uk (alasdair wilson) Date: Fri Jan 6 17:26:33 2006 Subject: [Histonet] RE Processing/Embedding References: Message-ID: <001301c61318$9a9f93d0$114d2952@yourc837daffeb> We often take cassettes off the processor and leave them to solidify while other duties are carried out. We do this out of necessity when busy and then melt the wax and embed the tissues at a convenient time. We have never had any problems with morphology or shrinkage. Alasdair Wilson ----- Original Message ----- From: To: Sent: Friday, January 06, 2006 5:58 PM Subject: Histonet Digest, Vol 26, Issue 4 > 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: document for recording special stain results for quality > control (Rene J Buesa) > 2. Smear preps (Loralei Dewe) > 3. Re: document for recording special stain results for quality > control (Linda Blazek) > 4. Re:Plastic sectioning (Gayle Callis) > 5. RE: Happy New Year's and competancy/volume questions > (Kinsley, David) > 6. gross room (Tom Wells) > 7. 50 micron mouse brain (Atoska S. Gentry) > 8. Job Opportunity (lena spencer) > 9. Re: Number of GI biopsy sections on a slide for > pediatrichospitals. (Joe Nocito) > 10. Re: CD4 and CD8 in mouse / human FFPE (Kuo Christy) > 11. RE: (Louis Apuzzio) > 12. RE: optimal thickness for cutting of IHC sections > (C.M. van der Loos) > 13. Looking for a home... (Ford Royer) > 14. slide dryer (dfleisch@bidneedham.org) > 15. Processing/embedding question - your opinion (foley1) > 16. RE: Processing/embedding question - your opinion (Allen, Rhonda) > 17. RE: Price quotes on refurbished lab equipment/ Start Up Lab > (mprice26@juno.com) > 18. Re: Processing/embedding question - your opinion (Rene J Buesa) > 19. Microwave slide drying vs. conventional slide drying > (lharris@samhealth.org) > 20. sucrose liver (Steven Coakley) > 21. HTL certification (Hermina Borgerink) > 22. RE: Processing/embedding question - your opinion (Bonnie Whitaker) > 23. RE: Microwave slide drying vs. conventional slide drying > (Laurie Colbert) > 24. RE: Processing/embedding question - your opinion (Rene J Buesa) > 25. RE: Processing/embedding question - your opinion (Allen, Rhonda) > 26. microwave tissue processing (Harrison, Sandra C.) > 27. RE: Processing/embedding question - your opinion > (Trajkovic, Dusko) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Thu, 5 Jan 2006 10:00:23 -0800 (PST) > From: Rene J Buesa > Subject: Re: [Histonet] document for recording special stain results > for quality control > To: "Adams, Nancy" , > histonet@lists.utsouthwestern.edu > Message-ID: <20060105180023.17687.qmail@web61221.mail.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > Hi Nancy: > We used to "batch" special stains (1 positive control for slides with the > same procedure, either IHC, HC or even Her-2new), so we had a "common" > form where we used to write the date, the test we batched, and the cases > numbers included in the batch. > We checked if the control reacted as expected, attached the form and the > control to the batched slides and gave them to the pathologist. > The form was originally signed by the histotech who did the test and it > was also signed by the pathologist. > All the forms were filed by the type of test (Steiner, IHCs on the day, > PAS, etc.) and were available for inspection. > In our last CAP inspection we were told that this was a good procedure. > I hope this will help you! > Ren? J. > > "Adams, Nancy" wrote: > Does anyone have a good document for recording this information that > they'd be willing to share? I'm in the process of getting ready for a > CAP inspection. > > Thanks > > Nancy Rutledge > > Falmouth Hospital > > Falmouth, > > MA > > > > ************************************************************ > 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 contact the > system administrator for Cape Cod Healthcare. > Administrator@CapeCodHealth.org > ************************************************************ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > --------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less > > ------------------------------ > > Message: 2 > Date: Thu, 5 Jan 2006 10:11:58 -0800 (PST) > From: Loralei Dewe > Subject: [Histonet] Smear preps > To: "Van Eyck, Deb" > Cc: "' histonet@lists.utsouthwestern.edu'" > > Message-ID: > Content-Type: TEXT/PLAIN; charset=US-ASCII > > > I need to do some smears from frozen tissue. I've never done them before > but they don't sound all that tough. Anyone got a good protocol to share > with me? > > Cheers, > Loralei Dewe > UC Davis Vet Hospital > > > > ------------------------------ > > Message: 3 > Date: Thu, 05 Jan 2006 13:12:52 -0500 > From: "Linda Blazek" > Subject: Re: [Histonet] document for recording special stain results > for quality control > To: nadams@CapeCodHealth.org, histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset=US-ASCII > > I forgot to add something to my previous note. We run batch controls also > as Rene does but we have a control label ie: GMS #1, IRON #1 etc. The > control number is also dictated on the pathology report. That way we can > always go back to the correct control slide - Linda > > > Linda Blazek, HT (ASCP) > Department of Pathology > Children's Medical Center > Dayton, Ohio 45404 > (937) 641-3358 > fax (937)641-5482 > blazekl@childrensdayton.org > > >>>> "Adams, Nancy" 01/05/2006 11:13 AM >>> > > Does anyone have a good document for recording this information that > they'd be willing to share? I'm in the process of getting ready for a > CAP inspection. > > Thanks > > Nancy Rutledge > > Falmouth Hospital > > Falmouth, > > MA > > > > ************************************************************ > 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 contact the > system administrator for Cape Cod Healthcare. > Administrator@CapeCodHealth.org > ************************************************************ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 4 > Date: Thu, 05 Jan 2006 13:22:24 -0700 > From: Gayle Callis > Subject: [Histonet] Re:Plastic sectioning > To: , Histonet@lists.utsouthwestern.edu > Message-ID: > <6.0.0.22.1.20060105130946.01b60008@gemini.msu.montana.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Sunny, > > You have a tough problem here. 30 um thick GMA sections may be too thick > for this embedding media. GMA plastic was initially designed for doing > THIN sections at 1 to 3 micrometers. It tends to be brittle when cutting > thick sections and one might have to use a very sharp tungsten carbide > tipped knife in order to get any thick section off a GMA block. One could > try to adjust the plasticizer by increasing its concentration to make the > GMA more pliable. The knife, no matter how sharp, may not actually slice > through the plastic, but force its way through in such a way that the > plastic shatters i.e. a shearing effect and not really have a clean cut. > > Patsy Ruegg published an inhouse method for GMA, rather than buy kits - > that way she could adjust the ingredients accordingly for softer or harder > GMA. > > If the tissues are thicker than 1 to 2 mm when processing and embedding, > infiltration may be a problem unless extended time is used and done at 4C > ( > refrigerator) and polymerization might have to controlled with cold > temperatures in order to slow it down for continuous or steady progression > of polymerizing. That can be done at 4C also. > > At 12:58 AM 1/5/2006, you wrote: >>Dear Sunny, >> >>First about the knife marks. To me it seems that you are using a blunt >>knife. Try a new/sharp one. >>About the shattering: I don't know how youre blocks look like. Is the GMA >>well impregnated into the brain-tissue, for else this could explain the >>problem. If this is not the case (I don't think, else you would have >>mentioned it) then perhaps you can try to use some sort of tape. Stick it >>onto the block and then cut you're slice. I've tried it and it worked in >>my case. The only problem then is how to remove the tape without damaging >>the slice (unless you stain the slices when still sticked to the tape). >>There are also commercial tapes specially for this technique (see for >>instance this link: >>http://www.alphelys.com/site/us/pHGP_CoupesParaffine.htm). > > Gayle Callis > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 > 406 994-4303 (FAX) > > > > > > ------------------------------ > > Message: 5 > Date: Thu, 5 Jan 2006 16:32:50 -0500 > From: "Kinsley, David" > Subject: RE: [Histonet] Happy New Year's and competancy/volume > questions > To: 'Rene J Buesa' , "Van Eyck, Deb" > , "' histonet@lists.utsouthwestern.edu'" > > Message-ID: > > Content-Type: text/plain; charset="iso-8859-1" > > Hi all, > > It seems to me that the competencies below are for human tissues in a > hospital/clinical setting. I was wondering if there are > competency/productivity standards for research histology on animal tissues > particularly mouse and rat species, paraffin and frozen. We are looking > to > establish some base guidelines for our lab. Any suggestions are > appreciated. > > thanks > > Dave > > > -----Original Message----- > From: Rene J Buesa [mailto:rjbuesa@yahoo.com] > Sent: Wednesday, January 04, 2006 6:33 PM > To: Van Eyck, Deb; ' histonet@lists.utsouthwestern.edu' > Subject: Re: [Histonet] Happy New Year's and competancy/volume questions > > > Hi Deb: > In 3 labs I wrote competencies for the expectation for sectoning was set > at 25 blocks/hour and for embedding at 60 blocks/hour. > Each histotech was evaluated on that basis and their individual averages > were > rated (as percentages) against the expectation. Those with averages above > expectations were evaluated better (and received salary compensations > accordingly). Nobody was considered "proficient" until their individual > average reached the expectation consistently. > For time utilization (ratio between time with recorded tasks versus > logged > time) the expectation was 75 % (i.e. 6 h with logged tasks agains 8 h/day > per shift). > In the Dec./04 issue of the Journal of Histotechnology there are averages > for every imaginable task within the histology lab as a result of a survey > of 12 labs. with annual volumes ranging from 4800 to 77000 cases/year. If > your lab is within that range this information could be used by you. > I hope this will help you! > Ren? J. > > "Van Eyck, Deb" wrote: > > Hi all, > > If anyone has information they would like to share I would appreciate it. > We > are a hospital lab and we are updating doing competencies. We are > interested > in Volumes and expectations for embedding, cutting etc, that take into > account differeing experience levels, training and quality > expectations---does anyone have a good system or have good references or > articles that discuss this issue? thanks for your input. Deb > > > This information is confidential and intended solely for the use of the > individual or entity to whom it is addressed. > If you have received this email in error please notify the sender or our > Customer Support Center at (262) 928-2777. > > We have scanned this email and its attachments for malicious content. > However, the recipient should check this email and any attachments for the > presence of viruses. ProHealth Care accepts no liability for any damage > caused by any virus transmitted by this email. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > --------------------------------- > Yahoo! Photos > Ring in the New Year with Photo Calendars. Add photos, events, holidays, > whatever. _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > ********************************************************************* > This message and any attachments are solely for the intended recipient. If > you are not the intended recipient, disclosure, copying, use or > distribution of the information included in this message is prohibited -- > Please immediately and permanently delete. > > > > > ------------------------------ > > Message: 6 > Date: Thu, 5 Jan 2006 14:06:00 -0800 > From: Tom Wells > Subject: [Histonet] gross room > To: histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset="US-ASCII" > > I am going to be discussing the gross room and prosecting with my medical > laboratory technology students soon. This is an area that seems to be > rather poorly documented. Do any of you have gross room manuals that you > wouldn't mind sharing with me? Are there any web sites which discuss > grossing for new residents? Are there any pathology sites that have > established standards for grossing etc. Any help would be gratefully > appreciated. Thanks. Tom > > ------------------------------ > > Message: 7 > Date: Thu, 05 Jan 2006 16:36:05 -0600 > From: "Atoska S. Gentry" > Subject: [Histonet] 50 micron mouse brain > To: Histonet > Message-ID: > <6.0.1.1.0.20060105163140.01acf798@mailhost.vetmed.auburn.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Hello, will someone please give me pointers on obtaining good 50u sections > of PFA drop fixed and/or immersion fixed coronal paraffin sections of > mouse > brain? As I section they tend to roll up in tight rolls. I've tried > sectioning at room temp. vs. chilled sections. Any assistance provided > will > be greatly appreciated. Thanks, Atoska > > > > > ------------------------------ > > Message: 8 > Date: Thu, 5 Jan 2006 18:55:31 -0500 > From: "lena spencer" > Subject: [Histonet] Job Opportunity > To: > Message-ID: <005401c61253$8311b880$3379df0c@org.insightbb.com> > Content-Type: text/plain; charset="iso-8859-1" > > Electron Microscopy Job Opportunity > Full Time/Day Shift > some histology duties required > > Norton Healthcare > Louisville, Kentucky > Contact: Mary Beth Knight, MT(ASCP)HTL > marybeth.knight@nortonhealthcare.org > 502-629-7884 > > > > > ------------------------------ > > Message: 9 > Date: Thu, 5 Jan 2006 21:35:07 -0600 > From: "Joe Nocito" > Subject: Re: [Histonet] Number of GI biopsy sections on a slide for > pediatrichospitals. > To: "Foshey, Annette" , > > Message-ID: <007901c61272$30e4f1c0$e8bd0b43@yourxhtr8hvc4p> > Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=original > > Annette, > we cut 3 levels on one slide with 2 sections per level > > Joe Nocito BS, HT(ASCP)QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > ----- Original Message ----- > From: "Foshey, Annette" > To: > Sent: Wednesday, January 04, 2006 3:50 PM > Subject: [Histonet] Number of GI biopsy sections on a slide for > pediatrichospitals. > > > > I am inquiring what the "standard" is for the number of GI biopsy > sections that are routinely put on slides for review. We currently cut > two slides with two levels on each slide. The total number of sections > is between 6-8 sections on each slide. > > Thanks, > Annette Foshey, HT > Team Leader in Histology > Children's Hospital of Wisconsin > > > ************************** > Children's Hospital and Health System recognizes that unencrypted e-mail > is > insecure 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 > > > -- > No virus found in this incoming message. > Checked by AVG Free Edition. > Version: 7.1.371 / Virus Database: 267.14.13/221 - Release Date: 1/4/2006 > > > > > > ------------------------------ > > Message: 10 > Date: Fri, 6 Jan 2006 14:35:02 +0800 > From: Kuo Christy > Subject: [Histonet] Re: CD4 and CD8 in mouse / human FFPE > To: Laurie Reilly > Cc: histonet , > wasielewski.reinhard.von@mh-hannover.de > Message-ID: > <5e4834130601052235w191fb3f9g206edcd8ac567a9e@mail.gmail.com> > Content-Type: text/plain; charset=ISO-8859-1 > > Hi Reinhard, > Could I also ask for a copy of your protocol, cuz the results from > anti-CD4(Novocvastra) seems to be very unspecific on human tissue > FFPE sections and I don't know the reason. > > Thank you for your help! > > Best wishes, > Christy > christy0424@gmail.com > > On 12/19/05, Laurie Reilly wrote: >> Reinhard, >> Could I please have a copy of your protocol, we have had lots of trouble >> trying to stain with >> CD4 and CD8 in bovine tissues. >> Any advice you can give would be most welcome, particularly about antigen >> retrieval. >> >> Hope everyone has a happy Christmas, >> Regards, >> Laurie. >> >> At 08:49 PM 18/12/2005 +0100, you wrote: >> >Hi Patsy, >> >we do a lot of CD4 and CD8 in human FFPE tissues, even in BM after >> >decalification. >> >It works very good and is daily routine in our lab. If you need advice, >> >give me a mail. >> >We now got some hints how both markers could work in mouse tissue, too. >> >If >> we >> >succeed, we'll let you know. >> > >> >best regards >> >Reinhard. >> > >> > >> > >> >PD Dr. med. Reinhard von Wasielewski >> > >> > >> >_______________________________________________ >> >Histonet mailing list >> >Histonet@lists.utsouthwestern.edu >> >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> Mr.Laurie Reilly Ph 07 4781 >> 4468 >> School of Veterinary & Biomedical Science Fax 07 4779 1526 >> James Cook University >> Townsville Qld. >> 4811 laurie.reilly@jcu.edu.au >> >> Australia. >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > ------------------------------ > > Message: 11 > Date: Fri, 6 Jan 2006 05:54:04 -0500 > From: "Louis Apuzzio" > Subject: [Histonet] RE: > To: > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > Anyone in search of a Histotech, ( HT eligible) and also a Cytotech, ( > CT(ASCP), > > for small private laboratory and or hospital, please contact: > > ljapuzzio@msn.com > > Thank you ! > > > ------------------------------ > > Message: 12 > Date: Fri, 06 Jan 2006 12:26:41 +0100 > From: "C.M. van der Loos" > Subject: [Histonet] RE: optimal thickness for cutting of IHC sections > To: histonet@lists.utsouthwestern.edu > Cc: PMonfils@Lifespan.org > Message-ID: <1d9a431dc72b.1dc72b1d9a43@amc.uva.nl> > Content-Type: text/plain; charset="windows-1252" > > > Sorry guys my message yesterday was somewhat distorted. Here it is > again... > > > Dear Paul, > > > I totally agree what you wrote about IHC: it's just a surface event. > > Long time ago we had to prove that our (prehistoric) attempt of > quantify due to microtome different thickness and gue on the > staining intensity. The only thicker sections showed a higher > non-specific b than thinner sections. See: > > > CM van der Loos, MMH Marijianowski and AE Becker: Quantification in > immu collagen types I > > Chris van der Loos, PhD > Dept. of Pathology > Academic Medical Center Meibergdreef 9 > NL-1105 AZ Amsterdam > The Netherlands > > > > > > Date: Tue, 3 Jan 2006 13:09:20 -0500 > From: "Monfils, Paul" thickness for cutti sections > To: [1]'histonet@lists.utsouthwestern.edu' I believe section > thickness is less critical for IHC because antibo dies are > very large molecules that don't penetrate tissue very well, so > regardless of > the thickness of the section, you are really only staining > exposed > surface of the tissue, perhaps to a depth of 2 microns or s have > verified this by electron microscopy. This is qui standard > histochemical procedures where a thicker sect intense > stain because the small dye molecules pene and stain > it all the way through. > > > > > > > > > // Script broken into two blocks. There is a in the > document.write bugs 4553030/31, 5047193. document.form > var form1 = document.form1 > var form2 = > document.form2 > var errno = 0 > arg=window.location.search;//AK > if (!refreshing) { > if > (arg.length > 6 && arg.substring(0, 7) == 'bgcolor') > color(arg.substring(8, arg.length)) > f = eval('parent.' + > self.name + 'Init') > if (f) > f() > } > // Note: the tag is > defined in main.js and written out when a given is created. > > > /**Enable backspace for text editing*/ > if > (NN>=5){ > setBkspEvtHndlr("input");//specifically for text > elements. checkboxes/r setBkspEvtHndlr("textarea"); > setBkspEvtHndlr("browse"); > } > > > > References > > 1. 3D"javascript:main.compose('new','t='hi > > ------------------------------ > > Message: 13 > Date: Fri, 6 Jan 2006 08:26:31 -0600 > From: "Ford Royer" > Subject: [Histonet] Looking for a home... > To: "Histonet" > Message-ID: <002001c612cd$30f0a230$3900a8c0@fords> > Content-Type: text/plain; charset="us-ascii" > > A friend has asked me to try and find a home for her retired TissueTek VIP > (K-series) Tissue Processors. > These would make a great back-up unit or a basic unit in a start-up lab. > > Contact me Off-List for details. > > ~ Ford > > Ford M. Royer, MT(ASCP) > Minnesota Medical Specialists, Inc. > Golden Valley, MN 55427-3601 > 763-542-8725 phone > 888-790-9686 toll free > 763-546-4830 fax > email > > > > ------------------------------ > > Message: 14 > Date: Fri, 6 Jan 2006 09:46:22 -0500 > From: > Subject: [Histonet] slide dryer > To: > Message-ID: > > Content-Type: text/plain; charset="us-ascii" > > I am looking for a small slide dryer. > > Would greatly appreciate any suggestions. > > > > Deborah S. Fleischhacker, M.D. > > Pathologist > > Beth Israel Deaconess Hospital Needham > > 148 Chestnut Street > > Needham, MA 02492 > > Phone: (781) 453-3087 > > Fax: (781) 453-3097 > > > > > > ------------------------------ > > Message: 15 > Date: Thu, 29 Dec 2005 12:40:59 -0500 > From: foley1 > Subject: [Histonet] Processing/embedding question - your opinion > To: > Message-ID: > Content-Type: text/plain; charset="US-ASCII" > > Does anyone routinely allow for the hot wax to drain off multiple > cassettes > of processed tissue and be held at room temperature for multiple days (6 > days) before embedding? What would the consequences be to morphology, > possible immunohistochemistry and molecular (DNA) studies? > > > > > > ------------------------------ > > Message: 16 > Date: Fri, 6 Jan 2006 10:19:39 -0600 > From: "Allen, Rhonda" > Subject: RE: [Histonet] Processing/embedding question - your opinion > To: "foley1" , > > Message-ID: > > > Content-Type: text/plain; charset="us-ascii" > > Have done it on many occasions and never had it be a problem. > > Rhonda Allen > Stowers Institute > 1000 E. 50th street > Kansas City, Missouri 64110 > rra@stowers-institute.org > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of foley1 > Sent: Thursday, December 29, 2005 11:41 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Processing/embedding question - your opinion > > > Does anyone routinely allow for the hot wax to drain off multiple > cassettes of processed tissue and be held at room temperature for > multiple days (6 > days) before embedding? What would the consequences be to morphology, > possible immunohistochemistry and molecular (DNA) studies? > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 17 > Date: Fri, 6 Jan 2006 16:49:27 GMT > From: "mprice26@juno.com" > Subject: [Histonet] RE: Price quotes on refurbished lab equipment/ > Start Up Lab > To: histonet@lists.utsouthwestern.edu > Message-ID: <20060106.085006.9834.259217@webmail48.nyc.untd.com> > Content-Type: text/plain > > Histonetters, > First of all, Happy New Year to everyone in histoland. > > I am looking for companies that can sell all the equipment (refurbished) > needed for a start up lab. I would like a package deal, everything > included, IE Processor, embedding center, microtome, coverslipper, & > waterbath all together. > > Vendors are welcome to respond. > Thank you. > > Marsha Price > > > > > ------------------------------ > > Message: 18 > Date: Fri, 6 Jan 2006 08:57:38 -0800 (PST) > From: Rene J Buesa > Subject: Re: [Histonet] Processing/embedding question - your opinion > To: foley1 , histonet@lists.utsouthwestern.edu > Message-ID: <20060106165738.73387.qmail@web61219.mail.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > Hi Foley: > I have seen that done in several places. I have also seen all the > cassettes wrapped in aluminum foil and kept that way (like a pizza > left-over). > I cannot tell you what would be the consequences for the morphology or > future tests. > Theoretically speaking probably this practice will not affect because the > tissue itself and all its components are supposedly embedded in paraffin > that will just solidify. > I personally do not like this to be done. For me, personally, it > indicates laziness, indolence and even "disrespect" for the tissue sample. > Once I was confronted with the need of keeping processed cassettes in a > secure way before casting the blocks. My solution was to put all the > cassettes in a shallow > plastic container, place all the blocks in it, add melted paraffin and > prepare one single block, as large as the container. When I was able to > prepare the blocks individually, I melted the paraffin and prepared the > blocks. > Hope this will help you! > Ren? J. > > foley1 wrote: > Does anyone routinely allow for the hot wax to drain off multiple > cassettes > of processed tissue and be held at room temperature for multiple days (6 > days) before embedding? What would the consequences be to morphology, > possible immunohistochemistry and molecular (DNA) studies? > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > --------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less > > ------------------------------ > > Message: 19 > Date: Fri, 6 Jan 2006 08:57:49 -0800 > From: > Subject: [Histonet] Microwave slide drying vs. conventional slide > drying > To: > Message-ID: > <5B3B26C4B71D5E469892D1860ABE10EA7F1E7F@SHSEXVS01.int.samhealth.net> > Content-Type: text/plain; charset="iso-8859-1" > > We are having a discussion about slide drying at out facility. How many of > you out there use a microwave to dry your H&E slides before staining > versus using a conventional 70 degree Celsius hot air slide drying oven > for 20 minutes? I personally against microwave slide drying but the > doctors like it because it takes less time (3 min. 30 sec. on medium > setting). Thanks for your response. > > Lori A. Harris, HT (ASCP) > Histology Section Leader > GSRMC - Pathology > 3600 NW Samaritan Drive > Corvallis, OR 97330 > 1-541-768-6078 > lharris@samhealth.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. > > > ------------------------------ > > Message: 20 > Date: Fri, 6 Jan 2006 09:23:14 -0800 (PST) > From: Steven Coakley > Subject: [Histonet] sucrose liver > To: Histonet@lists.utsouthwestern.edu > Message-ID: <20060106172314.29262.qmail@web90204.mail.scd.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > > Good morning everyone, If anyone out there cryo-sections sucrose > infiltrated fixed liver I'd like to know what temp there are sectioning at > best. The mouse liver is fixed at 4C in formalin/PBS for 6 hours the > 25-30% sucrose overnight also at 4C. I thought sucrose infiltrated tissue > should section closer to -25C. Our chatter at anything above -18C. > > Steve > > > --------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less > > ------------------------------ > > Message: 21 > Date: Fri, 6 Jan 2006 12:23:26 -0500 > From: "Hermina Borgerink" > Subject: [Histonet] HTL certification > To: > Message-ID: > <9AEEF1FB6254224AA355ED285F84916515A37AA7@EXCHVS2.medctr.ad.wfubmc.edu> > > Content-Type: text/plain; charset="us-ascii" > > Two of my techs will be taking the HTL certification this year. I > checked the requirements on the ASCP Board of Registry home page and > found that they needed experience in the following areas: fixation, > processing, microtomy, and routine as well as special staining. I just > wanted to double check with those of you who have recently taken their > Board Certifications that EM and ISH experience are no longer required. > When I did my HTL certification there still were questions pertaining to > both topics, and I just want to make sure they won't be faced with > questions they are not trained in. Any response would be greatly > appreciated and I thank you in advance. > Hermina > > > Hermina M. Borgerink, BA, HT(ASCP)HTL, QIHC > Wake Forest University Health Sciences > Department of Pathology > Medical Center Blvd. > Winston-Salem, NC 27157 > Tel. (336) 716-1538 > Fax. (336) 716-1515 > e-mail hborgeri@wfubmc.edu > > > > > > > > > ------------------------------ > > Message: 22 > Date: Fri, 6 Jan 2006 11:32:30 -0600 > From: "Bonnie Whitaker" > Subject: RE: [Histonet] Processing/embedding question - your opinion > To: "'Rene J Buesa'" , > > Message-ID: <000001c612e7$2be04060$3601a8c0@brownpathology.net> > Content-Type: text/plain; charset="iso-8859-1" > > Rene, > > What is "lazy and indolent" about saving processed tissue without > embedding > it? I frequently collect many tissues for embedding in multi-tissue > blocks, > and I see no point in embedding the tissue temporarily while I continue to > collect additional material. It makes no difference if the paraffin > coating > is minimal or a full block. You can also store a lot more control tissue > in > a small area if it is maintained in unembedded cassettes. One cassette > can > hold several blocks worth of material. > > Bonnie Whitaker > Lab Manager > Brown & Associates Medical Laboratories > 8076 El Rio > Houston, Texas 77054 > 713-741-6677 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J > Buesa > Sent: Friday, January 06, 2006 10:58 AM > To: foley1; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Processing/embedding question - your opinion > > > Hi Foley: > I have seen that done in several places. I have also seen all the > cassettes wrapped in aluminum foil and kept that way (like a pizza > left-over). > I cannot tell you what would be the consequences for the morphology or > future tests. > Theoretically speaking probably this practice will not affect because the > tissue itself and all its components are supposedly embedded in paraffin > that will just solidify. > I personally do not like this to be done. For me, personally, it > indicates > laziness, indolence and even "disrespect" for the tissue sample. > Once I was confronted with the need of keeping processed cassettes in a > secure way before casting the blocks. My solution was to put all the > cassettes in a shallow > plastic container, place all the blocks in it, add melted paraffin and > prepare one single block, as large as the container. When I was able to > prepare the blocks individually, I melted the paraffin and prepared the > blocks. > Hope this will help you! > Ren? J. > > foley1 wrote: > Does anyone routinely allow for the hot wax to drain off multiple > cassettes of processed tissue and be held at room temperature for multiple > days (6 > days) before embedding? What would the consequences be to morphology, > possible immunohistochemistry and molecular (DNA) studies? > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > --------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 23 > Date: Fri, 6 Jan 2006 09:38:07 -0800 > From: "Laurie Colbert" > Subject: RE: [Histonet] Microwave slide drying vs. conventional slide > drying > To: , "Histonet (E-mail)" > > Message-ID: > <0BE6ADFAE4E7E04496BF21ABD346628005653929@EXCHANGE1.huntingtonhospital.com> > > Content-Type: text/plain; charset="iso-8859-1" > > Lori, > > We don't use either. We simply put our drained slides on the hot plate > for about 5 minutes. In previous jobs, we always used an oven, and when I > came here I was really nervous about just using the hot plate - but it > works! > > Laurie Colbert > Huntington Hospital > Pasadena, CA > > -----Original Message----- > From: lharris@samhealth.org [mailto:lharris@samhealth.org] > Sent: Friday, January 06, 2006 8:58 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Microwave slide drying vs. conventional slide drying > > > We are having a discussion about slide drying at out facility. How many of > you out there use a microwave to dry your H&E slides before staining > versus using a conventional 70 degree Celsius hot air slide drying oven > for 20 minutes? I personally against microwave slide drying but the > doctors like it because it takes less time (3 min. 30 sec. on medium > setting). Thanks for your response. > > Lori A. Harris, HT (ASCP) > Histology Section Leader > GSRMC - Pathology > 3600 NW Samaritan Drive > Corvallis, OR 97330 > 1-541-768-6078 > lharris@samhealth.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 > > > > ------------------------------ > > Message: 24 > Date: Fri, 6 Jan 2006 09:43:24 -0800 (PST) > From: Rene J Buesa > Subject: RE: [Histonet] Processing/embedding question - your opinion > To: Bonnie Whitaker , > histonet@lists.utsouthwestern.edu > Message-ID: <20060106174324.46935.qmail@web61222.mail.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > Bonnie: > That is my personal and very particular opinion, and I think I am > entitled to that, > in the same way that you have expressed yours. > I believe that this is covered by the freedom of expression, is it not? > Ren? J. > > Bonnie Whitaker wrote: > Rene, > > What is "lazy and indolent" about saving processed tissue without > embedding > it? I frequently collect many tissues for embedding in multi-tissue > blocks, > and I see no point in embedding the tissue temporarily while I continue to > collect additional material. It makes no difference if the paraffin > coating > is minimal or a full block. You can also store a lot more control tissue > in > a small area if it is maintained in unembedded cassettes. One cassette can > hold several blocks worth of material. > > Bonnie Whitaker > Lab Manager > Brown & Associates Medical Laboratories > 8076 El Rio > Houston, Texas 77054 > 713-741-6677 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J > Buesa > Sent: Friday, January 06, 2006 10:58 AM > To: foley1; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Processing/embedding question - your opinion > > > Hi Foley: > I have seen that done in several places. I have also seen all the > cassettes wrapped in aluminum foil and kept that way (like a pizza > left-over). > I cannot tell you what would be the consequences for the morphology or > future tests. > Theoretically speaking probably this practice will not affect because the > tissue itself and all its components are supposedly embedded in paraffin > that will just solidify. > I personally do not like this to be done. For me, personally, it indicates > laziness, indolence and even "disrespect" for the tissue sample. > Once I was confronted with the need of keeping processed cassettes in a > secure way before casting the blocks. My solution was to put all the > cassettes in a shallow > plastic container, place all the blocks in it, add melted paraffin and > prepare one single block, as large as the container. When I was able to > prepare the blocks individually, I melted the paraffin and prepared the > blocks. > Hope this will help you! > Ren? J. > > foley1 wrote: > Does anyone routinely allow for the hot wax to drain off multiple > cassettes of processed tissue and be held at room temperature for multiple > days (6 > days) before embedding? What would the consequences be to morphology, > possible immunohistochemistry and molecular (DNA) studies? > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > --------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > --------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less > > ------------------------------ > > Message: 25 > Date: Fri, 6 Jan 2006 11:51:03 -0600 > From: "Allen, Rhonda" > Subject: RE: [Histonet] Processing/embedding question - your opinion > To: "Rene J Buesa" , "Bonnie Whitaker" > , > Message-ID: > > > Content-Type: text/plain; charset="iso-8859-1" > > Renee, > It may be your opinion, but to those of us who have done it out of > necessity, not being "lazy and indolent", it sounds like you are trying to > make us feel "lazy and indolent". > > Rhonda Allen BA HT(ASCP)HTL, QIHC > Histotechnology Specialist II > Stowers Institute > 1000 E. 50th Street > Kansas City, MO 64110 > 816-926-4305 > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J > Buesa > Sent: Friday, January 06, 2006 11:43 AM > To: Bonnie Whitaker; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Processing/embedding question - your opinion > > > Bonnie: > That is my personal and very particular opinion, and I think I am > entitled to that, > in the same way that you have expressed yours. > I believe that this is covered by the freedom of expression, is it not? > Ren? J. > > Bonnie Whitaker wrote: > Rene, > > What is "lazy and indolent" about saving processed tissue without > embedding it? I frequently collect many tissues for embedding in > multi-tissue blocks, and I see no point in embedding the tissue > temporarily while I continue to collect additional material. It makes no > difference if the paraffin coating is minimal or a full block. You can > also store a lot more control tissue in a small area if it is maintained > in unembedded cassettes. One cassette can hold several blocks worth of > material. > > Bonnie Whitaker > Lab Manager > Brown & Associates Medical Laboratories > 8076 El Rio > Houston, Texas 77054 > 713-741-6677 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J > Buesa > Sent: Friday, January 06, 2006 10:58 AM > To: foley1; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Processing/embedding question - your opinion > > > Hi Foley: > I have seen that done in several places. I have also seen all the > cassettes wrapped in aluminum foil and kept that way (like a pizza > left-over). I cannot tell you what would be the consequences for the > morphology or future tests. Theoretically speaking probably this practice > will not affect because the tissue itself and all its components are > supposedly embedded in paraffin that will just solidify. I personally do > not like this to be done. For me, personally, it indicates laziness, > indolence and even "disrespect" for the tissue sample. Once I was > confronted with the need of keeping processed cassettes in a secure way > before casting the blocks. My solution was to put all the cassettes in a > shallow plastic container, place all the blocks in it, add melted paraffin > and prepare one single block, as large as the container. When I was able > to prepare the blocks individually, I melted the paraffin and prepared the > blocks. Hope this will help you! Ren? J. > > foley1 wrote: > Does anyone routinely allow for the hot wax to drain off multiple > cassettes of processed tissue and be held at room temperature for multiple > days (6 > days) before embedding? What would the consequences be to morphology, > possible immunohistochemistry and molecular (DNA) studies? > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > --------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > --------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 26 > Date: Fri, 6 Jan 2006 11:53:08 -0600 > From: "Harrison, Sandra C." > Subject: [Histonet] microwave tissue processing > To: > Message-ID: > <736E8889E98B8F4FBBD29FDFEC8074BA49BF79@VHAV23MSGA2.v23.med.va.gov> > Content-Type: text/plain; charset="us-ascii" > > Hi, > > I'm looking into microwave tissue processors. If you are using one, > please tell me the brand and whether you are pleased with the results. > Do you find the instrument reliable? > > Thanks, Histonetters. > > Sandy > > > > ------------------------------ > > Message: 27 > Date: Fri, 6 Jan 2006 09:54:58 -0800 > From: "Trajkovic, Dusko" > Subject: RE: [Histonet] Processing/embedding question - your opinion > To: > Message-ID: > <3AD0BD3142459B4E9B12CBEAFF2B89B2D896AB@lajamrexm01.amer.pfizer.com> > Content-Type: text/plain; charset="iso-8859-1" > > I have yet to see any published information stating that un-embedded > tissue left to solidify, is damaging in any way or form. My colleagues an > I in research, have done it numerous times, for various reasons, non were > for being lazy. > > Dusko Trajkovic > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J > Buesa > Sent: Friday, January 06, 2006 9:43 AM > To: Bonnie Whitaker; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Processing/embedding question - your opinion > > Bonnie: > That is my personal and very particular opinion, and I think I am > entitled to that, > in the same way that you have expressed yours. > I believe that this is covered by the freedom of expression, is it not? > Ren? J. > > Bonnie Whitaker wrote: > Rene, > > What is "lazy and indolent" about saving processed tissue without > embedding > it? I frequently collect many tissues for embedding in multi-tissue > blocks, > and I see no point in embedding the tissue temporarily while I continue to > collect additional material. It makes no difference if the paraffin > coating > is minimal or a full block. You can also store a lot more control tissue > in > a small area if it is maintained in unembedded cassettes. One cassette can > hold several blocks worth of material. > > Bonnie Whitaker > Lab Manager > Brown & Associates Medical Laboratories > 8076 El Rio > Houston, Texas 77054 > 713-741-6677 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J > Buesa > Sent: Friday, January 06, 2006 10:58 AM > To: foley1; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Processing/embedding question - your opinion > > > Hi Foley: > I have seen that done in several places. I have also seen all the > cassettes wrapped in aluminum foil and kept that way (like a pizza > left-over). > I cannot tell you what would be the consequences for the morphology or > future tests. > Theoretically speaking probably this practice will not affect because the > tissue itself and all its components are supposedly embedded in paraffin > that will just solidify. > I personally do not like this to be done. For me, personally, it indicates > laziness, indolence and even "disrespect" for the tissue sample. > Once I was confronted with the need of keeping processed cassettes in a > secure way before casting the blocks. My solution was to put all the > cassettes in a shallow > plastic container, place all the blocks in it, add melted paraffin and > prepare one single block, as large as the container. When I was able to > prepare the blocks individually, I melted the paraffin and prepared the > blocks. > Hope this will help you! > Ren? J. > > foley1 wrote: > Does anyone routinely allow for the hot wax to drain off multiple > cassettes of processed tissue and be held at room temperature for multiple > days (6 > days) before embedding? What would the consequences be to morphology, > possible immunohistochemistry and molecular (DNA) studies? > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > --------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > --------------------------------- > Yahoo! DSL Something to write home about. Just $16.99/mo. or less > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ---------------------------------------------------------------------- > LEGAL NOTICE > Unless expressly stated otherwise, this message is confidential and may be > privileged. It is intended for the addressee(s) only. Access to this > E-mail by anyone else is unauthorized. If you are not an addressee, any > disclosure or copying of the contents of this E-mail or any action taken > (or not taken) in reliance on it is unauthorized and may be unlawful. If > you are not an addressee, please inform the sender immediately. > > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 26, Issue 4 > *************************************** > From gcallis <@t> montana.edu Fri Jan 6 18:07:37 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Jan 6 18:07:50 2006 Subject: [Histonet] Microwave drying/paraffin sections from Histonet archives - long but maybe pertinent? Message-ID: <6.0.0.22.1.20060106160758.01b4cc10@gemini.msu.montana.edu> If glass could heated up by microwaves, then why are we able to remove pyrex beakers, coplin jars or in our homes, glass pyrex measuring cups containing water heated to visual boiling with little skin protection i.e. bare hands (although I don't advocate this in home or laboratory, a hot mitt is always a good idea!). However the longer it takes to heat the water in my kitchen MW, the hotter the glass becomes over time with the transmitted heat. Some glass containers have gold borders or other components(?) that are not microwaveable, with warnings - but these are not glass slides. The question and discussions on MW drying are numerous, but three messages stood out and should be considered if wanting to do this. The biggest concern should be what kind of artifact or tissue section damage is created with MW drying? Histotechnicians have experienced this, per archive messaging. Although MW drying saves time, does it lead to eventual problems in staining, diagnosis, etc.? We tend to dry our slides at lower temperatures in an oven i.e. Buessa convection oven method. I thought these excerpts from a Peggy Wenk message about microwaving pertinent, and the way I always understood a MW to work. Microwaves cause molecules that have a charge (like water or alcohol) to align to the wave as it passes. After the wave passes, the charged molecule goes back to whatever direction it wants to point, at least until the next wave passes by, at which point the charged molecule realigns again. So, you have a bunch of charged molecules aligning and becoming randomly arranged, millions of times a second (frequency of a microwave). While these charged molecules are aligning and un-aligning, they are bumping into each other. This causes friction, which causes heat. Which is how aqueous or alcoholic solutions (or your supper) heats up. Now, some things do NOT have charges to them. Like the glass or pyrex or plastic in the coplin jar or casserole dish. So the molecule do NOT move. So there is NO friction. Therefore no warming. (The only exception, if you want to consider this, is drying slides in the microwave oven. But in this case, the water between the slide and the tissue/paraffin section is being heated. In this case, it is usually advised to have a beaker of water in the back corner of the microwave, so that there is something to absorb the microwaves, so that the oven is not harmed.) Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 On slide drying, there may be some other effects you do not want, like introducing artifact to the tissue sections - Charles.Embrey[SMTP:Charles.Embrey@carle.com] Sent: Wednesday, October 10, 2001 3:43 PM I have worked at places before that dried slides in the microwave. In my own lab I will not. The microwave tends to overheat during this process and I saw several burn up. I was always afraid of fire. Drying slides in the microwave also creates interesting artifacts when the water boils under the tissue. I just don't like it. I am sure however that others do and will probably beat me up over my stand but it's just my personal preference and experienced opinion. No daggers please Charles R. Embrey Jr., PA(AAPA), HT(ASCP) Histology Supervisor Carle Clinic Urbana, IL And from a Microwaving expert: Subject: Drying paraffin slides Date: Tue, 02 Apr 2002 12:01:43 -0600 Hi - Just a point of terminology - whether heating slides in an oven, with forced air, or in the microwave, you are NOT drying the paraffin. You are drying the layer of water between the paraffin and the slide. Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From katri <@t> cogeco.ca Sat Jan 7 22:17:53 2006 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Sat Jan 7 22:18:00 2006 Subject: [Histonet] Bone marrow aspirate smears Message-ID: <000701c6140a$7eba39c0$6a9a9618@Katri> Hi Histonetters, This is a bit outside of histology, but I'll try for help anyway. A research colleague of mine is wondering, how to best collect bone marrow aspirates for making smears or cytospins for immunocytochemistry. What if the marrow collection is done in one place and have to be transported to place of testing? Any ideas, who would have general information about this type of specimen? Katri Katri Tuomala Hamilton, Ontario, Canada From rjbuesa <@t> yahoo.com Sun Jan 8 07:43:32 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sun Jan 8 07:43:42 2006 Subject: [Histonet] Bone marrow aspirate smears In-Reply-To: <000701c6140a$7eba39c0$6a9a9618@Katri> Message-ID: <20060108134332.14725.qmail@web61218.mail.yahoo.com> Hi Kari: In our hospital the residents sample the bone marrow and prepare the smears in site and place them in regular slides folders.The aspirates are fixed in neutral buffered formalin and both are brought to the histology lab. The smears (already air dried ) are fixed with methanol as any blood smear and 2 are stained (Wright-Giemsa and Perl's for iron) and the rest saved for any further test (like IHC). The aspirates are filtered and a block is prepared for histology. The small bone fragment is decalcified with EDTA (only) for 4-6 hours, and processed afterwards. This protocol has worked well for over 20 years. Hope this will help! Ren? J. Katri Tuomala wrote: Hi Histonetters, This is a bit outside of histology, but I'll try for help anyway. A research colleague of mine is wondering, how to best collect bone marrow aspirates for making smears or cytospins for immunocytochemistry. What if the marrow collection is done in one place and have to be transported to place of testing? Any ideas, who would have general information about this type of specimen? Katri Katri Tuomala Hamilton, Ontario, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From lpwenk <@t> sbcglobal.net Sun Jan 8 09:12:49 2006 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Sun Jan 8 09:13:02 2006 Subject: [Histonet] HTL certification In-Reply-To: <9AEEF1FB6254224AA355ED285F84916515A37AA7@EXCHVS2.medctr.ad.wfubmc.edu> Message-ID: In the HT/HTL Examination Content Guideline, sent to all applicants and available on the ASCP webpage, it does state the competency differences between a technician and a technologist, and then has an outline, with the "HTL only" marked with " * ". http://www.ascp.org/bor/application/content/index.asp So, looking at the ASCP HT/HTL competencies lists, the HTL candidate is responsible for everything the HT will be tested on, PLUS (and I'm listing only some): - more problem-solving and troubleshooting - causes of discrepancies of tests - abnormal test results due to pathological states - statistics - understandes and enforces safety regulations - specialized tests - investigates and implementes new procedures/equipment - develops lab manuals, reports, guidelines, research protocol - provides instruction in theory, technical skills, safety protocols - provides continuing education - evaluate effectiveness of education programs - management theory, economic impact, management functions - establish technical and administrative procedures, QC, QA, safety and waste management procedures, information management, cost effective measurs, supervision These are some of the additional areas that the HTL exam can test the HTL candidate on (compared to the HT candidate). When we look at the HT/HTL content outline, the "*" that the HTL are tested on (but not HT) also include: - pathology related to fixation and staining - chemistry/biochemistry of fixation, processing, staining - glycol methacrylate sectioning - enzyme staining - IHC staining - refractive index of mounting procedures - management theories and procedures - education theory and procedures - regulations (federal government and accrediting agencies) The Reading List from ASCP indicates the following additional books for HTL: - Bancroft and Gamble - two books on management http://www.ascp.org/bor/application/reading/index.asp So, yes, HTL exam IS harder than HT - enzymes, plastics, IHC, management, education, lot more problem solving and troubleshooting. Yes, more studying is needed. More books are needed. NSH does have a more complete outline of what needs to be studied. It's free to NSH members - not available to non-NSH members. It's based on the ASCP outline, but instead of 2 pages, it's about 12. There are also several self-assessment booklet available through NSH, that would be helpful for HTL candidates: - Laboratory Safety (#14) - Laboratory Operations (# 11) - IHC, enzyme histochemistry, flow cytometer, ISH, EM (#5) http://www.nsh.org/education/materials.html So, yes, all these things are on the HTL exam. Experience requirement to take the HTL exam requires one year full-time experience in the following areas: - Fixation - Processing - Microtomy - Routine and Special Staining This experience could be in only histology, only IHC, only EM, or any combination. It could be human or animal or plant. It could be clinical or veterinary or research. It could be hard tissue or soft tissue. It could be brain only. It could be FS only. ASCP does no specify WHERE the experience is, or which type of tissue or which type of embedding media or which stains. Just that it must include all four areas. However, the written and practical exams will cover all the areas that were mentioned in the outline and competencies. (For example, the 2006 HTL practical exam includes a pan-cytokeratin on a cervix (to include endo- and ectocervix).) So your HTL candidates DO need to know IHC, enzymes, management, etc., even if they are not doing it in their lab. The same as someone working in, say, a derm lab, who even though in their lab they work with just skins and only do 4 special stains - they must be able to identify all tissues and know the theory of all the stains, even if they don't do it in their lab. Passing the ASCP HT and HTL exams indicates a level of knowledge/competency to be able to go work in ANY lab in the country, NOT just the lab that the candidate is currently working at. Hope that helps your candidates. Good luck to them. Peggy A. Wenk, HTL(ASCP)SLS Schools of HT and HTL William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hermina Borgerink Sent: Friday, January 06, 2006 12:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HTL certification Two of my techs will be taking the HTL certification this year. I checked the requirements on the ASCP Board of Registry home page and found that they needed experience in the following areas: fixation, processing, microtomy, and routine as well as special staining. I just wanted to double check with those of you who have recently taken their Board Certifications that EM and ISH experience are no longer required. When I did my HTL certification there still were questions pertaining to both topics, and I just want to make sure they won't be faced with questions they are not trained in. Any response would be greatly appreciated and I thank you in advance. Hermina Hermina M. Borgerink, BA, HT(ASCP)HTL, QIHC Wake Forest University Health Sciences Department of Pathology Medical Center Blvd. Winston-Salem, NC 27157 Tel. (336) 716-1538 Fax. (336) 716-1515 e-mail hborgeri@wfubmc.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Sun Jan 8 11:17:56 2006 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Sun Jan 8 11:22:00 2006 Subject: [Histonet] Microwave slide drying vs. conventional slide dryi ng Message-ID: <5D2189E74151CC42BEC02906BA8996322B914F@exchsrv01.barrynet.barry.edu> I have always left my slides on a 50 degree warming plate for 48 hours. The slides are dry in an hour, but sections will fall off during a PAS reaction. After 48 hr I rarely lose a section no matter what I do to it. 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 Kinsley, David Sent: Friday, January 06, 2006 2:57 PM To: 'Laurie Colbert'; lharris@samhealth.org; Histonet (E-mail) Subject: RE: [Histonet] Microwave slide drying vs. conventional slide dryi ng Can you let me know what tissues samples and temperature you typically use with the hot plate method? I work with animal tissues and usually place my slides on a hot plate at about 45-50 degrees to flatten the sections out (about 1 hour) and then follow by heating them in a 60 degree oven for 4 hours to overnight depending on the sample. Dave -----Original Message----- From: Laurie Colbert [mailto:laurie.colbert@huntingtonhospital.com] Sent: Friday, January 06, 2006 12:38 PM To: lharris@samhealth.org; Histonet (E-mail) Subject: RE: [Histonet] Microwave slide drying vs. conventional slide drying Lori, We don't use either. We simply put our drained slides on the hot plate for about 5 minutes. In previous jobs, we always used an oven, and when I came here I was really nervous about just using the hot plate - but it works! Laurie Colbert Huntington Hospital Pasadena, CA -----Original Message----- From: lharris@samhealth.org [mailto:lharris@samhealth.org] Sent: Friday, January 06, 2006 8:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave slide drying vs. conventional slide drying We are having a discussion about slide drying at out facility. How many of you out there use a microwave to dry your H&E slides before staining versus using a conventional 70 degree Celsius hot air slide drying oven for 20 minutes? I personally against microwave slide drying but the doctors like it because it takes less time (3 min. 30 sec. on medium setting). Thanks for your response. Lori A. Harris, HT (ASCP) Histology Section Leader GSRMC - Pathology 3600 NW Samaritan Drive Corvallis, OR 97330 1-541-768-6078 lharris@samhealth.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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. _______________________________________________ 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 pruegg <@t> ihctech.net Sun Jan 8 11:58:26 2006 From: pruegg <@t> ihctech.net (patsy ruegg) Date: Sun Jan 8 11:58:54 2006 Subject: [Histonet] Bone marrow aspirate smears In-Reply-To: <20060108134332.14725.qmail@web61218.mail.yahoo.com> Message-ID: <200601081758.k08HwQZj004704@pro12.abac.com> We assisted the docs doing the bm biopsies. We provided a anti-coag.(edta or heprine) lined tube to collect the aspirate in, then carried it back to the lab and made the smear preps and cell blocks for histology blocks in the lab, I found it more efficient to do in the lab than at the patients bedside, we tried to get in and out of the patients way asap. 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 Rene J Buesa Sent: Sunday, January 08, 2006 6:44 AM To: Katri Tuomala; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Bone marrow aspirate smears Hi Kari: In our hospital the residents sample the bone marrow and prepare the smears in site and place them in regular slides folders.The aspirates are fixed in neutral buffered formalin and both are brought to the histology lab. The smears (already air dried ) are fixed with methanol as any blood smear and 2 are stained (Wright-Giemsa and Perl's for iron) and the rest saved for any further test (like IHC). The aspirates are filtered and a block is prepared for histology. The small bone fragment is decalcified with EDTA (only) for 4-6 hours, and processed afterwards. This protocol has worked well for over 20 years. Hope this will help! Ren? J. Katri Tuomala wrote: Hi Histonetters, This is a bit outside of histology, but I'll try for help anyway. A research colleague of mine is wondering, how to best collect bone marrow aspirates for making smears or cytospins for immunocytochemistry. What if the marrow collection is done in one place and have to be transported to place of testing? Any ideas, who would have general information about this type of specimen? Katri Katri Tuomala Hamilton, Ontario, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From beingmary53 <@t> sbcglobal.net Sun Jan 8 17:37:55 2006 From: beingmary53 <@t> sbcglobal.net (MARY JOHNSON) Date: Sun Jan 8 17:38:05 2006 Subject: [Histonet] formalin pigment Message-ID: <20060108233755.73413.qmail@web81604.mail.mud.yahoo.com> hi histonette, I am having problems with formalin pigments, I am sure it is because of the out dated Formaldehyde that the moruge is using. Can any one help me prove this what test can i use? Is the paper pH ok to prove this. I need help with this one From rjbuesa <@t> yahoo.com Mon Jan 9 08:22:20 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Jan 9 08:22:31 2006 Subject: [Histonet] formalin pigment In-Reply-To: <20060108233755.73413.qmail@web81604.mail.mud.yahoo.com> Message-ID: <20060109142220.72713.qmail@web61211.mail.yahoo.com> Mary: Formalin pigment, also known as "hematin", has been found in tissues stored in formalin for long periods of time since formalin began to be used, Hematin which, as you know, is a dark brown, microcrystaline and iron-negative pigment also develops in Bouin's fixed tissue, specially in spleen, heart and almost any blood rich tissue. It was so frequent years ago than it lead to the development of neutral buffered formalin (NBF) or formalin containing enough salts to allow the pH to remain neutral (buffered) after long periods of time. You can use a pH indicator paper strip because you only want to determine if the formalin is acid (less than pH 7) or not. You don't want to determine the exact pH for which you will need a calibrated pH-meter. On the other hand there are good pH paper strips like one called "pHydrion Vivid 6-8" that will tell you the pH from values of 6.0 to 8.0 in increments of 0.2 to 0.4 units. Another pH strips called "color pHast" determine pH range from "0" to "14" in pH 1 unit increments. I hope this will help you! Ren? J. MARY JOHNSON wrote: hi histonette, I am having problems with formalin pigments, I am sure it is because of the out dated Formaldehyde that the moruge is using. Can any one help me prove this what test can i use? Is the paper pH ok to prove this. I need help with this one _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos ? Showcase holiday pictures in hardcover Photo Books. You design it and we?ll bind it! From vd38 <@t> georgetown.edu Mon Jan 9 09:42:15 2006 From: vd38 <@t> georgetown.edu (Vernon Dailey) Date: Mon Jan 9 09:42:31 2006 Subject: [Histonet] X-Gal staining Message-ID: <43C28457.20006@georgetown.edu> Hi Histonet, I am interested in running IHC on frozen and FFPE mouse tissues for B-Galactosidase. Can anyone recommend a vendor and product # for a primary towards B-Gal that has been successful on **tissue**. Also, could one stain for B-Gal after/before performing an X-Gal reaction? I am interested in colocalizing several other Abs with B-Gal so IHC would be helpful above the X-Gal. Thanks, Vernon Dailey Georgetown University From sa.drew <@t> hosp.wisc.edu Mon Jan 9 09:46:06 2006 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Mon Jan 9 09:46:17 2006 Subject: [Histonet] Bartonella henselae Message-ID: Is anyone aware of where one of our pathologists might get immunostains done with a monoclonal antibody to Bartonella henselae? Thank you! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 From gcallis <@t> montana.edu Mon Jan 9 09:46:27 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Jan 9 09:46:35 2006 Subject: [Histonet] Microwave drying/paraffin sections from Histonet archives -long but maybe pertinent? In-Reply-To: <43C0ADB6.CEB87DCF@uwo.ca> References: <6.0.0.22.1.20060106160758.01b4cc10@gemini.msu.montana.edu> <43C0ADB6.CEB87DCF@uwo.ca> Message-ID: <6.0.0.22.1.20060109084008.01b6bf10@gemini.msu.montana.edu> John, I agree and that is why some glass and ceramics have warnings on them to NOT microwave. That was the point of this comment in the text of my reply. I just did not say ceramics, but I have some in my home that are forbidden in a MW oven. I am aware of warnings - at least with my home MW oven - that MW'ing dry containers in a MW or running a MW without water present is not good for the MW oven either. "Some glass containers have gold borders or other components(?) that are not microwaveable, with warnings - but these are not glass slides." At 11:14 PM 1/7/2006, you wrote: >Dear Gayle, > >Food for thought follows. > >My subjective feeling is that ceramics are heated >by microwaves. The whole plate gets very hot, not >just the area on which the watery and fatty foods >were resting. > >John Kiernan >London, Canada. >___________________________________________________ >Gayle Callis wrote: Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From jladuc <@t> trudeauinstitute.org Mon Jan 9 10:15:37 2006 From: jladuc <@t> trudeauinstitute.org (Judith LaDuc) Date: Mon Jan 9 10:10:18 2006 Subject: [Histonet] NYS Licensure Message-ID: To view the Clinical Laboratory Technology Act and read the entire bill, which does include histotechnicians and histotechnologists, you may use this link: http://www.op.nysed.gov/article165.htm To check for updates as they become available, use this link: http://www.op.nysed.gov/clp.htm I spoke with Dr. Kathleen Doyle today at the NYS Education Department. They are putting the regulations/requirements for licensure together. It is their intent to make them generic acording to practice and as broad as possible. The same is to be said of the exams. They will be exams of practice, not education, it is up to schools to test your education. She stated an example of Physician licensure in NYS. An MD can be licensed to practice medicine in NYS but an employer may require additional board certification specialties. NYS will not test for those. Currently, as they look at grandparenting they will be looking for five of more years of experience to qualify for this. You will be required to submit attestations from former work experience and there is no time limit. Again, they are trying to make this fairly broad. As to where you have attained this experience, she said it could be from Brazil to Connecticut. One of the reasons that you don't hear frequent updates is that it is a fairly lengthy process. Here is what they are trying to use for a timeline at the moment: 1. Board of Directors are really pushing to get the education regulations done in the next two months 2. Submit language to the NYS Register for 40 to 60 day review 3. Submit to Board of Regents where it is hopefully enacted 4. Finish registration materials 5. Post registration on: http://www.op.nysed.gov/clp.htm 6. Begin processing applications 7. If too close to September 1, 2006, may have to ask for extension of date so that professional will not be non-compliant with law Note: Dr. Doyle stated that you would have up to two years to apply. You would not be able to work during that time, but there was a window of opportunity there. (Sorry, I can't remember if this referred to grandparenting or not. I was trying to take notes). So keep checking the clp.htm site and try to join us in Saratoga Springs in April :) Judy LaDuc, President NYS Histotechnological Society From histology.bc <@t> shaw.ca Mon Jan 9 10:10:37 2006 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Mon Jan 9 10:11:39 2006 Subject: [Histonet] formalin pigment In-Reply-To: <20060108233755.73413.qmail@web81604.mail.mud.yahoo.com> References: <20060108233755.73413.qmail@web81604.mail.mud.yahoo.com> Message-ID: <43C28AFD.6070008@shaw.ca> The situation you are decribing, with old solutions of formaldehyde, long storage times ususally associated with morgue specimens, is a classical formalin pigment-producing scenario. To confirm that the pigment is formalin pigment (correctly known as acid formol hematin) just examine a section containing the pigment using a polarized light microscope. Formalin pigment is birefringent and will appear bright on a dark background. No other common pigments are birefringent. Also if it tends to be more concentrated in areas where there are numerous red blood cells, in blood vessels, in spleen, etc., you can be pretty sure you are dealing with formalin pigment. To get rid of the pigment from the sections, treat the de-waxed sections with saturated alcoholic picric acid for about 30 minutes, then wash the sections very well to remove the yellow staining, then stain as usual. Paul Bradbury Kamloops, BC, Canada MARY JOHNSON wrote: >hi histonette, I am having problems with formalin pigments, I am sure it is because of the out dated Formaldehyde that the moruge is using. Can any one help me prove this > what test can i use? Is the paper pH ok to prove this. I need help with this one >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From atg <@t> med.auth.gr Mon Jan 9 10:10:11 2006 From: atg <@t> med.auth.gr (Aliki Xohelli) Date: Mon Jan 9 10:23:40 2006 Subject: [Histonet] (no subject) Message-ID: <1136823011.43c28ae375fd8@webmail.med.auth.gr> hi , i would like to ask you if you know which is the best procedure as far as bone marrow sections is concerned. I am going to use adult male wistar rats. Thank you in advance Alice Xohelli From Tbarnhart <@t> primecare.org Mon Jan 9 10:37:24 2006 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Mon Jan 9 10:39:12 2006 Subject: [Histonet] Microwave drying of slides Message-ID: <1779904B5E82D511914C00D0B793339205BFDAD5@exchangent> I have been drying slides in a microwave purchased from our local retail store for 10 plus years. We have no problems with morphology, sections lifting or broken slides. One minute, twenty seconds on 3/4 power. Allow to sit in the microwave an additional two minutes, 40 seconds (four minutes total). Lay the slides on the counter to cool for a few seconds and load in the stainer. Our microwave is a small 600w with a revolving tray. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. From pmarcum <@t> vet.upenn.edu Mon Jan 9 10:41:17 2006 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Mon Jan 9 10:41:32 2006 Subject: [Histonet] formalin pigment In-Reply-To: <43C28AFD.6070008@shaw.ca> References: <20060108233755.73413.qmail@web81604.mail.mud.yahoo.com> <43C28AFD.6070008@shaw.ca> Message-ID: <6.1.1.1.2.20060109113143.01a26870@mail.vet.upenn.edu> Paul is absolutely correct and a great explanation of the way to define formalin pigment quickly. It is always best to be sure they have fresh formalin in the morgue and if possible with tissues of interest to change the solution within a reasonable time frame of a minimum of once a month. Brain tissue particularly will be fixed better and more completely if the formalin is changes several times in the first one to two weeks of fixation as the autolysis occurring naturally changes the pH. This can cause the buffering salts to form a crust around the outer areas. This will prevent penetration of the fixative and cause poorly fixed center of the brain. I have also used the old standard of an iodine step followed by sodium thiosulfate solution at the end of my deparaffinization step with good rinsing prior to staining to remove pigment. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu At 11:10 AM 1/9/2006, Paul Bradbury wrote: >The situation you are decribing, with old solutions of formaldehyde, long >storage times ususally associated with morgue specimens, is a classical >formalin pigment-producing scenario. > >To confirm that the pigment is formalin pigment (correctly known as acid >formol hematin) just examine a section containing the pigment using a >polarized light microscope. Formalin pigment is birefringent and will >appear bright on a dark background. No other common pigments are >birefringent. Also if it tends to be more concentrated in areas where >there are numerous red blood cells, in blood vessels, in spleen, etc., you >can be pretty sure you are dealing with formalin pigment. > >To get rid of the pigment from the sections, treat the de-waxed sections >with saturated alcoholic picric acid for about 30 minutes, then wash the >sections very well to remove the yellow staining, then stain as usual. > >Paul Bradbury >Kamloops, BC, Canada > > >MARY JOHNSON wrote: > >>hi histonette, I am having problems with formalin pigments, I am sure it >>is because of the out dated Formaldehyde that the moruge is using. Can >>any one help me prove this what test can i use? Is the paper pH ok to >>prove this. I need help with this one >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cbass <@t> bidmc.harvard.edu Mon Jan 9 10:48:03 2006 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Mon Jan 9 10:48:13 2006 Subject: [Histonet] neurite visualization in cell culture Message-ID: Hello, I am doing a neurite outgrowth assay in vitro and I need to add some contrast in order to visualize the neurites. Is there a protocol for staining entire cells that will bring out this detail? I just need to darken the entire cell and the neurite. I have heard of some people using commassie blue for this purpose. Does anyone have a protocol for this? Thanks, Caroline Bass From Sandra.Harrison3 <@t> va.gov Mon Jan 9 11:24:57 2006 From: Sandra.Harrison3 <@t> va.gov (Harrison, Sandra C.) Date: Mon Jan 9 11:25:09 2006 Subject: [Histonet] John Kiernan's textbook Message-ID: <736E8889E98B8F4FBBD29FDFEC8074BA49BF80@VHAV23MSGA2.v23.med.va.gov> Hi Histonetters, I'm wondering if anyone can tell me who I would contact to purchase Histological and Histochemical Techniques: Theory and Practice by John Tiernan, the 3rd Edition. Thanks for the responses to my questions about microwave tissue processors. This board is a great resource. Sandy From Janet.Keeping <@t> cna.nl.ca Mon Jan 9 11:38:24 2006 From: Janet.Keeping <@t> cna.nl.ca (Keeping, Janet) Date: Mon Jan 9 11:38:37 2006 Subject: [Histonet] John Kiernan's book Message-ID: My copy says distributed in the USA by Oxford University Press INC. 198 Madison Avenue, New York ,NY 10016 Janet Keeping, A.R.T., B.Voc.Ed. Instructor Medical Laboratory Sciences 1 Prince Philip Drive P.O. Box 1693, St. John's NL, Canada, A1C 5P7 709 758-7657 tel 709 758-7635 fax From nienhuis <@t> ucla.edu Mon Jan 9 12:38:23 2006 From: nienhuis <@t> ucla.edu (nienhuis@ucla.edu) Date: Mon Jan 9 12:38:32 2006 Subject: [Histonet] Tract tracing with HRP or CTB In-Reply-To: <20060108233755.73413.qmail@web81604.mail.mud.yahoo.com> References: <20060108233755.73413.qmail@web81604.mail.mud.yahoo.com> Message-ID: <20060109103823.zy5sjafy0w0k40k4@mail.ucla.edu> Has anybody done tract tracing with WGA-HRP or cholera toxin? We have done it before, but have always processed the tissue soon after the animal was sac'd. We may need to wait for a short period before processing the brains. Does anybody have any suggestions for avoiding loss of signal, or the best way to store this tissue? Or is it even a problem? Bob Nienhuis UCLA / VA Med Center nienhuis@ucla.edu From dpconsult <@t> earthlink.net Mon Jan 9 13:05:58 2006 From: dpconsult <@t> earthlink.net (Dick Paulson [Source Medical Products]) Date: Mon Jan 9 13:06:15 2006 Subject: [Histonet] John Kiernan's textbook Message-ID: Sandy, You can find it on the web at www.Amazon.com Here is the direct link to the book: http://www.amazon.com/gp/product/0750649364/ref=ed_oe_p/103-4055781-9131043? %5Fencoding=UTF8 Dick Paulson Hi Histonetters, I'm wondering if anyone can tell me who I would contact to purchase Histological and Histochemical Techniques: Theory and Practice by John Kiernan, the 3rd Edition. Thanks for the responses to my questions about microwave tissue processors. This board is a great resource. Sandy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From stamptrain <@t> yahoo.com Mon Jan 9 13:10:33 2006 From: stamptrain <@t> yahoo.com (Roger Moretz) Date: Mon Jan 9 13:10:45 2006 Subject: [Histonet] Ki-67 or PCNA on FFPE primate tissue Message-ID: <20060109191033.24368.qmail@web50304.mail.yahoo.com> Anyone with experience using any of the available antibodies for FFPE primate tissue? Any preferences? Roger Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals,Inc. Ridgefield, CT __________________________________________ Yahoo! DSL ? Something to write home about. Just $16.99/mo. or less. dsl.yahoo.com From DNolan <@t> evanhospital.com Mon Jan 9 13:19:52 2006 From: DNolan <@t> evanhospital.com (Donna M. Nolan) Date: Mon Jan 9 13:22:09 2006 Subject: [Histonet] Leica microtome for sale Message-ID: <2C7B7A4D7DE62A46BC2EFB70D3ABF9326A49B0@EVANXSCL.evanhospital.net> We have a Leica 1512 microtome with standard knife holder we no longer use. Is any one interested in purchasing this from us or know of a company that purchases used equipment? Donna Nolan Evangelical Hospital Lewisburg PA 17837 From jkiernan <@t> uwo.ca Mon Jan 9 14:56:24 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Mon Jan 9 14:56:35 2006 Subject: [Histonet] John Kiernan's textbook References: <736E8889E98B8F4FBBD29FDFEC8074BA49BF80@VHAV23MSGA2.v23.med.va.gov> Message-ID: <43C2CDF8.E2D251B4@uwo.ca> The publisher has been changed for a second time, and is now Scion Publishing Ltd, Bloxham, Oxfordshire, UK. For information: http://www.scionpublishing.com/pub/pub_result.php?keywords=Kiernan To the best of my knowledge the book is still distributed in N. America by Oxford University Press: USA: http://www.us.oup.com/us/catalog/general/subject/Medicine/LaboratoryScience/?view=usa&ci=0750649364 Canada: http://www.oup.com/ca/isbn/0-7506-4936-4 -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Harrison, Sandra C." wrote: > > Hi Histonetters, > > > > I'm wondering if anyone can tell me who I would contact to purchase > Histological and Histochemical Techniques: Theory and Practice by John > Tiernan, the 3rd Edition. > > > > Thanks for the responses to my questions about microwave tissue > processors. This board is a great resource. > > > > Sandy > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Mon Jan 9 15:05:23 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Mon Jan 9 15:05:28 2006 Subject: [Histonet] neurite visualization in cell culture References: Message-ID: <43C2D013.2E4D4015@uwo.ca> Carri & Ebendal 1989 Coomassie brilliant blue R250 (CI 42660): 0.1g Methanol: 25 ml Acetic acid: 10 ml Water to 100 ml Stain for 20-30 minutes. For details see the original paper in Stain Technology 64:50-52. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Caroline Bass wrote: > > Hello, > > I am doing a neurite outgrowth assay in vitro and I need to add some > contrast in order to visualize the neurites. Is there a protocol for > staining entire cells that will bring out this detail? I just need > to darken the entire cell and the neurite. I have heard of some > people using commassie blue for this purpose. Does anyone have a > protocol for this? > > Thanks, > > Caroline Bass From JMacDonald <@t> mtsac.edu Mon Jan 9 15:34:00 2006 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Mon Jan 9 15:33:56 2006 Subject: [Histonet] formalin pigment In-Reply-To: <6.1.1.1.2.20060109113143.01a26870@mail.vet.upenn.edu> Message-ID: Mercury pigment is also birefringent, and is removed by the iodine/sodium thiosulfate treatment. Formalin pigment is removed by the picric acid treatment that Paul mentioned as well as by alkaline alcohol. Formalin pigment will form in bloody tissues in the presence of an acid environment. You will see it in the blood vessels and where blood has accumulated. In the kidney it tends to accumulate in the glomeruli. Jennifer MacDonald Pamela Marcum Sent by: histonet-bounces@lists.utsouthwestern.edu 01/09/2006 08:41 AM To Paul Bradbury , MARY JOHNSON , HistoNet Server cc Subject Re: [Histonet] formalin pigment Paul is absolutely correct and a great explanation of the way to define formalin pigment quickly. It is always best to be sure they have fresh formalin in the morgue and if possible with tissues of interest to change the solution within a reasonable time frame of a minimum of once a month. Brain tissue particularly will be fixed better and more completely if the formalin is changes several times in the first one to two weeks of fixation as the autolysis occurring naturally changes the pH. This can cause the buffering salts to form a crust around the outer areas. This will prevent penetration of the fixative and cause poorly fixed center of the brain. I have also used the old standard of an iodine step followed by sodium thiosulfate solution at the end of my deparaffinization step with good rinsing prior to staining to remove pigment. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu At 11:10 AM 1/9/2006, Paul Bradbury wrote: >The situation you are decribing, with old solutions of formaldehyde, long >storage times ususally associated with morgue specimens, is a classical >formalin pigment-producing scenario. > >To confirm that the pigment is formalin pigment (correctly known as acid >formol hematin) just examine a section containing the pigment using a >polarized light microscope. Formalin pigment is birefringent and will >appear bright on a dark background. No other common pigments are >birefringent. Also if it tends to be more concentrated in areas where >there are numerous red blood cells, in blood vessels, in spleen, etc., you >can be pretty sure you are dealing with formalin pigment. > >To get rid of the pigment from the sections, treat the de-waxed sections >with saturated alcoholic picric acid for about 30 minutes, then wash the >sections very well to remove the yellow staining, then stain as usual. > >Paul Bradbury >Kamloops, BC, Canada > > >MARY JOHNSON wrote: > >>hi histonette, I am having problems with formalin pigments, I am sure it >>is because of the out dated Formaldehyde that the moruge is using. Can >>any one help me prove this what test can i use? Is the paper pH ok to >>prove this. I need help with this one >>_______________________________________________ >>Histonet mailing list >>Histonet@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 Vfierke <@t> SNBLUSA.com Mon Jan 9 17:53:04 2006 From: Vfierke <@t> SNBLUSA.com (Vaughn Fierke) Date: Mon Jan 9 17:53:26 2006 Subject: [Histonet] histotechnology program everett washington Message-ID: <3CB1E2E5EC8E9C48A06C8E0967EB82B20E5D0A@MAIL01.snblusa.com> Finally, the West Coast is finally getting ready to launch a new formal histotechnology program at Cogswell College in Everett, Washington. It's first class, An Introduction to Histology Technique I, is currently accepting students for credit or to audit. The first class will start Tuesday 1/24. The class will be a 2 hour lecture series; a lab on Thursday will be 4 hours. The sessions will last through April. Upon completion, the successful student can be eligible to begin attending a clinical/research lab for practical experience and credit. Other related science classes are also available. This new program is intended to be a 2-year (AA with HT eligibility) and 4-year degree in histotechnology with eligibility to take the HTL-level certification. The program is being evaluated to be approved as an accredited program through the state's education board and for all successful students to meet the requirements for ASCP-certifications. Initial approval processes are going very well. Those in the education fields know well how difficult it is in acquiring equipment and supplies in the initial phases. I would like to make a request for any labs to help in developing this initial process in donating at least one item from your stores or just cleaning out the lab closet and forwarding them to the college lab. Please contact me for more information on the program and anything you would like to contribute. Thank you to the area labs and vendors for your support already. Vaughn Fierke Histology Supervisor 425-322-2479 From Lynne.Bell <@t> hitchcock.org Tue Jan 10 07:22:21 2006 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Tue Jan 10 07:22:31 2006 Subject: [Histonet] Stains for pneumocystis Message-ID: My fellow Histonetters, I would like to know what stain you are using for pneumocystis. We are presently using a "modified" GMS that takes approximately one hour and 15 minutes to perform. A few of our physicians would like something a bit faster. (Of course, I would like to get the specimen sometimes before 3 p.m., but that is another story for another time!) I have also found that a Thin Prep slide from a BAL is the best specimen. We have tried direct smears and Cyto-spins with much less success. Thanks is advance, Lynne Lynne A. Bell, HT (ASCP) Laboratory Central Vermont Hospital P.O. Box 547 Barre, VT 05641 (802)371-4122 From mohana_g2002 <@t> yahoo.com Tue Jan 10 07:58:45 2006 From: mohana_g2002 <@t> yahoo.com (MOHANA) Date: Tue Jan 10 07:58:55 2006 Subject: [Histonet] cresyl violet Message-ID: <20060110135845.33085.qmail@web33501.mail.mud.yahoo.com> hello !! i am doing a cresyl violet staining to study the neurons in chick hippocampus.....but d staining sometimes does nt cm..... any suggestions why?? n does this stain helps to understaind neuronal morphology ?? mohana Mohana --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less From rjbuesa <@t> yahoo.com Tue Jan 10 09:18:22 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Jan 10 09:18:29 2006 Subject: [Histonet] Stains for pneumocystis In-Reply-To: Message-ID: <20060110151822.106.qmail@web61211.mail.yahoo.com> Hi Lynne: I also used Grocott's Methenamine silver but it took about 25-30 minutes using the following protocol: a- after dewaxing, hydrating → dist. water b- to chromic acid heated x30 secs. in my 480W microwave oven (MWO)[in my MWO this was equivalent to 3.6 kcal of energy; you should adjust the time to the energy output of your MWO in order to not overheat the sections] c- to a water bath at 60?C x 5 min. d-wash with dist water e- to the methenamine working solution → MWO x 45 secs. (= 5.4 kcal energy) f- to 60?C water bath until sections are dark brown (approx. 5-7 min.) g- wash dist. water h- gold chloride sol x 5 min. i- wash with dist water j- sodium thiosulfate sol. x 30 secs. k- wash l- counterstain with light green sol. just a few seconds m-dehydrate, clear, cover slip. This procedure takes less than 30 min. total Hope this will help you. Ren? J.` "Bell, Lynne" wrote: My fellow Histonetters, I would like to know what stain you are using for pneumocystis. We are presently using a "modified" GMS that takes approximately one hour and 15 minutes to perform. A few of our physicians would like something a bit faster. (Of course, I would like to get the specimen sometimes before 3 p.m., but that is another story for another time!) I have also found that a Thin Prep slide from a BAL is the best specimen. We have tried direct smears and Cyto-spins with much less success. Thanks is advance, Lynne Lynne A. Bell, HT (ASCP) Laboratory Central Vermont Hospital P.O. Box 547 Barre, VT 05641 (802)371-4122 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos ? Showcase holiday pictures in hardcover Photo Books. You design it and we?ll bind it! From rjbuesa <@t> yahoo.com Tue Jan 10 09:34:16 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Jan 10 09:34:25 2006 Subject: [Histonet] cresyl violet In-Reply-To: <20060110135845.33085.qmail@web33501.mail.mud.yahoo.com> Message-ID: <20060110153416.74914.qmail@web61215.mail.yahoo.com> Hi Mohana: For neurons I always preferred Tress & Tress formula. Sol.A: dist. water → 100mL + acetic acid→0.01 mL + cresyl violet→ 0.5g Sol.B: chloroform→75mL + abs.ethanol→12.5 mL+ ether→12.5mL Sol.C: hydrochloric acid→0.003 mL in 100 mL 95% ethanol. Sol.D: sodium bicarbonate→ 0.01g + 95% ethanol→ 100 mL Protocol: dewash-hydrate sections→ Sol.Ax 30 min. at 50?C →wash→to% ethanol until no more blue washes out→ sol.B x 2-5 min.→ sol.C until differentiation is complete→ sol.D x 2-3 min→ wash in 95% ethanol→ dehydrate with acetone→clear→cover slip. Hope this will help. Ren? J. MOHANA wrote: hello !! i am doing a cresyl violet staining to study the neurons in chick hippocampus.....but d staining sometimes does nt cm..... any suggestions why?? n does this stain helps to understaind neuronal morphology ?? mohana Mohana --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From Kristopher.Kalleberg <@t> unilever.com Tue Jan 10 09:38:24 2006 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Tue Jan 10 09:38:33 2006 Subject: [Histonet] Abcam Message-ID: Hello, I am purchasing a number of antibodies and it seems that Abcam has a large number of which I need but I have never ordered from them before. Has anyone ordered antibodies from them and what were the results using there products. I just wonder if the results are as good or better as compared to other companies from which I usually order from. I am specifically running IHC for Ki-67, TUNEL, CD1a, thymine dimer, and NFkappa B. Thanks in advance for the help. Kris Kalleberg From DNolan <@t> evanhospital.com Tue Jan 10 10:26:50 2006 From: DNolan <@t> evanhospital.com (Donna M. Nolan) Date: Tue Jan 10 10:30:19 2006 Subject: [Histonet] correction-Leitz microtome Message-ID: <2C7B7A4D7DE62A46BC2EFB70D3ABF9326A49B2@EVANXSCL.evanhospital.net> I'm sorry for the confusion. It is a Leitz microtome not Leica. It was orginally purchased in 1989 and has been in storage since 1997. I do not have a manual for it. I'm not sure of a price. I need to talk to our manager about selling price since people are interested. Donna Nolan Evangelical Community Hospital From tp2 <@t> medicine.wisc.edu Tue Jan 10 10:51:21 2006 From: tp2 <@t> medicine.wisc.edu (Thomas Pier) Date: Tue Jan 10 10:52:17 2006 Subject: [Histonet] Reihard, please help CD4, CD8 in FFPE Bone Marrow Message-ID: Hello, I am looking for Reinhard von Wasielewski. I recently saw a post stating that he had working protocols for CD4 and CD8 that even worked in decalcified bone marrow. CD4 and CD8 in FFPE, decalcified bone marrow is exactly what I'm having problems with. I have been banging my head against the wall with this for some time now. I would really appreciate any help I could get on this. Thank you very much. Tom Pier From edessas <@t> rmy.emory.edu Tue Jan 10 11:01:55 2006 From: edessas <@t> rmy.emory.edu (Evan Dessasau) Date: Tue Jan 10 10:53:40 2006 Subject: [Histonet] Alkaline Congo Red Method (Puchtler et al) Message-ID: <43C3E883.5040006@rmy.emory.edu> Hi.....being new to histology as a carrere I have trouble getting stains to look like they do in the book. The current issue I have is with the Puchtler's Alkaline Congo Red. The nuclear staining is done before the alkaline solutions. At the end of the protocol the nuclei are no longer blue. You can almost watch the color change occur in the alkaline salt solution. The protocol calls for 2.5 min. in the Harris hematoxylin and then rinse for several min. I've not found that the nuclei come to a good blue color this way and the color fades. When I follow the staining time on the bottle for the Harris, which calls for 10min. and then rinse for more like 14 to 21 min. I get very good blue color. I can still watch this fade to a dull color which is unacceptable. I'm using Harris from EMS. I do filter and I follow the protocol in the text Histotechnology: A self-instructional Text by Carson. What am I not doing right ?? Thank you for what ever advice , E-van From laurie.colbert <@t> huntingtonhospital.com Tue Jan 10 11:18:26 2006 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Tue Jan 10 11:18:42 2006 Subject: [Histonet] Stains for pneumocystis Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628005653934@EXCHANGE1.huntingtonhospital.com> Lynne, We run a Grocott's Methenamine Silver, utilizing the microwave. We stain exactly as the protocol states, except we microwave on the silver step (we have a regular household microwave). Make up the silver as directed, put in a plastic 5-slide holder, and microwave on high for 30-60 seconds. Just keep on eye on the silver so it doesn't boil over. I check my slide at about 20 seconds and then every 10 seconds after that. When my control section is a golden brown color, I take the slide out, rinse in DI water, and proceed with staining. We can complete the stain in less than 20 minutes. Laurie Colbert Huntington Hospital -----Original Message----- From: Bell, Lynne [mailto:Lynne.Bell@hitchcock.org] Sent: Tuesday, January 10, 2006 5:22 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Stains for pneumocystis My fellow Histonetters, I would like to know what stain you are using for pneumocystis. We are presently using a "modified" GMS that takes approximately one hour and 15 minutes to perform. A few of our physicians would like something a bit faster. (Of course, I would like to get the specimen sometimes before 3 p.m., but that is another story for another time!) I have also found that a Thin Prep slide from a BAL is the best specimen. We have tried direct smears and Cyto-spins with much less success. Thanks is advance, Lynne Lynne A. Bell, HT (ASCP) Laboratory Central Vermont Hospital P.O. Box 547 Barre, VT 05641 (802)371-4122 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kaleid11 <@t> yahoo.com Tue Jan 10 11:21:35 2006 From: kaleid11 <@t> yahoo.com (Adam Perry) Date: Tue Jan 10 11:21:49 2006 Subject: [Histonet] Abcam In-Reply-To: Message-ID: <20060110172135.31830.qmail@web30409.mail.mud.yahoo.com> I've ordered from Abcam before and didn't have a problem with the antibodies...I'm not sure if Abcam actually makes all its antibodies in-house though. For instance, the antibody I purchased is actually made by one company (Lifespan Bioscience) and then distributed through multiple companies (Abcam, Novus Biologicals, etc.)...So you may want to check the specifics on each antibody (i.e. track down where they are actually made) and ask people about that source... Hope that helps, Adam Adam Perry Department of Physiology and Biophysics University of Illinois Chicago, IL 60612 Kristopher Kalleberg wrote: Hello, I am purchasing a number of antibodies and it seems that Abcam has a large number of which I need but I have never ordered from them before. Has anyone ordered antibodies from them and what were the results using there products. I just wonder if the results are as good or better as compared to other companies from which I usually order from. I am specifically running IHC for Ki-67, TUNEL, CD1a, thymine dimer, and NFkappa B. Thanks in advance for the help. Kris Kalleberg _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From gcallis <@t> montana.edu Tue Jan 10 11:38:01 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Jan 10 11:38:26 2006 Subject: [Histonet] Stains for pneumocystis In-Reply-To: References: Message-ID: <6.0.0.22.1.20060110102729.01b3caf8@gemini.msu.montana.edu> We have one lab whose expertise is Pneuocystis research. They use Dade-Behring Diff-Quick from VWR but extend time in Solution 2. Protocol: Fix cells with methanol Solution 1 - according to package insert Solution 2 - 30 to 40 minutes Rinse per package insert, air dry and look under oil, a coverslip can be mounted over an air dry slide (fans help out with drying or forced air of some sort). Any blood cells will be too dark but the Pneumocystis needs the extra time in Sol. 2 in order to stain the cytoplasm for viewing. This is basically a Wright Giemsa stain, and if done like a blood smear, should probably work also. Wright Giemsa is available under Harleco brand. One might have to stain longer after buffering however. They stain too numerous to count murine BAL samples, spun down with Cytospin. GMS can be shortened by using a MW staining method and you may want to fix the smears/spins differently when using this protocol i.e. other than methanol. Good luck At 06:22 AM 1/10/2006, you wrote: >My fellow Histonetters, > >I would like to know what stain you are using for pneumocystis. We are >presently using a "modified" GMS that takes approximately one hour and >15 minutes to perform. A few of our physicians would like something a >bit faster. (Of course, I would like to get the specimen sometimes >before 3 p.m., but that is another story for another time!) I have also >found that a Thin Prep slide from a BAL is the best specimen. We have >tried direct smears and Cyto-spins with much less success. > >Thanks is advance, >Lynne > >Lynne A. Bell, HT (ASCP) >Laboratory >Central Vermont Hospital >P.O. Box 547 >Barre, VT 05641 >(802)371-4122 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gentras <@t> vetmed.auburn.edu Tue Jan 10 11:38:29 2006 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Tue Jan 10 11:38:56 2006 Subject: Fwd: Re: [Histonet] cresyl violet Message-ID: <6.0.1.1.0.20060110113827.01b41ca0@mailhost.vetmed.auburn.edu> >Date: Tue, 10 Jan 2006 11:37:51 -0600 >To: MOHANA >From: "Atoska S. Gentry" >Subject: Re: [Histonet] cresyl violet > > >Hello Mohana you might try using cresyl violet in conjunction with luxol >fast blue (LFB) as in Kluver - Barrera Method for Myelin and Nerve Cells. >The protocol I use for this stain comes from the 3rd edition of the >AFIP Manual of Histologic Staining Methods pp. 203-204. Best wishes. Atoska > >At 07:58 AM 1/10/2006, you wrote: >>hello !! >> >> i am doing a cresyl violet staining to study the neurons in chick >> hippocampus.....but d staining sometimes does nt cm..... any suggestions why?? >> n does this stain helps to understaind neuronal morphology ?? >> >> mohana >> >> >> >>Mohana >> >> >>--------------------------------- >> Yahoo! DSL Something to write home about. Just $16.99/mo. or less >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MElliott <@t> mrl.ubc.ca Tue Jan 10 11:51:02 2006 From: MElliott <@t> mrl.ubc.ca (Mark Elliott) Date: Tue Jan 10 11:51:49 2006 Subject: [Histonet] Rabbit IL-6 Message-ID: <43C38386020000D600002563@mail.mrl.ubc.ca> Does anyone know of an antibody (monoclonal or polyclonal) against Rabbit IL-6? If not, what companies offer custom antibody production, in addition to Invitrogen? thanks Mark Elliott Vancouver, BC From Jan.Minshew <@t> leica-microsystems.com Tue Jan 10 11:54:57 2006 From: Jan.Minshew <@t> leica-microsystems.com (Jan.Minshew@leica-microsystems.com) Date: Tue Jan 10 11:55:12 2006 Subject: [Histonet] correction-Leitz microtome In-Reply-To: <2C7B7A4D7DE62A46BC2EFB70D3ABF9326A49B2@EVANXSCL.evanhospital.net> Message-ID: Hi Donna, You were actually correct. The Leitz 1512 is part of the Leica family. It's one of our Grandaddy's...like the AO820. We have both models in our microtome museum here is the US (not that they're that old--I used them daily). There are some REALLY old microtomes in our facility in Nussloch, Germany. Best wishes to all for a great 2006, Jan Minshew HT(ASCP)HTL Marketing Manager Leica Microsystems, Inc. 2345 Waukegan Rd. Bannockburn, IL 60015 800.248.0123 x7015 (toll free) 847.405.7051 (direct) 847.405.7941 (fax) "Donna M. Nolan" To Sent by: "Histonet" histonet-bounces@ lists.utsouthwest cc ern.edu Subject [Histonet] correction-Leitz 01/10/2006 10:26 microtome AM I'm sorry for the confusion. It is a Leitz microtome not Leica. It was orginally purchased in 1989 and has been in storage since 1997. I do not have a manual for it. I'm not sure of a price. I need to talk to our manager about selling price since people are interested. Donna Nolan Evangelical Community Hospital _______________________________________________ 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 gcallis <@t> montana.edu Tue Jan 10 12:01:29 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Jan 10 12:01:41 2006 Subject: [Histonet] Abcam In-Reply-To: <20060110172135.31830.qmail@web30409.mail.mud.yahoo.com> References: <20060110172135.31830.qmail@web30409.mail.mud.yahoo.com> Message-ID: <6.0.0.22.1.20060110105953.01b71710@gemini.msu.montana.edu> Adam gave some good advice and also include what species are you staining, mouse, rat, in your question. >.So you may want to check the specifics on each antibody (i.e. track down >where they are actually made) and ask people about that source... > > Hope that helps, > Adam > > Adam Perry > Department of Physiology and Biophysics > University of Illinois > Chicago, IL 60612 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Tue Jan 10 12:06:47 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Jan 10 12:07:07 2006 Subject: [Histonet] microwave GMS for pneumocystis and possible boil over containment In-Reply-To: <0BE6ADFAE4E7E04496BF21ABD346628005653934@EXCHANGE1.hunting tonhospital.com> References: <0BE6ADFAE4E7E04496BF21ABD346628005653934@EXCHANGE1.huntingtonhospital.com> Message-ID: <6.0.0.22.1.20060110110213.01b82e88@gemini.msu.montana.edu> Laurie, Very nice and quick GMS here!! To prevent boil over spills in MW chamber, place slide holder upright in a zip lock baggie, with one corner left open for venting. This very tidy hint came from Cheryl Crowder, a MW staining expert. At 10:18 AM 1/10/2006, you wrote: >Lynne, > >We run a Grocott's Methenamine Silver, utilizing the microwave. We stain >exactly as the protocol states, except we microwave on the silver step (we >have a regular household microwave). >Make up the silver as directed, put in a plastic 5-slide holder, and >microwave on high for 30-60 seconds. Just keep on eye on the silver so it >doesn't boil over. I check my slide at about 20 seconds and then every 10 >seconds after that. When my control section is a golden brown color, I >take the slide out, rinse in DI water, and proceed with staining. We can >complete the stain in less than 20 minutes. > >Laurie Colbert >Huntington Hospital > >-----Original Message----- >From: Bell, Lynne [mailto:Lynne.Bell@hitchcock.org] >Sent: Tuesday, January 10, 2006 5:22 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Stains for pneumocystis > > >My fellow Histonetters, > >I would like to know what stain you are using for pneumocystis. We are >presently using a "modified" GMS that takes approximately one hour and >15 minutes to perform. A few of our physicians would like something a >bit faster. (Of course, I would like to get the specimen sometimes >before 3 p.m., but that is another story for another time!) I have also >found that a Thin Prep slide from a BAL is the best specimen. We have >tried direct smears and Cyto-spins with much less success. > >Thanks is advance, >Lynne > >Lynne A. Bell, HT (ASCP) >Laboratory >Central Vermont Hospital >P.O. Box 547 >Barre, VT 05641 >(802)371-4122 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Tue Jan 10 12:18:54 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Jan 10 12:19:11 2006 Subject: [Histonet] Rabbit IL-6 In-Reply-To: <43C38386020000D600002563@mail.mrl.ubc.ca> References: <43C38386020000D600002563@mail.mrl.ubc.ca> Message-ID: <6.0.0.22.1.20060110111742.01ba6b98@gemini.msu.montana.edu> Have your tried R&D Systems, they are cytokine antibody gurus, and may be able to help you find what you need if they do not have it. At 10:51 AM 1/10/2006, you wrote: >Does anyone know of an antibody (monoclonal or polyclonal) against Rabbit >IL-6? If not, what companies offer custom antibody production, in addition >to Invitrogen? Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From pruegg <@t> ihctech.net Tue Jan 10 13:16:06 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Tue Jan 10 13:16:07 2006 Subject: [Histonet] Stem Cell Factor Message-ID: <200601101915.k0AJFuwZ000458@chip.viawest.net> I need advise for doing Stem Cell Factor (SCF) IHC on ffpe human skin tissue. What is a good antibody for this? Protocol? Thank you, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From portera <@t> msu.edu Tue Jan 10 13:20:44 2006 From: portera <@t> msu.edu (Amy Porter) Date: Tue Jan 10 13:21:53 2006 Subject: [Histonet] Slide Press? Message-ID: <003801c6161a$f3cf07b0$8e7a0923@HistoJJ> I have had a question posed to me by our e.m. technician about a "slide press" to weigh down section of bone that have been embedded in resin. Does anyone out there know what this is or know of a vendor where one may be purchased. Thanks in advance for all of the responses. Amy S. Porter, HT(ASCP) QIHC - Supervisor Michigan State University Laboratory for Investigative HistoPathology Department of Physiology / Division of Human Pathology 2100 Biomedical Physical Sciences Bldg. Room #2133 East Lansing, MI 48824-3320 Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 Email: portera@msu.edu www.humanpathology.msu.edu From RebeNoel <@t> aol.com Tue Jan 10 13:46:26 2006 From: RebeNoel <@t> aol.com (RebeNoel@aol.com) Date: Tue Jan 10 13:46:44 2006 Subject: [Histonet] Histology Positions Message-ID: <23d.500cdfe.30f56912@aol.com> Dear Histology Professionals: I am a recruiter that specializes in the placement of Histology Professionals and other laboratory disciplines. I work with many companies throughout the united states that are in search of all levels of Histology professionals. I am looking for people who are searching for a new position. I am very familiar with the histology arena and understand it is often difficult to seek new employment because everyone knows everyone in the histology world. That's why working with a recruiter will enable you to maintain confidentiality well conducting your job search. If you are interested in finding out about open positions please email me your resume or contact info. I can be reached at: rebenoel@aol.com Thank you, Rebecca Noel Executive Match Personnel. 919-601-1946 From mlb <@t> nmr.mgh.harvard.edu Tue Jan 10 13:55:12 2006 From: mlb <@t> nmr.mgh.harvard.edu (mlb@nmr.mgh.harvard.edu) Date: Tue Jan 10 13:55:21 2006 Subject: [Histonet] "MBS" in Luxol Fast Blue MBS Message-ID: <2022.132.183.203.6.1136922912.squirrel@mail.nmr.mgh.harvard.edu> Hi, I've been using luxol fast blue MBS for myelin staining and would like to know what the acronym "MBS" represents. I have read many primary sources, such as articles by Kluver and Barrera as well as Salthouse, and I have performed internet searches with Google, all to no avail (other than finding histonet!). Thanks for your help! Megan =-) p.s. Also, any explanations of the other acronyms used in luxol dyes (ARN, G and the "N" in MBSN) would be much appreciated! From gcallis <@t> montana.edu Tue Jan 10 13:55:42 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Jan 10 13:55:58 2006 Subject: [Histonet] Slide Press? In-Reply-To: <003801c6161a$f3cf07b0$8e7a0923@HistoJJ> References: <003801c6161a$f3cf07b0$8e7a0923@HistoJJ> Message-ID: <6.0.0.22.1.20060110124616.01b437f8@gemini.msu.montana.edu> We placed are slides between sheets of polyethylene (cut from a thicker baggie) with a sheet between each slide, then clamped the slides between pieces of wood so as to NOT break the glass slides, using C clamps from a hardward store to apply pressure evenly. Clamps come in all sizes, but one can find them at Walmart too. You can be as creative on this as you want - something cheap, fast and efficient. At 12:20 PM 1/10/2006, you wrote: >I have had a question posed to me by our e.m. technician about a "slide >press" to weigh down section of bone that have been embedded in >resin. Does anyone out there know what this is or know of a vendor where >one may be purchased. Thanks in advance for all of the responses. > >Amy S. Porter, HT(ASCP) QIHC - Supervisor >Michigan State University >Laboratory for Investigative HistoPathology >Department of Physiology / Division of Human Pathology >2100 Biomedical Physical Sciences Bldg. Room #2133 >East Lansing, MI 48824-3320 >Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 >Email: portera@msu.edu >www.humanpathology.msu.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From dellav <@t> musc.edu Tue Jan 10 14:32:38 2006 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Tue Jan 10 14:33:19 2006 Subject: [Histonet] Have you ever considered teaching ? Message-ID: If you've not considered teaching, I would encourage you to do so. few things are more rewarding than stimulating and witnessing the enthusiasm and awe on the faces of young people who are embarking on careers in histotechnology. In 2004, I started a Histotechnology School (HTL) at my facility because I wanted to "pay it forward" in repayment, so to speak, for the assistance and encouragment I received from others when I was a fledgling histotech. This is a new venture for me, and we are mid year in our second class. I will tell you that the joy that I have working with the students is greater than I could have imagined. so, if you think that teaching might interest you, and you meet our posting requirements, feel free to visit us at www.musc.edu/histoprogram . You may formally apply for position #A0516067 at https://www.applymuscjobs.com/applicants our position is posted at the NSH website and will appear in Advance for Medical Laboratory Professionals. While I am looking for an individual with prior teaching experience, this can be in many forms including conducting workshops at conferences or providing instruction in other settings. However, prior teaching experience is not an absolute requirement and I will consider individuals with strong clinical skills. our posting reads as follows: Come join the excitement at the MUSC Medical Center in Charleston, South Carolina. Explore employment opportunities in our state of the art facility as you explore our city. Visitors travel to Charleston each year to walk down the cobblestone streets, relive its history and enjoy the beaches. Experienced instructor wanted for newly accredited, 12 month, post-baccalaureate certificate Histotechnology Program (HTL). Details about our program may be found at www.musc.edu/histoprogram The successful candidate will possess a bachelor of science degree in biology, medical technology or other life science and at least five years of clinical histology experience with a strong emphasis in special stains and immunohistochemistry. A Master's degree in science, education or healthcare administration or equivalent is preferred. Certification in histotechnology, with the American Society of Clinical Pathologists (ASCP) at the technologist level (HTL) is required. Certification at the technician level may be substituted with ten or more years of clinical histology experience that include emphasis in the areas of specialization indicated above. Prior teaching experience is strongly preferred but not required. Applicants will be required to conduct a formal presentation to the selection committee at interview. Interested applicants are invited to contact Vinnie Della Speranza at dellav@musc.edu From Barry.R.Rittman <@t> uth.tmc.edu Tue Jan 10 15:17:12 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Tue Jan 10 15:17:16 2006 Subject: [Histonet] Slide Press? Message-ID: Amy We had a similar problem with ground sections and also sheets of epithelium from mouse ears. Our solution was to use weights made by cutting solid brass cylinders (3/4" diameter and 2" long) and placing these weights over the area under the cover glass where section was located. This worked very well for us even if sections were slightly wedge shaped. This also allows you to add additional mountant if air tends to creep in. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Tuesday, January 10, 2006 1:56 PM To: Amy Porter; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Slide Press? We placed are slides between sheets of polyethylene (cut from a thicker baggie) with a sheet between each slide, then clamped the slides between pieces of wood so as to NOT break the glass slides, using C clamps from a hardward store to apply pressure evenly. Clamps come in all sizes, but one can find them at Walmart too. You can be as creative on this as you want - something cheap, fast and efficient. At 12:20 PM 1/10/2006, you wrote: >I have had a question posed to me by our e.m. technician about a "slide >press" to weigh down section of bone that have been embedded in >resin. Does anyone out there know what this is or know of a vendor where >one may be purchased. Thanks in advance for all of the responses. > >Amy S. Porter, HT(ASCP) QIHC - Supervisor >Michigan State University >Laboratory for Investigative HistoPathology >Department of Physiology / Division of Human Pathology >2100 Biomedical Physical Sciences Bldg. Room #2133 >East Lansing, MI 48824-3320 >Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 >Email: portera@msu.edu >www.humanpathology.msu.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pathrm35 <@t> adelphia.net Tue Jan 10 16:23:14 2006 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Tue Jan 10 16:23:26 2006 Subject: [Histonet] histologists position in Southeast FL Message-ID: <8463034.1136931794767.JavaMail.root@web15> Fellow Tech's, We are seeking a histology technologist for our new dermpath lab in Southeast FL. This is a M-F, 9AM-5:30 PM position with no weekends or nights. Duties include IHC, grossing, embedding and sectioning. Requirements are: HT or HTL (ASCP), IHC experience a plus, Florida licensed or eligible Histology Technologist and able to gross tissue under CLIA regs (AS or BS degree in a science field). As per my administrator: relocation/sign on bonus for qualified candidates and salary range of $25-$28/hour. Contact: Ron Martin fax 561-721-1249 From vazquezr <@t> ohsu.edu Tue Jan 10 16:48:10 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Tue Jan 10 16:49:19 2006 Subject: [Histonet] positive rat bladder control Message-ID: Hello, I have been getting positive rat bladder control solution for paraneoplastic/pemphigus from Immco and now they are not stocking it. Would anyone know of another source? Thanks in advance, Robyn OHSU From lrichey <@t> u.washington.edu Tue Jan 10 17:02:33 2006 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Tue Jan 10 17:02:46 2006 Subject: [Histonet] Reihard, please help CD4, CD8 in FFPE Bone Marrow In-Reply-To: References: Message-ID: <43C43D09.1010605@u.washington.edu> We use both CD4 and CD8 on FFPE sections. We had problems with the CD4 until we changed our quenching step to 0.5% H2O2 in MEOH for 10 min. The pretreatment for both is 15 min microwave in pH 8 EDTA. Thomas Pier wrote: >Hello, >I am looking for Reinhard von Wasielewski. I recently saw a post >stating that he had working protocols for CD4 and CD8 that even worked >in decalcified bone marrow. CD4 and CD8 in FFPE, decalcified bone >marrow is exactly what I'm having problems with. I have been banging my >head against the wall with this for some time now. I would really >appreciate any help I could get on this. Thank you very much. > >Tom Pier > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From shive003 <@t> umn.edu Tue Jan 10 17:25:13 2006 From: shive003 <@t> umn.edu (Jan Shivers) Date: Tue Jan 10 17:25:26 2006 Subject: [Histonet] Feline Calicivirus IHC Message-ID: <00a601c6163d$1bceed80$41065486@auxs.umn.edu> Does anyone have some helpful hints as to how to get consistent staining with FCV IHC? Or is this one of those buggers (sorry, to those in the UK) which require positive controls to be sectioned as needed, and not stored for periods of time? Currently I'm trying two different monoclonals, with enzyme tissue pretreatment, and EnVision+/HRP polymer detection system. Sometimes I can get a handful of cells to stain in the infected lungs; sometimes they like to hide from me. Much appreciation for any comments you may have from your experiences. Jan Shivers U of MN Vet Diag Lab From ewrona <@t> yahoo.com Tue Jan 10 18:52:22 2006 From: ewrona <@t> yahoo.com (Erin Wrona) Date: Tue Jan 10 18:52:31 2006 Subject: [Histonet] carbowax Message-ID: <20060111005222.12149.qmail@web52505.mail.yahoo.com> Hi all, Our lab director wants us to try using carbowax polyethylene glycol as an embedding medium because it has a slightly higher melting point than our paraffin. I have never used it other than in formulations for frozen sections, in which case it is water soluble. One of the other techs here thinks that it is always water soluble. Does anyone know for sure? The specifics are: ems product #19770, PEG 8000 flakes, melting point 60-63 degrees. Thanks in advance, Erin Wrona San Francisco, CA From bhewlett <@t> cogeco.ca Tue Jan 10 19:25:38 2006 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Tue Jan 10 19:25:48 2006 Subject: [Histonet] carbowax References: <20060111005222.12149.qmail@web52505.mail.yahoo.com> Message-ID: <000301c6164d$edc632c0$6400a8c0@mainbox> Erin, Why does the lab director think you need a higher melting point wax? If a higher melting point is really necessary, buy a paraffin wax with one. Substitution of Carbowax for this purpose only, will lead to nothing but endless frustration and heartbreak for the techs! Yes, it is water soluble!!! Blocks will have to be stored desiccated or coated with paraffin wax. Sectioning in conditions of high humidity can be a real challenge. Floating out can be a total beast, don't use water since the violent diffusion currents will disrupt the sections. Addition of soap or up to 20% PEG 900 can minimize this, but trust me it's still a beast! Carbowax has a few special uses, it avoids the use of dehydration and clearing solvents and results in less shrinkage. It is definitely not intended for routine use. Bryan ----- Original Message ----- From: "Erin Wrona" To: Sent: Tuesday, January 10, 2006 7:52 PM Subject: [Histonet] carbowax > Hi all, > > Our lab director wants us to try using carbowax polyethylene glycol as an > embedding medium because it has a slightly higher melting point than our > paraffin. I have never used it other than in formulations for frozen > sections, in which case it is water soluble. One of the other techs here > thinks that it is always water soluble. Does anyone know for sure? > > The specifics are: ems product #19770, PEG 8000 flakes, melting point > 60-63 degrees. > > Thanks in advance, > Erin Wrona > San Francisco, CA > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From christy0424 <@t> gmail.com Tue Jan 10 19:55:50 2006 From: christy0424 <@t> gmail.com (Kuo Christy) Date: Tue Jan 10 19:56:00 2006 Subject: [Histonet] Re: Reihard, please help CD4, CD8 in FFPE Bone Marrow In-Reply-To: <43C43D09.1010605@u.washington.edu> References: <43C43D09.1010605@u.washington.edu> Message-ID: <5e4834130601101755n5109e9c6l133e9f1ec44d31c6@mail.gmail.com> I've treated the H2O2 step. However, my background is very high. I was wondering if you treated any other step and your antibody incubation time and concentration. Thank you in advance for answering~ sincerely, Christy On 1/11/06, Lori Richey wrote: > We use both CD4 and CD8 on FFPE sections. We had problems with the CD4 > until we changed our quenching step to 0.5% H2O2 in MEOH for 10 min. The > pretreatment for both is 15 min microwave in pH 8 EDTA. > > Thomas Pier wrote: > > >Hello, > >I am looking for Reinhard von Wasielewski. I recently saw a post > >stating that he had working protocols for CD4 and CD8 that even worked > >in decalcified bone marrow. CD4 and CD8 in FFPE, decalcified bone > >marrow is exactly what I'm having problems with. I have been banging my > >head against the wall with this for some time now. I would really > >appreciate any help I could get on this. Thank you very much. > > > >Tom Pier > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From louise.renton <@t> gmail.com Wed Jan 11 00:47:43 2006 From: louise.renton <@t> gmail.com (louise renton) Date: Wed Jan 11 00:47:55 2006 Subject: [Histonet] Slide Press? In-Reply-To: <003801c6161a$f3cf07b0$8e7a0923@HistoJJ> References: <003801c6161a$f3cf07b0$8e7a0923@HistoJJ> Message-ID: We use a PROXXON model maker's vice. this allows one to clamp slides and transport them to the incubator. If you want, I can look up the model number for you, as I do not have it to hand right now best regards On 1/10/06, Amy Porter wrote: > I have had a question posed to me by our e.m. technician about a "slide press" to weigh down section of bone that have been embedded in resin. Does anyone out there know what this is or know of a vendor where one may be purchased. Thanks in advance for all of the responses. > > Amy S. Porter, HT(ASCP) QIHC - Supervisor > Michigan State University > Laboratory for Investigative HistoPathology > Department of Physiology / Division of Human Pathology > 2100 Biomedical Physical Sciences Bldg. Room #2133 > East Lansing, MI 48824-3320 > Phone: (517) 355-6475 ext 1480 / Fax: (517) 432-1368 > Email: portera@msu.edu > www.humanpathology.msu.edu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....onwards through the fog!" From wasielewski.reinhard.von <@t> mh-hannover.de Wed Jan 11 01:38:03 2006 From: wasielewski.reinhard.von <@t> mh-hannover.de (wasielewski.reinhard.von@mh-hannover.de) Date: Wed Jan 11 01:38:14 2006 Subject: [Histonet] CD4 and CD8 in human FFPE In-Reply-To: Message-ID: <43C4C3EB.16576.B3807CF9@localhost> Dear Histonetters, I've got many requests asking for our protocol for staining CD4 and CD8, - therefore I think it is best to answer in general on the histonet for all users. Since I was out- house for several days, I apologize for any delay in my answer ! We make pretreatment for CD8 and CD4 in a household microwave oven, place the slides well submerged in buffer (0,8 liter container) for 30 minutes, using 800 Watt. For bone-marrow (decalified by EDTA in a ultrasonic bath), we use as buffer monocitrate pH 6.0. For all regular human tissues, we use monocitric buffer pH 7.2 For CD4, we use Zymed "ready-to-use" monoclonal antibody 08-0101. For CD8, we use Dako antibody (M7103) in a dilution 1:40 For both markers, detection is done with the Zymed detection kit "Zytochem plus HRP 5000". All stainings are done in daily routine on a Lab-Vision Autostainer. Sorry, but we have only experience in human FFPE tissue. I have no idea if it works in bovine tissue as well. In mouse tissue, it doesn't work - that's why I posted my initial request to histonet to get protocols for mouse FFPE. All the answers I got hint to the fact that there are no reliable protocols or antibodies for CD4/8 in mouse FFPE so far. However, we will not give up and try harder to solve this dilemma - as soon as we have answers, I'll let you know ! Best regards - and thanks for the many answers and comments from you. Have a successful new years 2006, - with less war on earth and more thruth. Sincerely yours Reinhard. Date sent: Mon, 19 Dec 2005 19:10:42 +0100 From: histonet-request@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 25, Issue 24 To: histonet@lists.utsouthwestern.edu Send reply to: histonet@lists.utsouthwestern.edu [ Double-click this line for list subscription options ] 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. CD4 and CD8 in mouse / human FFPE (wasielewski.reinhard.von@mh-hannover.de) 2. Re: CD4 and CD8 in mouse / human FFPE (Laurie Reilly) ---------------------------------------------------------------------- Message: 1 Date: Sun, 18 Dec 2005 20:49:21 +0100 From: wasielewski.reinhard.von@mh-hannover.de Subject: [Histonet] CD4 and CD8 in mouse / human FFPE To: histonet@lists.utsouthwestern.edu Message-ID: <43A5CB51.20138.3A8527E7@localhost> Content-Type: text/plain; charset=US-ASCII Hi Patsy, we do a lot of CD4 and CD8 in human FFPE tissues, even in BM after decalification. It works very good and is daily routine in our lab. If you need advice, give me a mail. We now got some hints how both markers could work in mouse tissue, too. If we succeed, we'll let you know. best regards Reinhard. PD Dr. med. Reinhard von Wasielewski ------------------------------ Message: 2 Date: Mon, 19 Dec 2005 09:09:49 +1000 From: Laurie Reilly Subject: Re: [Histonet] CD4 and CD8 in mouse / human FFPE To: wasielewski.reinhard.von@mh-hannover.de Cc: histonet Message-ID: <5.1.0.14.0.20051219090417.00c4a238@mail.jcu.edu.au> Content-Type: text/plain; charset="us-ascii"; format=flowed Reinhard, Could I please have a copy of your protocol, we have had lots of trouble trying to stain with CD4 and CD8 in bovine tissues. Any advice you can give would be most welcome, particularly about antigen retrieval. Hope everyone has a happy Christmas, Regards, Laurie. At 08:49 PM 18/12/2005 +0100, you wrote: >Hi Patsy, >we do a lot of CD4 and CD8 in human FFPE tissues, even in BM after >decalification. >It works very good and is daily routine in our lab. If you need advice, >give me a mail. >We now got some hints how both markers could work in mouse tissue, too. If we >succeed, we'll let you know. > >best regards >Reinhard. > > > >PD Dr. med. Reinhard von Wasielewski > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 25, Issue 24 **************************************** PD Dr. med. Reinhard von Wasielewski From c.m.vanderloos <@t> amc.uva.nl Wed Jan 11 01:50:37 2006 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Wed Jan 11 01:50:48 2006 Subject: [Histonet] RE: Ki-67 or PCNA on FFPE primate tissue Message-ID: <3fc8b33fc76a.3fc76a3fc8b3@amc.uva.nl> Roger, In my hands the rabbit monoclonal anti-Ki67 antibody, clone SP6 (LabVision) stained beautifully human, mouse and rat tissues. I think you have a very good chance with this antibody for primate tissues. Good luck! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Mon, 9 Jan 2006 11:10:33 -0800 (PST) From: Roger Moretz Subject: [Histonet] Ki-67 or PCNA on FFPE primate tissue To: Histonet@lists.utsouthwestern.edu Anyone with experience using any of the available antibodies for FFPE primate tissue? Any preferences? Roger Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals,Inc. Ridgefield, CT From antje.marcantonio <@t> novartis.com Wed Jan 11 02:02:15 2006 From: antje.marcantonio <@t> novartis.com (antje.marcantonio@novartis.com) Date: Wed Jan 11 02:02:27 2006 Subject: [Histonet] Ki-67 or PCNA on FFPE primate tissue Message-ID: Roger, we do successfully IHC for Ki67 on primate FFPE tissue with the mouse monoclonal anti-Ki67, clone MIB5 from DakoCytomation. Antigen retrieval 98 degrees C with citrate buffer, pH 6.0 for 20 min Primary ab dilution 1/100 for 1 hour , or 1/500 for over night incubation. Hope this helps, Antje Marcantonio Novartis Pharma AG PH214217, AT TX: LAB. WIECZOREK CHBS, WSJ-386.5.55 Novartis Pharma AG Lichtstrasse 35 CH-4056 Basel Switzerland Phone: +41 61 3246730 Fax: +41 61 3247534 Email : antje.marcantonio@novartis.com From rjbuesa <@t> yahoo.com Wed Jan 11 07:53:50 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Jan 11 07:53:58 2006 Subject: [Histonet] carbowax In-Reply-To: <20060111005222.12149.qmail@web52505.mail.yahoo.com> Message-ID: <20060111135350.62591.qmail@web61220.mail.yahoo.com> Hi Erin: Many years ago Carbowax was considered "a blessing" because you could infiltrate tissue (after fixation) with it avoiding dehydration and clearing. This sounds great but it was a mess to store and cut because of this same inherent quality of being water soluble (or miscible,as you wish). If you need harder paraffin (to cut hard tissues) you just simply should try to buy high melting point paraffin. When I worked with plant stems I used to infiltrate them with a heigh melting point paraffin (63-65?C) manufactured on those days by Merck (Darmstadt). I am sure that high melting point paraffins can be found in the market. I thing you should avoid carbowax. Hope this will help. Ren? J. Erin Wrona wrote: Hi all, Our lab director wants us to try using carbowax polyethylene glycol as an embedding medium because it has a slightly higher melting point than our paraffin. I have never used it other than in formulations for frozen sections, in which case it is water soluble. One of the other techs here thinks that it is always water soluble. Does anyone know for sure? The specifics are: ems product #19770, PEG 8000 flakes, melting point 60-63 degrees. Thanks in advance, Erin Wrona San Francisco, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos ? Showcase holiday pictures in hardcover Photo Books. You design it and we?ll bind it! From Kristopher.Kalleberg <@t> unilever.com Wed Jan 11 08:16:13 2006 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Wed Jan 11 08:16:27 2006 Subject: [Histonet] improper fixation Message-ID: All, Last week I was unable to collect 18 biopsies 2mm biopsies for a clinical that is being run and had a colleague collect them for me. After running routine H&E and Fontana Mason stains I found that two biopsies were improperly fixed. Since I have never had this problem before, I was wondering if it is possible to run the sample backwards through paraffin, xylenes, and alcohols and then fix the samples again in 10% NBF and reprocess to paraffin. If this is possible doe sanyone have a protocol or know what the effects will be by doing this? Thank you in advance. Kris Kalleberg From Barry.R.Rittman <@t> uth.tmc.edu Wed Jan 11 08:36:35 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Wed Jan 11 08:36:39 2006 Subject: [Histonet] improper fixation Message-ID: Kris While you can do this, the damage has been done and it is unlikely that you will achieve a better fixation. Also in going back through xylenes and alcohols you will probably cause greater mechanical damage. Even though you processed the tissues, a certain amount of fixation or post fixation has occurred in the alcohols used in processing. Might I suggest that you could try to mordant the sections after they have been deparaffinized and bring down to water? Can use Bouin's or a variety of other solutions that may improve the staining for you. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristopher Kalleberg Sent: Wednesday, January 11, 2006 8:16 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] improper fixation All, Last week I was unable to collect 18 biopsies 2mm biopsies for a clinical that is being run and had a colleague collect them for me. After running routine H&E and Fontana Mason stains I found that two biopsies were improperly fixed. Since I have never had this problem before, I was wondering if it is possible to run the sample backwards through paraffin, xylenes, and alcohols and then fix the samples again in 10% NBF and reprocess to paraffin. If this is possible doe sanyone have a protocol or know what the effects will be by doing this? Thank you in advance. Kris Kalleberg _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Wed Jan 11 08:43:15 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Wed Jan 11 08:43:22 2006 Subject: [Histonet] carbowax Message-ID: Erin I agree totally with Rene. Carbowax is a real pain to work with. To harden the wax you have you may want to try adding Ceresin, a higher MP wax. If you need a much harden wax than this then the ester waxes are more suitable although they also have their problems. A pertinent question here is why specifically does your lab director want or need a higher MP wax, what problems are you having and what tissues are you processing? Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Wednesday, January 11, 2006 7:54 AM To: Erin Wrona; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] carbowax Hi Erin: Many years ago Carbowax was considered "a blessing" because you could infiltrate tissue (after fixation) with it avoiding dehydration and clearing. This sounds great but it was a mess to store and cut because of this same inherent quality of being water soluble (or miscible,as you wish). If you need harder paraffin (to cut hard tissues) you just simply should try to buy high melting point paraffin. When I worked with plant stems I used to infiltrate them with a heigh melting point paraffin (63-65?C) manufactured on those days by Merck (Darmstadt). I am sure that high melting point paraffins can be found in the market. I thing you should avoid carbowax. Hope this will help. Ren? J. Erin Wrona wrote: Hi all, Our lab director wants us to try using carbowax polyethylene glycol as an embedding medium because it has a slightly higher melting point than our paraffin. I have never used it other than in formulations for frozen sections, in which case it is water soluble. One of the other techs here thinks that it is always water soluble. Does anyone know for sure? The specifics are: ems product #19770, PEG 8000 flakes, melting point 60-63 degrees. Thanks in advance, Erin Wrona San Francisco, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos - Showcase holiday pictures in hardcover Photo Books. You design it and we'll bind it! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> comcast.net Wed Jan 11 08:56:05 2006 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Wed Jan 11 08:56:15 2006 Subject: [Histonet] carbowax Message-ID: <011120061456.16144.43C51C850004AD7200003F102200750438CECE030E9D0C9A03@comcast.net> Hi Erin, There are some higher melt point paraffins available from several companies and you may wish to explore that first. Contact various manufacturers like Surgipath, Polysciences, Thermo Shandon and Richard Allan as these companies all have a variety of paraffin formulas that are designed for Histology. Most will give you some samples to try and see if it will overcome your issue. It depends on why you want the harder paraffin as to which will perform best for your laboratory. As several people have said Carbowax is a pain to use and may void warranties on some of your equipment if it is not an approved type of embedding media. This is always an area that is overlooked and is important if you are paying for repairs due to clogs or other issues rather than having it covered by a service contract or warranty. Be careful about those little details that your supervisor will not be happy with when the bill comes in even if they requested the change. Hope this helps, Pam Marcum -------------- Original message ---------------------- From: "Rittman, Barry R" > Erin > I agree totally with Rene. > Carbowax is a real pain to work with. > To harden the wax you have you may want to try adding Ceresin, a higher MP wax. > If you need a much harden wax than this then the ester waxes are more suitable > although they also have their problems. > A pertinent question here is why specifically does your lab director want or > need a higher MP wax, what problems are you having and what tissues are you > processing? > Barry > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa > Sent: Wednesday, January 11, 2006 7:54 AM > To: Erin Wrona; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] carbowax > > Hi Erin: > Many years ago Carbowax was considered "a blessing" because you could > infiltrate tissue (after fixation) with it avoiding dehydration and clearing. > This sounds great but it was a mess to store and cut because of this same > inherent quality of being water soluble (or miscible,as you wish). > If you need harder paraffin (to cut hard tissues) you just simply should try > to buy high melting point paraffin. > When I worked with plant stems I used to infiltrate them with a heigh melting > point paraffin (63-65ºC) manufactured on those days by Merck (Darmstadt). I am > sure that high melting point paraffins can be found in the market. > I thing you should avoid carbowax. > Hope this will help. > René J. > > Erin Wrona wrote: > Hi all, > > Our lab director wants us to try using carbowax polyethylene glycol as an > embedding medium because it has a slightly higher melting point than our > paraffin. I have never used it other than in formulations for frozen sections, > in which case it is water soluble. One of the other techs here thinks that it is > always water soluble. Does anyone know for sure? > > The specifics are: ems product #19770, PEG 8000 flakes, melting point 60-63 > degrees. > > Thanks in advance, > Erin Wrona > San Francisco, CA > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > --------------------------------- > Yahoo! Photos - Showcase holiday pictures in hardcover > Photo Books. You design it and we'll bind it! > _______________________________________________ > Histonet mailing list > Histonet@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 Jan 11 09:07:41 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Jan 11 09:07:53 2006 Subject: [Histonet] improper fixation In-Reply-To: Message-ID: <20060111150741.32460.qmail@web61219.mail.yahoo.com> Kristopher: After some"painful" stumbles I learned many years ago that "there is no such a thing as reprocessing a tissue". Once an improperly fixed tissue is processed, that's it! You will have to deal with the consequences. If you are talking about small biopsies the problem is even "bigger" because you risk to waste precious tissue in the intent. The best thing to do is to work with what you have and make sure it does not happen again. Ren? J. Kristopher Kalleberg wrote: All, Last week I was unable to collect 18 biopsies 2mm biopsies for a clinical that is being run and had a colleague collect them for me. After running routine H&E and Fontana Mason stains I found that two biopsies were improperly fixed. Since I have never had this problem before, I was wondering if it is possible to run the sample backwards through paraffin, xylenes, and alcohols and then fix the samples again in 10% NBF and reprocess to paraffin. If this is possible doe sanyone have a protocol or know what the effects will be by doing this? Thank you in advance. Kris Kalleberg _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. From gcallis <@t> montana.edu Wed Jan 11 09:42:11 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Jan 11 09:42:26 2006 Subject: [Histonet] Methanol and CD markers In-Reply-To: <43C43D09.1010605@u.washington.edu> References: <43C43D09.1010605@u.washington.edu> Message-ID: <6.0.0.22.1.20060111083916.01b50598@gemini.msu.montana.edu> Just a note here. Methanol is known to create problems with CD marker immunohistochemistry and that is why many use hydrogen peroxide in buffer instead of this solvent. This point was brought up in Jules Elias's book on immunohistochemistry and also on Histonet in the past. (see archives). At 04:02 PM 1/10/2006, you wrote: >We use both CD4 and CD8 on FFPE sections. We had problems with the CD4 >until we changed our quenching step to 0.5% H2O2 in MEOH for 10 min. The >pretreatment for both is 15 min microwave in pH 8 EDTA. > >Thomas Pier wrote: > >>Hello, >>I am looking for Reinhard von Wasielewski. I recently saw a post >>stating that he had working protocols for CD4 and CD8 that even worked >>in decalcified bone marrow. CD4 and CD8 in FFPE, decalcified bone >>marrow is exactly what I'm having problems with. I have been banging my >>head against the wall with this for some time now. I would really >>appreciate any help I could get on this. Thank you very much. >> >>Tom Pier >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Wed Jan 11 09:44:21 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Jan 11 09:44:34 2006 Subject: [Histonet] carbowax In-Reply-To: <20060111005222.12149.qmail@web52505.mail.yahoo.com> References: <20060111005222.12149.qmail@web52505.mail.yahoo.com> Message-ID: <6.0.0.22.1.20060111084247.01b63270@gemini.msu.montana.edu> Material safety data sheets should give this information as would a Merck index. Sometimes companies like Aldrich have this in their product description in their catalog too or EMS will tell you via tech services. At 05:52 PM 1/10/2006, you wrote: >Hi all, > > Our lab director wants us to try using carbowax polyethylene glycol as > an embedding medium because it has a slightly higher melting point than > our paraffin. I have never used it other than in formulations for frozen > sections, in which case it is water soluble. One of the other techs here > thinks that it is always water soluble. Does anyone know for sure? > > The specifics are: ems product #19770, PEG 8000 flakes, melting point > 60-63 degrees. > > T Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From sjchtascp <@t> yahoo.com Wed Jan 11 10:01:44 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Wed Jan 11 10:01:52 2006 Subject: [Histonet] TBS Polyfin Message-ID: <20060111160144.83423.qmail@web90205.mail.scd.yahoo.com> Has anyone ever used the TBS Polyfin paraffin for tissue infiltration and embedding. I'm considering a change from the paraplast I currently use. Steve --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. From glenn_krasinski <@t> yahoo.com Wed Jan 11 12:05:38 2006 From: glenn_krasinski <@t> yahoo.com (Glenn Krasinski) Date: Wed Jan 11 12:05:45 2006 Subject: [Histonet] H&E staining in solid tumors... Message-ID: <20060111180538.96519.qmail@web37109.mail.mud.yahoo.com> Can H&E staining be used to determine necrosis in solid tumors treated with agents? If yes, then would pink only staining signify areas of necrosis and purple/pink staining areas of live/intact cells? Could precent necrosis be then calculated using the areas of pink only versus purple/pink? Glenn M. Krasinski San Diego, CA --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. From sharon.osborn <@t> dnax.org Wed Jan 11 12:52:52 2006 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Wed Jan 11 12:53:28 2006 Subject: [Histonet] RE: TBS Polyfin Message-ID: <29B25753F6B1D51196110002A589D444032C496D@PALMSG30.us.schp.com> Steve, I have used TBS Polyfin for a number of years and we currently use it in this facility. I like it especially for embedding for it remain more clear when cutting. It also sets up more quickly than other paraffins when embedding so those who are more speedy in embedding are able to continue their speed with great results. It also maintains its integrity when doing block chilling and microtomy (less crazing or cracking of the paraffin than other types). It is more 'clear'or translucent than some other paraffins in allowng visualization of the specimen in the block. Cost is about the same as Paraplast (for us anyway). In some laboratories, we have used paraplast plus for the infiltration on the processors and Polyfin for the embedding without problems. I realize that some technologists feel doing this could create problems; however, we did not experience problems with the mixture in this way. Sharon Osborn DNAX, SP BioPharma Palo Alto, CA Date: Wed, 11 Jan 2006 08:01:44 -0800 (PST) From: Steven Coakley Subject: [Histonet] TBS Polyfin To: Histonet@lists.utsouthwestern.edu Message-ID: <20060111160144.83423.qmail@web90205.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Has anyone ever used the TBS Polyfin paraffin for tissue infiltration and embedding. I'm considering a change from the paraplast I currently use. Steve ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From Melissa.Gonzalez <@t> cellgenesys.com Wed Jan 11 12:53:10 2006 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Wed Jan 11 12:53:31 2006 Subject: [Histonet] GM-CSF in mouse tissue, or secreted proteins in general Message-ID: Hi all, I was wondering if anyone has ever successfully detected GM-CSF secretion in mouse tissues using immunofluorescence? (NOT on paraffin embedded sections- I have seen those papers). Do various tissues express this cytokine at different levels? Also, do secreted proteins have to be fixed and go through the whole paraffin embedding process to be properly preserved and then subsequently detected? Just curious. I always seem to find sources of staining only using chromogenic techniques on paraffin sections and the staining always seems a little suspect, kind of difficult to interpret what is background and what is real. Thanks for any input. Melissa -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, January 11, 2006 10:26 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 26, 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. Methanol and CD markers (Gayle Callis) 2. Re: carbowax (Gayle Callis) 3. TBS Polyfin (Steven Coakley) ---------------------------------------------------------------------- Message: 1 Date: Wed, 11 Jan 2006 08:42:11 -0700 From: Gayle Callis Subject: [Histonet] Methanol and CD markers To: Lori Richey , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20060111083916.01b50598@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Just a note here. Methanol is known to create problems with CD marker immunohistochemistry and that is why many use hydrogen peroxide in buffer instead of this solvent. This point was brought up in Jules Elias's book on immunohistochemistry and also on Histonet in the past. (see archives). At 04:02 PM 1/10/2006, you wrote: >We use both CD4 and CD8 on FFPE sections. We had problems with the CD4 >until we changed our quenching step to 0.5% H2O2 in MEOH for 10 min. The >pretreatment for both is 15 min microwave in pH 8 EDTA. > >Thomas Pier wrote: > >>Hello, >>I am looking for Reinhard von Wasielewski. I recently saw a post >>stating that he had working protocols for CD4 and CD8 that even worked >>in decalcified bone marrow. CD4 and CD8 in FFPE, decalcified bone >>marrow is exactly what I'm having problems with. I have been banging my >>head against the wall with this for some time now. I would really >>appreciate any help I could get on this. Thank you very much. >> >>Tom Pier >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 2 Date: Wed, 11 Jan 2006 08:44:21 -0700 From: Gayle Callis Subject: Re: [Histonet] carbowax To: Erin Wrona , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20060111084247.01b63270@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Material safety data sheets should give this information as would a Merck index. Sometimes companies like Aldrich have this in their product description in their catalog too or EMS will tell you via tech services. At 05:52 PM 1/10/2006, you wrote: >Hi all, > > Our lab director wants us to try using carbowax polyethylene glycol as > an embedding medium because it has a slightly higher melting point than > our paraffin. I have never used it other than in formulations for frozen > sections, in which case it is water soluble. One of the other techs here > thinks that it is always water soluble. Does anyone know for sure? > > The specifics are: ems product #19770, PEG 8000 flakes, melting point > 60-63 degrees. > > T Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 3 Date: Wed, 11 Jan 2006 08:01:44 -0800 (PST) From: Steven Coakley Subject: [Histonet] TBS Polyfin To: Histonet@lists.utsouthwestern.edu Message-ID: <20060111160144.83423.qmail@web90205.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Has anyone ever used the TBS Polyfin paraffin for tissue infiltration and embedding. I'm considering a change from the paraplast I currently use. Steve --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 26, Issue 11 **************************************** From c.m.vanderloos <@t> amc.uva.nl Wed Jan 11 12:54:02 2006 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Wed Jan 11 12:54:17 2006 Subject: [Histonet] RE: Histonet@lists.utsouthwestern.edu Message-ID: <456cc945c30a.45c30a456cc9@amc.uva.nl> Hi Gayle and others, Just some remarks on the methanol-issue from my perspective: Basically you are right that methanol is nasty stuff for many CD markers. Using methanol on human cryostat sections has a deleterious effect on far most antigens (yes, I am aware that the mouse-world thinks different on this). However, with FFPE sections that have been through alcohol-steps several times a methanol step doesn't seem to matter that much anymore. Once an antigen or epitope has survived embedding, methanol will not do any harm. All my FFPE sections developed for peroxidase activity are blocked with methanol + 0.3% peroxide for 20 min at RT (Streefkerk, 1972 in JHC) without any problem. Some people claim there are exceptions here, but so far I haven't seen real comparison experiments proving this. Blocking of endogenous peroxidase activity with methanol+0.3% peroxide is far more effective than PBS+03.% peroxide+0.1% sodium azide. The latter just blocks erythrocyte pseudoperoxidase but just "weaken" the endogenous peroxide activity in neutrophils. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Wed, 11 Jan 2006 08:42:11 -0700 From: Gayle Callis Subject: [Histonet] Methanol and CD markers To: Lori Richey , Histonet@lists.utsouthwestern.edu Just a note here. Methanol is known to create problems with CD marker immunohistochemistry and that is why many use hydrogen peroxide in buffer instead of this solvent. This point was brought up in Jules Elias's book on immunohistochemistry and also on Histonet in the past. (see archives). At 04:02 PM 1/10/2006, you wrote: >We use both CD4 and CD8 on FFPE sections. We had problems with the CD4 >until we changed our quenching step to 0.5% H2O2 in MEOH for 10 min. The >pretreatment for both is 15 min microwave in pH 8 EDTA. > >Thomas Pier wrote: > >>Hello, >>I am looking for Reinhard von Wasielewski. I recently saw a post >>stating tha! t he had From sharon.osborn <@t> dnax.org Wed Jan 11 13:12:07 2006 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Wed Jan 11 13:12:40 2006 Subject: [Histonet] Tissue Cassettes Message-ID: <29B25753F6B1D51196110002A589D444032C496E@PALMSG30.us.schp.com> Histoetters, We have 9 cases of Fisher Omnsette tissue cassettes with lids attached FREE to anyone willing to provide the shipping costs. Each case contains 500 cassettes with attached lids. We have not used this type for almost 2 years so rather than throw them away (non-recyclable)...perhaps someone would like them for the cost of the shipping. Please contact me to make arrangements. There are various colors in the cases. Sharon Osborn DNAX, SP BioPharma Palo Alto, CA 650-496-8834 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From Paula.F.Conlon <@t> Lahey.org Wed Jan 11 13:29:29 2006 From: Paula.F.Conlon <@t> Lahey.org (Conlon, Paula F.) Date: Wed Jan 11 13:28:40 2006 Subject: [Histonet] Xpreass processor Message-ID: Hi Histonetters, We are looking into the possibility of purchasing a microwave processor. Currently the Sakura Xpress seems to be the front runner over the Milestone. Does anyone have experience with any of them. Thanks for your input. See our web page at http://www.lahey.org for a full directory of Lahey sites, staff, services and career opportunities. THIS MESSAGE IS INTENDED FOR THE USE OF THE PERSON TO WHOM IT IS ADDRESSED. IT MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If you are not the intended recipient, your use of this message for any purpose is strictly prohibited. If you have received this communication in error, please delete the message and notify the sender so that we may correct our records. From PBugelsk <@t> CNTUS.JNJ.COM Wed Jan 11 14:26:19 2006 From: PBugelsk <@t> CNTUS.JNJ.COM (Bugelski, Peter [CNTUS]) Date: Wed Jan 11 14:26:40 2006 Subject: [Histonet] H&E staining in solid tumors... Message-ID: <7BF70FA941B9AE4783EAAF733762F1B50B3A495B@CNTUSMAEXS3.na.jnj.com> Pathologists use H&E to tell live from necrotic every day. For analysis, I'd use a mouse to draw around the necrotic areas. I would not try to get a computer to do this. You could also use the "point counting" method of stereology. You overlay an array of points on you image and counts the points on live and the number on necrotic. According to the priciples of stereology, the ratio of necrotic points:live points is proportional to % necrotic. Either way, make sure you have an adequate and random sampling. See any text on stereology for giudance. Publishing the data might be problematic. Although I consider the approach valid, since it is not "molecular" peer review could be an issue. Good luck Peter Bugelski, PhD, FRCPath -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Glenn Krasinski Sent: Wednesday, January 11, 2006 1:06 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] H&E staining in solid tumors... Can H&E staining be used to determine necrosis in solid tumors treated with agents? If yes, then would pink only staining signify areas of necrosis and purple/pink staining areas of live/intact cells? Could precent necrosis be then calculated using the areas of pink only versus purple/pink? Glenn M. Krasinski San Diego, CA --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DOOLEEO <@t> shands.ufl.edu Wed Jan 11 15:07:08 2006 From: DOOLEEO <@t> shands.ufl.edu (Elaine Dooley) Date: Wed Jan 11 15:07:19 2006 Subject: [Histonet] EGFr vIII antibody search Message-ID: Dear Histonetters, Does anyone out there know of a vendor or source for the antibody for EGFr vIII ) Epidermal Growth Factor variant three? I talked to Zymed and they discontinued it. Thanks in advance Elaine Dooley HTL Shands Hospital Gainesville FL 352-265-0111 ext 7-2114 From gcallis <@t> montana.edu Wed Jan 11 15:51:10 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Jan 11 15:51:26 2006 Subject: [Histonet] Tools aka a new website to help find antibodies and other things Message-ID: <6.0.0.22.1.20060111144420.01b52e30@gemini.msu.montana.edu> Dear all, To help find an antibody, this website may help you and was cited in the latest issue of Science. www.exactantigen.com You can search by gene, organism, and disease - and more. This may a good tool for finding some obscure or discontinued product from another source. It took some getting used to, but it worked once I played with a bit. From histosci <@t> shentel.net Wed Jan 11 16:57:34 2006 From: histosci <@t> shentel.net (HSRL) Date: Wed Jan 11 16:57:35 2006 Subject: [Histonet] GMS on frozen sections? Message-ID: <000001c61702$694179f0$0200a8c0@HSRLMAIN> Dear Netters, Does anyone have a protocol for GMS staining of OCT embedded tissues? Any help would be appreciated. Sincerely, Beth Poole HSRL, Inc.- A GLP Compliant Contract Laboratory 5930 Main Street Mount Jackson, Virginia 22842 540.477.4440 Fax: 540.477.4448 beth@hsrl.org www.hsrl.org From vazquezr <@t> ohsu.edu Wed Jan 11 17:08:39 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Wed Jan 11 17:09:14 2006 Subject: [Histonet] Tools aka a new website to help find antibodies and other things Message-ID: Thank you so much, I will check this out.... Robyn OHSU From Linresearch <@t> aol.com Wed Jan 11 17:40:13 2006 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Wed Jan 11 17:40:27 2006 Subject: [Histonet] Abs for Guinea Pig Tissues Message-ID: <29d.38c3f95.30f6f15d@aol.com> Hello, I am trying to stain Insulin, glucagon, somatostatin & cck in FFPE guinea pig. I am not having muck success with these markers which are primarily made in rabbit and using an XRb secondary. Any advice? Lin From anh2006 <@t> med.cornell.edu Wed Jan 11 21:31:00 2006 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Wed Jan 11 21:31:25 2006 Subject: [Histonet] H&E staining in solid tumors... In-Reply-To: <20060111180538.96519.qmail@web37109.mail.mud.yahoo.com> References: <20060111180538.96519.qmail@web37109.mail.mud.yahoo.com> Message-ID: This is potentially true of course but pink could also be stromal areas (sometimes nuclei so small they are hard to detect) or more relevantly areas of intense collagen/matrix deposition. Something to consider when setting something like this up. >Can H&E staining be used to determine necrosis in solid tumors >treated with agents? If yes, then would pink only staining signify >areas of necrosis and purple/pink staining areas of live/intact >cells? Could precent necrosis be then calculated using the areas of >pink only versus purple/pink? > > Glenn M. Krasinski > San Diego, CA > > >--------------------------------- >Yahoo! Photos > Got holiday prints? See all the ways to get quality prints in your >hands ASAP. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- From apagan <@t> lakewoodpath.com Thu Jan 12 08:54:43 2006 From: apagan <@t> lakewoodpath.com (apagan) Date: Thu Jan 12 08:58:07 2006 Subject: [Histonet] Excel Fixative Message-ID: <001301c61788$206b1e80$3701a8c0@lakewoodlab.com> Does anyone have information about a fixative called Excel? I do not know the vendor so that information would be helpful also. Thanks, Audrey Audrey Pagan Lakewood Pathology Associates 1200 River Ave. Suite 10E Lakewood, NJ 08701 phone: 732-901-7575 ext. 102 fax: 732-901-9482 From sluhisto <@t> yahoo.com Thu Jan 12 09:07:37 2006 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Thu Jan 12 09:07:48 2006 Subject: [Histonet] Automated Special Stains Message-ID: <20060112150737.73873.qmail@web51004.mail.yahoo.com> Hello All: Can you share with me, those of you who have automated your special stains, what instrument you have chosen, what ones you looked at, and why you chose the one you did? We are heading that direction and I would like very much to tap your expertise. Thanks, in advance, and have a super weekend. Susan Histopathology Supervisor St. Louis University Medical School --------------------------------- Yahoo! Photos ? Showcase holiday pictures in hardcover Photo Books. You design it and we?ll bind it! From ROrr <@t> enh.org Thu Jan 12 09:53:30 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Thu Jan 12 09:53:39 2006 Subject: [Histonet] Herceptest Message-ID: Hello everyone, Could you please let me know if you are using the Herceptest kit or the Herceptest antibody and seeing any staining in the negative epithelials? I would be interested in any response by anyone using a polyclonal antibody for Her2neu. I understand this could be appropriate staining and should be subtracted off the evaluation of intensity. If you do see this, would you call it ?+ or even 1+ in your lab? Thank-you for your reply! Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 From pathrm35 <@t> adelphia.net Thu Jan 12 10:16:22 2006 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Thu Jan 12 10:16:30 2006 Subject: [Histonet] Automated Special Stains Message-ID: <21322996.1137082582854.JavaMail.root@web16> Susan, We use the Biogenex IHC stainer for specials and IHC, which can be run together. We use mostly PASF and Alcian Blue/PAS stains as we only perform dermpath procedures but Biogenex has many other stains. For the cost of the stainer, the good results and good service I would recommend Biogenex. Ron Martin ---- Histology SLU wrote: > Hello All: > > Can you share with me, those of you who have automated your special stains, what instrument you have chosen, what ones you looked at, and why you chose the one you did? We are heading that direction and I would like very much to tap your expertise. Thanks, in advance, and have a super weekend. > > Susan > Histopathology Supervisor > St. Louis University Medical School > > > > --------------------------------- > Yahoo! Photos ? Showcase holiday pictures in hardcover > Photo Books. You design it and we?ll bind it! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Jan 12 12:33:44 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Jan 12 12:33:53 2006 Subject: [Histonet] odefreitasIHCRG Membership Application Form Message-ID: <200601121833.k0CIXgwZ024821@chip.viawest.net> verify NSh membership please Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _____ From: webmaster@neo.agsci.colostate.edu [mailto:webmaster@neo.agsci.colostate.edu] Sent: Thursday, January 12, 2006 10:54 AM To: pruegg@ihctech.net Subject: IHCRG Membership Application Form _____ Last_Name: DeFreitas First_Name: Otis NSH_Member: Yes Employer: our lady of lourdes & Virtua Address: 1600 Haddon avenue City: Camden State: NJ Zip: 08103 Province: Country: Phone: 856-757-3584 Ext: Fax: Email: redemptionsharps@yahoo.com Surgical_Pathology: Yes Hematopathology: Orthopedic: Veterinary: Immunology: Plastics: Research: Paraffin: Yes Frozen: Yes Cytospins: Yes Smears_Touch_Preps: Plastics_sample: Sample_Other: PAP: APAAP: ABC_HRP: Yes ABC_AP: Yes SA_HRP: Yes SA_AP: IF: IHC_Other: ISH: Yes Gels: PCR: Mol_Other: Automated_IHC: Yes Company: Dako IHC_Qualification: No Date_Taken_Exam: Like_To_Take_Exam: Yes Expected_Date: 7/2006 Need_Tissues: No Tissue_Needed: Need_Reagents: No Reagents_Needed: Can_Provide_Tissues: No Tissues_Provided: Can_Provide_Reagents: No Reagents_Provided: Form: Submit Remote Name: 198.245.165.200 HTTP User Agent: Mozilla/4.0 (compatible; MSIE 6.0; Windows NT 4.0) Other_Tech From pruegg <@t> ihctech.net Thu Jan 12 13:00:24 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Jan 12 13:00:35 2006 Subject: [Histonet] disregard Message-ID: <200601121900.k0CJ0NwZ002454@chip.viawest.net> Sorry folks, I hit the wrong button and sent a message to histonet that was meant for NSH, please disregard "verify membership" message. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From hymclab <@t> hyhc.com Thu Jan 12 13:20:42 2006 From: hymclab <@t> hyhc.com (hymclab) Date: Thu Jan 12 13:19:06 2006 Subject: [Histonet] Microwave slide drying vs. conventional slide dryi ng Message-ID: I have been drying slides in the microwave since I came here in 1992. All slides for all applications (H&E, specials, immunos, etc...) are dried in a regular kitchen microwave for 6 minutes. The only draw back I have found is that we have to use plastic racks to dry them in and then transfer to our metal staining rack for our autostainer (our autostainer does not have a drying station built in). What we do to make sure there is consistent drying is to have a urine cup full of water sitting next to the drying rack with the slides. We always get conisistent drying and have no artifact. We have sent some our slides out for consultation and have gotten back comments that our slides are excellent quality. I have a new Histo grade microwave sitting here, but have not calculated the time needed to dry our slides yet, but will shortly. I have been flamed before for stating that I use the microwave for slide drying, so please save your flaming!!!!! Some of us just do what works!!!!! Dawn D. Schneider, HT(ASCP) Lead Histology Tech LAB/LIS Coordinator Howard Young Medical Center 240 Maple Ave. Woodruff, WI 54568 ?(715) 356-8174 -----Original Message----- From: lharris@samhealth.org [mailto:lharris@samhealth.org] Sent: Friday, January 06, 2006 10:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave slide drying vs. conventional slide drying We are having a discussion about slide drying at out facility. How many of you out there use a microwave to dry your H&E slides before staining versus using a conventional 70 degree Celsius hot air slide drying oven for 20 minutes? I personally against microwave slide drying but the doctors like it because it takes less time (3 min. 30 sec. on medium setting). Thanks for your response. Lori A. Harris, HT (ASCP) Histology Section Leader GSRMC - Pathology 3600 NW Samaritan Drive Corvallis, OR 97330 1-541-768-6078 lharris@samhealth.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 From vazquezr <@t> ohsu.edu Thu Jan 12 13:39:30 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Jan 12 13:40:01 2006 Subject: [Histonet] odefreitasIHCRG Membership Application Form Message-ID: What is this? Robyn OHSU From m.doube <@t> qmul.ac.uk Thu Jan 12 14:34:50 2006 From: m.doube <@t> qmul.ac.uk (Michael Doube) Date: Thu Jan 12 14:35:01 2006 Subject: [Histonet] paraformaldehyde in the microwave Message-ID: <43C6BD6A.1050103@qmul.ac.uk> Hi there It's been a while since I last made 4% paraformaldehyde in phosphate-buffered saline - it was a few years back as an undergrad. This time I'm in a new lab, and found that there is no hotplate! So, after trying to keep the suspension warm with hot water baths (tedious) , I resorted to the microwave, on low power. It seems to work much faster than what I remember with the conventional technique. Has anyone had any experience with this? Mike -- Michael Doube BPhil BVSc MRCVS MPhil / PhD Student Dental Institute Barts and The London School of Medicine and Dentistry Queen Mary, University of London New Rd London E1 1BB United Kingdom Phone +44 (0)20 7377 7000 ext 2681 From pruegg <@t> ihctech.net Thu Jan 12 15:01:10 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Jan 12 15:01:31 2006 Subject: [Histonet] odefreitasIHCRG Membership Application Form In-Reply-To: Message-ID: <200601122101.k0CL1AAq008309@chip.viawest.net> Sorry I sent this to the wrong place. Please disregard. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] Sent: Thursday, January 12, 2006 12:40 PM To: pruegg@ihctech.net; histonet@pathology.swmed.edu Subject: Re: [Histonet] odefreitasIHCRG Membership Application Form What is this? Robyn OHSU From KMB1904 <@t> aol.com Thu Jan 12 15:54:37 2006 From: KMB1904 <@t> aol.com (KMB1904@aol.com) Date: Thu Jan 12 15:54:55 2006 Subject: [Histonet] mucicarmine in the microwave?? Message-ID: <258.50c8ab7.30f82a1d@aol.com> I have a tech that is telling me mucicarmine can be done in the microwave...Is this true? I didnt think it was a good idea to microwave mucicarmine. Any procedures..or thoughts would be great! From rjbuesa <@t> yahoo.com Thu Jan 12 16:52:58 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Jan 12 16:53:09 2006 Subject: [Histonet] mucicarmine in the microwave?? In-Reply-To: <258.50c8ab7.30f82a1d@aol.com> Message-ID: <20060112225258.28499.qmail@web61221.mail.yahoo.com> You can greatly reduce the time if after blueing the hematoxylin you heat the slides in mucicarmin in a MW to reach 60?C and keep the slides at that temp. x 10 min. The MW should have a temperature probe or you have to keep adding energy bursts to keep the temp. a 60?C At the end of the 10 min wash quickly with dist. water → 95% ethanol → light green for 1 min. → dehydrate → clear → mount. If your MW oven does not have a temp. probe, you can heat the slides in 50 mL of the mucicarmin sol. for 35 sec.s, take the slides to a water bath at 60?C and repeat the heating step every 5 mins. during a total of 20 min. (or more, depending on the "age" of the mucicarmin sol. Ren? J. KMB1904@aol.com wrote: I have a tech that is telling me mucicarmine can be done in the microwave...Is this true? I didnt think it was a good idea to microwave mucicarmine. Any procedures..or thoughts would be great! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos ? Showcase holiday pictures in hardcover Photo Books. You design it and we?ll bind it! From AFoshey <@t> chw.org Thu Jan 12 17:38:01 2006 From: AFoshey <@t> chw.org (Foshey, Annette) Date: Thu Jan 12 17:38:14 2006 Subject: [Histonet] Job opening!!! Message-ID: <9E6D52F532809247BDA1783680E92C56586882@CHWEXC.chwi.chswi.org> Children's Hospital and Health System is looking for a full-time Histology Technician. Responsibilities include, processing, embedding, cutting and staining. Knowledge of operation of Artisan stainer and Impac AP system a plus. Hours include Saturday rotation. New Grads welcome! If interested please apply on line at: www.chw.org Annette Foshey, HT Team Leader in Histology Children's Hospital of Wisconsin ************************** Children's Hospital and Health System recognizes that unencrypted e-mail is insecure and does not guarantee confidentiality. The confidentiality of replies to this message cannot be guaranteed unless the replies are encrypted. From wilson_jm <@t> cimar.org Thu Jan 12 19:05:25 2006 From: wilson_jm <@t> cimar.org (Jonathan Wilson) Date: Thu Jan 12 19:05:48 2006 Subject: [Histonet] non-specific cross reactivity with mucus References: <19df38c19ddd3a.19ddd3a19df38c@amc.uva.nl> Message-ID: <004501c617dd$72eec620$c05ca93e@COBITIS> Hello, I work with fish in a resarch lab and with some of the antibodies I use I get what I would consider non-specific crossreactivity with mucus either covering the body surface or in mucocyte granules. More often this problem would arise with rabbit polyclonal anti-peptide antibodies using neat serum. I usually get around this by affinity purifying using the peptide but in some cases this is not possible. Someone once told me that mucoproteins have a large number of antigenic determinants increasing the likelyhood of crossreactivity but I could never find a reference. I've searched through the archives but couldn't find an explanation except for a suggestion that it might be charge related. Could someone please shed some light on this situation for me. It is something that has bugged me for the longest time. Thank you for any insight. Sincerely, Jon From conniegrubaugh <@t> hotmail.com Thu Jan 12 19:39:37 2006 From: conniegrubaugh <@t> hotmail.com (connie grubaugh) Date: Thu Jan 12 19:39:47 2006 Subject: [Histonet] Disciplinary plan Message-ID: We need any and all suggestions to implement a disciplinary plan for slide labeling errors. 99% of the time the errors are caught by the person that is labeling the slides and checking them with the blocks and reports before they leave the lab. But we have a on going problem with one tech that mislabeled slides or does not label them at all. We process around 400 blocks a day and on a regular day there can be any where from 2-8 cases of slides that are mislabeled. Since we are a reference lab and have drivers waiting to take the slides to the doctors at hospitals around the valley this has slowed us up. Also for improper embedding of specimens. Obvious specimens like vas deferens and cervical cones. Since we have a assistant doing the labeling and putting the cases together for send out she is not the familiar with what all tissues are supposed to look like. Thanks. Connie Grubaugh Las Vegas Nv. From cbass <@t> bidmc.harvard.edu Thu Jan 12 20:16:25 2006 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Thu Jan 12 20:16:44 2006 Subject: [Histonet] antibodies to distinguish neurons and glia Message-ID: Hi Guys, I have a protein being expressed in the brain and I would like to see if it is in neurons or glial cells. Could someone recommend some good antibodies for this? I think GFAP and NeuN are considered the standard. I just need a reliable source for these. If you have any suggestions as to dilutions and incubations times that would be great. This is paraformaldehyde fixed mouse brain, 30 micron floating sections. I am planning to do double fluorescent immunostaining. Thanks, Caroline Bass From jkiernan <@t> uwo.ca Fri Jan 13 00:54:02 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Jan 13 00:54:13 2006 Subject: [Histonet] paraformaldehyde in the microwave References: <43C6BD6A.1050103@qmul.ac.uk> Message-ID: <43C74E8A.7D4DCC5@uwo.ca> The source of heat does not matter. Paraformaldehyde depolymerizes at 60C in the presence of a base such as disodium hydrogen phosphate. It won't depolymerize ("dissolve") when heated in pure water. This information is present in every histotechnical textbook published since about 1960. The chemistry is very simple, and abundantly available on the internet. Paraformaldehyde and formalin are sources of hydrated formaldehyde, which fixes tissues well if used at approximately neutral pH for 12-24 hours. John Kiernan London, Canada. ----------------------------------- Michael Doube wrote: > > Hi there > > It's been a while since I last made 4% paraformaldehyde in > phosphate-buffered saline - it was a few years back as an undergrad. > This time I'm in a new lab, and found that there is no hotplate! So, > after trying to keep the suspension warm with hot water baths (tedious) > , I resorted to the microwave, on low power. It seems to work much > faster than what I remember with the conventional technique. Has anyone > had any experience with this? > > Mike > > -- > Michael Doube BPhil BVSc MRCVS > MPhil / PhD Student > Dental Institute > Barts and The London School of Medicine and Dentistry > Queen Mary, University of London > New Rd > London E1 1BB > United Kingdom > > Phone +44 (0)20 7377 7000 ext 2681 From louise.renton <@t> gmail.com Fri Jan 13 01:04:02 2006 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Jan 13 01:04:10 2006 Subject: [Histonet] Disciplinary plan In-Reply-To: References: Message-ID: Connie, This is my take on the problem: Whereas I agree that discipline might be one avenue to correct your problem, it might be important to also bear in mind that many so called "errors" are based on poor understanding of the processes and importance of the task. Instead of "cracking the whip", education as to the dire effects of mislabelling and involvement of the problem individual in solving the problem itself can be of assistance. Assuming that all employees have the same priorities as management is dangerous as is the assumption that all histotechs are created equal and aresufficiently experienced to be readily able to distinguish a vas deferns from a non-descript piece of tissue at a glance. It sounds like your lab is a fairly high pressured set-up, keep in mind that not all individuals are geared to work at the same pace with the same degee of accuracy. They might well work better at a slightly less pressured pace. If you want a maximal degree of consistency hire a robot! (On the other hand, perhaps your tech is bored and needs something more challenging). So my suggestion are: Try to: 1. Determine why there is a problem with this tech - and whether they are long term (attention deficit, poor numeracy) or not (sometimes poor work performance is related to personal issues) 2. Educate and explain HOW things are done in your lab, WHY they are done just so and WHAT the consequences are of failing to do thenm the right way. Re train if necessary 3. Offer alternative duties - less cutting & labelling, more embedding, or alternatively more challenge. These are my thoughts and are not in any way intended to impugn your management style Best regards On 1/13/06, connie grubaugh wrote: > We need any and all suggestions to implement a disciplinary plan for slide > labeling errors. 99% of the time the errors are caught by the person that > is labeling the slides and checking them with the blocks and reports before > they leave the lab. But we have a on going problem with one tech that > mislabeled slides or does not label them at all. We process around 400 > blocks a day and on a regular day there can be any where from 2-8 cases of > slides that are mislabeled. Since we are a reference lab and have drivers > waiting to take the slides to the doctors at hospitals around the valley > this has slowed us up. Also for improper embedding of specimens. Obvious > specimens like vas deferens and cervical cones. > Since we have a assistant doing the labeling and putting the cases together > for send out she is not the familiar with what all tissues are supposed to > look like. > > > Thanks. > Connie Grubaugh > Las Vegas Nv. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....He was a very conceited cowboy, he only rode on pompous grass. From Lobke.DeBels <@t> UGent.be Fri Jan 13 04:59:39 2006 From: Lobke.DeBels <@t> UGent.be (Lobke De Bels) Date: Fri Jan 13 04:59:53 2006 Subject: [Histonet] fixation and embedding eyeball Message-ID: <1137149979.43c7881b11a02@mail.ugent.be> Dear all, we have some problems with the fixation and/or embedding of an eyeball (dog), the different structures don't stay together. Now we wanna try celloidin embedding but we don't have any experience with that technique. Could you give us some information to start the celloidin embedding technique? who has experience with fixation and embedding of whole eyeballs? Many thanks in advance for any help Lobke De Bels Department of Morphology Faculty of Veterinary Medicine Ghent University Salisburylaan 133 9820 Merelbeke Belgium lobke.debels@ugent.be -- From m.doube <@t> qmul.ac.uk Fri Jan 13 05:27:05 2006 From: m.doube <@t> qmul.ac.uk (Michael Doube) Date: Fri Jan 13 05:27:15 2006 Subject: [Histonet] paraformaldehyde in the microwave In-Reply-To: <43C74E8A.7D4DCC5@uwo.ca> References: <43C6BD6A.1050103@qmul.ac.uk> <43C74E8A.7D4DCC5@uwo.ca> Message-ID: <43C78E89.9000500@qmul.ac.uk> Thanks for your reply John. I am aware that certain tissue prep protocols are substantitally accelerated by microwave radiation independent of a direct thermal effect, so my question was more to discover whether a similar acceleration effect might operate duing simple chemical preparation. This is based on the chance observation that a few minutes on low, with the PFA suspension in a flask in a large water bath was sufficient to make my PFA depolymerise. Thanks to all those concerned about fume safety, yes we are fume hooded and well ventilated (to a fault, actually) and I seal the top of my flask. Michael John Kiernan wrote: >The source of heat does not matter. >Paraformaldehyde depolymerizes at 60C in the >presence of a base such as disodium hydrogen >phosphate. It won't depolymerize ("dissolve") when >heated in pure water. > >This information is present in every >histotechnical textbook published since about >1960. The chemistry is very simple, and abundantly >available on the internet. Paraformaldehyde and >formalin are sources of hydrated formaldehyde, >which fixes tissues well if used at approximately >neutral pH for 12-24 hours. > >John Kiernan >London, Canada. > > >----------------------------------- >Michael Doube wrote: > > >>Hi there >> >>It's been a while since I last made 4% paraformaldehyde in >>phosphate-buffered saline - it was a few years back as an undergrad. >>This time I'm in a new lab, and found that there is no hotplate! So, >>after trying to keep the suspension warm with hot water baths (tedious) >>, I resorted to the microwave, on low power. It seems to work much >>faster than what I remember with the conventional technique. Has anyone >>had any experience with this? >> >>Mike >> >>-- >>Michael Doube BPhil BVSc MRCVS >>MPhil / PhD Student >>Dental Institute >>Barts and The London School of Medicine and Dentistry >>Queen Mary, University of London >>New Rd >>London E1 1BB >>United Kingdom >> >>Phone +44 (0)20 7377 7000 ext 2681 >> >> -- Michael Doube BPhil BVSc MRCVS MPhil / PhD Student Dental Institute Barts and The London School of Medicine and Dentistry Queen Mary, University of London New Rd London E1 1BB United Kingdom Phone +44 (0)20 7377 7000 ext 2681 From BMolinari <@t> heart.thi.tmc.edu Fri Jan 13 05:38:19 2006 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Jan 13 05:43:44 2006 Subject: [Histonet] Disciplinary plan Message-ID: One more suggestion, we had this problem and the tech had her eyes checked and found she needed glasses. The result in the lab was immediate. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise renton Sent: Friday, January 13, 2006 1:04 AM To: connie grubaugh; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Disciplinary plan Connie, This is my take on the problem: Whereas I agree that discipline might be one avenue to correct your problem, it might be important to also bear in mind that many so called "errors" are based on poor understanding of the processes and importance of the task. Instead of "cracking the whip", education as to the dire effects of mislabelling and involvement of the problem individual in solving the problem itself can be of assistance. Assuming that all employees have the same priorities as management is dangerous as is the assumption that all histotechs are created equal and aresufficiently experienced to be readily able to distinguish a vas deferns from a non-descript piece of tissue at a glance. It sounds like your lab is a fairly high pressured set-up, keep in mind that not all individuals are geared to work at the same pace with the same degee of accuracy. They might well work better at a slightly less pressured pace. If you want a maximal degree of consistency hire a robot! (On the other hand, perhaps your tech is bored and needs something more challenging). So my suggestion are: Try to: 1. Determine why there is a problem with this tech - and whether they are long term (attention deficit, poor numeracy) or not (sometimes poor work performance is related to personal issues) 2. Educate and explain HOW things are done in your lab, WHY they are done just so and WHAT the consequences are of failing to do thenm the right way. Re train if necessary 3. Offer alternative duties - less cutting & labelling, more embedding, or alternatively more challenge. These are my thoughts and are not in any way intended to impugn your management style Best regards On 1/13/06, connie grubaugh wrote: > We need any and all suggestions to implement a disciplinary plan for slide > labeling errors. 99% of the time the errors are caught by the person that > is labeling the slides and checking them with the blocks and reports before > they leave the lab. But we have a on going problem with one tech that > mislabeled slides or does not label them at all. We process around 400 > blocks a day and on a regular day there can be any where from 2-8 cases of > slides that are mislabeled. Since we are a reference lab and have drivers > waiting to take the slides to the doctors at hospitals around the valley > this has slowed us up. Also for improper embedding of specimens. Obvious > specimens like vas deferens and cervical cones. > Since we have a assistant doing the labeling and putting the cases together > for send out she is not the familiar with what all tissues are supposed to > look like. > > > Thanks. > Connie Grubaugh > Las Vegas Nv. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....He was a very conceited cowboy, he only rode on pompous grass. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lenaspencer <@t> insightbb.com Fri Jan 13 07:56:36 2006 From: lenaspencer <@t> insightbb.com (lena spencer) Date: Fri Jan 13 07:51:00 2006 Subject: [Histonet] Job Opportunity Message-ID: <001f01c61849$2bcd6c80$3379df0c@org.insightbb.com> Norton Healthcare, Louisville, KY Histologist Position Open Full Time/Day Shift Occasional week-ends Duties: embedding, sectioning, performing special stains, frozen sections and assisting at surgical bench. Contact: Mary Beth Knight - 502-629-7884 or send resume' to marybeth.knight@nortonhealthcare.org ----- Original Message ----- From: To: Sent: Thursday, January 12, 2006 1:10 PM Subject: Histonet Digest, Vol 26, Issue 12 > This message has been processed by Symantec's AntiVirus Technology. > > Unknown00000000.data was not scanned for viruses because too many nested levels of files were found. > > > For more information on antivirus tips and technology, visit > http://ses.symantec.com/ From DeBrosse_Beatrice <@t> Allergan.com Fri Jan 13 09:24:01 2006 From: DeBrosse_Beatrice <@t> Allergan.com (DeBrosse_Beatrice) Date: Fri Jan 13 09:24:40 2006 Subject: [Histonet] fixation and embedding eyeball Message-ID: What kind of fixative are you using? We use Davidson's fixative for our eyes (including dogs), embed them in paraffin, cut them at 4 microns and the structure of the eye stays together beautifully. Good luck! Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lobke De Bels Sent: Friday, January 13, 2006 3:00 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fixation and embedding eyeball Dear all, we have some problems with the fixation and/or embedding of an eyeball (dog), the different structures don't stay together. Now we wanna try celloidin embedding but we don't have any experience with that technique. Could you give us some information to start the celloidin embedding technique? who has experience with fixation and embedding of whole eyeballs? Many thanks in advance for any help Lobke De Bels Department of Morphology Faculty of Veterinary Medicine Ghent University Salisburylaan 133 9820 Merelbeke Belgium lobke.debels@ugent.be -- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Jan 13 09:51:53 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Jan 13 09:52:08 2006 Subject: [Histonet] Disciplinary plan In-Reply-To: References: Message-ID: <6.0.0.22.1.20060113084257.01b4b7a0@gemini.msu.montana.edu> Betsy, Excellent suggestion. Safety glasses with bifocal magnification on the safety lenses that fit OVER prescription glasses are available from Fisher, Marketlab, Newcomer Supply. We use these and the techs have their own for their personal use. It is a cheap way to help out at embedding center or other hard to see tasks, even microtomy. Or make sure there is good lighting and a decent magnifier on embedding center. We prefer the bifocals instead - they don't get in the way. For labeling problems in a high through-put reference lab, consider buying a slide labeler. Writing on slides is often messy and hard to read, the same goes for cassettes. These instruments also eliminate repetitive motion problems on labeling slanted surfaces and tons of slides. At 04:38 AM 1/13/2006, you wrote: >One more suggestion, we had this problem and the tech had her eyes >checked and found she needed glasses. The result in the lab was >immediate. > >Betsy Molinari HT (ASCP) Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Fri Jan 13 10:02:17 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Jan 13 10:02:32 2006 Subject: [Histonet] RE: fixation and embedding eyeball In-Reply-To: References: Message-ID: <6.0.0.22.1.20060113085230.01b7a5e0@gemini.msu.montana.edu> Beatrice, What paraffin are you using, water bath temperature? Eyes sometimes explode on a waterbath, or do not flatten correctly. Any hints on those issues? Also, Mary Georger is an expert in eye histotechniques as she taught ab excellent hands on workshop some years ago at NSH convention. If she is not on Histonet, maybe she would elaborate privately and I could pass this on. Davidson's is a popular fixative for eyes, although she used NBF, Bouins has be suggested . With the latter, some do not like the picric acid additive. The choice of paraffin often helps - Peel a Way is one. Years ago, AFIP used Peelaway for eyes, and the person doing this explained why this paraffin was superior for eyes. If I recall our discussion correctly, it is because the paraffin doesn't expand as much as other paraffins and holds eye structures in place better. Would anyone entertain discussion on the physical properties of Peel a Way paraffin? It used to come in range of melting points, but not sure what is supplied now. At 08:24 AM 1/13/2006, you wrote: >What kind of fixative are you using? We use Davidson's fixative for our >eyes (including dogs), embed them in paraffin, cut them at 4 microns and >the structure of the eye stays together beautifully. Good luck! > >Beatrice DeBrosse-Serra Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From rjbuesa <@t> yahoo.com Fri Jan 13 10:11:43 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Jan 13 10:11:52 2006 Subject: [Histonet] Disciplinary plan In-Reply-To: Message-ID: <20060113161143.33292.qmail@web61212.mail.yahoo.com> Dear Connie: I am concerned about your posting. On the one hand it is a grave mistake if undetected because it can affect patient care. On the other hand I think that you should try to determine the cause of the problem befora taking any action. 1- was the HT trained correctly to begin with? 2- has this type of mistake by her being a long standing issue or is it something new? 3- if is a long standing problems, why it has been allowed to continue? 4- what has been done, talking with her, to find if there is a personal underlying cause? 5- if it is something new, does she have a personal problem that makes her make more mistakes now than before? Your mission is not that of a psichyatrists or social worker, your mission is to provide a work environment conducent to a work of high quality and to provide to your staff with what they need to accomplish that goal. This means that you don't have to go around trying to get involved in their personal lives (this would be extremely counterproductive) but you have to be sure to identify problems to address them correctly. When you have found the cause of the problem, then you should try to solve it (if it is related to the job only). Once you have done that then you can begin taking disciplinary actions under the guidance of your Human Resources department that for sure has a protocol. Retraining, verbal and written counselings that could lead to temporary suspension or even termination are all steps that could be taken if deemed appropriate and admisible and only if you have very well defined competencies and standards of performance, known to all and acknowledged as received to be followed. Now a piece of advise: never make an employee to feel dreadful or to curse the moment that she or he has to go to work. This will only worsen performance and will also create a umpleasant environment. Please excuse me this very long answer but it is the case that "I have been there" and "I have done that". Hope this will help you (and your histotech as well)! Ren? J. connie grubaugh wrote: We need any and all suggestions to implement a disciplinary plan for slide labeling errors. 99% of the time the errors are caught by the person that is labeling the slides and checking them with the blocks and reports before they leave the lab. But we have a on going problem with one tech that mislabeled slides or does not label them at all. We process around 400 blocks a day and on a regular day there can be any where from 2-8 cases of slides that are mislabeled. Since we are a reference lab and have drivers waiting to take the slides to the doctors at hospitals around the valley this has slowed us up. Also for improper embedding of specimens. Obvious specimens like vas deferens and cervical cones. Since we have a assistant doing the labeling and putting the cases together for send out she is not the familiar with what all tissues are supposed to look like. Thanks. Connie Grubaugh Las Vegas Nv. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From HParker <@t> Skaggs.Net Fri Jan 13 10:42:11 2006 From: HParker <@t> Skaggs.Net (Parker, Helayne) Date: Fri Jan 13 10:42:34 2006 Subject: [Histonet] Eosin too pink Message-ID: <2FABC6145388F24EBA8CBD6EC165BCCB02AFB9E5@mail1-schc.skaggs.net> Hi List, We are currently doing our H & E stain by hand and using the Richard-Allan 7211 Hematoxylin and their Eosin Y. Our pathologist is still complaining of slides being TOO PINK. I have cut the eosin step down to just 5 dips from 15 dips, and still too pink. The Eosin is very rich. This is our protocol: Hydrate to water like normal (thru Clear Rite 3, 100% and 95% Flex) Hematoxylin- 2 mins Rinsing in water until the water runs clear Clarifier II- 30 dips running water -30 dips Richard Allan Bluing - 30 dips running water - 30 dips 95% Flex - 10 dips Eosin-Y- 5 quick dips Thru 95%, 100% etc to xylene Any suggestions on a possible method change or a more mild Eosin. Complaint seems to solely "TOO PINK". I have done some experimenting w/ cutting the eosin w/ 80% Flex- and it does calm it down a bit but, the Hemo looks more purply then blue, almost periwinkle so not sure if that will be better or not- we still need to have him review the test slides. Any suggestions ? Helayne Parker Histology Section Head Skaggs Community Health Center Branson, Missouri From Lee.Ridley <@t> nottingham.ac.uk Fri Jan 13 10:48:57 2006 From: Lee.Ridley <@t> nottingham.ac.uk (Lee Ridley) Date: Fri Jan 13 10:49:34 2006 Subject: [Histonet] Pax-3 Message-ID: Dear All, Has anyone had any experience of using a Pax-3 antibody on FFPE tissue sections. I am experiencing a problem with my Antibody from Abcam and would be really grateful to hear from anyone that has success using this antibody and what you used as a positive control tissue Hope someone can help With Kindest Regards Lee Lee Ridley Research Associate The Children's Brain Tumour Research Centre University of Nottingham The Medical School (Floor D) Nottingham NG7 2UH Email: Lee.ridley@nottingham.ac.uk Tel: 0115 8230718 This message has been checked for viruses but the contents of an attachment may still contain software viruses, which could damage your computer system: you are advised to perform your own checks. Email communications with the University of Nottingham may be monitored as permitted by UK legislation. From DeBrosse_Beatrice <@t> Allergan.com Fri Jan 13 10:58:25 2006 From: DeBrosse_Beatrice <@t> Allergan.com (DeBrosse_Beatrice) Date: Fri Jan 13 10:59:36 2006 Subject: [Histonet] RE: fixation and embedding eyeball Message-ID: Hi Gayle, We use Paraplast Plus with a melting point at 56 degrees Celsius; the water temperature is what we use for other tissues, which is around 44 and 48 degrees Celsius. Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Friday, January 13, 2006 8:02 AM To: DeBrosse_Beatrice; Histonet@lists.utsouthwestern.edu Cc: Georger, Mary Subject: RE: fixation and embedding eyeball Beatrice, What paraffin are you using, water bath temperature? Eyes sometimes explode on a waterbath, or do not flatten correctly. Any hints on those issues? Also, Mary Georger is an expert in eye histotechniques as she taught ab excellent hands on workshop some years ago at NSH convention. If she is not on Histonet, maybe she would elaborate privately and I could pass this on. Davidson's is a popular fixative for eyes, although she used NBF, Bouins has be suggested . With the latter, some do not like the picric acid additive. The choice of paraffin often helps - Peel a Way is one. Years ago, AFIP used Peelaway for eyes, and the person doing this explained why this paraffin was superior for eyes. If I recall our discussion correctly, it is because the paraffin doesn't expand as much as other paraffins and holds eye structures in place better. Would anyone entertain discussion on the physical properties of Peel a Way paraffin? It used to come in range of melting points, but not sure what is supplied now. At 08:24 AM 1/13/2006, you wrote: >What kind of fixative are you using? We use Davidson's fixative for our >eyes (including dogs), embed them in paraffin, cut them at 4 microns and >the structure of the eye stays together beautifully. Good luck! > >Beatrice DeBrosse-Serra Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From sharon.osborn <@t> dnax.org Fri Jan 13 11:21:44 2006 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Jan 13 11:22:20 2006 Subject: [Histonet] RE: Fisher Omnicassettes Message-ID: <29B25753F6B1D51196110002A589D444032C4978@PALMSG30.us.schp.com> Histonetters! What a response! Thank you! Many of you emailed or tried to telephone about wanting the cassettes or suggestions for them. I apologize for inadvertently combining my lab and home number to print an invalid one. Perhaps the incorrect phone number served a greater purpose since the Henry Cogswell Community College of Everett, WA found a way to get the correct phone number and contacted me directly. We are in the process of working out the shipping details to get the 9 cases (4500 cassettes and lids) to them. Plus, we have some slides, coverslips, etc that will be included. So, again, thanks for the response! And, do you have coverslips, slides, staining dishes and slide carriers, embedding centers, IHC machines, processors, microtomes, blades, cassette and slide labelers etc. that you no longer use and are squirreled away in a drawer, back room, storage, etc that you could resurrect and donate to the college? I have been in hstology long enough to know this is often the case--so rather than trash these items, donate! Shipping is sometimes an issue,,,see if your organization will pay and take a tax deduction... some will, some won't--tis worth a try! Then work to figure out another way to pay for shipping. I suggest you contact Larry Boyd directly to see what they need, desire, wish, could use...his contact information is: email: l.boyd@henrycogswell.edu. Larry D. Boyd Lecturer-Microbiology ad Assistat to the President for Bio-Medcal Scences Henry Cogswell College Peed Hall 3002 Colby Ave Everett, WA 98201 phone: 425.258-3351 ext 106 To your wealth and your health! Sharon Osborn DNAX, SP BioPharma Palo Alto, CA 650-496-6539 *the correct phone! ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From Terry.Marshall <@t> rothgen.nhs.uk Fri Jan 13 11:54:18 2006 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Jan 13 11:53:03 2006 Subject: [Histonet] Eosin too pink Message-ID: Wouldn't we all *love* to see just what is considered too pink:-) An old pathologist friend of mine used to put a pinch of bicarb in the eosin when the techs were not looking. He had a thing of not liking it too pink. For me, I can't have it too pink, as long as the nuclei are blue. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Parker, Helayne [mailto:HParker@Skaggs.Net] Sent: 13 January 2006 16:42 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Eosin too pink Hi List, We are currently doing our H & E stain by hand and using the Richard-Allan 7211 Hematoxylin and their Eosin Y. Our pathologist is still complaining of slides being TOO PINK. I have cut the eosin step down to just 5 dips from 15 dips, and still too pink. The Eosin is very rich. This is our protocol: Hydrate to water like normal (thru Clear Rite 3, 100% and 95% Flex) Hematoxylin- 2 mins Rinsing in water until the water runs clear Clarifier II- 30 dips running water -30 dips Richard Allan Bluing - 30 dips running water - 30 dips 95% Flex - 10 dips Eosin-Y- 5 quick dips Thru 95%, 100% etc to xylene Any suggestions on a possible method change or a more mild Eosin. Complaint seems to solely "TOO PINK". I have done some experimenting w/ cutting the eosin w/ 80% Flex- and it does calm it down a bit but, the Hemo looks more purply then blue, almost periwinkle so not sure if that will be better or not- we still need to have him review the test slides. Any suggestions ? Helayne Parker Histology Section Head Skaggs Community Health Center Branson, Missouri _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Fri Jan 13 12:01:12 2006 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Jan 13 12:01:54 2006 Subject: [Histonet] Eosin too pink Message-ID: Try washing in tap water after eosin and before dehydrating. Experiment on times, I do 40 seconds. You'll probably also find better differentiation between various tissue elements. Fred >>> "Parker, Helayne" 01/13/06 11:42AM >>> Hi List, We are currently doing our H & E stain by hand and using the Richard-Allan 7211 Hematoxylin and their Eosin Y. Our pathologist is still complaining of slides being TOO PINK. I have cut the eosin step down to just 5 dips from 15 dips, and still too pink. The Eosin is very rich. This is our protocol: Hydrate to water like normal (thru Clear Rite 3, 100% and 95% Flex) Hematoxylin- 2 mins Rinsing in water until the water runs clear Clarifier II- 30 dips running water -30 dips Richard Allan Bluing - 30 dips running water - 30 dips 95% Flex - 10 dips Eosin-Y- 5 quick dips Thru 95%, 100% etc to xylene Any suggestions on a possible method change or a more mild Eosin. Complaint seems to solely "TOO PINK". I have done some experimenting w/ cutting the eosin w/ 80% Flex- and it does calm it down a bit but, the Hemo looks more purply then blue, almost periwinkle so not sure if that will be better or not- we still need to have him review the test slides. Any suggestions ? Helayne Parker Histology Section Head Skaggs Community Health Center Branson, Missouri _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BMolinari <@t> heart.thi.tmc.edu Fri Jan 13 11:57:38 2006 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Jan 13 12:03:03 2006 Subject: [Histonet] Eosin too pink Message-ID: Helayne, I have always stained my slides manually and I also use the same Richard Allen solutions. But my samples deparaffinize thru Xylenes - 100% (x3) to 95%. Then rinse in tap water and a rinse in DH2O. The following steps are all done for one minute: Hematoxylin Tap water rinse Clarifier (Richard Allan) Tap rinse Bluing (Richard Allan) Tap rinse 95% ETOH Eosin Y 100% ETOH (agitate the slides well to remove excess eosin) 100% ETOH 100% ETOH Xylenes x3 Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Parker, Helayne Sent: Friday, January 13, 2006 10:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Eosin too pink Hi List, We are currently doing our H & E stain by hand and using the Richard-Allan 7211 Hematoxylin and their Eosin Y. Our pathologist is still complaining of slides being TOO PINK. I have cut the eosin step down to just 5 dips from 15 dips, and still too pink. The Eosin is very rich. This is our protocol: Hydrate to water like normal (thru Clear Rite 3, 100% and 95% Flex) Hematoxylin- 2 mins Rinsing in water until the water runs clear Clarifier II- 30 dips running water -30 dips Richard Allan Bluing - 30 dips running water - 30 dips 95% Flex - 10 dips Eosin-Y- 5 quick dips Thru 95%, 100% etc to xylene Any suggestions on a possible method change or a more mild Eosin. Complaint seems to solely "TOO PINK". I have done some experimenting w/ cutting the eosin w/ 80% Flex- and it does calm it down a bit but, the Hemo looks more purply then blue, almost periwinkle so not sure if that will be better or not- we still need to have him review the test slides. Any suggestions ? Helayne Parker Histology Section Head Skaggs Community Health Center Branson, Missouri _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mtitford <@t> aol.com Fri Jan 13 12:05:54 2006 From: mtitford <@t> aol.com (mtitford@aol.com) Date: Fri Jan 13 12:06:04 2006 Subject: [Histonet] Dr Marshall & the English Language Message-ID: <8C7E68321AA83E7-D8C-A78@mblk-r18.sysops.aol.com> Dr Terry Marshall sayeth about H & E's that "he cannot have it too pink". Does he mean that he does not want it too pink, or that he likes it pink?! In any event, how can you readily detect eosinophilia when the eosin is too heavy? Mike Titford Pathology USA Mobile AL USA From dlcowie <@t> prodigy.net Fri Jan 13 12:16:29 2006 From: dlcowie <@t> prodigy.net (Dawn Cowie) Date: Fri Jan 13 12:16:39 2006 Subject: [Histonet] EGFR antibody Message-ID: <20060113181630.60627.qmail@web81005.mail.mud.yahoo.com> hello histonetters, I have a question. Which EGFR antibody is everyone using? Do you purchase Ventana's FDA approved antibody? Do you purchase Zymed's? etc. Also, why did you purchase the one over the others available? All responses to this will be helpful. Thanks, Dawn Cowie Histo Supv Pensacola Pathologists From jgemmanual <@t> wlgore.com Fri Jan 13 12:26:03 2006 From: jgemmanual <@t> wlgore.com (Jeannie G Emmanuel) Date: Fri Jan 13 12:26:19 2006 Subject: [Histonet] Dr Marshall & the English Language In-Reply-To: <8C7E68321AA83E7-D8C-A78@mblk-r18.sysops.aol.com> Message-ID: It means it will never be too pink for him. mtitford@aol.com Sent by: histonet-bounces@lists.utsouthwestern.edu 01/13/2006 11:05 AM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] Dr Marshall & the English Language Dr Terry Marshall sayeth about H & E's that "he cannot have it too pink". Does he mean that he does not want it too pink, or that he likes it pink?! In any event, how can you readily detect eosinophilia when the eosin is too heavy? Mike Titford Pathology USA Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sharon.osborn <@t> dnax.org Fri Jan 13 12:26:32 2006 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Jan 13 12:27:17 2006 Subject: [Histonet] H&E stainng Message-ID: <29B25753F6B1D51196110002A589D444032C497A@PALMSG30.us.schp.com> Helayne, Increase your time in the hematoxylin rather than trying to cut the eosin. We routinely do 6-8 minutes in hematoxylin. Most hematoxylins can stain for up to 15 minutes depending upon the strength of the hematoxylin and what the pathologist wants. This is the first step to try. If you were using longer times in the hematoxylin and had this problem, I would be concerned about your tap water washing out the hematoxylin. In that situation, it would depend upon the treatment of the tap water (large chlorine amounts, etc) that creates the difficulty(I encountered this problem in a hospital lab that had their own well water and self treated it). Again, try increasing your time in the hematoxylin and see if that solves your problem. By the way, you live in one of my favorite places, especially since I have several generations of family there! sharon osborn DNAX SP BioPharma Palo Alto, Ca Date: Fri, 13 Jan 2006 10:42:11 -0600 From: "Parker, Helayne" Subject: [Histonet] Eosin too pink To: Message-ID: <2FABC6145388F24EBA8CBD6EC165BCCB02AFB9E5@mail1-schc.skaggs.net> Content-Type: text/plain; charset="us-ascii" Hi List, We are currently doing our H & E stain by hand and using the Richard-Allan 7211 Hematoxylin and their Eosin Y. Our pathologist is still complaining of slides being TOO PINK. I have cut the eosin step down to just 5 dips from 15 dips, and still too pink. The Eosin is very rich. This is our protocol: Hydrate to water like normal (thru Clear Rite 3, 100% and 95% Flex) Hematoxylin- 2 mins Rinsing in water until the water runs clear Clarifier II- 30 dips running water -30 dips Richard Allan Bluing - 30 dips running water - 30 dips 95% Flex - 10 dips Eosin-Y- 5 quick dips Thru 95%, 100% etc to xylene Any suggestions on a possible method change or a more mild Eosin. Complaint seems to solely "TOO PINK". I have done some experimenting w/ cutting the eosin w/ 80% Flex- and it does calm it down a bit but, the Hemo looks more purply then blue, almost periwinkle so not sure if that will be better or not- we still need to have him review the test slides. Any suggestions ? Helayne Parker Histology Section Head Skaggs Community Health Center Branson, Missouri ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From PMonfils <@t> Lifespan.org Fri Jan 13 14:04:23 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Jan 13 14:04:33 2006 Subject: [Histonet] Eosin too pink Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717642@lsexch.lsmaster.lifespan.org> You can try rinsing the slides in tap water after staining, as someone already suggested. However , the pH of tap water differs considerably from one locality to another. If your tap water is slightly acidic it won't differentiate eosin at all. If it is very slightly basic (pH 7.1-7.2) you may be able to use it to remove excess eosin at a more or less controlled rate, and will have to experiment to determine the optimum time of exposure. But if your tap water is more strongly basic it may remove eosin so quickly that the differentiation is difficult to control, and uneven differentiation is likely to occur. I would feel safer using 95% ethanol to partially extract eosin from overstained tissues. Eosin dissolves out of tissue fairly quickly in 95% ethanol, but very slowly in 100% ethanol. In any such differentiation procedure the rack of slides should be frequently agitated in the solvent to ensure evenness of extraction throughout the section. That having been said, if I were in your position I wouldn't think in terms of correcting the overstaining, but preventing it. An extremely short staining time is likely to cause uneven staining (remember, the eosin has to extract the previous solvent, whether water or alcohol, from the tissue before it can get into the tissue and stain it). I would try diluting the eosin, using whatever solvent it is made with, until the desired intensity of staining can be achieved in a normal time period. From LuckG <@t> empirehealth.org Fri Jan 13 14:20:11 2006 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Jan 13 14:20:23 2006 Subject: [Histonet] Eosin too pink Message-ID: We use an 80% alcohol station immediately after the Eosin to accomplish a similar result. It's easy and consistent. -----Original Message----- From: Monfils, Paul [mailto:PMonfils@Lifespan.org] Sent: Friday, January 13, 2006 12:04 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Eosin too pink You can try rinsing the slides in tap water after staining, as someone already suggested. However , the pH of tap water differs considerably from one locality to another. If your tap water is slightly acidic it won't differentiate eosin at all. If it is very slightly basic (pH 7.1-7.2) you may be able to use it to remove excess eosin at a more or less controlled rate, and will have to experiment to determine the optimum time of exposure. But if your tap water is more strongly basic it may remove eosin so quickly that the differentiation is difficult to control, and uneven differentiation is likely to occur. I would feel safer using 95% ethanol to partially extract eosin from overstained tissues. Eosin dissolves out of tissue fairly quickly in 95% ethanol, but very slowly in 100% ethanol. In any such differentiation procedure the rack of slides should be frequently agitated in the solvent to ensure evenness of extraction throughout the section. That having been said, if I were in your position I wouldn't think in terms of correcting the overstaining, but preventing it. An extremely short staining time is likely to cause uneven staining (remember, the eosin has to extract the previous solvent, whether water or alcohol, from the tissue before it can get into the tissue and stain it). I would try diluting the eosin, using whatever solvent it is made with, until the desired intensity of staining can be achieved in a normal time period. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From llewllew <@t> shaw.ca Fri Jan 13 14:25:56 2006 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Jan 13 14:28:03 2006 Subject: [Histonet] Eosin too pink References: <09C945920A6B654199F7A58A1D7D1FDE01717642@lsexch.lsmaster.lifespan.org> Message-ID: <000501c6187f$8f3423f0$690a4246@yourlk4rlmsu> There have been many suggestions made so far, all valid. Here's another one. Try making a 0.1% solution of eosin Y in 95% ethanol, stain for 15 seconds with 2 seconds agitation at the beginning, then rinse thoroughly with gentle agitation in 95% ethanol for about 10 seconds just to remove the excess eosin solution (i.e. use it progressively). If still too pink, dilute in half and repeat. Do this until you have the required density. Cutting the staining time to 5 dips from 15 dips is pretty drastic (what kind of dips - how long is each dip?). Very short staining times are not usually the best, as has been said, so I suggest you keep the time standard and adjust the concentration. The best way to solve this kind of problem is always to run a systematic series of trials using sections of your common tissues with different concentrations of dye and times of application. Bryan Llewellyn From gcallis <@t> montana.edu Fri Jan 13 14:57:01 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Jan 13 14:57:14 2006 Subject: [Histonet] RE: Eosin too pink In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE01717642@lsexch.lsmaster.l ifespan.org> References: <09C945920A6B654199F7A58A1D7D1FDE01717642@lsexch.lsmaster.lifespan.org> Message-ID: <6.0.0.22.1.20060113134618.01b40288@gemini.msu.montana.edu> Paul made good suggestions here. Remember that carryover of eosin into the first 95% alcohol rinse can become excessive, so rotating or changing the 95% alcohol stations should be done more often (we sometimes do this daily with large volume staining sessions) to ensure proper differentiation or removal of excess eosin per his suggestion and as it was taught to me milleniums ago). If there is excessive carryover of eosin in the 95%, staining may continue with this dye - resulting is overly red tissues. If there is any pink or red tinge in the 100% alcohol or last station before clearing not only is dye being moved up the dehydration chain of events but also water (see Richard Allan website , go to their product Staining Guide.) At 01:04 PM 1/13/2006, you wrote: > You can try rinsing the slides in tap water after staining, as >someone already suggested. However , the pH of tap water differs >considerably from one locality to another. If your tap water is slightly >acidic it won't differentiate eosin at all. If it is very slightly basic (pH >7.1-7.2) you may be able to use it to remove excess eosin at a more or less >controlled rate, and will have to experiment to determine the optimum time >of exposure. But if your tap water is more strongly basic it may remove >eosin so quickly that the differentiation is difficult to control, and >uneven differentiation is likely to occur. I would feel safer using 95% >ethanol to partially extract eosin from overstained tissues. Eosin >dissolves out of tissue fairly quickly in 95% ethanol, but very slowly in >100% ethanol. In any such differentiation procedure the rack of slides >should be frequently agitated in the solvent to ensure evenness of >extraction throughout the section. > > That having been said, if I were in your position I wouldn't think >in terms of correcting the overstaining, but preventing it. An extremely >short staining time is likely to cause uneven staining (remember, the eosin >has to extract the previous solvent, whether water or alcohol, from the >tissue before it can get into the tissue and stain it). I would try >diluting the eosin, using whatever solvent it is made with, until the >desired intensity of staining can be achieved in a normal time period. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From PKamalavenkatesh <@t> wockhardtin.com Fri Jan 13 21:23:51 2006 From: PKamalavenkatesh <@t> wockhardtin.com (PKamalavenkatesh@wockhardtin.com) Date: Fri Jan 13 21:23:52 2006 Subject: [Histonet] cell proliferation markers Message-ID: Dear Histonetters, I am in need of details of few research papers involving methodology for the detection of cell proliferation markers such as PCNA and Ki 67 in paraffin sections. It would be greatly appreciated if anybody let me know the details. thanks kamalavenkatesh Please Visit our New Corporate Web Site www.wockhardt.com --------------------- Disclaimer ------------------------------------------------------------------------------ Information transmitted by this E-MAIL is proprietary to Wockhardt Ltd. and/ or its Group Companies and/or,its Customers and is intended for use only by the individual or entity to which it is addressed, and may contain information that is privileged, confidential or exempt from disclosure under applicable law. If you are not the intended recipient or it appears that this mail has been forwarded to you without proper authority, you are notified that any use or dissemination of this information in any manner is strictly prohibited. In such cases, please delete this mail from your records. --------------------------------------------------------------------------------------------------------------- From marktarango <@t> earthlink.net Sat Jan 14 20:03:56 2006 From: marktarango <@t> earthlink.net (Mark Adam Tarango) Date: Sat Jan 14 20:04:04 2006 Subject: [Histonet] RE: Eosin too pink Message-ID: <9221640.1137290636530.JavaMail.root@elwamui-hound.atl.sa.earthlink.net> So is he complaining that the eosin is too strong or just too pink? It seems to me like the more water you have in the alcohol after Eosin or if you use a water rinse after eosin it gets pinker....it's more orange when you have a higher percentage of alcohol in the alcohol after eosin(just seems that way to me anyway). If it's just too strong, dilute it down with alcohol. I hardly get any specimens, so I don't like changing the Eosin often, i just keep adding alcohol to it when it gets low and crusted around the edges..... still seems to work fine.... Mark T. > Sent: Friday, January 13, 2006 10:42 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Eosin too pink > > Hi List, > We are currently doing our H & E stain by hand and using the > Richard-Allan 7211 Hematoxylin and their Eosin Y. Our pathologist is > still complaining of slides being TOO PINK. I have cut the eosin step > down to just 5 dips from 15 dips, and still too pink. The Eosin is very > rich. > > This is our protocol: > Hydrate to water like normal (thru Clear Rite 3, 100% and 95% Flex) > Hematoxylin- 2 mins > Rinsing in water until the water runs clear > Clarifier II- 30 dips > running water -30 dips > Richard Allan Bluing - 30 dips > running water - 30 dips > 95% Flex - 10 dips > Eosin-Y- 5 quick dips > Thru 95%, 100% etc to xylene > > Any suggestions on a possible method change or a more mild Eosin. > Complaint seems to solely "TOO PINK". I have done some experimenting w/ > cutting the eosin w/ 80% Flex- and it does calm it down a bit but, the > Hemo looks more purply then blue, almost periwinkle so not sure if that > will be better or not- we still need to have him review the test slides. > Any suggestions ? > > Helayne Parker > Histology Section Head > Skaggs Community Health Center > Branson, Missouri From Barry.R.Rittman <@t> uth.tmc.edu Sun Jan 15 12:28:11 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Sun Jan 15 12:28:14 2006 Subject: [Histonet] RE: Eosin too pink Message-ID: If I were in your shoes I would try a weaker solution of esoin and if necessary prolong the times. I am also a believer in timing the stages of staining, as one perons two dips equals another persons three dips and so on. One would think that considering the amount of time that we all have been doing H and Es, that there should be a published standard image that we all could agree upon. This ain't gonna happen as the amount of staining with eosin that is regarded as desirable is generally a matter of personal taste. When stainig for our pathologist many moons ago, we stained deeply with eosin, when staning for the researcher the eosin was extermely weak. I am not convinced that either of them had a good grasp of the finer points of staining but they were the customers. I think that in talking about eosin staining, it should be borne in mind that the type of staining with aqueous and with alcoholic eosin solutions can differ considerably. Here in the states people have generally used alcoholic solutions of eosin. This gives a more rapid stain but in my opinion does not give the range of hues that can be obtained with aqueous solutions. I am used to using a solution of an aqueous solution, a mixture of eosin Y and eosin B (4 part to 1 part). Staining with this for 5 minutes and then carrying out most of the differentiation in tap water followed by a rapid dehydration. This used for all tissue except bone where a much more dilute solution for longer times is best. I believe that this provides a range of hues to the eosin staining that allows significantly better differentiation of structures. Heavy eosin staining tends to lose this differntiation. The only time that I used to stain more heavily with esoin was for photographing sections, and that is not necessary anymore due to the improved imaging techniques and manipulations that are easy to carry out. I think that as opinions are bound to differ, what we need is to compare what individuals regard as a good H and Es. How about sending in photos of what you regard as "good H and Es" to the Histonet picture site? I suggest that there be a limited number of tissues so that we do not end up with a logistic nightmare. How about liver, small intestine, uterus, skeletal muscle and skin? I think that for bone this might be best via the Hard Tissue Communique. Just my opinion. Barry ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Mark Adam Tarango Sent: Sat 1/14/2006 8:03 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Eosin too pink So is he complaining that the eosin is too strong or just too pink? It seems to me like the more water you have in the alcohol after Eosin or if you use a water rinse after eosin it gets pinker....it's more orange when you have a higher percentage of alcohol in the alcohol after eosin(just seems that way to me anyway). If it's just too strong, dilute it down with alcohol. I hardly get any specimens, so I don't like changing the Eosin often, i just keep adding alcohol to it when it gets low and crusted around the edges..... still seems to work fine.... Mark T. > Sent: Friday, January 13, 2006 10:42 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Eosin too pink > > Hi List, > We are currently doing our H & E stain by hand and using the > Richard-Allan 7211 Hematoxylin and their Eosin Y. Our pathologist is > still complaining of slides being TOO PINK. I have cut the eosin step > down to just 5 dips from 15 dips, and still too pink. The Eosin is very > rich. > > This is our protocol: > Hydrate to water like normal (thru Clear Rite 3, 100% and 95% Flex) > Hematoxylin- 2 mins > Rinsing in water until the water runs clear > Clarifier II- 30 dips > running water -30 dips > Richard Allan Bluing - 30 dips > running water - 30 dips > 95% Flex - 10 dips > Eosin-Y- 5 quick dips > Thru 95%, 100% etc to xylene > > Any suggestions on a possible method change or a more mild Eosin. > Complaint seems to solely "TOO PINK". I have done some experimenting w/ > cutting the eosin w/ 80% Flex- and it does calm it down a bit but, the > Hemo looks more purply then blue, almost periwinkle so not sure if that > will be better or not- we still need to have him review the test slides. > Any suggestions ? > > Helayne Parker > Histology Section Head > Skaggs Community Health Center > Branson, Missouri _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DKAEK <@t> coloplast.com Mon Jan 16 04:05:28 2006 From: DKAEK <@t> coloplast.com (Annette Ekblond) Date: Mon Jan 16 04:16:58 2006 Subject: [Histonet] marker of endothelial differentiation in vitro Message-ID: Dear subscribers -does anyone know of an endothelial marker specific to endothelial cells that have differentiated into tube-like structures (in vitro) - and not proliferating endothelial cultures?? Kind regards Annette CR Humlebaek Denmark From kbroomal <@t> NEMOURS.ORG Mon Jan 16 07:31:32 2006 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Mon Jan 16 07:31:57 2006 Subject: [Histonet] Desperately needing suggestions please! Message-ID: <6E41111281623B4B8A9AB8F9A7EA34378244C9@wlmmsx02.nemours.org> Does anyone know what I can use to get pararosaniline out of fabric? I was going to try acid alcohol, but haven't tried anything yet out of fear of making it worse. If anyone has suggestions, I and my brand new tan corduroy jack would much appreciate it! Kristen kbroomal@nemours.org From LJohns53 <@t> CNTUS.JNJ.COM Mon Jan 16 07:59:12 2006 From: LJohns53 <@t> CNTUS.JNJ.COM (Johns, Laura [CNTUS]) Date: Mon Jan 16 07:59:28 2006 Subject: [Histonet] Cell Pellets in Agar Message-ID: <70F83FE9F65318468A612768E7043F8903753FE6@cntusmaexs9.na.jnj.com> Does anyone have a protocol for paraffin embedding cell pellets? We've had problems with cell pellets staying together after fixation and during embedding, and I know that occasionally people use agar or agarose to keep the pellets compact. Is it better to use agar or agarose? Thanks, Laura M. Johns, Ph.D. Centocor, Inc. 145 King of Prussia Road (Mail Stop # R-4-2) Radnor, PA 19087 Phone: 610-240-8284 .... Fax: 610-240-8150.... Email: ljohns53@cntus.jnj.com > Confidentiality Notice: This message is intended for the individual(s) 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, he/she is > 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 by telephone and delete > the e-mail from your system immediately. Thank you. > > From sasa <@t> jovanovic.co.za Mon Jan 16 08:17:24 2006 From: sasa <@t> jovanovic.co.za (Sasa Jovanovic) Date: Mon Jan 16 08:18:00 2006 Subject: [Histonet] autofluorescence Message-ID: <000701c61aa7$9c07b810$640aa8c0@SinisaHome> Hi everyone Please help! In my project I am using immunohostochemistry to determine presence/absence of integrins in rat uterine tissue. I am trying to make it work for several months now with both alpha V beta 3 (immunoflorescence) and alpha 4 beta 1 integrin (using DAKO ARK kit) and use of monoclonal antibodies. My biggest problem so far is staining in the negative control (positive and negative control stain exactly the same) and auto florescence. I am following the Santa Cruz Protocol for immuno-fluorescence staining of frozen tissue. I snap freeze rat uterine tissue in liquid nitrogen, cut 6 um thin sections and adhere them on silane pre-treated slides and dry overnight...... then I fix them in cold acetone for 10 minutes and proceed with staining as follows: - wash slides in three changes of PBS (pH9) -incubate slides for 5-10 minutes in 0.1 H2O2 in PBS to quench endogenous peroxidase activity - wash slides in PBS 2x5minutes - incubate slides with 10% normal goat blocking serum in PBS for 20 min to suppress non - specific binding of Ig G - wash in PBS (3x5min) - incubate with primary antibody for 60 minutes - wash in 3 changes of PBS - incubate for 45 minutes with FITC conjugated secondary diluted in PBS with 1.5-3% normal blocking serum in dark chamber - wash in 3 changes of PBS - mount and coverslip directly from PBS with aqueous mounting medium Both primary (sc-7312)and secondary (sc-2010) antibody were purchased from SCBT I hope this is sufficient info to help you find some answers to my problem Thank you in advance Kind regards Sasha From Terry.Marshall <@t> rothgen.nhs.uk Mon Jan 16 08:27:15 2006 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Mon Jan 16 08:25:59 2006 Subject: [Histonet] Dr Marshall & the English Language Message-ID: Hmm. Can't have it too pink means one likes it pink. I did not realise in using this idiom that it is not universally understood. That's the problem with idioms. One does not ordinarily want to "detect eosinophilia". For that, the merest whiff of eosin is what is needed, and that is too my eye, hopeless for diagnostic work. Perhaps Mike means that there is not the nuance of tint if overstained. Of course. By overstating the case I was emphasising my preference to having a blue and red stain to a blue and itsi bitsi pink-in-places stain:_) Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: mtitford@aol.com [mailto:mtitford@aol.com] Sent: 13 January 2006 18:06 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dr Marshall & the English Language Dr Terry Marshall sayeth about H & E's that "he cannot have it too pink". Does he mean that he does not want it too pink, or that he likes it pink?! In any event, how can you readily detect eosinophilia when the eosin is too heavy? Mike Titford Pathology USA Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tyler-wellington <@t> northwestern.edu Mon Jan 16 08:49:49 2006 From: tyler-wellington <@t> northwestern.edu (Tyler W.) Date: Mon Jan 16 08:49:57 2006 Subject: [Histonet] DI Water Message-ID: <20060116144949.AB5D09E83C@merle.it.northwestern.edu> The DI water tap here in the lab isn't working. Does anyone know if its alright to use regular tap water in a flotation bath? From DonnaHansen <@t> chiwest.com Mon Jan 16 09:27:35 2006 From: DonnaHansen <@t> chiwest.com (Hansen, Donna (Ontario)) Date: Mon Jan 16 09:29:12 2006 Subject: [Histonet] DI Water Message-ID: <91FAC8D14C98974F839CA842BBC67D42136356@nwont2dc01.ontario-or.catholichealth.net> ________________________________ From: Hansen, Donna (Ontario) Sent: Mon 1/16/2006 8:21 AM To: Tyler W. Subject: RE: [Histonet] DI Water Good morning. We have always used tap water in the water bath. flotation bath, water bath, same thing? ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tyler W. Sent: Mon 1/16/2006 7:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] DI Water The DI water tap here in the lab isn't working. Does anyone know if its alright to use regular tap water in a flotation bath? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Mon Jan 16 09:31:14 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Jan 16 09:31:26 2006 Subject: [Histonet] Cell Pellets in Agar Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717643@lsexch.lsmaster.lifespan.org> You can use a number of soluble, coagulable proteins to bind cell pellets together, including agar, agarose, gelatin, albumin, serum, etc. I use Mayer's albumin, but the technique is pretty much the same regardless of which substance is used. After fixing the cells in suspension, I spin them down and remove the fixative. Then wash a couple of times in buffer by resuspending and spinning down. After the last wash I resuspend in buffer once more, add 2 drops of Mayer's albumin per ml of suspension, mix thoroughly by vortexing a few seconds, allow to stand for a couple of minutes, then spin down once more and remove the supernatant. I then begin dehydration of the pellet in the centrifuge tube, usually beginning with 50% ethanol, then 70%, 95%, and absolute ethanol. Pipette the initial alcohol gently down the side of the tube to ensure that the pellet is not disturbed. Once you are in the stronger alcohols the thin coating of albumin on the cells has coagulated, binding them firmly together. At some point in the dehydration series the pellet, due to contraction during dehydration, usually detaches from the tube intact. If it doesn't actually detach, it can be detached either by drawing up alcohol into a pipette and squirting it onto the pellet, or if necessary, by the slightest touch of a slender probe. I then either wrap the pellet in lens paper and place it in a cassette on the tissue processor, starting with the final absolute alcohol station, or, I finish the processing by hand, in the centrifuge tube. If you are going to do the latter, make sure your tube is resistant to the clearing agent. Polypropylene tubes work best for me. Polystyrene tubes are more transparent, and are fine if you are only going to use alcohol in the tube, but many clearing agents dissolve polystyrene. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Johns, Laura [CNTUS] > Sent: Monday, January 16, 2006 5:59 AM > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] Cell Pellets in Agar > > Does anyone have a protocol for paraffin embedding cell pellets? We've > had > problems with cell pellets staying together after fixation and during > embedding, and I know that occasionally people use agar or agarose to keep > the pellets compact. Is it better to use agar or agarose? > > Thanks, > > Laura M. Johns, Ph.D. > Centocor, Inc. > 145 King of Prussia Road (Mail Stop # R-4-2) > Radnor, PA 19087 > Phone: 610-240-8284 .... Fax: 610-240-8150.... Email: > ljohns53@cntus.jnj.com > > > Confidentiality Notice: This message is intended for the individual(s) > 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, he/she is > > 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 by telephone and delete > > the e-mail from your system immediately. Thank you. > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From PMonfils <@t> Lifespan.org Mon Jan 16 09:57:35 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Jan 16 09:57:50 2006 Subject: [Histonet] Desperately needing suggestions please! Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717644@lsexch.lsmaster.lifespan.org> For basic dyes like pararosaniline or hematoxylin, acid alcohol would probably be your best bet. I keep a little bottle of acid alcohol (fairly strong - about 4% hydrochloric acid in 70% ethanol) in the cabinet above the washer at home, to pretreat such stains. Acid dyes like eosin or fast green will usually wash out pretty easily without pretreatment because of the basic pH of the tap water and the detergent. But those same basic properties will set basic dyes rather than remove them. I just saturate the stain with a little acid alcohol, work it in a bit, then blot it firmly between paper towels (under and on top of the fabric) (pound on it a bit actually), repeating if necessary, and then when I have sufficiently removed the stain, drop the garment into the already running washer. Sounds like your concern may be a non-washable garment, in which case the saturate and blot technique alone will HOPEFULLY be sufficient. In that case I think I would finish up with one last saturate and blot using either plain 70% alcohol or distilled water, to remove excess acid from the fabric. I haven't seen a case where acid alcohol had any direct effect on fabric color, but you might want to try it on an inconspicuous part of the garment first, just to be sure. My wife once got some black grease on a white blouse that had little green leaves and pink and blue flowers. I brought the blouse in to work and treated it with xylene, which didn't completely remove the grease. They I tried chloroform, which quickly removed the grease, and the blue flowers. From hfedor <@t> jhmi.edu Mon Jan 16 09:59:07 2006 From: hfedor <@t> jhmi.edu (Helen Fedor) Date: Mon Jan 16 09:59:27 2006 Subject: [Histonet] Cell Pellets in Agar Message-ID: Paul, Here is the protocol that we use in our Lab. 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. >>> "Monfils, Paul" 01/16/06 10:31 AM >>> You can use a number of soluble, coagulable proteins to bind cell pellets together, including agar, agarose, gelatin, albumin, serum, etc. I use Mayer's albumin, but the technique is pretty much the same regardless of which substance is used. After fixing the cells in suspension, I spin them down and remove the fixative. Then wash a couple of times in buffer by resuspending and spinning down. After the last wash I resuspend in buffer once more, add 2 drops of Mayer's albumin per ml of suspension, mix thoroughly by vortexing a few seconds, allow to stand for a couple of minutes, then spin down once more and remove the supernatant. I then begin dehydration of the pellet in the centrifuge tube, usually beginning with 50% ethanol, then 70%, 95%, and absolute ethanol. Pipette the initial alcohol gently down the side of the tube to ensure that the pellet is not disturbed. Once you are in the stronger alcohols the thin coating of albumin on the cells has coagulated, binding them firmly together. At some point in the dehydration series the pellet, due to contraction during dehydration, usually detaches from the tube intact. If it doesn't actually detach, it can be detached either by drawing up alcohol into a pipette and squirting it onto the pellet, or if necessary, by the slightest touch of a slender probe. I then either wrap the pellet in lens paper and place it in a cassette on the tissue processor, starting with the final absolute alcohol station, or, I finish the processing by hand, in the centrifuge tube. If you are going to do the latter, make sure your tube is resistant to the clearing agent. Polypropylene tubes work best for me. Polystyrene tubes are more transparent, and are fine if you are only going to use alcohol in the tube, but many clearing agents dissolve polystyrene. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Johns, Laura [CNTUS] > Sent: Monday, January 16, 2006 5:59 AM > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] Cell Pellets in Agar > > Does anyone have a protocol for paraffin embedding cell pellets? We've > had > problems with cell pellets staying together after fixation and during > embedding, and I know that occasionally people use agar or agarose to keep > the pellets compact. Is it better to use agar or agarose? > > Thanks, > > Laura M. Johns, Ph.D. > Centocor, Inc. > 145 King of Prussia Road (Mail Stop # R-4-2) > Radnor, PA 19087 > Phone: 610-240-8284 .... Fax: 610-240-8150.... Email: > ljohns53@cntus.jnj.com > > > Confidentiality Notice: This message is intended for the individual(s) > 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, he/she is > > 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 by telephone and delete > > the e-mail from your system immediately. Thank you. > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sharon.osborn <@t> dnax.org Mon Jan 16 12:08:31 2006 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Mon Jan 16 12:10:14 2006 Subject: [Histonet] IHC slides losing tissue Message-ID: <29B25753F6B1D51196110002A589D444032C4989@PALMSG30.us.schp.com> Histonetters, One of our techs cut ovary at 5u paraffin sections placed on charged slides. These were air dried but not deparaffinized in the oven. The researcher deparaffinized through zylene then placed in the pressure cooker at 60 for antigen retrieval. The tissue came off. Our first remedy is to oven deparaffinize the slides then do the retrieval process. If that is not successful, what is the solution(s) for this problem. Our senior IHC tech suggested gelatin subbing on the slides but did not think that would solve the problem. Would there be too much background with the gelatin should it be successful? What is your experience in what works best. Much thanks... Sharon Osborn DNAX, SP BioPharma Palo Alto, CA 650.946.6539 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From BlazekL <@t> childrensdayton.org Mon Jan 16 13:22:23 2006 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Mon Jan 16 13:25:01 2006 Subject: [Histonet] IHC slides losing tissue Message-ID: The first thing I would check is to see if there was any additive to the waterbath. By putting an adhesive in the water bath the sections are almost guaranteed to fall off. Next I would be sure the slides were well dried. I put mine in the oven over night at 56 degrees C. Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Osborn, Sharon" 01/16/2006 1:08:31 PM >>> Histonetters, One of our techs cut ovary at 5u paraffin sections placed on charged slides. These were air dried but not deparaffinized in the oven. The researcher deparaffinized through zylene then placed in the pressure cooker at 60 for antigen retrieval. The tissue came off. Our first remedy is to oven deparaffinize the slides then do the retrieval process. If that is not successful, what is the solution(s) for this problem. Our senior IHC tech suggested gelatin subbing on the slides but did not think that would solve the problem. Would there be too much background with the gelatin should it be successful? What is your experience in what works best. Much thanks... Sharon Osborn DNAX, SP BioPharma Palo Alto, CA 650.946.6539 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon Jan 16 13:44:09 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Jan 16 13:44:20 2006 Subject: [Histonet] IHC slides losing tissue In-Reply-To: <29B25753F6B1D51196110002A589D444032C4989@PALMSG30.us.schp.com> Message-ID: <20060116194409.10155.qmail@web61220.mail.yahoo.com> Sharon: If you just air dried the sections on the slides before doing the antigen retrieval in a pressure cooker (where the temperature is >100?C) the sections were doomed to fall. Just to be clear you don't deparafinize in the oven, what you do in te oven is to melt the paraffin and asure that the sections get in close contact with the glass that, is (+) charged will retain the sections during the IHC procedure. To succeed in this step, you have to first make sure that there is no water left between the section and the slide. Thenyou will have to put your slides in a oven that could be at 60?C and leave them there for at least ?hour. Take them out and let them cool to room temperature. After that you can either store them or use them for any staining procedure. Sections as thin as the ones you describe (5?m) should survive. You don't really want to ad gelatin neither on the slide or in the water bath. In both instances you will end with background because you have to remember that gelatins are proteins. As a matter of fact the water bath for IHC section should be filled with distilled water alone. "Osborn, Sharon" wrote: Histonetters, One of our techs cut ovary at 5u paraffin sections placed on charged slides. These were air dried but not deparaffinized in the oven. The researcher deparaffinized through zylene then placed in the pressure cooker at 60 for antigen retrieval. The tissue came off. Our first remedy is to oven deparaffinize the slides then do the retrieval process. If that is not successful, what is the solution(s) for this problem. Our senior IHC tech suggested gelatin subbing on the slides but did not think that would solve the problem. Would there be too much background with the gelatin should it be successful? What is your experience in what works best. Much thanks... Sharon Osborn DNAX, SP BioPharma Palo Alto, CA 650.946.6539 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos ? Showcase holiday pictures in hardcover Photo Books. You design it and we?ll bind it! From beth <@t> hsrl.org Wed Jan 11 16:34:40 2006 From: beth <@t> hsrl.org (Beth) Date: Mon Jan 16 14:41:54 2006 Subject: [Histonet] GMS on frozen sections? Message-ID: <000001c616ff$36401a00$0200a8c0@HSRLMAIN> Dear Netters, Does anyone have a protocol for GMS staining of OCT embedded tissues? Any help would be appreciated. Sincerely, Beth Poole HSRL, Inc.- A GLP Compliant Contract Laboratory 5930 Main Street Mount Jackson, Virginia 22842 540.477.4440 Fax: 540.477.4448 beth@hsrl.org www.hsrl.org From YINM <@t> ccf.org Thu Jan 12 09:39:43 2006 From: YINM <@t> ccf.org (Yin, Mei) Date: Mon Jan 16 14:41:55 2006 Subject: [Histonet] Protocol for GMA Message-ID: <2B24BD7D5BCF5342B5848D06C995B433CB0AAE@CCHSCLEXMB56.cc.ad.cchs.net> Dear Mr. Ms. This is Mei Yin and I work in Cleveland Clinic Foundation. I am going to do acetone fixation/GMA embedding for light microscopy. However, I never have done this before. Therefore, I want to ask you if you can help me and show me the protocol I will be very appreciate. Mei Yin My lab phone number is 216-444-8560 ------------------------------------------------------------------------------ Confidentiality Note: This message is intended for use only by the individual or entity to which it is addressed and may contain information that is privileged, confidential, and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please contact the sender immediately and destroy the material in its entirety, whether electronic or hard copy. Thank you. ------------ Visit us online at our award-winning http://www.clevelandclinic.org for a complete listing of Cleveland Clinic services, staff and locations from one of the country's leading hospitals. ============================================================================== From BJDewe <@t> aol.com Mon Jan 16 15:42:51 2006 From: BJDewe <@t> aol.com (BJDewe@aol.com) Date: Mon Jan 16 15:43:06 2006 Subject: [Histonet] IHC slides losing tissue Message-ID: In a message dated 1/16/2006 11:45:34 AM Pacific Standard Time, rjbuesa@yahoo.com writes: Just to be clear you don't deparafinize in the oven, what you do in te oven is to melt the paraffin and asure that the sections get in close contact with the glass that, is (+) charged will retain the sections during the IHC procedure. This is an interesting observation. I have always done IHC on sections that were airdried at RT overnight and had good results but many times my IHC protocols are lengthy and most recently, after the entire IHC process, my sections fell off in the dehydration to go to xylene to coverslip. Very frustrating let alone a waste of time and money... I will try this new method and hope for better results ;-) Thanks!! Loralei Dewe UC Davis VM: Surgical and Radiological Sciences From Jackie.O'Connor <@t> abbott.com Mon Jan 16 16:15:50 2006 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Mon Jan 16 16:16:20 2006 Subject: [Histonet] CD19 FFPE or zinc Message-ID: Has anyone ever found a CD19 antibody that works on human or mouse FFPE or zinc fixed PE? Gracias. Jackie O' From brian.chelack <@t> usask.ca Mon Jan 16 16:38:26 2006 From: brian.chelack <@t> usask.ca (Brian Chelack) Date: Mon Jan 16 16:39:11 2006 Subject: [Histonet] Re: cells in agar Message-ID: <002501c61aed$90ecd7d0$4f13e980@PDS11> I have had good success with a bit simpler method of creating a cell pellet that can be subsequently fixed, sectioned and immunostained. Melt a small volume of 2% low melting point agarose in normal saline. Hold at 37C. Re-suspend the cells of interest at about 1 10e6/ml and mix (using a Pasteur pipette) with an equal quantity of the melted agarose. Now to make real nice little agarose plugs I quickly add the still melted mixture into wells of a 96 well polyethylene plate (one of the fexible ones) and place it in the fridge for a few minutes. After the agarose/cell mixture is set, pop out the agarose plugs and place directly into 10% NBF. After fixation, the plugs can be transected and processed into paraffin blocks. If you don't have a polyethylene 96 well plate you can just drip the mixture onto a glass slide and peel off the drops after they have set. A bit low tech, but it works for me. Brian Chelack Prairie Diagnostic Services 2604-52 Campus Drive Saskatoon SK S7N 5B4 306-966-7241 From s03355926 <@t> yahoo.co.jp Tue Jan 17 01:30:24 2006 From: s03355926 <@t> yahoo.co.jp (yining zhsng) Date: Tue Jan 17 01:30:38 2006 Subject: [Histonet] how to distinguish liver sinusoidal endothelial cells from myofbroblast or hepatic stellate cells? Message-ID: <20060117073024.21962.qmail@web3208.mail.bbt.yahoo.co.jp> hi I am a graduate student working on hepatic stellate cells(HSC).A mesenchymal cells closes to myofibroblast on phenotype. I was required recently by a reviewer to mark out the location of liver sinusoidal endothelial cells (LSEC) on rat liver tissue to verify that a-SMA positive cells I showed are perisinusoidal cells.I tried to find out some markers for LSEC,such as CD31,CD34 and VWF,etc.But the comment for these markers are controversial.CD31 seems the best option but some papers mentioned that it is also expressed by myofibroblast & HSC.Does anyone give me any advices how can I demonstrate that HSCs,a-SMA positive cells are persinusoidal cells? Any information would be help and thanks for your help. yining Division of Gastroenterology and Hepatology, > University of Tsukuba > 1-1-1 Tennoudai, Tsukuba City, Japan 305-8575 -------------------------------------- GANBARE! NIPPON! Yahoo! JAPAN JOC OFFICIAL INTERNET PORTAL SITE PARTNER http://pr.mail.yahoo.co.jp/ganbare-nippon/ From wasielewski.reinhard.von <@t> mh-hannover.de Tue Jan 17 01:46:00 2006 From: wasielewski.reinhard.von <@t> mh-hannover.de (wasielewski.reinhard.von@mh-hannover.de) Date: Tue Jan 17 01:46:22 2006 Subject: [Histonet] Re: Cell pellets embedding in paraffin In-Reply-To: Message-ID: <43CCAEC8.2256.D26DFC2F@localhost> The answers given seem to be rather complicated. We just embedd the cell lines in 1% agarose and put it in the tissue processor - that's it. For details, see our paper: > Mengel M, Hebel K, Kreipe H, von Wasielewski R: Standardized on-slide control for quality assurance in the immunohistochemical assessment of therapeutic target molecules in breast cancer. Breast J 2005; 11(1): 34-40 It is really simple. With best regards Reinhard von Wasielewski PD Dr. med. Reinhard von Wasielewski From billions <@t> public1.sz.js.cn Tue Jan 17 02:11:37 2006 From: billions <@t> public1.sz.js.cn (Sinoera Tech) Date: Tue Jan 17 02:11:57 2006 Subject: [Histonet] Re: Alcian Yellow & Alcian Blue 8GX References: <43CCAEC8.2256.D26DFC2F@localhost> Message-ID: <000401c61b3d$a5f88340$e401a8c0@sinoera63d3596> Dear Sirs, We are supplying Alcian Yellow, Alcian Blue 8GX, Alcian Green. Please contact us for requirement. Kind Regards. - SUZHOU SINOERA CHEM CO., LTD. 125 Binhe Road Suzhou New & Hi-Tech District 215011 China Fax: +86 512 68258994 Tel: +86 512 68246939 http://www.sinoeratech.com http://www.sinoerachem.com From carl.hobbs <@t> kcl.ac.uk Tue Jan 17 03:00:13 2006 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Tue Jan 17 03:00:45 2006 Subject: [Histonet] paraformaldehyde crystals v Prills Message-ID: <001701c61b44$6d730b60$112b5c9f@Carlos> Many of our Groups use paraformaldehyde crystals ( Sigma P6148) to make up their solution of formalin. Works very well; 4g + 1tab PBS + 6microlitre 10MNaOH.....stir overnight. Dissolved completely:pH7.2 I am trying to get away from using this paraformaldehyde: it is very light and prone to spread around the fume hood when it's weighed out. So, I bought PRILLS from Sigma:Problem is, they just do not dissolve completely, unless I preboil the water.( heating to 60C is not hot enough to dissolve the Prills. I have tried all combinations of the formulation above( eg: no NaOH, dissolving PBS tablet before adding paraformaldehyde). I want the procedure with the least risk; boiling may be a higher risk than the spread of powder. NB: I cannot turn down/off the fume hoods, for the weighing of the crystals( which behave more like a very fine powder), hence my desire to use the heavier Prills. Be grateful for any insights. Carl From lpwenk <@t> sbcglobal.net Tue Jan 17 03:21:51 2006 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Tue Jan 17 03:22:16 2006 Subject: [Histonet] Desperately needing suggestions please! In-Reply-To: <6E41111281623B4B8A9AB8F9A7EA34378244C9@wlmmsx02.nemours.org> Message-ID: Anatech has a lotion that removes Schiffs from hands, and can also remove it from slides. I don't know if it can remove it from clothes, nor do I know what it would do to colored fabric. Try contacting them 1-800-ANATECH. http://www.anatechltdusa.com/Catalog/catalog_hndclnrs.html 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 Kristen Broomall Sent: Monday, January 16, 2006 8:32 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Desperately needing suggestions please! Does anyone know what I can use to get pararosaniline out of fabric? I was going to try acid alcohol, but haven't tried anything yet out of fear of making it worse. If anyone has suggestions, I and my brand new tan corduroy jack would much appreciate it! Kristen kbroomal@nemours.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Tue Jan 17 05:22:41 2006 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Tue Jan 17 05:22:51 2006 Subject: [Histonet] Fixation time Message-ID: <003b01c61b58$54ac2e40$eeeea8c0@SERVER01> I have a question to all fixation-experts. Can anyone tell me exact pentration- and fixation times, or the mm/hour for 4-8% NBF. I know it depends on temperature, tissue-quality, but I have not found a table about this. In "Histologische Technik, H.C. Burck" stands, that NBF takes 16 hours for 4mm in liver. (1mm/h) In the Microwave-book, Kok and Boon, stands NBF penetrates fast and the linkages develope slowly. But was is fast and slowly? In daily work we let the tissue-blocks beetween 4-8 hours in NBF. Biopsies sometimes only 2 hours or less. Is the ratio 1mm/h really the rule of thumb for penetration and fixation? Thank you Gudrun Lang Histolog. Labor Akh Linz ?sterreich From sharon.willman <@t> bms.com Tue Jan 17 06:34:22 2006 From: sharon.willman <@t> bms.com (Sharon E Willman) Date: Tue Jan 17 06:34:37 2006 Subject: [Histonet] Dry Stains Shelf Life Message-ID: <43CCE44E.7090107@bms.com> Hi, What is the expiration time for dry stains and how often should you replenish? Many times they do not have expiration dates on them when they come in from the manufacturer. I would appreciate any information on this matter. Thanks, Sharon Willman From rjbuesa <@t> yahoo.com Tue Jan 17 06:54:27 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Jan 17 06:54:37 2006 Subject: [Histonet] Dry Stains Shelf Life In-Reply-To: <43CCE44E.7090107@bms.com> Message-ID: <20060117125427.89646.qmail@web61213.mail.yahoo.com> Hi Sharon: My personal experience is that good quality dry stains are very stable and can be used "to the end" without any fear. As an example I can tell you that in the 1960s I was still using dry stains manufactured by Merck (Darmstad, Germany) just after the "Great War", meaning the First World War. What I used to do in terms of inspections was to place a label on the bottles with dry stains stating: "no expiration date". Hope this will help. Ren? J. Sharon E Willman wrote: Hi, What is the expiration time for dry stains and how often should you replenish? Many times they do not have expiration dates on them when they come in from the manufacturer. I would appreciate any information on this matter. Thanks, Sharon Willman _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From cwscouten <@t> myneurolab.com Tue Jan 17 07:51:00 2006 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Tue Jan 17 07:52:29 2006 Subject: [Histonet] Fixation time Message-ID: <5784D843593D874C93E9BADCB87342AB916AD5@tpiserver03.Coretech-holdings.com> The answer to this was posted in a lengthy article on the histonet about 2 years ago. I saved it, it was very good, I would like to credit the author but forget the name. Take a bow if you are reading this. Meanwhile, you can search the archives but I will forward you an attachment of the article. Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Tuesday, January 17, 2006 5:23 AM To: Histonetliste (Histonetliste) Subject: [Histonet] Fixation time I have a question to all fixation-experts. Can anyone tell me exact pentration- and fixation times, or the mm/hour for 4-8% NBF. I know it depends on temperature, tissue-quality, but I have not found a table about this. In "Histologische Technik, H.C. Burck" stands, that NBF takes 16 hours for 4mm in liver. (1mm/h) In the Microwave-book, Kok and Boon, stands NBF penetrates fast and the linkages develope slowly. But was is fast and slowly? In daily work we let the tissue-blocks beetween 4-8 hours in NBF. Biopsies sometimes only 2 hours or less. Is the ratio 1mm/h really the rule of thumb for penetration and fixation? Thank you Gudrun Lang Histolog. Labor Akh Linz ?sterreich _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Tue Jan 17 09:15:39 2006 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Tue Jan 17 09:16:21 2006 Subject: [Histonet] Fixation time Message-ID: <5784D843593D874C93E9BADCB87342AB916AD8@tpiserver03.Coretech-holdings.com> Sorry, I sent the attachment, but it was the wrong one. I can't find the right one on my computer. Search the histonet archives. Search for reference to Cammermeyer, that should find just that paper. The answer to this was posted in a lengthy article on the histonet about 2 years ago. I saved it, it was very good, I would like to credit the author but forget the name. Take a bow if you are reading this. Meanwhile, you can search the archives but I will forward you an attachment of the article. Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Tuesday, January 17, 2006 5:23 AM To: Histonetliste (Histonetliste) Subject: [Histonet] Fixation time I have a question to all fixation-experts. Can anyone tell me exact pentration- and fixation times, or the mm/hour for 4-8% NBF. I know it depends on temperature, tissue-quality, but I have not found a table about this. In "Histologische Technik, H.C. Burck" stands, that NBF takes 16 hours for 4mm in liver. (1mm/h) In the Microwave-book, Kok and Boon, stands NBF penetrates fast and the linkages develope slowly. But was is fast and slowly? In daily work we let the tissue-blocks beetween 4-8 hours in NBF. Biopsies sometimes only 2 hours or less. Is the ratio 1mm/h really the rule of thumb for penetration and fixation? Thank you Gudrun Lang Histolog. Labor Akh Linz ?sterreich _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From awatanabe <@t> tgen.org Tue Jan 17 09:24:01 2006 From: awatanabe <@t> tgen.org (Aprill Watanabe) Date: Tue Jan 17 09:24:12 2006 Subject: [Histonet] Re: Cell Pellets in agar In-Reply-To: <20060116183955.A12612000746@mr1.tgen.org> Message-ID: This is the protocol that I use and it works very well. Preparation of Cell Culture Blocks as TMA Block Donors 1) Passage cell lines 1.5-2 days before intended harvest date to ensure cells are in log phase of the cell growth curve. 2) Use flasks that are 75-85% confluent. 3) If using adherent cells, detach the cells with trypsin while treating cells gently. 4) Collect detached cells, centrifuge for 10 mins at 500xg. 5) Wash once in 1X PBS and pellet again. Remove supernatant. Proceed to step #9. 6) If using suspension cells, pellet 50 ml of cell culture supernatant in 50 ml conical tubes at 500xg for 10 min. 7) Aspirate off supernatant. 8) Wash with 1X PBS and pellet again. Remove supernatant. 9) Suspend cell pellets in 10 ml of 10% Neutral Buffered Formalin 10) Incubate at 4OC for 2-3 hrs (for suspension cells), overnight on shaker at room temperature for adherent cells. 11) Make up a 0.8-1% agarose solution and store at 60OC. 12) Add 100 ul of agarose to labeled eppendorf tubes and allow to harden. 13) Pellet cells at 500xg for 10 min. 14) Aspirate off supernatant (Try and get as much as possible) 15) Add 1 drop of Eosin to each cell pellet in the 50 ml tubes. 16) Incubate at RT for 5 min. 17) Suspended cell pellet in 300 ml of warm agarose. a. Keep the agarose in the 60OC chamber. b. You only get about enough time to pipet them up and down 3-4 times. c. Use 1 ml tips with the ends cut off so they don?t get clogged. d. Be careful while suspending pellets as bubbles form quite easily. 18) Add agar-cell suspension to respective eppendorf with agar plug. a. Add a 10 ml pipet tip to plug to allow you to pull the plugs out. 19) Allow agar-cell suspension to harden for 15 minutes. 20) Add ~1 ml of 10% Neutral Buffered Formalin to each tube. 21) Incubate O/N at 4OC. Send for paraffin embedding the following morning using short protocol. On 1/16/06 11:39 AM, "histonet-request@lists.utsouthwestern.edu" wrote: > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. RE: RE: Eosin too pink (Rittman, Barry R) > 2. marker of endothelial differentiation in vitro (Annette Ekblond) > 3. Desperately needing suggestions please! (Kristen Broomall) > 4. Cell Pellets in Agar (Johns, Laura [CNTUS]) > 5. autofluorescence (Sasa Jovanovic) > 6. RE: Dr Marshall & the English Language > (Marshall Terry Dr, Consultant Histopathologist) > 7. DI Water (Tyler W.) > 8. RE: DI Water (Hansen, Donna (Ontario)) > 9. RE: Cell Pellets in Agar (Monfils, Paul) > 10. RE: Desperately needing suggestions please! (Monfils, Paul) > 11. RE: Cell Pellets in Agar (Helen Fedor) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sun, 15 Jan 2006 12:28:11 -0600 > From: "Rittman, Barry R" > Subject: RE: [Histonet] RE: Eosin too pink > To: "histonet" > Message-ID: > > Content-Type: text/plain; charset="iso-8859-1" > > If I were in your shoes I would try a weaker solution of esoin and if > necessary prolong the times. > I am also a believer in timing the stages of staining, as one perons two dips > equals another persons three dips and so on. > > One would think that considering the amount of time that we all have been > doing H and Es, that there should be a published standard image that we all > could agree upon. This ain't gonna happen as the amount of staining with > eosin that is regarded as desirable is generally a matter of personal taste. > When stainig for our pathologist many moons ago, we stained deeply with eosin, > when staning for the researcher the eosin was extermely weak. I am not > convinced that either of them had a good grasp of the finer points of staining > but they were the customers. > > I think that in talking about eosin staining, it should be borne in mind that > the type of staining with aqueous and with alcoholic eosin solutions can > differ considerably. > Here in the states people have generally used alcoholic solutions of eosin. > This gives a more rapid stain but in my opinion does not give the range of > hues that can be obtained with aqueous solutions. > I am used to using a solution of an aqueous solution, a mixture of eosin Y and > eosin B (4 part to 1 part). Staining with this for 5 minutes and then > carrying out most of the differentiation in tap water followed by a rapid > dehydration. This used for all tissue except bone where a much more dilute > solution for longer times is best. > I believe that this provides a range of hues to the eosin staining that allows > significantly better differentiation of structures. Heavy eosin staining tends > to lose this differntiation. > The only time that I used to stain more heavily with esoin was for > photographing sections, and that is not necessary anymore due to the improved > imaging techniques and manipulations that are easy to carry out. > I think that as opinions are bound to differ, what we need is to compare what > individuals regard as a good H and Es. > How about sending in photos of what you regard as "good H and Es" to the > Histonet picture site? > I suggest that there be a limited number of tissues so that we do not end up > with a logistic nightmare. > How about liver, small intestine, uterus, skeletal muscle and skin? > I think that for bone this might be best via the Hard Tissue Communique. > Just my opinion. > Barry > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Mark Adam Tarango > Sent: Sat 1/14/2006 8:03 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: Eosin too pink > > > > So is he complaining that the eosin is too strong or just too pink? It seems > to me like the more water you have in the alcohol after Eosin or if you use a > water rinse after eosin it gets pinker....it's more orange when you have a > higher percentage of alcohol in the alcohol after eosin(just seems that way to > me anyway). If it's just too strong, dilute it down with alcohol. I hardly > get any specimens, so I don't like changing the Eosin often, i just keep > adding alcohol to it when it gets low and crusted around the edges..... still > seems to work fine.... > > Mark T. > >> Sent: Friday, January 13, 2006 10:42 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Eosin too pink >> >> Hi List, >> We are currently doing our H & E stain by hand and using the >> Richard-Allan 7211 Hematoxylin and their Eosin Y. Our pathologist is >> still complaining of slides being TOO PINK. I have cut the eosin step >> down to just 5 dips from 15 dips, and still too pink. The Eosin is very >> rich. >> >> This is our protocol: >> Hydrate to water like normal (thru Clear Rite 3, 100% and 95% Flex) >> Hematoxylin- 2 mins >> Rinsing in water until the water runs clear >> Clarifier II- 30 dips >> running water -30 dips >> Richard Allan Bluing - 30 dips >> running water - 30 dips >> 95% Flex - 10 dips >> Eosin-Y- 5 quick dips >> Thru 95%, 100% etc to xylene >> >> Any suggestions on a possible method change or a more mild Eosin. >> Complaint seems to solely "TOO PINK". I have done some experimenting w/ >> cutting the eosin w/ 80% Flex- and it does calm it down a bit but, the >> Hemo looks more purply then blue, almost periwinkle so not sure if that >> will be better or not- we still need to have him review the test slides. >> Any suggestions ? >> >> Helayne Parker >> Histology Section Head >> Skaggs Community Health Center >> Branson, Missouri > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > ------------------------------ > > Message: 2 > Date: Mon, 16 Jan 2006 11:05:28 +0100 > From: Annette Ekblond > Subject: [Histonet] marker of endothelial differentiation in vitro > To: histonet@lists.utsouthwestern.edu > Message-ID: > > > Content-Type: text/plain; charset=US-ASCII > > > Dear subscribers > > -does anyone know of an endothelial marker specific to endothelial cells that > have differentiated into tube-like > structures (in vitro) - and not proliferating endothelial cultures?? > > Kind regards > > Annette > CR > Humlebaek > Denmark > > > > > ------------------------------ > > Message: 3 > Date: Mon, 16 Jan 2006 08:31:32 -0500 > From: "Kristen Broomall" > Subject: [Histonet] Desperately needing suggestions please! > To: Histonet@lists.utsouthwestern.edu > Message-ID: > <6E41111281623B4B8A9AB8F9A7EA34378244C9@wlmmsx02.nemours.org> > Content-Type: text/plain; charset=iso-8859-1 > > Does anyone know what I can use to get pararosaniline out of fabric? I was > going to try acid alcohol, but haven't tried anything yet out of fear of > making it worse. > > If anyone has suggestions, I and my brand new tan corduroy jack would much > appreciate it! > > Kristen > > kbroomal@nemours.org > > > > > > ------------------------------ > > Message: 4 > Date: Mon, 16 Jan 2006 08:59:12 -0500 > From: "Johns, Laura [CNTUS]" > Subject: [Histonet] Cell Pellets in Agar > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > <70F83FE9F65318468A612768E7043F8903753FE6@cntusmaexs9.na.jnj.com> > Content-Type: text/plain; charset="iso-8859-1" > > Does anyone have a protocol for paraffin embedding cell pellets? We've had > problems with cell pellets staying together after fixation and during > embedding, and I know that occasionally people use agar or agarose to keep > the pellets compact. Is it better to use agar or agarose? > > Thanks, > > Laura M. Johns, Ph.D. > Centocor, Inc. > 145 King of Prussia Road (Mail Stop # R-4-2) > Radnor, PA 19087 > Phone: 610-240-8284 .... Fax: 610-240-8150.... Email: > ljohns53@cntus.jnj.com > >> Confidentiality Notice: This message is intended for the individual(s) 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, he/she is >> 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 by telephone and delete >> the e-mail from your system immediately. Thank you. >> >> > > > ------------------------------ > > Message: 5 > Date: Mon, 16 Jan 2006 16:17:24 +0200 > From: "Sasa Jovanovic" > Subject: [Histonet] autofluorescence > To: > Message-ID: <000701c61aa7$9c07b810$640aa8c0@SinisaHome> > Content-Type: text/plain; charset="us-ascii" > > Hi everyone > Please help! In my project I am using immunohostochemistry to determine > presence/absence of integrins in rat uterine tissue. I am trying to make it > work for several months now with both alpha V beta 3 (immunoflorescence) and > alpha 4 beta 1 integrin (using DAKO ARK kit) and use of monoclonal > antibodies. My biggest problem so far is staining in the negative control > (positive and negative control stain exactly the same) and auto florescence. > I am following the Santa Cruz Protocol for immuno-fluorescence staining of > frozen tissue. I snap freeze rat uterine tissue in liquid nitrogen, cut 6 um > thin sections and adhere them on silane pre-treated slides and dry > overnight...... then I fix them in cold acetone for 10 minutes and proceed > with staining as follows: > - wash slides in three changes of PBS (pH9) > -incubate slides for 5-10 minutes in 0.1 H2O2 in PBS to quench endogenous > peroxidase activity > - wash slides in PBS 2x5minutes > - incubate slides with 10% normal goat blocking serum in PBS for 20 min to > suppress non - specific binding of Ig G > - wash in PBS (3x5min) > - incubate with primary antibody for 60 minutes > - wash in 3 changes of PBS > - incubate for 45 minutes with FITC conjugated secondary diluted in PBS with > 1.5-3% normal blocking serum in dark chamber > - wash in 3 changes of PBS > - mount and coverslip directly from PBS with aqueous mounting medium > Both primary (sc-7312)and secondary (sc-2010) antibody were purchased from > SCBT > I hope this is sufficient info to help you find some answers to my problem > Thank you in advance > Kind regards > Sasha > > > ------------------------------ > > Message: 6 > Date: Mon, 16 Jan 2006 14:27:15 -0000 > From: "Marshall Terry Dr, Consultant Histopathologist" > > Subject: RE: [Histonet] Dr Marshall & the English Language > To: , > Message-ID: > > Content-Type: text/plain; charset="iso-8859-1" > > Hmm. > Can't have it too pink means one likes it pink. I did not realise in using > this idiom that it is not universally understood. > That's the problem with idioms. > One does not ordinarily want to "detect eosinophilia". For that, the merest > whiff of eosin is what is needed, and that is too my eye, hopeless for > diagnostic work. > Perhaps Mike means that there is not the nuance of tint if overstained. Of > course. By overstating the case I was emphasising my preference to having a > blue and red stain to a blue and itsi bitsi pink-in-places stain:_) > > Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path > Consultant Pathologist > Rotherham General Hospital > South Yorkshire > England > terry.marshall@rothgen.nhs.uk > > -----Original Message----- > From: mtitford@aol.com [mailto:mtitford@aol.com] > Sent: 13 January 2006 18:06 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Dr Marshall & the English Language > > > Dr Terry Marshall sayeth about H & E's that "he cannot have it too pink". Does > he mean that he does not want it too pink, or that he likes it pink?! In any > event, how can you readily detect eosinophilia when the eosin is too heavy? > > Mike Titford > Pathology > USA Mobile AL USA > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 7 > Date: Mon, 16 Jan 2006 8:49:49 -0600 > From: Tyler W. > Subject: [Histonet] DI Water > To: histonet@lists.utsouthwestern.edu > Message-ID: <20060116144949.AB5D09E83C@merle.it.northwestern.edu> > Content-Type: text/plain > > The DI water tap here in the lab isn't working. Does anyone know if its > alright to use regular tap > water in a flotation bath? > > > > > > ------------------------------ > > Message: 8 > Date: Mon, 16 Jan 2006 08:27:35 -0700 > From: "Hansen, Donna \(Ontario\)" > Subject: RE: [Histonet] DI Water > To: > Message-ID: > <91FAC8D14C98974F839CA842BBC67D42136356@nwont2dc01.ontario-or.catholichealth.n > et> > > Content-Type: text/plain; charset="iso-8859-1" > > > > ________________________________ > > From: Hansen, Donna (Ontario) > Sent: Mon 1/16/2006 8:21 AM > To: Tyler W. > Subject: RE: [Histonet] DI Water > > > Good morning. > We have always used tap water in the water bath. flotation bath, water bath, > same thing? > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tyler W. > Sent: Mon 1/16/2006 7:49 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] DI Water > > > > The DI water tap here in the lab isn't working. Does anyone know if its > alright to use regular tap > water in a flotation bath? > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > ------------------------------ > > Message: 9 > Date: Mon, 16 Jan 2006 10:31:14 -0500 > From: "Monfils, Paul" > Subject: RE: [Histonet] Cell Pellets in Agar > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > <09C945920A6B654199F7A58A1D7D1FDE01717643@lsexch.lsmaster.lifespan.org> > > Content-Type: text/plain; charset="iso-8859-1" > > You can use a number of soluble, coagulable proteins to bind cell > pellets together, including agar, agarose, gelatin, albumin, serum, etc. I > use Mayer's albumin, but the technique is pretty much the same regardless of > which substance is used. After fixing the cells in suspension, I spin them > down and remove the fixative. Then wash a couple of times in buffer by > resuspending and spinning down. After the last wash I resuspend in buffer > once more, add 2 drops of Mayer's albumin per ml of suspension, mix > thoroughly by vortexing a few seconds, allow to stand for a couple of > minutes, then spin down once more and remove the supernatant. I then begin > dehydration of the pellet in the centrifuge tube, usually beginning with 50% > ethanol, then 70%, 95%, and absolute ethanol. Pipette the initial alcohol > gently down the side of the tube to ensure that the pellet is not disturbed. > Once you are in the stronger alcohols the thin coating of albumin on the > cells has coagulated, binding them firmly together. At some point in the > dehydration series the pellet, due to contraction during dehydration, > usually detaches from the tube intact. If it doesn't actually detach, it can > be detached either by drawing up alcohol into a pipette and squirting it > onto the pellet, or if necessary, by the slightest touch of a slender probe. > > I then either wrap the pellet in lens paper and place it in a > cassette on the tissue processor, starting with the final absolute alcohol > station, or, I finish the processing by hand, in the centrifuge tube. If > you are going to do the latter, make sure your tube is resistant to the > clearing agent. Polypropylene tubes work best for me. Polystyrene tubes > are more transparent, and are fine if you are only going to use alcohol in > the tube, but many clearing agents dissolve polystyrene. > > > >> ---------- >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of >> Johns, Laura [CNTUS] >> Sent: Monday, January 16, 2006 5:59 AM >> To: 'histonet@lists.utsouthwestern.edu' >> Subject: [Histonet] Cell Pellets in Agar >> >> Does anyone have a protocol for paraffin embedding cell pellets? We've >> had >> problems with cell pellets staying together after fixation and during >> embedding, and I know that occasionally people use agar or agarose to keep >> the pellets compact. Is it better to use agar or agarose? >> >> Thanks, >> >> Laura M. Johns, Ph.D. >> Centocor, Inc. >> 145 King of Prussia Road (Mail Stop # R-4-2) >> Radnor, PA 19087 >> Phone: 610-240-8284 .... Fax: 610-240-8150.... Email: >> ljohns53@cntus.jnj.com >> >>> Confidentiality Notice: This message is intended for the individual(s) >> 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, he/she is >>> 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 by telephone and delete >>> the e-mail from your system immediately. Thank you. >>> >>> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > > > ------------------------------ > > Message: 10 > Date: Mon, 16 Jan 2006 10:57:35 -0500 > From: "Monfils, Paul" > Subject: RE: [Histonet] Desperately needing suggestions please! > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > <09C945920A6B654199F7A58A1D7D1FDE01717644@lsexch.lsmaster.lifespan.org> > > Content-Type: text/plain; charset="iso-8859-1" > > For basic dyes like pararosaniline or hematoxylin, acid alcohol would > probably be your best bet. I keep a little bottle of acid alcohol (fairly > strong - about 4% hydrochloric acid in 70% ethanol) in the cabinet above the > washer at home, to pretreat such stains. Acid dyes like eosin or fast green > will usually wash out pretty easily without pretreatment because of the > basic pH of the tap water and the detergent. But those same basic > properties will set basic dyes rather than remove them. I just saturate the > stain with a little acid alcohol, work it in a bit, then blot it firmly > between paper towels (under and on top of the fabric) (pound on it a bit > actually), repeating if necessary, and then when I have sufficiently removed > the stain, drop the garment into the already running washer. Sounds like > your concern may be a non-washable garment, in which case the saturate and > blot technique alone will HOPEFULLY be sufficient. In that case I think I > would finish up with one last saturate and blot using either plain 70% > alcohol or distilled water, to remove excess acid from the fabric. > > I haven't seen a case where acid alcohol had any direct effect on fabric > color, but you might want to try it on an inconspicuous part of the garment > first, just to be sure. My wife once got some black grease on a white > blouse that had little green leaves and pink and blue flowers. I brought the > blouse in to work and treated it with xylene, which didn't completely remove > the grease. They I tried chloroform, which quickly removed the grease, and > the blue flowers. > > > > > ------------------------------ > > Message: 11 > Date: Mon, 16 Jan 2006 10:59:07 -0500 > From: "Helen Fedor" > Subject: RE: [Histonet] Cell Pellets in Agar > To: , > Message-ID: > Content-Type: text/plain; charset="us-ascii" > > Paul, Here is the protocol that we use in our Lab. > 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. > > > >>>> "Monfils, Paul" 01/16/06 10:31 AM >>> > You can use a number of soluble, coagulable proteins to bind cell > pellets together, including agar, agarose, gelatin, albumin, serum, etc. I > use Mayer's albumin, but the technique is pretty much the same regardless of > which substance is used. After fixing the cells in suspension, I spin them > down and remove the fixative. Then wash a couple of times in buffer by > resuspending and spinning down. After the last wash I resuspend in buffer > once more, add 2 drops of Mayer's albumin per ml of suspension, mix > thoroughly by vortexing a few seconds, allow to stand for a couple of > minutes, then spin down once more and remove the supernatant. I then begin > dehydration of the pellet in the centrifuge tube, usually beginning with 50% > ethanol, then 70%, 95%, and absolute ethanol. Pipette the initial alcohol > gently down the side of the tube to ensure that the pellet is not disturbed. > Once you are in the stronger alcohols the thin coating of albumin on the > cells has coagulated, binding them firmly together. At some point in the > dehydration series the pellet, due to contraction during dehydration, > usually detaches from the tube intact. If it doesn't actually detach, it can > be detached either by drawing up alcohol into a pipette and squirting it > onto the pellet, or if necessary, by the slightest touch of a slender probe. > > I then either wrap the pellet in lens paper and place it in a > cassette on the tissue processor, starting with the final absolute alcohol > station, or, I finish the processing by hand, in the centrifuge tube. If > you are going to do the latter, make sure your tube is resistant to the > clearing agent. Polypropylene tubes work best for me. Polystyrene tubes > are more transparent, and are fine if you are only going to use alcohol in > the tube, but many clearing agents dissolve polystyrene. > > > >> ---------- >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of >> Johns, Laura [CNTUS] >> Sent: Monday, January 16, 2006 5:59 AM >> To: 'histonet@lists.utsouthwestern.edu' >> Subject: [Histonet] Cell Pellets in Agar >> >> Does anyone have a protocol for paraffin embedding cell pellets? We've >> had >> problems with cell pellets staying together after fixation and during >> embedding, and I know that occasionally people use agar or agarose to keep >> the pellets compact. Is it better to use agar or agarose? >> >> Thanks, >> >> Laura M. Johns, Ph.D. >> Centocor, Inc. >> 145 King of Prussia Road (Mail Stop # R-4-2) >> Radnor, PA 19087 >> Phone: 610-240-8284 .... Fax: 610-240-8150.... Email: >> ljohns53@cntus.jnj.com >> >>> Confidentiality Notice: This message is intended for the individual(s) >> 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, he/she is >>> 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 by telephone and delete >>> the e-mail from your system immediately. Thank you. >>> >>> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 26, Issue 17 > **************************************** Aprill Watanabe, B.S. Research Associate Tissue Microarray Center (TMA) Translational Genomics Research Institute (TGen) 602-343-8822 awatanabe@tgen.org www.tgen.org From ree3 <@t> leicester.ac.uk Tue Jan 17 10:33:18 2006 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Tue Jan 17 10:33:32 2006 Subject: [Histonet] GCK-like kinase(GLK) Message-ID: --Anyone have any idea of the distribution of GLK in normal human tissues, unable to find anything in Pubmed. Thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER..U.K..... From SohrabB1 <@t> wmmcpo.ah.org Tue Jan 17 11:28:49 2006 From: SohrabB1 <@t> wmmcpo.ah.org (Behnaz Sohrab) Date: Tue Jan 17 11:29:48 2006 Subject: [Histonet] Histogel Message-ID: I would like to know if anyone out there using Histogel for the Nongyn using Thinprep? If yes, where do you get it ? Thanks, Behnaz Sohrab, Histo/Cyto Supervisor From plaurie <@t> benaroyaresearch.org Tue Jan 17 11:36:30 2006 From: plaurie <@t> benaroyaresearch.org (Patrick Laurie) Date: Tue Jan 17 11:36:50 2006 Subject: [Histonet] Cryosectioning mouse Eyeball Message-ID: <2ff462f0148b4f45b9a340e105039838@penguin.vmresearch.org> I have a couple of Researchers here who are interested in cryosectioning mouse eyeballs to look at the corneas. I have given them a few hints (colder temperatures, cutting quickly and evenly, etc.) but they are having variable results. I was wondering if anyone has any further hints. Is there some sort of freezing media that someone would recommend? We are using Tissue-Tek OCT. Thanks, Patrick Laurie, HT Benaroya Research Institute 1201 9th Ave Seattle, Wa 98101 (206) 341-0681 From jkiernan <@t> uwo.ca Tue Jan 17 11:41:28 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Tue Jan 17 11:41:30 2006 Subject: [Histonet] Dry Stains Shelf Life References: <43CCE44E.7090107@bms.com> Message-ID: <43CD2C48.F22D41E4@uwo.ca> See the following publications: Emmel VM & Stotz EH 1986. Certified biological stains: a stability study. Stain Technol. 61:385-387. Titford M 2001. Comparison of historic Grubler dyes with modern counterparts. Biotech. Histochem. 76:23-30. Titford, M. 2002. Save that dye! Microscopy Today 18-5:31-34. The general conclusion is that nearly all dye powders still work after 100 years (Titford). Quantitative assays show very little deterioration over 20 or so years (Emmel & Stotz). Matt Frank may have more recent information from the Biological Stain Commission's lab. Alcian blue 8G can deteriorate in the solid state, changing to an insoluble pigment. This is not a problem with the pyridine variant of alcian blue (Churukian et al 2000, Biotech. Histochem. 75: 147-150). Acidic solutions of alcian blue 8G are stable for some years on my shelves. Churukian's lab manual gives a recommended shelf life of 6 months. An alcian blue solution with a precipitate should be discarded and replaced, not filtered and used. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Sharon E Willman wrote: > > Hi, > What is the expiration time for dry stains and how often should you > replenish? Many times they do not have expiration dates on them when > they come in from the manufacturer. > I would appreciate any information on this matter. > Thanks, > Sharon Willman > From jkiernan <@t> uwo.ca Tue Jan 17 11:55:20 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Tue Jan 17 11:55:20 2006 Subject: [Histonet] paraformaldehyde crystals v Prills References: <001701c61b44$6d730b60$112b5c9f@Carlos> Message-ID: <43CD2F88.7CA05654@uwo.ca> Carl Hobbs wrote: > > Many of our Groups use paraformaldehyde crystals ( Sigma P6148) to make up > their solution of formalin. Works very well; 4g + 1tab PBS + 6microlitre > 10MNaOH.....stir overnight. Dissolved completely:pH7.2 > I am trying to get away from using this paraformaldehyde: it is very light > and prone to spread around the fume hood when it's weighed out. > So, I bought PRILLS from Sigma:Problem is, they just do not dissolve > completely, unless I preboil the water.( heating to 60C is not hot enough to > dissolve the Prills. > I have tried all combinations of the formulation above( eg: no NaOH, > dissolving PBS tablet before adding paraformaldehyde). > I want the procedure with the least risk; boiling may be a higher risk than > the spread of powder. > NB: I cannot turn down/off the fume hoods, for the weighing of the > crystals( which behave more like a very fine powder), hence my desire to use > the heavier Prills. > > Be grateful for any insights. > Carl You can avoid the fine powder and insolubility problems by making your fixative from formalin rather than paraformaldehyde. A good mixture, shown to be OK even for electron microscopy, is that of Carson FL, Martin JH & Lynn JA 1973, Am. J. Clin. Path. 59:365-373. Its composition is: Formalin 100 ml Water 900 ml Monobasic sodium phosphate, monohydrate 18.6 g Sodium hydroxide 4.2 g The pH is 7.2-7.4 and the solution can be kept for several months. As always, don't rely on the accuracy of this or any other email! Consult the original paper before making and using the fixative. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ From kbroomal <@t> NEMOURS.ORG Tue Jan 17 12:44:29 2006 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Tue Jan 17 12:44:49 2006 Subject: [Histonet] Desperately needing suggestions please! Message-ID: <6E41111281623B4B8A9AB8F9A7EA34378244CC@wlmmsx02.nemours.org> A big thank you to everyone who sent stain buster techniques to me. I'll let you know what works (or doesn't work if the case may be!). Thanks again!! Kristen kbroomal@nemours.org From sjchtascp <@t> yahoo.com Tue Jan 17 13:28:41 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Tue Jan 17 13:28:52 2006 Subject: [Histonet] decal- processing Message-ID: <20060117192841.40119.qmail@web90208.mail.scd.yahoo.com> I just finished decaling some bone joints in formic acid. Should my processing cycle begin with a low graded alcohol like 70% or would it be ok to run them through the formalins. I know with HCL based decalcifiers the fprmer would be the way to go after rinsing in water. This is the 1st time I've used formic acid. Steve --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. From lhotaks <@t> mcmaster.ca Tue Jan 17 14:18:38 2006 From: lhotaks <@t> mcmaster.ca (Sarka Lhotak) Date: Tue Jan 17 14:18:48 2006 Subject: [Histonet] Loosing NoveRed signal Message-ID: Hello Histonetters, this is a very annoying problem that I see intermitently. While developing a colour with NovaRed, I can see a nice staining. I stop the reaction with distilled water, do several changes in distilled and let sit for 10 minutes. Then Hematoxylin, alcohols, xylenes and coverslipping in Permount. Once counterstained and coverslipped, the staining is gone! THis problem is not very frequent but all the more frustrating. Once I talked to Vector about this and was advised to let the slides sit in water for 10 minutes before hematoxylin. But it does not always help. Since the problem is intermittent I still have not figured out what is happening. It is very frustrating, you see a great staining but then it is gone and nobody will believe you! Has anybody experienced this and have a remedy? Thanks for your help, Sarka Lhotak Henderson Research Centre, McMaster University, Hamilton, Ontario From bwhitaker <@t> brownpathology.com Tue Jan 17 14:36:51 2006 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Tue Jan 17 14:36:59 2006 Subject: [Histonet] Loosing NoveRed signal In-Reply-To: Message-ID: <000101c61ba5$bf437220$3601a8c0@brownpathology.net> Hi Sarka, I do know that some chromogens, while advertised as permanent, can fade completely in exposure to alcohol. (When I was a Biotek Solutions rep, a customer left his slides in alcohol while he went to lunch, and when he returned, the color had completely faded.) I don't know about this particular chromogen, but it is something to keep in mind. Do you always quickly dehydrate and clear, or are there some days when the slides are in alcohol longer than other days? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sarka Lhotak Sent: Tuesday, January 17, 2006 2:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Loosing NoveRed signal Hello Histonetters, this is a very annoying problem that I see intermitently. While developing a colour with NovaRed, I can see a nice staining. I stop the reaction with distilled water, do several changes in distilled and let sit for 10 minutes. Then Hematoxylin, alcohols, xylenes and coverslipping in Permount. Once counterstained and coverslipped, the staining is gone! THis problem is not very frequent but all the more frustrating. Once I talked to Vector about this and was advised to let the slides sit in water for 10 minutes before hematoxylin. But it does not always help. Since the problem is intermittent I still have not figured out what is happening. It is very frustrating, you see a great staining but then it is gone and nobody will believe you! Has anybody experienced this and have a remedy? Thanks for your help, Sarka Lhotak Henderson Research Centre, McMaster University, Hamilton, Ontario _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From info <@t> instrumedics.com Tue Jan 17 14:45:29 2006 From: info <@t> instrumedics.com (Instrumedics) Date: Tue Jan 17 14:45:42 2006 Subject: [Histonet] Cryosectioning mouse Eyeball References: <2ff462f0148b4f45b9a340e105039838@penguin.vmresearch.org> Message-ID: <011501c61ba6$fd163550$6401a8c0@INSTRUMEDICS22> Patrick, Preparing frozen sections of mouse eyeballs is difficult, especially if you would like to retain the morphology of the structures of interest. We have been successful using the CryoJane Tape-Transfer system. Please visit the Instrumedics' web site for the details on the CryoJane method. If you have any questions please contact us. Bernice Instrumedics ----- Original Message ----- From: "Patrick Laurie" To: Sent: Tuesday, January 17, 2006 12:36 PM Subject: [Histonet] Cryosectioning mouse Eyeball I have a couple of Researchers here who are interested in cryosectioning mouse eyeballs to look at the corneas. I have given them a few hints (colder temperatures, cutting quickly and evenly, etc.) but they are having variable results. I was wondering if anyone has any further hints. Is there some sort of freezing media that someone would recommend? We are using Tissue-Tek OCT. Thanks, Patrick Laurie, HT Benaroya Research Institute 1201 9th Ave Seattle, Wa 98101 (206) 341-0681 From Kristopher.Kalleberg <@t> unilever.com Tue Jan 17 14:52:59 2006 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Tue Jan 17 14:53:09 2006 Subject: [Histonet] malondialdehyde protocol Message-ID: All, Does anyone have a malondialdehyde protocol that seems to work pretty well. Thanks. Kris From Eric <@t> ategra.com Mon Jan 16 19:49:19 2006 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Tue Jan 17 14:58:56 2006 Subject: [Histonet] Histotech opportunities-Bench and Supervisory-All States Available-Permanant and Temporary Message-ID: Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. Most of my Histo Techs positions are clinical, although I do have a few Temporary and Research Histo Tech positions as well. 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Florida (South) (Full-time, Perm, Bench) The clients are currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recruiter 1-800-466-9919 ext 223 --------------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu ---------------------------------------------------------------- ---------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. ---------------------------------------------------------------- From portera <@t> msu.edu Tue Jan 17 15:19:01 2006 From: portera <@t> msu.edu (Amy Porter) Date: Tue Jan 17 15:20:07 2006 Subject: [Histonet] Loosing NoveRed signal References: Message-ID: <002701c61bab$a3157d90$8e7a0923@HistoJJ> Sarka - We use Nova Red in our laboratory on a regular basis and do not encounter this problem. However, I do know if you review the product insert it will tell you that if you use Deionized water opposed to glass distilled it will inhibit the reaction. "Deionized water may contain inhibitors of the peroxidase reaction". We rinse our slides in distilled water and counterstain with hematoxylin, dehydrate, clear in xylene. We have even left them overnight in xylene to see if the reaction is removed and it has not been a problem. I would think it might be connected to the water somehow. ----- Original Message ----- From: "Sarka Lhotak" To: Sent: Tuesday, January 17, 2006 3:18 PM Subject: [Histonet] Loosing NoveRed signal > Hello Histonetters, > this is a very annoying problem that I see intermitently. While > developing a colour with NovaRed, I can see a nice staining. I stop the > reaction with distilled water, do several changes in distilled and let > sit for 10 minutes. Then Hematoxylin, alcohols, xylenes and > coverslipping in Permount. Once counterstained and coverslipped, the > staining is gone! THis problem is not very frequent but all the more > frustrating. > Once I talked to Vector about this and was advised to let the slides > sit in water for 10 minutes before hematoxylin. But it does not always > help. Since the problem is intermittent I still have not figured out > what is happening. It is very frustrating, you see a great staining but > then it is gone and nobody will believe you! Has anybody experienced > this and have a remedy? > Thanks for your help, > > Sarka Lhotak > > Henderson Research Centre, McMaster University, > Hamilton, Ontario > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Kim.Osullivan <@t> med.monash.edu.au Tue Jan 17 15:44:15 2006 From: Kim.Osullivan <@t> med.monash.edu.au (Kim O'Sullivan) Date: Tue Jan 17 15:48:29 2006 Subject: [Histonet] fixation Message-ID: <130.194.114.210.1137533976@my.monash.edu.au> Hi all, Have recieved some mouse kidneys recently to work on which have been fixed in PLP for aproximatley 2 hours and 7% sucrose for 2 days afterwards (5 changes), but the glomeruli appear shrunken and the bowmans space enlarged (not a product of the disease we are researching). Is this caused by overfixing or underfixing? Alternatively (as this method is quite common and we have not had this problem in the past) could this result occur throught the dehyration or rehydration steps during the staining process? Any ideas would be helpful Kim O'Sullivan From dw18 <@t> uchicago.edu Tue Jan 17 16:15:08 2006 From: dw18 <@t> uchicago.edu (David A. Wright) Date: Tue Jan 17 16:15:29 2006 Subject: [Histonet] RE: paraformaldehyde crystals v Prills Message-ID: <3f8d1bb2.7c3e3f69.81b7c00@m4500-03.uchicago.edu> In reply to: Histonet Digest, Vol 26, Issue 19 Message: 2 Hello All It may not help Carl in the UK (I see you are at King's College, London) but we have tried various suppliers' paraformaldehyde and much prefer Fisher Scientific's own brand (T353-500) precisely because it is a fine prill - particulate enough not to blow away easily but fine enough to dissolve "easily". In my experience though, no suppliers' paraformaldehyde ever disssolves completely (there is always a fine haze,presumably of high MW polymer) in a timescale that I find acceptable. It will settle out, so if you are happy to decant/pipet off the clear supernatant the next day, that may be ok, but I use it for brain perfusions and worry about clogged capillaries. I always vacuum filter it through a Buechner Funnel but this is time consuming and takes a lot of filters unless you want to watch your formaldehyde de-gas down the vacuum line. So for this reason alone (avoiding filtration) it is probably a very good idea to follow John Kiernan's formalin suggestion, especially if you only need a small amount. This, however raises the question of why you are repeatedly making small amounts if you don't like messing with the powder. Contrary to rumo[u]r, buffered paraformaldehyde keeps very well (as Dr Kiernan has pointed out - see the archives); I have often used it after several months in the cold and found the fixation to be excellent. Just check the pH first. A second point is that, from the variations in the protocol he tried, I'm not sure that Carl understands the desirability of using NaOH in the prep. The paraformaldehyde polymer hydrolyses faster at basic than neutral pH. If a small amount of alkali is added simultaneously with or after the PBS tablet, the pH will not be basic enough to have this effect. The NaOH must be added first and the PBS only added once the PFA is in solution. I prefer to use dibasic sodium or potassium phosphate as this directly gives a basic solution (pH 10-11) in which the PFA dissolves easily (15-20 min at ca. 50?C) and then I neutralize with phosphoric acid once the solution clears. I hope this helps cheers -David David A. Wright. Ph.D The University of Chicago/Neurosurgery ======================================================= Date: Tue, 17 Jan 2006 12:55:20 -0500 From: "John A. Kiernan" Subject: Re: [Histonet] paraformaldehyde crystals v Prills To: Carl Hobbs Cc: histonet@lists.utsouthwestern.edu Message-ID: <43CD2F88.7CA05654@uwo.ca> Content-Type: text/plain; charset=us-ascii Carl Hobbs wrote: Many of our Groups use paraformaldehyde crystals ( Sigma P6148) to make up their solution of formalin. Works very well; 4g + 1tab PBS + 6microlitre 10MNaOH.....stir overnight. Dissolved completely:pH7.2 I am trying to get away from using this paraformaldehyde: it is very light and prone to spread around the fume hood when it's weighed out. So, I bought PRILLS from Sigma:Problem is, they just do not dissolve completely, unless I preboil the water.( heating to 60C is not hot enough to dissolve the Prills. I have tried all combinations of the formulation above( eg: no NaOH, dissolving PBS tablet before adding paraformaldehyde). I want the procedure with the least risk; boiling may be a higher risk than the spread of powder. NB: I cannot turn down/off the fume hoods, for the weighing of the crystals( which behave more like a very fine powder), hence my desire to use the heavier Prills. Be grateful for any insights. Carl You can avoid the fine powder and insolubility problems by making your fixative from formalin rather than paraformaldehyde. A good mixture, shown to be OK even for electron microscopy, is that of Carson FL, Martin JH & Lynn JA 1973, Am. J. Clin. Path. 59:365-373. Its composition is: Formalin 100 ml Water 900 ml Monobasic sodium phosphate, monohydrate 18.6 g Sodium hydroxide 4.2 g The pH is 7.2-7.4 and the solution can be kept for several months. As always, don't rely on the accuracy of this or any other email! Consult the original paper before making and using the fixative. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ From jstaruk <@t> masshistology.com Tue Jan 17 16:19:50 2006 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Tue Jan 17 16:19:54 2006 Subject: [Histonet] Pre-collagen question In-Reply-To: <130.194.114.210.1137533976@my.monash.edu.au> Message-ID: <000001c61bb4$22c6b970$6500a8c0@FrontOffice> Hi all, I received a request to perform a "pre-collagen" stain on skin tissue using a version of Herovici's stain for young and mature collagen (mature collagen stains red while young collagen stains blue). For a control, I used a 5-day old post surgical incision which demonstrates nice granulation tissue and fibroblasts forming connective tissue at the incision and around the suture material. There's plenty of red collagen but nothing is staining blue. Can anyone advise me if this is the appropriate control to use for this pre-collagen stain? Anyone planning on invoicing me for their reply need not respond to this question. Thank you Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com From JMacDonald <@t> mtsac.edu Tue Jan 17 23:28:55 2006 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Tue Jan 17 23:28:00 2006 Subject: [Histonet] RE: Eosin too pink Message-ID: Eosin is differentiated in the dehydrating alcohols. You ca the time in the alcohols to get a lighter pink. Also contact tech support at Richard-Allan. There are two types of eosin that they sell and one is really concentrated. Jennifer -----histonet-bounces@lists.utsouthwestern.edu wrote: --- To: From: Mark Adam Tarango To: histonet@lists.utsou > Subject: [Histonet] Eosin too pink > > H > We are currently doing our H & E stain by hand an > Richard-Allan 7211 Hematoxylin and their Eosin Y. pathologist is > still complaining of slides being TOO PINK. &nb the eosin step > down to just 5 dips from 15 dips, and Eosin is very > rich. > > Thi > Hydrate to water like normal (thru Clear Rite 3, Flex) > Hematoxylin- 2 mins > Rinsing in water u > Clarifier II- 30 dips > running wat > Richard Allan Bluing - 30 dips > running water - > 95% Flex - 10 dips > Eosin-Y- 5 quick dips > > Any suggestions on a pos Eosin. > Complaint seems to solely experimenting w/ > cutting the eo but, the > Hemo looks if that > will test slides. > > > Helayne Parker > Histology Sectio > Skaggs Community Health Center > Branson, Missouri ______________________ 5F ______________________ 5F_ Histonet mailing list Histonet@lists.utsouthwestern.edu [1] References 1. file://localhost/tmp/3D"h From jkiernan <@t> uwo.ca Wed Jan 18 00:17:06 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Wed Jan 18 00:16:35 2006 Subject: [Histonet] fixation [of mouse kidney; long reply] References: <130.194.114.210.1137533976@my.monash.edu.au> Message-ID: <43CDDD62.ABF5E540@uwo.ca> You cannot overfix tissue structure with formaldehyde. You can interfere with chemical properties of the tissue. For example, storage in formalin for many years causes loss of stainability with eosin. Assuming that your PLP is the periodate-lysine-"paraformaldehyde" mixture of McLean & Nakane (1974): this was devised for preserving cell-surface carbohydrates (glycocalyx) for EM immunohistochemistry. It doesn't work the way the originators intended because the lysine and formaldehyde react rapidly in the solution, forming polymers. The polymers may be able to form long cross-links, but they cannot penetrate easily through cell membranes. Interestingly, periodate alone can cross-link cell surface glycoproteins. See Hixson et al 1981 J. Histochem. Cytochem. 29: 561-566 for an important study of periodate-lysine-"paraformaldehyde". The paper is available free of charge at http://www.jhc.org/content/vol29/issue4/ Anyone can download the whole paper (.pdf) or just the abstract of any item in the JHC archives that's more than about a year old. Anyone intending to use PLP (periodate-lysine-"paraformaldehyde") should read this paper. Returning to your mouse kidneys! They may be inadequately fixed, with the glomeruli shrinking within Bowman's capsule, perhaps an effect of the hypertonic sucrose cryoprotectant. Intuitively, that's the simplest explanation if your PLP is indeed periodate-lysine-"paraformaldehyde". BUT if you're accustomed to looking at sections of well fixed rats' kidneys, there is a difference in the anatomy of the glomeruli. In the mouse, the glomerular tuft of capillaries is squat, with a wide base that's usually in the plane of section. Rat glomeruli are more like human ones, with the tuft circular in section and its narrow base (origin) visible in only a minority of sectioned glomeruli. In a well fixed kidney, the great majority of glomeruli have very little empty space between the tuft of capillaries and Bowman's capsule. That's supposed to be closest approximation to the condition in life (see histology and histotechnology textbooks etc). There are a few lousy fixatives that inflate the glomerular tuft and break Bowman's capsule - an obvious artifact. See Histochemical Journal 17:1131-1146 (1985) for some pictures of badly fixed kidneys. John Kiernan Anatomy, UWO London, Canada. -------------------------------------------- Kim O'Sullivan wrote: > > Hi all, > > Have recieved some mouse kidneys recently to work on which have been fixed in PLP for aproximatley 2 hours and 7% sucrose for 2 days afterwards (5 changes), but the glomeruli appear shrunken and the bowmans space enlarged (not a product of the disease we are researching). Is this caused by overfixing or underfixing? > > Alternatively (as this method is quite common and we have not had this problem in the past) could this result occur throught the dehyration or rehydration steps during the staining process? > > Any ideas would be helpful > > Kim O'Sullivan > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From L.Driessen <@t> orthop.umcn.nl Wed Jan 18 00:21:37 2006 From: L.Driessen <@t> orthop.umcn.nl (L.Driessen@orthop.umcn.nl) Date: Wed Jan 18 00:21:47 2006 Subject: [Histonet] Re: Protocol for GMA Message-ID: <2E2AC813A84E054C8E8E572131082BE2145521@umcnet14.umcn.nl> Dear Mei Yin, You can find protocols on the internet. Just search for 'technovit 7100 GMA protocol' or something like that and you'll get some answers. Since you already fix the speciments in aceton then you can go to straigth on to GMA if you like. I would suggest that you first use a 1:1 mixture of aceton and GMA before going to pure GMA L?on Driessen UMCN St. Radboud Nijmegen The Netherlands From carl.hobbs <@t> kcl.ac.uk Wed Jan 18 08:17:08 2006 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Wed Jan 18 08:17:47 2006 Subject: [Histonet] re: paraformaldehyde crystals v Prills Message-ID: <002901c61c39$ddff2f80$112b5c9f@Carlos> Thank you for the three replies. Much appreciated. My apologies for any confusion caused: I regularly use a Formalin fixing solution derived from the concentrated Formalin solution. I also need to use a Formalin fixing solution derived from Paraformaldehyde for certain immunostaining instances. I do make up the latter in bulk and freeze in the small aliquots required for T/C cell monolayer fixing. I don't have any problem making up either of the solutions. The latter dissolves completely. What I really want to do is to use Prills instead of paraformaldehyde crystals. Unfortunately, they do not completely dissolve - around 80% disssolved after 24hrs. Sure , I can filter afterwards, but, I want to establish the SOP with the least risk/handling . Hence my request for help. Does anyone have any experience of using Prills? Thanks for the "Fischer" suggestion....I will check them out over here. Best wishes carl From Tiffany.L.Sheffield <@t> uth.tmc.edu Wed Jan 18 09:40:57 2006 From: Tiffany.L.Sheffield <@t> uth.tmc.edu (Sheffield, Tiffany L) Date: Wed Jan 18 09:41:01 2006 Subject: [Histonet] IHC & Plastic Section Question Message-ID: Hi Histonetters! Now please nobody laugh at me, but has anyone done any IHC in GMA (Technovit 7200) embedded sections of bone? I know there are some IHC techniques in MMA. I have been playing around with some techniques but I am unable to find any literature. To my knowledge in this particular plastic it has not been done, I could be wrong of course, that is why I am asking you all. Any help would greatly be appreciated. Tiffany Sheffield-Lopez, HT(ASCP) Supervisor, Bone Histomorphometry & Biomaterials Lab Department of Orthopaedic Surgery UTHSC-Houston Medical School 6431 Fannin, Suite 6.144 MSB Houston , TX 77030 713-500-6803 WK 713-500-0729 Fax _______________________________________________ From mhorne <@t> upei.ca Wed Jan 18 10:08:11 2006 From: mhorne <@t> upei.ca (Margaret Horne) Date: Wed Jan 18 11:05:28 2006 Subject: [Histonet] cryo on crustaceans Message-ID: <43CE2FAB.12875.C5076C@localhost> Hello All , We have two grads in our dept about to do cryo on Snow crab and Sea Lice. They are trying dif protocols but I said I said I would ask if any of you have done cryo on crustaceans. Thanks , Margaret Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada From pathrm35 <@t> adelphia.net Wed Jan 18 11:10:53 2006 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Wed Jan 18 11:11:03 2006 Subject: [Histonet] seeking employment Message-ID: <12649220.1137604253443.JavaMail.root@web30> Fellow Techs, I am seeking an employment opportunity as an assistant supervisor or lead histotech. I prefer private labs and specialize in dermpath and IHC. I am willing to work any shift and will relocate for the right opportunity. Thanks in advance, Ron Martin, BS, HT (ASCP) HTL, QIHC Florida licensed Histology Supervisor From HornHV <@t> archildrens.org Wed Jan 18 11:45:26 2006 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Wed Jan 18 11:44:58 2006 Subject: [Histonet] coverslipper Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDF55@EMAIL.archildrens.org> When I was NSH this past year, there was a coverslipper there that had the mounting media directly on the coverslips and a solution activated it to mount it to the slide. I cannot remember the company who made this. If you know would you please email me or if you are the company who makes this coverslipper contact me please. Thanks. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From MVaughan4 <@t> ucok.edu Wed Jan 18 13:47:17 2006 From: MVaughan4 <@t> ucok.edu (MVaughan4@ucok.edu) Date: Wed Jan 18 13:47:55 2006 Subject: [Histonet] IHC losing tissue In-Reply-To: Message-ID: Sharon, About your IHC slides losing tissues during processing: It's probably what Rene and Linda said - preheating is necessary. But I have heard that the treated slides purchased from vendors lose their stickiness over time. You might check other lots or other vendors to see if you still have problems. Were these slides brand new? Has anybody else had problems with old pretreated slides losing their stickiness? Mel Melville B. Vaughan, Ph. D. Assistant Professor Department of Biology University of Central Oklahoma Edmond, OK 73034 From PIXLEYSK <@t> UCMAIL.UC.EDU Wed Jan 18 13:59:46 2006 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Wed Jan 18 13:59:50 2006 Subject: [Histonet] RE: Histonet Digest, Vol 26, Issue 20 Message-ID: Dear John Kiernan and histonetters: I read with great interest the article you suggested on the PLP fixative. And I searched PubMed for similar articles. But I do not see a good analysis of what might be the alternative to PLP. The authors of the paper, Hixson et al., suggest that paraformaldehyde and lysine might be just as effective as PLP, with lowered cell toxicity, but they don't do an analysis of the effects on morphology or antigenicity. Have you tried anything like this? Does a combo of PF and lysine provide a better fixation of carbohydrate antigens than PLP? Do you know if these authors or anyone else followed up on this and developed and described a better fixative in more detail? The reason that I ask is that I have an antigen that is particularly sensitive to fixation and the monoclonal antibody to it works only with the PLP fixative. Supposedly the antigen is the carbohydrate portion of a cell surface receptor. But the fixation is so light that I lose almost all my cultured cells during processing for IHC (I have very lightly attached cultured cells). I have been searching for an alternative to PLP for years! So I thank you greatly for this article. I guess my next step would be to try different concentrations of paraformaldehyde and lysine. But if this has been tried before, I would appreciate any suggestions! Sarah Pixley From jkiernan <@t> uwo.ca Wed Jan 18 14:22:02 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Wed Jan 18 14:22:04 2006 Subject: [Histonet] Re: Histonet Digest, Vol 26, Issue 20 References: Message-ID: <43CEA36A.A6B0A85@uwo.ca> If your antigen is damaged by fixatives that contain formaldehyde, and you're not doing electron microscopy, why not try a simple coagulant fixative such as Clarke's or Carnoy's fluid or Puchtler's methacarn? These are good for conventional carbohydrate histochemistry (except glycolipids, of course). Check first that your cell culture plasticware is not soluble in the fixative. For a thin layer of cells methanol alone may be OK - as when fixing a blood film. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Pixley, Sarah (pixleysk)" wrote: > > Dear John Kiernan and histonetters: > > I read with great interest the article you suggested on the PLP > fixative. And I searched PubMed for similar articles. But I do not see a > good analysis of what might be the alternative to PLP. The authors of > the paper, Hixson et al., suggest that paraformaldehyde and lysine might > be just as effective as PLP, with lowered cell toxicity, but they don't > do an analysis of the effects on morphology or antigenicity. Have you > tried anything like this? Does a combo of PF and lysine provide a better > fixation of carbohydrate antigens than PLP? Do you know if these > authors or anyone else followed up on this and developed and described a > better fixative in more detail? > > The reason that I ask is that I have an antigen that is particularly > sensitive to fixation and the monoclonal antibody to it works only with > the PLP fixative. Supposedly the antigen is the carbohydrate portion of > a cell surface receptor. But the fixation is so light that I lose almost > all my cultured cells during processing for IHC (I have very lightly > attached cultured cells). I have been searching for an alternative to > PLP for years! So I thank you greatly for this article. I guess my next > step would be to try different concentrations of paraformaldehyde and > lysine. But if this has been tried before, I would appreciate any > suggestions! > > Sarah Pixley From JGordon <@t> cellmarque.com Wed Jan 18 14:24:12 2006 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Wed Jan 18 14:24:22 2006 Subject: [Histonet] IHC losing tissue References: Message-ID: Cell Marque promotes using pressure cookers with our Trilogy EDTA solution, which is a pretty aggressive pretreatment. We use this with pretty much all of our tissues, including breast and prostate needle biopsies and skin and other sensitive tissues. To do this, it is essential to prepare the cut tissue properly on the slides to ensure that no tissue is lost. In some cases, if you are using higher pH solutions for heat retrieval, the tissue preparation still can't prevent the tissue from coming off, which is why oftentimes people will move from high pH solutions to Trilogy, which doesn't have the higher pH but still gives comparable staining. This is our standard method of slide preparation: 1) Cut tissue 3-4 microns. This is sufficient for clinical IHC. Anything thicker runs a much higher risk of tissue loss. 2) Use positively charged slides that have been handled carefully (only touch the label area or the edges, do not touch the face of the slide as it may draw the charge off the slide). 3) Dry the slides for at least 2 hours at 58 degrees Celsius in a slide drying oven. For fatty breast, extend the drying time to 4 hours. This ensures that all water will escape from underneath the melted paraffin and tissue, and allow the tissue to form an uninhibited bond with the slide. If you have to dry the tissue faster, you can also use a microwave method. Methods used with the microwave for slide drying include running the microwave for 1.5 minutes, then allowing it to sit for one minute for the paraffin to congeal, and then run it for another 1.5 minutes, and then let sit again for a minute. The slide is ready for heat retrieval now and should give you good tissue adhesion. By using that method, we don't have to use any additives in our waterbath for tissue adhesion. Jeff Gordon Cell Marque Corp. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of MVaughan4@ucok.edu Sent: Wed 1/18/2006 1:47 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] IHC losing tissue Sharon, About your IHC slides losing tissues during processing: It's probably what Rene and Linda said - preheating is necessary. But I have heard that the treated slides purchased from vendors lose their stickiness over time. You might check other lots or other vendors to see if you still have problems. Were these slides brand new? Has anybody else had problems with old pretreated slides losing their stickiness? Mel Melville B. Vaughan, Ph. D. Assistant Professor Department of Biology University of Central Oklahoma Edmond, OK 73034 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RBARNHART <@t> summithealth.org Wed Jan 18 14:25:52 2006 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Wed Jan 18 14:26:11 2006 Subject: [Histonet] Histology standard Message-ID: A while back there was a study about the average time spent performing the different task in histology. I have tried to go onto The Journal Of Histotechnology and reprint it but it is not there. If there is anyone that has it that can send it to me it would be greatly appreciated. Thanks in advanced! Becky Barnhart, HT (ASCP) Histology Section Head From rjbuesa <@t> yahoo.com Wed Jan 18 14:40:53 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Jan 18 14:41:03 2006 Subject: [Histonet] IHC losing tissue In-Reply-To: Message-ID: <20060118204053.58482.qmail@web61219.mail.yahoo.com> Melville: You have a very good point there! I have been also told that the (+) slides after their expiration date can lose their properties. That fact did not come to my mind becasuse we were never confronted with it since we used all our (+) slides before the expiration date. This is something that should be considered also. Ren? J. MVaughan4@ucok.edu wrote: Sharon, About your IHC slides losing tissues during processing: It's probably what Rene and Linda said - preheating is necessary. But I have heard that the treated slides purchased from vendors lose their stickiness over time. You might check other lots or other vendors to see if you still have problems. Were these slides brand new? Has anybody else had problems with old pretreated slides losing their stickiness? Mel Melville B. Vaughan, Ph. D. Assistant Professor Department of Biology University of Central Oklahoma Edmond, OK 73034 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. From acjanes <@t> bu.edu Wed Jan 18 18:08:31 2006 From: acjanes <@t> bu.edu (acjanes@bu.edu) Date: Wed Jan 18 18:09:23 2006 Subject: [Histonet] ICC Background Message-ID: <20060118190831.8ggkko488cw84swo@www.bu.edu> Hi, I really need some suggestions about the background I am getting when running immunocytochemistry. I am using mouse brain tissue that is 30microns thick and has been perfused with 4% para. We have used different primaries and secondaries and we always seem to get really high background regardless of which primary or secondary we are using. I have also gotten high background in my non-primary control. The background is so high I can't even tell if there is any specific binding. I have run ICC many times in the past and I have gotten it to work before. I am not sure what is going on so if anyone has any suggestions please let me know! Thanks! Amy From linresearch <@t> aol.com Wed Jan 18 18:18:10 2006 From: linresearch <@t> aol.com (linresearch@aol.com) Date: Wed Jan 18 18:18:26 2006 Subject: [Histonet] Negative Serum Message-ID: <8C7EAA4F6E00C28-1234-37A@mblk-d51.sysops.aol.com> Hello, I need to prepare my own negative rabbit IgG for negative control FFPE tissue staining. I have tried using the rabbit negative Ab that I bought but it seems to be giving some positive staining. I would appreciate any advice on dilutions and diluent. Lin From sweaver <@t> bbpllab.com Wed Jan 18 18:22:49 2006 From: sweaver <@t> bbpllab.com (Steve Weaver) Date: Wed Jan 18 18:22:59 2006 Subject: [Histonet] DIF Controls Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A02E2EB@bbplsrv1.bbpl> **New** CAP Question: ANP.21850 "Are appropriate positive and negative controls performed with each case tested using immunofluorescence?" We currently just run one (+) control, usually IgG or C3, for the entire run that includes IgA, IgG, IgM, C3, C4 and fibrinogen. My question is - should I be running (+) controls for all antibodies? Your thoughts please. Please indicate if you have been inspected on this new question. Thank you. sw From emry <@t> u.washington.edu Wed Jan 18 18:37:53 2006 From: emry <@t> u.washington.edu (Trisha Emry) Date: Wed Jan 18 18:36:17 2006 Subject: [Histonet] tooth curl Message-ID: I just did serial sections through a decaled, paraffin embedded tooth. I had trouble keeping sections stained with H & E on "plus" slides and they often curled up at the edges and folded. Any advice on how to deal with the ones cut and/or future teeth? Thanks, Trisha U of Washington, Seattle From hll0211 <@t> aol.com Wed Jan 18 20:10:16 2006 From: hll0211 <@t> aol.com (hll0211@aol.com) Date: Wed Jan 18 20:10:41 2006 Subject: [Histonet] New to Histonet Message-ID: <8C7EAB49F95F6B1-F58-54A0@FWM-D31.sysops.aol.com> My name is Hannah Long and I from Texarkana, AR. I am currently enrolled in a Vertebrate Histology class at Texas A&M - Texarkana. I have subscribed to this mailing list so I can gain a better understanding of Histology. Thanks, Hannah Long hll0211@aol.com From kaleid11 <@t> yahoo.com Wed Jan 18 21:01:43 2006 From: kaleid11 <@t> yahoo.com (Adam Perry) Date: Wed Jan 18 21:01:51 2006 Subject: [Histonet] ICC Background In-Reply-To: <20060118190831.8ggkko488cw84swo@www.bu.edu> Message-ID: <20060119030143.9152.qmail@web30403.mail.mud.yahoo.com> Hey Amy, I'm sure this will be the first of many emails asking for clarification...but here goes: 1) How are you visualizing your antibodies and what is your "background"? Is it fluorescence and excess autofluorescence? Is it DAB/NiDAB and excessive non-specific chromogen deposition? Are you using any amplification systems (avidin-biotin-HRP, tyramine signal amplification, etc.)? 2) What antibodies are you using? What species are they raised in and what epitopes are they directed against? Have you run a titration of your primaries and secondaries...if so, what concentrations have you tried? 3) Are you using a blocking step? If so, what are you blocking with? 4) What buffer are you doing rinses/incubations in? Tris-buffered saline, phosphate-buffered saline, or other? Is there any detergent in your buffers (Triton X-100, etc.)? 5) Have you tried an incubation in sodium borohydride to reduce unreacted aldehydes (such as from your fixation) into primary alcohols? 6) How long are your antibody incubation steps? And are you doing ICC on free-floating sections or on slides? I think those are my primary questions for now... Cheers, Adam Adam Perry Department of Physiology and Biophysics University of Illinois Chicago, IL 60612 acjanes@bu.edu wrote: Hi, I really need some suggestions about the background I am getting when running immunocytochemistry. I am using mouse brain tissue that is 30microns thick and has been perfused with 4% para. We have used different primaries and secondaries and we always seem to get really high background regardless of which primary or secondary we are using. I have also gotten high background in my non-primary control. The background is so high I can't even tell if there is any specific binding. I have run ICC many times in the past and I have gotten it to work before. I am not sure what is going on so if anyone has any suggestions please let me know! Thanks! Amy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From Barry.R.Rittman <@t> uth.tmc.edu Thu Jan 19 07:27:08 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Thu Jan 19 07:28:19 2006 Subject: [Histonet] tooth curl. Long reply Message-ID: Trisha It is common for dentin especially to lift up off the slides especially at the edges. Four things that you might try 1. One trick that works is when the sections are being mounted on slides to brush them out. This is much more gentle than using needles. Flattened sections on a warming plate are allowed to get almost dry. Then use a fine camel or sable hair paint brush and gently brush the dentin of the tooth to eliminate folds or curling. Touch the paintbrush to a piece of filter paper to eliminate excess moisture that you have picked up from the section. If the dentin of the tooth in an area dries before the fold is removed then add just a touch of fluid to that area and start again. 2. Paraffin sections of bone and teeth often adhere more tenaciously to the slide if when the sections are dry to then gently heat over a small flame until the paraffin just melts. No need to BBQ them!! 3. Once slides have been deparaffinized and are in absolute alcohol, they can be coated with a thin layer of celloidin. 0.5% in alcohol: ether mixture is the usual solution. The slides are dipped in the solution for a few seconds, allowed to semi dry until a sheen appears on the surface and then placed directly into 70% ethanol. This coating of celloidin is an excellent method to keep sections of tooth stuck down. There are however a few drawbacks. Celloidin is impermeable to enzymes and so cannot be used for digestion studies. Celloidin is also stained irreversible by some stains, Celestine blue, alcian blue etc. If any of these are problems then the celloidinization can be carried out after staining. Celloidin is soluble in absolute alcohol. Therefore the final step before going into the clearing agent should be in alcohol: chloroform (absolute ethanol and 15% or more chloroform). 4. Finally. Xylene tends to lift edges of tooth sections and they often will resist flattening. Instead of xylene can use terpineol. This is also listed as "oil of lilacin". Terpineol is miscible with absolute ethanol, xylene and most mountants. It dries very slowly and tissues remain flexible. The sections are covered with terpineol, drained and excess is removed by blotting the section with bibulous paper backed with paper towels. Mountant then applied etc. Hope that this helps If you need further details please contact me. 713-500-4134 Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Trisha Emry Sent: Wednesday, January 18, 2006 6:38 PM To: histo Subject: [Histonet] tooth curl I just did serial sections through a decaled, paraffin embedded tooth. I had trouble keeping sections stained with H & E on "plus" slides and they often curled up at the edges and folded. Any advice on how to deal with the ones cut and/or future teeth? Thanks, Trisha U of Washington, Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Thu Jan 19 07:42:29 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Jan 19 07:42:37 2006 Subject: [Histonet] DIF Controls In-Reply-To: <813FB33DA405334F947F8BFC6EBD0B2A02E2EB@bbplsrv1.bbpl> Message-ID: <20060119134229.23855.qmail@web61211.mail.yahoo.com> Steve: We use to run positive controls for each antibody; this is the only way that you can be sure that your dilution worked. As positive controls we run positive cases with particularly high positive signal for each FITC conjugated antibody. If you don't do that you could end repporting a false negative since you don't have the + control to compare with. Hope this helps! Ren? J. Steve Weaver wrote: **New** CAP Question: ANP.21850 "Are appropriate positive and negative controls performed with each case tested using immunofluorescence?" We currently just run one (+) control, usually IgG or C3, for the entire run that includes IgA, IgG, IgM, C3, C4 and fibrinogen. My question is - should I be running (+) controls for all antibodies? Your thoughts please. Please indicate if you have been inspected on this new question. Thank you. sw _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. From rjbuesa <@t> yahoo.com Thu Jan 19 07:50:49 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Jan 19 07:50:56 2006 Subject: [Histonet] tooth curl In-Reply-To: Message-ID: <20060119135049.76466.qmail@web61219.mail.yahoo.com> Trisha: Try adding 1 mL of Elmer's white glue in your regular 2-liter capacity round water bath which will be equivalent to a 0.05% solution, approx. You could even add some weight over the back of the slide with the section over a flat plastic surface. You should cover the plastic surface with paraffin oil (= mineral oil) to avoid the section to adhere to the plastic. As weight you could use some lead "sinkers", the ones used to sink the fishing lines. Ren? J. Trisha Emry wrote: I just did serial sections through a decaled, paraffin embedded tooth. I had trouble keeping sections stained with H & E on "plus" slides and they often curled up at the edges and folded. Any advice on how to deal with the ones cut and/or future teeth? Thanks, Trisha U of Washington, Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos ? Showcase holiday pictures in hardcover Photo Books. You design it and we?ll bind it! From TillRenee <@t> uams.edu Thu Jan 19 08:39:04 2006 From: TillRenee <@t> uams.edu (Till, Renee) Date: Thu Jan 19 08:39:34 2006 Subject: [Histonet] antigen retrieval on cryosections Message-ID: Any helpful hints for keeping cryosections on the slide during antigen retrieval? I am not very experience in cryosections used for immunos, but they have cd markers that have to be antigen retrieved. Using the same retrieval as I do however often makes the sections come off the slides. I believe they use just Superfrost Plus slides. Would any other type of slide help? Right now they use Citra Plus in the microwave for 2 minutes on power 10, then 10 minutes on power 1. That is what I do for my paraffin sections and it is fine. Perhaps less time? Or even a different antigen retrieval solution? I have asked for help with problems before that this lab has had. They always hire techs for everthing else and want me to teach them the histo part of it. ================================================================================================ 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 sjchtascp <@t> yahoo.com Thu Jan 19 08:40:51 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Thu Jan 19 08:41:06 2006 Subject: [Histonet] air drying-white sections?? Message-ID: <20060119144051.37842.qmail@web90208.mail.scd.yahoo.com> I've noticed that when I air dry my sections before drying them some of them are turning a chauky white. Might that indicate anything important. I really have not noticed this before in other places I work. I wipe off as much excess water with a kimwipe prior to letting the sections air dry vertically. Any thoughts. Steve --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. From rjbuesa <@t> yahoo.com Thu Jan 19 08:51:21 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Jan 19 08:51:29 2006 Subject: [Histonet] antigen retrieval on cryosections In-Reply-To: Message-ID: <20060119145121.31560.qmail@web61212.mail.yahoo.com> Ren?e: Antigen retrieval or HIER, as you well know, was developed as a technique to compensate for the crosslinkage produced by formalin when fixing tissues. If you are using cryosections on unfixed tissue, you just don't need to do HIER. In the same way that you will not need HIER if using tissues fixed with alcoholic fixatives that will fixed proteins by precipitation and not by crosslinking. Summary; you don't need HIER in frozen sections, I never used it and went directly to the IHC protocol. Hope this will help you! Ren? J. "Till, Renee" wrote: Any helpful hints for keeping cryosections on the slide during antigen retrieval? I am not very experience in cryosections used for immunos, but they have cd markers that have to be antigen retrieved. Using the same retrieval as I do however often makes the sections come off the slides. I believe they use just Superfrost Plus slides. Would any other type of slide help? Right now they use Citra Plus in the microwave for 2 minutes on power 10, then 10 minutes on power 1. That is what I do for my paraffin sections and it is fine. Perhaps less time? Or even a different antigen retrieval solution? I have asked for help with problems before that this lab has had. They always hire techs for everthing else and want me to teach them the histo part of it. ================================================================================================ 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 --------------------------------- Yahoo! Photos ? Showcase holiday pictures in hardcover Photo Books. You design it and we?ll bind it! From dobbin <@t> upei.ca Thu Jan 19 08:56:35 2006 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Thu Jan 19 09:01:28 2006 Subject: [Histonet] antigen retrieval on cryosections In-Reply-To: Message-ID: <43CF7063.16858.7B53D7@acad1.cs.upei.ca> Reneee, If your tissues are not formalin (or aldehyde) fixed, then antigen retrieval is not required. Greg Date sent: Thu, 19 Jan 2006 08:39:04 -0600 From: "Till, Renee" To: histonet@lists.utsouthwestern.edu Subject: [Histonet] antigen retrieval on cryosections > Any helpful hints for keeping cryosections on the slide during antigen > retrieval? I am not very experience in cryosections used for immunos, > but they have cd markers that have to be antigen retrieved. Using the > same retrieval as I do however often makes the sections come off the > slides. I believe they use just Superfrost Plus slides. Would any > other type of slide help? Right now they use Citra Plus in the > microwave for 2 minutes on power 10, then 10 minutes on power 1. That > is what I do for my paraffin sections and it is fine. Perhaps less > time? Or even a different antigen retrieval solution? I have asked for > help with problems before that this lab has had. They always hire > techs for everthing else and want me to teach them the histo part of > it. > > > > > ====================================================================== > ========================== > > Confidentiality Notice: This e-mail message, including any > attachments, is for the sole use of the intended recipient(s) and may > contain confidential and privileged information. Any unauthorized > review, use, disclosure or distribution is prohibited. If you are not > the intended recipient, please contact the sender by reply e-mail and > destroy all copies of the original message. > ====================================================================== > ========================== > _______________________________________________ Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Thu Jan 19 09:03:34 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Jan 19 09:04:02 2006 Subject: [Histonet] DIF Controls Message-ID: Yes, I would and do. Robyn OHSU From rjbuesa <@t> yahoo.com Thu Jan 19 09:09:01 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Jan 19 09:09:10 2006 Subject: [Histonet] air drying-white sections?? In-Reply-To: <20060119144051.37842.qmail@web90208.mail.scd.yahoo.com> Message-ID: <20060119150901.97646.qmail@web61214.mail.yahoo.com> Steve: "Chauky white" sections are often an indication of poorly dehydrated tissue before the antemedium/infiltration step. How is the microscopic detail in those section? Do you have this problem in all of your sections? Are they more frequent in a given type of tissue? Answering to these questions could lead you to the cause of the problem. Hope this will help you. Ren? J. Steven Coakley wrote: I've noticed that when I air dry my sections before drying them some of them are turning a chauky white. Might that indicate anything important. I really have not noticed this before in other places I work. I wipe off as much excess water with a kimwipe prior to letting the sections air dry vertically. Any thoughts. Steve --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. From gcallis <@t> montana.edu Thu Jan 19 09:32:20 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Jan 19 09:32:35 2006 Subject: Fwd: Re: [Histonet] antigen retrieval on cryosections Message-ID: <6.0.0.22.1.20060119083058.01b4ca00@gemini.msu.montana.edu> >There is a recent interesting and excellent publication on this subject > >Shuji Yamashita and Yasunori Okada >Application of Heat-induced Antigen Retrieval to Aldehyde-fixed Fresh >Frozen Sections >J. Histochem. Cytochem., Nov 2005; 53: 1421 - 1432. > > >Erie Scientific makes a new plus charge slide, called EXCEL (sp?) Have >them send a sample to try as these are supposed to be more resistant to >retrieval, mainly EDTA. Worth a try to see it they are better for frozen >sections. If you do NOT use NBF or aldehyde fixation for a frozen section >in the first place, then retrieval is NOT needed. You can do acetone >fixation for CD markers and live retrieval free. > >We never do retrieval on fresh tissue frozen sections but we do NOT use >PFA or NBF for fixation which mess up too many of our CD markers. > >. At 07:39 AM 1/19/2006, you wrote: >>Any helpful hints for keeping cryosections on the slide during antigen >>retrieval? I am not very experience in cryosections used for immunos, >>but they have cd markers that have to be antigen retrieved. Using the >>same retrieval as I do however often makes the sections come off the >>slides. I believe they use just Superfrost Plus slides. Would any other >>type of slide help? Right now they use Citra Plus in the microwave for 2 >>minutes on power 10, then 10 minutes on power 1. That is what I do for >>my paraffin sections and it is fine. Perhaps less time? Or even a >>different antigen retrieval solution? I have asked for help with >>problems before that this lab has had. They always hire techs for >>everthing else and want me to teach them the histo part of it. >> >> Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From BennettW <@t> pac.dfo-mpo.gc.ca Thu Jan 19 11:24:36 2006 From: BennettW <@t> pac.dfo-mpo.gc.ca (BennettW@pac.dfo-mpo.gc.ca) Date: Thu Jan 19 11:21:26 2006 Subject: [Histonet] Invertebrates in Sediment Message-ID: <7CBBD627E4E688499349A5D11D078316031309DB@msgpacpbs.pac.dfo-mpo.ca> Hello Histonetters, Thanks in advance for spending any time with this query! I have a researcher that has collected sediment samples, preserved with NBF, and wants to stain any invertebrates in the sediment so that they can be enumerated and identified. The researcher wants to use rose bengal but doesn't know at what concentration and neither do I. Can anyone help? Thanks Bill Bennett Histologist Fisheries and Oceans Canada Pacific Biological Station From rjbuesa <@t> yahoo.com Thu Jan 19 11:51:11 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Jan 19 11:51:21 2006 Subject: [Histonet] Invertebrates in Sediment In-Reply-To: <7CBBD627E4E688499349A5D11D078316031309DB@msgpacpbs.pac.dfo-mpo.ca> Message-ID: <20060119175111.53298.qmail@web61214.mail.yahoo.com> Bill: "Invertebrate" is an extraordinarily encompassing zoological term and as such the techniques that can be used to "stain" its components are almost as numerous. Rose bengal has been used in many procedures one of which is to stain soil bacteria. Conn (1928) used the following recipe: Dist.water→100 mL + hydrated phenol→ 5 mL + calcium chloride → 10 mg + rose bengal → 1g. He used smears of gelatin suspended soil dried at 100?C stained on the slide and heated in a water bath x 1 min→ wash→dry. I don't think that your researcher wants to do this though. It seems more likely that what the researcher wants is to stain and count while suspended in liquid. If this is the case the technique will depend on the organisms been looked for. Perhaps you would be better off by using iodine solution that will permit the visualization of many an invertebrate. Mix at a 50:50 rate Lugol's solution with a saturated aqueous solution of Eoin Y Mix 1 part of the solution with 10 parts of the sample. I hope this will help. Ren? J. BennettW@pac.dfo-mpo.gc.ca wrote: Hello Histonetters, Thanks in advance for spending any time with this query! I have a researcher that has collected sediment samples, preserved with NBF, and wants to stain any invertebrates in the sediment so that they can be enumerated and identified. The researcher wants to use rose bengal but doesn't know at what concentration and neither do I. Can anyone help? Thanks Bill Bennett Histologist Fisheries and Oceans Canada Pacific Biological Station _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. From ROrr <@t> enh.org Thu Jan 19 12:26:04 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Thu Jan 19 12:26:13 2006 Subject: [Histonet] Slide storage Message-ID: Hello everyone, I need to store unstained slides for what might be 6 months to a year. These slides may be used for some IHC staining. I do not know what anibodies, but I would imagine they are not the ones used in a routine clinical lab setting. I have some options, for storage, please give opinions. Store the slides in a slide mailer, then wrap in foil and place in -80 freezer? Or one of my pathologists talked about coating each unstained slide in a layer of paraffin. Has anyone done this technique? I would assume that I just cut, dry, then melt the slide, then dip the slide in paraffin? Many thanks for your help. Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 From bjdewe <@t> aol.com Thu Jan 19 12:28:59 2006 From: bjdewe <@t> aol.com (bjdewe@aol.com) Date: Thu Jan 19 12:29:20 2006 Subject: [Histonet] Counter stain Message-ID: <8C7EB3D595FDB23-F0C-1832@MBLK-M42.sysops.aol.com> I am doing a peroxidase DAB stain for an antigen that is present only in small quantities in the nucleus. I am doing it on rat bone. If I counterstain with hematoxylin it covers the DAB stain. What else can I use that will only stain the cytoplasm and not the nucleus. i thought of methyl green but it too is supposed to stain the nucleus. I thought of neutral red but it won't look so good with the brown. Suggestions? Cheers, Lorie ******************************************************************* 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 bhewlett <@t> cogeco.ca Thu Jan 19 12:30:00 2006 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Thu Jan 19 12:30:14 2006 Subject: Fw: [Histonet] Invertebrates in Sediment Message-ID: <003a01c61d26$5b5ea590$6400a8c0@mainbox> ----- Original Message ----- From: "Bryan Hewlett" To: Sent: Thursday, January 19, 2006 1:14 PM Subject: Re: [Histonet] Invertebrates in Sediment > Bill, > > The third edition of "Staining Procedures used by the Biological Stain > Commission" edited by George Clark, > gives a procedure (Walton 1952) for staining foraminiferans alive at the > time of fixation of marine sediments "without incidental staining of > organic or inorganic debris". > > The concentration of Rose Bengal is given as 0.1% aqueous and the staining > time as 10 minutes. > There is a footnote that the concentration of dye is not critical. > Since the dye is also a fluorochrome, enumeration may be easier at low > power by fluorescence microscopy. > > Bryan > > ----- Original Message ----- > From: > To: > Sent: Thursday, January 19, 2006 12:24 PM > Subject: [Histonet] Invertebrates in Sediment > > >> Hello Histonetters, >> >> Thanks in advance for spending any time with this query! I have a >> researcher that has collected sediment samples, preserved with NBF, and >> wants to stain any invertebrates in the sediment so that they can be >> enumerated and identified. The researcher wants to use rose bengal but >> doesn't know at what concentration and neither do I. Can anyone help? >> >> Thanks >> Bill Bennett >> Histologist >> Fisheries and Oceans Canada >> Pacific Biological Station >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > From sharon.osborn <@t> dnax.org Thu Jan 19 12:36:54 2006 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Thu Jan 19 12:37:22 2006 Subject: [Histonet] IHC slides and sections Message-ID: <29B25753F6B1D51196110002A589D444032C49B1@PALMSG30.us.schp.com> Thanks a bunch for all the wonderful suggestions. We tried several of the suggestions and sections still came off. The charged slides were still 'in date'. We switched to a different slide and the sections remaned on. The charged slides have different "stickiness" depending upon the vendor. In the past we have used some slides that were 4-5 years old and the stickiness was still present-excellent charge retention I guess. However, we now have solved the problem with a stickier slide on the charge. Your suggestions are helpful for our use overall and several are being implement as a regular protocol. Have a great weekend! And, again THANKS! Sharon Osborn DNAX, SP BioPharma Palo Alto, CA 650-496-6539 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From PMonfils <@t> Lifespan.org Thu Jan 19 12:37:28 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Thu Jan 19 12:37:39 2006 Subject: [Histonet] Invertebrates in Sediment Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171764B@lsexch.lsmaster.lifespan.org> Rose bengal is widely used in sediment studies in both fresh and salt water because it brightly stains just about any organic matter without staining inorganic matter. The exact concentration isn't critical. One common method is to dissolve 5 grams of rose bengal in 100 ml commercial formaldehyde solution saturated with sodium borate. This is easy to carry in the field, and is used by adding one part of this solution to 9 parts of sediment sample, which results in a 10% buffered formalin solution containing 0.5 % rose bengal. Of course the sample can simply be fixed in the field, and the dye added after returning to the lab. The following sites may help: http://www.eman-rese.ca/eman/ecotools/protocols/freshwater/benthics/sample.h tml http://www.nefsc.noaa.gov/nefsc/publications/tm/tm167/tm167p6.htm From brett_connolly <@t> merck.com Thu Jan 19 12:40:32 2006 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Thu Jan 19 12:41:06 2006 Subject: [Histonet] Counter stain Message-ID: <355C35514FEAC9458F75947F5270974D67CCA3@usctmx1103.merck.com> Lorie, I've used 0.25% Metanil Yellow and/or Light Green...either one works fine, just play with the times to get the staining intensity you want. Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of bjdewe@aol.com Sent: Thursday, January 19, 2006 1:29 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Counter stain I am doing a peroxidase DAB stain for an antigen that is present only in small quantities in the nucleus. I am doing it on rat bone. If I counterstain with hematoxylin it covers the DAB stain. What else can I use that will only stain the cytoplasm and not the nucleus. i thought of methyl green but it too is supposed to stain the nucleus. I thought of neutral red but it won't look so good with the brown. Suggestions? Cheers, Lorie ******************************************************************* 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 ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From PMonfils <@t> Lifespan.org Thu Jan 19 12:43:19 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Thu Jan 19 12:43:33 2006 Subject: [Histonet] Counter stain Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171764C@lsexch.lsmaster.lifespan.org> I routinely use 1% methyl green for 1 minute as a counterstain for peroxidase methods, with both antibodies and lectins. It does stain the nucleus, but much less densely then hematoxylin does - just about darkly enough to show where the nuclei are. Peroxidase stains show up clearly against it. From anh2006 <@t> med.cornell.edu Thu Jan 19 12:44:47 2006 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Thu Jan 19 12:44:53 2006 Subject: [Histonet] Counter stain In-Reply-To: <8C7EB3D595FDB23-F0C-1832@MBLK-M42.sysops.aol.com> References: <8C7EB3D595FDB23-F0C-1832@MBLK-M42.sysops.aol.com> Message-ID: "Light Green" from Biomeda (can be ordered through VWR). It is a light green cytoplasmic stain, very nice actually!! >I am doing a peroxidase DAB stain for an antigen that is present >only in small quantities in the nucleus. I am doing it on rat bone. >If I counterstain with hematoxylin it covers the DAB stain. What >else can I use that will only stain the cytoplasm and not the >nucleus. i thought of methyl green but it too is supposed to stain >the nucleus. I thought of neutral red but it won't look so good with >the brown. Suggestions? > >Cheers, >Lorie > >******************************************************************* >Confidentiality Notice: This e-mail message, including any >attachments, is for the sole use of the intended recipient(s)and may >contain confidential and privileged information. Any unauthorized >review, use, disclosure or distribution is prohibited. >If you are not the intended recipient, please contact the sender by >reply e-mail and destroy all copies of the original message. >******************************************************************* >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- From BMolinari <@t> heart.thi.tmc.edu Thu Jan 19 12:49:52 2006 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Thu Jan 19 12:55:31 2006 Subject: [Histonet] Z fix Message-ID: Hi, I have some mouse hearts in Z Fix. I am going to be out of town for the next week, can they stay in Z Fix or should I transfer them to 70%? Thanks. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) From rjbuesa <@t> yahoo.com Thu Jan 19 12:55:34 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Jan 19 12:55:42 2006 Subject: [Histonet] Slide storage In-Reply-To: Message-ID: <20060119185534.18570.qmail@web61225.mail.yahoo.com> Rebecca: Try to isolate the slides as much from air as you can. Inside a plastic box (those that can contain 25 slides) sealed in a pouch with as less air as possible and in a refrigerator will be the best option. In a paper I wrote (JOH 28(2):89-97, 2005) I was able to demonstrate that "normal" storage of slides in the lab. environment for more than 6 weeks reduces epitope reactivity in a statistically significant way. Ren? J. "Orr, Rebecca" wrote: Hello everyone, I need to store unstained slides for what might be 6 months to a year. These slides may be used for some IHC staining. I do not know what anibodies, but I would imagine they are not the ones used in a routine clinical lab setting. I have some options, for storage, please give opinions. Store the slides in a slide mailer, then wrap in foil and place in -80 freezer? Or one of my pathologists talked about coating each unstained slide in a layer of paraffin. Has anyone done this technique? I would assume that I just cut, dry, then melt the slide, then dip the slide in paraffin? Many thanks for your help. Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos ? Showcase holiday pictures in hardcover Photo Books. You design it and we?ll bind it! From juan.gutierrez <@t> christushealth.org Thu Jan 19 13:07:36 2006 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Thu Jan 19 13:07:49 2006 Subject: [Histonet] coverslipper Message-ID: Richard-Allan Sci. is supposed to have some new glass coverslips that are "pre-glued". All you have to do is activate them with xylene or some other clearing solution. We are waiting to see if they can be used in our coverslipper without much trouble in doing the switch. Hope this is what you were looking for. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Wednesday, January 18, 2006 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] coverslipper When I was NSH this past year, there was a coverslipper there that had the mounting media directly on the coverslips and a solution activated it to mount it to the slide. I cannot remember the company who made this. If you know would you please email me or if you are the company who makes this coverslipper contact me please. Thanks. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org ------------------------------------------------------------------------ ------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ======================================================================== ====== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sbreeden <@t> nmda.nmsu.edu Thu Jan 19 13:14:26 2006 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Thu Jan 19 13:14:36 2006 Subject: [Histonet] Sub-X and Coverslipper Tape Message-ID: I'm considering a change from my current xylene/Citrisolv setup to totally Sub-X. I know that Sub-X is approved for my Leica ASP300 and I know I can use it in my staining setup, but before I make this major commitment, I'd like to know if anyone has experienced any incompatibility issues between Sub-X and the coverslipping tape. I love my tape coverslipper and have had no problems with lifting, etc., but as they say in Texas..."If I ain't broke, don't fix it". Thank you, in advance. May the Force be with you. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From tpmorken <@t> labvision.com Thu Jan 19 13:24:56 2006 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Thu Jan 19 13:25:10 2006 Subject: [Histonet] Slide storage Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D557@usca0082k08.labvision.apogent.com> Rebecca, you might be interested in this study on tissue array longevity. Lab Invest. 2004 Aug;84(8):1071-8. Long-term preservation of antigenicity on tissue microarrays. DiVito KA, Charette LA, Rimm DL, Camp RL. Department of Pathology, Yale University, New Haven, CT, USA. Tissue microarrays have facilitated the evaluation of large cohort studies; however, there is little data on the best method for preserving sections once they are cut. We assessed three methods of storing precut breast cancer microarray slides: paraffin coating and storage in a nitrogen desiccator, either alone or in combination. We tested the durability of three antigens, cytokeratin, estrogen receptor, and Ki-67 on microarrays stored under these conditions for 3 months at room temperature. Staining was assessed with both manual scoring using traditional brown stain (0-3+) as well as automated scoring using fluorescently stained sections. Staining intensity was compared to that obtained from freshly cut slides. Slides stored under ambient conditions (room temperature and air) for 3 months exhibited marked degradation of all target antigens, in some cases resulting in slides that were virtually unreadable. We found that combined paraffin coating and nitrogen storage resulted in the best preservation of antigenicity, with retention of 72-99% of the antigenicity of a freshly cut slide, depending upon the marker and detection system used. The use of either paraffin coating or nitrogen storage alone protected slides to a lesser degree. PMID: 15195116 [PubMed - indexed for MEDLINE] Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Thursday, January 19, 2006 10:56 AM To: Orr, Rebecca; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Slide storage Rebecca: Try to isolate the slides as much from air as you can. Inside a plastic box (those that can contain 25 slides) sealed in a pouch with as less air as possible and in a refrigerator will be the best option. In a paper I wrote (JOH 28(2):89-97, 2005) I was able to demonstrate that "normal" storage of slides in the lab. environment for more than 6 weeks reduces epitope reactivity in a statistically significant way. Ren? J. "Orr, Rebecca" wrote: Hello everyone, I need to store unstained slides for what might be 6 months to a year. These slides may be used for some IHC staining. I do not know what anibodies, but I would imagine they are not the ones used in a routine clinical lab setting. I have some options, for storage, please give opinions. Store the slides in a slide mailer, then wrap in foil and place in -80 freezer? Or one of my pathologists talked about coating each unstained slide in a layer of paraffin. Has anyone done this technique? I would assume that I just cut, dry, then melt the slide, then dip the slide in paraffin? Many thanks for your help. Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos - Showcase holiday pictures in hardcover Photo Books. You design it and we'll bind it! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> cdc.gov Thu Jan 19 13:26:46 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Jan 19 13:45:11 2006 Subject: [Histonet] coverslipper Message-ID: So it's the coverslips that are being marketed? I guess if they don't stick and can be picked up by other machines and then have xylene instead of mountant dropped onto the slide it might work. Interesting........... Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 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 GUTIERREZ, JUAN Sent: Thursday, January 19, 2006 2:08 PM To: Horn, Hazel V; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] coverslipper Richard-Allan Sci. is supposed to have some new glass coverslips that are "pre-glued". All you have to do is activate them with xylene or some other clearing solution. We are waiting to see if they can be used in our coverslipper without much trouble in doing the switch. Hope this is what you were looking for. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Wednesday, January 18, 2006 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] coverslipper When I was NSH this past year, there was a coverslipper there that had the mounting media directly on the coverslips and a solution activated it to mount it to the slide. I cannot remember the company who made this. If you know would you please email me or if you are the company who makes this coverslipper contact me please. Thanks. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org ------------------------------------------------------------------------ ------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ======================================================================== ====== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GoodwinD <@t> pahosp.com Thu Jan 19 13:51:44 2006 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Thu Jan 19 13:51:54 2006 Subject: [Histonet] Histology workflow/process analysis Message-ID: <80CDD9C3FEEAFD4982B114C4A6DFD00EE4A2E6@uphsmbx2.UPHS.PENNHEALTH.PRV> Greetings, Histonet. Can anyone recommend a program or procedure to track and document workflow in the Histology lab, the goal being process improvement? Thanks! Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Preston 655-C ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com From bjdewe <@t> aol.com Thu Jan 19 14:04:12 2006 From: bjdewe <@t> aol.com (bjdewe@aol.com) Date: Thu Jan 19 14:04:31 2006 Subject: [Histonet] IHC slides and sections In-Reply-To: <29B25753F6B1D51196110002A589D444032C49B1@PALMSG30.us.schp.com> References: <29B25753F6B1D51196110002A589D444032C49B1@PALMSG30.us.schp.com> Message-ID: <8C7EB4AA681C7C3-F0C-22C7@MBLK-M42.sysops.aol.com> So what slides are the most adhesive? I only knew about the Superfrost + slides. Are there more brands?? Lorie ******************************************************************* 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. ******************************************************************* -----Original Message----- From: Osborn, Sharon To: 'histonet@lists.utsouthwestern.edu' Sent: Thu, 19 Jan 2006 13:36:54 -0500 Subject: [Histonet] IHC slides and sections Thanks a bunch for all the wonderful suggestions. We tried several of the suggestions and sections still came off. The charged slides were still 'in date'. We switched to a different slide and the sections remaned on. The charged slides have different "stickiness" depending upon the vendor. In the past we have used some slides that were 4-5 years old and the stickiness was still present-excellent charge retention I guess. However, we now have solved the problem with a stickier slide on the charge. Your suggestions are helpful for our use overall and several are being implement as a regular protocol. Have a great weekend! And, again THANKS! Sharon Osborn DNAX, SP BioPharma Palo Alto, CA 650-496-6539 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Thu Jan 19 14:20:53 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Thu Jan 19 14:21:04 2006 Subject: [Histonet] Counter stain - P.S. Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171764E@lsexch.lsmaster.lifespan.org> P.S. Dyes like light green or fast green, which stain everything the same shade and don't differentiate nuclei, are suitable as counterstains for some immuno work. However, in staining for a nuclear antigen it is particularly important to have the nuclei visible. Otherwise how can you be sure that the staining you see is actually in the nuclei? That's the advantage of methyl green. It doesn't stain anything very dark, but it does stain the nuclei a little darker than the cytoplasm, so you can see where they are. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > bjdewe@aol.com > Sent: Thursday, January 19, 2006 10:28 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Counter stain > > I am doing a peroxidase DAB stain for an antigen that is present only in > small quantities in the nucleus. I am doing it on rat bone. If I > counterstain with hematoxylin it covers the DAB stain. What else can I use > that will only stain the cytoplasm and not the nucleus. i thought of > methyl green but it too is supposed to stain the nucleus. I thought of > neutral red but it won't look so good with the brown. Suggestions? > > Cheers, > Lorie > > ******************************************************************* > Confidentiality Notice: This e-mail message, including any attachments, is > for the sole use of the intended recipient(s)and may contain confidential > and privileged information. Any unauthorized review, use, disclosure or > distribution is prohibited. > If you are not the intended recipient, please contact the sender by reply > e-mail and destroy all copies of the original message. > ******************************************************************* > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From naje1972 <@t> yahoo.com Thu Jan 19 14:35:24 2006 From: naje1972 <@t> yahoo.com (cynthia haynes) Date: Thu Jan 19 14:35:31 2006 Subject: [Histonet] Prill Message-ID: <20060119203524.96432.qmail@web33014.mail.mud.yahoo.com> Hello everyone in Histoland. A few days ago there was a discusion about prill and how powdery it is and how it gets all the place whenever you are trying to weight the amount you need. Some one mentioned Fisher had a brand that was better and easier to weight out. If you would contact me with this information I would appreciate greatly. My e-mail address is naje1972@yahoo.com. My name is Cynthia Haynes. Thanks in advance. From akbitting <@t> geisinger.edu Thu Jan 19 14:38:44 2006 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Thu Jan 19 14:39:29 2006 Subject: [Histonet] Cat Hematoxylin Message-ID: I'm looking at a double-staining protocol that uses Cat Hematoxylin. Does anyone know what Cat Hematoxylin is? I don't work in research, so maybe this is something new or more widely utilized in a research setting. I have a feeling that I'm going to feel really stupid when I hear the answer, but I can't let this go. I need to know. Thanks, in advance, for enlightening me. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From akbitting <@t> geisinger.edu Thu Jan 19 14:45:00 2006 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Thu Jan 19 14:45:33 2006 Subject: [Histonet] acetylcholinesterase staining Message-ID: Our track record here with our acetylcholinesterase stain is about 50/50. We make up all the solutions fresh on Monday and it works fine, by Tuesday ...nothing. Can anyone send me a good reliable Acetylcholinesterase staining method? Maybe someone elses will work better for us. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From HornHV <@t> archildrens.org Thu Jan 19 15:50:02 2006 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Thu Jan 19 15:49:39 2006 Subject: [Histonet] coverslipper Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDF61@EMAIL.archildrens.org> Actually it's a whole new coverslipper. I suppose you might could use the coverslips in another machine with their activator. But my guess is probably not. If that was the case they would have just made the coverglass? Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail 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: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Thursday, January 19, 2006 1:27 PM To: GUTIERREZ, JUAN; Horn, Hazel V; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] coverslipper So it's the coverslips that are being marketed? I guess if they don't stick and can be picked up by other machines and then have xylene instead of mountant dropped onto the slide it might work. Interesting........... Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 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 GUTIERREZ, JUAN Sent: Thursday, January 19, 2006 2:08 PM To: Horn, Hazel V; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] coverslipper Richard-Allan Sci. is supposed to have some new glass coverslips that are "pre-glued". All you have to do is activate them with xylene or some other clearing solution. We are waiting to see if they can be used in our coverslipper without much trouble in doing the switch. Hope this is what you were looking for. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Wednesday, January 18, 2006 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] coverslipper When I was NSH this past year, there was a coverslipper there that had the mounting media directly on the coverslips and a solution activated it to mount it to the slide. I cannot remember the company who made this. If you know would you please email me or if you are the company who makes this coverslipper contact me please. Thanks. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org ------------------------------------------------------------------------ ------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ======================================================================== ====== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From anh2006 <@t> med.cornell.edu Thu Jan 19 15:52:54 2006 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Thu Jan 19 15:53:02 2006 Subject: [Histonet] Counter stain - P.S. In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE0171764E@lsexch.lsmaster.lifespan.org> References: <09C945920A6B654199F7A58A1D7D1FDE0171764E@lsexch.lsmaster.lifespan.org> Message-ID: Good point. I personally use hematoxylin for many of my weak nuclear stains especially when quantitating the stain. I just stain them very lightly. >P.S. Dyes like light green or fast green, which stain everything the same >shade and don't differentiate nuclei, are suitable as counterstains for some >immuno work. However, in staining for a nuclear antigen it is particularly >important to have the nuclei visible. Otherwise how can you be sure that >the staining you see is actually in the nuclei? That's the advantage of >methyl green. It doesn't stain anything very dark, but it does stain the >nuclei a little darker than the cytoplasm, so you can see where they are. -- From PMonfils <@t> Lifespan.org Thu Jan 19 16:20:07 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Thu Jan 19 16:20:17 2006 Subject: [Histonet] Cat Hematoxylin Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171764F@lsexch.lsmaster.lifespan.org> Never heard of it! I just had to do a google search on this one! I found a few places that sell it, but no really complete description of exactly what it is. Here's the best writeup I could find: http://www.biocare.net/SpecialStains.htm From jlinda <@t> ces.clemson.edu Thu Jan 19 17:08:43 2006 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Thu Jan 19 17:06:22 2006 Subject: [Histonet] Region III Meeting Message-ID: <5.2.1.1.2.20060119175008.01f38150@mailhost.ces.clemson.edu> Region III Meeting March 10 -12, 2006 CHARLESTON AWAITS! The wonderful city of Charleston, South Carolina, and the South Carolina Society of Histotechnology are playing host to the 2006 Region III meeting. The meeting is being held at the beautiful Charleston Harbor Resort & Marina located on Mount Pleasant with a direct view of the harbor and Fort Sumter. See http://www.charlestonharborresort.com/. Call 843-856-0028 for reservations. Ask for the special rate of $125.00/night for the Region III Histotechnology Meeting. Please note that the deadline for reservations has been extended until January 27th. Registration starts Thursday afternoon, March 9, and in the evening we will have a two-hour vendor social with a cash bar and light finger foods. Workshops start early, at 7:30 am on Friday morning. After a full day of learning, it is time to play! We will board the Palmetto at the marina Friday afternoon around 4:30pm. We will cruise through the harbor accompanied by beer/wine and light hors d'oeuvre, ultimately arriving at an uninhabited island near the Morris Island Lighthouse. Accompanied by a little beach music, we will then indulge in a true Lowcountry experience, Frogmore Stew (does NOT contain frogs)! After this unforgettable beach experience, we will time our return with the sun's setting and enjoy the breathtaking views of Charleston's sunset-silhouetted steeples. Saturday morning, we resume our excellent line-up of workshops. Saturday evening is on your own. Water taxis leave hourly from the marina to ferry across the harbor to Charleston. The USS Yorktown is docked right outside the door, and a championship 18-hole golf course is out the other door. Come join us for lots of learning and fun! Contact Debbie Ellenburg at DEllenburg2@stfrancishealth.org for a PDF program and registration information. Fax: 864-255-1664 Phone: 864-255-1582 Cell: 864-918-6272 The schedule of workshops is as follows: Friday, March 10, 2006 7:30am -11:00am WS # 1 - Simply Silver Wanda Jones/ Paul Billings WS # 2 - eMicrotome.com a hands-on microtomy workshop Gayle Callis WS # 3 - Is Your Tissue Processor Fighting You? Fight Back. Pam Marcum Friday, March 10, 2006 12:30 pm - 4:00pm WS # 4 - Building an Immunohistochemistry Section in Histology Ethel Macrea WS # 5 - Conflict Management, the Courage to Confront Roberta Smith WS # 6 - The SWEET Workshop-Smart Working Environment Ergonomics Training Jan Minshew Saturday, March 11, 2006 8:00am -11:30 am WS # 7 - Preparing for a CAP inspection Hector Hernandez Jr. WS # 8 - Unlocking the Secrets of MOHS Grossing and Cryosectioning Mequita Praet WS # 9 - To Publish or Not to Publish? Tim and Karen Burg Saturday, March 11, 2006 1:00pm - 4:30pm WS # 10 - Decalcification: Solutions and Methods Diane Sterchi & Gayle Callis WS # 11 - The Study and Preparation of Antique Microslides/Sliders Lamar Jones WS # 12 - What Every Histotech Should Know About Water Ethel Macrea Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo.htm From jnocito <@t> satx.rr.com Thu Jan 19 18:51:57 2006 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Thu Jan 19 18:52:11 2006 Subject: [Histonet] Cat Hematoxylin References: Message-ID: <001f01c61d5b$b7672ee0$e8bd0b43@yourxhtr8hvc4p> I think Biocare Medical sells CAT hematoxylin. It's an abbreviation for something, I forget. Then again, I've been forgetting a lot of things lately. What were talking about? Oh, yeah, stop sniffing the xylene. Has to be the xylene. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Angela Bitting" To: Sent: Thursday, January 19, 2006 2:38 PM Subject: [Histonet] Cat Hematoxylin I'm looking at a double-staining protocol that uses Cat Hematoxylin. Does anyone know what Cat Hematoxylin is? I don't work in research, so maybe this is something new or more widely utilized in a research setting. I have a feeling that I'm going to feel really stupid when I hear the answer, but I can't let this go. I need to know. Thanks, in advance, for enlightening me. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.1.375 / Virus Database: 267.14.20/234 - Release Date: 1/18/2006 From jennifer.l.hofecker <@t> Vanderbilt.Edu Thu Jan 19 21:36:54 2006 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Thu Jan 19 21:41:22 2006 Subject: [Histonet] Cat Hematoxylin References: <001f01c61d5b$b7672ee0$e8bd0b43@yourxhtr8hvc4p> Message-ID: <898D946569A27444B65667A49C074052852B5F@mailbe06.mc.vanderbilt.edu> I may be remembering this incorrectly, but isn't the CAT hematoxylin named after the two wonderful men who developed it? Chuck Churukian and David Tacha? Yes, sold by biocare... I don't remember what the difference is but I would imagine that somebody from Biocare Medical could help, or you could contact Charles or David... Joe, aren't you a little afraid of what might happen to us with out those fumes : ) Have a great weekend, Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe Nocito Sent: Thu 1/19/2006 6:51 PM To: Angela Bitting; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Cat Hematoxylin I think Biocare Medical sells CAT hematoxylin. It's an abbreviation for something, I forget. Then again, I've been forgetting a lot of things lately. What were talking about? Oh, yeah, stop sniffing the xylene. Has to be the xylene. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Angela Bitting" To: Sent: Thursday, January 19, 2006 2:38 PM Subject: [Histonet] Cat Hematoxylin I'm looking at a double-staining protocol that uses Cat Hematoxylin. Does anyone know what Cat Hematoxylin is? I don't work in research, so maybe this is something new or more widely utilized in a research setting. I have a feeling that I'm going to feel really stupid when I hear the answer, but I can't let this go. I need to know. Thanks, in advance, for enlightening me. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.1.375 / Virus Database: 267.14.20/234 - Release Date: 1/18/2006 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mike_cup <@t> telus.net Thu Jan 19 23:30:17 2006 From: mike_cup <@t> telus.net (Mike Cupello) Date: Thu Jan 19 23:30:27 2006 Subject: [Histonet] Need Some Heat! Message-ID: <001d01c61d82$994863d0$6401a8c0@HOME> Hi to all of those out in Histoland... I was hoping to get some help. I have an employee who is suffering from tendonitis, and she feels that scraping blocks before trimming is adding to her injury. I know that there is some sort of device "out there" that uses heat to melt the excess wax from the outside of the block so that it will fit into the microtome chuck. Does anyone know of a type of heat block or hot knife that could accomplish this? I am looking for something that will collect the melted wax so that we don't have a huge mess to clean up after a day's work. Thanks for your time, Mike Cupello. From marktarango <@t> earthlink.net Thu Jan 19 22:45:14 2006 From: marktarango <@t> earthlink.net (Mark Adam Tarango) Date: Fri Jan 20 00:47:48 2006 Subject: [Histonet] marker of endothelial differentiation in vitro Message-ID: <10756775.1137732314465.JavaMail.root@elwamui-karabash.atl.sa.earthlink.net> okay, for endothelial cells in vitro try using CD31. I know nothing about eosin, but CD31 definately is specific for endothelial cells. Mark Tarango Date: Mon, 16 Jan 2006 11:05:28 +0100 From: Annette Ekblond Subject: [Histonet] marker of endothelial differentiation in vitro To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Dear subscribers -does anyone know of an endothelial marker specific to endothelial cells that have differentiated into tube-like structures (in vitro) - and not proliferating endothelial cultures?? Kind regards Annette CR Humlebaek Denmark From mdean <@t> uci.edu Fri Jan 20 01:40:19 2006 From: mdean <@t> uci.edu (mason dean) Date: Fri Jan 20 01:40:33 2006 Subject: [Histonet] please remove me, thanks! Message-ID: <469494EB-9102-4DA6-A797-02496F9CAEA9@uci.edu> From jkiernan <@t> uwo.ca Fri Jan 20 01:48:02 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Jan 20 01:48:11 2006 Subject: [Histonet] acetylcholinesterase staining References: Message-ID: <43D095B2.75BDD11F@uwo.ca> The dire warning following your enquiry is discouraging! Nevertheless, I make bold to reply. I have worked in the esterase histochemistry field, especially choline esterases, since the early 1960s, when most of the principles had been established. Which method is failing to work for you half of the time? These methods are extremely reliable if you go by the book (any book). Complete substrate solutions cannot be kept and used next day; that's a rule that applies to just about all enzyme activity histochemistry. John Kiernan Anatomy, UWO London, Canada ----------------------- Angela Bitting wrote: > > Our track record here with our acetylcholinesterase stain is about 50/50. We make up all the solutions fresh on Monday and it works fine, by Tuesday ...nothing. Can anyone send me a good reliable Acetylcholinesterase staining method? Maybe someone elses will work better for us. > > IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Fri Jan 20 01:48:57 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Jan 20 01:49:07 2006 Subject: [Histonet] Counter stain References: <8C7EB3D595FDB23-F0C-1832@MBLK-M42.sysops.aol.com> Message-ID: <43D095E9.BB2F937B@uwo.ca> The best contrasting colour for DAB-brown is green. Ethyl green stains nuclei, so avoid it. (Methyl green has not been made for 25+ years. Dyes sold as "methyl green" should be ethyl green, which is better. Check with the vendor, because real methyl green won't perform properly without laborious purification by you in your lab.) For counterstaining cytoplasm but not nuclei, try fast green FCF. Instructions can be found in any textbook published since about 1940. Light green SF (pre-1940) acts similarly, but fades after a few years. John Kiernan Anatomy, UWO London, Canada ____________________________________________ bjdewe@aol.com wrote: > > I am doing a peroxidase DAB stain for an antigen that is present only in small quantities in the nucleus. I am doing it on rat bone. If I counterstain with hematoxylin it covers the DAB stain. What else can I use that will only stain the cytoplasm and not the nucleus. i thought of methyl green but it too is supposed to stain the nucleus. I thought of neutral red but it won't look so good with the brown. Suggestions? > > Cheers, > Lorie > > ******************************************************************* > Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s)and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. > If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. > ******************************************************************* > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Fri Jan 20 01:50:01 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Jan 20 01:50:10 2006 Subject: [Histonet] Invertebrates in Sediment References: <7CBBD627E4E688499349A5D11D078316031309DB@msgpacpbs.pac.dfo-mpo.ca> Message-ID: <43D09629.3D8C3B2B@uwo.ca> Rose Bengal B is an anionic, lipophilic xanthene dye, related to the eosins and erythrosins. One of its documented uses is staining foraminifera in marine sediments. Get your boss to send you to the library to check out "Conn's Biological Stains" and Clark's "Staining Procedures" for references to the original methods. If it's a specialized application, go to the references cited in these books. John Kiernan London, Canada. ______________________________________________________ BennettW@pac.dfo-mpo.gc.ca wrote: > > Hello Histonetters, > > Thanks in advance for spending any time with this query! I have a > researcher that has collected sediment samples, preserved with NBF, and > wants to stain any invertebrates in the sediment so that they can be > enumerated and identified. The researcher wants to use rose bengal but > doesn't know at what concentration and neither do I. Can anyone help? > > Thanks > Bill Bennett > Histologist > Fisheries and Oceans Canada > Pacific Biological Station > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Fri Jan 20 01:51:16 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Jan 20 01:51:26 2006 Subject: [Histonet] air drying-white sections?? References: <20060119144051.37842.qmail@web90208.mail.scd.yahoo.com> Message-ID: <43D09674.D41BE103@uwo.ca> Dear Steven Coakley, Please explain how you "air dry my sections before drying them". Also, describe exactly what you do with the kimwipe (assuming that a kimwipe is a small paper hanky). If you're working with paraffin sections, it's important to let all the water drain off the slides (vertically) before letting the temperature rise to anywhere near the softening point of the wax. Histology waxes with 58C "melting points" soften at about 45C. If you use a hotplate, it gets uncomfortably hot for an applied hand in 10-20 seconds at 45C. That's the maximum allowable temperature before making a decision to melt the wax. The film of water between the ribbon and the glass must be all gone before you even think about melting the wax. This advice is in every textbook of histotechnology (microtechnique) published since about 1880, and it has appeared often in Histonet for ?10 years. The archives at http://histosearch.com are full of wisdom in this field. John Kiernan Anatomy, UWO London, Canada ____________________ Steven Coakley wrote: > > I've noticed that when I air dry my sections before drying them some of them are turning a chauky white. Might that indicate anything important. I really have not noticed this before in other places I work. I wipe off as much excess water with a kimwipe prior to letting the sections air dry vertically. Any thoughts. > > Steve =========== From gillian.2.brown <@t> gsk.com Fri Jan 20 02:50:37 2006 From: gillian.2.brown <@t> gsk.com (gillian.2.brown@gsk.com) Date: Fri Jan 20 02:50:58 2006 Subject: [Histonet] Need Some Heat! In-Reply-To: <001d01c61d82$994863d0$6401a8c0@HOME> Message-ID: Hi Mike, we use a cheap domestic iron (set on delicates), standing in a shallow collecting dish. Gill Brown GlaxoSmithKline Medicines Research Centre, UK "Mike Cupello" Sent by: histonet-bounces@lists.utsouthwestern.edu 20-Jan-2006 05:30 To histonet@lists.utsouthwestern.edu cc Subject [Histonet] Need Some Heat! Hi to all of those out in Histoland... I was hoping to get some help. I have an employee who is suffering from tendonitis, and she feels that scraping blocks before trimming is adding to her injury. I know that there is some sort of device "out there" that uses heat to melt the excess wax from the outside of the block so that it will fit into the microtome chuck. Does anyone know of a type of heat block or hot knife that could accomplish this? I am looking for something that will collect the melted wax so that we don't have a huge mess to clean up after a day's work. Thanks for your time, Mike Cupello. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BlazekL <@t> childrensdayton.org Fri Jan 20 07:22:45 2006 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Jan 20 07:25:24 2006 Subject: [Histonet] job classification Message-ID: Does anyone have a dual classification of LIS manager and bench tech? If so is the salary higher than just a bench tech? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org From brett_connolly <@t> merck.com Fri Jan 20 08:09:24 2006 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Jan 20 08:11:33 2006 Subject: [Histonet] Lot to lot antibody validation Message-ID: <355C35514FEAC9458F75947F5270974D67CCA5@usctmx1103.merck.com> I would like to hear what procedures people are using to validate different lots of antibodies, including # of specimens you might use for the validation. Thanks, Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From ROrr <@t> enh.org Fri Jan 20 08:43:52 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Jan 20 08:44:01 2006 Subject: [Histonet] slide storage feedback Message-ID: Thanks everyone for your response! I got a lot of advice on long term slide storage sent to me privately, so just to share, I would say the general consensus was ultra low temps wrapped in foil along with a desiccant. Thanks again to the wonderful Mr. Tim Morken for sharing this article on the TMA storage and the paraffin "dip". The technique was what the Pathologist was looking for and what we will be using in our project. The Histonet is such a valuable tool, and I appreciate the support and help I have gotten from peers as well as vendors. Have the best weekend possible! I'll be spending my Saturday shoveling snow. Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 ------------------------------ Message: 21 Date: Thu, 19 Jan 2006 15:50:02 -0600 WS From portera <@t> msu.edu Fri Jan 20 08:45:41 2006 From: portera <@t> msu.edu (Amy Porter) Date: Fri Jan 20 08:46:46 2006 Subject: [Histonet] Lot to lot antibody validation References: <355C35514FEAC9458F75947F5270974D67CCA5@usctmx1103.merck.com> Message-ID: <000301c61dd0$2fd29210$8e7a0923@HistoJJ> We use a known postive control and run a system negative (no primary) with each new lot of primary that is received. We only run it one time maybe at the dilution for the old lot then slightly higher and slightly lower dilution with the already known pretreatment. Slides are then reviewed and the acceptable dilution for the new lot is recorded on an "Antibody Titration Log Sheet" signed off on by the pathologist or researcher to whom the work is being provided for. Hope this helps out... ----- Original Message ----- From: "Connolly, Brett M" To: Sent: Friday, January 20, 2006 9:09 AM Subject: [Histonet] Lot to lot antibody validation >I would like to hear what procedures people are using to validate different > lots of antibodies, including # of specimens you might use for the > validation. > > Thanks, Brett > > Brett M. Connolly, Ph.D. > Merck & Co., Inc. > MRL, Imaging Research > WP-44K > PO Box 4 > West Point, PA 19486 > PH 215-652-2501 > fax. 215-993-6803 > e-mail. brett_connolly@merck.com > > > > ------------------------------------------------------------------------------ > Notice: This e-mail message, together with any attachments, contains > information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New > Jersey, USA 08889), and/or its affiliates (which may be known outside the > United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as > Banyu) that may be confidential, proprietary copyrighted and/or legally > privileged. It is intended solely for the use of the individual or entity > named on this message. If you are not the intended recipient, and have > received this message in error, please notify us immediately by reply > e-mail and then delete it from your system. > ------------------------------------------------------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ROrr <@t> enh.org Fri Jan 20 08:53:55 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Jan 20 08:54:06 2006 Subject: [Histonet] Cat Hematoxylin Message-ID: Angela, I use CAT hematoxylin as the counterstain on all of my IHC single and doublestains. It is a special formulation that does not need filtering. As I recall, the CAT denotes the initials of the scientists who formulated it, Cherukian, Allison, Tacha. I buy mine from Biocare Medical. www.biocare.net Let me know how your double stains are working. We use a few routinely, here. Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 From JPR <@t> Stowers-Institute.org Fri Jan 20 08:57:40 2006 From: JPR <@t> Stowers-Institute.org (Rey, Jean-Philippe) Date: Fri Jan 20 08:58:08 2006 Subject: [Histonet] Need Some Heat! Message-ID: Mike, Thermo is offering Shandon PARA trimmer, a "paraffin melter" that we are currently using in the lab to trim off the excess of paraffin from our block. It is really efficient and honestly less messy that the wax chips scattering everywhere. We usually use plastic cup to collect melting wax. Have a good one. JP REY -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mike Cupello Sent: Thursday, January 19, 2006 11:30 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need Some Heat! Hi to all of those out in Histoland... I was hoping to get some help. I have an employee who is suffering from tendonitis, and she feels that scraping blocks before trimming is adding to her injury. I know that there is some sort of device "out there" that uses heat to melt the excess wax from the outside of the block so that it will fit into the microtome chuck. Does anyone know of a type of heat block or hot knife that could accomplish this? I am looking for something that will collect the melted wax so that we don't have a huge mess to clean up after a day's work. Thanks for your time, Mike Cupello. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From froyer <@t> bitstream.net Fri Jan 20 09:07:10 2006 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Jan 20 09:07:30 2006 Subject: [Histonet] Need Some Heat! In-Reply-To: Message-ID: <003001c61dd3$307affb0$6f01a80a@fords> Ditto... Check out the Thermo Shandon "Para Trimmer", SKU (catalog number): B3120205 Web page: ~ Ford Ford M. Royer, MT(ASCP) Sales Manager, Histology Product Division Minnesota Medical Specialists, Inc. 7177 Madison Ave. W. Golden Valley, MN 55427-3601 763-542-8725 phone 888-790-9686 toll free 763-546-4830 fax email web -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rey, Jean-Philippe Sent: Friday, January 20, 2006 8:58 AM To: Mike Cupello; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Need Some Heat! Mike, Thermo is offering Shandon PARA trimmer, a "paraffin melter" that we are currently using in the lab to trim off the excess of paraffin from our block. It is really efficient and honestly less messy that the wax chips scattering everywhere. We usually use plastic cup to collect melting wax. Have a good one. JP REY -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mike Cupello Sent: Thursday, January 19, 2006 11:30 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need Some Heat! Hi to all of those out in Histoland... I was hoping to get some help. I have an employee who is suffering from tendonitis, and she feels that scraping blocks before trimming is adding to her injury. I know that there is some sort of device "out there" that uses heat to melt the excess wax from the outside of the block so that it will fit into the microtome chuck. Does anyone know of a type of heat block or hot knife that could accomplish this? I am looking for something that will collect the melted wax so that we don't have a huge mess to clean up after a day's work. Thanks for your time, Mike Cupello. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Jan 20 09:37:16 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Jan 20 09:37:29 2006 Subject: [Histonet] Re: IHC slides and sections In-Reply-To: <8C7EB4AA681C7C3-F0C-22C7@MBLK-M42.sysops.aol.com> References: <29B25753F6B1D51196110002A589D444032C49B1@PALMSG30.us.schp.com> <8C7EB4AA681C7C3-F0C-22C7@MBLK-M42.sysops.aol.com> Message-ID: <6.0.0.22.1.20060120083318.01b62688@gemini.msu.montana.edu> Erie's new EXEL slides (I may have spelled that incorrectly), they also make slides for microarray work - give them a call. Newcomers Supply has slides they maintain are more than just Plus charge - they have a website. Superfrost Gold although these are touted for use with frozen sections and not necessarily paraffin sections, but they are as described, pricey little gold nuggets. Some have used these for paraffin and thought it improved holding onto section. You can find these comments in Histonet Archives. At 01:04 PM 1/19/2006, you wrote: > > So what slides are the most adhesive? I only knew about the Superfrost + > slides. Are there more brands?? > >Lorie > >******************************************************************* >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. >******************************************************************* > > >-----Original Message----- >From: Osborn, Sharon >To: 'histonet@lists.utsouthwestern.edu' >Sent: Thu, 19 Jan 2006 13:36:54 -0500 >Subject: [Histonet] IHC slides and sections > > >Thanks a bunch for all the wonderful suggestions. We tried several of the >suggestions and sections still came off. The charged slides were still 'in >date'. We switched to a different slide and the sections remaned on. The >charged slides have different "stickiness" depending upon the vendor. In >the past we have used some slides that were 4-5 years old and the >stickiness was still present-excellent charge retention I guess. However, >we now have solved the problem with a stickier slide on the charge. Your >suggestions are helpful for our use overall and several are being implement >as a regular protocol. > >Have a great weekend! And, again THANKS! > >Sharon Osborn >DNAX, SP BioPharma >Palo Alto, CA >650-496-6539 > > >********************************************************************* >This message and any attachments are solely for the intended recipient. If >you >are not the intended recipient, disclosure, copying, use or distribution >of the >information included in this message is prohibited -- Please immediately and >permanently delete. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From gcallis <@t> montana.edu Fri Jan 20 09:48:55 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Jan 20 09:49:05 2006 Subject: [Histonet] Cat Hematoxylin In-Reply-To: <001f01c61d5b$b7672ee0$e8bd0b43@yourxhtr8hvc4p> References: <001f01c61d5b$b7672ee0$e8bd0b43@yourxhtr8hvc4p> Message-ID: <6.0.0.22.1.20060120084359.01b6b738@gemini.msu.montana.edu> It has something to do with a "Masterpiece" i.e. they (BIocare) also have something using Degas in the name. Trying to figure out who CAT is that produced masterpiece paintings, sculpture or whatever??? Want to bet it is some kind of progressive hematoxylin like Mayers, Gill recipes. Maybe they will step forward and explain the "artsy" nomenclature. At 05:51 PM 1/19/2006, you wrote: >I think Biocare Medical sells CAT hematoxylin. It's an abbreviation for >something, I forget. Then again, I've been forgetting a lot of things >lately. What were talking about? Oh, yeah, stop sniffing the xylene. Has >to be the xylene. > >Joe Nocito BS, HT(ASCP)QIHC >Histology Manager >Pathology Reference Lab >San Antonio, TX > >----- Original Message ----- From: "Angela Bitting" >To: >Sent: Thursday, January 19, 2006 2:38 PM >Subject: [Histonet] Cat Hematoxylin > > >I'm looking at a double-staining protocol that uses Cat Hematoxylin. Does >anyone know what Cat Hematoxylin is? I don't work in research, so maybe >this is something new or more widely utilized in a research setting. I >have a feeling that I'm going to feel really stupid when I hear the >answer, but I can't let this go. I need to know. Thanks, in advance, for >enlightening me. > > > >IMPORTANT WARNING: The information in this message (and the documents >attached to it, if any) is confidential and may be legally privileged. It >is intended solely for the addressee. Access to this message by anyone >else is unauthorized. If you are not the intended recipient, any >disclosure, copying, distribution or any action taken, or omitted to be >taken, in reliance on it is prohibited and may be unlawful. If you have >received this message in error, please delete all electronic copies of >this message (and the documents attached to it, if any), destroy any hard >copies you may have created and notify me immediately by replying to this >email. Thank you. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >-- >No virus found in this incoming message. >Checked by AVG Free Edition. >Version: 7.1.375 / Virus Database: 267.14.20/234 - Release Date: 1/18/2006 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Fri Jan 20 10:00:27 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Jan 20 10:00:40 2006 Subject: [Histonet] Need Some Heat! In-Reply-To: <001d01c61d82$994863d0$6401a8c0@HOME> References: <001d01c61d82$994863d0$6401a8c0@HOME> Message-ID: <6.0.0.22.1.20060120085111.01b69d18@gemini.msu.montana.edu> Mike, I have a young lady in the same boat, but her employer is doing nothing about it although the following device has been recommended. She resorted to embedding so the paraffin amount is just enough to not create this problem - a tedious way to do it, but it does work - however others embedding are not so careful. Go to Thermo Electron and look for their heated paraffin trimmer with slanted, heated surface and paraffin capture tray. (ParaTrimmer, not sure of the name) - it is a bit pricey, but well worth the money and I would perosnally love to have one in our lab. You employee's problem may stem from more than just scraping blocks, and you need to look at other ergonomics involved with her microtomy, embedding ,etc etc. Does she operate the microtome in an ergonomic manner, have an ergonomic chair, decent setup for reach, forceps that help her out, all those good things. Also what she does outside the lab may be a factor as the young lady here does some other physical activities (lots of cell phone dialing is one using her thumb to punch in the numbers) that contributes to her problems in the lab. Hopefully Jan Minshew, an expert on ergonomics in the histology laboratory can help you here too. At 10:30 PM 1/19/2006, you wrote: >Hi to all of those out in Histoland... > >I was hoping to get some help. I have an employee who is suffering from >tendonitis, and she feels that scraping blocks before trimming is adding >to her injury. I know that there is some sort of device "out there" that >uses heat to melt the excess wax from the outside of the block so that it >will fit into the microtome chuck. Does anyone know of a type of heat >block or hot knife that could accomplish this? I am looking for something >that will collect the melted wax so that we don't have a huge mess to >clean up after a day's work. > >Thanks for your time, > >Mike Cupello. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From vazquezr <@t> ohsu.edu Fri Jan 20 10:10:48 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Jan 20 10:11:21 2006 Subject: [Histonet] Need Some Heat! Message-ID: Mike, Why don't you appoint someone to scrap her blocks or all of them, until her tendonitis heals? We used to have a non-tech do this task and it would save us time. Tendonitis is only temporary generally. Robyn OHSU From jqb7 <@t> cdc.gov Fri Jan 20 10:13:57 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Jan 20 10:17:33 2006 Subject: [Histonet] Need Some Heat! Message-ID: The part number is B3120205 and it is called a Para Trimmer...they run around $500.00. But honestly, there is no need for scraping blocks if you embed carefully. I haven't had to scrape a block (except when I accidentally tipped one over on the embedding center cooling tray) in years. Jeanine Bartlett, HT(ASCP) (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 Gayle Callis Sent: Friday, January 20, 2006 11:00 AM To: Mike Cupello; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Need Some Heat! Mike, I have a young lady in the same boat, but her employer is doing nothing about it although the following device has been recommended. She resorted to embedding so the paraffin amount is just enough to not create this problem - a tedious way to do it, but it does work - however others embedding are not so careful. Go to Thermo Electron and look for their heated paraffin trimmer with slanted, heated surface and paraffin capture tray. (ParaTrimmer, not sure of the name) - it is a bit pricey, but well worth the money and I would perosnally love to have one in our lab. You employee's problem may stem from more than just scraping blocks, and you need to look at other ergonomics involved with her microtomy, embedding ,etc etc. Does she operate the microtome in an ergonomic manner, have an ergonomic chair, decent setup for reach, forceps that help her out, all those good things. Also what she does outside the lab may be a factor as the young lady here does some other physical activities (lots of cell phone dialing is one using her thumb to punch in the numbers) that contributes to her problems in the lab. Hopefully Jan Minshew, an expert on ergonomics in the histology laboratory can help you here too. At 10:30 PM 1/19/2006, you wrote: >Hi to all of those out in Histoland... > >I was hoping to get some help. I have an employee who is suffering >from tendonitis, and she feels that scraping blocks before trimming is >adding to her injury. I know that there is some sort of device "out >there" that uses heat to melt the excess wax from the outside of the >block so that it will fit into the microtome chuck. Does anyone know >of a type of heat block or hot knife that could accomplish this? I am >looking for something that will collect the melted wax so that we don't >have a huge mess to clean up after a day's work. > >Thanks for your time, > >Mike Cupello. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From adafeldman <@t> anatechltdusa.com Fri Jan 20 10:19:47 2006 From: adafeldman <@t> anatechltdusa.com (Ada Feldman) Date: Fri Jan 20 10:20:03 2006 Subject: [Histonet] Z fix In-Reply-To: References: Message-ID: <4FE5AD65-F73E-4DE6-BBD4-5043CD670FA1@anatechltdusa.com> Dear Betsy, Store the heart tissue in Z-FIX. Our in-house data show tissues stored in Z-FIX for 30 days show no deleterious effects on staining including immunohistochemistry. Sincerely, Ada T. Feldman, MS, HT/HTL(ASCP) Director of Manufacturing & Technical Services ANATECH LTD. 1020 Harts Lake Road Battle Creek, MI 49015 Phone: 800.262.8324 Fax: 269.964.8084 email: adafeldman@anatechltdusa.com website: www.anatechltdusa.com On Jan 19, 2006, at 1:49 PM, Molinari, Betsy wrote: > Hi, > > I have some mouse hearts in Z Fix. I am going to be out of town for > the > next week, can they stay in Z Fix or should I transfer them to 70%? > > Thanks. > > > > Betsy Molinari HT (ASCP) > > Texas Heart Institute > > Cardiovascular Pathology > > 6770 Bertner Ave. > > Houston,TX 77030 > > 832-355-6524 > > 832-355-6812 (fax) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dobbin <@t> upei.ca Fri Jan 20 12:04:30 2006 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Jan 20 12:12:48 2006 Subject: [Histonet] Anti-GRP-78 Message-ID: <43D0EDEF.7842.12104F7@acad1.cs.upei.ca> Hello Everyone, A colleague is about to stain (IHC) rat tissues (formalin fixed, paraffin embedded) for GRP-78. She has asked me to post her questions: Does anyone out there have previous experience with this marker? And if so, do you have any tips, especially with regard to antigen retrieval methods (if necessary). (She doing a lit search as I write!) Thanks so much. Greg From bjdewe <@t> aol.com Fri Jan 20 12:33:44 2006 From: bjdewe <@t> aol.com (bjdewe@aol.com) Date: Fri Jan 20 12:33:55 2006 Subject: [Histonet] Counter stain In-Reply-To: <6.0.0.22.1.20060120083007.01b608b8@gemini.msu.montana.edu> References: <8C7EB3D595FDB23-F0C-1832@MBLK-M42.sysops.aol.com> <6.0.0.22.1.20060120083007.01b608b8@gemini.msu.montana.edu> Message-ID: <8C7EC072D9D3EC9-14E0-502F@FWM-D34.sysops.aol.com> >>or not use any counterstain at all since the antigenic site is so small Yes, that is what I've done but they would be sooo pretty to photograph with a background stain... Cheers, Lorie ******************************************************************* 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. ******************************************************************* -----Original Message----- From: Gayle Callis To: bjdewe@aol.com Sent: Fri, 20 Jan 2006 08:30:59 -0700 Subject: Re: [Histonet] Counter stain Eosin? or not use any counterstain at all since the antigenic site is so small, hard to visualize after hematoxylin. At 11:28 AM 1/19/2006, you wrote: >I am doing a peroxidase DAB stain for an antigen that is present only in >small quantities in the nucleus. I am doing it on rat bone. If I >counterstain with hematoxylin it covers the DAB stain. What else can I use >that will only stain the cytoplasm and not the nucleus. i thought of >methyl green but it too is supposed to stain the nucleus. I thought of >neutral red but it won't look so good with the brown. Suggestions? > >Cheers, >Lorie > >******************************************************************* >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 Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From Jackie.O'Connor <@t> abbott.com Fri Jan 20 12:41:30 2006 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Jan 20 12:41:58 2006 Subject: [Histonet] Anti-GRP-78 Message-ID: Yep. I use a Santa Cruz goat poly at 1:400 with the Vector Elite HRP kit. I've tried it with both HIER and without - no difference - so no HIER is required. JO'C "Greg Dobbin" Sent by: histonet-bounces@lists.utsouthwestern.edu 01/20/2006 12:04 PM To: histonet@lists.utsouthwestern.edu cc: djones@avcn1.novell.upei.ca, (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Anti-GRP-78 Hello Everyone, A colleague is about to stain (IHC) rat tissues (formalin fixed, paraffin embedded) for GRP-78. She has asked me to post her questions: Does anyone out there have previous experience with this marker? And if so, do you have any tips, especially with regard to antigen retrieval methods (if necessary). (She doing a lit search as I write!) Thanks so much. Greg _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ROrr <@t> enh.org Fri Jan 20 13:38:46 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Jan 20 13:38:59 2006 Subject: [Histonet] Need some heat! Message-ID: It's the Thermo Shandon Para Trimmer. We have two. Ow they get hot! :) Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Friday, January 20, 2006 11:40 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 26, Issue 23 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: Cat Hematoxylin (Hofecker, Jennifer L) 2. Need Some Heat! (Mike Cupello) 3. marker of endothelial differentiation in vitro (Mark Adam Tarango) 4. please remove me, thanks! (mason dean) 5. Re: acetylcholinesterase staining (John Kiernan) 6. Re: Counter stain (John Kiernan) 7. Re: Invertebrates in Sediment (John Kiernan) 8. Re: air drying-white sections?? (John Kiernan) 9. Re: Need Some Heat! (gillian.2.brown@gsk.com) 10. job classification (Linda Blazek) 11. Lot to lot antibody validation (Connolly, Brett M) 12. slide storage feedback (Orr, Rebecca) 13. Re: Lot to lot antibody validation (Amy Porter) 14. Cat Hematoxylin (Orr, Rebecca) 15. RE: Need Some Heat! (Rey, Jean-Philippe) 16. RE: Need Some Heat! (Ford Royer) 17. Re: IHC slides and sections (Gayle Callis) 18. Re: Cat Hematoxylin (Gayle Callis) 19. Re: Need Some Heat! (Gayle Callis) 20. Re: Need Some Heat! (Robyn Vazquez) 21. RE: Need Some Heat! (Bartlett, Jeanine) 22. Re: Z fix (Ada Feldman) ---------------------------------------------------------------------- Message: 1 Date: Thu, 19 Jan 2006 21:36:54 -0600 From: "Hofecker, Jennifer L" Subject: RE: [Histonet] Cat Hematoxylin To: "histonet" Message-ID: <898D946569A27444B65667A49C074052852B5F@mailbe06.mc.vanderbilt.edu> Content-Type: text/plain; charset="iso-8859-1" I may be remembering this incorrectly, but isn't the CAT hematoxylin named after the two wonderful men who developed it? Chuck Churukian and David Tacha? Yes, sold by biocare... I don't remember what the difference is but I would imagine that somebody from Biocare Medical could help, or you could contact Charles or David... Joe, aren't you a little afraid of what might happen to us with out those fumes : ) Have a great weekend, Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe Nocito Sent: Thu 1/19/2006 6:51 PM To: Angela Bitting; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Cat Hematoxylin I think Biocare Medical sells CAT hematoxylin. It's an abbreviation for something, I forget. Then again, I've been forgetting a lot of things lately. What were talking about? Oh, yeah, stop sniffing the xylene. Has to be the xylene. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Angela Bitting" To: Sent: Thursday, January 19, 2006 2:38 PM Subject: [Histonet] Cat Hematoxylin I'm looking at a double-staining protocol that uses Cat Hematoxylin. Does anyone know what Cat Hematoxylin is? I don't work in research, so maybe this is something new or more widely utilized in a research setting. I have a feeling that I'm going to feel really stupid when I hear the answer, but I can't let this go. I need to know. Thanks, in advance, for enlightening me. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.1.375 / Virus Database: 267.14.20/234 - Release Date: 1/18/2006 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Thu, 19 Jan 2006 21:30:17 -0800 From: "Mike Cupello" Subject: [Histonet] Need Some Heat! To: Message-ID: <001d01c61d82$994863d0$6401a8c0@HOME> Content-Type: text/plain; charset="iso-8859-1" Hi to all of those out in Histoland... I was hoping to get some help. I have an employee who is suffering from tendonitis, and she feels that scraping blocks before trimming is adding to her injury. I know that there is some sort of device "out there" that uses heat to melt the excess wax from the outside of the block so that it will fit into the microtome chuck. Does anyone know of a type of heat block or hot knife that could accomplish this? I am looking for something that will collect the melted wax so that we don't have a huge mess to clean up after a day's work. Thanks for your time, Mike Cupello. ------------------------------ Message: 3 Date: Thu, 19 Jan 2006 20:45:14 -0800 (GMT-08:00) From: Mark Adam Tarango Subject: [Histonet] marker of endothelial differentiation in vitro To: histonet@lists.utsouthwestern.edu, DKAEK@coloplast.com Message-ID: <10756775.1137732314465.JavaMail.root@elwamui-karabash.atl.sa.earthlink. net> Content-Type: text/plain; charset=us-ascii okay, for endothelial cells in vitro try using CD31. I know nothing about eosin, but CD31 definately is specific for endothelial cells. Mark Tarango Date: Mon, 16 Jan 2006 11:05:28 +0100 From: Annette Ekblond Subject: [Histonet] marker of endothelial differentiation in vitro To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Dear subscribers -does anyone know of an endothelial marker specific to endothelial cells that have differentiated into tube-like structures (in vitro) - and not proliferating endothelial cultures?? Kind regards Annette CR Humlebaek Denmark ------------------------------ Message: 4 Date: Thu, 19 Jan 2006 23:40:19 -0800 From: mason dean Subject: [Histonet] please remove me, thanks! To: histonet@lists.utsouthwestern.edu Message-ID: <469494EB-9102-4DA6-A797-02496F9CAEA9@uci.edu> Content-Type: text/plain ------------------------------ Message: 5 Date: Fri, 20 Jan 2006 02:48:02 -0500 From: John Kiernan Subject: Re: [Histonet] acetylcholinesterase staining To: Angela Bitting Cc: histonet@lists.utsouthwestern.edu Message-ID: <43D095B2.75BDD11F@uwo.ca> Content-Type: text/plain; charset=us-ascii The dire warning following your enquiry is discouraging! Nevertheless, I make bold to reply. I have worked in the esterase histochemistry field, especially choline esterases, since the early 1960s, when most of the principles had been established. Which method is failing to work for you half of the time? These methods are extremely reliable if you go by the book (any book). Complete substrate solutions cannot be kept and used next day; that's a rule that applies to just about all enzyme activity histochemistry. John Kiernan Anatomy, UWO London, Canada ----------------------- Angela Bitting wrote: > > Our track record here with our acetylcholinesterase stain is about 50/50. We make up all the solutions fresh on Monday and it works fine, by Tuesday ...nothing. Can anyone send me a good reliable Acetylcholinesterase staining method? Maybe someone elses will work better for us. > > IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Fri, 20 Jan 2006 02:48:57 -0500 From: John Kiernan Subject: Re: [Histonet] Counter stain To: bjdewe@aol.com Cc: Histonet@lists.utsouthwestern.edu Message-ID: <43D095E9.BB2F937B@uwo.ca> Content-Type: text/plain; charset=us-ascii The best contrasting colour for DAB-brown is green. Ethyl green stains nuclei, so avoid it. (Methyl green has not been made for 25+ years. Dyes sold as "methyl green" should be ethyl green, which is better. Check with the vendor, because real methyl green won't perform properly without laborious purification by you in your lab.) For counterstaining cytoplasm but not nuclei, try fast green FCF. Instructions can be found in any textbook published since about 1940. Light green SF (pre-1940) acts similarly, but fades after a few years. John Kiernan Anatomy, UWO London, Canada ____________________________________________ bjdewe@aol.com wrote: > > I am doing a peroxidase DAB stain for an antigen that is present only in small quantities in the nucleus. I am doing it on rat bone. If I counterstain with hematoxylin it covers the DAB stain. What else can I use that will only stain the cytoplasm and not the nucleus. i thought of methyl green but it too is supposed to stain the nucleus. I thought of neutral red but it won't look so good with the brown. Suggestions? > > Cheers, > Lorie > > ******************************************************************* > 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 ------------------------------ Message: 7 Date: Fri, 20 Jan 2006 02:50:01 -0500 From: John Kiernan Subject: Re: [Histonet] Invertebrates in Sediment To: BennettW@pac.dfo-mpo.gc.ca Cc: Histonet Message-ID: <43D09629.3D8C3B2B@uwo.ca> Content-Type: text/plain; charset=us-ascii Rose Bengal B is an anionic, lipophilic xanthene dye, related to the eosins and erythrosins. One of its documented uses is staining foraminifera in marine sediments. Get your boss to send you to the library to check out "Conn's Biological Stains" and Clark's "Staining Procedures" for references to the original methods. If it's a specialized application, go to the references cited in these books. John Kiernan London, Canada. ______________________________________________________ BennettW@pac.dfo-mpo.gc.ca wrote: > > Hello Histonetters, > > Thanks in advance for spending any time with this query! I have a > researcher that has collected sediment samples, preserved with NBF, and > wants to stain any invertebrates in the sediment so that they can be > enumerated and identified. The researcher wants to use rose bengal but > doesn't know at what concentration and neither do I. Can anyone help? > > Thanks > Bill Bennett > Histologist > Fisheries and Oceans Canada > Pacific Biological Station > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Fri, 20 Jan 2006 02:51:16 -0500 From: John Kiernan Subject: Re: [Histonet] air drying-white sections?? To: Steven Coakley Cc: Histonet@lists.utsouthwestern.edu Message-ID: <43D09674.D41BE103@uwo.ca> Content-Type: text/plain; charset=us-ascii Dear Steven Coakley, Please explain how you "air dry my sections before drying them". Also, describe exactly what you do with the kimwipe (assuming that a kimwipe is a small paper hanky). If you're working with paraffin sections, it's important to let all the water drain off the slides (vertically) before letting the temperature rise to anywhere near the softening point of the wax. Histology waxes with 58C "melting points" soften at about 45C. If you use a hotplate, it gets uncomfortably hot for an applied hand in 10-20 seconds at 45C. That's the maximum allowable temperature before making a decision to melt the wax. The film of water between the ribbon and the glass must be all gone before you even think about melting the wax. This advice is in every textbook of histotechnology (microtechnique) published since about 1880, and it has appeared often in Histonet for ?10 years. The archives at http://histosearch.com are full of wisdom in this field. John Kiernan Anatomy, UWO London, Canada ____________________ Steven Coakley wrote: > > I've noticed that when I air dry my sections before drying them some of them are turning a chauky white. Might that indicate anything important. I really have not noticed this before in other places I work. I wipe off as much excess water with a kimwipe prior to letting the sections air dry vertically. Any thoughts. > > Steve =========== ------------------------------ Message: 9 Date: Fri, 20 Jan 2006 08:50:37 +0000 From: gillian.2.brown@gsk.com Subject: Re: [Histonet] Need Some Heat! To: "Mike Cupello" Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Hi Mike, we use a cheap domestic iron (set on delicates), standing in a shallow collecting dish. Gill Brown GlaxoSmithKline Medicines Research Centre, UK "Mike Cupello" Sent by: histonet-bounces@lists.utsouthwestern.edu 20-Jan-2006 05:30 To histonet@lists.utsouthwestern.edu cc Subject [Histonet] Need Some Heat! Hi to all of those out in Histoland... I was hoping to get some help. I have an employee who is suffering from tendonitis, and she feels that scraping blocks before trimming is adding to her injury. I know that there is some sort of device "out there" that uses heat to melt the excess wax from the outside of the block so that it will fit into the microtome chuck. Does anyone know of a type of heat block or hot knife that could accomplish this? I am looking for something that will collect the melted wax so that we don't have a huge mess to clean up after a day's work. Thanks for your time, Mike Cupello. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 10 Date: Fri, 20 Jan 2006 08:22:45 -0500 From: "Linda Blazek" Subject: [Histonet] job classification To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Does anyone have a dual classification of LIS manager and bench tech? If so is the salary higher than just a bench tech? Thanks Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org ------------------------------ Message: 11 Date: Fri, 20 Jan 2006 09:09:24 -0500 From: "Connolly, Brett M" Subject: [Histonet] Lot to lot antibody validation To: histonet@lists.utsouthwestern.edu Message-ID: <355C35514FEAC9458F75947F5270974D67CCA5@usctmx1103.merck.com> Content-Type: text/plain I would like to hear what procedures people are using to validate different lots of antibodies, including # of specimens you might use for the validation. Thanks, Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com ------------------------------------------------------------------------ ------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------ ------ ------------------------------ Message: 12 Date: Fri, 20 Jan 2006 08:43:52 -0600 From: "Orr, Rebecca" Subject: [Histonet] slide storage feedback To: Message-ID: Content-Type: text/plain; charset="us-ascii" Thanks everyone for your response! I got a lot of advice on long term slide storage sent to me privately, so just to share, I would say the general consensus was ultra low temps wrapped in foil along with a desiccant. Thanks again to the wonderful Mr. Tim Morken for sharing this article on the TMA storage and the paraffin "dip". The technique was what the Pathologist was looking for and what we will be using in our project. The Histonet is such a valuable tool, and I appreciate the support and help I have gotten from peers as well as vendors. Have the best weekend possible! I'll be spending my Saturday shoveling snow. Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 ------------------------------ Message: 21 Date: Thu, 19 Jan 2006 15:50:02 -0600 WS ------------------------------ Message: 13 Date: Fri, 20 Jan 2006 09:45:41 -0500 From: "Amy Porter" Subject: Re: [Histonet] Lot to lot antibody validation To: "Connolly, Brett M" , Message-ID: <000301c61dd0$2fd29210$8e7a0923@HistoJJ> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original We use a known postive control and run a system negative (no primary) with each new lot of primary that is received. We only run it one time maybe at the dilution for the old lot then slightly higher and slightly lower dilution with the already known pretreatment. Slides are then reviewed and the acceptable dilution for the new lot is recorded on an "Antibody Titration Log Sheet" signed off on by the pathologist or researcher to whom the work is being provided for. Hope this helps out... ----- Original Message ----- From: "Connolly, Brett M" To: Sent: Friday, January 20, 2006 9:09 AM Subject: [Histonet] Lot to lot antibody validation >I would like to hear what procedures people are using to validate different > lots of antibodies, including # of specimens you might use for the > validation. > > Thanks, Brett > > Brett M. Connolly, Ph.D. > Merck & Co., Inc. > MRL, Imaging Research > WP-44K > PO Box 4 > West Point, PA 19486 > PH 215-652-2501 > fax. 215-993-6803 > e-mail. brett_connolly@merck.com > > > > ------------------------------------------------------------------------ ------ > Notice: This e-mail message, together with any attachments, contains > information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New > Jersey, USA 08889), and/or its affiliates (which may be known outside the > United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as > Banyu) that may be confidential, proprietary copyrighted and/or legally > privileged. It is intended solely for the use of the individual or entity > named on this message. If you are not the intended recipient, and have > received this message in error, please notify us immediately by reply > e-mail and then delete it from your system. > ------------------------------------------------------------------------ ------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 14 Date: Fri, 20 Jan 2006 08:53:55 -0600 From: "Orr, Rebecca" Subject: [Histonet] Cat Hematoxylin To: Message-ID: Content-Type: text/plain; charset="us-ascii" Angela, I use CAT hematoxylin as the counterstain on all of my IHC single and doublestains. It is a special formulation that does not need filtering. As I recall, the CAT denotes the initials of the scientists who formulated it, Cherukian, Allison, Tacha. I buy mine from Biocare Medical. www.biocare.net Let me know how your double stains are working. We use a few routinely, here. Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 ------------------------------ Message: 15 Date: Fri, 20 Jan 2006 08:57:40 -0600 From: "Rey, Jean-Philippe" Subject: RE: [Histonet] Need Some Heat! To: "Mike Cupello" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Mike, Thermo is offering Shandon PARA trimmer, a "paraffin melter" that we are currently using in the lab to trim off the excess of paraffin from our block. It is really efficient and honestly less messy that the wax chips scattering everywhere. We usually use plastic cup to collect melting wax. Have a good one. JP REY -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mike Cupello Sent: Thursday, January 19, 2006 11:30 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need Some Heat! Hi to all of those out in Histoland... I was hoping to get some help. I have an employee who is suffering from tendonitis, and she feels that scraping blocks before trimming is adding to her injury. I know that there is some sort of device "out there" that uses heat to melt the excess wax from the outside of the block so that it will fit into the microtome chuck. Does anyone know of a type of heat block or hot knife that could accomplish this? I am looking for something that will collect the melted wax so that we don't have a huge mess to clean up after a day's work. Thanks for your time, Mike Cupello. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Fri, 20 Jan 2006 09:07:10 -0600 From: "Ford Royer" Subject: RE: [Histonet] Need Some Heat! To: Message-ID: <003001c61dd3$307affb0$6f01a80a@fords> Content-Type: text/plain; charset="us-ascii" Ditto... Check out the Thermo Shandon "Para Trimmer", SKU (catalog number): B3120205 Web page: ~ Ford Ford M. Royer, MT(ASCP) Sales Manager, Histology Product Division Minnesota Medical Specialists, Inc. 7177 Madison Ave. W. Golden Valley, MN 55427-3601 763-542-8725 phone 888-790-9686 toll free 763-546-4830 fax email web -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rey, Jean-Philippe Sent: Friday, January 20, 2006 8:58 AM To: Mike Cupello; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Need Some Heat! Mike, Thermo is offering Shandon PARA trimmer, a "paraffin melter" that we are currently using in the lab to trim off the excess of paraffin from our block. It is really efficient and honestly less messy that the wax chips scattering everywhere. We usually use plastic cup to collect melting wax. Have a good one. JP REY -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mike Cupello Sent: Thursday, January 19, 2006 11:30 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need Some Heat! Hi to all of those out in Histoland... I was hoping to get some help. I have an employee who is suffering from tendonitis, and she feels that scraping blocks before trimming is adding to her injury. I know that there is some sort of device "out there" that uses heat to melt the excess wax from the outside of the block so that it will fit into the microtome chuck. Does anyone know of a type of heat block or hot knife that could accomplish this? I am looking for something that will collect the melted wax so that we don't have a huge mess to clean up after a day's work. Thanks for your time, Mike Cupello. _______________________________________________ Histonet mailing list Histonet@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: Fri, 20 Jan 2006 08:37:16 -0700 From: Gayle Callis Subject: [Histonet] Re: IHC slides and sections To: bjdewe@aol.com, Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20060120083318.01b62688@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Erie's new EXEL slides (I may have spelled that incorrectly), they also make slides for microarray work - give them a call. Newcomers Supply has slides they maintain are more than just Plus charge - they have a website. Superfrost Gold although these are touted for use with frozen sections and not necessarily paraffin sections, but they are as described, pricey little gold nuggets. Some have used these for paraffin and thought it improved holding onto section. You can find these comments in Histonet Archives. At 01:04 PM 1/19/2006, you wrote: > > So what slides are the most adhesive? I only knew about the Superfrost + > slides. Are there more brands?? > >Lorie > >******************************************************************* >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. >******************************************************************* > > >-----Original Message----- >From: Osborn, Sharon >To: 'histonet@lists.utsouthwestern.edu' >Sent: Thu, 19 Jan 2006 13:36:54 -0500 >Subject: [Histonet] IHC slides and sections > > >Thanks a bunch for all the wonderful suggestions. We tried several of the >suggestions and sections still came off. The charged slides were still 'in >date'. We switched to a different slide and the sections remaned on. The >charged slides have different "stickiness" depending upon the vendor. In >the past we have used some slides that were 4-5 years old and the >stickiness was still present-excellent charge retention I guess. However, >we now have solved the problem with a stickier slide on the charge. Your >suggestions are helpful for our use overall and several are being implement >as a regular protocol. > >Have a great weekend! And, again THANKS! > >Sharon Osborn >DNAX, SP BioPharma >Palo Alto, CA >650-496-6539 > > >********************************************************************* >This message and any attachments are solely for the intended recipient. If >you >are not the intended recipient, disclosure, copying, use or distribution >of the >information included in this message is prohibited -- Please immediately and >permanently delete. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 ------------------------------ Message: 18 Date: Fri, 20 Jan 2006 08:48:55 -0700 From: Gayle Callis Subject: Re: [Histonet] Cat Hematoxylin To: "Joe Nocito" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20060120084359.01b6b738@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed It has something to do with a "Masterpiece" i.e. they (BIocare) also have something using Degas in the name. Trying to figure out who CAT is that produced masterpiece paintings, sculpture or whatever??? Want to bet it is some kind of progressive hematoxylin like Mayers, Gill recipes. Maybe they will step forward and explain the "artsy" nomenclature. At 05:51 PM 1/19/2006, you wrote: >I think Biocare Medical sells CAT hematoxylin. It's an abbreviation for >something, I forget. Then again, I've been forgetting a lot of things >lately. What were talking about? Oh, yeah, stop sniffing the xylene. Has >to be the xylene. > >Joe Nocito BS, HT(ASCP)QIHC >Histology Manager >Pathology Reference Lab >San Antonio, TX > >----- Original Message ----- From: "Angela Bitting" >To: >Sent: Thursday, January 19, 2006 2:38 PM >Subject: [Histonet] Cat Hematoxylin > > >I'm looking at a double-staining protocol that uses Cat Hematoxylin. Does >anyone know what Cat Hematoxylin is? I don't work in research, so maybe >this is something new or more widely utilized in a research setting. I >have a feeling that I'm going to feel really stupid when I hear the >answer, but I can't let this go. I need to know. Thanks, in advance, for >enlightening me. > > > >IMPORTANT WARNING: The information in this message (and the documents >attached to it, if any) is confidential and may be legally privileged. It >is intended solely for the addressee. Access to this message by anyone >else is unauthorized. If you are not the intended recipient, any >disclosure, copying, distribution or any action taken, or omitted to be >taken, in reliance on it is prohibited and may be unlawful. If you have >received this message in error, please delete all electronic copies of >this message (and the documents attached to it, if any), destroy any hard >copies you may have created and notify me immediately by replying to this >email. Thank you. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >-- >No virus found in this incoming message. >Checked by AVG Free Edition. >Version: 7.1.375 / Virus Database: 267.14.20/234 - Release Date: 1/18/2006 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 19 Date: Fri, 20 Jan 2006 09:00:27 -0700 From: Gayle Callis Subject: Re: [Histonet] Need Some Heat! To: "Mike Cupello" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20060120085111.01b69d18@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Mike, I have a young lady in the same boat, but her employer is doing nothing about it although the following device has been recommended. She resorted to embedding so the paraffin amount is just enough to not create this problem - a tedious way to do it, but it does work - however others embedding are not so careful. Go to Thermo Electron and look for their heated paraffin trimmer with slanted, heated surface and paraffin capture tray. (ParaTrimmer, not sure of the name) - it is a bit pricey, but well worth the money and I would perosnally love to have one in our lab. You employee's problem may stem from more than just scraping blocks, and you need to look at other ergonomics involved with her microtomy, embedding ,etc etc. Does she operate the microtome in an ergonomic manner, have an ergonomic chair, decent setup for reach, forceps that help her out, all those good things. Also what she does outside the lab may be a factor as the young lady here does some other physical activities (lots of cell phone dialing is one using her thumb to punch in the numbers) that contributes to her problems in the lab. Hopefully Jan Minshew, an expert on ergonomics in the histology laboratory can help you here too. At 10:30 PM 1/19/2006, you wrote: >Hi to all of those out in Histoland... > >I was hoping to get some help. I have an employee who is suffering from >tendonitis, and she feels that scraping blocks before trimming is adding >to her injury. I know that there is some sort of device "out there" that >uses heat to melt the excess wax from the outside of the block so that it >will fit into the microtome chuck. Does anyone know of a type of heat >block or hot knife that could accomplish this? I am looking for something >that will collect the melted wax so that we don't have a huge mess to >clean up after a day's work. > >Thanks for your time, > >Mike Cupello. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 20 Date: Fri, 20 Jan 2006 08:10:48 -0800 From: "Robyn Vazquez" Subject: Re: [Histonet] Need Some Heat! To: histonet@lists.utsouthwestern.edu, mike_cup@telus.net Message-ID: Content-Type: text/plain; charset=us-ascii Mike, Why don't you appoint someone to scrap her blocks or all of them, until her tendonitis heals? We used to have a non-tech do this task and it would save us time. Tendonitis is only temporary generally. Robyn OHSU ------------------------------ Message: 21 Date: Fri, 20 Jan 2006 11:13:57 -0500 From: "Bartlett, Jeanine" Subject: RE: [Histonet] Need Some Heat! To: "Gayle Callis" , "Mike Cupello" , Message-ID: Content-Type: text/plain; charset="US-ASCII" The part number is B3120205 and it is called a Para Trimmer...they run around $500.00. But honestly, there is no need for scraping blocks if you embed carefully. I haven't had to scrape a block (except when I accidentally tipped one over on the embedding center cooling tray) in years. Jeanine Bartlett, HT(ASCP) (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 Gayle Callis Sent: Friday, January 20, 2006 11:00 AM To: Mike Cupello; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Need Some Heat! Mike, I have a young lady in the same boat, but her employer is doing nothing about it although the following device has been recommended. She resorted to embedding so the paraffin amount is just enough to not create this problem - a tedious way to do it, but it does work - however others embedding are not so careful. Go to Thermo Electron and look for their heated paraffin trimmer with slanted, heated surface and paraffin capture tray. (ParaTrimmer, not sure of the name) - it is a bit pricey, but well worth the money and I would perosnally love to have one in our lab. You employee's problem may stem from more than just scraping blocks, and you need to look at other ergonomics involved with her microtomy, embedding ,etc etc. Does she operate the microtome in an ergonomic manner, have an ergonomic chair, decent setup for reach, forceps that help her out, all those good things. Also what she does outside the lab may be a factor as the young lady here does some other physical activities (lots of cell phone dialing is one using her thumb to punch in the numbers) that contributes to her problems in the lab. Hopefully Jan Minshew, an expert on ergonomics in the histology laboratory can help you here too. At 10:30 PM 1/19/2006, you wrote: >Hi to all of those out in Histoland... > >I was hoping to get some help. I have an employee who is suffering >from tendonitis, and she feels that scraping blocks before trimming is >adding to her injury. I know that there is some sort of device "out >there" that uses heat to melt the excess wax from the outside of the >block so that it will fit into the microtome chuck. Does anyone know >of a type of heat block or hot knife that could accomplish this? I am >looking for something that will collect the melted wax so that we don't >have a huge mess to clean up after a day's work. > >Thanks for your time, > >Mike Cupello. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 22 Date: Fri, 20 Jan 2006 11:19:47 -0500 From: Ada Feldman Subject: Re: [Histonet] Z fix To: "Molinari, Betsy" Cc: histonet@lists.utsouthwestern.edu Message-ID: <4FE5AD65-F73E-4DE6-BBD4-5043CD670FA1@anatechltdusa.com> Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed Dear Betsy, Store the heart tissue in Z-FIX. Our in-house data show tissues stored in Z-FIX for 30 days show no deleterious effects on staining including immunohistochemistry. Sincerely, Ada T. Feldman, MS, HT/HTL(ASCP) Director of Manufacturing & Technical Services ANATECH LTD. 1020 Harts Lake Road Battle Creek, MI 49015 Phone: 800.262.8324 Fax: 269.964.8084 email: adafeldman@anatechltdusa.com website: www.anatechltdusa.com On Jan 19, 2006, at 1:49 PM, Molinari, Betsy wrote: > Hi, > > I have some mouse hearts in Z Fix. I am going to be out of town for > the > next week, can they stay in Z Fix or should I transfer them to 70%? > > Thanks. > > > > Betsy Molinari HT (ASCP) > > Texas Heart Institute > > Cardiovascular Pathology > > 6770 Bertner Ave. > > Houston,TX 77030 > > 832-355-6524 > > 832-355-6812 (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 End of Histonet Digest, Vol 26, Issue 23 **************************************** From PMonfils <@t> Lifespan.org Fri Jan 20 14:52:08 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Jan 20 14:52:20 2006 Subject: [Histonet] Need Some Heat! Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717651@lsexch.lsmaster.lifespan.org> Many years ago Technicon made an "electric paraffin knife", which actually looked more like a little hatchet than a knife. I found one in a box of "junk" that someone was throwing out about fifteen years ago, had our electrical department rewire it, and I've been using it ever since. It's a very valuable laboratory tool, and I always wondered why no other company started making such a device once Technicon stopped. Sounds like the Para Trimmer is the answer to that question. I frequently embed large paraffin blocks, using a variety of little boxes, plastic jars, etc. as embedding molds, and the paraffin knife trims and shapes such blocks very efficiently. From BennettW <@t> pac.dfo-mpo.gc.ca Fri Jan 20 15:34:11 2006 From: BennettW <@t> pac.dfo-mpo.gc.ca (BennettW@pac.dfo-mpo.gc.ca) Date: Fri Jan 20 15:31:01 2006 Subject: [Histonet] DNA PCR Question Message-ID: <7CBBD627E4E688499349A5D11D078316031309EE@msgpacpbs.pac.dfo-mpo.ca> Many thanks to the histonetters that answered my question about inverts in mud but I have another question in a field that I know nothing about. Why is it when people see paraffin they come running to the histologist? O.K. that's not the question. Here it is. I have a researcher (a different one from last time) that is trying to extract shellfish pathogen DNA from a paraffin embedded sample. He is getting background fluorescence in a RTPCR using DNA extracted from this paraffin embedded tissue. The tissue was originally fixed in Davidsons fixative. He is using a Takara Dexpat kit and he wants to know how to solve this problem using a Taqman MGB (FAM) probe? Any suggestions would be great and much appreciated. Cheers Bill Bennett Histologist Fisheries and Oceans Canada Pacific Biological Station From sjchtascp <@t> yahoo.com Fri Jan 20 15:40:17 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Jan 20 15:40:28 2006 Subject: [Histonet] EMS Maldonado's Message-ID: <20060120214017.58985.qmail@web90205.mail.scd.yahoo.com> Has anyone out there used this EMS kit with satisfactory results on pancreas tissue. Thanks, Steve --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. From pruegg <@t> ihctech.net Fri Jan 20 16:12:03 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Jan 20 16:12:09 2006 Subject: [Histonet] CSF1 Message-ID: <200601202211.k0KMBwVK004669@chip.viawest.net> I need to hear of any experiences with Colony Stimulating Factor (CSF1/CFems) antibody for IHC on FFPE tissues, please. I see that there are lots listed but I do not know which or if they work in FFPE tissues. Thank you, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From pruegg <@t> ihctech.net Fri Jan 20 17:11:32 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Jan 20 17:11:42 2006 Subject: [Histonet] air drying-white sections?? In-Reply-To: <43D09674.D41BE103@uwo.ca> Message-ID: <200601202311.k0KNBSVK021072@chip.viawest.net> Besides carefully standing vertically and tapping my slides dry after picking them up off the water bath, I put them in a rack on their sides and place them over a small fan box that my husband made for me. The fan is incased in a wire box and is about 4 inches square, the rack of 25 slides sits on top of the fan (this fan is very gentle, he put a reastat or something on it to reduce the motor power so it won't blow too hard) and let them dry/drain before I do any heating to melt the paraffin. I usually leave them on the fan for 10-20 min. then lay them flat on a heat plate set at 55dc for 30 min., I never have sections fall off unless they are cartilage and bone but that is a different issue altogether. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Friday, January 20, 2006 12:51 AM To: Steven Coakley Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] air drying-white sections?? Dear Steven Coakley, Please explain how you "air dry my sections before drying them". Also, describe exactly what you do with the kimwipe (assuming that a kimwipe is a small paper hanky). If you're working with paraffin sections, it's important to let all the water drain off the slides (vertically) before letting the temperature rise to anywhere near the softening point of the wax. Histology waxes with 58C "melting points" soften at about 45C. If you use a hotplate, it gets uncomfortably hot for an applied hand in 10-20 seconds at 45C. That's the maximum allowable temperature before making a decision to melt the wax. The film of water between the ribbon and the glass must be all gone before you even think about melting the wax. This advice is in every textbook of histotechnology (microtechnique) published since about 1880, and it has appeared often in Histonet for ?10 years. The archives at http://histosearch.com are full of wisdom in this field. John Kiernan Anatomy, UWO London, Canada ____________________ Steven Coakley wrote: > > I've noticed that when I air dry my sections before drying them some of them are turning a chauky white. Might that indicate anything important. I really have not noticed this before in other places I work. I wipe off as much excess water with a kimwipe prior to letting the sections air dry vertically. Any thoughts. > > Steve =========== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BMolinari <@t> heart.thi.tmc.edu Sat Jan 21 05:55:45 2006 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Sat Jan 21 06:01:28 2006 Subject: [Histonet] unsubscribe Message-ID: Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) From pruegg <@t> ihctech.net Sat Jan 21 11:53:12 2006 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Sat Jan 21 11:53:38 2006 Subject: [Histonet] anti goat labeled polymer Message-ID: <200601211753.k0LHrKeY000153@pro12.abac.com> Hey Guys, Has anyone found a good anti goat labeled polymer yet? I tried one a while ago but was not happy with it. Come on you vendors, we need this. I am even using goat antibodies for clinical work now so you can't use the excuse that only researchers use goat antibodies anymore. Thanks, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net website www.ihctech.net From anh2006 <@t> med.cornell.edu Sat Jan 21 12:15:38 2006 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Sat Jan 21 12:15:48 2006 Subject: [Histonet] anti goat labeled polymer In-Reply-To: <200601211753.k0LHrKeY000153@pro12.abac.com> References: <200601211753.k0LHrKeY000153@pro12.abac.com> Message-ID: A reliable anti-rat polymer would also be MUCH APPRECIATED. At 10:53 AM -0700 1/21/06, pruegg@ihctech.net wrote: >Hey Guys, > >Has anyone found a good anti goat labeled polymer yet? I tried one a while >ago but was not happy with it. Come on you vendors, we need this. I am even >using goat antibodies for clinical work now so you can't use the excuse that >only researchers use goat antibodies anymore. > >Thanks, > >Patsy > > > -- From marktarango <@t> earthlink.net Sat Jan 21 16:06:41 2006 From: marktarango <@t> earthlink.net (Mark Adam Tarango) Date: Sat Jan 21 16:06:49 2006 Subject: [Histonet] Re: marker of endothelial differentiation in vitro Message-ID: <27592686.1137881201836.JavaMail.root@elwamui-polski.atl.sa.earthlink.net> You know it's funny that you mention tube like structures...... do they look fairly long and rectangular? I have something growing in tissue culture that looks like that. These cells started off as hemangiomas. We used collangenase to break it up, and are growing these cells in DMEM with added penicillin/streptomyacin and fetal bovine serum. I've seen quite a few of these, one person thinks they look like myocytes....heart muscle cells in particular, they aren't beating or anythying though. And if that is what they are, i'd be really really surprised. Most of the cells look like fibroblasts (I'm thinking that what look like fibroblasts might be pericytes) with some scattered possible endothelial cells. We transfected a few dishes with a vector to induce division, but we don't see these cells in any of the tissue culture dishes containing the transformed cells. I wonder if we are looking at the same thing. I've ordered a camera for our inverted scope, i'll have to take a picture when it comes in. I'm curious if your "tube like structures" resemble what i have in culture. Can you send a picture? Do you see multple cells composing the tube-like structures? Are you sure that they are even cells? Sounds intersting. Mark Tarango Date: Mon, 16 Jan 2006 11:05:28 +0100 From: Annette Ekblond Subject: [Histonet] marker of endothelial differentiation in vitro To: histonet@lists.utsouthwestern.edu Dear subscribers -does anyone know of an endothelial marker specific to endothelial cells that have differentiated into tube-like structures (in vitro) - and not proliferating endothelial cultures?? Kind regards Annette CR Humlebaek Denmark From stevenk <@t> med.usyd.edu.au Sun Jan 22 22:58:51 2006 From: stevenk <@t> med.usyd.edu.au (stevenk) Date: Sun Jan 22 22:54:05 2006 Subject: [Histonet] Re: DNA Extraction from clotted blood Message-ID: I was wondering if anyone could help me with a reliable DNA Extraction method for clotted blood. Was interested in obtaining a good yield as well as quality DNA. Thanks Stephen -- Stephen Kum Jew Senior Technical Officer Discipline of Pathology School of Medical Sciences Blackburn Building D06 University of Sydney NSW 2006 Australia Ph: + 61 2 9351 6143 Fax:+61 2 9351 3429 From pndrew_arior <@t> yahoo.co.uk Mon Jan 23 02:47:21 2006 From: pndrew_arior <@t> yahoo.co.uk (Andrew Prior) Date: Mon Jan 23 02:47:30 2006 Subject: [Histonet] Bone/ metal sections Message-ID: <20060123084721.66740.qmail@web25708.mail.ukl.yahoo.com> Does anyone out there do thin (<40micron) sections of MMA embedded bone containing metal implants? If so, what?s your average turn-around time for say 2 sections from 4 blocks, not including embedding time? We are having our annual review and need to show that our productivity is in line with other labs. Thanks in advance Andrew Prior Histologist ___________________________________________________________ Yahoo! Messenger - NEW crystal clear PC to PC calling worldwide with voicemail http://uk.messenger.yahoo.com From eva-moeller <@t> arcor.de Mon Jan 23 05:55:04 2006 From: eva-moeller <@t> arcor.de (eva-moeller@arcor.de) Date: Mon Jan 23 05:55:17 2006 Subject: [Histonet] BrdU staining Message-ID: <7984516.1138017304028.JavaMail.ngmail@webmail-06.arcor-online.net> Hello everybody, I am trying to do a BrdU staining on frozen tissue slices (already fixed with PFA 4%) for weeks. The signal is restricted to the rim of the core and not within it. I am staining with anti-BrdU by the Roche detection kit. I already tried their protocol but that did not work on my fixed slices. I dry the slices then rehydrate for 1 h. After that I apply the 1st antibody for 30 min 37?C. Thanks for support. Eva Machen Sie aus 14 Cent spielend bis zu 100 Euro! Die neue Gaming-Area von Arcor - ?ber 50 Onlinespiele im Angebot. http://www.arcor.de/rd/emf-gaming-1 From mgdelaware <@t> comcast.net Mon Jan 23 08:21:13 2006 From: mgdelaware <@t> comcast.net (Marian powers) Date: Mon Jan 23 08:21:26 2006 Subject: [Histonet] Microwave processsing Message-ID: <000e01c62028$44408ea0$be4dff45@MEGAN> Hello, I am looking into microwave processors, could anyone recommend any? Thanks for your help. Sincerely, Marian Powers, HT(ASCP) Doctors Pathology Services 882 Walker Rd. Dover, DE 19904 (302) 677-0000 Fax (302) 677-0010 From rjbuesa <@t> yahoo.com Mon Jan 23 08:43:06 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Jan 23 08:43:15 2006 Subject: [Histonet] Microwave processsing In-Reply-To: <000e01c62028$44408ea0$be4dff45@MEGAN> Message-ID: <20060123144306.23071.qmail@web61218.mail.yahoo.com> Marian: You could try to contact "Microwave Research and Applications, Inc." This company manufacture small microwaves that have been commissioned and successfully used by several laboratories. You could contact the President, Mr. Wayne Love wloves@att.net or visit their web site www.microwaveresearch.com Hope this will help you! Ren? J. Marian powers wrote: Hello, I am looking into microwave processors, could anyone recommend any? Thanks for your help. Sincerely, Marian Powers, HT(ASCP) Doctors Pathology Services 882 Walker Rd. Dover, DE 19904 (302) 677-0000 Fax (302) 677-0010 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos ? Showcase holiday pictures in hardcover Photo Books. You design it and we?ll bind it! From ROrr <@t> enh.org Mon Jan 23 08:43:18 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Mon Jan 23 08:43:27 2006 Subject: [Histonet] goat polymers Message-ID: I'm with Patsy! Bring on the Goat polymer! Goat polyclonals are emerging! Go Steelers! Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 From DOOLEEO <@t> shands.ufl.edu Mon Jan 23 08:49:13 2006 From: DOOLEEO <@t> shands.ufl.edu (Elaine Dooley) Date: Mon Jan 23 08:48:48 2006 Subject: [Histonet] egfr variant 3 Message-ID: Dear Histonetters, Is there a source for this antibody through a research lab or commercial vendor? The company I was getting it from has discontinued it. Thanks in advance Elaine Dooley From pmcardle <@t> ebsciences.com Mon Jan 23 09:08:55 2006 From: pmcardle <@t> ebsciences.com (Phil McArdle) Date: Mon Jan 23 09:09:08 2006 Subject: [Histonet] Re: Microwave processing Message-ID: <43D4F187.3040101@ebsciences.com> Hi: Hope you don't mind a "vendor" e-mail, but you might want to check out www.ebsciences.com. Best regards, Phil McArdle -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" From Wanda.Smith <@t> HCAhealthcare.com Mon Jan 23 09:46:31 2006 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Mon Jan 23 09:46:40 2006 Subject: [Histonet] CAP Question ANP.27720 Microwave Leakage Message-ID: Good Morning Histonetters, I have a question about a question on the CAP Checklist. We are doing our Self-Evaluation and one of the new questions ANP.27720 "Are microwave devices monitored at least annually to ensure that there is less than 5mW/cm2 leakage at a distance of 5 cm from the surface?" Question: Who takes the measurement??? My Biomed Departmet does not and neither does my Engineering Department!!! I'm not sure where to go from here, so I thought.....THE HISTONET!!!!!!!!! Please Help! Wanda Wanda G. Smith, BHS, HTL(ASCP)HT Pathology Supervisor Trident Medical Center 9330 Medical Plaza Drive Charleston, SC 29406 843-797-4586 843-797-4296 fax wanda.smith@hcahealthcare.com From plucas <@t> biopath.org Mon Jan 23 09:58:52 2006 From: plucas <@t> biopath.org (Paula Lucas) Date: Mon Jan 23 09:57:59 2006 Subject: [Histonet] Job Opportunity - Fountain Valley California Message-ID: <006001c62035$e8ea69f0$7c01a8c0@biopath.org> Seeking a P/T Histotech to embed and cut tissue specimens. 5 am to 10 am We are a small private laboratory who services 2 hospitals and surgery centers in Fountain Valley, California (Orange County area). Paula Lucas Bio-Path Medical Group 17150 Newhope Street, #117 Fountain Valley, CA 92708 From rjbuesa <@t> yahoo.com Mon Jan 23 10:10:01 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Jan 23 10:10:12 2006 Subject: [Histonet] CAP Question ANP.27720 Microwave Leakage In-Reply-To: Message-ID: <20060123161001.45530.qmail@web61216.mail.yahoo.com> Wanda: At my hospital the Safety Officer was the one doing that task, and she did it annually in a very thorough way. She even tested MW ovens used in the lunch room. Ren? J. Smith Wanda wrote: Good Morning Histonetters, I have a question about a question on the CAP Checklist. We are doing our Self-Evaluation and one of the new questions ANP.27720 "Are microwave devices monitored at least annually to ensure that there is less than 5mW/cm2 leakage at a distance of 5 cm from the surface?" Question: Who takes the measurement??? My Biomed Departmet does not and neither does my Engineering Department!!! I'm not sure where to go from here, so I thought.....THE HISTONET!!!!!!!!! Please Help! Wanda Wanda G. Smith, BHS, HTL(ASCP)HT Pathology Supervisor Trident Medical Center 9330 Medical Plaza Drive Charleston, SC 29406 843-797-4586 843-797-4296 fax wanda.smith@hcahealthcare.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. From pmcardle <@t> ebsciences.com Mon Jan 23 10:24:59 2006 From: pmcardle <@t> ebsciences.com (Phil McArdle) Date: Mon Jan 23 10:25:18 2006 Subject: [Histonet] CAP Question ANP.27720 Microwave Leakage Message-ID: <43D5035B.4040808@ebsciences.com> Hi: Hope you don't mind a "vendor" e-mail, but you might want to check out the following link: http://www.ebsciences.com/pdf/EBS_CAP_RECOMMEND.pdf Best regards, Phil McArdle -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" From froyer <@t> bitstream.net Mon Jan 23 10:48:40 2006 From: froyer <@t> bitstream.net (Ford Royer) Date: Mon Jan 23 10:49:08 2006 Subject: [Histonet] Melan-A Staining Message-ID: <007701c6203c$dcfb2150$6f01a80a@fords> I consult for a small lab in my area and they are contemplating doing Melan-A staining in their lab. They are wondering if this would be feasible and what might the costs be. Could any of you out there in HistoLand be of assistance? They are estimating that they will do an average of one patient slide per day. How much do the reagents cost (cost per test)? Which manufacturers carry it? How many steps are involved in the procedure? How much total time is involved to perform the procedure? Does it take a high level of skill to perform? (They will be staining by hand for the time being) Etc.? This lab does not do any special staining and the present time. This will be a first for them, so any helpful hints will be greatly appreciated. As you can see, I am totally ignorant in this department, so I am of no help at all. Cheers! ~ Ford Ford M. Royer, MT(ASCP) Sales Manager, Histology Product Division Minnesota Medical Specialists, Inc. 7177 Madison Ave. W. Golden Valley, MN 55427-3601 763-542-8725 phone 888-790-9686 toll free 763-546-4830 fax email web From portera <@t> msu.edu Mon Jan 23 11:01:52 2006 From: portera <@t> msu.edu (Amy Porter) Date: Mon Jan 23 11:02:27 2006 Subject: [Histonet] CAP Question ANP.27720 Microwave Leakage References: Message-ID: <001b01c6203e$b543abd0$8e7a0923@HistoJJ> Maybe you could try someone who works in the radiology / or medical scanning area???? Just a thought! ----- Original Message ----- From: "Smith Wanda" To: "Histonet (E-mail)" Sent: Monday, January 23, 2006 10:46 AM Subject: [Histonet] CAP Question ANP.27720 Microwave Leakage Good Morning Histonetters, I have a question about a question on the CAP Checklist. We are doing our Self-Evaluation and one of the new questions ANP.27720 "Are microwave devices monitored at least annually to ensure that there is less than 5mW/cm2 leakage at a distance of 5 cm from the surface?" Question: Who takes the measurement??? My Biomed Departmet does not and neither does my Engineering Department!!! I'm not sure where to go from here, so I thought.....THE HISTONET!!!!!!!!! Please Help! Wanda Wanda G. Smith, BHS, HTL(ASCP)HT Pathology Supervisor Trident Medical Center 9330 Medical Plaza Drive Charleston, SC 29406 843-797-4586 843-797-4296 fax wanda.smith@hcahealthcare.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon Jan 23 11:50:37 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Jan 23 11:50:45 2006 Subject: [Histonet] Melan-A Staining In-Reply-To: <007701c6203c$dcfb2150$6f01a80a@fords> Message-ID: <20060123175037.82001.qmail@web61216.mail.yahoo.com> Hi Royer: I used Melan A Clone A103 (Ig Mouse) bought from DAKO at a cost (in 2002) of $319 per mL of concentrated antibody (their cat. no. M7196 then). I used it a a dilution of 1:50 after HIER at pH6 (with melanoma case as + control). This means that you could dilute it (not at once, but during all the shelf life as needed) to up to 50 mL of diluted Ab. By hand you can expect to use about 250 ?L/slide which means that you could treat about 200 slides = $1.60 per slide in antibody costs. You will need all the other reagents for a "conventional" protocol. Some years ago I calculated that materials per slide for an IHC procedure amounted to about $5.80 Labor, depending on the number os slides and practice, could be as low as $2.15 All in all you could expect to spend about $ 8 to $9/slide. In order to reduce costs, they could batch slides and do the procedure twice weeknly. You just need a good protocol and somebody with manual dexterity (as any histotech should have, by the way). Hope this will help, a questoin though, what do you mean that you "consult for a small lab in your area"? Ren? J. I consult for a small lab in my area and they are contemplating doing Melan-A staining in their lab. They are wondering if this would be feasible and what might the costs be. Could any of you out there in HistoLand be of assistance? They are estimating that they will do an average of one patient slide per day. How much do the reagents cost (cost per test)? Which manufacturers carry it? How many steps are involved in the procedure? How much total time is involved to perform the procedure? Does it take a high level of skill to perform? (They will be staining by hand for the time being) Etc.? This lab does not do any special staining and the present time. This will be a first for them, so any helpful hints will be greatly appreciated. As you can see, I am totally ignorant in this department, so I am of no help at all. Cheers! ~ Ford Ford M. Royer, MT(ASCP) Sales Manager, Histology Product Division Minnesota Medical Specialists, Inc. 7177 Madison Ave. W. Golden Valley, MN 55427-3601 763-542-8725 phone 888-790-9686 toll free 763-546-4830 fax email web _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. From TJJ <@t> Stowers-Institute.org Mon Jan 23 12:58:50 2006 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Mon Jan 23 13:16:20 2006 Subject: [Histonet] BrdU on fixed frozen section Message-ID: Eva, Are you denaturing them in HCl prior to BrdU antibody? This antibody requires denaturing by HCl or microwaving in order to recognize the BrdU epitope. DNA in cultured cells and frozen sections should be denatured in 2-4M HCl treatment, rinsed, and then incubated in antibody. If you are currently doing this and still get no signal, send me your protocol (including dilutions) and we can go from there. (Direct detection vs. indirect detection) How long are your samples fixed prior to freezing? Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From MVaughan4 <@t> ucok.edu Mon Jan 23 13:57:33 2006 From: MVaughan4 <@t> ucok.edu (MVaughan4@ucok.edu) Date: Mon Jan 23 13:57:59 2006 Subject: [Histonet] BrdU Staining HCl pretreat In-Reply-To: Message-ID: Eva, Did you pretreat the sample with 2N or 4N HCl? Mel Melville B. Vaughan, Ph. D. Assistant Professor Department of Biology University of Central Oklahoma 100 N. University Drive Edmond, OK 73034 I am trying to do a BrdU staining on frozen tissue slices (already fixed with PFA 4%) for weeks. The signal is restricted to the rim of the core and not within it. I am staining with anti-BrdU by the Roche detection kit. I already tried their protocol but that did not work on my fixed slices. I dry the slices then rehydrate for 1 h. After that I apply the 1st antibody for 30 min 37?C. Thanks for support. Eva Machen Sie aus 14 Cent spielend bis zu 100 Euro! Die neue Gaming-Area von Arcor - ?ber 50 Onlinespiele im Angebot. http://www.arcor.de/rd/emf-gaming-1 From Malcolm.McCallum <@t> tamut.edu Mon Jan 23 14:06:01 2006 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Mon Jan 23 14:06:22 2006 Subject: [Histonet] sliding microtome vs rotary microtome Message-ID: Hi, I am planning some skeletochronology and we have a sliding microtome. I have always used a rotary microtome and am curious if there are any tips from the pros regarding its utility for this purpose. I haven't used a sliding microtome since I was an undergrad. Malcolm McCallum From YuJ2 <@t> upmc.edu Mon Jan 23 15:11:28 2006 From: YuJ2 <@t> upmc.edu (Yu, Jian) Date: Mon Jan 23 15:11:38 2006 Subject: [Histonet] How to get good cross sections of the mouse small intestine Message-ID: <2554B4CA518D504A81E6908E39478A6A0D3CDF92@1upmc-msx10.isdip.upmc.edu> Dear all, I am a new histoneter. I am just starting to do some mouse small intestine histopathology and wondering whether anyone can provide a method (protocol) for how to fix and embed mouse small intestine to generate good cross (paraffin) sections with as many intact villi and crypts possible for H&E and IHC Particularly, I need to evaluate at least 5-10 different area of the small intestine to score the regeneration of crypts and the extend of apoptosis at various time point after irradiation. I have used neutral buffered formalin as fixative and manually bundle four pieces of 0.5 cm-long tubes together before embedding at a clinical pathology lab. The sections looked OK, but many appear to have a lot of floating villi in the lumen and limited number of crypts extending into to the villi connected to them. Since we can not always get our tissues processed after O/N fixation, we sometimes keep the tissues in NBF for several weeks. Is this too long, or should we transfer them to 70% ethanol? Look forward to your expert suggestions. With many thanks, ******************************************************** Jian Yu, Ph. D. University of Pittsburgh Cancer Institute Hillman Cancer Center Research Pavilion Office suite 2.26h, Laboratory 2.43 5117 Centre Avenue, Pittsburgh, PA 15213 Phone : 412-623-7786, (Lab) 412-623-3255 Fax: 412-623-7778 Email: yuj2@upmc.edu ******************************************************** From ana.merino-trigo <@t> wanadoo.fr Mon Jan 23 15:20:03 2006 From: ana.merino-trigo <@t> wanadoo.fr (ana.merino-trigo) Date: Mon Jan 23 15:20:28 2006 Subject: [Histonet] Embryons fixation and processing References: <20060123175037.82001.qmail@web61216.mail.yahoo.com> Message-ID: <031e01c62062$c6872fb0$5b790a0a@BABA> Hello everybody, I just began a new job in a different field. Before I was working with mouse xenograft tumors and now I moved to develpment biology field. I need to process zebrafish, chick, axolt and mouse embryos and I was wondering if anyone could advice in terms of fixation and processing times. Mostly of the papers that I got they fix in 4%PFA (I'm use to fix in 10% formalin, and I was wondering if is common with embryos to fix with 4%PFA). In some papers they fix 2 h at RT or 4?C o/n, when using 4%PFA. Any advice in this sense? In terms of processing, a guy that did some immunos with embryos here deshidrates samples in MeOH (25%, 50%, 75% and 100%, 5 min each), but I'm use to EtOH deshydratation (1x1h 70%; 2x1h 95%; 4x1 h 100%). Could someone advice in deshydratation times and advantages of using MeOH? Thanks a lot, Ana From RCazares <@t> schosp.org Mon Jan 23 15:34:27 2006 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Mon Jan 23 15:34:39 2006 Subject: [Histonet] H.pylori immunos on gastric biopsies Message-ID: <913FAC2B773C19488E26AE6572180FA50458A195@exch01.schosp.org> Hello Histonetters, I was wondering if anyone out there is doing immuno staining routinely on all gastric bx's? We are considering this in my lab, but I have my reservations. If some of you do, how many a week? And those who don't, why not? I will be doing a literature search on this topic, but if anyone knows of a specific article I sure would appreciate it if you could share it with me. Thank you, Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. From jim.manavis <@t> imvs.sa.gov.au Mon Jan 23 15:53:42 2006 From: jim.manavis <@t> imvs.sa.gov.au (Jim Manavis) Date: Mon Jan 23 15:53:53 2006 Subject: [Histonet] Human specific markers Message-ID: <000701c62067$7a1dc710$011e0a0a@itp36533> Can anyone suggest a human specific marker for use in Immnohistochemistry to distinguish between human and mouse cells Jim From Jackie.O'Connor <@t> abbott.com Mon Jan 23 16:03:02 2006 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Mon Jan 23 16:03:34 2006 Subject: [Histonet] Human specific markers Message-ID: Human mitochondria marker. I use it all the time. "Jim Manavis" Sent by: histonet-bounces@lists.utsouthwestern.edu 01/23/2006 03:53 PM To: "Histonet" cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Human specific markers Can anyone suggest a human specific marker for use in Immnohistochemistry to distinguish between human and mouse cells Jim _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon Jan 23 15:55:22 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Jan 23 16:34:46 2006 Subject: [Histonet] H.pylori immunos on gastric biopsies In-Reply-To: <913FAC2B773C19488E26AE6572180FA50458A195@exch01.schosp.org> Message-ID: <20060123215522.50894.qmail@web61215.mail.yahoo.com> Ruth: I always used a phosphotungstic modification of the Steiner procedure to avoid the radioactive uranyl nitrate [J.Histotechnology, 24(2):113-6; 2001) I never considered doing IHC because of costs. The Steiner procedure cost was $0.91/slide and any "typical" IHC procedure would cost about $8/slide if done manually and about $6/slide if done automatic at low capacity. For me those were good reasons not to use IHC for H. pilorii. Hope this will help you. Ren? J. "Cazares, Ruth" wrote: Hello Histonetters, I was wondering if anyone out there is doing immuno staining routinely on all gastric bx's? We are considering this in my lab, but I have my reservations. If some of you do, how many a week? And those who don't, why not? I will be doing a literature search on this topic, but if anyone knows of a specific article I sure would appreciate it if you could share it with me. Thank you, Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. From gcallis <@t> montana.edu Mon Jan 23 16:59:24 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Jan 23 16:59:37 2006 Subject: [Histonet] Re: How to get good cross sections of the mouse small intestine In-Reply-To: <2554B4CA518D504A81E6908E39478A6A0D3CDF92@1upmc-msx10.isdip .upmc.edu> References: <2554B4CA518D504A81E6908E39478A6A0D3CDF92@1upmc-msx10.isdip.upmc.edu> Message-ID: <6.0.0.22.1.20060123154522.01b2cec0@gemini.msu.montana.edu> Jian, You need to rinse the feces out of the lumen. Remove the small intestine INTACT but carefully by lifting it out of mouse. You can detach the mesentary as you do this but do not rip the sample out, you do NOT want to tear holes in the intestine. Fill a 50 ml syringe with PBS, and use a dulled 18 guage needle - insert into one end of the intestine (easier from the ilium side) and inject the PBS. After rinsing out feces, use a syringe filled with neutral buffered formalin, and inject that so the lumen distends. This will fix the villi much faster and get rid of digestive enzymes and left over fecal matter. You can clamp off the ends of the intestine or have sutures ready to tie off ends but you need to do this as the fixative is flowing out the opposite end of the intestine - overdistending the gut may not be a good idea, you need just enough to fix but not create a huge sausage like structure. Someone helping is nice, as they can tie off one end while the needle is removed to tie off the other end or use mosquito hemostats and clamp off. The idea is to fix the inside of intestine quickly to preserve villi. The gut can be stretched out in distended state or you can drop it into fixative then cut cross sections from each individual area of interest, process and embed on end. WE do this but NOT for cross sections, but mid sagittal sections however the intestine is well fixed, and with clean results. After fixation, you can store in 70% alcohol until processing. Just make sure you have total fixation or the 70% will finish it off for you. We are careful to not over process these tissues, they are small and delicate, and use a shorter processing schedule than for larger tissues. At 02:11 PM 1/23/2006, you wrote: >Dear all, > >I am a new histoneter. I am just starting to do some mouse small >intestine histopathology and wondering whether anyone can provide a >method (protocol) for how to fix and embed mouse small intestine to >generate good cross (paraffin) sections with as many intact villi and >crypts possible for H&E and IHC Particularly, I need to evaluate at >least 5-10 different area of the small intestine to score the >regeneration of crypts and the extend of apoptosis at various time point >after irradiation. > >I have used neutral buffered formalin as fixative and manually bundle >four pieces of 0.5 cm-long tubes together before embedding at a clinical >pathology lab. The sections looked OK, but many appear to have a lot of >floating villi in the lumen and limited number of crypts extending into >to the villi connected to them. Since we can not always get our >tissues processed after O/N fixation, we sometimes keep the tissues in >NBF for several weeks. Is this too long, or should we transfer them to >70% ethanol? Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From azdudley <@t> hotmail.com Mon Jan 23 17:22:21 2006 From: azdudley <@t> hotmail.com (anita dudley) Date: Mon Jan 23 17:22:36 2006 Subject: [Histonet] H.pylori immunos on gastric biopsies In-Reply-To: <913FAC2B773C19488E26AE6572180FA50458A195@exch01.schosp.org> Message-ID: ruth, we do immuno on all of our gastric bx's. we do them every day, the tech that comes in early cuts them and puts them in the 58 oven. the drs like them coming to them with the case. I was in a lecture at nat. meeting and the person said that the immuno stain was one of the best ways to demonstrate them. I could probably find the lecture for you in my national notebooks, if you like. anita dudley providence hosp mobile alabama >From: "Cazares, Ruth" >To: >Subject: [Histonet] H.pylori immunos on gastric biopsies >Date: Mon, 23 Jan 2006 15:34:27 -0600 > >Hello Histonetters, > > > >I was wondering if anyone out there is doing immuno staining routinely >on all gastric bx's? We are considering this in my lab, but I have my >reservations. If some of you do, how many a week? And those who don't, >why not? I will be doing a literature search on this topic, but if >anyone knows of a specific article I sure would appreciate it if you >could share it with me. > > > >Thank you, > > > >Ruth Cazares > > > > > >*** Confidentiality Statement *** >This e-mail is intended only for the use of the individual or entity to >which it is addressed and may contain information that is privileged and >confidential. If the reader of this message is not the intended recipient, >please notify the sender immediately by replying to this message and then >delete it from your system. Any review, dissemination, distribution, or >reproduction of this message by unintended recipients is strictly >prohibited and may be subject to legal restriction. > > >Thank you for your cooperation. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Mon Jan 23 17:44:44 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Jan 23 17:44:59 2006 Subject: [Histonet] How to get good cross sections of the mouse small intestine Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717654@lsexch.lsmaster.lifespan.org> In addition to Gayle's suggestions, you might want to physically hold the intestine straight while the formalin fixes and hardens it. After injecting formalin as Gayle said, I pin one end of it in a wax-lined dissection pan, then stretch it gently, just enough to straighten it, and pin the other end to keep it straight, then cover it with formalin. People sometimes send me intestine specimens which they have simply removed from the animal and dropped into formalin, resulting in a hardened gordian knot, all parts of which are curved to some degree, so that pieces cut from the intestine are shaped like parentheses. It's difficult to get a good perpendicular section of a curved object. If you don't have a dissecting pan you can easily make one; or you can just pin the specimen onto a strip of heavy corrugated cardbord or soft wood and drop it specimen side down into a dish of formalin. From igor.nasonkin <@t> jhmi.edu Mon Jan 23 18:40:50 2006 From: igor.nasonkin <@t> jhmi.edu (IGOR NASONKIN) Date: Mon Jan 23 18:41:00 2006 Subject: [Histonet] Human specific markers Message-ID: <3a4109cfaf1.43d51522@jhmimail.jhmi.edu> Jim, we use HNu (Human nuclei AB) from Chemicon; very robust, stains well and very specific. Staining is nuclear. Dr. Igor O. Nasonkin Ph.D. Postdoctoral Fellow Pathology/Neuropathology Johns Hopkins University Ross Research Bldg Rm 563 720 Rutland Ave Baltimore MD 21205 tel: 617-388-4104 Igor.Nasonkin@jhmi.edu ----- Original Message ----- From: Jim Manavis Date: Monday, January 23, 2006 2:53 pm Subject: [Histonet] Human specific markers > Can anyone suggest a human specific marker for use in > Immnohistochemistry to > distinguish between human and mouse cells > > Jim > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From glenn_krasinski <@t> yahoo.com Mon Jan 23 19:34:45 2006 From: glenn_krasinski <@t> yahoo.com (Glenn Krasinski) Date: Mon Jan 23 19:34:52 2006 Subject: [Histonet] Source for "Citrusolve"... Message-ID: <20060124013445.73779.qmail@web37107.mail.mud.yahoo.com> I am looking for a source for "citrusolve" for use instead of xylene in the deparaffinization of slides. Does it go by any other name? Many thanks. Glenn M. Krasinski San Diego, CA --------------------------------- Yahoo! Photos ? Showcase holiday pictures in hardcover Photo Books. You design it and we?ll bind it! From jqb7 <@t> cdc.gov Mon Jan 23 19:52:02 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Mon Jan 23 19:50:57 2006 Subject: [Histonet] Source for "Citrusolve"... References: <20060124013445.73779.qmail@web37107.mail.mud.yahoo.com> Message-ID: It's Citrisolve and Amity International carries it....probably some others do as well. http://ascenaa2.miniserver.com/~amity/products/citrisolve.htm Jeanine Bartlett CDC, Atlanta -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Glenn Krasinski Sent: Mon 1/23/2006 8:34 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Source for "Citrusolve"... I am looking for a source for "citrusolve" for use instead of xylene in the deparaffinization of slides. Does it go by any other name? Many thanks. Glenn M. Krasinski San Diego, CA --------------------------------- Yahoo! Photos ? Showcase holiday pictures in hardcover Photo Books. You design it and we?ll bind it! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eva-moeller <@t> arcor.de Tue Jan 24 03:07:50 2006 From: eva-moeller <@t> arcor.de (eva-moeller@arcor.de) Date: Tue Jan 24 03:08:02 2006 Subject: [Histonet] Aw: BrdU Staining HCl pretreat In-Reply-To: References: Message-ID: <4566989.1138093670423.JavaMail.ngmail@webmail-07.arcor-online.net> hello again, I forgot to mention that I cook the slices in citrate buffer for 5 min (800 Watts in microwave) before applying the antibody. I am now trying to do the HCl method instead. I will report my results. Thanks for helping me. Eva Eva Moeller Ruhr-Universitaet Bochum Germany Machen Sie aus 14 Cent spielend bis zu 100 Euro! Die neue Gaming-Area von Arcor - ?ber 50 Onlinespiele im Angebot. http://www.arcor.de/rd/emf-gaming-1 From juan.gutierrez <@t> christushealth.org Tue Jan 24 07:53:19 2006 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Tue Jan 24 07:53:29 2006 Subject: [Histonet] H.pylori immunos on gastric biopsies Message-ID: We do immunos on all gastric biopsies. We had some cases that looked negative by giemsa stain, yet the clinical hx suggested otherwise. When we ran the immunos on these so called negatives, we were shocked to see the amount of organisms we had missed. Do you want to take that chance? I always ask myself: what if this was my biopsy? Something to think about. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cazares, Ruth Sent: Monday, January 23, 2006 3:34 PM To: histonet@pathology.swmed.edu Subject: [Histonet] H.pylori immunos on gastric biopsies Hello Histonetters, I was wondering if anyone out there is doing immuno staining routinely on all gastric bx's? We are considering this in my lab, but I have my reservations. If some of you do, how many a week? And those who don't, why not? I will be doing a literature search on this topic, but if anyone knows of a specific article I sure would appreciate it if you could share it with me. Thank you, Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bsylinda <@t> aol.com Tue Jan 24 08:24:33 2006 From: bsylinda <@t> aol.com (bsylinda@aol.com) Date: Tue Jan 24 08:24:47 2006 Subject: [Histonet] CAP Question about Microwave Message-ID: <8C7EF090810EA74-968-20369@mblk-d37.sysops.aol.com> Hello Histoland, I am writing to ask if all labs are using lab approved microwaves and have them vented to an outside source. Are there still labs using microwaves from local retailers. If not how are you addressing the new CAp guidelines on checklist. Thanks in advance, Sylinda Battle, HT (ASCP) From twheelock <@t> mclean.harvard.edu Tue Jan 24 08:39:44 2006 From: twheelock <@t> mclean.harvard.edu (Tim Wheelock) Date: Tue Jan 24 08:41:56 2006 Subject: [Histonet] UN-SUBSCRIBE PLEASE Message-ID: <43D63C30.1060105@mclean.harvard.edu> I will be going on vacation. Please unsubscribe me. Thanks Any information, including protected health information (PHI), transmitted in this email is intended only for the person or entity to which it is addressed and may contain information that is privileged, confidential and or exempt from disclosure under applicable Federal or State law. Any review, retransmission, dissemination or other use of or taking of any action in reliance upon, protected health information (PHI) by persons or entities other than the intended recipient is prohibited. If you received this email in error, please contact the sender and delete the material from any computer. From Rodney.Slyter <@t> rivhs.com Tue Jan 24 09:05:28 2006 From: Rodney.Slyter <@t> rivhs.com (Slyter, Rodney) Date: Tue Jan 24 09:04:35 2006 Subject: [Histonet] job opening Message-ID: <95B1FC82E0F69D4FB1D2C04AAAF6CD721C7B1C@rivhsexchange.rhs.rivhs.local> There is an immediate opening for a histotech at riverside regional medical center in Newport news Virginia. This is a full-time morning position for a certified or eligible histotech. We provide a wide variety of tests including a wide battery of immunos, iSH and FISH. We do approx. 16,000 specimens a year both large resections and biopsies. There is no weekend or call coverage. There are two other full time techs, two full time lab aids, and a certified PA who is also a certified histotech. You can email me directly or visit the Riverside regional medical center website for more information. Rod Slyter, HTL(ASCP), PA(AAPA) Anatomical Pathology Manager Riverside Regional Medical Center 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 kjsavage <@t> buffalo.edu Tue Jan 24 09:06:52 2006 From: kjsavage <@t> buffalo.edu (kjsavage@buffalo.edu) Date: Tue Jan 24 09:07:06 2006 Subject: [Histonet] (no subject) Message-ID: <1138115212.43d6428c1aea3@mail3.buffalo.edu> Greetings all, I am having a difficult time trying to make methacarn fixative. I have searched the list archives and found a posting in May 2005 from someone that was having the same problem I am facing: a white precipitate forming. In one of the responses to his posting it was suggested to use glass only (pipets, vials etc...). I have done that, and still the precipitate forms. I have also used all new reagents (chloroform, methanol, acetic acid) and still the precipitate forms. Does anyone have some more suggestions as to what is going wrong? Thanks, Kathy Kathy Savage, PhD Dept. of Exercise and Nutrition Sciences SUNY-University at Buffalo From bill501 <@t> mindspring.com Tue Jan 24 09:29:19 2006 From: bill501 <@t> mindspring.com (Bill Blank) Date: Tue Jan 24 09:29:38 2006 Subject: [Histonet] H.pylori immunos on gastric biopsies In-Reply-To: References: Message-ID: IMO, we take this detecting H. pylori thing too seriously. A rational physician treats the patient and not any lab test. If I had symptoms of H. pylori, if my stomach looked inflamed, I would want to be treated for H. pylori irregardless of their presence on a biopsy. (Well, maybe as there is some evidence that treating antral H. pylori may increase the risk of GE junction adenocarcinoma) I consider immunos on gastric biopsies to be overkill and a waste of health care dollars. BIll At 7:53 AM -0600 1/24/06, GUTIERREZ, JUAN wrote: >We do immunos on all gastric biopsies. We had some cases that looked >negative by giemsa stain, yet the clinical hx suggested otherwise. When >we ran the immunos on these so called negatives, we were shocked to see >the amount of organisms we had missed. Do you want to take that chance? >I always ask myself: what if this was my biopsy? Something to think >about. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board From gcallis <@t> montana.edu Tue Jan 24 09:50:35 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Jan 24 09:50:43 2006 Subject: [Histonet] How to get good cross sections of the mouse small intestine In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE01717654@lsexch.lsmaster.l ifespan.org> References: <09C945920A6B654199F7A58A1D7D1FDE01717654@lsexch.lsmaster.lifespan.org> Message-ID: <6.0.0.22.1.20060124083902.01b439c8@gemini.msu.montana.edu> Paul, Excellent suggestion - for pinning down the samples, I suggest using disposable hypodermic needles - the latter do NOT rust or corrode in aqueous based NBF - needles are stainless steel. Tiny needles (24 or less gauge) for this purpose and not tear the ends of the intestine to bits with larger gauge. The Gordian knot description was so true and our loops of intestine always were filled with mouse feces!!! Once fixed, couldn't removed - microtomy was no fun and sections looked terrible. You can also use cork strips (purchased from hardware store, very cheap) then toss them out later. We would stand these on end in a deep bucket with lid - tagged carefully. And a cheap pan with a lid is the cake pan sizes or larger, with lids - Rubbermaid or some other plastic. Very nice to reach into a pan and get the samples - loved this idea. At 04:44 PM 1/23/2006, you wrote: > In addition to Gayle's suggestions, you might want to physically >hold the intestine straight while the formalin fixes and hardens it. After >injecting formalin as Gayle said, I pin one end of it in a wax-lined >dissection pan, then stretch it gently, just enough to straighten it, and >pin the other end to keep it straight, then cover it with formalin. People >sometimes send me intestine specimens which they have simply removed from >the animal and dropped into formalin, resulting in a hardened gordian knot, >all parts of which are curved to some degree, so that pieces cut from the >intestine are shaped like parentheses. It's difficult to get a good >perpendicular section of a curved object. If you don't have a dissecting >pan you can easily make one; or you can just pin the specimen onto a strip >of heavy corrugated cardbord or soft wood and drop it specimen side down >into a dish of formalin. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From bsylinda <@t> aol.com Tue Jan 24 09:57:06 2006 From: bsylinda <@t> aol.com (bsylinda@aol.com) Date: Tue Jan 24 09:57:22 2006 Subject: [Histonet] CAP Question about Microwave In-Reply-To: <8C7EF090810EA74-968-20369@mblk-d37.sysops.aol.com> References: <8C7EF090810EA74-968-20369@mblk-d37.sysops.aol.com> Message-ID: <8C7EF15F587F78E-A58-150A6@FWM-D06.sysops.aol.com> An additionan to comment. This microwave is only used for special stains no tissuse processing. Thanks again, sylinda -----Original Message----- From: bsylinda@aol.com To: histonet@lists.utsouthwestern.edu Sent: Tue, 24 Jan 2006 09:24:33 -0500 Subject: [Histonet] CAP Question about Microwave Hello Histoland, I am writing to ask if all labs are using lab approved microwaves and have them vented to an outside source. Are there still labs using microwaves from local retailers. If not how are you addressing the new CAp guidelines on checklist. Thanks in advance, Sylinda Battle, HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jeffery-Smith <@t> ouhsc.edu Tue Jan 24 10:17:45 2006 From: Jeffery-Smith <@t> ouhsc.edu (Smith, Jeffery D. (HSC)) Date: Tue Jan 24 10:20:06 2006 Subject: [Histonet] H.pylori immunos on gastric biopsies Message-ID: I have to somewhat agree. Immunos for H. pylori seems to be overkill. We routinely stain for H. pylori using a Giemsa on all GI biopsies. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bill Blank Sent: Tue 1/24/2006 9:29 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies IMO, we take this detecting H. pylori thing too seriously. A rational physician treats the patient and not any lab test. If I had symptoms of H. pylori, if my stomach looked inflamed, I would want to be treated for H. pylori irregardless of their presence on a biopsy. (Well, maybe as there is some evidence that treating antral H. pylori may increase the risk of GE junction adenocarcinoma) I consider immunos on gastric biopsies to be overkill and a waste of health care dollars. BIll At 7:53 AM -0600 1/24/06, GUTIERREZ, JUAN wrote: >We do immunos on all gastric biopsies. We had some cases that looked >negative by giemsa stain, yet the clinical hx suggested otherwise. When >we ran the immunos on these so called negatives, we were shocked to see >the amount of organisms we had missed. Do you want to take that chance? >I always ask myself: what if this was my biopsy? Something to think >about. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SBarnes <@t> elch.org Tue Jan 24 10:27:14 2006 From: SBarnes <@t> elch.org (Sue Barnes) Date: Tue Jan 24 10:27:22 2006 Subject: [Histonet] Health Testing when using xylene Message-ID: Does anyone in the histology world have health testing for techs who use xylene? What testing is done? Is there printed regulations for suggested health testing? Any help on this would be appreciated. Thanks Sue Barnes East Liverpool City Hospital East Liverpool, Ohio From pmcardle <@t> ebsciences.com Tue Jan 24 10:32:29 2006 From: pmcardle <@t> ebsciences.com (Phil McArdle) Date: Tue Jan 24 10:32:45 2006 Subject: [Histonet] CAP Question about Microwave Message-ID: <43D6569D.5010309@ebsciences.com> Hello: If you want some "vendor" input, please feel free to contact me. Best regards, Phil McArdle -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" From rjbuesa <@t> yahoo.com Tue Jan 24 10:48:28 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Jan 24 10:48:40 2006 Subject: [Histonet] Health Testing when using xylene In-Reply-To: Message-ID: <20060124164828.11569.qmail@web61214.mail.yahoo.com> Sue: I used to test for formaldehyde, glutaraldehyde, xylene and the air flow level of the fumes hoods. The fumes hoods had simple air gauges tha were checked daily and the results written in a log in the hood door. For the xylene (as well as the others) we used pasive monitoring badges worn during 8 hours (to determine Time Weighed Average or TWA) or during 15 minutes (to determine Short Term Exposure Level or STEL). Some of the badges were developed in our lab and others were sent to an outside laboratory for confirmation of results. After 8 years of cummulative data with values below OSHA levels, and because this was a very expensive program, we contracted an outside company that did the monitoring only in the areas and we stopped doing the personal monitoring. Hope this will help you!. Ren? J. Sue Barnes wrote: Does anyone in the histology world have health testing for techs who use xylene? What testing is done? Is there printed regulations for suggested health testing? Any help on this would be appreciated. Thanks Sue Barnes East Liverpool City Hospital East Liverpool, Ohio _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From RSRICHMOND <@t> aol.com Tue Jan 24 11:07:15 2006 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Tue Jan 24 11:07:25 2006 Subject: [Histonet] H. pylori immunos on gastric biopsies Message-ID: <228.5316898.3107b8c3@aol.com> I've finally had a chance to compare two methods - our lab changed from Giemsa (actually Diff-Quik II generic equivalent) to immunostaining a few months ago. It's a lot easier to find the bacteria with the immunostain than it is with the dye technique, but bacterial morphology is not as good. If I were seeing 17 total cases a day with one gastric biopsy every other day, and had time to search the slide with an oil immersion lens, I'd rather have the dye technique. But when I have to sign out 60 cases in a day, with 10 gastric biopsies, I'd rather have the immunostain. Clinically, there are several ways to make the diagnosis: biopsy sections, urease (Clo-Test) testing of a biopsy specimens, serology, isotopically labeled urea with breath testing. Obviously as a pathologist I have my prejudices, but there's nothing like tissue sections to pin down the diagnosis of chronic active gastritis and make sure the patient doesn't have a lymphoma or carcinoma lurking in there as well. Bob Richmond Gaston Memorial Hospital, Gastonia NC From Terry.Marshall <@t> rothgen.nhs.uk Tue Jan 24 11:09:21 2006 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Tue Jan 24 11:08:06 2006 Subject: [Histonet] H.pylori immunos on gastric biopsies Message-ID: H. pylori is a pathogen. It does not sit in the stomach behaving itself. Its presence is an indication for treatment, but its absence is *not* an indication for treatment directed against it, irrespective of how inflamed the stomach looks. The best histologic determinant of H. pylori is an immunostain. However, in most cases, this is all rather academic, as a quick stool test is available, directed against H. pylori antigen, so in most cases, we are really trying to determine something better done by other means. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Smith, Jeffery D. (HSC) [mailto:Jeffery-Smith@ouhsc.edu] Sent: 24 January 2006 16:18 To: Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies I have to somewhat agree. Immunos for H. pylori seems to be overkill. We routinely stain for H. pylori using a Giemsa on all GI biopsies. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bill Blank Sent: Tue 1/24/2006 9:29 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies IMO, we take this detecting H. pylori thing too seriously. A rational physician treats the patient and not any lab test. If I had symptoms of H. pylori, if my stomach looked inflamed, I would want to be treated for H. pylori irregardless of their presence on a biopsy. (Well, maybe as there is some evidence that treating antral H. pylori may increase the risk of GE junction adenocarcinoma) I consider immunos on gastric biopsies to be overkill and a waste of health care dollars. BIll At 7:53 AM -0600 1/24/06, GUTIERREZ, JUAN wrote: >We do immunos on all gastric biopsies. We had some cases that looked >negative by giemsa stain, yet the clinical hx suggested otherwise. When >we ran the immunos on these so called negatives, we were shocked to see >the amount of organisms we had missed. Do you want to take that chance? >I always ask myself: what if this was my biopsy? Something to think >about. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@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 ewrona <@t> yahoo.com Tue Jan 24 12:02:59 2006 From: ewrona <@t> yahoo.com (Erin Wrona) Date: Tue Jan 24 12:03:07 2006 Subject: [Histonet] Bone marrow enzyme stains Message-ID: <20060124180259.78156.qmail@web52514.mail.yahoo.com> Hi All, About two years ago we were told that our supplier would no longer be manufacturing the kit for beta-Naphthyl Butyrate Esterase Lymphocyte. We started using Alpha-Naphthyl Acetate Esterase with and without flouride as a "temporary back up". Our bone marrow pathologists would like to go back to the beta. Does anyone know of a supplier, or something comparable? What do you use in your facility? Thanks so much, Erin Wrona San Francisco From weneng2004 <@t> yahoo.com Tue Jan 24 12:36:19 2006 From: weneng2004 <@t> yahoo.com (wen eng) Date: Tue Jan 24 12:36:32 2006 Subject: [Histonet] Perls' Prussian Iron Stain Message-ID: <20060124183619.53310.qmail@web53407.mail.yahoo.com> Hello histonetters, I did Perls' stain on mouse liver. I undersstand that the bright blue is the target I should focus on. But I also see a lot dark brown stain on them which I don't know what it is. I am eager to know because I do see big difference between treated and untreated animals. I checked my text book and other resouces but could not find the answer. I hope the histonetters here can kindly help me with this. Thanks! Wen --------------------------------- What are the most popular cars? Find out at Yahoo! Autos From juan.gutierrez <@t> christushealth.org Tue Jan 24 12:57:24 2006 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Tue Jan 24 12:57:33 2006 Subject: [Histonet] p57 antibody Message-ID: Hi all! Is anybody using this ab? My pathologist wants me to work it out, but all I've been able to find are RUO's. Is there a source for IVD's? We are not interested in sending the cases out to reference labs so please don't ask. Thanks for your help. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! From sluhisto <@t> yahoo.com Tue Jan 24 13:13:48 2006 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Tue Jan 24 13:13:56 2006 Subject: [Histonet] Histology Salaries Message-ID: <20060124191348.22483.qmail@web51009.mail.yahoo.com> Hello All: I am really in need of having our salaries evaluated. Unfortunately, to get anyones attention, I need to have some starting information. Would any of you mind sharing, confidentially, your salary structure for HT, HTL, CT, and Histology Assistants? I promise not to identify your institution in my report to my manager if you wish. As a matter of fact, you can respond with just the area of the country if you wish. Any help you can give would be appreciated. I realize that there is information available from NSH or possibly CAP but it is always behind and I would really like to hear directly from you guys. Thanks, in advance. Susan --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. From katherine_dubrall <@t> yahoo.com Tue Jan 24 13:22:33 2006 From: katherine_dubrall <@t> yahoo.com (kay dubrall) Date: Tue Jan 24 13:22:43 2006 Subject: [Histonet] switching email Message-ID: <20060124192233.70538.qmail@web54215.mail.yahoo.com> can i stop histonet email to one email account and switch them to another in one swoop? what's kay is kay - j3 --------------------------------- Yahoo! Photos Ring in the New Year with Photo Calendars. Add photos, events, holidays, whatever. From SBarnes <@t> elch.org Tue Jan 24 13:39:37 2006 From: SBarnes <@t> elch.org (Sue Barnes) Date: Tue Jan 24 13:39:47 2006 Subject: [Histonet] ribbon pro (fisher paraffin) Message-ID: Is anyone using the ribbon pro from Fisher Scientific? If so are you having any problems with it. We are having problems with air pockets and chatter with it. When using a control block embedded previously there is no problem so it doesn't appear to be the microtome, knife or water bath. I have had problems with the ribbon pro for a while now, seeing things that I have never seen before. Is there someone out there who can help? Thanks Sue From dsantana <@t> pmaonline.com Tue Jan 24 14:10:05 2006 From: dsantana <@t> pmaonline.com (Santana, Diane) Date: Tue Jan 24 14:09:40 2006 Subject: [Histonet] pas Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB071137FB@MAILPMA> Hi, I am just curious on how others are doing a PAS for fungus? Thanks Diane Santana From Sjohnso616 <@t> aol.com Tue Jan 24 14:27:13 2006 From: Sjohnso616 <@t> aol.com (Sjohnso616@aol.com) Date: Tue Jan 24 14:27:42 2006 Subject: [Histonet] Job opportunity in Florida Message-ID: <1c5.38d3f250.3107e7a1@aol.com> Hi Histonetters, Sarasota Pathology in Sarasota Florida has openings for ASCP HT. If you are looking for a place that stresses teamwork, offers great benefits, and offers a marvelous climate please send resume to _sjohnson@sarapath.com_ (mailto:sjohnson@sarapath.com) . or _nday@sarapath.com_ (mailto:nday@sarapath.com) Thank you Saundra Johnson Sarasota Pathology Histology-IHC Supervisor From PMonfils <@t> Lifespan.org Tue Jan 24 15:11:55 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Tue Jan 24 15:12:07 2006 Subject: [Histonet] pas Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717657@lsexch.lsmaster.lifespan.org> I have used a modified Gridley technique for many years to stain fungi. It uses a stronger oxidizer (chromic acid instead of periodic acid), which tends to over-oxidize tissue components like glycogen, mucin, elastin, fibrin, reticulin, basement membranes, etc., so that they no longer react with the Schiff Reagent, leaving only the fungal structures to react. It is therefore more specific for fungi than ordinary PAS procedures. The original Gridley technique also employs aldehyde fuchsin to enhance the stain; however I usually omit this step because aldehyde fuchsin stains elastin and some other tissue components, thereby partially negating the advantage of using chromic acid. Particularly in tissues with abundant elastin, like skin and lung, using aldehyde fuchsin can make it more difficult to identify fungal hyphae, especially when they are sparse. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Santana, Diane > Sent: Tuesday, January 24, 2006 12:10 PM > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] pas > > Hi, > I am just curious on how others are doing a PAS for fungus? > Thanks > Diane Santana > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Jeffery-Smith <@t> ouhsc.edu Tue Jan 24 15:18:16 2006 From: Jeffery-Smith <@t> ouhsc.edu (Smith, Jeffery D. (HSC)) Date: Tue Jan 24 15:20:35 2006 Subject: [Histonet] pas Message-ID: I run a routine PAS just changing the counterstain from hematoxylin to Light Green. The green masks most of the other components leaving bright pink fungi on a pink background. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Santana, Diane Sent: Tue 1/24/2006 2:10 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] pas Hi, I am just curious on how others are doing a PAS for fungus? Thanks Diane Santana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Tue Jan 24 15:22:52 2006 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Tue Jan 24 15:23:06 2006 Subject: AW: [Histonet] Health Testing when using xylene In-Reply-To: Message-ID: <000001c6212c$58f581c0$eeeea8c0@SERVER01> In Austria we have a law for testing methylhippuracid in urin of people, that are in contact with xylol, twice in the year. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Sue Barnes Gesendet: Dienstag, 24. J?nner 2006 17:27 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Health Testing when using xylene Does anyone in the histology world have health testing for techs who use xylene? What testing is done? Is there printed regulations for suggested health testing? Any help on this would be appreciated. Thanks Sue Barnes East Liverpool City Hospital East Liverpool, Ohio _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jeffery-Smith <@t> ouhsc.edu Tue Jan 24 15:25:33 2006 From: Jeffery-Smith <@t> ouhsc.edu (Smith, Jeffery D. (HSC)) Date: Tue Jan 24 15:26:32 2006 Subject: [Histonet] pas Message-ID: Sorry I got distracted that was pink fungi on green background. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Smith, Jeffery D. (HSC) Sent: Tue 1/24/2006 3:18 PM To: Santana, Diane; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] pas I run a routine PAS just changing the counterstain from hematoxylin to Light Green. The green masks most of the other components leaving bright pink fungi on a pink background. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Santana, Diane Sent: Tue 1/24/2006 2:10 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] pas Hi, I am just curious on how others are doing a PAS for fungus? Thanks Diane Santana _______________________________________________ Histonet mailing list Histonet@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 Jeffery-Smith <@t> ouhsc.edu Tue Jan 24 16:28:23 2006 From: Jeffery-Smith <@t> ouhsc.edu (Smith, Jeffery D. (HSC)) Date: Tue Jan 24 16:32:50 2006 Subject: [Histonet] pas Message-ID: Stock Light Green Light Green 1gm Distilled water 500 mL Glacial Acetic Acid 1mL Mix well and filter. Stock solution stable for 3 to 4 months. Working Light Green Stock light green 10mL Distilled Water 50mL ________________________________ From: BennettW@pac.dfo-mpo.gc.ca [mailto:BennettW@pac.dfo-mpo.gc.ca] Sent: Tue 1/24/2006 3:55 PM To: Smith, Jeffery D. (HSC) Subject: RE: [Histonet] pas Hi Diane, What concentration of Light Green did you use? Cheers Bill Bennett Histologist Fisheries and Oceans Canada Pacific Biological Station -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Smith, Jeffery D. (HSC) Sent: Tuesday, January 24, 2006 1:18 PM To: Santana, Diane; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] pas I run a routine PAS just changing the counterstain from hematoxylin to Light Green. The green masks most of the other components leaving bright pink fungi on a pink background. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Santana, Diane Sent: Tue 1/24/2006 2:10 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] pas Hi, I am just curious on how others are doing a PAS for fungus? Thanks Diane Santana _______________________________________________ Histonet mailing list Histonet@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 Tue Jan 24 17:12:50 2006 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Tue Jan 24 17:13:03 2006 Subject: [Histonet] Zinc Formalin Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628006307C8B@EXCHANGE1.huntingtonhospital.com> Our formalin supplier mistakenly shipped us pre-filled containers containing zinc formalin instead of 10% NBF. Before I noticed this, three cases had been distributed throughout the hospital (most likely to Surgery). So, how is this going to affect my tissue? Is there any post-treatment (such as removing mercury pigment on B-5 fixed tissue) that I need to perform? Thanks in advance for any info you can provide. Laurie Colbert From glenn_krasinski <@t> yahoo.com Tue Jan 24 17:05:03 2006 From: glenn_krasinski <@t> yahoo.com (Glenn Krasinski) Date: Tue Jan 24 17:32:15 2006 Subject: [Histonet] Need for antigen retrieval question... Message-ID: <20060124230503.59072.qmail@web37111.mail.mud.yahoo.com> My boss will be injecting mice burdened with solid tumors with antibodies specific to potential biomarkers. The tumors/tissues from the sacrificed animals will be fixed in formalin and embedded in paraffin blocks. Sections will then be stained with secondary antibodies to the specific biomarker antibodies. My question is whether or not I will need to go through an antigen retrieval process. My apologies for the complicated question. Glenn M. Krasinski San Diego, CA 92121 --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. From histology.bc <@t> shaw.ca Tue Jan 24 17:55:34 2006 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Tue Jan 24 17:55:44 2006 Subject: [Histonet] Perls' Prussian Iron Stain In-Reply-To: <20060124183619.53310.qmail@web53407.mail.yahoo.com> References: <20060124183619.53310.qmail@web53407.mail.yahoo.com> Message-ID: <43D6BE76.5010202@shaw.ca> > Without seeing the brown pigment it is hard to say what it is. But, if > you are dealing with sections of mouse liver, my first guess would be > that the brown material is bile pigment. You are correct in assuming > that the blue material is the product formed by the reaction of ferric > iron with Perls' reagents. You say that you see a big difference between treated and untreated animals. What sort of treatment have they had? Let us have some more information, and maybe we can help some more. Paul Kamloops, BC Canada ********************************************************************* > > I did Perls' stain on mouse liver. I undersstand that the bright blue is the target I should focus on. But I also see a lot dark brown stain on them which I don't know what it is. I am eager to know because I do see big difference between treated and untreated animals. I checked my text book and other resouces but could not find the answer. I hope the histonetters here can kindly help me with this. > > Thanks! > > Wen > > >--------------------------------- > > What are the most popular cars? Find out at Yahoo! Autos >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From ylee <@t> bccancer.bc.ca Tue Jan 24 19:25:23 2006 From: ylee <@t> bccancer.bc.ca (Lee, Yin Ping) Date: Tue Jan 24 19:25:33 2006 Subject: [Histonet] MDM2 Message-ID: <57AD92D1EDD1854BAAC02DD7DDAF0E360172E010@srvex10.phsabc.ehcnet.ca> Hi, I'm trying Dako MDM2 clone SMP14 and Vector MDM2 clone 1B10. They both give me weak staining with dirty background. Does anyone have any experience with this antibody: such as which clone (and supplier) is better or tips regarding the technical details (antigen retrieval, dilution etc). Thanks in advance for any help. Yin Ping Lee Histopathology BC Cancer Agency Vancouver BC, Canada From fadiafandi <@t> mac.com Tue Jan 24 21:10:27 2006 From: fadiafandi <@t> mac.com (fadiafandi@mac.com) Date: Tue Jan 24 21:10:44 2006 Subject: [Histonet] Re: H. Pylori work-up In-Reply-To: <200601241810.k0OIA3am014726@smtpout.mac.com> References: <200601241810.k0OIA3am014726@smtpout.mac.com> Message-ID: <303988A5-99A3-4D08-B280-22FCF6CAE88B@mac.com> Regarding H. Pylori: It may be wise to use Steiner (instead of Giemsa) as a favored tissue- based screening method for H. Pylori. In our experience, about 5-10% of cases stained with Steiner end up being morphologically equivocal where you view the organism but it doesn't have definitive morphologic features of HP or HH. In those cases, we defer to IHC for confirmation, which is the definitive tissue-based test. But I don't think IHC should be run as a screening tool. In terms of cost differences, a published article by Tulaymat et al. in the Archives (I believe from 1999) actually shows that IHC is more cost-effective. You could purchase a concentrated antibody and dilute it out. In that case, the cost difference between silver stains an IHC may not be that drastic. Best, Hadi Yaziji, M.D. On Jan 24, 2006, at 1:10 PM, histonet- request@lists.utsouthwestern.edu wrote: > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. BrdU on fixed frozen section (Johnson, Teri) > 2. BrdU Staining HCl pretreat (MVaughan4@ucok.edu) > 3. sliding microtome vs rotary microtome (Malcolm McCallum) > 4. How to get good cross sections of the mouse small intestine > (Yu, Jian) > 5. Embryons fixation and processing (ana.merino-trigo) > 6. H.pylori immunos on gastric biopsies (Cazares, Ruth) > 7. Human specific markers (Jim Manavis) > 8. Re: Human specific markers (Jackie M O'Connor) > 9. Re: H.pylori immunos on gastric biopsies (Rene J Buesa) > 10. Re: How to get good cross sections of the mouse small > intestine (Gayle Callis) > 11. RE: H.pylori immunos on gastric biopsies (anita dudley) > 12. RE: How to get good cross sections of the mouse small > intestine (Monfils, Paul) > 13. Re: Human specific markers (IGOR NASONKIN) > 14. Source for "Citrusolve"... (Glenn Krasinski) > 15. RE: Source for "Citrusolve"... (Bartlett, Jeanine) > 16. Aw: BrdU Staining HCl pretreat (eva-moeller@arcor.de) > 17. RE: H.pylori immunos on gastric biopsies (GUTIERREZ, JUAN) > 18. CAP Question about Microwave (bsylinda@aol.com) > 19. UN-SUBSCRIBE PLEASE (Tim Wheelock) > 20. job opening (Slyter, Rodney) > 21. (no subject) (kjsavage@buffalo.edu) > 22. RE: H.pylori immunos on gastric biopsies (Bill Blank) > 23. RE: How to get good cross sections of the mouse small > intestine (Gayle Callis) > 24. Re: CAP Question about Microwave (bsylinda@aol.com) > 25. RE: H.pylori immunos on gastric biopsies > (Smith, Jeffery D. (HSC)) > 26. Health Testing when using xylene (Sue Barnes) > 27. Re: CAP Question about Microwave (Phil McArdle) > 28. Re: Health Testing when using xylene (Rene J Buesa) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Mon, 23 Jan 2006 12:58:50 -0600 > From: "Johnson, Teri" > Subject: [Histonet] BrdU on fixed frozen section > To: > Cc: histonet@lists.utsouthwestern.edu > Message-ID: > institute.org> > > Content-Type: text/plain; charset="us-ascii" > > Eva, > > Are you denaturing them in HCl prior to BrdU antibody? This antibody > requires denaturing by HCl or microwaving in order to recognize the > BrdU > epitope. DNA in cultured cells and frozen sections should be denatured > in 2-4M HCl treatment, rinsed, and then incubated in antibody. > > If you are currently doing this and still get no signal, send me your > protocol (including dilutions) and we can go from there. (Direct > detection vs. indirect detection) How long are your samples fixed > prior > to freezing? > > Teri Johnson, HT(ASCP)QIHC > Managing Director Histology Facility > Stowers Institute for Medical Research > 1000 E. 50th St. > Kansas City, MO 64133 > > > > ------------------------------ > > Message: 2 > Date: Mon, 23 Jan 2006 13:57:33 -0600 > From: MVaughan4@ucok.edu > Subject: [Histonet] BrdU Staining HCl pretreat > To: histonet@lists.utsouthwestern.edu > Cc: eva-moeller@arcor.de > Message-ID: > > Content-Type: text/plain; charset="ISO-8859-1" > > Eva, > Did you pretreat the sample with 2N or 4N HCl? > > Mel > Melville B. Vaughan, Ph. D. > Assistant Professor > Department of Biology > University of Central Oklahoma > 100 N. University Drive > Edmond, OK 73034 > > I am trying to do a BrdU staining on frozen tissue slices (already > fixed > with PFA 4%) for weeks. > The signal is restricted to the rim of the core and not within it. > I am > staining with anti-BrdU by the Roche detection kit. I already tried > their > protocol but that did not work on my fixed slices. > I dry the slices then rehydrate for 1 h. After that I apply the 1st > antibody for 30 min 37?C. > Thanks for support. > Eva > > Machen Sie aus 14 Cent spielend bis zu 100 Euro! > Die neue Gaming-Area von Arcor - ?ber 50 Onlinespiele im Angebot. > http://www.arcor.de/rd/emf-gaming-1 > > > > ------------------------------ > > Message: 3 > Date: Mon, 23 Jan 2006 14:06:01 -0600 > From: "Malcolm McCallum" > Subject: [Histonet] sliding microtome vs rotary microtome > To: > Message-ID: > > Content-Type: text/plain; charset="us-ascii" > > Hi, > > I am planning some skeletochronology and we have a sliding > microtome. I > have always used a rotary microtome and am curious if there are any > tips > from the pros regarding its utility for this purpose. > > > > I haven't used a sliding microtome since I was an undergrad. > > > > Malcolm McCallum > > > > ------------------------------ > > Message: 4 > Date: Mon, 23 Jan 2006 16:11:28 -0500 > From: "Yu, Jian" > Subject: [Histonet] How to get good cross sections of the mouse small > intestine > To: > Message-ID: > <2554B4CA518D504A81E6908E39478A6A0D3CDF92@1upmc-msx10.isdip.upmc.edu> > Content-Type: text/plain; charset="us-ascii" > > Dear all, > > I am a new histoneter. I am just starting to do some mouse small > intestine histopathology and wondering whether anyone can provide a > method (protocol) for how to fix and embed mouse small intestine to > generate good cross (paraffin) sections with as many intact villi and > crypts possible for H&E and IHC Particularly, I need to evaluate at > least 5-10 different area of the small intestine to score the > regeneration of crypts and the extend of apoptosis at various time > point > after irradiation. > > I have used neutral buffered formalin as fixative and manually bundle > four pieces of 0.5 cm-long tubes together before embedding at a > clinical > pathology lab. The sections looked OK, but many appear to have a > lot of > floating villi in the lumen and limited number of crypts extending > into > to the villi connected to them. Since we can not always get our > tissues processed after O/N fixation, we sometimes keep the tissues in > NBF for several weeks. Is this too long, or should we transfer > them to > 70% ethanol? > > > > Look forward to your expert suggestions. > > With many thanks, > > ******************************************************** > > Jian Yu, Ph. D. > > University of Pittsburgh Cancer Institute > > Hillman Cancer Center Research Pavilion > > Office suite 2.26h, Laboratory 2.43 > > 5117 Centre Avenue, Pittsburgh, PA 15213 > > > > Phone : 412-623-7786, (Lab) 412-623-3255 > > Fax: 412-623-7778 > > Email: yuj2@upmc.edu > > ******************************************************** > > > > > > ------------------------------ > > Message: 5 > Date: Mon, 23 Jan 2006 13:20:03 -0800 > From: "ana.merino-trigo" > Subject: [Histonet] Embryons fixation and processing > To: "Histonet" > Message-ID: <031e01c62062$c6872fb0$5b790a0a@BABA> > Content-Type: text/plain; charset="iso-8859-1" > > Hello everybody, > > I just began a new job in a different field. Before I was working > with mouse xenograft tumors and now I moved to develpment biology > field. I need to process zebrafish, chick, axolt and mouse embryos > and I was wondering if anyone could advice in terms of fixation and > processing times. Mostly of the papers that I got they fix in 4%PFA > (I'm use to fix in 10% formalin, and I was wondering if is common > with embryos to fix with 4%PFA). In some papers they fix 2 h at RT > or 4?C o/n, when using 4%PFA. Any advice in this sense? > > In terms of processing, a guy that did some immunos with embryos > here deshidrates samples in MeOH (25%, 50%, 75% and 100%, 5 min > each), but I'm use to EtOH deshydratation (1x1h 70%; 2x1h 95%; 4x1 > h 100%). Could someone advice in deshydratation times and > advantages of using MeOH? > > Thanks a lot, > > Ana > > > ------------------------------ > > Message: 6 > Date: Mon, 23 Jan 2006 15:34:27 -0600 > From: "Cazares, Ruth" > Subject: [Histonet] H.pylori immunos on gastric biopsies > To: > Message-ID: > <913FAC2B773C19488E26AE6572180FA50458A195@exch01.schosp.org> > Content-Type: text/plain; charset="us-ascii" > > Hello Histonetters, > > > > I was wondering if anyone out there is doing immuno staining routinely > on all gastric bx's? We are considering this in my lab, but I have my > reservations. If some of you do, how many a week? And those who > don't, > why not? I will be doing a literature search on this topic, but if > anyone knows of a specific article I sure would appreciate it if you > could share it with me. > > > > Thank you, > > > > Ruth Cazares > > > > > > *** Confidentiality Statement *** > This e-mail is intended only for the use of the individual or > entity to which it is addressed and may contain information that is > privileged and confidential. If the reader of this message is not > the intended recipient, please notify the sender immediately by > replying to this message and then delete it from your system. Any > review, dissemination, distribution, or reproduction of this > message by unintended recipients is strictly prohibited and may be > subject to legal restriction. > > > Thank you for your cooperation. > > > > ------------------------------ > > Message: 7 > Date: Tue, 24 Jan 2006 08:23:42 +1030 > From: "Jim Manavis" > Subject: [Histonet] Human specific markers > To: "Histonet" > Message-ID: <000701c62067$7a1dc710$011e0a0a@itp36533> > Content-Type: text/plain; charset="iso-8859-1" > > Can anyone suggest a human specific marker for use in > Immnohistochemistry to > distinguish between human and mouse cells > > Jim > > > > > ------------------------------ > > Message: 8 > Date: Mon, 23 Jan 2006 16:03:02 -0600 > From: "Jackie M O'Connor" > Subject: Re: [Histonet] Human specific markers > To: "Jim Manavis" > Cc: Histonet > Message-ID: > Content-Type: text/plain; charset="us-ascii" > > Human mitochondria marker. I use it all the time. > > > > > > > "Jim Manavis" > Sent by: histonet-bounces@lists.utsouthwestern.edu > 01/23/2006 03:53 PM > > > To: "Histonet" > cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) > Subject: [Histonet] Human specific markers > > > Can anyone suggest a human specific marker for use in > Immnohistochemistry > to > distinguish between human and mouse cells > > Jim > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 9 > Date: Mon, 23 Jan 2006 13:55:22 -0800 (PST) > From: Rene J Buesa > Subject: Re: [Histonet] H.pylori immunos on gastric biopsies > To: "Cazares, Ruth" , > histonet@pathology.swmed.edu > Message-ID: <20060123215522.50894.qmail@web61215.mail.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > Ruth: > I always used a phosphotungstic modification of the Steiner > procedure to avoid > the radioactive uranyl nitrate [J.Histotechnology, 24(2):113-6; > 2001) > I never considered doing IHC because of costs. > The Steiner procedure cost was $0.91/slide and any "typical" IHC > procedure > would cost about $8/slide if done manually and about $6/slide if > done automatic > at low capacity. > For me those were good reasons not to use IHC for H. pilorii. > Hope this will help you. > Ren? J. > > "Cazares, Ruth" wrote: > Hello Histonetters, > > > > I was wondering if anyone out there is doing immuno staining routinely > on all gastric bx's? We are considering this in my lab, but I have my > reservations. If some of you do, how many a week? And those who don't, > why not? I will be doing a literature search on this topic, but if > anyone knows of a specific article I sure would appreciate it if you > could share it with me. > > > > Thank you, > > > > Ruth Cazares > > > > > > *** Confidentiality Statement *** > This e-mail is intended only for the use of the individual or > entity to which it is addressed and may contain information that is > privileged and confidential. If the reader of this message is not > the intended recipient, please notify the sender immediately by > replying to this message and then delete it from your system. Any > review, dissemination, distribution, or reproduction of this > message by unintended recipients is strictly prohibited and may be > subject to legal restriction. > > > Thank you for your cooperation. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > --------------------------------- > Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & > more on new and used cars. > > ------------------------------ > > Message: 10 > Date: Mon, 23 Jan 2006 15:59:24 -0700 > From: Gayle Callis > Subject: [Histonet] Re: How to get good cross sections of the mouse > small intestine > To: "Yu, Jian" , Histonet@lists.utsouthwestern.edu > Message-ID: > <6.0.0.22.1.20060123154522.01b2cec0@gemini.msu.montana.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Jian, > > You need to rinse the feces out of the lumen. Remove the small > intestine > INTACT but carefully by lifting it out of mouse. You can detach the > mesentary as you do this but do not rip the sample out, you do NOT > want to > tear holes in the intestine. Fill a 50 ml syringe with PBS, and > use a > dulled 18 guage needle - insert into one end of the intestine > (easier from > the ilium side) and inject the PBS. After rinsing out feces, use a > syringe > filled with neutral buffered formalin, and inject that so the lumen > distends. This will fix the villi much faster and get rid of > digestive > enzymes and left over fecal matter. > > You can clamp off the ends of the intestine or have sutures ready > to tie > off ends but you need to do this as the fixative is flowing out the > opposite end of the intestine - overdistending the gut may not be a > good > idea, you need just enough to fix but not create a huge sausage like > structure. Someone helping is nice, as they can tie off one end > while the > needle is removed to tie off the other end or use mosquito > hemostats and > clamp off. The idea is to fix the inside of intestine quickly to > preserve > villi. The gut can be stretched out in distended state or you can > drop it > into fixative then cut cross sections from each individual area of > interest, process and embed on end. WE do this but NOT for cross > sections, > but mid sagittal sections however the intestine is well fixed, and > with > clean results. > > After fixation, you can store in 70% alcohol until processing. > Just make > sure you have total fixation or the 70% will finish it off for > you. We are > careful to not over process these tissues, they are small and > delicate, and > use a shorter processing schedule than for larger tissues. > > At 02:11 PM 1/23/2006, you wrote: >> Dear all, >> >> I am a new histoneter. I am just starting to do some mouse small >> intestine histopathology and wondering whether anyone can provide a >> method (protocol) for how to fix and embed mouse small intestine to >> generate good cross (paraffin) sections with as many intact villi and >> crypts possible for H&E and IHC Particularly, I need to evaluate at >> least 5-10 different area of the small intestine to score the >> regeneration of crypts and the extend of apoptosis at various time >> point >> after irradiation. >> >> I have used neutral buffered formalin as fixative and manually bundle >> four pieces of 0.5 cm-long tubes together before embedding at a >> clinical >> pathology lab. The sections looked OK, but many appear to have a >> lot of >> floating villi in the lumen and limited number of crypts extending >> into >> to the villi connected to them. Since we can not always get our >> tissues processed after O/N fixation, we sometimes keep the >> tissues in >> NBF for several weeks. Is this too long, or should we transfer >> them to >> 70% ethanol? > > Gayle Callis > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 > 406 994-4303 (FAX) > > > > > > ------------------------------ > > Message: 11 > Date: Mon, 23 Jan 2006 17:22:21 -0600 > From: "anita dudley" > Subject: RE: [Histonet] H.pylori immunos on gastric biopsies > To: RCazares@schosp.org, histonet@pathology.swmed.edu > Message-ID: > Content-Type: text/plain; format=flowed > > ruth, we do immuno on all of our gastric bx's. we do them every > day, the > tech that comes in early cuts them and puts them in the 58 oven. > the drs > like them coming to them with the case. I was in a lecture at nat. > meeting > and the person said that the immuno stain was one of the best ways to > demonstrate them. I could probably find the lecture for you in my > national > notebooks, if you like. > anita dudley > providence hosp > mobile alabama > > >> From: "Cazares, Ruth" >> To: >> Subject: [Histonet] H.pylori immunos on gastric biopsies >> Date: Mon, 23 Jan 2006 15:34:27 -0600 >> >> Hello Histonetters, >> >> >> >> I was wondering if anyone out there is doing immuno staining >> routinely >> on all gastric bx's? We are considering this in my lab, but I >> have my >> reservations. If some of you do, how many a week? And those who >> don't, >> why not? I will be doing a literature search on this topic, but if >> anyone knows of a specific article I sure would appreciate it if you >> could share it with me. >> >> >> >> Thank you, >> >> >> >> Ruth Cazares >> >> >> >> >> >> *** Confidentiality Statement *** >> This e-mail is intended only for the use of the individual or >> entity to >> which it is addressed and may contain information that is >> privileged and >> confidential. If the reader of this message is not the intended >> recipient, >> please notify the sender immediately by replying to this message >> and then >> delete it from your system. Any review, dissemination, >> distribution, or >> reproduction of this message by unintended recipients is strictly >> prohibited and may be subject to legal restriction. >> >> >> Thank you for your cooperation. >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > ------------------------------ > > Message: 12 > Date: Mon, 23 Jan 2006 18:44:44 -0500 > From: "Monfils, Paul" > Subject: RE: [Histonet] How to get good cross sections of the mouse > small intestine > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > > <09C945920A6B654199F7A58A1D7D1FDE01717654@lsexch.lsmaster.lifespan.org > > > > Content-Type: text/plain; charset="iso-8859-1" > > In addition to Gayle's suggestions, you might want to physically > hold the intestine straight while the formalin fixes and hardens > it. After > injecting formalin as Gayle said, I pin one end of it in a wax-lined > dissection pan, then stretch it gently, just enough to straighten > it, and > pin the other end to keep it straight, then cover it with formalin. > People > sometimes send me intestine specimens which they have simply > removed from > the animal and dropped into formalin, resulting in a hardened > gordian knot, > all parts of which are curved to some degree, so that pieces cut > from the > intestine are shaped like parentheses. It's difficult to get a good > perpendicular section of a curved object. If you don't have a > dissecting > pan you can easily make one; or you can just pin the specimen onto > a strip > of heavy corrugated cardbord or soft wood and drop it specimen side > down > into a dish of formalin. > > > > ------------------------------ > > Message: 13 > Date: Mon, 23 Jan 2006 17:40:50 -0700 > From: IGOR NASONKIN > Subject: Re: [Histonet] Human specific markers > To: Jim Manavis > Cc: Histonet > Message-ID: <3a4109cfaf1.43d51522@jhmimail.jhmi.edu> > Content-Type: text/plain; charset=us-ascii > > > Jim, > we use HNu (Human nuclei AB) from Chemicon; very robust, stains > well and very specific. > Staining is nuclear. > > Dr. Igor O. Nasonkin Ph.D. > Postdoctoral Fellow > Pathology/Neuropathology > Johns Hopkins University > Ross Research Bldg Rm 563 > 720 Rutland Ave > Baltimore MD 21205 > tel: 617-388-4104 > Igor.Nasonkin@jhmi.edu > > ----- Original Message ----- > From: Jim Manavis > Date: Monday, January 23, 2006 2:53 pm > Subject: [Histonet] Human specific markers > >> Can anyone suggest a human specific marker for use in >> Immnohistochemistry to >> distinguish between human and mouse cells >> >> Jim >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > ------------------------------ > > Message: 14 > Date: Mon, 23 Jan 2006 17:34:45 -0800 (PST) > From: Glenn Krasinski > Subject: [Histonet] Source for "Citrusolve"... > To: histonet@lists.utsouthwestern.edu > Message-ID: <20060124013445.73779.qmail@web37107.mail.mud.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > I am looking for a source for "citrusolve" for use instead of > xylene in the deparaffinization of slides. Does it go by any other > name? > > Many thanks. > > Glenn M. Krasinski > San Diego, CA > > > > > --------------------------------- > Yahoo! Photos ? Showcase holiday pictures in hardcover > Photo Books. You design it and we?ll bind it! > > ------------------------------ > > Message: 15 > Date: Mon, 23 Jan 2006 20:52:02 -0500 > From: "Bartlett, Jeanine" > Subject: RE: [Histonet] Source for "Citrusolve"... > To: "Glenn Krasinski" , > > Message-ID: > > Content-Type: text/plain; charset="UTF-8" > > It's Citrisolve and Amity International carries it....probably some > others do as well. > > http://ascenaa2.miniserver.com/~amity/products/citrisolve.htm > > Jeanine Bartlett > CDC, Atlanta > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Glenn > Krasinski > Sent: Mon 1/23/2006 8:34 PM > To: histonet@lists.utsouthwestern.edu > Cc: > Subject: [Histonet] Source for "Citrusolve"... > > > > I am looking for a source for "citrusolve" for use instead of > xylene in the deparaffinization of slides. Does it go by any other > name? > > Many thanks. > > Glenn M. Krasinski > San Diego, CA > > > > > --------------------------------- > Yahoo! Photos ??? Showcase holiday pictures in hardcover > Photo Books. You design it and we???ll bind it! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 16 > Date: Tue, 24 Jan 2006 10:07:50 +0100 (CET) > From: eva-moeller@arcor.de > Subject: [Histonet] Aw: BrdU Staining HCl pretreat > To: histonet@lists.utsouthwestern.edu > Message-ID: > <4566989.1138093670423.JavaMail.ngmail@webmail-07.arcor-online.net> > Content-Type: text/plain; charset=ISO-8859-1 > > > > hello again, > I forgot to mention that I cook the slices in citrate buffer for 5 > min (800 Watts in microwave) before applying the antibody. > I am now trying to do the HCl method instead. > I will report my results. > Thanks for helping me. > Eva > > Eva Moeller > Ruhr-Universitaet Bochum > Germany > > Machen Sie aus 14 Cent spielend bis zu 100 Euro! > Die neue Gaming-Area von Arcor - ?ber 50 Onlinespiele im Angebot. > http://www.arcor.de/rd/emf-gaming-1 > > > > ------------------------------ > > Message: 17 > Date: Tue, 24 Jan 2006 07:53:19 -0600 > From: "GUTIERREZ, JUAN" > Subject: RE: [Histonet] H.pylori immunos on gastric biopsies > To: "Cazares, Ruth" , > > Message-ID: > > Content-Type: text/plain; charset="US-ASCII" > > We do immunos on all gastric biopsies. We had some cases that looked > negative by giemsa stain, yet the clinical hx suggested otherwise. > When > we ran the immunos on these so called negatives, we were shocked to > see > the amount of organisms we had missed. Do you want to take that > chance? > I always ask myself: what if this was my biopsy? Something to think > about. > > Juan C. Gutierrez, HT(ASCP) > Histology Laboratory Supervisor > (210)704-2533 > > My opinions are my own and do not reflect those of my employer. Long > live free speech! > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Cazares, > Ruth > Sent: Monday, January 23, 2006 3:34 PM > To: histonet@pathology.swmed.edu > Subject: [Histonet] H.pylori immunos on gastric biopsies > > Hello Histonetters, > > > > I was wondering if anyone out there is doing immuno staining routinely > on all gastric bx's? We are considering this in my lab, but I have my > reservations. If some of you do, how many a week? And those who > don't, > why not? I will be doing a literature search on this topic, but if > anyone knows of a specific article I sure would appreciate it if you > could share it with me. > > > > Thank you, > > > > Ruth Cazares > > > > > > *** Confidentiality Statement *** > This e-mail is intended only for the use of the individual or > entity to > which it is addressed and may contain information that is > privileged and > confidential. If the reader of this message is not the intended > recipient, please notify the sender immediately by replying to this > message and then delete it from your system. Any review, > dissemination, > distribution, or reproduction of this message by unintended recipients > is strictly prohibited and may be subject to legal restriction. > > > Thank you for your cooperation. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 18 > Date: Tue, 24 Jan 2006 09:24:33 -0500 > From: bsylinda@aol.com > Subject: [Histonet] CAP Question about Microwave > To: histonet@lists.utsouthwestern.edu > Message-ID: <8C7EF090810EA74-968-20369@mblk-d37.sysops.aol.com> > Content-Type: text/plain; charset="us-ascii" > > Hello Histoland, > I am writing to ask if all labs are using lab approved microwaves > and have them vented to an outside source. Are there still labs > using microwaves from local retailers. If not how are you > addressing the new CAp guidelines on checklist. > > Thanks in advance, > Sylinda Battle, HT (ASCP) > > > ------------------------------ > > Message: 19 > Date: Tue, 24 Jan 2006 09:39:44 -0500 > From: Tim Wheelock > Subject: [Histonet] UN-SUBSCRIBE PLEASE > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: <43D63C30.1060105@mclean.harvard.edu> > Content-Type: text/plain; charset=ISO-8859-1; format=flowed > > I will be going on vacation. > Please unsubscribe me. > > Thanks > > > Any information, including protected health information (PHI), > transmitted > in this email is intended only for the person or entity to which > it is > addressed and may contain information that is privileged, > confidential and or > exempt from disclosure under applicable Federal or State law. Any > review, > retransmission, dissemination or other use of or taking of any > action in > reliance upon, protected health information (PHI) by persons or > entities other > than the intended recipient is prohibited. If you received this > email in error, > please contact the sender and delete the material from any computer. > > > > ------------------------------ > > Message: 20 > Date: Tue, 24 Jan 2006 10:05:28 -0500 > From: "Slyter, Rodney" > Subject: [Histonet] job opening > To: > Cc: "Hedgepeth, Alice" , "Goodrich, Jesse" > > Message-ID: > > <95B1FC82E0F69D4FB1D2C04AAAF6CD721C7B1C@rivhsexchange.rhs.rivhs.local> > Content-Type: text/plain; charset="iso-8859-1" > > There is an immediate opening for a histotech at riverside regional > medical center in Newport news Virginia. This is a full-time > morning position for a certified or eligible histotech. We provide > a wide variety of tests including a wide battery of immunos, iSH > and FISH. We do approx. 16,000 specimens a year both large > resections and biopsies. There is no weekend or call coverage. > There are two other full time techs, two full time lab aids, and a > certified PA who is also a certified histotech. You can email me > directly or visit the Riverside regional medical center website for > more information. > > Rod Slyter, HTL(ASCP), PA(AAPA) > Anatomical Pathology Manager > Riverside Regional Medical Center > > > > 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. > > ------------------------------ > > Message: 21 > Date: Tue, 24 Jan 2006 10:06:52 -0500 > From: kjsavage@buffalo.edu > Subject: [Histonet] (no subject) > To: histonet@lists.utsouthwestern.edu > Message-ID: <1138115212.43d6428c1aea3@mail3.buffalo.edu> > Content-Type: text/plain > > Greetings all, > > I am having a difficult time trying to make methacarn fixative. I > have > searched the list archives and found a posting in May 2005 from > someone > that was having the same problem I am facing: a white precipitate > forming. In one of the responses to his posting it was suggested to > use glass only (pipets, vials etc...). I have done that, and still > the > precipitate forms. I have also used all new reagents (chloroform, > methanol, acetic acid) and still the precipitate forms. Does anyone > have some more suggestions as to what is going wrong? Thanks, Kathy > > Kathy Savage, PhD > Dept. of Exercise and Nutrition Sciences > SUNY-University at Buffalo > > > > ------------------------------ > > Message: 22 > Date: Tue, 24 Jan 2006 09:29:19 -0600 > From: Bill Blank > Subject: RE: [Histonet] H.pylori immunos on gastric biopsies > To: > Message-ID: > Content-Type: text/plain; charset="us-ascii" ; format="flowed" > > IMO, we take this detecting H. pylori thing too seriously. A rational > physician treats the patient and not any lab test. If I had symptoms > of H. pylori, if my stomach looked inflamed, I would want to be > treated for H. pylori irregardless of their presence on a biopsy. > (Well, maybe as there is some evidence that treating antral H. pylori > may increase the risk of GE junction adenocarcinoma) > > I consider immunos on gastric biopsies to be overkill and a waste of > health care dollars. > > BIll > > At 7:53 AM -0600 1/24/06, GUTIERREZ, JUAN wrote: >> We do immunos on all gastric biopsies. We had some cases that looked >> negative by giemsa stain, yet the clinical hx suggested >> otherwise. When >> we ran the immunos on these so called negatives, we were shocked >> to see >> the amount of organisms we had missed. Do you want to take that >> chance? >> I always ask myself: what if this was my biopsy? Something to think >> about. > > -- > _____________________________ > Bill Blank > http://kernunnos.com (Celtic studies and numismatics) > OBOD's Message board: http://www.druidry.org/board > > > > > > > ------------------------------ > > Message: 23 > Date: Tue, 24 Jan 2006 08:50:35 -0700 > From: Gayle Callis > Subject: RE: [Histonet] How to get good cross sections of the mouse > small intestine > To: "Monfils, Paul" , > Histonet@lists.utsouthwestern.edu > Message-ID: > <6.0.0.22.1.20060124083902.01b439c8@gemini.msu.montana.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Paul, > > Excellent suggestion - for pinning down the samples, I suggest using > disposable hypodermic needles - the latter do NOT rust or corrode in > aqueous based NBF - needles are stainless steel. Tiny needles (24 > or less > gauge) for this purpose and not tear the ends of the intestine to > bits > with larger gauge. > > The Gordian knot description was so true and our loops of intestine > always > were filled with mouse feces!!! Once fixed, couldn't removed - > microtomy > was no fun and sections looked terrible. > > You can also use cork strips (purchased from hardware store, very > cheap) > then toss them out later. We would stand these on end in a deep > bucket > with lid - tagged carefully. And a cheap pan with a lid is the > cake pan > sizes or larger, with lids - Rubbermaid or some other plastic. > Very nice > to reach into a pan and get the samples - loved this idea. > > > > At 04:44 PM 1/23/2006, you wrote: >> In addition to Gayle's suggestions, you might want to >> physically >> hold the intestine straight while the formalin fixes and hardens >> it. After >> injecting formalin as Gayle said, I pin one end of it in a wax-lined >> dissection pan, then stretch it gently, just enough to straighten >> it, and >> pin the other end to keep it straight, then cover it with >> formalin. People >> sometimes send me intestine specimens which they have simply >> removed from >> the animal and dropped into formalin, resulting in a hardened >> gordian knot, >> all parts of which are curved to some degree, so that pieces cut >> from the >> intestine are shaped like parentheses. It's difficult to get a good >> perpendicular section of a curved object. If you don't have a >> dissecting >> pan you can easily make one; or you can just pin the specimen onto >> a strip >> of heavy corrugated cardbord or soft wood and drop it specimen >> side down >> into a dish of formalin. >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis HTL, HT, MT(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University > Bozeman MT 59717 > > > > > ------------------------------ > > Message: 24 > Date: Tue, 24 Jan 2006 10:57:06 -0500 > From: bsylinda@aol.com > Subject: Re: [Histonet] CAP Question about Microwave > To: bsylinda@aol.com, histonet@lists.utsouthwestern.edu > Message-ID: <8C7EF15F587F78E-A58-150A6@FWM-D06.sysops.aol.com> > Content-Type: text/plain; charset="us-ascii" > > An additionan to comment. This microwave is only used for special > stains no tissuse processing. > Thanks again, > sylinda > > -----Original Message----- > From: bsylinda@aol.com > To: histonet@lists.utsouthwestern.edu > Sent: Tue, 24 Jan 2006 09:24:33 -0500 > Subject: [Histonet] CAP Question about Microwave > > > Hello Histoland, > I am writing to ask if all labs are using lab approved microwaves > and have them > vented to an outside source. Are there still labs using microwaves > from local > retailers. If not how are you addressing the new CAp guidelines on > checklist. > > Thanks in advance, > Sylinda Battle, HT (ASCP) > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > ------------------------------ > > Message: 25 > Date: Tue, 24 Jan 2006 10:17:45 -0600 > From: "Smith, Jeffery D. \(HSC\)" > Subject: RE: [Histonet] H.pylori immunos on gastric biopsies > To: "Bill Blank" , > > Message-ID: > > Content-Type: text/plain; charset="iso-8859-1" > > I have to somewhat agree. Immunos for H. pylori seems to be > overkill. We routinely stain for H. pylori using a Giemsa on all > GI biopsies. > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bill > Blank > Sent: Tue 1/24/2006 9:29 AM > To: histonet@pathology.swmed.edu > Subject: RE: [Histonet] H.pylori immunos on gastric biopsies > > > > IMO, we take this detecting H. pylori thing too seriously. A rational > physician treats the patient and not any lab test. If I had symptoms > of H. pylori, if my stomach looked inflamed, I would want to be > treated for H. pylori irregardless of their presence on a biopsy. > (Well, maybe as there is some evidence that treating antral H. pylori > may increase the risk of GE junction adenocarcinoma) > > I consider immunos on gastric biopsies to be overkill and a waste of > health care dollars. > > BIll > > At 7:53 AM -0600 1/24/06, GUTIERREZ, JUAN wrote: >> We do immunos on all gastric biopsies. We had some cases that looked >> negative by giemsa stain, yet the clinical hx suggested >> otherwise. When >> we ran the immunos on these so called negatives, we were shocked >> to see >> the amount of organisms we had missed. Do you want to take that >> chance? >> I always ask myself: what if this was my biopsy? Something to think >> about. > > -- > _____________________________ > Bill Blank > http://kernunnos.com (Celtic studies and numismatics) > OBOD's Message board: http://www.druidry.org/board > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 26 > Date: Tue, 24 Jan 2006 11:27:14 -0500 > From: "Sue Barnes" > Subject: [Histonet] Health Testing when using xylene > To: > Message-ID: > Content-Type: text/plain; charset="Windows-1252" > > > Does anyone in the histology world have health testing for techs > who use xylene? > What testing is done? > Is there printed regulations for suggested health testing? > Any help on this would be appreciated. > > Thanks > Sue Barnes > East Liverpool City Hospital > East Liverpool, Ohio > > > ------------------------------ > > Message: 27 > Date: Tue, 24 Jan 2006 11:32:29 -0500 > From: Phil McArdle > Subject: Re: [Histonet] CAP Question about Microwave > To: histonet@lists.utsouthwestern.edu > Message-ID: <43D6569D.5010309@ebsciences.com> > Content-Type: text/plain; charset=ISO-8859-1; format=flowed > > Hello: > > If you want some "vendor" input, please feel free to contact me. > > Best regards, > > Phil McArdle > > -- > Phil McArdle > Microwave Product Manager > Energy Beam Sciences, Inc. > Tel: 800.992.9037 x 341 > Fax: 860.653.0422 > PMcardle@ebsciences.com > www.ebsciences.com > "ADDING BRILLIANCE TO YOUR VISION" > > > > > ------------------------------ > > Message: 28 > Date: Tue, 24 Jan 2006 08:48:28 -0800 (PST) > From: Rene J Buesa > Subject: Re: [Histonet] Health Testing when using xylene > To: Sue Barnes , histonet@lists.utsouthwestern.edu > Message-ID: <20060124164828.11569.qmail@web61214.mail.yahoo.com> > Content-Type: text/plain; charset=iso-8859-1 > > Sue: > I used to test for formaldehyde, glutaraldehyde, xylene and the > air flow level of the fumes hoods. > The fumes hoods had simple air gauges tha were checked daily and > the results written in a log in the hood door. > For the xylene (as well as the others) we used pasive monitoring > badges worn during 8 hours (to determine Time Weighed Average or > TWA) or during 15 minutes (to determine Short Term Exposure Level > or STEL). > Some of the badges were developed in our lab and others were sent > to an outside laboratory for confirmation of results. > After 8 years of cummulative data with values below OSHA levels, > and because this was a very expensive program, we contracted an > outside company that did the monitoring only in the areas and we > stopped doing the personal monitoring. > Hope this will help you!. > Ren? J. > > Sue Barnes wrote: > > Does anyone in the histology world have health testing for techs > who use xylene? > What testing is done? > Is there printed regulations for suggested health testing? > Any help on this would be appreciated. > > Thanks > Sue Barnes > East Liverpool City Hospital > East Liverpool, Ohio > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > --------------------------------- > Yahoo! Photos > Ring in the New Year with Photo Calendars. Add photos, events, > holidays, whatever. > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 26, Issue 27 > **************************************** From DKAEK <@t> coloplast.com Wed Jan 25 02:34:19 2006 From: DKAEK <@t> coloplast.com (Annette Ekblond) Date: Wed Jan 25 02:34:33 2006 Subject: [Histonet] monocyte activation Message-ID: Dear Histonet subscribers Looking at in vitro biocompatibility which histological or rather immunohistochemical markers of monocyte activation or differentiation would you suggest as the most predictive of in vivo performance? Annette Ekblond Denmark From ali.moussavi <@t> molbio.gu.se Wed Jan 25 02:31:57 2006 From: ali.moussavi <@t> molbio.gu.se (Ali Moussavi Nik) Date: Wed Jan 25 02:34:38 2006 Subject: [Histonet] Qu:Assesment of embryonic BBB defect In-Reply-To: <4obt61$57lh2@ironportin-lyk-1.it.gu.se> Message-ID: <000001c62189$ce85f700$274ef182@alis> Dear All Hi Dose any one knows how to asses the embryonic Blood Brain Barrier defect. I would like to see whether or not the Blood Brain Barrier of 18.5 mouse embryo of a nock out strain is leaky. I will extremely thankful if someone gives me a practical protocol. ---------------------------------------------------------------------------- ------ Seyed Ali Moussavi Nik D.V.M , PhD Student Department of Cell and Molecular Biology, Gothenburg University Box 462 S-405 30 G?teborg Sweden Phone: +46(031) 7733895 Fax: +46(031) 7733801 www.molbio.gu.se From kemlo.rogerson <@t> waht.swest.nhs.uk Wed Jan 25 02:59:04 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Wed Jan 25 02:59:28 2006 Subject: [Histonet] Zinc Formalin Message-ID: If you leave the tissue in too long it will become hard and, if my memory serves me, you may have problems with overstaining with haematoxylin. My advice, if you can't get it back, is to take the tissue out of the fixative ASAP and put into regular fixative. Kemlo Rogerson West Super Mare Pathology Service Manager (recovering Cell Path Manager) -----Original Message----- From: Laurie Colbert [mailto:laurie.colbert@huntingtonhospital.com] Sent: Tuesday, January 24, 2006 11:13 PM To: Histonet (E-mail) Subject: [Histonet] Zinc Formalin Our formalin supplier mistakenly shipped us pre-filled containers containing zinc formalin instead of 10% NBF. Before I noticed this, three cases had been distributed throughout the hospital (most likely to Surgery). So, how is this going to affect my tissue? Is there any post-treatment (such as removing mercury pigment on B-5 fixed tissue) that I need to perform? Thanks in advance for any info you can provide. Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Wed Jan 25 03:02:07 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Wed Jan 25 03:02:25 2006 Subject: [Histonet] H.pylori immunos on gastric biopsies Message-ID: Isn't there a breath test too? Kemlo Rogerson Weston Super Mare Cell Path Manager -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, January 24, 2006 5:09 PM To: Smith, Jeffery D. (HSC); Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies H. pylori is a pathogen. It does not sit in the stomach behaving itself. Its presence is an indication for treatment, but its absence is *not* an indication for treatment directed against it, irrespective of how inflamed the stomach looks. The best histologic determinant of H. pylori is an immunostain. However, in most cases, this is all rather academic, as a quick stool test is available, directed against H. pylori antigen, so in most cases, we are really trying to determine something better done by other means. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Smith, Jeffery D. (HSC) [mailto:Jeffery-Smith@ouhsc.edu] Sent: 24 January 2006 16:18 To: Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies I have to somewhat agree. Immunos for H. pylori seems to be overkill. We routinely stain for H. pylori using a Giemsa on all GI biopsies. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bill Blank Sent: Tue 1/24/2006 9:29 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies IMO, we take this detecting H. pylori thing too seriously. A rational physician treats the patient and not any lab test. If I had symptoms of H. pylori, if my stomach looked inflamed, I would want to be treated for H. pylori irregardless of their presence on a biopsy. (Well, maybe as there is some evidence that treating antral H. pylori may increase the risk of GE junction adenocarcinoma) I consider immunos on gastric biopsies to be overkill and a waste of health care dollars. BIll At 7:53 AM -0600 1/24/06, GUTIERREZ, JUAN wrote: >We do immunos on all gastric biopsies. We had some cases that looked >negative by giemsa stain, yet the clinical hx suggested otherwise. When >we ran the immunos on these so called negatives, we were shocked to see >the amount of organisms we had missed. Do you want to take that chance? >I always ask myself: what if this was my biopsy? Something to think >about. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@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 peter_bannister <@t> hotmail.co.uk Wed Jan 25 03:42:56 2006 From: peter_bannister <@t> hotmail.co.uk (Peter Bannister) Date: Wed Jan 25 03:43:05 2006 Subject: [Histonet] PLEASE HELP Replacement imaging monitor for Biorad MRC 600 Confocal microscope Message-ID: Hello Histonetters, Does anyone know the make and model of a suitable replacement for the broken imaging monitor I have attached to my MRC 600 confocal microscope? The problem is that the system is ancient (but all we've got for the moment), Biorad no longer exist, and the system will only work with a slow scan monitor that must be connected via a standard BNC RGB video cable. I have found some Sony Trinitron CCTV monitors on the web but have no way of knowing for sure if they will work before I make the purchase. Do any of you kind Histonetters use this Biorad system? What model is your imaging monitor? Any help will be very much appreciated. Peter Bannister _________________________________________________________________ Are you using the latest version of MSN Messenger? Download MSN Messenger 7.5 today! http://messenger.msn.co.uk From Mary.Judd <@t> suht.swest.nhs.uk Wed Jan 25 04:32:42 2006 From: Mary.Judd <@t> suht.swest.nhs.uk (Mary Judd) Date: Wed Jan 25 04:33:02 2006 Subject: [Histonet] TNFa, IL6, VEGF, Oxytocin antibodies Message-ID: TNFa, IL6, VEGF, Oxytocin antibodies Has anyone used any of these antibodies for IHC on colon. I would particularily appreciate advice on antigen retrieval. These antibodies are new to us, we have been asked to stain them for somebody's research project, and we are not even sure about localization. In a word Help! Mary Judd Mary Judd Section Head Immunohistochemistry Cellular Pathology Level E, Mailpoint 2 Southampton University Hospitals Trust Tel. 02380 795144 Fax. 02380 796869 D I S C L A I M E R 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. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Trust unless explicitly stated otherwise. If you have received this e-mail in error please delete the e-mail and contact the Southampton University Hospitals NHS Trust Helpdesk on:- 023 80796000 The information contained in this e-mail may be subject to public disclosure under the Freedom of Information Act 2000. Unless the Information is legally exempt from disclosure, the confidentiality of this e-mail and your reply cannot be guaranteed. This footnote also confirms that this email message has been swept by MailMarshal for the presence of computer viruses. Please visit our website at http://www.suht.nhs.uk From Terry.Marshall <@t> rothgen.nhs.uk Wed Jan 25 05:54:24 2006 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Wed Jan 25 05:53:07 2006 Subject: [Histonet] H.pylori immunos on gastric biopsies Message-ID: Yes, but a turd is easier to bottle. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:kemlo.rogerson@waht.swest.nhs.uk] Sent: 25 January 2006 09:02 To: Marshall Terry Dr, Consultant Histopathologist; Smith, Jeffery D. (HSC); Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies Isn't there a breath test too? Kemlo Rogerson Weston Super Mare Cell Path Manager -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, January 24, 2006 5:09 PM To: Smith, Jeffery D. (HSC); Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies H. pylori is a pathogen. It does not sit in the stomach behaving itself. Its presence is an indication for treatment, but its absence is *not* an indication for treatment directed against it, irrespective of how inflamed the stomach looks. The best histologic determinant of H. pylori is an immunostain. However, in most cases, this is all rather academic, as a quick stool test is available, directed against H. pylori antigen, so in most cases, we are really trying to determine something better done by other means. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Smith, Jeffery D. (HSC) [mailto:Jeffery-Smith@ouhsc.edu] Sent: 24 January 2006 16:18 To: Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies I have to somewhat agree. Immunos for H. pylori seems to be overkill. We routinely stain for H. pylori using a Giemsa on all GI biopsies. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bill Blank Sent: Tue 1/24/2006 9:29 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies IMO, we take this detecting H. pylori thing too seriously. A rational physician treats the patient and not any lab test. If I had symptoms of H. pylori, if my stomach looked inflamed, I would want to be treated for H. pylori irregardless of their presence on a biopsy. (Well, maybe as there is some evidence that treating antral H. pylori may increase the risk of GE junction adenocarcinoma) I consider immunos on gastric biopsies to be overkill and a waste of health care dollars. BIll At 7:53 AM -0600 1/24/06, GUTIERREZ, JUAN wrote: >We do immunos on all gastric biopsies. We had some cases that looked >negative by giemsa stain, yet the clinical hx suggested otherwise. When >we ran the immunos on these so called negatives, we were shocked to see >the amount of organisms we had missed. Do you want to take that chance? >I always ask myself: what if this was my biopsy? Something to think >about. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Wed Jan 25 06:57:18 2006 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Wed Jan 25 06:57:52 2006 Subject: [Histonet] Re: Need for antigen retrieval question Message-ID: Glenn, What an interesting question! I would imagine you would not need to antigen retrieve the sample (i.e. mouse on mouse secondary antibody issue). However, you might do a side-by-side comparison to see if there is any difference in signal. Do let us know how you fare! Teri Johnson, HT(ASCP)QIHC Stowers Institute for Medical Research Kansas City, MO 64110 From TMcNemar <@t> lmhealth.org Wed Jan 25 07:09:56 2006 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Wed Jan 25 07:15:24 2006 Subject: [Histonet] H.pylori immunos on gastric biopsies Message-ID: <6CD94D97ED7D924BA5C2B588FA952821396835@nt_exchange.lmhealth.org> We switched to IHC for H. Pylori years ago. Nobody was happy with the stains we used before. IHC makes it so easy with almost zero repeats (assuming your control has bugs). We do them everyday and probaly 25 -35 per week. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Cazares, Ruth [mailto:RCazares@schosp.org] Sent: Monday, January 23, 2006 4:34 PM To: histonet@pathology.swmed.edu Subject: [Histonet] H.pylori immunos on gastric biopsies Hello Histonetters, I was wondering if anyone out there is doing immuno staining routinely on all gastric bx's? We are considering this in my lab, but I have my reservations. If some of you do, how many a week? And those who don't, why not? I will be doing a literature search on this topic, but if anyone knows of a specific article I sure would appreciate it if you could share it with me. Thank you, Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lcastillos <@t> icoria.com Wed Jan 25 07:17:53 2006 From: lcastillos <@t> icoria.com (Castillos, Luminita) Date: Wed Jan 25 07:18:07 2006 Subject: [Histonet] Hematoxylin-eosin-saffron staining Message-ID: Hi, I am looking to find the protocol for the hematoxylin-eosin-saffron method and I have difficulties. I would like to ask if anybody can share with me this protocol?. I appreciated so much. Also, does somebody knows a staining for macrophages from the lung or synovial tissue , using only histochemistry staining without immunostaining?. Thank you again. Sincerely, Luminita Luminita Castillos, Ph.D. Research Scientist Icoria, Inc., A Clinical Data Inc. company 108 T.W. Alexander Drive P.O. Box 14528 RTP, NC 27709-4528 Phone: 919-425-2967 Cell: 919-302-0485 www.icoria.com lcastillos@icoria.com Effective Dec. 21, 2005, Icoria became a Clinical Data, Inc. company, the worldwide leader in the molecular diagnostic field. The Pharmacogenomics and the Molecular Services Division of Clinical Data, Inc. (now with expanded genomics offerings from Icoria) specializes in the development and commercialization of molecular biology and pharmacogenomics solutions to biotechnology and pharmaceutical companies, academia and government institutions. From rjbuesa <@t> yahoo.com Wed Jan 25 07:23:33 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Jan 25 07:23:42 2006 Subject: [Histonet] Need for antigen retrieval question... In-Reply-To: <20060124230503.59072.qmail@web37111.mail.mud.yahoo.com> Message-ID: <20060125132333.72250.qmail@web61223.mail.yahoo.com> Glenn: The tissues fixed in formalin undergo protein crosslinkage and need antigen retrieval (HIER). In a regular IHC procedure you apply the primary antibody trying to reveal the antigen epitope that has been crosslinked by using HIER. In your experiment the epitope should heve been detected in vivo but now you are trying to detect the antibody bonded to the antigen epitope but both of them (antigen and antibody) are proteins that have been crosslinked by the formalin, and you have to "unravel" them during HIER. Ren? J. Glenn Krasinski wrote: My boss will be injecting mice burdened with solid tumors with antibodies specific to potential biomarkers. The tumors/tissues from the sacrificed animals will be fixed in formalin and embedded in paraffin blocks. Sections will then be stained with secondary antibodies to the specific biomarker antibodies. My question is whether or not I will need to go through an antigen retrieval process. My apologies for the complicated question. Glenn M. Krasinski San Diego, CA 92121 --------------------------------- Yahoo! Photos Got holiday prints? See all the ways to get quality prints in your hands ASAP. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with your Yahoo! Mail. From rjbuesa <@t> yahoo.com Wed Jan 25 08:07:49 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Jan 25 08:07:58 2006 Subject: [Histonet] Hematoxylin-eosin-saffron staining In-Reply-To: Message-ID: <20060125140749.97774.qmail@web61218.mail.yahoo.com> Hi Luminita: There is a procedure by Langeron (1942) for saffron and eosin B: Sol.A = 1% eosin; Sol.B = dist.water??100mL + saffron??2 g + + 5% aq. tanin??1 mL + 40% formaldehyde?? 1 mL Procedure: hydrated sections stained with hematoxylin ??differentiated ?? nuclei blue ?? sol A for 5 min?? quick rinse in water ?? 70% ethanol for a few seconds ?? wash well in water ?? sol B for 5 min?? BLOT sections ?? flood the sections with absolute ethanol until dehydrated ?? xylene ?? cover slip. For the macrophages I would stain toluidine blue. Hope this will help. Ren?? J. "Castillos, Luminita" wrote: Hi, I am looking to find the protocol for the hematoxylin-eosin-saffron method and I have difficulties. I would like to ask if anybody can share with me this protocol?. I appreciated so much. Also, does somebody knows a staining for macrophages from the lung or synovial tissue , using only histochemistry staining without immunostaining?. Thank you again. Sincerely, Luminita Luminita Castillos, Ph.D. Research Scientist Icoria, Inc., A Clinical Data Inc. company 108 T.W. Alexander Drive P.O. Box 14528 RTP, NC 27709-4528 Phone: 919-425-2967 Cell: 919-302-0485 www.icoria.com lcastillos@icoria.com Effective Dec. 21, 2005, Icoria became a Clinical Data, Inc. company, the worldwide leader in the molecular diagnostic field. The Pharmacogenomics and the Molecular Services Division of Clinical Data, Inc. (now with expanded genomics offerings from Icoria) specializes in the development and commercialization of molecular biology and pharmacogenomics solutions to biotechnology and pharmaceutical companies, academia and government institutions. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. From kmerriam2003 <@t> yahoo.com Wed Jan 25 08:14:06 2006 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Wed Jan 25 08:14:17 2006 Subject: [Histonet] (no subject) In-Reply-To: <1138115212.43d6428c1aea3@mail3.buffalo.edu> Message-ID: <20060125141407.87589.qmail@web50301.mail.yahoo.com> Kathy, I have used methacarn quite a bit in the past and we did occasionally get a precipitate. When this happened, we would just filter the solution and then use it. It should be fine if you filter the solution. Remember that it should be kept in the fridge and has a limited shelf life, maybe 2 weeks or so in the fridge. Good luck, Kim Merriam Novartis Pharm. Cambridge, MA kjsavage@buffalo.edu wrote: Greetings all, I am having a difficult time trying to make methacarn fixative. I have searched the list archives and found a posting in May 2005 from someone that was having the same problem I am facing: a white precipitate forming. In one of the responses to his posting it was suggested to use glass only (pipets, vials etc...). I have done that, and still the precipitate forms. I have also used all new reagents (chloroform, methanol, acetic acid) and still the precipitate forms. Does anyone have some more suggestions as to what is going wrong? Thanks, Kathy Kathy Savage, PhD Dept. of Exercise and Nutrition Sciences SUNY-University at Buffalo _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. From cwscouten <@t> myneurolab.com Wed Jan 25 09:00:49 2006 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Wed Jan 25 09:01:29 2006 Subject: [Histonet] Sliding Microtome operation Message-ID: <5784D843593D874C93E9BADCB87342AB916B43@tpiserver03.Coretech-holdings.com> A customer has asked the following question. Can anybody help? "Can you recommend on what corner I need establish a knife when I'm working with Vibratome 8000, if I work with Super Mega Cassette and using Feather mikrotome blade S35". Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com From Ronnie_Houston <@t> bshsi.com Wed Jan 25 09:05:35 2006 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Wed Jan 25 09:07:22 2006 Subject: [Histonet] Multi-drug resistance antibodies Message-ID: <00FBB1F3374BE24AAB7ACC6FCC7C617F561F01@bsrexms01.bshsir.com> Does anyone have experience with doing IHC for MDR and MRP (multi-drug resistance-associated protein) and if so what clones/suppliers are you using? Any other details greatly appreciated also, including suitable control info. Thanks Ronnie Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 2877972 (804) 2877906 - fax ronnie_houston@bshsi.com ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone, and permanently delete the original e-mail, attachment(s), and any copies. From gcallis <@t> montana.edu Wed Jan 25 09:27:14 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Jan 25 09:27:31 2006 Subject: [Histonet] PLEASE HELP Replacement imaging monitor for Biorad MRC 600 Confocal microscope In-Reply-To: References: Message-ID: <6.0.0.22.1.20060125082431.01b60ce8@gemini.msu.montana.edu> Peter, There is a confocal listserver out of University of Buffalo that is excellent. Just do a search and pick up the website, then subscribe. It is easy to get on and off, plus the people there will probably have some older system extras floating around in their facility. I think you will get lucky there At 02:42 AM 1/25/2006, you wrote: >Hello Histonetters, >Does anyone know the make and model of a suitable replacement for the >broken imaging monitor I have attached to my MRC 600 confocal microscope? >The problem is that the system is ancient (but all we've got for the >moment), Biorad no longer exist, and the system will only work with a slow >scan monitor that must be connected via a standard BNC RGB video cable. I >have found some Sony Trinitron CCTV monitors on the web but have no way of >knowing for sure if they will work before I make the purchase. Do any of >you kind Histonetters use this Biorad system? What model is your imaging >monitor? > >Any help will be very much appreciated. >Peter Bannister > >_________________________________________________________________ >Are you using the latest version of MSN Messenger? Download MSN Messenger >7.5 today! http://messenger.msn.co.uk > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From ree3 <@t> leicester.ac.uk Wed Jan 25 09:29:02 2006 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Wed Jan 25 09:29:12 2006 Subject: [Histonet] RE: GCK-like kinase(GLK) Message-ID: --Anyone have any idea of the distribution of GLK in normal human tissues, unable to find anything in Pubmed. Thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER..U.K..... From mauger <@t> email.chop.edu Wed Jan 25 10:51:20 2006 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Wed Jan 25 10:52:10 2006 Subject: [Histonet] Multi-drug resistance antibodies Message-ID: Ronnie, We use MDR 1+3 from Zymed -clone C219,cat# 18-7244. We use a normal liver control, and high pH (9.5) retrieval. Jo Mauger >>> "Houston, Ronnie" 01/25/06 10:05 AM >>> Does anyone have experience with doing IHC for MDR and MRP (multi-drug resistance-associated protein) and if so what clones/suppliers are you using? Any other details greatly appreciated also, including suitable control info. Thanks Ronnie Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 2877972 (804) 2877906 - fax ronnie_houston@bshsi.com ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone, and permanently delete the original e-mail, attachment(s), and any copies. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mward <@t> wfubmc.edu Wed Jan 25 12:22:22 2006 From: mward <@t> wfubmc.edu (Martha Ward) Date: Wed Jan 25 12:22:31 2006 Subject: [Histonet] Hep Par 1 antibody Message-ID: <61135F0455D33347B5AAE209B903A3041206E7D6@EXCHVS2.medctr.ad.wfubmc.edu> I have approached by our Pathologists about working up the Hep Par 1 antibody and I was wondering if anyone out there is currently doing it, and if so, could you recommend a vendor, conditions, etc. Thanks for much in advance for any help you can give me. Martha Ward, MT (ASCP) QIHC Molecular Diagnostics Lab Wake Forest University Baptist Medical Center From jkiernan <@t> uwo.ca Wed Jan 25 12:26:22 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Wed Jan 25 12:26:33 2006 Subject: [Histonet] Zinc Formalin References: <0BE6ADFAE4E7E04496BF21ABD346628006307C8B@EXCHANGE1.huntingtonhospital.com> Message-ID: <43D7C2CE.54157A1A@uwo.ca> Zinc-formalin mixtures do not contain mercuric chloride, so you will not need to do an iodine and thiosulphate treatment of the sections. Ask your supplier to tell you the composition of his "zinc formalin". Is it an acidic (zinc sulphate or chloride) or a neutral (zinc salicylate) formulation? Most zinc formalin mixtures are acidic, and it may make a difference if the anion is sulphate or chloride. The published literature is clouded by trade secrecy, as are the web sites of those who make the mixtures. The acidic zinc formalin principle is simple: combine a protein coagulant with a cross-linker. All the classical late 19th century fixatives did that, and more. Formalin pigment can be expected if tissues are stored for a long time in an acidic formaldehyde solution. Hope this helps. John Kiernan Anatomy, UWO London, Canada. ______________________________________ Laurie Colbert wrote: > > Our formalin supplier mistakenly shipped us pre-filled containers containing zinc formalin instead of 10% NBF. Before I noticed this, three cases had been distributed throughout the hospital (most likely to Surgery). So, how is this going to affect my tissue? Is there any post-treatment (such as removing mercury pigment on B-5 fixed tissue) that I need to perform? > > Thanks in advance for any info you can provide. > > Laurie Colbert > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed Jan 25 12:29:06 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Wed Jan 25 12:29:02 2006 Subject: [Histonet] (no subject) References: <1138115212.43d6428c1aea3@mail3.buffalo.edu> Message-ID: <43D7C372.2EC99A99@uwo.ca> Dear kjsavage@buffalo.edu Methacarn is easy to make. It contains nothing that could form a precipitate. Your email indicates that you have all the ingredients (chloroform, methanol, acetic acid). This is a non-aqueous coagulant fixative, differing from Carnoy only in having methanol instead of ethanol. Fixatives of this kind contain no water. Cloudiness or a precipitate might happen if an ingredient is wrong or if water gets into the mixture. Are your methanol and acetic acid 95-100%? The advice to avoid plastic was good, because chloroform attacks some plastic ware. John Kiernan Anatomy, UWO London, Canada ______________________________________________________ kjsavage@buffalo.edu wrote: > > Greetings all, > > I am having a difficult time trying to make methacarn fixative. I have > searched the list archives and found a posting in May 2005 from someone > that was having the same problem I am facing: a white precipitate > forming. In one of the responses to his posting it was suggested to > use glass only (pipets, vials etc...). I have done that, and still the > precipitate forms. I have also used all new reagents (chloroform, > methanol, acetic acid) and still the precipitate forms. Does anyone > have some more suggestions as to what is going wrong? Thanks, Kathy > > Kathy Savage, PhD > Dept. of Exercise and Nutrition Sciences > SUNY-University at Buffalo > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Wed Jan 25 12:34:30 2006 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Wed Jan 25 12:34:49 2006 Subject: [Histonet] Hep Par 1 antibody Message-ID: We use DAKO's Hepatocyte antibody, clone OCH1E5. It's very easy to work up on the Ventana stainers. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Martha Ward Sent: Wednesday, January 25, 2006 12:22 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Hep Par 1 antibody I have approached by our Pathologists about working up the Hep Par 1 antibody and I was wondering if anyone out there is currently doing it, and if so, could you recommend a vendor, conditions, etc. Thanks for much in advance for any help you can give me. Martha Ward, MT (ASCP) QIHC Molecular Diagnostics Lab Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Wed Jan 25 12:46:21 2006 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Wed Jan 25 12:46:31 2006 Subject: [Histonet] Hep Par 1 antibody Message-ID: Martha, We use Biocare's Hepatocyte Specific Antibody at 1:50 with a polymer DAB kit on our Ventana automated immunostainers. Our pathologists are happy with it. Linda Sebree, HT(ASCP) University of Wisconsin Hospital & Clinics IHC/ISH Laboratory A4/204-3224 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Martha Ward Sent: Wednesday, January 25, 2006 12:22 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Hep Par 1 antibody I have approached by our Pathologists about working up the Hep Par 1 antibody and I was wondering if anyone out there is currently doing it, and if so, could you recommend a vendor, conditions, etc. Thanks for much in advance for any help you can give me. Martha Ward, MT (ASCP) QIHC Molecular Diagnostics Lab Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed Jan 25 12:48:10 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Wed Jan 25 12:48:11 2006 Subject: [Histonet] Qu:Assesment of embryonic BBB defect References: <000001c62189$ce85f700$274ef182@alis> Message-ID: <43D7C7EA.CFA7A5D5@uwo.ca> Immunostain for a plasma protein such as albumin. It's important to fix the tissue quickly, or plasma will move out of the blood vessels. See Mori et al (1991) Lab. Invest. 64:345-351; Fabian (1992) J. Histochem. Cytochem. 40: 987-991; Loberg & Torvik (1992) Acta path. microbiol. scand. 100: 431-436. I've seen this artifact in human brain, with immunostaining for albumin, ceruloplasmin, orosomucoid and IgG (unpublished). The time of first formation of the BBB is still controversial (far too much literature to mention here) and may depend on the techniques used. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Ali Moussavi Nik wrote: > > Dear All > > Hi > > Dose any one knows how to asses the embryonic Blood Brain Barrier defect. > > I would like to see whether or not the Blood Brain Barrier of 18.5 mouse > embryo of a nock out strain is leaky. > > I will extremely thankful if someone gives me a practical protocol. > > > > > > > > ---------------------------------------------------------------------------- > ------ > > Seyed Ali Moussavi Nik D.V.M , PhD Student > > Department of Cell and Molecular Biology, > > Gothenburg University > > Box 462 > > S-405 30 G?teborg > > Sweden > > Phone: +46(031) 7733895 > > Fax: +46(031) 7733801 > > www.molbio.gu.se > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RCazares <@t> schosp.org Wed Jan 25 13:04:22 2006 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Wed Jan 25 13:04:37 2006 Subject: [Histonet] RE: Helico Immuno Message-ID: <913FAC2B773C19488E26AE6572180FA50458A5CF@exch01.schosp.org> Thanks to all of you who responded to my inquiry on immunos for H. pylori. It really helps to get different opinions and perspectives to help one weigh the pros and cons. Sincerely, Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. From jtrynak <@t> hotmail.com Wed Jan 25 13:30:35 2006 From: jtrynak <@t> hotmail.com (John Rynak) Date: Wed Jan 25 13:30:45 2006 Subject: [Histonet] Wanted : Field Support Specialist - Histopathology, TEXAS Message-ID: Field Support Specialist - Histopathology SciGenium's Client is experiencing tremendous growth within their North American operations. This growth is due to the recent launch of their FDA approved automated histology instrumentation system, and the growing antibody and reagent consumable product line for the histology market. Due to the new launch of these products they are currently looking for Field Support Specialist - Histopathology, for Southwest Region (TX Houston or Dallas ) Based in Texas, this position provides technical applications support for our next generation range of automated histopathology products. This is a very hands on role where your key tasks will include: · Building an in-depth understanding of our new product technologies and their applications in a diagnostic histopathology environment; · Customer liaison by providing scientific and laboratory expertise for in-field installations, training and trouble-shooting; · Customer support Help-line for remote problem solving; · Designing and performing experiments to investigate and solve tough technical applications problems; and · Preparing problem resolution reports and imparting your findings and knowledge to internal and field based Product Engineers and Technical Managers. To be successful in this role you will possess: · Excellent problem solving and analytical skills · The ability to communicate with front-end users, technical design engineers and pragmatic sales force members. · A tertiary qualification in a relevant science stream (eg. Medical Laboratory Science) · Practical experience in bench-level histopathology, including clinical immunohistochemistry · Demonstrated use of automated scientific instrumentation BS/ MS in Life Sciences and ASCP Certification Interest in using PC based software and maintenance of automated scientific instrumentation will be highly advantageous. In addition, you are extremely detailed and analytically minded with a strong sense of customer focus. Interested parties should forward a resume to resume@scigenium.com Send to : SciGenium PO 380916 Cambridge, MA 02238 From Malcolm.McCallum <@t> tamut.edu Wed Jan 25 13:48:09 2006 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Wed Jan 25 13:50:54 2006 Subject: [Histonet] Wanted : Field Support Specialist - Histopathology, TEXAS Message-ID: I am just wanting to clarify the employment situation for histotechs. How hard is it to get vacancies filled? Malcolm McCallum A&M_Texarkana -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Rynak Sent: Wednesday, January 25, 2006 1:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Wanted : Field Support Specialist - Histopathology, TEXAS Field Support Specialist - Histopathology SciGenium's Client is experiencing tremendous growth within their North American operations. This growth is due to the recent launch of their FDA approved automated histology instrumentation system, and the growing antibody and reagent consumable product line for the histology market. Due to the new launch of these products they are currently looking for Field Support Specialist - Histopathology, for Southwest Region (TX Houston or Dallas ) Based in Texas, this position provides technical applications support for our next generation range of automated histopathology products. This is a very hands on role where your key tasks will include: * Building an in-depth understanding of our new product technologies and their applications in a diagnostic histopathology environment; * Customer liaison by providing scientific and laboratory expertise for in-field installations, training and trouble-shooting; * Customer support Help-line for remote problem solving; * Designing and performing experiments to investigate and solve tough technical applications problems; and * Preparing problem resolution reports and imparting your findings and knowledge to internal and field based Product Engineers and Technical Managers. To be successful in this role you will possess: * Excellent problem solving and analytical skills * The ability to communicate with front-end users, technical design engineers and pragmatic sales force members. * A tertiary qualification in a relevant science stream (eg. Medical Laboratory Science) * Practical experience in bench-level histopathology, including clinical immunohistochemistry * Demonstrated use of automated scientific instrumentation BS/ MS in Life Sciences and ASCP Certification Interest in using PC based software and maintenance of automated scientific instrumentation will be highly advantageous. In addition, you are extremely detailed and analytically minded with a strong sense of customer focus. Interested parties should forward a resume to resume@scigenium.com Send to : SciGenium PO 380916 Cambridge, MA 02238 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kjsavage <@t> buffalo.edu Wed Jan 25 14:02:37 2006 From: kjsavage <@t> buffalo.edu (kjsavage@buffalo.edu) Date: Wed Jan 25 14:02:47 2006 Subject: [Histonet] methacarn problem solved Message-ID: <1138219357.43d7d95d8710a@mail3.buffalo.edu> Hi all, First, "thank you" to everyone who responded to my please for help with the methacarn solution. The problem was with the chloroform. A chemist friend of mine clued me in to the problem. Most chloroform comes with ethanol added as a perservative. But there are some with amylene as the preservative instead. Sometimes they do not list it as an additive on the bottle, and you only find out when you call the company. Filtering it out is an option. But I guess now that I know what was happening I will just get the choloroform without amylene. Thanks again to all for the helpful suggestions. Kathy Savage, PhD Dept. of Exercise and Nutrition Sciences SUNY-University at Buffalo From pathrm35 <@t> adelphia.net Wed Jan 25 14:34:14 2006 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Wed Jan 25 14:34:25 2006 Subject: [Histonet] seeking employment Message-ID: <29876775.1138221254204.JavaMail.root@web12.mail.adelphia.net> Histotechnologist seeking a lead tech or assistant supervisor position. Willing to relocate and work any shift. I prefer private, high volume labs and specialize in dermpath and IHC. Thanks in advance. Ron Martin, BS, HT (ASCP) HTL, QIHC From pruegg <@t> ihctech.net Wed Jan 25 15:40:20 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Wed Jan 25 15:40:36 2006 Subject: [Histonet] M2A Message-ID: <200601252140.k0PLeJjH003601@chip.viawest.net> has anyone used an antibody M2A Antigen (Lymphatic Endothelial Cell Marker) for either swine or ovine tissue? It works in human, but not canine, swine or ovine. I need swine or ovine. Thanks, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From YuJ2 <@t> upmc.edu Wed Jan 25 15:55:44 2006 From: YuJ2 <@t> upmc.edu (Yu, Jian) Date: Wed Jan 25 15:55:57 2006 Subject: [Histonet] cytochrome c IHC or IF on mouse small intestine Message-ID: <2554B4CA518D504A81E6908E39478A6A131FCD79@1upmc-msx10.isdip.upmc.edu> Thanks a lot for the great suggestions I got from several of you on the histopathology of mouse small intestine. There are a lot published results of cytochorme c release detected by IHC and IF in mouse liver and brain, but hardly anything in small intestine. Does anyone have a good protocol for cytochrome c release IHC or IF in mouse small intestine that you can share? Or it does not work at all? Thanks again!!! ******************************************************** Jian Yu, Ph. D. Assistant Professor of Pathology University of Pittsburgh Cancer Institute Hillman Cancer Center Research Pavilion Office suite 2.26h, Laboratory 2.43 5117 Centre Avenue, Pittsburgh, PA 15213 Phone : 412-623-7786, (Lab) 412-623-3255 Fax: 412-623-7778 Email: yuj2@upmc.edu ******************************************************** From dmarsha3 <@t> utmem.edu Wed Jan 25 16:00:43 2006 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Wed Jan 25 16:00:54 2006 Subject: [Histonet] RE: GCK-like kinase(GLK) References: Message-ID: <005c01c621fa$c98e3ac0$f623c084@DanaM> http://www.genecards.org/cgi-bin/carddisp?MAP4K4&search=GCK-like+kinase&suff=txt this is a link to Weizman Institute's GeneCards (www.genecards.org) i entered GCK-like kinase and as best i can tell it believes that is the same as map kinase kinase kinase kinase 4? regardless, if that is not the right one, you might go ahead and try to find it on there using your own criteria. as you scroll down the page you will see expression as reported using any number of methodologies as well as links out with more information about the expression data. there is also a site called iHOP that has tons of information about proteins. dana ----- Original Message ----- From: "Edwards, R.E." To: Sent: Wednesday, January 25, 2006 9:29 AM Subject: [Histonet] RE: GCK-like kinase(GLK) --Anyone have any idea of the distribution of GLK in normal human tissues, unable to find anything in Pubmed. Thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER..U.K..... _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Jan 25 16:08:32 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Jan 25 16:08:51 2006 Subject: [Histonet] M2A In-Reply-To: <200601252140.k0PLeJjH003601@chip.viawest.net> References: <200601252140.k0PLeJjH003601@chip.viawest.net> Message-ID: <6.0.0.22.1.20060125150737.01b4a340@gemini.msu.montana.edu> Patsy, Try this website, a new one for antibody location. www.exactantigen.com Good luck At 02:40 PM 1/25/2006, you wrote: >has anyone used an antibody M2A Antigen (Lymphatic Endothelial Cell Marker) Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From stephanie.brusig <@t> weyerhaeuser.com Wed Jan 25 16:18:32 2006 From: stephanie.brusig <@t> weyerhaeuser.com (Brusig, Stephanie) Date: Wed Jan 25 16:18:45 2006 Subject: [Histonet] Sakura Tissue Tek II Message-ID: <16E971922EDF9A4B9E8D3216374EDD49766C30@wafedixm12.corp.weyer.pri> Does any one have a operators manual they don't need for a Tissue Tek II? I don't feel like paying the $125 from Sukura. Thanks, ~Stephanie Brusig Weyerhaeuser Company Propagation of High Value Trees WTC-1B10 253.924.6518 From Lchausse <@t> nmh.org Wed Jan 25 16:50:40 2006 From: Lchausse <@t> nmh.org (Chaussey, Leslie) Date: Wed Jan 25 16:51:07 2006 Subject: [Histonet] Positions available Message-ID: Northwestern Memorial Hospital, consistently recognized as the most preferred hospital in Chicago, is seeking new team members. We hire the 'Best People' who are dedicated to being a part of the 'Best Patient Experience!' At Northwestern Memorial Hospital, we're setting new standards of care. We're also creating career opportunities that allow you to share your talent, develop new skills, and reach all of your goals. We're one of the best hospitals in the world because we support the efforts, the ideas, and the initiatives of all of our people. We invite you to join us. We are currently seeking qualified candidates for our Histology, Gross Room, and Electron Microscopy/Renal Laboratories openings. For more information and to submit your resume, please visit our website, www.nmh.org . AA/EOE ----------------------------------------- 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 KDwyer3322 <@t> aol.com Wed Jan 25 17:07:43 2006 From: KDwyer3322 <@t> aol.com (KDwyer3322@aol.com) Date: Wed Jan 25 17:07:54 2006 Subject: [Histonet] Texas Society for Histotechnology 2006 Program Message-ID: <1a0.44b03f7c.31095ebf@aol.com> To all: The Texas Society for Histotechnology will be having it's 2006 Symposium/Convention March 31-April 2, 2006 in Corpus Christi, Texas at the Omni Hotel. For a program or more information contact: Judy Webb - 817927-1024 or Veronica Davis -972-579-8291 Friday, March 31, 2006 10:00am-5:00 p.m. NSH Sponsored Workshop HT (ASCP) Examination Readiness - Glenda Hoye BS, HT (ASCP) Saturday, April 1, 2006 8-11:30 a.m. A.M. Workshops #1 Reagent Alcohol-Can't Drink It-So What is It? Pam Marcum HT(ASCP) #2 Technical Immunohistochemistry: Achieving Reliability and Reproducibility of Immunostains Rodney Miller, M.D. #3 Thinking LEAN in Histology R. Stephens HT(ASCP) QIHC #4. ASCP Certification Maintenance Program Evelyn Sandberg, HT(ASCP)HTL Saturday, April 1, 2006 1:00-4:30 p.m. P.M. Workshops #5 IHC Mathematics in the Laboratory Joel Martinez /Fatima Natar #6 MonKey Business-the Key to Delegation Jan Gardner ,MBA,HT(ASCP)/ Judi Stasko, BS, CLT #7 ISH/FISH Theory and Application Noemi Sebastiao #8. Gross Tissue Examination Charles Embrey,BS, HT(ASCP), PA(ASCP) Sunday,April 2, 2006 8:00-11:30 a.m. AM Workshops #9 CSI: Corpus Christi, Case Study Investigations Mike Reichenbach, HT (ASCP) QIHC Debra Flynn, HT(ASCP) QIHC #10 Ready or Not Here It Comes: Microwave Technology Donna Willis, HT(ASCP) HTL #11 Do You Know What Your Laboratory Workflow Is? Ritu Ward, MT(ASCP) MAOM #12. Making Job Descriptions Work for You Jan Gardner, MBA, HT (ASCP)/ Judi Stasko, BS, CLT From n.cragg <@t> epistem.co.uk Thu Jan 26 03:07:06 2006 From: n.cragg <@t> epistem.co.uk (Nicola Cragg) Date: Thu Jan 26 03:07:19 2006 Subject: [Histonet] Quantitative Image Analysis Systems - -Any recommendations? Message-ID: Hi All, Has anyone got any recommendations and/or advice on image analysis systems? We are looking to buy a new image analysis system to quantify IHC and perform histometric measurements (although this may require a second system). Does anyone have a system they can recommend? Or has anyone looked into this field and can offer any advice. Up to now, we've looked into Chromavision, Zeiss & Ariol. Thanks in advance, Nicola Cragg EpiStem Ltd. Manchester, UK From jqb7 <@t> cdc.gov Thu Jan 26 05:00:40 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Jan 26 04:59:31 2006 Subject: [Histonet] technique (Gimenez?) for rickettsiae Message-ID: Hi all: Can anyone recommend a good technique for demonstration of rickettsiae in FFPE tissue ? Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov From FAILM <@t> musc.edu Thu Jan 26 06:30:30 2006 From: FAILM <@t> musc.edu (Mildred Fail) Date: Thu Jan 26 06:32:38 2006 Subject: [Histonet] technique (Gimenez?) for rickettsiae Message-ID: Jeanine, Do both a Giemsa and Himenez, rickettsiae is + in both Rena Fail Rena Fail >>> "Bartlett, Jeanine" 01/26/06 06:00AM >>> Hi all: Can anyone recommend a good technique for demonstration of rickettsiae in FFPE tissue ? Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> cdc.gov Thu Jan 26 07:05:11 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Jan 26 07:04:38 2006 Subject: [Histonet] technique (Gimenez?) for rickettsiae Message-ID: They don't like the Giemsa for this: they want something that will make the bugs pop out at you. Jeanine Bartlett, HT(ASCP) (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: Mildred Fail [mailto:FAILM@musc.edu] Sent: Thursday, January 26, 2006 7:31 AM To: Bartlett, Jeanine; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] technique (Gimenez?) for rickettsiae Jeanine, Do both a Giemsa and Himenez, rickettsiae is + in both Rena Fail Rena Fail >>> "Bartlett, Jeanine" 01/26/06 06:00AM >>> Hi all: Can anyone recommend a good technique for demonstration of rickettsiae in FFPE tissue ? Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From FAILM <@t> musc.edu Thu Jan 26 07:10:47 2006 From: FAILM <@t> musc.edu (Mildred Fail) Date: Thu Jan 26 07:12:50 2006 Subject: [Histonet] CORRECTION Message-ID: gimenez and giemsa Rena Fail From kmerriam2003 <@t> yahoo.com Thu Jan 26 07:45:31 2006 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Thu Jan 26 07:45:41 2006 Subject: [Histonet] Quantitative Image Analysis Systems - -Any recommendations? In-Reply-To: Message-ID: <20060126134531.15492.qmail@web50306.mail.yahoo.com> We have an Aperio system (http://www.aperio.com/), it scans the entire slide at a 20X mag. The resolution is absolutely amazing, we compared all of the major brands mentioned below and this was by far the best system. It also comes with some nice image analysis software that can do pixel count, nuclear, membrane and they can be modified to fit your specific needs. We have the T2 system that can scan 120 slides at a time. It is definitely worth looking into. Kim Merriam Novartis Cambridge, MA Nicola Cragg wrote: Hi All, Has anyone got any recommendations and/or advice on image analysis systems? We are looking to buy a new image analysis system to quantify IHC and perform histometric measurements (although this may require a second system). Does anyone have a system they can recommend? Or has anyone looked into this field and can offer any advice. Up to now, we've looked into Chromavision, Zeiss & Ariol. Thanks in advance, Nicola Cragg EpiStem Ltd. Manchester, UK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. From rjbuesa <@t> yahoo.com Thu Jan 26 07:55:17 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Jan 26 07:55:26 2006 Subject: [Histonet] technique (Gimenez?) for rickettsiae In-Reply-To: Message-ID: <20060126135517.46518.qmail@web61219.mail.yahoo.com> Jeanine: Between 1911 and 1945 there were 21 methods/variants developed for rickettsiaeThe one by Wolbach (1919) stain nuclei and Rickettsiae blue and cytoplasm red. L?pine's (1932) stain Rickettsiae red in pale brown cells. Many others are variations of red agains blues (similar to Giemsa used for this purpose). I prefer Wolbach's. Hope this will help. Ren? J. "Bartlett, Jeanine" wrote: Hi all: Can anyone recommend a good technique for demonstration of rickettsiae in FFPE tissue ? Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. From jqb7 <@t> cdc.gov Thu Jan 26 08:03:51 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Jan 26 08:07:03 2006 Subject: [Histonet] technique (Gimenez?) for rickettsiae Message-ID: We use the Wolbach's Giemsa but he doesn't care for it for this purpose. Thanks. Jeanine Bartlett, HT(ASCP) (404) 639-3590 jeanine.bartlett@cdc.hhs.gov _____ From: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: Thursday, January 26, 2006 8:55 AM To: Bartlett, Jeanine; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] technique (Gimenez?) for rickettsiae Jeanine: Between 1911 and 1945 there were 21 methods/variants developed for rickettsiaeThe one by Wolbach (1919) stain nuclei and Rickettsiae blue and cytoplasm red. L?pine's (1932) stain Rickettsiae red in pale brown cells. Many others are variations of red agains blues (similar to Giemsa used for this purpose). I prefer Wolbach's. Hope this will help. Ren? J. "Bartlett, Jeanine" wrote: Hi all: Can anyone recommend a good technique for demonstration of rickettsiae in FFPE tissue ? Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____ Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. From boycea <@t> mail.nih.gov Thu Jan 26 09:01:30 2006 From: boycea <@t> mail.nih.gov (Boyce, Amanda (NIH/NIAMS) [F]) Date: Thu Jan 26 09:01:47 2006 Subject: [Histonet] skeletal staining Message-ID: <72210BBE225D724FBDB0DF25C67B1A5305D99B@NIHCESMLBX9.nih.gov> I'm in the middle of doing alcian blue/alizarin red whole skeletal chick preps and I have a question. The protocol I've chosen uses 95% ethanol for the fixative. I've already fixed in PFA. Is it possible to continue with the protocol? Are there any steps I should add? Thanks, Amanda Boyce From ian.montgomery <@t> bio.gla.ac.uk Thu Jan 26 09:29:33 2006 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Thu Jan 26 09:29:55 2006 Subject: [Histonet] Advice. Message-ID: <7.0.1.0.2.20060126151836.03b48eb0@bio.gla.ac.uk> With cuts in staff I've been asked if I can teach the histology portion of a zoology class. Problem, one of the tissue I'll be using is insect abdomen with its chitin exoskeleton. Any hints and tips regarding this type of material, processing, embedding, sectioning, that sort of thing. Or, is it Mollifix time again? Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From mward <@t> wfubmc.edu Thu Jan 26 09:39:52 2006 From: mward <@t> wfubmc.edu (Martha Ward) Date: Thu Jan 26 09:40:21 2006 Subject: [Histonet] Hep Par 1 antibody Message-ID: <61135F0455D33347B5AAE209B903A3041209894B@EXCHVS2.medctr.ad.wfubmc.edu> I would like to thank everyone that responded to my question. I hope to get it up and running quickly. Martha Ward Wake Forest University Baptist Medical Center From Barry.R.Rittman <@t> uth.tmc.edu Thu Jan 26 09:48:26 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Thu Jan 26 09:49:27 2006 Subject: [Histonet] skeletal staining Message-ID: No problem Just make sure that you remove digestive tract as it tends to bind alizarin. Barry ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Boyce, Amanda (NIH/NIAMS) [F] Sent: Thu 1/26/2006 9:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] skeletal staining I'm in the middle of doing alcian blue/alizarin red whole skeletal chick preps and I have a question. The protocol I've chosen uses 95% ethanol for the fixative. I've already fixed in PFA. Is it possible to continue with the protocol? Are there any steps I should add? Thanks, Amanda Boyce _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Thu Jan 26 09:52:48 2006 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Thu Jan 26 09:54:51 2006 Subject: [Histonet] Zinc Formalin Message-ID: It's been a few years since I've worked with zinc formalin. But if my memory serves me right (ha ha), you should wash the tissue thoroughly befoe going back into 10%NBF. Something about the compatibility of zinc with the phosphate buffer. Fred >>> "Laurie Colbert" 01/24/06 06:12PM >>> Our formalin supplier mistakenly shipped us pre-filled containers containing zinc formalin instead of 10% NBF. Before I noticed this, three cases had been distributed throughout the hospital (most likely to Surgery). So, how is this going to affect my tissue? Is there any post-treatment (such as removing mercury pigment on B-5 fixed tissue) that I need to perform? Thanks in advance for any info you can provide. Laurie Colbert _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Thu Jan 26 09:54:14 2006 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Thu Jan 26 09:55:07 2006 Subject: [Histonet] H.pylori immunos on gastric biopsies Message-ID: I would have bet $10 I wouldn't have seen that phrase today. >>> "Marshall Terry Dr, Consultant Histopathologist" 01/25/06 06:54AM >>> Yes, but a turd is easier to bottle. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:kemlo.rogerson@waht.swest.nhs.uk] Sent: 25 January 2006 09:02 To: Marshall Terry Dr, Consultant Histopathologist; Smith, Jeffery D. (HSC); Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies Isn't there a breath test too? Kemlo Rogerson Weston Super Mare Cell Path Manager -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, January 24, 2006 5:09 PM To: Smith, Jeffery D. (HSC); Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies H. pylori is a pathogen. It does not sit in the stomach behaving itself. Its presence is an indication for treatment, but its absence is *not* an indication for treatment directed against it, irrespective of how inflamed the stomach looks. The best histologic determinant of H. pylori is an immunostain. However, in most cases, this is all rather academic, as a quick stool test is available, directed against H. pylori antigen, so in most cases, we are really trying to determine something better done by other means. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Smith, Jeffery D. (HSC) [mailto:Jeffery-Smith@ouhsc.edu] Sent: 24 January 2006 16:18 To: Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies I have to somewhat agree. Immunos for H. pylori seems to be overkill. We routinely stain for H. pylori using a Giemsa on all GI biopsies. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bill Blank Sent: Tue 1/24/2006 9:29 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies IMO, we take this detecting H. pylori thing too seriously. A rational physician treats the patient and not any lab test. If I had symptoms of H. pylori, if my stomach looked inflamed, I would want to be treated for H. pylori irregardless of their presence on a biopsy. (Well, maybe as there is some evidence that treating antral H. pylori may increase the risk of GE junction adenocarcinoma) I consider immunos on gastric biopsies to be overkill and a waste of health care dollars. BIll At 7:53 AM -0600 1/24/06, GUTIERREZ, JUAN wrote: >We do immunos on all gastric biopsies. We had some cases that looked >negative by giemsa stain, yet the clinical hx suggested otherwise. When >we ran the immunos on these so called negatives, we were shocked to see >the amount of organisms we had missed. Do you want to take that chance? >I always ask myself: what if this was my biopsy? Something to think >about. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Thu Jan 26 10:08:22 2006 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Thu Jan 26 10:07:05 2006 Subject: [Histonet] H.pylori immunos on gastric biopsies Message-ID: Then you would have lost:-) (It's probably the first time in recorded history that is *has* been used.) Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Fred Underwood [mailto:funderwood@mcohio.org] Sent: 26 January 2006 15:54 To: Histonet@lists.utsouthwestern.edu; bill501@mindspring.com; Jeffery-Smith@ouhsc.edu; Marshall Terry Dr, Consultant Histopathologist; kemlo.rogerson@waht.swest.nhs.uk Subject: RE: [Histonet] H.pylori immunos on gastric biopsies I would have bet $10 I wouldn't have seen that phrase today. >>> "Marshall Terry Dr, Consultant Histopathologist" 01/25/06 06:54AM >>> Yes, but a turd is easier to bottle. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:kemlo.rogerson@waht.swest.nhs.uk] Sent: 25 January 2006 09:02 To: Marshall Terry Dr, Consultant Histopathologist; Smith, Jeffery D. (HSC); Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies Isn't there a breath test too? Kemlo Rogerson Weston Super Mare Cell Path Manager -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, January 24, 2006 5:09 PM To: Smith, Jeffery D. (HSC); Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies H. pylori is a pathogen. It does not sit in the stomach behaving itself. Its presence is an indication for treatment, but its absence is *not* an indication for treatment directed against it, irrespective of how inflamed the stomach looks. The best histologic determinant of H. pylori is an immunostain. However, in most cases, this is all rather academic, as a quick stool test is available, directed against H. pylori antigen, so in most cases, we are really trying to determine something better done by other means. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Smith, Jeffery D. (HSC) [mailto:Jeffery-Smith@ouhsc.edu] Sent: 24 January 2006 16:18 To: Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies I have to somewhat agree. Immunos for H. pylori seems to be overkill. We routinely stain for H. pylori using a Giemsa on all GI biopsies. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bill Blank Sent: Tue 1/24/2006 9:29 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies IMO, we take this detecting H. pylori thing too seriously. A rational physician treats the patient and not any lab test. If I had symptoms of H. pylori, if my stomach looked inflamed, I would want to be treated for H. pylori irregardless of their presence on a biopsy. (Well, maybe as there is some evidence that treating antral H. pylori may increase the risk of GE junction adenocarcinoma) I consider immunos on gastric biopsies to be overkill and a waste of health care dollars. BIll At 7:53 AM -0600 1/24/06, GUTIERREZ, JUAN wrote: >We do immunos on all gastric biopsies. We had some cases that looked >negative by giemsa stain, yet the clinical hx suggested otherwise. When >we ran the immunos on these so called negatives, we were shocked to see >the amount of organisms we had missed. Do you want to take that chance? >I always ask myself: what if this was my biopsy? Something to think >about. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Thu Jan 26 10:06:24 2006 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Thu Jan 26 10:07:08 2006 Subject: [Histonet] skeletal staining Message-ID: <5784D843593D874C93E9BADCB87342AB916B53@tpiserver03.Coretech-holdings.com> A customer has asked the following question. Can anybody help? "Can you recommend on what corner I need establish a knife when I'm working with Vibratome 8000, if I work with Super Mega Cassette and using Feather mikrotome blade S35". Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R Sent: Thursday, January 26, 2006 9:48 AM To: Boyce, Amanda (NIH/NIAMS) [F]; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] skeletal staining No problem Just make sure that you remove digestive tract as it tends to bind alizarin. Barry ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Boyce, Amanda (NIH/NIAMS) [F] Sent: Thu 1/26/2006 9:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] skeletal staining I'm in the middle of doing alcian blue/alizarin red whole skeletal chick preps and I have a question. The protocol I've chosen uses 95% ethanol for the fixative. I've already fixed in PFA. Is it possible to continue with the protocol? Are there any steps I should add? Thanks, Amanda Boyce _______________________________________________ Histonet mailing list Histonet@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 Barry.R.Rittman <@t> uth.tmc.edu Thu Jan 26 10:08:14 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Thu Jan 26 10:08:18 2006 Subject: [Histonet] Advice. Message-ID: Ian I did a lot of sectioning of dragonfly larvae a long time ago and tried all sorts of processing procedures. The best way that I found to keep tissue soft enough for paraffin wax sections was to use a chloral hydrate: phenol mixture. After fixation and dehydration to absolute ethanol, soaked in a mixture of equal parts of chloral hydrate:phenol (heated gently to allow mixing of these two). Then used chloroform intermediary agent, then paraffin etc. Also found that if soaked the insects in equal parts of chloroform and paraffin at least overnight at room temperature that this significantly cut down the time needed in molten paraffin wax. I realize that chloral hydrate is not that easy to obtain (was 38 years ago that I did this) but I found this the best method. Another method that has been used is to use N-butyl alcohol or isopropyl alcohol for dehydration as these have a less hardening effect. Have not used N-butyl alcohol but have used isoproyl and this is somewhat miscible with paraffin wax. This method was an improvement over routine wax processing but not as good as chloral hydrate:phenol softening. Hope that this helps. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ian Montgomery Sent: Thu 1/26/2006 9:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Advice. With cuts in staff I've been asked if I can teach the histology portion of a zoology class. Problem, one of the tissue I'll be using is insect abdomen with its chitin exoskeleton. Any hints and tips regarding this type of material, processing, embedding, sectioning, that sort of thing. Or, is it Mollifix time again? Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Thu Jan 26 10:31:38 2006 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Thu Jan 26 10:32:19 2006 Subject: [Histonet] RE: Microtome configuration Message-ID: <5784D843593D874C93E9BADCB87342AB013069DF@tpiserver03.Coretech-holdings.com> Sorry, last send had wrong subject line Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles Scouten Sent: Thursday, January 26, 2006 10:06 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] skeletal staining A customer has asked the following question. Can anybody help? "Can you recommend on what corner I need establish a knife when I'm working with Vibratome 8000, if I work with Super Mega Cassette and using Feather mikrotome blade S35". Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R Sent: Thursday, January 26, 2006 9:48 AM To: Boyce, Amanda (NIH/NIAMS) [F]; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] skeletal staining No problem Just make sure that you remove digestive tract as it tends to bind alizarin. Barry ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Boyce, Amanda (NIH/NIAMS) [F] Sent: Thu 1/26/2006 9:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] skeletal staining I'm in the middle of doing alcian blue/alizarin red whole skeletal chick preps and I have a question. The protocol I've chosen uses 95% ethanol for the fixative. I've already fixed in PFA. Is it possible to continue with the protocol? Are there any steps I should add? Thanks, Amanda Boyce _______________________________________________ Histonet mailing list Histonet@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 PMonfils <@t> Lifespan.org Thu Jan 26 10:33:46 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Thu Jan 26 10:33:57 2006 Subject: [Histonet] Zinc Formalin Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717658@lsexch.lsmaster.lifespan.org> That's correct, Fred. I don't recall every having to transfer tissue from zinc formalin into NBF; however, zinc phosphate is insoluble in water at neutral pH. Therefore introducing phosphate ions into a system which already contains free zinc ions will result in precipitation of solid zinc phosphate, in this case within the tissue specimen. All unbound zinc should be washed out of the tissue before placing the tissue in a phosphate-buffered solution. This would include not only NBF but also PBS or other phosphate-based buffers. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Fred > Underwood > Sent: Thursday, January 26, 2006 7:52 AM > To: laurie.colbert@huntingtonhospital.com; > histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Zinc Formalin > > It's been a few years since I've worked with zinc formalin. But if my > memory serves me right (ha ha), you should wash the tissue thoroughly > befoe going back into 10%NBF. Something about the compatibility of zinc > with the phosphate buffer. > > Fred > > >>> "Laurie Colbert" 01/24/06 > 06:12PM >>> > Our formalin supplier mistakenly shipped us pre-filled containers > containing zinc formalin instead of 10% NBF. Before I noticed this, > three cases had been distributed throughout the hospital (most likely to > Surgery). So, how is this going to affect my tissue? Is there any > post-treatment (such as removing mercury pigment on B-5 fixed tissue) > that I need to perform? > > Thanks in advance for any info you can provide. > > 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 > > From laurie.colbert <@t> huntingtonhospital.com Thu Jan 26 10:56:44 2006 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Thu Jan 26 10:56:55 2006 Subject: [Histonet] Zinc Formalin Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628005653978@EXCHANGE1.huntingtonhospital.com> Thanks for all of your responses to my zinc formalin question. I think we got most of the containers back from Surgery, so hopefully this won't even be an issue. Laurie Colbert From gcallis <@t> montana.edu Thu Jan 26 11:55:59 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Jan 26 11:56:22 2006 Subject: [Histonet] Advice. In-Reply-To: <7.0.1.0.2.20060126151836.03b48eb0@bio.gla.ac.uk> References: <7.0.1.0.2.20060126151836.03b48eb0@bio.gla.ac.uk> Message-ID: <6.0.0.22.1.20060126105151.01b6ddc0@gemini.msu.montana.edu> For some reason, I have a chapter copied from a book titled Animal and Insect Histotechnology, no author - all I have is a chapter from this book authored by Debra A. McElroy - but it contained some words of wisdom for insects. Fixation NBF, Bouins, or Zenkers - after fixation, soak fixed specimens in 4% phenol (4 ml melted phenol in 96 ml 80% ethanol) for 24 hours to soften the chitnous exoskeletaons then process into paraffin. If you try this, please keep us posted on results. At 08:29 AM 1/26/2006, you wrote: > With cuts in staff I've been asked if I can teach the histology > portion of a zoology class. Problem, one of the tissue I'll be using is > insect abdomen with its chitin exoskeleton. Any hints and tips regarding > this type of material, processing, embedding, sectioning, that sort of > thing. Or, is it Mollifix time again? > >Dr. Ian Montgomery, >Histotechnology, >IBLS Support Services, >Graham Kerr Building, >Institute of Biomedical & Life Sciences, >University of Glasgow, >Glasgow, >G12 8QQ. >Tel: 0141 339 8855 >Office: 4652 >Lab: 6644. >Pager: 07623 975451 >e-mail: ian.montgomery@bio.gla.ac.uk > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From Rcartun <@t> harthosp.org Thu Jan 26 11:56:05 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Thu Jan 26 11:56:43 2006 Subject: [Histonet] Hep Par 1 antibody Message-ID: We use Dako's mouse monoclonal (clone OCH1E5) at a dilution of 1:200 for 30' (following HIER) with their EnVision+/DAB+ detection. To be perfectly honest, although normal hepatocytes stain well, I have been disappointed with it's sensitivity in labeling hepatocellular carcinomas. 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 >>> "Martha Ward" 01/25/06 01:22PM >>> I have approached by our Pathologists about working up the Hep Par 1 antibody and I was wondering if anyone out there is currently doing it, and if so, could you recommend a vendor, conditions, etc. Thanks for much in advance for any help you can give me. Martha Ward, MT (ASCP) QIHC Molecular Diagnostics Lab Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From twebster <@t> nmcinc.org Thu Jan 26 12:17:08 2006 From: twebster <@t> nmcinc.org (Tim Webster) Date: Thu Jan 26 12:17:23 2006 Subject: [Histonet] SUSPECT: RE: SUSPECT: Histonet Digest, Vol 26, Issue 31 Message-ID: Yes Ma'am Tim Webster Histology Specialist Northwestern Medical Center 133 Fairfield Street St Albans, VT 05478 (802) 524-1070 x4349 (Dept. & Voice mail) twebster@nmcinc.org Statement of Confidentiality This message and any attachments are from NMC and intended only for the addressee(s). The information contained may include privileged or other confidential information. Unauthorized review, forwarding, printing, copying or distribution is strictly prohibited. Any opinions expressed are those of the author and not necessarily those of Norhtwestern Medical Center. If you should receive this message in error, please delete it and notify the sender. Thank you. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Thursday, January 26, 2006 11:25 AM To: histonet@lists.utsouthwestern.edu Subject: SUSPECT: Histonet Digest, Vol 26, Issue 31 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Hep Par 1 antibody (Martha Ward) 2. Re: Zinc Formalin (John A. Kiernan) 3. Re: (no subject) (John A. Kiernan) 4. RE: Hep Par 1 antibody (GUTIERREZ, JUAN) 5. RE: Hep Par 1 antibody (Sebree Linda A.) 6. Re: Qu:Assesment of embryonic BBB defect (John A. Kiernan) 7. RE: Helico Immuno (Cazares, Ruth) 8. Wanted : Field Support Specialist - Histopathology, TEXAS (John Rynak) 9. RE: Wanted : Field Support Specialist - Histopathology, TEXAS (Malcolm McCallum) 10. methacarn problem solved (kjsavage@buffalo.edu) 11. seeking employment (pathrm35@adelphia.net) 12. M2A (Patsy Ruegg) 13. cytochrome c IHC or IF on mouse small intestine (Yu, Jian) 14. Re: RE: GCK-like kinase(GLK) (Dana Marshall) 15. Re: M2A (Gayle Callis) 16. Sakura Tissue Tek II (Brusig, Stephanie) 17. Positions available (Chaussey, Leslie) 18. Texas Society for Histotechnology 2006 Program (KDwyer3322@aol.com) 19. Quantitative Image Analysis Systems - -Any recommendations? (Nicola Cragg) 20. technique (Gimenez?) for rickettsiae (Bartlett, Jeanine) 21. Re: technique (Gimenez?) for rickettsiae (Mildred Fail) 22. RE: technique (Gimenez?) for rickettsiae (Bartlett, Jeanine) 23. CORRECTION (Mildred Fail) 24. Re: Quantitative Image Analysis Systems - -Any recommendations? (Kim Merriam) 25. Re: technique (Gimenez?) for rickettsiae (Rene J Buesa) 26. RE: technique (Gimenez?) for rickettsiae (Bartlett, Jeanine) 27. skeletal staining (Boyce, Amanda (NIH/NIAMS) [F]) 28. Advice. (Ian Montgomery) 29. Hep Par 1 antibody (Martha Ward) ---------------------------------------------------------------------- Message: 1 Date: Wed, 25 Jan 2006 13:22:22 -0500 From: "Martha Ward" Subject: [Histonet] Hep Par 1 antibody To: Message-ID: <61135F0455D33347B5AAE209B903A3041206E7D6@EXCHVS2.medctr.ad.wfubmc.edu> Content-Type: text/plain; charset="us-ascii" I have approached by our Pathologists about working up the Hep Par 1 antibody and I was wondering if anyone out there is currently doing it, and if so, could you recommend a vendor, conditions, etc. Thanks for much in advance for any help you can give me. Martha Ward, MT (ASCP) QIHC Molecular Diagnostics Lab Wake Forest University Baptist Medical Center ------------------------------ Message: 2 Date: Wed, 25 Jan 2006 13:26:22 -0500 From: "John A. Kiernan" Subject: Re: [Histonet] Zinc Formalin To: Laurie Colbert Cc: "Histonet \(E-mail\)" Message-ID: <43D7C2CE.54157A1A@uwo.ca> Content-Type: text/plain; charset=us-ascii Zinc-formalin mixtures do not contain mercuric chloride, so you will not need to do an iodine and thiosulphate treatment of the sections. Ask your supplier to tell you the composition of his "zinc formalin". Is it an acidic (zinc sulphate or chloride) or a neutral (zinc salicylate) formulation? Most zinc formalin mixtures are acidic, and it may make a difference if the anion is sulphate or chloride. The published literature is clouded by trade secrecy, as are the web sites of those who make the mixtures. The acidic zinc formalin principle is simple: combine a protein coagulant with a cross-linker. All the classical late 19th century fixatives did that, and more. Formalin pigment can be expected if tissues are stored for a long time in an acidic formaldehyde solution. Hope this helps. John Kiernan Anatomy, UWO London, Canada. ______________________________________ Laurie Colbert wrote: > > Our formalin supplier mistakenly shipped us pre-filled containers containing zinc formalin instead of 10% NBF. Before I noticed this, three cases had been distributed throughout the hospital (most likely to Surgery). So, how is this going to affect my tissue? Is there any post-treatment (such as removing mercury pigment on B-5 fixed tissue) that I need to perform? > > Thanks in advance for any info you can provide. > > Laurie Colbert > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Wed, 25 Jan 2006 13:29:06 -0500 From: "John A. Kiernan" Subject: Re: [Histonet] (no subject) To: kjsavage@buffalo.edu Cc: histonet@lists.utsouthwestern.edu Message-ID: <43D7C372.2EC99A99@uwo.ca> Content-Type: text/plain; charset=us-ascii Dear kjsavage@buffalo.edu Methacarn is easy to make. It contains nothing that could form a precipitate. Your email indicates that you have all the ingredients (chloroform, methanol, acetic acid). This is a non-aqueous coagulant fixative, differing from Carnoy only in having methanol instead of ethanol. Fixatives of this kind contain no water. Cloudiness or a precipitate might happen if an ingredient is wrong or if water gets into the mixture. Are your methanol and acetic acid 95-100%? The advice to avoid plastic was good, because chloroform attacks some plastic ware. John Kiernan Anatomy, UWO London, Canada ______________________________________________________ kjsavage@buffalo.edu wrote: > > Greetings all, > > I am having a difficult time trying to make methacarn fixative. I have > searched the list archives and found a posting in May 2005 from someone > that was having the same problem I am facing: a white precipitate > forming. In one of the responses to his posting it was suggested to > use glass only (pipets, vials etc...). I have done that, and still the > precipitate forms. I have also used all new reagents (chloroform, > methanol, acetic acid) and still the precipitate forms. Does anyone > have some more suggestions as to what is going wrong? Thanks, Kathy > > Kathy Savage, PhD > Dept. of Exercise and Nutrition Sciences > SUNY-University at Buffalo > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Wed, 25 Jan 2006 12:34:30 -0600 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] Hep Par 1 antibody To: "Martha Ward" , Message-ID: Content-Type: text/plain; charset="US-ASCII" We use DAKO's Hepatocyte antibody, clone OCH1E5. It's very easy to work up on the Ventana stainers. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Martha Ward Sent: Wednesday, January 25, 2006 12:22 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Hep Par 1 antibody I have approached by our Pathologists about working up the Hep Par 1 antibody and I was wondering if anyone out there is currently doing it, and if so, could you recommend a vendor, conditions, etc. Thanks for much in advance for any help you can give me. Martha Ward, MT (ASCP) QIHC Molecular Diagnostics Lab Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Wed, 25 Jan 2006 12:46:21 -0600 From: "Sebree Linda A." Subject: RE: [Histonet] Hep Par 1 antibody To: "Martha Ward" , Message-ID: Content-Type: text/plain; charset="US-ASCII" Martha, We use Biocare's Hepatocyte Specific Antibody at 1:50 with a polymer DAB kit on our Ventana automated immunostainers. Our pathologists are happy with it. Linda Sebree, HT(ASCP) University of Wisconsin Hospital & Clinics IHC/ISH Laboratory A4/204-3224 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Martha Ward Sent: Wednesday, January 25, 2006 12:22 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Hep Par 1 antibody I have approached by our Pathologists about working up the Hep Par 1 antibody and I was wondering if anyone out there is currently doing it, and if so, could you recommend a vendor, conditions, etc. Thanks for much in advance for any help you can give me. Martha Ward, MT (ASCP) QIHC Molecular Diagnostics Lab Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Wed, 25 Jan 2006 13:48:10 -0500 From: "John A. Kiernan" Subject: Re: [Histonet] Qu:Assesment of embryonic BBB defect To: Ali Moussavi Nik , histonet@lists.utsouthwestern.edu Message-ID: <43D7C7EA.CFA7A5D5@uwo.ca> Content-Type: text/plain; charset=iso-8859-1 Immunostain for a plasma protein such as albumin. It's important to fix the tissue quickly, or plasma will move out of the blood vessels. See Mori et al (1991) Lab. Invest. 64:345-351; Fabian (1992) J. Histochem. Cytochem. 40: 987-991; Loberg & Torvik (1992) Acta path. microbiol. scand. 100: 431-436. I've seen this artifact in human brain, with immunostaining for albumin, ceruloplasmin, orosomucoid and IgG (unpublished). The time of first formation of the BBB is still controversial (far too much literature to mention here) and may depend on the techniques used. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Ali Moussavi Nik wrote: > > Dear All > > Hi > > Dose any one knows how to asses the embryonic Blood Brain Barrier defect. > > I would like to see whether or not the Blood Brain Barrier of 18.5 mouse > embryo of a nock out strain is leaky. > > I will extremely thankful if someone gives me a practical protocol. > > > > > > > > ---------------------------------------------------------------------------- > ------ > > Seyed Ali Moussavi Nik D.V.M , PhD Student > > Department of Cell and Molecular Biology, > > Gothenburg University > > Box 462 > > S-405 30 G?teborg > > Sweden > > Phone: +46(031) 7733895 > > Fax: +46(031) 7733801 > > www.molbio.gu.se > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Wed, 25 Jan 2006 13:04:22 -0600 From: "Cazares, Ruth" Subject: [Histonet] RE: Helico Immuno To: Message-ID: <913FAC2B773C19488E26AE6572180FA50458A5CF@exch01.schosp.org> Content-Type: text/plain; charset="us-ascii" Thanks to all of you who responded to my inquiry on immunos for H. pylori. It really helps to get different opinions and perspectives to help one weigh the pros and cons. Sincerely, Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. ------------------------------ Message: 8 Date: Wed, 25 Jan 2006 14:30:35 -0500 From: "John Rynak" Subject: [Histonet] Wanted : Field Support Specialist - Histopathology, TEXAS To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format="flowed" Field Support Specialist - Histopathology SciGenium's Client is experiencing tremendous growth within their North American operations. This growth is due to the recent launch of their FDA approved automated histology instrumentation system, and the growing antibody and reagent consumable product line for the histology market. Due to the new launch of these products they are currently looking for Field Support Specialist - Histopathology, for Southwest Region (TX Houston or Dallas ) Based in Texas, this position provides technical applications support for our next generation range of automated histopathology products. This is a very hands on role where your key tasks will include: ? Building an in-depth understanding of our new product technologies and their applications in a diagnostic histopathology environment; ? Customer liaison by providing scientific and laboratory expertise for in-field installations, training and trouble-shooting; ? Customer support Help-line for remote problem solving; ? Designing and performing experiments to investigate and solve tough technical applications problems; and ? Preparing problem resolution reports and imparting your findings and knowledge to internal and field based Product Engineers and Technical Managers. To be successful in this role you will possess: ? Excellent problem solving and analytical skills ? The ability to communicate with front-end users, technical design engineers and pragmatic sales force members. ? A tertiary qualification in a relevant science stream (eg. Medical Laboratory Science) ? Practical experience in bench-level histopathology, including clinical immunohistochemistry ? Demonstrated use of automated scientific instrumentation BS/ MS in Life Sciences and ASCP Certification Interest in using PC based software and maintenance of automated scientific instrumentation will be highly advantageous. In addition, you are extremely detailed and analytically minded with a strong sense of customer focus. Interested parties should forward a resume to resume@scigenium.com Send to : SciGenium PO 380916 Cambridge, MA 02238 ------------------------------ Message: 9 Date: Wed, 25 Jan 2006 13:48:09 -0600 From: "Malcolm McCallum" Subject: RE: [Histonet] Wanted : Field Support Specialist - Histopathology, TEXAS To: "John Rynak" , Message-ID: Content-Type: text/plain; charset="us-ascii" I am just wanting to clarify the employment situation for histotechs. How hard is it to get vacancies filled? Malcolm McCallum A&M_Texarkana -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Rynak Sent: Wednesday, January 25, 2006 1:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Wanted : Field Support Specialist - Histopathology, TEXAS Field Support Specialist - Histopathology SciGenium's Client is experiencing tremendous growth within their North American operations. This growth is due to the recent launch of their FDA approved automated histology instrumentation system, and the growing antibody and reagent consumable product line for the histology market. Due to the new launch of these products they are currently looking for Field Support Specialist - Histopathology, for Southwest Region (TX Houston or Dallas ) Based in Texas, this position provides technical applications support for our next generation range of automated histopathology products. This is a very hands on role where your key tasks will include: * Building an in-depth understanding of our new product technologies and their applications in a diagnostic histopathology environment; * Customer liaison by providing scientific and laboratory expertise for in-field installations, training and trouble-shooting; * Customer support Help-line for remote problem solving; * Designing and performing experiments to investigate and solve tough technical applications problems; and * Preparing problem resolution reports and imparting your findings and knowledge to internal and field based Product Engineers and Technical Managers. To be successful in this role you will possess: * Excellent problem solving and analytical skills * The ability to communicate with front-end users, technical design engineers and pragmatic sales force members. * A tertiary qualification in a relevant science stream (eg. Medical Laboratory Science) * Practical experience in bench-level histopathology, including clinical immunohistochemistry * Demonstrated use of automated scientific instrumentation BS/ MS in Life Sciences and ASCP Certification Interest in using PC based software and maintenance of automated scientific instrumentation will be highly advantageous. In addition, you are extremely detailed and analytically minded with a strong sense of customer focus. Interested parties should forward a resume to resume@scigenium.com Send to : SciGenium PO 380916 Cambridge, MA 02238 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 10 Date: Wed, 25 Jan 2006 15:02:37 -0500 From: kjsavage@buffalo.edu Subject: [Histonet] methacarn problem solved To: histonet@lists.utsouthwestern.edu Message-ID: <1138219357.43d7d95d8710a@mail3.buffalo.edu> Content-Type: text/plain Hi all, First, "thank you" to everyone who responded to my please for help with the methacarn solution. The problem was with the chloroform. A chemist friend of mine clued me in to the problem. Most chloroform comes with ethanol added as a perservative. But there are some with amylene as the preservative instead. Sometimes they do not list it as an additive on the bottle, and you only find out when you call the company. Filtering it out is an option. But I guess now that I know what was happening I will just get the choloroform without amylene. Thanks again to all for the helpful suggestions. Kathy Savage, PhD Dept. of Exercise and Nutrition Sciences SUNY-University at Buffalo ------------------------------ Message: 11 Date: Wed, 25 Jan 2006 15:34:14 -0500 From: pathrm35@adelphia.net Subject: [Histonet] seeking employment To: histonet@lists.utsouthwestern.edu Message-ID: <29876775.1138221254204.JavaMail.root@web12.mail.adelphia.net> Content-Type: text/plain; charset=utf-8 Histotechnologist seeking a lead tech or assistant supervisor position. Willing to relocate and work any shift. I prefer private, high volume labs and specialize in dermpath and IHC. Thanks in advance. Ron Martin, BS, HT (ASCP) HTL, QIHC ------------------------------ Message: 12 Date: Wed, 25 Jan 2006 14:40:20 -0700 From: "Patsy Ruegg" Subject: [Histonet] M2A To: Message-ID: <200601252140.k0PLeJjH003601@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" has anyone used an antibody M2A Antigen (Lymphatic Endothelial Cell Marker) for either swine or ovine tissue? It works in human, but not canine, swine or ovine. I need swine or ovine. Thanks, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. ------------------------------ Message: 13 Date: Wed, 25 Jan 2006 16:55:44 -0500 From: "Yu, Jian" Subject: [Histonet] cytochrome c IHC or IF on mouse small intestine To: Message-ID: <2554B4CA518D504A81E6908E39478A6A131FCD79@1upmc-msx10.isdip.upmc.edu> Content-Type: text/plain; charset="us-ascii" Thanks a lot for the great suggestions I got from several of you on the histopathology of mouse small intestine. There are a lot published results of cytochorme c release detected by IHC and IF in mouse liver and brain, but hardly anything in small intestine. Does anyone have a good protocol for cytochrome c release IHC or IF in mouse small intestine that you can share? Or it does not work at all? Thanks again!!! ******************************************************** Jian Yu, Ph. D. Assistant Professor of Pathology University of Pittsburgh Cancer Institute Hillman Cancer Center Research Pavilion Office suite 2.26h, Laboratory 2.43 5117 Centre Avenue, Pittsburgh, PA 15213 Phone : 412-623-7786, (Lab) 412-623-3255 Fax: 412-623-7778 Email: yuj2@upmc.edu ******************************************************** ------------------------------ Message: 14 Date: Wed, 25 Jan 2006 16:00:43 -0600 From: "Dana Marshall" Subject: Re: [Histonet] RE: GCK-like kinase(GLK) To: "Edwards, R.E." , Message-ID: <005c01c621fa$c98e3ac0$f623c084@DanaM> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original http://www.genecards.org/cgi-bin/carddisp?MAP4K4&search=GCK-like+kinase&suff= txt this is a link to Weizman Institute's GeneCards (www.genecards.org) i entered GCK-like kinase and as best i can tell it believes that is the same as map kinase kinase kinase kinase 4? regardless, if that is not the right one, you might go ahead and try to find it on there using your own criteria. as you scroll down the page you will see expression as reported using any number of methodologies as well as links out with more information about the expression data. there is also a site called iHOP that has tons of information about proteins. dana ----- Original Message ----- From: "Edwards, R.E." To: Sent: Wednesday, January 25, 2006 9:29 AM Subject: [Histonet] RE: GCK-like kinase(GLK) --Anyone have any idea of the distribution of GLK in normal human tissues, unable to find anything in Pubmed. Thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER..U.K..... _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Wed, 25 Jan 2006 15:08:32 -0700 From: Gayle Callis Subject: Re: [Histonet] M2A To: "Patsy Ruegg" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20060125150737.01b4a340@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Patsy, Try this website, a new one for antibody location. www.exactantigen.com Good luck At 02:40 PM 1/25/2006, you wrote: >has anyone used an antibody M2A Antigen (Lymphatic Endothelial Cell Marker) Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 ------------------------------ Message: 16 Date: Wed, 25 Jan 2006 14:18:32 -0800 From: "Brusig, Stephanie" Subject: [Histonet] Sakura Tissue Tek II To: Message-ID: <16E971922EDF9A4B9E8D3216374EDD49766C30@wafedixm12.corp.weyer.pri> Content-Type: text/plain; charset="us-ascii" Does any one have a operators manual they don't need for a Tissue Tek II? I don't feel like paying the $125 from Sukura. Thanks, ~Stephanie Brusig Weyerhaeuser Company Propagation of High Value Trees WTC-1B10 253.924.6518 ------------------------------ Message: 17 Date: Wed, 25 Jan 2006 16:50:40 -0600 From: "Chaussey, Leslie" Subject: [Histonet] Positions available To: Message-ID: Content-Type: text/plain; charset="us-ascii" Northwestern Memorial Hospital, consistently recognized as the most preferred hospital in Chicago, is seeking new team members. We hire the 'Best People' who are dedicated to being a part of the 'Best Patient Experience!' At Northwestern Memorial Hospital, we're setting new standards of care. We're also creating career opportunities that allow you to share your talent, develop new skills, and reach all of your goals. We're one of the best hospitals in the world because we support the efforts, the ideas, and the initiatives of all of our people. We invite you to join us. We are currently seeking qualified candidates for our Histology, Gross Room, and Electron Microscopy/Renal Laboratories openings. For more information and to submit your resume, please visit our website, www.nmh.org . AA/EOE ----------------------------------------- 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. ------------------------------ Message: 18 Date: Wed, 25 Jan 2006 18:07:43 EST From: KDwyer3322@aol.com Subject: [Histonet] Texas Society for Histotechnology 2006 Program To: histonet@lists.utsouthwestern.edu Cc: , JWebb01@jpshealthnetwork.org, veronida@baylorhealth.edu Message-ID: <1a0.44b03f7c.31095ebf@aol.com> Content-Type: text/plain; charset="US-ASCII" To all: The Texas Society for Histotechnology will be having it's 2006 Symposium/Convention March 31-April 2, 2006 in Corpus Christi, Texas at the Omni Hotel. For a program or more information contact: Judy Webb - 817927-1024 or Veronica Davis -972-579-8291 Friday, March 31, 2006 10:00am-5:00 p.m. NSH Sponsored Workshop HT (ASCP) Examination Readiness - Glenda Hoye BS, HT (ASCP) Saturday, April 1, 2006 8-11:30 a.m. A.M. Workshops #1 Reagent Alcohol-Can't Drink It-So What is It? Pam Marcum HT(ASCP) #2 Technical Immunohistochemistry: Achieving Reliability and Reproducibility of Immunostains Rodney Miller, M.D. #3 Thinking LEAN in Histology R. Stephens HT(ASCP) QIHC #4. ASCP Certification Maintenance Program Evelyn Sandberg, HT(ASCP)HTL Saturday, April 1, 2006 1:00-4:30 p.m. P.M. Workshops #5 IHC Mathematics in the Laboratory Joel Martinez /Fatima Natar #6 MonKey Business-the Key to Delegation Jan Gardner ,MBA,HT(ASCP)/ Judi Stasko, BS, CLT #7 ISH/FISH Theory and Application Noemi Sebastiao #8. Gross Tissue Examination Charles Embrey,BS, HT(ASCP), PA(ASCP) Sunday,April 2, 2006 8:00-11:30 a.m. AM Workshops #9 CSI: Corpus Christi, Case Study Investigations Mike Reichenbach, HT (ASCP) QIHC Debra Flynn, HT(ASCP) QIHC #10 Ready or Not Here It Comes: Microwave Technology Donna Willis, HT(ASCP) HTL #11 Do You Know What Your Laboratory Workflow Is? Ritu Ward, MT(ASCP) MAOM #12. Making Job Descriptions Work for You Jan Gardner, MBA, HT (ASCP)/ Judi Stasko, BS, CLT ------------------------------ Message: 19 Date: Thu, 26 Jan 2006 09:07:06 -0000 From: "Nicola Cragg" Subject: [Histonet] Quantitative Image Analysis Systems - -Any recommendations? To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi All, Has anyone got any recommendations and/or advice on image analysis systems? We are looking to buy a new image analysis system to quantify IHC and perform histometric measurements (although this may require a second system). Does anyone have a system they can recommend? Or has anyone looked into this field and can offer any advice. Up to now, we've looked into Chromavision, Zeiss & Ariol. Thanks in advance, Nicola Cragg EpiStem Ltd. Manchester, UK ------------------------------ Message: 20 Date: Thu, 26 Jan 2006 06:00:40 -0500 From: "Bartlett, Jeanine" Subject: [Histonet] technique (Gimenez?) for rickettsiae To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Hi all: Can anyone recommend a good technique for demonstration of rickettsiae in FFPE tissue ? Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov ------------------------------ Message: 21 Date: Thu, 26 Jan 2006 07:30:30 -0500 From: "Mildred Fail" Subject: Re: [Histonet] technique (Gimenez?) for rickettsiae To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Jeanine, Do both a Giemsa and Himenez, rickettsiae is + in both Rena Fail Rena Fail >>> "Bartlett, Jeanine" 01/26/06 06:00AM >>> Hi all: Can anyone recommend a good technique for demonstration of rickettsiae in FFPE tissue ? Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 22 Date: Thu, 26 Jan 2006 08:05:11 -0500 From: "Bartlett, Jeanine" Subject: RE: [Histonet] technique (Gimenez?) for rickettsiae To: "Mildred Fail" , Message-ID: Content-Type: text/plain; charset="US-ASCII" They don't like the Giemsa for this: they want something that will make the bugs pop out at you. Jeanine Bartlett, HT(ASCP) (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: Mildred Fail [mailto:FAILM@musc.edu] Sent: Thursday, January 26, 2006 7:31 AM To: Bartlett, Jeanine; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] technique (Gimenez?) for rickettsiae Jeanine, Do both a Giemsa and Himenez, rickettsiae is + in both Rena Fail Rena Fail >>> "Bartlett, Jeanine" 01/26/06 06:00AM >>> Hi all: Can anyone recommend a good technique for demonstration of rickettsiae in FFPE tissue ? Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 23 Date: Thu, 26 Jan 2006 08:10:47 -0500 From: "Mildred Fail" Subject: [Histonet] CORRECTION To: Message-ID: Content-Type: text/plain; charset=US-ASCII gimenez and giemsa Rena Fail ------------------------------ Message: 24 Date: Thu, 26 Jan 2006 05:45:31 -0800 (PST) From: Kim Merriam Subject: Re: [Histonet] Quantitative Image Analysis Systems - -Any recommendations? To: Nicola Cragg , histonet@lists.utsouthwestern.edu Message-ID: <20060126134531.15492.qmail@web50306.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 We have an Aperio system (http://www.aperio.com/), it scans the entire slide at a 20X mag. The resolution is absolutely amazing, we compared all of the major brands mentioned below and this was by far the best system. It also comes with some nice image analysis software that can do pixel count, nuclear, membrane and they can be modified to fit your specific needs. We have the T2 system that can scan 120 slides at a time. It is definitely worth looking into. Kim Merriam Novartis Cambridge, MA Nicola Cragg wrote: Hi All, Has anyone got any recommendations and/or advice on image analysis systems? We are looking to buy a new image analysis system to quantify IHC and perform histometric measurements (although this may require a second system). Does anyone have a system they can recommend? Or has anyone looked into this field and can offer any advice. Up to now, we've looked into Chromavision, Zeiss & Ariol. Thanks in advance, Nicola Cragg EpiStem Ltd. Manchester, UK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. ------------------------------ Message: 25 Date: Thu, 26 Jan 2006 05:55:17 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] technique (Gimenez?) for rickettsiae To: "Bartlett, Jeanine" , histonet@lists.utsouthwestern.edu Message-ID: <20060126135517.46518.qmail@web61219.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Jeanine: Between 1911 and 1945 there were 21 methods/variants developed for rickettsiaeThe one by Wolbach (1919) stain nuclei and Rickettsiae blue and cytoplasm red. L?pine's (1932) stain Rickettsiae red in pale brown cells. Many others are variations of red agains blues (similar to Giemsa used for this purpose). I prefer Wolbach's. Hope this will help. Ren? J. "Bartlett, Jeanine" wrote: Hi all: Can anyone recommend a good technique for demonstration of rickettsiae in FFPE tissue ? Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. ------------------------------ Message: 26 Date: Thu, 26 Jan 2006 09:03:51 -0500 From: "Bartlett, Jeanine" Subject: RE: [Histonet] technique (Gimenez?) for rickettsiae To: "Rene J Buesa" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" We use the Wolbach's Giemsa but he doesn't care for it for this purpose. Thanks. Jeanine Bartlett, HT(ASCP) (404) 639-3590 jeanine.bartlett@cdc.hhs.gov _____ From: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: Thursday, January 26, 2006 8:55 AM To: Bartlett, Jeanine; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] technique (Gimenez?) for rickettsiae Jeanine: Between 1911 and 1945 there were 21 methods/variants developed for rickettsiaeThe one by Wolbach (1919) stain nuclei and Rickettsiae blue and cytoplasm red. L?pine's (1932) stain Rickettsiae red in pale brown cells. Many others are variations of red agains blues (similar to Giemsa used for this purpose). I prefer Wolbach's. Hope this will help. Ren? J. "Bartlett, Jeanine" wrote: Hi all: Can anyone recommend a good technique for demonstration of rickettsiae in FFPE tissue ? Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____ Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. ------------------------------ Message: 27 Date: Thu, 26 Jan 2006 10:01:30 -0500 From: "Boyce, Amanda \(NIH/NIAMS\) [F]" Subject: [Histonet] skeletal staining To: Message-ID: <72210BBE225D724FBDB0DF25C67B1A5305D99B@NIHCESMLBX9.nih.gov> Content-Type: text/plain; charset="iso-8859-1" I'm in the middle of doing alcian blue/alizarin red whole skeletal chick preps and I have a question. The protocol I've chosen uses 95% ethanol for the fixative. I've already fixed in PFA. Is it possible to continue with the protocol? Are there any steps I should add? Thanks, Amanda Boyce ------------------------------ Message: 28 Date: Thu, 26 Jan 2006 15:29:33 +0000 From: Ian Montgomery Subject: [Histonet] Advice. To: histonet@lists.utsouthwestern.edu Message-ID: <7.0.1.0.2.20060126151836.03b48eb0@bio.gla.ac.uk> Content-Type: text/plain; charset="us-ascii"; format=flowed With cuts in staff I've been asked if I can teach the histology portion of a zoology class. Problem, one of the tissue I'll be using is insect abdomen with its chitin exoskeleton. Any hints and tips regarding this type of material, processing, embedding, sectioning, that sort of thing. Or, is it Mollifix time again? Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk ------------------------------ Message: 29 Date: Thu, 26 Jan 2006 10:39:52 -0500 From: "Martha Ward" Subject: [Histonet] Hep Par 1 antibody To: Message-ID: <61135F0455D33347B5AAE209B903A3041209894B@EXCHVS2.medctr.ad.wfubmc.edu> Content-Type: text/plain; charset="us-ascii" I would like to thank everyone that responded to my question. I hope to get it up and running quickly. Martha Ward Wake Forest University Baptist Medical Center ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 26, Issue 31 **************************************** From ryaskovich <@t> dir.nidcr.nih.gov Thu Jan 26 12:17:50 2006 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR) [E]) Date: Thu Jan 26 12:18:00 2006 Subject: [Histonet] Advice. Message-ID: Gayle, That's from the Armed Forces Institute of Pathology Laboratory Methods in Histotechnology. Ruth Yaskovich National Institutes of Health National Institute of Dental and Crainiofacial Research Pain and Neurosensory Mechanism Branch Drug Discovery > ---------- > From: Gayle Callis > Sent: Thursday, January 26, 2006 12:55 PM > To: Ian Montgomery; Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Advice. > > For some reason, I have a chapter copied from a book titled Animal and > Insect Histotechnology, no author - all I have is a chapter from this book > authored by Debra A. McElroy - but it contained some words of wisdom for > insects. > > Fixation NBF, Bouins, or Zenkers - after fixation, soak fixed specimens in > 4% phenol (4 ml melted phenol in 96 ml 80% ethanol) for 24 hours to soften > the chitnous exoskeletaons then process into paraffin. > > If you try this, please keep us posted on results. > > > > At 08:29 AM 1/26/2006, you wrote: > > With cuts in staff I've been asked if I can teach the histology > > portion of a zoology class. Problem, one of the tissue I'll be using is > > insect abdomen with its chitin exoskeleton. Any hints and tips regarding > > this type of material, processing, embedding, sectioning, that sort of > > thing. Or, is it Mollifix time again? > > > >Dr. Ian Montgomery, > >Histotechnology, > >IBLS Support Services, > >Graham Kerr Building, > >Institute of Biomedical & Life Sciences, > >University of Glasgow, > >Glasgow, > >G12 8QQ. > >Tel: 0141 339 8855 > >Office: 4652 > >Lab: 6644. > >Pager: 07623 975451 > >e-mail: ian.montgomery@bio.gla.ac.uk > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From ana.merino-trigo <@t> wanadoo.fr Thu Jan 26 12:22:37 2006 From: ana.merino-trigo <@t> wanadoo.fr (ana.merino-trigo) Date: Thu Jan 26 12:23:04 2006 Subject: [Histonet] Paraffin cleaning product References: <0BE6ADFAE4E7E04496BF21ABD346628005653978@EXCHANGE1.huntingtonhospital.com> Message-ID: <016c01c622a5$7c229ff0$5b790a0a@BABA> Hi, I was wondering if anyone could suggest me a product to clean residues of paraffin in microtome, bench, tools... When working in Europe I was using a product "SafeSolv" from Labonord. In my actual lab at San Diego, they cannot order from this company and I'm not aware about products that I could find here. I search on the web "safesolv" and it comes several things, one is this link http://www.washwax.com/fc_purchase.html, could you please let me know if any of this products will do the job? I could use xylen to clean but microtome it's not on the hood to be using xylen and I don't have a easy access to a hood for this purpose. Thanks a lot, Ana From Malcolm.McCallum <@t> tamut.edu Thu Jan 26 12:39:01 2006 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Thu Jan 26 12:43:38 2006 Subject: [Histonet] tissue pics Message-ID: Hi everyone, Anyone out there have some nice digital images of diseased epithelium. Last week we covered various epithelium tissues, then had the bright idea that it might be cool to show some pathologies this week. Also, please tell me what it is in the email. I know some pathologies, but am not a pathologist, so I might not know what the particular condition is. I would just like to expose the students see some of this variation and have them try to figure out what it is! Thanks! Other tissues would also be welcomed for later in the semester. I hope this isn't to forward of a request! thanks if anyone can help! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Fred Underwood Sent: Thu 1/26/2006 9:54 AM To: Histonet@lists.utsouthwestern.edu; bill501@mindspring.com; Jeffery-Smith@ouhsc.edu; Terry.Marshall@rothgen.nhs.uk; kemlo.rogerson@waht.swest.nhs.uk Subject: RE: [Histonet] H.pylori immunos on gastric biopsies I would have bet $10 I wouldn't have seen that phrase today. >>> "Marshall Terry Dr, Consultant Histopathologist" 01/25/06 06:54AM >>> Yes, but a turd is easier to bottle. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:kemlo.rogerson@waht.swest.nhs.uk] Sent: 25 January 2006 09:02 To: Marshall Terry Dr, Consultant Histopathologist; Smith, Jeffery D. (HSC); Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies Isn't there a breath test too? Kemlo Rogerson Weston Super Mare Cell Path Manager -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, January 24, 2006 5:09 PM To: Smith, Jeffery D. (HSC); Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies H. pylori is a pathogen. It does not sit in the stomach behaving itself. Its presence is an indication for treatment, but its absence is *not* an indication for treatment directed against it, irrespective of how inflamed the stomach looks. The best histologic determinant of H. pylori is an immunostain. However, in most cases, this is all rather academic, as a quick stool test is available, directed against H. pylori antigen, so in most cases, we are really trying to determine something better done by other means. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Smith, Jeffery D. (HSC) [mailto:Jeffery-Smith@ouhsc.edu] Sent: 24 January 2006 16:18 To: Bill Blank; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies I have to somewhat agree. Immunos for H. pylori seems to be overkill. We routinely stain for H. pylori using a Giemsa on all GI biopsies. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bill Blank Sent: Tue 1/24/2006 9:29 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] H.pylori immunos on gastric biopsies IMO, we take this detecting H. pylori thing too seriously. A rational physician treats the patient and not any lab test. If I had symptoms of H. pylori, if my stomach looked inflamed, I would want to be treated for H. pylori irregardless of their presence on a biopsy. (Well, maybe as there is some evidence that treating antral H. pylori may increase the risk of GE junction adenocarcinoma) I consider immunos on gastric biopsies to be overkill and a waste of health care dollars. BIll At 7:53 AM -0600 1/24/06, GUTIERREZ, JUAN wrote: >We do immunos on all gastric biopsies. We had some cases that looked >negative by giemsa stain, yet the clinical hx suggested otherwise. When >we ran the immunos on these so called negatives, we were shocked to see >the amount of organisms we had missed. Do you want to take that chance? >I always ask myself: what if this was my biopsy? Something to think >about. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> cdc.gov Thu Jan 26 12:37:03 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Jan 26 12:53:45 2006 Subject: [Histonet] Paraffin cleaning product Message-ID: I like Para/Gard from TBS...I think VWR also sells it. Fisher has a similar product called Parapel, but I have not tried it. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 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 ana.merino-trigo Sent: Thursday, January 26, 2006 1:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin cleaning product Hi, I was wondering if anyone could suggest me a product to clean residues of paraffin in microtome, bench, tools... When working in Europe I was using a product "SafeSolv" from Labonord. In my actual lab at San Diego, they cannot order from this company and I'm not aware about products that I could find here. I search on the web "safesolv" and it comes several things, one is this link http://www.washwax.com/fc_purchase.html, could you please let me know if any of this products will do the job? I could use xylen to clean but microtome it's not on the hood to be using xylen and I don't have a easy access to a hood for this purpose. Thanks a lot, Ana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tracy.bergeron <@t> crl.com Thu Jan 26 12:59:12 2006 From: tracy.bergeron <@t> crl.com (tracy.bergeron@crl.com) Date: Thu Jan 26 12:59:30 2006 Subject: [Histonet] Paraffin cleaning product In-Reply-To: <016c01c622a5$7c229ff0$5b790a0a@BABA> Message-ID: Hi Ana, We use Para/Gard which is made by TBS. It is a great product and can be purchased through VWR. The link is below: http://www.vwrsp.com/catalog/product/index.cgi?catalog_number=15147-908&inE=1&highlight=15147-908 Tracy E. Bergeron, BS, HT (ASCP) Histotechnician Charles River Laboratories Wilmington, MA 978-658-6000 x 1229 "ana.merino-trigo" Sent by: histonet-bounces@lists.utsouthwestern.edu 01/26/2006 01:22 PM Please respond to "ana.merino-trigo" To cc Subject [Histonet] Paraffin cleaning product Hi, I was wondering if anyone could suggest me a product to clean residues of paraffin in microtome, bench, tools... When working in Europe I was using a product "SafeSolv" from Labonord. In my actual lab at San Diego, they cannot order from this company and I'm not aware about products that I could find here. I search on the web "safesolv" and it comes several things, one is this link http://www.washwax.com/fc_purchase.html, could you please let me know if any of this products will do the job? I could use xylen to clean but microtome it's not on the hood to be using xylen and I don't have a easy access to a hood for this purpose. Thanks a lot, Ana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LJohns53 <@t> CNTUS.JNJ.COM Thu Jan 26 14:14:41 2006 From: LJohns53 <@t> CNTUS.JNJ.COM (Johns, Laura [CNTUS]) Date: Thu Jan 26 14:14:54 2006 Subject: [Histonet] fixation of rat cerebellum Message-ID: <70F83FE9F65318468A612768E7043F8903754058@cntusmaexs9.na.jnj.com> Hi All, Does anyone have a suggestion for the length of time needed for fixation of rat cerebellum? We want to perform immunohistochemistry. Thanks, Laura Laura M. Johns, Ph.D. Centocor, Inc. 145 King of Prussia Road (Mail Stop # R-4-2) Radnor, PA 19087 Phone: 610-240-8284 .... Fax: 610-240-8150.... Email: ljohns53@cntus.jnj.com > Confidentiality Notice: This message is intended for the individual(s) 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, he/she is > 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 by telephone and delete > the e-mail from your system immediately. Thank you. > > From LJohns53 <@t> CNTUS.JNJ.COM Thu Jan 26 15:16:35 2006 From: LJohns53 <@t> CNTUS.JNJ.COM (Johns, Laura [CNTUS]) Date: Thu Jan 26 15:16:53 2006 Subject: [Histonet] fixation of rat cerebellum Message-ID: <70F83FE9F65318468A612768E7043F890375405A@cntusmaexs9.na.jnj.com> I am looking for astrocytes. Thanks, Laura -----Original Message----- From: Favara, Cynthia (NIH/NIAID) [E] [mailto:cfavara@niaid.nih.gov] Sent: Thursday, January 26, 2006 4:13 PM To: Johns, Laura [CNTUS] Subject: RE: [Histonet] fixation of rat cerebellum It may depend on what you are looking for. I routinely keep mouse/hamster brain for 7-10 days in NBF. We work with prions and do a number of IHC stains with no difficulties. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Johns, Laura [CNTUS] [mailto:LJohns53@CNTUS.JNJ.COM] Sent: Thursday, January 26, 2006 1:15 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] fixation of rat cerebellum Hi All, Does anyone have a suggestion for the length of time needed for fixation of rat cerebellum? We want to perform immunohistochemistry. Thanks, Laura Laura M. Johns, Ph.D. Centocor, Inc. 145 King of Prussia Road (Mail Stop # R-4-2) Radnor, PA 19087 Phone: 610-240-8284 .... Fax: 610-240-8150.... Email: ljohns53@cntus.jnj.com > Confidentiality Notice: This message is intended for the individual(s) 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, he/she is > 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 by telephone and delete > the e-mail from your system immediately. Thank you. > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vbaker60 <@t> yahoo.com Thu Jan 26 15:24:22 2006 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Thu Jan 26 15:24:33 2006 Subject: [Histonet] Nalgene wash tanks set inside carts (it's an off the wall question..sorry) Message-ID: <20060126212422.83922.qmail@web52509.mail.yahoo.com> Hi This question is aimed at facilities that have a central washroom and staff for cleaning and autoclaving. I'm in search of a cart that I can insert a 30gallon Nalgene wash tub into. A regular cart won't work because of the height (cart + tank)being too high. Where I work I have seen these tanks inserted into carts that fit the dimensions, but no one knows where they were purchased from. I've looked at Labconco, Nalgene, Rubbermaid and searched on the web without any luck. Does anyone have something like this. The dimensions of our tank is 34"L X 19"W X 36"H. I wish I could attach a picture but it's not possible. If anyone has something like this or any information I would really appreciate it. Thanks Vikki Baker Lab Manager The Godfrey Lab Mt. Sinai School of Medicine NYC, NY __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From RCazares <@t> schosp.org Thu Jan 26 15:26:28 2006 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Thu Jan 26 15:26:38 2006 Subject: [Histonet] Toenails and Nair Message-ID: <913FAC2B773C19488E26AE6572180FA50458A90D@exch01.schosp.org> Hello histonetters, I just wanted to share a tip I recently discovered. A tech that has just joined us on a full time basis, suggested leaving toenails in Nair overnight before processing. I had heard of using Nair before, but not leaving the nails in it overnight. Well, I tried it and the nails cut beautifully!! So I just had to share this with all of you out there in histoland. Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. From KevinMcGovern <@t> catholichealth.net Thu Jan 26 15:27:26 2006 From: KevinMcGovern <@t> catholichealth.net (McGovern, Kevin) Date: Thu Jan 26 15:27:41 2006 Subject: [Histonet] Paraffin cleaning product Message-ID: I don't know if they sell it in Europe, but Goo-Gone does a great job on paraffin, is easy to find, inexpensive, and smells like oranges! Kevin S. McGovern, BMETII Good Samaritan Hospital Clinical Engineering Dept. 10 East 31st Street Kearney, NE 68836 Phone: (308) 865-7051 Fax: (308) 865-2914 e-Mail: kevinmcgovern@catholichealth.net Confidentiality Notice: This email, including any attachments, contains CONFIDENTIAL information. The information is intended only for the use of the individual(s) or entity name above. If you are not the intended recipient, you are notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this email is not permissible. If you have received this email in error, please notify the sender immediately by reply mail, and destroy all copies of the original message. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bartlett, Jeanine Sent: Thursday, January 26, 2006 12:37 PM To: ana.merino-trigo; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin cleaning product I like Para/Gard from TBS...I think VWR also sells it. Fisher has a similar product called Parapel, but I have not tried it. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 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 ana.merino-trigo Sent: Thursday, January 26, 2006 1:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin cleaning product Hi, I was wondering if anyone could suggest me a product to clean residues of paraffin in microtome, bench, tools... When working in Europe I was using a product "SafeSolv" from Labonord. In my actual lab at San Diego, they cannot order from this company and I'm not aware about products that I could find here. I search on the web "safesolv" and it comes several things, one is this link http://www.washwax.com/fc_purchase.html, could you please let me know if any of this products will do the job? I could use xylen to clean but microtome it's not on the hood to be using xylen and I don't have a easy access to a hood for this purpose. Thanks a lot, Ana _______________________________________________ Histonet mailing list Histonet@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 glorialimetti <@t> yahoo.com Thu Jan 26 16:47:09 2006 From: glorialimetti <@t> yahoo.com (Gloria Limetti) Date: Thu Jan 26 16:47:19 2006 Subject: [Histonet] Re: Histonet Digest, Vol 26, Issue 19 Message-ID: <20060126224709.32917.qmail@web52407.mail.yahoo.com> re:paraformaldehyde In our lab we use the para powder and yes it is very airborne. But....we carefully weigh out the powder with the hood sash completly up or using a mask outside of the hood. The we carefully place it in a ziplock bag. Whe we need it we just cut the end and slowly poor it into our water. This eliminates alot of powder in the air ...just a thought... Gloria Limetti University of Pgh Vestibular Research --- histonet-request@lists.utsouthwestern.edu wrote: > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, > visit > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body > 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it > is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. Re: Dry Stains Shelf Life (John A. Kiernan) > 2. Re: paraformaldehyde crystals v Prills (John > A. Kiernan) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Tue, 17 Jan 2006 12:41:28 -0500 > From: "John A. Kiernan" > Subject: Re: [Histonet] Dry Stains Shelf Life > To: Sharon E Willman > Cc: histonet@lists.utsouthwestern.edu > Message-ID: <43CD2C48.F22D41E4@uwo.ca> > Content-Type: text/plain; charset=us-ascii > > See the following publications: > > Emmel VM & Stotz EH 1986. Certified biological > stains: a stability study. Stain Technol. > 61:385-387. > Titford M 2001. Comparison of historic Grubler > dyes with modern counterparts. Biotech. Histochem. > 76:23-30. > Titford, M. 2002. Save that dye! Microscopy Today > 18-5:31-34. > > The general conclusion is that nearly all dye > powders still work after 100 years (Titford). > Quantitative assays show very little deterioration > over 20 or so years (Emmel & Stotz). Matt Frank > may have more recent information from the > Biological Stain Commission's lab. > > Alcian blue 8G can deteriorate in the solid state, > changing to an insoluble pigment. This is not a > problem with the pyridine variant of alcian blue > (Churukian et al 2000, Biotech. Histochem. 75: > 147-150). Acidic solutions of alcian blue 8G are > stable for some years on my shelves. Churukian's > lab manual gives a recommended shelf life of 6 > months. An alcian blue solution with a precipitate > should be discarded and replaced, not filtered and > used. > -- > ------------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > http://instruct.uwo.ca/anatomy/530/index.htm > _______________________________ > Sharon E Willman wrote: > > > > Hi, > > What is the expiration time for dry stains and how > often should you > > replenish? Many times they do not have expiration > dates on them when > > they come in from the manufacturer. > > I would appreciate any information on this matter. > > Thanks, > > Sharon Willman > > > > > > ------------------------------ > > Message: 2 > Date: Tue, 17 Jan 2006 12:55:20 -0500 > From: "John A. Kiernan" > Subject: Re: [Histonet] paraformaldehyde crystals v > Prills > To: Carl Hobbs > Cc: histonet@lists.utsouthwestern.edu > Message-ID: <43CD2F88.7CA05654@uwo.ca> > Content-Type: text/plain; charset=us-ascii > > Carl Hobbs wrote: > > > > Many of our Groups use paraformaldehyde crystals ( > Sigma P6148) to make up > > their solution of formalin. Works very well; 4g + > 1tab PBS + 6microlitre > > 10MNaOH.....stir overnight. Dissolved > completely:pH7.2 > > I am trying to get away from using this > paraformaldehyde: it is very light > > and prone to spread around the fume hood when it's > weighed out. > > So, I bought PRILLS from Sigma:Problem is, > they just do not dissolve > > completely, unless I preboil the water.( heating > to 60C is not hot enough to > > dissolve the Prills. > > I have tried all combinations of the formulation > above( eg: no NaOH, > > dissolving PBS tablet before adding > paraformaldehyde). > > I want the procedure with the least risk; boiling > may be a higher risk than > > the spread of powder. > > NB: I cannot turn down/off the fume hoods, for the > weighing of the > > crystals( which behave more like a very fine > powder), hence my desire to use > > the heavier Prills. > > > > Be grateful for any insights. > > Carl > > You can avoid the fine powder and insolubility > problems by making your fixative from formalin > rather than paraformaldehyde. > > A good mixture, shown to be OK even for electron > microscopy, is that of Carson FL, Martin JH & Lynn > JA 1973, Am. J. Clin. Path. 59:365-373. Its > composition is: > Formalin 100 ml > Water 900 ml > Monobasic sodium phosphate, monohydrate 18.6 g > Sodium hydroxide 4.2 g > The pH is 7.2-7.4 and the solution can be kept for > several months. > > As always, don't rely on the accuracy of this or > any other email! Consult the original paper before > making and using the fixative. > -- > ------------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > http://instruct.uwo.ca/anatomy/530/index.htm > _______________________________ > > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 26, Issue 19 > **************************************** > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From glorialimetti <@t> yahoo.com Thu Jan 26 16:48:57 2006 From: glorialimetti <@t> yahoo.com (Gloria Limetti) Date: Thu Jan 26 16:49:06 2006 Subject: [Histonet] Re: Histonet Digest, Vol 26, Issue 14 Message-ID: <20060126224857.15227.qmail@web52413.mail.yahoo.com> RE; Eosin too pink! We love the pink! but just dilute it woth more 95% or stain for less time. Gloria Limetti University of Pittsburgh/Vestibular Research --- histonet-request@lists.utsouthwestern.edu wrote: > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, > visit > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body > 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it > is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. RE: Eosin too pink > (Marshall Terry Dr, Consultant > Histopathologist) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Fri, 13 Jan 2006 17:54:18 -0000 > From: "Marshall Terry Dr, Consultant > Histopathologist" > > Subject: RE: [Histonet] Eosin too pink > To: "Parker, Helayne" , > > Message-ID: > > > Content-Type: text/plain; charset="iso-8859-1" > > Wouldn't we all *love* to see just what is > considered too pink:-) > An old pathologist friend of mine used to put a > pinch of bicarb in the eosin when the techs were not > looking. > He had a thing of not liking it too pink. For me, I > can't have it too pink, as long as the nuclei are > blue. > > Dr Terry L Marshall, B.A.(Law), > M.B.,Ch.B.,F.R.C.Path > Consultant Pathologist > Rotherham General Hospital > South Yorkshire > England > terry.marshall@rothgen.nhs.uk > > -----Original Message----- > From: Parker, Helayne [mailto:HParker@Skaggs.Net] > Sent: 13 January 2006 16:42 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Eosin too pink > > > Hi List, > We are currently doing our H & E stain by hand and > using the > Richard-Allan 7211 Hematoxylin and their Eosin Y. > Our pathologist is > still complaining of slides being TOO PINK. I have > cut the eosin step > down to just 5 dips from 15 dips, and still too > pink. The Eosin is very > rich. > > This is our protocol: > Hydrate to water like normal (thru Clear Rite 3, > 100% and 95% Flex) > Hematoxylin- 2 mins > Rinsing in water until the water runs clear > Clarifier II- 30 dips > running water -30 dips > Richard Allan Bluing - 30 dips > running water - 30 dips > 95% Flex - 10 dips > Eosin-Y- 5 quick dips > Thru 95%, 100% etc to xylene > > Any suggestions on a possible method change or a > more mild Eosin. > Complaint seems to solely "TOO PINK". I have done > some experimenting w/ > cutting the eosin w/ 80% Flex- and it does calm it > down a bit but, the > Hemo looks more purply then blue, almost periwinkle > so not sure if that > will be better or not- we still need to have him > review the test slides. > Any suggestions ? > > Helayne Parker > Histology Section Head > Skaggs Community Health Center > Branson, Missouri > > > _______________________________________________ > Histonet mailing list > Histonet@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 26, Issue 14 > **************************************** > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From maria <@t> ski.org Thu Jan 26 18:38:06 2006 From: maria <@t> ski.org (Maria Mejia) Date: Thu Jan 26 18:38:31 2006 Subject: [Histonet] Advice. References: <7.0.1.0.2.20060126151836.03b48eb0@bio.gla.ac.uk> Message-ID: <43D96B6E.8070909@ski.org> Ian, Hmmm...well, some years back I was working on a butterfly heads project for a brain cell count study. Never worked on insects before, so it didn't take long before I noticed the head of the butterfly is covered with chitin layers and I had to section the entire head. Here's what I did. I got beautiful paraffin sections stained w/Nissl staining. With agitation: > fixed cut head in Carnoy's fixative - overnight @ 4C (there are many formulas in the literature. Possible could also use glutaraldehyde containing fixative - sure these would be fine for LM too) > dehydrate a bit more with ethanol alcohol 2x - 30' each and then > soak (tissue) in a mixture of equal parts of chloral hydrate:phenol to soften the entire chitin. *Chloral hydrate:phenol made by weighing equal parts & then gently warming until molten. Once molten can store for several weeks. Sections cut beautiful. Tissues were left in this mixture for a minimum of 2 hours to overnight (perhaps even up to a couple days). > clear in chloroform 3x - over 2 hours to overnight (I did 1 hour each). > place in cedarwood oil - 2 days (change oil 3x during period). > then do routine paraffin. *When sectioning, I remember adding a bit of glycerin in the ice water to soak the blocks a bit. Ian, I hope this helps. Maria Bartola Mejia Smith-Kettewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 Ian Montgomery wrote: > With cuts in staff I've been asked if I can teach the > histology portion of a zoology class. Problem, one of the tissue I'll > be using is insect abdomen with its chitin exoskeleton. Any hints and > tips regarding this type of material, processing, embedding, > sectioning, that sort of thing. Or, is it Mollifix time again? > > Dr. Ian Montgomery, > Histotechnology, > IBLS Support Services, > Graham Kerr Building, > Institute of Biomedical & Life Sciences, > University of Glasgow, > Glasgow, > G12 8QQ. > Tel: 0141 339 8855 > Office: 4652 > Lab: 6644. > Pager: 07623 975451 > e-mail: ian.montgomery@bio.gla.ac.uk > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jlinda <@t> ces.clemson.edu Thu Jan 26 19:13:48 2006 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Thu Jan 26 19:10:41 2006 Subject: [Histonet] Region III Hotel Deadline Message-ID: <5.2.1.1.2.20060126200430.00a8adb8@mailhost.ces.clemson.edu> Friday, January 27th is the cut-off date for hotel reservations at the Charleston Harbor Resort and Marina, the official hotel for the Region III meeting being held March 10 -12. Many of you may have had experience with planning a meeting and know that room reservations help defray the cost of a meeting. If you have made a reservation at another hotel in the area, please contact me at 864-238-8230. Have I got a deal for you! The phone number for the hotel is 843-856-0028 or 888-485-0136. Ask for the special rate for the Region III meeting. Charleston Awaits! Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo.htm From FARFAT <@t> aol.com Thu Jan 26 22:17:08 2006 From: FARFAT <@t> aol.com (FARFAT@aol.com) Date: Thu Jan 26 22:17:21 2006 Subject: [Histonet] bone marrow stain on smears Message-ID: <204.110af4c1.310af8c4@aol.com> Hi, I need some help in staining bone marrow smears fixed in methnol.I tried Giemsa and May grenwal stains but the pathologist are looking for better quality staining,if anybody has success in doing a giemsa stain on bone marrow smears and would like to share the technique with the name of the manufacture who makes the stain, it will be greatly appreciated. Thanks. Akbar. From martasantos <@t> ecsaude.uminho.pt Fri Jan 27 04:46:51 2006 From: martasantos <@t> ecsaude.uminho.pt (Marta Santos) Date: Fri Jan 27 04:35:03 2006 Subject: [Histonet] Rat MCL2a and MLC1a antibodies Message-ID: Dear all Does anyone has any experience with antibodies agaist rat myosin light chain 1a and 2a? I was not able to find any company with these products. Thanks in advance for all your help. Best Regards Marta Marta Santos Instituto de Ci?ncias da Vida e da Sa?de Escola de Ci?ncias da Sa?de Campus de Gualtar 4710-057 Braga Portugal E-mail : martasantos@ecsaude.uminho.pt From Lobke.DeBels <@t> UGent.be Fri Jan 27 05:32:42 2006 From: Lobke.DeBels <@t> UGent.be (Lobke De Bels) Date: Fri Jan 27 05:32:58 2006 Subject: [Histonet] celloidin Message-ID: <1138361562.43da04da27efb@mail.ugent.be> Dear all, We wanna try celloidin embedding for whole eyeballs but we don't have any experience with that technique. Could someone give us some information and/or protocols to start the celloidin embedding technique? Many thanks in advance for any help. With the kindest regards, Lobke De Bels Department of Morphology Faculty of Veterinary Medicine Ghent University Salisburylaan 133 9820 Merelbeke Belgium lobke.debels@ugent.be -- From mohana_g2002 <@t> yahoo.com Fri Jan 27 07:25:58 2006 From: mohana_g2002 <@t> yahoo.com (MOHANA) Date: Fri Jan 27 07:26:12 2006 Subject: [Histonet] cresyl violet staining Message-ID: <20060127132558.60734.qmail@web33504.mail.mud.yahoo.com> i was wondering if any one can explain me that why does the cresyl violet stains only neurons and not glia???? mohana Mohana --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. From kweidenh <@t> montefiore.org Fri Jan 27 08:08:07 2006 From: kweidenh <@t> montefiore.org (Karen Weidenheim) Date: Fri Jan 27 08:10:57 2006 Subject: [Histonet] cresyl violet staining Message-ID: Mohana, the cresyl violet dye has a great affinity for the Nissl substance (rough endoplasmic reticulum) in the neurons, so therefore it is used to highlight the neurons. However, other cell types (the various glia) also have some endoplasmic reticulum and they will stain at least somewhat. The reason for using thicker sections for the stain is so the laminar pattern of the neurons in aggregate can be seen, and any abnormalities found. So the low power view is important. So then the pathologist has to interpret the stain with some intelligence because of this, and may require additional staining to help them, depending on whether the case is a tumor or an epilepsy or something else. Researchers often have to be reminded that the stained cells are not exclusively neurons and that they need to interpret carefully. The immunostains for neurons including NeuN and neurofilament can supplement the Nissl (cresyl violet) method and glial staining i.e. GFAP may also help. Hope this helps. Karen Karen M. Weidenheim, M.D. Professor of Pathology, Clinical Neurology and Clinical Neurosurgery Albert Einstein College of Medicine Chief, Division of Neuropathology Montefiore Medical Center 111 East 210th Street Bronx, NY 10467 (718) 920-4446 FAX (718) 653-3409 Beeper (917) 556 3696 CONFIDENTIALITY NOTICE: This email, including any attachments, is for the sole use of the intended recipient(s). The information contained in this message may be private and confidential, and may also be subject to the work product doctrine. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Thank you. >>> MOHANA 01/27/06 8:25 AM >>> i was wondering if any one can explain me that why does the cresyl violet stains only neurons and not glia???? mohana Mohana --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mohana_g2002 <@t> yahoo.com Fri Jan 27 08:22:59 2006 From: mohana_g2002 <@t> yahoo.com (MOHANA) Date: Fri Jan 27 08:23:09 2006 Subject: [Histonet] cresyl violet Message-ID: <20060127142259.44618.qmail@web33509.mail.mud.yahoo.com> i have used cresyl violet with luxol fast blue stain for stainig hippocampal neurons in chick 12 micron thickness. but, i was just wondering as some glia would also be stained so how do i differenciate between the two ????? mohana Mohana --------------------------------- Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. From rjbuesa <@t> yahoo.com Fri Jan 27 09:55:16 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Jan 27 09:55:27 2006 Subject: [Histonet] bone marrow stain on smears In-Reply-To: <204.110af4c1.310af8c4@aol.com> Message-ID: <20060127155516.97572.qmail@web61220.mail.yahoo.com> Akbar: We used regular stains for BM, but all the dilutions were done with phosphate buffer at pH7, otherwise results were unsatisfactory. Hope this will help you. Ren? J. FARFAT@aol.com wrote: Hi, I need some help in staining bone marrow smears fixed in methnol.I tried Giemsa and May grenwal stains but the pathologist are looking for better quality staining,if anybody has success in doing a giemsa stain on bone marrow smears and would like to share the technique with the name of the manufacture who makes the stain, it will be greatly appreciated. Thanks. Akbar. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Now share photos without attaching a thing. Check out PhotoMail from Yahoo! Mail. From Mara.Fairman <@t> RoperSaintFrancis.com Fri Jan 27 10:51:31 2006 From: Mara.Fairman <@t> RoperSaintFrancis.com (Fairman Mara) Date: Fri Jan 27 10:51:49 2006 Subject: [Histonet] 5 GALLON PLASTIC XYLENE CONTAINERS Message-ID: I have a question for anyone ordering xylene or toluene in the 5 gallon heavy white plastic containers. The cost for disposal companies to take away is high. The landfills won't take unless they are: triple rinsed, all warning labels removed and large holes in them (so they can't be re-used). It is not an easy task to remove those labels, not to mention finding a way to put holes in that heavy plastic. I would appreciate hearing how other labs are disposing of these. Thanks, Mara Our Mission: Healing all people with compassion, faith and excellence Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain confidential and/or legally privileged information. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Any unauthorized review, use, disclosure, or distribution is prohibited. Thank you. From schnegelsberg <@t> xgene.com Fri Jan 27 12:29:55 2006 From: schnegelsberg <@t> xgene.com (Birthe Schnegelsberg) Date: Fri Jan 27 12:29:57 2006 Subject: [Histonet] in vitro skin Fb staining w/vimentin In-Reply-To: <20060124164828.11569.qmail@web61214.mail.yahoo.com> Message-ID: Hello. I am staining cells in in vitro skin with the fibroblast specific marker vimentin to differentiate between fibroblasts and keratinocytes. I am able to detect fluorescent signals in the tissue. The staining pattern and the cell localization makes me wonder though, if the staining is in fact specific fibroblast staining or if the antibody (immunon) picks up dead cell material. Does anybody have experienece using vimentin as a marker for fibroblast cells? The staining pattern of the positive control looks different than that for the singel cells in the in vitro skin?! Thanks, Birthe From Luis.Chiriboga <@t> med.nyu.edu Fri Jan 27 12:17:03 2006 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Jan 27 12:45:15 2006 Subject: [Histonet] NYSHS Website Message-ID: Hi Everyone The New York State Histotechnology Society has just opened it's new website. This year, NYSHS is hosting the Region 1 Symposium and some preliminary information is available on the site. To visit the site just click on the following link: www.nyhisto.org Have a great weekend.... Luis ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 From chengkaz <@t> uci.edu Fri Jan 27 13:17:06 2006 From: chengkaz <@t> uci.edu (Chengkang ZHANG) Date: Fri Jan 27 13:17:16 2006 Subject: [Histonet] Re: cresyl violet staining vs. Haematoxylin staining Message-ID: <43DA71B2.6080307@uci.edu> I am currently using haematoxylin to stain nuclei of neurons, but someone told me that cresyl violet is better than haematoxylin. Would anyone tell me what the difference between those two staining and which one you prefer for nuclei staining? Thanks Chengkang -- =================================== Chengkang Zhang Ph.D. Department of Pharmacology University of California, Irvine Irvine, CA 92697-4625 Email: chengkaz@uci.edu TEL: (949)-824-1902 (lab) FAX: (949)-824-4855 =================================== From ajohnson <@t> aipathology.com Fri Jan 27 13:38:40 2006 From: ajohnson <@t> aipathology.com (Amy Johnson) Date: Fri Jan 27 13:45:54 2006 Subject: [Histonet] FW: See if you can figure it out!!!!! Message-ID: <016A64931E1ED511B12C0002B3026080523538@SERV001> _____ From: Vercimak, JoAnn [mailto:jvercimak@graebel.com] Sent: Friday, January 27, 2006 8:02 AM To: Amy Johnson; Nordquist, Sherry; NorrbFarms@aol.com; lbelanger@eastbay.com; hbowers2@eastbay.com Subject: FW: See if you can figure it out!!!!! -----Original Message----- From: Rolain, Stacy Sent: Friday, January 27, 2006 7:54 AM To: Thorsheim, Todd; Cynkar, Meganne; Vercimak, JoAnn; Beeber, Rosie; Thronson, Michelle Subject: FW: See if you can figure it out!!!!! Stacy Rolain Graebel Lightning/Phoenix 930 Graebel San Antonio 956 General Ledger Accountant Work hours: 7:00 am to 4:00 pm Phone: 715-848-6605 Fax: 715-848-6334 I G R A E B E L |||'""|"""\___ I_______________|)| | | |_|o] (@)(@)"""""""""(@)(@)***(@) _____ From: Edwards, Eva Sent: Fri 1/27/2006 6:10 AM To: Braker, Bobbie; Brisby, Amy; Ingram, Ashley; Bliese, Amber; Nelson, Britta Subject: FW: See if you can figure it out!!!!! Eva Edwards NonInterstate, HHG-Intra-Local Revenue Billing & Pricing Specialist Graebel Companies 902, 918, 921 Hours: M-TH 7am-4pm, F 7am-3:30pm 715-848-6760 phone 715-848-6591 fax evaedwards@graebel.com I G R A E B E L |||'""|"""\___ I_______________|)| | | |_|o] (@)(@)"""""""""(@)(@)***(@) _____ From: EVA EDWARDS [mailto:evae44@msn.com] Sent: Thu 1/26/2006 3:57 PM To: Edwards, Eva Cc: letr0915@yahoo.com; james.solin@us.army.mil; 1thumper_1@charter.net; johnslilladie@yahoo.com; lisabobby92@yahoo.com; rodemmer@hotmail.com Subject: Fw: See if you can figure it out!!!!! >From: "kelly ison" >To: >Subject: Fw: See if you can figure it out!!!!! >Date: Thu, 26 Jan 2006 11:50:47 -0600 > > > > > > Subject: Fw: See if you can figure it out!!!!! > > A RIDDLE THAT'LL KILL YOUR BRAIN! > > This is going to make you so MAD! There are three words in the >English language that end in "gry". ONE is angry and the other is hungry. >EveryONE knows what the third ONE means and what it stands for. EveryONE >uses them everyday, and if you listened very carefully, I've given you the >third word. What is it? _______gry? Send this to 5 People and the answer >will pop up on the screen automatically. > > > > > > > > > > > > . > > > > From ajohnson <@t> aipathology.com Fri Jan 27 14:17:24 2006 From: ajohnson <@t> aipathology.com (Amy Johnson) Date: Fri Jan 27 14:24:16 2006 Subject: [Histonet] My Apologies Message-ID: <016A64931E1ED511B12C0002B302608052353A@SERV001> Dear Histonetters, I am so sorry for my previous email. I hit the histonet link instead of the person I really wanted to send to. So sorry for the inconvenience. From bjdewe <@t> aol.com Fri Jan 27 14:31:28 2006 From: bjdewe <@t> aol.com (bjdewe@aol.com) Date: Fri Jan 27 14:31:49 2006 Subject: [Histonet] FW: See if you can figure it out!!!!! In-Reply-To: <016A64931E1ED511B12C0002B3026080523538@SERV001> References: <016A64931E1ED511B12C0002B3026080523538@SERV001> Message-ID: <8C7F197C8FFC857-C18-5B3@MBLK-R08.sysops.aol.com> >> "Angry" and "hungry" are two words that end in "gry". There are three words in the English language. What is the third word? Everyone knows what it means and everyone uses it every day. Look closely and I have already given you the third word. What is it? Answer: "language". This puzzle has circulated widely on the Internet for some years, but usually in an abbreviated form such as "Name three common English words ending in 'gry'", which has no good third answer. http://www.word-detective.com/gry.html. ******************************************************************* 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. ******************************************************************* -----Original Message----- From: Amy Johnson To: Christine Johnson (tcjohnson_01@earthlink.net) ; Dad (poock@uniontel.net) ; Histonet (histonet@lists.utsouthwestern.edu) ; Laurie Johnson (laurie@plainfieldtrucking.com) ; Naomi Poock (naomip@thephoenixgrp.net) Sent: Fri, 27 Jan 2006 13:38:40 -0600 Subject: [Histonet] FW: See if you can figure it out!!!!! _____ From: Vercimak, JoAnn [mailto:jvercimak@graebel.com] Sent: Friday, January 27, 2006 8:02 AM To: Amy Johnson; Nordquist, Sherry; NorrbFarms@aol.com; lbelanger@eastbay.com; hbowers2@eastbay.com Subject: FW: See if you can figure it out!!!!! -----Original Message----- From: Rolain, Stacy Sent: Friday, January 27, 2006 7:54 AM To: Thorsheim, Todd; Cynkar, Meganne; Vercimak, JoAnn; Beeber, Rosie; Thronson, Michelle Subject: FW: See if you can figure it out!!!!! Stacy Rolain Graebel Lightning/Phoenix 930 Graebel San Antonio 956 General Ledger Accountant Work hours: 7:00 am to 4:00 pm Phone: 715-848-6605 Fax: 715-848-6334 I G R A E B E L |||'""|"""\___ I_______________|)| | | |_|o] (@)(@)"""""""""(@)(@)***(@) _____ From: Edwards, Eva Sent: Fri 1/27/2006 6:10 AM To: Braker, Bobbie; Brisby, Amy; Ingram, Ashley; Bliese, Amber; Nelson, Britta Subject: FW: See if you can figure it out!!!!! Eva Edwards NonInterstate, HHG-Intra-Local Revenue Billing & Pricing Specialist Graebel Companies 902, 918, 921 Hours: M-TH 7am-4pm, F 7am-3:30pm 715-848-6760 phone 715-848-6591 fax evaedwards@graebel.com I G R A E B E L |||'""|"""\___ I_______________|)| | | |_|o] (@)(@)"""""""""(@)(@)***(@) _____ From: EVA EDWARDS [mailto:evae44@msn.com] Sent: Thu 1/26/2006 3:57 PM To: Edwards, Eva Cc: letr0915@yahoo.com; james.solin@us.army.mil; 1thumper_1@charter.net; johnslilladie@yahoo.com; lisabobby92@yahoo.com; rodemmer@hotmail.com Subject: Fw: See if you can figure it out!!!!! >From: "kelly ison" >To: >Subject: Fw: See if you can figure it out!!!!! >Date: Thu, 26 Jan 2006 11:50:47 -0600 > > > > > > Subject: Fw: See if you can figure it out!!!!! > > A RIDDLE THAT'LL KILL YOUR BRAIN! > > This is going to make you so MAD! There are three words in the >English language that end in "gry". ONE is angry and the other is hungry. >EveryONE knows what the third ONE means and what it stands for. EveryONE >uses them everyday, and if you listened very carefully, I've given you the >third word. What is it? _______gry? Send this to 5 People and the answer >will pop up on the screen automatically. > > > > > > > > > > > > . > > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Fri Jan 27 15:26:03 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Jan 27 15:26:22 2006 Subject: [Histonet] 5 GALLON PLASTIC XYLENE CONTAINERS Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717659@lsexch.lsmaster.lifespan.org> We buy xylene in 1-gallon plastic bottles, and there is no problem putting them in the trash. However, just a couple of suggestions - Do you actually have to remove the labels, or will they accept painting them over? Or sanding them with coarse sandpaper so they are illegible? Probably the easiest way to make holes in such a container would be a hole saw set in an electric drill. I would do this after the containers are well rinsed. Electrical motors shouldn't be used in close proximity to flammable solvents for obvious reasons. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Fairman Mara > Sent: Friday, January 27, 2006 8:51 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] 5 GALLON PLASTIC XYLENE CONTAINERS > > I have a question for anyone ordering xylene or toluene in the 5 gallon > heavy white plastic containers. The cost for disposal companies to take > away is high. The landfills won't take unless they are: triple rinsed, > all > warning labels removed and large holes in them (so they can't be re-used). > It is not an easy task to remove those labels, not to mention finding a > way > to put holes in that heavy plastic. I would appreciate hearing how other > labs are disposing of these. > Thanks, > Mara > > > > > > > Our Mission: > Healing all people with compassion, faith and excellence > > Confidentiality Notice: > This e-mail message, including any attachments, is for > the sole use of the intended recipients and may contain > confidential and/or legally privileged information. > If you are not the intended recipient, please contact > the sender by reply e-mail and destroy all copies of the > original message. Any unauthorized review, use, > disclosure, or distribution is prohibited. > Thank you. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From sluhisto <@t> yahoo.com Fri Jan 27 16:13:27 2006 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Jan 27 16:13:36 2006 Subject: [Histonet] Gyn cytologies Message-ID: <20060127221327.13425.qmail@web51011.mail.yahoo.com> Hello All: Our lab may be getting into the gyn cytology business staining pap smears. It has been many, many, many years since I have done this. I'm sure that there are many advances. If any of you are doing gyn cytologies in your labs, would you mind sharing your expertise with me? Are you manual or is there automation for staining pap smears? What companies do you get your staining reagents from, etc? Thanks so much. You are all always a wealth of info. Have a super weekend!! Susan --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From LINDA.MARGRAF <@t> childrens.com Fri Jan 27 16:50:18 2006 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Fri Jan 27 16:50:42 2006 Subject: [Histonet] Histonet's 10th anniversary! Message-ID: Dear Histonet members: I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! There are currently 2342 members on the list from more than 30 different countries. I have no guess how many messages have gone through the server during this time. Must be millions. I'd like to personally thank all the members who actively participate in the list, making it such a worthwhile endeavor. Thanks so much. Thanks too to Herb for his behind the scenes computer expertise. Sincerely, Linda M Histonet administrator From jhabecke <@t> fhcrc.org Fri Jan 27 17:21:44 2006 From: jhabecke <@t> fhcrc.org (Randolph-Habecker, Julie) Date: Fri Jan 27 17:21:53 2006 Subject: [Histonet] Stains for Hemaglobin Message-ID: <21F1955FA284134E981948D6D3FFD04202ED3736@harpo.fhcrc.org> I am looking for a specific stain for hemoglobin in formalin-fixed, paraffin-embedded canine tissue. I could either use IHC or a special stain. Any ideas? I found a reference for a Puchtler-Rosenthal-Sweat Method for hemoglobin but it says that it only works in tissue fixed in Zenker's. Just wondering if anyone has any experience in this before we try it. Thanks, Julie Julie Randolph-Habecker, Ph.D. Staff Scientist - Manager Experimental Histopathology Shared Resource Fred Hutchinson Cancer Research Center 1100 Fairview Ave, N. M5-A803 Seattle WA 98109-1024 206-667-6119 jhabecke@fhcrc.org From Luis.Chiriboga <@t> med.nyu.edu Fri Jan 27 17:41:37 2006 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Jan 27 17:40:13 2006 Subject: [Histonet] Histonet's 10th anniversary! In-Reply-To: Message-ID: Don't thank us, THANK YOU !!! !!! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of LINDA MARGRAF Sent: Friday, January 27, 2006 5:50 PM To: Histonet@lists.utsouthwestern.edu Cc: hagler.herb@pathology.swmed.edu Subject: [Histonet] Histonet's 10th anniversary! Dear Histonet members: I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! There are currently 2342 members on the list from more than 30 different countries. I have no guess how many messages have gone through the server during this time. Must be millions. I'd like to personally thank all the members who actively participate in the list, making it such a worthwhile endeavor. Thanks so much. Thanks too to Herb for his behind the scenes computer expertise. Sincerely, Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dellav <@t> musc.edu Fri Jan 27 17:49:04 2006 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Jan 27 17:49:37 2006 Subject: [Histonet] Histonet's 10th anniversary! Message-ID: Congratulations Histonet and to all of you who make this a great resource and a wonderful community of dedicated professionals! Vinnie Della Speranza President, National Society for Histotechnology >>> "LINDA MARGRAF" 01/27/06 05:50PM >>> Dear Histonet members: I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! There are currently 2342 members on the list from more than 30 different countries. I have no guess how many messages have gone through the server during this time. Must be millions. I'd like to personally thank all the members who actively participate in the list, making it such a worthwhile endeavor. Thanks so much. Thanks too to Herb for his behind the scenes computer expertise. Sincerely, Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dellav <@t> musc.edu Fri Jan 27 17:54:08 2006 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Jan 27 17:54:34 2006 Subject: [Histonet] NYSHS Website Message-ID: Great Website Luis! WOOHOO !!!!!!!!!! GO NEW YORK !!! >>> Luis Chiriboga 01/27/06 01:17PM >>> Hi Everyone The New York State Histotechnology Society has just opened it's new website. This year, NYSHS is hosting the Region 1 Symposium and some preliminary information is available on the site. To visit the site just click on the following link: www.nyhisto.org Have a great weekend.... Luis ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Jan 27 19:26:57 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Jan 27 19:27:18 2006 Subject: [Histonet] Histonet's 10th anniversary! Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01305663@sjhaexc02.sjha.org> My thanks to you and Herb as well. Also to Marvin for the archives. And to all the members who are such a wealth of knowledge, I appreciate the community. Happy weekend to all! J:>) Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of LINDA MARGRAF Sent: Friday, January 27, 2006 5:50 PM To: Histonet@lists.utsouthwestern.edu Cc: hagler.herb@pathology.swmed.edu Subject: [Histonet] Histonet's 10th anniversary! Dear Histonet members: I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! There are currently 2342 members on the list from more than 30 different countries. I have no guess how many messages have gone through the server during this time. Must be millions. I'd like to personally thank all the members who actively participate in the list, making it such a worthwhile endeavor. Thanks so much. Thanks too to Herb for his behind the scenes computer expertise. Sincerely, Linda M Histonet administrator _______________________________________________ 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 katri <@t> cogeco.ca Fri Jan 27 21:32:14 2006 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Jan 27 21:32:21 2006 Subject: [Histonet] in vitro skin Fb staining w/vimentin References: Message-ID: <005401c623bb$6e8c41e0$6a9a9618@Katri> Hi Birthe, Vimentin is not specific marker for fibroblasts, although normal epithelial cells do not usually have that intermediate filament present. Many other cells do stain positively for vimentin, for instance melanocytes, macrophages and all lymphoid cells. Those are all present in the skin. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Birthe Schnegelsberg" To: Sent: Sunday, February 26, 2006 11:29 AM Subject: [Histonet] in vitro skin Fb staining w/vimentin > Hello. > > I am staining cells in in vitro skin with the fibroblast specific marker > vimentin to differentiate between fibroblasts and keratinocytes. > > I am able to detect fluorescent signals in the tissue. The staining > pattern > and the cell localization makes me wonder though, if the staining is in > fact > specific fibroblast staining or if the antibody (immunon) picks up dead > cell > material. > > Does anybody have experienece using vimentin as a marker for fibroblast > cells? > The staining pattern of the positive control looks different than that for > the singel cells in the in vitro skin?! > > Thanks, Birthe > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From katri <@t> cogeco.ca Fri Jan 27 21:35:50 2006 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Jan 27 21:35:56 2006 Subject: [Histonet] Histonet's 10th anniversary! References: Message-ID: <005b01c623bb$ef6a3790$6a9a9618@Katri> Thank you Linda and Herb for keeping it up and running. It is absolutely a great forum! Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "LINDA MARGRAF" To: Cc: Sent: Friday, January 27, 2006 5:50 PM Subject: [Histonet] Histonet's 10th anniversary! Dear Histonet members: I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! There are currently 2342 members on the list from more than 30 different countries. I have no guess how many messages have gone through the server during this time. Must be millions. I'd like to personally thank all the members who actively participate in the list, making it such a worthwhile endeavor. Thanks so much. Thanks too to Herb for his behind the scenes computer expertise. Sincerely, Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Sat Jan 28 00:32:32 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Sat Jan 28 00:32:43 2006 Subject: [Histonet] Histonet's 10th anniversary! References: Message-ID: <43DB1000.DB08408A@uwo.ca> Is it a coincidence that HistoNet's 10th birthday is also the 250th birthday of Wolfgang A. Mozart? Many happy returns! John Kiernan London, Canada ________________________________ LINDA MARGRAF wrote: > > Dear Histonet members: > I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! From jnocito <@t> satx.rr.com Sat Jan 28 09:09:45 2006 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Sat Jan 28 09:09:02 2006 Subject: [Histonet] Histonet's 10th anniversary! References: Message-ID: <007601c6241c$dfbbf840$e8bd0b43@yourxhtr8hvc4p> Dr. Margraf, it is us who thank you. You and your support staff have given us an information forum that has not only brought millions of pieces of advice, instructions and information, but has brought a world-wide community closer than any other source could have. Thank you!! Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "LINDA MARGRAF" To: Cc: Sent: Friday, January 27, 2006 4:50 PM Subject: [Histonet] Histonet's 10th anniversary! Dear Histonet members: I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! There are currently 2342 members on the list from more than 30 different countries. I have no guess how many messages have gone through the server during this time. Must be millions. I'd like to personally thank all the members who actively participate in the list, making it such a worthwhile endeavor. Thanks so much. Thanks too to Herb for his behind the scenes computer expertise. Sincerely, Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.1.375 / Virus Database: 267.14.23/243 - Release Date: 1/27/2006 From rjbuesa <@t> yahoo.com Sat Jan 28 09:49:24 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Jan 28 09:49:32 2006 Subject: [Histonet] Stains for Hemaglobin In-Reply-To: <21F1955FA284134E981948D6D3FFD04202ED3736@harpo.fhcrc.org> Message-ID: <20060128154924.18285.qmail@web61221.mail.yahoo.com> Julie: There are several methods for hemoglobin: 1- Ralph[Stain Technol., 16: 105 (1941)] for dry smears.Hemoglobin??dark brown. 2- Goulliart [C.reend.Soc.Biol.,135: 1260 (1941)] for frozen sections or dry smears. 3-Dunn [Arch.Pathol.,41: 676 (1946)] for frozen of paraffin sections of tissues fixed with NBF.Hemoglobin ?? bue; nuclei ??red; cytoplasm ?? pink. 4- O'Brien [Stain Technol. 36: 57 (1961) for for whole mount embryos, hematopietic cells and electrophoresis. Hope this will help you. Ren?? J. "Randolph-Habecker, Julie" wrote: I am looking for a specific stain for hemoglobin in formalin-fixed, paraffin-embedded canine tissue. I could either use IHC or a special stain. Any ideas? I found a reference for a Puchtler-Rosenthal-Sweat Method for hemoglobin but it says that it only works in tissue fixed in Zenker's. Just wondering if anyone has any experience in this before we try it. Thanks, Julie Julie Randolph-Habecker, Ph.D. Staff Scientist - Manager Experimental Histopathology Shared Resource Fred Hutchinson Cancer Research Center 1100 Fairview Ave, N. M5-A803 Seattle WA 98109-1024 206-667-6119 jhabecke@fhcrc.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- What are the most popular cars? Find out at Yahoo! Autos From mucram11 <@t> comcast.net Sat Jan 28 09:59:47 2006 From: mucram11 <@t> comcast.net (Pamela Marcum) Date: Sat Jan 28 09:59:51 2006 Subject: [Histonet] Histonet's 10th anniversary! In-Reply-To: <007601c6241c$dfbbf840$e8bd0b43@yourxhtr8hvc4p> Message-ID: <004101c62423$dd53a6a0$8d02a8c0@your4di1s53ime> I think Joe has said it all for us. We may have our days when we disagree or don't like what is said however, this is a service few of us would ever wish to be without. Like all groups if we all agreed on everything it would be boring and we would not be learning which is the biggest service of your work on this site. THANKS LINDA, HERB AND ALL WHO MAKE THIS WEBSITE POSSIBLE. Pam Marcum -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Saturday, January 28, 2006 10:10 AM To: LINDA MARGRAF; Histonet@lists.utsouthwestern.edu Cc: hagler.herb@pathology.swmed.edu Subject: Re: [Histonet] Histonet's 10th anniversary! Dr. Margraf, it is us who thank you. You and your support staff have given us an information forum that has not only brought millions of pieces of advice, instructions and information, but has brought a world-wide community closer than any other source could have. Thank you!! Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "LINDA MARGRAF" To: Cc: Sent: Friday, January 27, 2006 4:50 PM Subject: [Histonet] Histonet's 10th anniversary! Dear Histonet members: I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! There are currently 2342 members on the list from more than 30 different countries. I have no guess how many messages have gone through the server during this time. Must be millions. I'd like to personally thank all the members who actively participate in the list, making it such a worthwhile endeavor. Thanks so much. Thanks too to Herb for his behind the scenes computer expertise. Sincerely, Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.1.375 / Virus Database: 267.14.23/243 - Release Date: 1/27/2006 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMcCormick <@t> schosp.org Sat Jan 28 11:52:37 2006 From: JMcCormick <@t> schosp.org (McCormick, James) Date: Sat Jan 28 11:52:47 2006 Subject: [Histonet] NYSHS Website Message-ID: <913FAC2B773C19488E26AE6572180FA50315D283@exch01.schosp.org> DITTO,DITTO,DITTO......GREAT WEB SITE ! Wonderful service to the profession ! J.B.McCormick, M.D. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Vinnie Della Speranza Sent: Friday, January 27, 2006 5:54 PM To: Histonet@lists.utsouthwestern.edu; Luis.Chiriboga@med.nyu.edu Subject: Re: [Histonet] NYSHS Website Great Website Luis! WOOHOO !!!!!!!!!! GO NEW YORK !!! >>> Luis Chiriboga 01/27/06 01:17PM >>> Hi Everyone The New York State Histotechnology Society has just opened it's new website. This year, NYSHS is hosting the Region 1 Symposium and some preliminary information is available on the site. To visit the site just click on the following link: www.nyhisto.org Have a great weekend.... Luis ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 _______________________________________________ Histonet mailing list Histonet@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 Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. From pruegg <@t> ihctech.net Sat Jan 28 13:46:27 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Sat Jan 28 13:46:33 2006 Subject: [Histonet] Histonet's 10th anniversary! In-Reply-To: Message-ID: <200601281946.k0SJkLjH007918@chip.viawest.net> Linda and Herb, You are the one's to be thanked. I thank you both very much. I don't know how we ever functioned without this resource. Actually, I do remember and it was painful. I can remember feeling like I was the only person in the world trying to do such Histopathology as calcified bone in plastic, or IHC on bone and cartilage. Now we know that there our other's out there trying to solve the same problems we are, and in many cases the problems are already solved so we can get on with properly testing the sample at hand rather than developing the technique to do so. Best regards and here is to another 10 great years of HISTONET! Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of LINDA MARGRAF Sent: Friday, January 27, 2006 3:50 PM To: Histonet@lists.utsouthwestern.edu Cc: hagler.herb@pathology.swmed.edu Subject: [Histonet] Histonet's 10th anniversary! Dear Histonet members: I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! There are currently 2342 members on the list from more than 30 different countries. I have no guess how many messages have gone through the server during this time. Must be millions. I'd like to personally thank all the members who actively participate in the list, making it such a worthwhile endeavor. Thanks so much. Thanks too to Herb for his behind the scenes computer expertise. Sincerely, Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tahseen <@t> brain.net.pk Sat Jan 28 15:30:31 2006 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Sat Jan 28 23:33:01 2006 Subject: [Histonet] Histonet's 10th anniversary! References: Message-ID: <004501c62452$128d34c0$972bfea9@m7c0y4> Congratulations Histonet and to all of you who make this a great resource. Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Pakistan. ----- Original Message ----- From: LINDA MARGRAF To: Cc: Sent: Saturday, January 28, 2006 11:50 AM Subject: [Histonet] Histonet's 10th anniversary! Dear Histonet members: I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! There are currently 2342 members on the list from more than 30 different countries. I have no guess how many messages have gone through the server during this time. Must be millions. I'd like to personally thank all the members who actively participate in the list, making it such a worthwhile endeavor. Thanks so much. Thanks too to Herb for his behind the scenes computer expertise. Sincerely, Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From barry_m <@t> ozemail.com.au Sun Jan 29 06:16:43 2006 From: barry_m <@t> ozemail.com.au (Barry Madigan) Date: Sun Jan 29 06:16:58 2006 Subject: [Histonet] Histonets 10th Anniversary Message-ID: <000001c624cd$de4383a0$0201010a@WORKSTATION1> Congratulations to all who make the Histonet a wealth of information for the Histology community. It is certainly much appreciated. Well done and thank you. Barry Madigan Immunohistochemistry QHPS- RBH campus Australia From vanann702 <@t> skmc.gov.ae Sun Jan 29 07:03:51 2006 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Sun Jan 29 07:03:27 2006 Subject: [Histonet] well done Message-ID: <7E070A7B959A9F42BE732545E5CF6210948D0B@SKMCEMAIL.skmc.gov.ae> This source of info (and very often some serious entertainment!) is invaluable. I have been 'tuning in' since the very beginning and can't imagine running a lab with out being able to first 'check on the histonet' if I need a solution to a problem. Sometimes I just read and don't even have to write in myself - it is all there. We have so many problems in common - and all the handy hints and special 'tricks' are amazing. Keep it up - all of you - and thanks again for a wonderful site. Annieinarabia From Rcartun <@t> harthosp.org Sun Jan 29 07:57:40 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Sun Jan 29 07:58:18 2006 Subject: [Histonet] Histonet's 10th anniversary! Message-ID: Thank "YOU" Linda, Herb, and everyone else associated with Histonet. 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 >>> "LINDA MARGRAF" 01/27/06 05:50PM >>> Dear Histonet members: I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! There are currently 2342 members on the list from more than 30 different countries. I have no guess how many messages have gone through the server during this time. Must be millions. I'd like to personally thank all the members who actively participate in the list, making it such a worthwhile endeavor. Thanks so much. Thanks too to Herb for his behind the scenes computer expertise. Sincerely, Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Sun Jan 29 08:37:41 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Sun Jan 29 08:38:19 2006 Subject: [Histonet] Integrated Diagnostics, Inc. Message-ID: I recently uncovered some old literature from a company called "Integrated Diagnostics, Inc." located in Baltimore, MD. They manufactured products for infectious disease detection. Are they still operating? If not, does anyone know what happen to them? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From DonnaHansen <@t> chiwest.com Sun Jan 29 10:09:58 2006 From: DonnaHansen <@t> chiwest.com (Hansen, Donna (Ontario)) Date: Sun Jan 29 10:10:16 2006 Subject: [Histonet] Full time position for histologist. Message-ID: <91FAC8D14C98974F839CA842BBC67D42136364@nwont2dc01.ontario-or.catholichealth.net> Hello eveeryone, I wanted to let you know that Holy Rosary Medical Center located in Ontario Oregon has an immediate opening for a full time histologist. This is Monday through Friday, no weekends or holidays. We have competitive wages and benefits. Ontario is located about 50 miles north of Boise Idaho. We are on the Oregon/Idaho border. You can email me personally for more information. Thanks in advance! dohansen@chiwest.com 541-881-7289 Donna Hansen,HT(ASCP) Department Manager Holy Rosary Medical Center Ontario, Oregon 97914 From Rrlopez2 <@t> aol.com Sun Jan 29 12:18:18 2006 From: Rrlopez2 <@t> aol.com (Rrlopez2@aol.com) Date: Sun Jan 29 12:18:30 2006 Subject: [Histonet] Re: Histonet Digest, Vol 26, Issue 36 Message-ID: <26b.4d2a05d.310e60ea@aol.com> I just wanted to know if anyone out there is doing immuno double staining on a routine basis. If yes what detection kits and vendors are you using,any information would be helpful. Thank you in advance From jkiernan <@t> uwo.ca Sun Jan 29 13:10:36 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Sun Jan 29 13:10:46 2006 Subject: [Histonet] Re: cresyl violet staining vs. Haematoxylin staining {LONG} References: <43DA71B2.6080307@uci.edu> Message-ID: <43DD132C.9AA331B6@uwo.ca> This is an answer to two Histonet questions asked on 27th Jan 2006. It's addressed primarily to Chengkang Zhang, who asked about staining neurons with haemalum ("haematoxylin") or cresyl violet. The other question, from MOHANA was about the mechanism of Nissl staining by cresyl violet. The answers for MOHANA are mostly in the next paragraph but one. Assuming that you (Zhang) are using a haemalum (alum-haematein): this stains the nuclei of all cells, including neurons and neuroglia. There are other haematoxylin-based stains that are very selective for nuclei, ones that stain myelin, and all sorts of others. Haemalum is the H in H&E. Cresyl violet, on the other hand, is a cationic ("basic") dye that attaches to anionic macromolecules. In the central nervous system, nuclear DNA and ribosomal RNA (rRNA) are the only such materials. DNA is in the nuclei of all cells: neurons and neuroglia. All cell nuclei are stained by cresyl violet, just as they are (though for a different reason) by haemalum. RNA is present in the cytoplasm of all cells, but neurons contain much more rRNA than glial cells. In the cytoplasm around a neuron's nucleus (the perikaryon), the rRNA is often organized into stripes or granules, characteristic of the type of neuron, called Nissl bodies. (Messenger RNA and transfer RNA probably doesn't contribute to the picture; that's another story.) Nissl didn't use cresyl violet, by the way; he used methylene blue. (In the December 2005 issue of Microscopy Today, on p.50, there's a historical note about Franz Nissl.) Large and medium-sized neurons and the main glial cell types can be identified from the appearances of their nuclei (stained with either haemalum or a cationic dye). Some small neurons have densely staining nuclei and cannot be certainly distinguished from oligodendrocytes in stained sections. Among the granule cells of the cerebellum or olfactory bulb, or in layer 4 of a sensory area of the cerebral cortex, individual cells cannot be categorized on the strength of nuclear appearance. In such regions the cytoplasmic Nissl bodies are not very helpful. A small interneuron has only a little stainable cytoplasmic rRNA. Undisputed oligodendrocytes (in white matter) also have subtle rings of rRNA around their nuclei. This glial cytoplasmic rRNA is distinct from the DNA subjacent to the nuclear membrane, and it stains red (RNA) not blue-green (DNA) with the ethyl green-pyronine method. I've seen this myself, long ago, but cannot conjure up the even older original refs. If your library has "Methods in Brain Research" edited by P.B.Bradley (Wiley, 1975), you may find the ref in Chapter 1. John Kiernan Anatomy, UWO London, Canada ________________________________________ Chengkang ZHANG wrote: > > I am currently using haematoxylin to stain nuclei of neurons, but > someone told me that cresyl violet is better than haematoxylin. Would > anyone tell me what the difference between those two staining and which > one you prefer for nuclei staining? > > Thanks > > Chengkang > -- > =================================== > Chengkang Zhang Ph.D. > Department of Pharmacology > University of California, Irvine > Irvine, CA 92697-4625 > Email: chengkaz@uci.edu > TEL: (949)-824-1902 (lab) > FAX: (949)-824-4855 > =================================== MOHANA wrote: > > i was wondering if any one can explain me that why does the cresyl violet stains only neurons and not glia???? > > mohana > > Mohana _________________________________________ From asmith <@t> mail.barry.edu Sun Jan 29 16:12:30 2006 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Sun Jan 29 16:12:39 2006 Subject: [Histonet] 10th Anniversary Message-ID: <5D2189E74151CC42BEC02906BA8996322B916A@exchsrv01.barrynet.barry.edu> Congratulations and many, many thanks to our hosts at U. Texas Southwestern! Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From AnthonyH <@t> chw.edu.au Sun Jan 29 16:18:40 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Sun Jan 29 16:22:08 2006 Subject: [Histonet] Histonet's 10th anniversary! Message-ID: Brilliant!! Thanks wholeheartedly A very much appreciated job. 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----- ********************************************************************** 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: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of LINDA MARGRAF Sent: Saturday, 28 January 2006 9:50 AM To: Histonet@lists.utsouthwestern.edu Cc: hagler.herb@pathology.swmed.edu Subject: [Histonet] Histonet's 10th anniversary! Dear Histonet members: I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! There are currently 2342 members on the list from more than 30 different countries. I have no guess how many messages have gone through the server during this time. Must be millions. I'd like to personally thank all the members who actively participate in the list, making it such a worthwhile endeavor. Thanks so much. Thanks too to Herb for his behind the scenes computer expertise. Sincerely, Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Sun Jan 29 16:22:59 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Sun Jan 29 16:25:15 2006 Subject: [Histonet] Histonet's 10th anniversary! Message-ID: Mozart? Is he the bloke who developed Mozart's Pentachrome Stain? Err No that was Movat's Pentachrome!! Sorry 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----- ********************************************************************** 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: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Saturday, 28 January 2006 5:33 PM To: LINDA MARGRAF Cc: Histonet@lists.utsouthwestern.edu; hagler.herb@pathology.swmed.edu Subject: Re: [Histonet] Histonet's 10th anniversary! Is it a coincidence that HistoNet's 10th birthday is also the 250th birthday of Wolfgang A. Mozart? Many happy returns! John Kiernan London, Canada ________________________________ LINDA MARGRAF wrote: > > Dear Histonet members: > I wanted to let everyone know that this week is the 10th anniversary > of Histonet. Its hard to believe its been going strong now for 10 > years! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From llewllew <@t> shaw.ca Sun Jan 29 17:13:09 2006 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Sun Jan 29 17:14:06 2006 Subject: [Histonet] Histonet 10th anniversary Message-ID: <000501c62529$91bac0a0$690a4246@yourlk4rlmsu> The Histonet has obviously become one of the most successful international mediums for histotechs, and you should be congratulated. To those in the NSH, why not consider a special Golden Forceps award to those resonsible for it? Bryan Llewellyn From jkiernan <@t> uwo.ca Sun Jan 29 23:33:31 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Sun Jan 29 23:33:42 2006 Subject: [Histonet] Histonet's 10th anniversary! References: Message-ID: <43DDA52B.A8248EEC@uwo.ca> That's the one Tony. You weigh it out on a chromatic scale, and don't write too many notes ... John Kiernan London, Canada. -------------------------------------- Tony Henwood wrote: > > Mozart? > > Is he the bloke who developed Mozart's Pentachrome Stain? > > Err > No that was Movat's Pentachrome!! > > Sorry > > Regards > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Locked Bag 4001, Westmead, 2145, AUSTRALIA. > > -----Original Message----- > > Is it a coincidence that HistoNet's 10th birthday > is also the 250th birthday of Wolfgang A. Mozart? > > Many happy returns! > > John Kiernan > London, Canada > ________________________________ > LINDA MARGRAF wrote: > > > > Dear Histonet members: > > I wanted to let everyone know that this week is the 10th anniversary > > of Histonet. Its hard to believe its been going strong now for 10 > > years! From jkiernan <@t> uwo.ca Sun Jan 29 23:36:18 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Sun Jan 29 23:36:27 2006 Subject: [Histonet] Histonet's 10th anniversary! References: Message-ID: <43DDA5D2.6C601296@uwo.ca> That's the one Tony. You weigh it out on a chromatic scale, and don't write too many notes ... John Kiernan London, Canada. -------------------------------------- Tony Henwood wrote: > > Mozart? > > Is he the bloke who developed Mozart's Pentachrome Stain? > > Err > No that was Movat's Pentachrome!! > > Sorry > > Regards > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Locked Bag 4001, Westmead, 2145, AUSTRALIA. > > -----Original Message----- > > Is it a coincidence that HistoNet's 10th birthday > is also the 250th birthday of Wolfgang A. Mozart? > > Many happy returns! > > John Kiernan > London, Canada > ________________________________ > LINDA MARGRAF wrote: > > > > Dear Histonet members: > > I wanted to let everyone know that this week is the 10th anniversary > > of Histonet. Its hard to believe its been going strong now for 10 > > years! From JMacDonald <@t> mtsac.edu Sun Jan 29 23:47:15 2006 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Sun Jan 29 23:50:45 2006 Subject: [Histonet] CSH Annual Symposium Message-ID: The California Society for Histotechnology will be having their a nnual symposium at the Costa Mesa Hilton (Southern California) from May 18- be CMP. If you would like more information please contact: &nb References 1. 3D"mailto:jmacdonald@mtsac.edu" From AnthonyH <@t> chw.edu.au Mon Jan 30 00:30:57 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Mon Jan 30 00:31:08 2006 Subject: [Histonet] Histonet's 10th anniversary! Message-ID: Hee, Hee, Hee ! :) 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 John Kiernan Sent: Monday, 30 January 2006 4:34 PM To: Histonet Subject: Re: [Histonet] Histonet's 10th anniversary! That's the one Tony. You weigh it out on a chromatic scale, and don't write too many notes ... John Kiernan London, Canada. -------------------------------------- Tony Henwood wrote: > > Mozart? > > Is he the bloke who developed Mozart's Pentachrome Stain? > > Err > No that was Movat's Pentachrome!! > > Sorry > > Regards > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Locked Bag > 4001, Westmead, 2145, AUSTRALIA. > > -----Original Message----- > > Is it a coincidence that HistoNet's 10th birthday > is also the 250th birthday of Wolfgang A. Mozart? > > Many happy returns! > > John Kiernan > London, Canada > ________________________________ > LINDA MARGRAF wrote: > > > > Dear Histonet members: > > I wanted to let everyone know that this week is the 10th anniversary > > of Histonet. Its hard to believe its been going strong now for 10 > > years! _______________________________________________ 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 mtyler <@t> uctgsh1.uct.ac.za Mon Jan 30 02:03:03 2006 From: mtyler <@t> uctgsh1.uct.ac.za (Tyler) Date: Mon Jan 30 02:03:16 2006 Subject: [Histonet] CD antibodies Message-ID: <43DDC837.AB9FA178@uctgsh1.uct.ac.za> Morning Please could you advise which companies supply CD antibodies against rabbit tissue. Thank You Marilyn From kemlo.rogerson <@t> waht.swest.nhs.uk Mon Jan 30 02:47:24 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Jan 30 02:47:33 2006 Subject: [Histonet] Gyn cytologies Message-ID: Liquid based gynae cytology or conventional gynae cytology? If conventional SurePath or Cytyc? I would certainly recommend LBC over conventional but the choice of technology may be controversial. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Histology SLU [mailto:sluhisto@yahoo.com] Sent: Friday, January 27, 2006 10:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Gyn cytologies Hello All: Our lab may be getting into the gyn cytology business staining pap smears. It has been many, many, many years since I have done this. I'm sure that there are many advances. If any of you are doing gyn cytologies in your labs, would you mind sharing your expertise with me? Are you manual or is there automation for staining pap smears? What companies do you get your staining reagents from, etc? Thanks so much. You are all always a wealth of info. Have a super weekend!! Susan --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jtrynak <@t> hotmail.com Mon Jan 30 06:51:15 2006 From: jtrynak <@t> hotmail.com (John Rynak) Date: Mon Jan 30 06:51:29 2006 Subject: [Histonet] Wanted: Sales Representative - Histology Equipment - Eastern Canada - Toronto Message-ID: SciGenium's Client is experiencing tremendous growth within their North American operations. This growth is due to the recent launch of their FDA approved automated histology instrumentation system, and the growing antibody and reagent consumable product line for the histology market. Due to the new launch of these products they are currently looking for Senior Account Representatives, within the following region Eastern Canada. The ideal candidate will have had 3+ years of experience selling capital equipment and associated consumables into a clinical diagnostics laboratory. This person will have had prior experience selling to end user or laboratory manager ideally within the histopathology sector, and a BS/MS in a life science discipline. This position requires the ability to travel 75% of the time in a multistate region. Interested parties should forward a resume to resume@scigenium.com Send to : SciGenium PO 380916 Cambridge, MA 02238 From bruski33 <@t> aol.com Mon Jan 30 10:19:01 2006 From: bruski33 <@t> aol.com (bruski33@aol.com) Date: Mon Jan 30 10:19:16 2006 Subject: [Histonet] research blocks Message-ID: <8C7F3D003F58E23-1378-7A2A@FWM-R06.sysops.aol.com> Histo tech's with 30 yrs experience looking for research blocks to cut. Will cut paraffin blocks for unstained or H&E's rate is negotiable. We are in MA but will do blocks from any where in USA. contact Bruski33@aol.com From dmarsha3 <@t> utmem.edu Mon Jan 30 10:26:49 2006 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Mon Jan 30 10:27:08 2006 Subject: [Histonet] Histonet's 10th anniversary! References: Message-ID: <006801c625b9$f8b692d0$f623c084@DanaM> Thanks to you and to everyone who has been so helpful to me (clearly a novice with my novice questions) through this service. Dana ----- Original Message ----- From: "LINDA MARGRAF" To: Cc: Sent: Friday, January 27, 2006 4:50 PM Subject: [Histonet] Histonet's 10th anniversary! Dear Histonet members: I wanted to let everyone know that this week is the 10th anniversary of Histonet. Its hard to believe its been going strong now for 10 years! There are currently 2342 members on the list from more than 30 different countries. I have no guess how many messages have gone through the server during this time. Must be millions. I'd like to personally thank all the members who actively participate in the list, making it such a worthwhile endeavor. Thanks so much. Thanks too to Herb for his behind the scenes computer expertise. Sincerely, Linda M Histonet administrator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Mon Jan 30 10:39:21 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Jan 30 10:39:53 2006 Subject: [Histonet] Stains for Hemaglobin Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171765A@lsexch.lsmaster.lifespan.org> Years ago I had to do a hemoglobin stain, to demonstrate that round inclusions in some large cells were in fact phagocytized erythrocytes (thereby proving that the cells in question were parasitic amebae). I used one of two techniques I found in an old book (Histopathologic Technic and Practical Histochemistry by Lillie and Fullmer, 1976), and whichever technique I used was quite simple and worked very well. I'm not sure which one I used (I think it was Dunn-Thompson) so I'll give you both ... DUNN-THOMPSON METHOD 1. Stain 15 minutes in Mallory's aqueous alum hematoxylin (or other unacidified alum hematoxylin) 2. Wash in tap water 3. Mordant 1 minute in 4% iron alum 4. Rinse in tap water 5. Stain 15 minutes in picrofuchsin solution (13 ml 1% acid fushsin in 87 ml saturated aqueous picric acid) 6. Differentiate 3 minutes in 95% ethanol 7. Dehydrate, clear, mount Results: Hemoglobin emerald green; cytoplasm brown to yellow; collagen red; nuclei purple to gray-black OKAJIMA METHOD 1. Mordant 1 minute in 10% phosphomolybdic acid 2. Wash in distilled water 3. Stain 1 hour in ... 9 ml 10% phosphomolybdic acid in 30 ml saturated aqueous alizarin red 4. Wash in distilled water 5. Counterstain in an unacidified alum hematoxylin , 3 to 5 minutes 6. Wash in water, dehydrate, clear, mount Results: Hemoglobin orange-red; background light brown From jengirl1014 <@t> yahoo.com Mon Jan 30 12:10:41 2006 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Mon Jan 30 12:10:53 2006 Subject: [Histonet] Re: Histonet's 10th anniversary Message-ID: <20060130181041.62994.qmail@web60617.mail.yahoo.com> Thanks to the creators of the website for putting this together. It's helped me several times with beginners stuff. Not once has anyone made me feel like I was inferior. You all have been very generous over the past 2 years and I rellay appreciate it! Happy 10th and may there be gazillions more! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 410-955-9688 e-mail: jengirl1014@yahoo.com --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From Luis.Chiriboga <@t> med.nyu.edu Mon Jan 30 12:27:35 2006 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Mon Jan 30 12:24:25 2006 Subject: [Histonet] NYSHS Website In-Reply-To: <913FAC2B773C19488E26AE6572180FA50315D283@exch01.schosp.org> Message-ID: Hi Guys I wish I could take the credit......The site was designed by someone else!! I was just the messenger :-) Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of McCormick, James Sent: Saturday, January 28, 2006 12:53 PM To: Vinnie Della Speranza; Histonet@lists.utsouthwestern.edu; Luis.Chiriboga@med.nyu.edu Subject: RE: [Histonet] NYSHS Website DITTO,DITTO,DITTO......GREAT WEB SITE ! Wonderful service to the profession ! J.B.McCormick, M.D. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Vinnie Della Speranza Sent: Friday, January 27, 2006 5:54 PM To: Histonet@lists.utsouthwestern.edu; Luis.Chiriboga@med.nyu.edu Subject: Re: [Histonet] NYSHS Website Great Website Luis! WOOHOO !!!!!!!!!! GO NEW YORK !!! >>> Luis Chiriboga 01/27/06 01:17PM >>> Hi Everyone The New York State Histotechnology Society has just opened it's new website. This year, NYSHS is hosting the Region 1 Symposium and some preliminary information is available on the site. To visit the site just click on the following link: www.nyhisto.org Have a great weekend.... Luis ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 _______________________________________________ Histonet mailing list Histonet@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 Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From maria <@t> ski.org Mon Jan 30 12:24:43 2006 From: maria <@t> ski.org (Maria Mejia) Date: Mon Jan 30 12:24:55 2006 Subject: [Histonet] congratulations to all Message-ID: <43DE59EB.3060405@ski.org> Dear Histonet, To all the histonet people from every corner of the world, my deepest appreciation and gratitude for making this listserver a priceless treasure of information. What a great feeling it is to know that we are not alone in our work and equally important in those times when we need help and guidance. Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 From DEllenburg2 <@t> stfrancishealth.org Mon Jan 30 12:31:52 2006 From: DEllenburg2 <@t> stfrancishealth.org (DEllenburg2@stfrancishealth.org) Date: Mon Jan 30 12:28:32 2006 Subject: [Histonet] Extended Region III Hotel Deadline Message-ID: <0A502E8156DAA4468CB8979B27177555016CC580@bssmsx5501.intranet.stfrancishealth.com> There has been a "GLITCH" with the extended date to make your hotel reservations for the Region III Meeting. The newest deadline for making reservations for the Region III Meeting is Friday February 3rd. If you call the toll free number 888-485-0136 and have problems concerning the newest deadline of February 3rd you will need to call 843-284-7026 and ask for Tracy in sales. If additional help is needed you can call Linda Jenkins at 864- 238-8230 or 864-656-5553. I apologize for any inconvenience that this might have caused anybody. Hope to see you there. Deborah Ellenburg, HT (ASCP) Bon Secours St. Francis Health System One St. Francis Drive Greenville, SC 29601 864-255-1582 The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at (864) 255-1000, and permanently delete the original e-mail, attachment(s), and any copies. From dsantana <@t> pmaonline.com Mon Jan 30 12:36:29 2006 From: dsantana <@t> pmaonline.com (Santana, Diane) Date: Mon Jan 30 12:42:42 2006 Subject: [Histonet] (no subject) Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB07113821@MAILPMA> I have a question for any other small lab. Up to now I only do small GI biopsies, we will be getting some derm and in the future some larger specimens. How do I set up my tissue processor for large specimen without ruin my small biopsies? Also has anyone else had problems with using different alcohols from different vendors. Example if I use an Alcohol with Isopropyl, Ethel, methyl alcohol, to a alcohol with only isopropyl or some other combo do you need to readjust your times in the last 100% alcohols. Thanks Diane Santana PMA Haverhill, Mass. From PMonfils <@t> Lifespan.org Mon Jan 30 13:10:27 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Jan 30 13:10:46 2006 Subject: [Histonet] (dehydration question) Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171765B@lsexch.lsmaster.lifespan.org> I would suggest processing your larger specimens on an overnight schedule, and your small biopsies on a daytime schedule (the entire process taking anywhere from 4 to 6 hours). There may be minor differences in how rapidly various alcohols remove water from tissue, but probably not great enough to warrant changing your dehydration times. However, you should be aware that the different alcohols do have some different effects on tissue. For example, isopropanol causes less tissue shrinkage and less tissue hardening than ethanol does. This characteristic doesn't make much practical difference for most tissues, but it could be either an advantage or a disadvantage for certain tissues. Methanol dehydrates faster than the other two, but I would avoid products containing it because of its greater toxicity. It can be absorbed through the skin, and its vapors are also toxic. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Santana, Diane > Sent: Monday, January 30, 2006 10:36 AM > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] (no subject) > > I have a question for any other small lab. Up to now I only do small GI > biopsies, we will be getting some derm and in the future some larger > specimens. How do I set up my tissue processor for large specimen without > ruin my small biopsies? Also has anyone else had problems with using > different alcohols from different vendors. Example if I use an Alcohol > with > Isopropyl, Ethel, methyl alcohol, to a alcohol with only isopropyl or some > other combo do you need to readjust your times in the last 100% alcohols. > Thanks > Diane Santana > PMA > Haverhill, Mass. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From kaleid11 <@t> yahoo.com Mon Jan 30 15:52:07 2006 From: kaleid11 <@t> yahoo.com (Adam Perry) Date: Mon Jan 30 15:52:16 2006 Subject: [Histonet] brown nickel-DAB staining... Message-ID: <20060130215207.73069.qmail@web30413.mail.mud.yahoo.com> Has anyone ever performed immunocytochemistry using nickel-DAB as the chromogen in the peroxidase step and gotten brown specific staining (similar to DAB) and the usual purple-black nonspecific background staining in the tissue (characteristic of excessive nickel-DAB reaction product)? Here's a little more background: I have site-specifically injected an adeno-associated viral vector that expresses human estrogen receptor into rodent brains that normally express rodent estrogen receptor. When I performed one round of ICC using an antibody that recognizes both human and rodent estrogen receptor, I obtained black (Ni-DAB) stained cell nuclei in regions throughout the brain that normally express estrogen receptor and brown stained nuclei in the area that I injected the viral vector (and presumably contains cells expressing the human estrogen receptor). I then performed ICC using a monoclonal antibody that recognizes human estrogen receptor (rodent specificity hasn't been determined for this antibody yet)...and I only get brown staining in the area I injected with the viral vector- even though I used Ni-DAB as the chromagen (so should have been black)... I'm thinking maybe the viral expression of the human receptor is so high that there are huge amounts of my protein..and maybe that is somehow affecting the Ni-DAB reaction...but I feel like that's a stretch...has anyone ever gotten similar results (i.e BROWN signal from Ni-DAB with black background staining??) Any insights would be greatly appreciated.... Thanks, Adam --------------------------------- What are the most popular cars? Find out at Yahoo! Autos From rjbuesa <@t> yahoo.com Mon Jan 30 16:37:08 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Jan 30 16:37:16 2006 Subject: [Histonet] brown nickel-DAB staining... In-Reply-To: <20060130215207.73069.qmail@web30413.mail.mud.yahoo.com> Message-ID: <20060130223708.47472.qmail@web61217.mail.yahoo.com> Adam: I am not sure if what I am going to tell you will help or not but, there it goes! I used 3 drops (about 120 ?L) of a 2% aq. sol. of nickel sulfate [NiSO4.6H2O] + 4 drops (about 160 ?L) of 3% H2O2 + 10 mL of PBS + 6 mg of DAB as a second chromogen (permanent) when I did a 2 antigen detection on the same section. I let react x 6 minutes. The first chromogen was the regular DAB and the developed colour with the Ni-DAB was always purplishblue, never brown or black. Both colours where present in the 2 different antigen sites, and never as a background. To me it seems that you have some dilution to fiddle with. Hope this will help after all! Ren? J. Adam Perry wrote: Has anyone ever performed immunocytochemistry using nickel-DAB as the chromogen in the peroxidase step and gotten brown specific staining (similar to DAB) and the usual purple-black nonspecific background staining in the tissue (characteristic of excessive nickel-DAB reaction product)? Here's a little more background: I have site-specifically injected an adeno-associated viral vector that expresses human estrogen receptor into rodent brains that normally express rodent estrogen receptor. When I performed one round of ICC using an antibody that recognizes both human and rodent estrogen receptor, I obtained black (Ni-DAB) stained cell nuclei in regions throughout the brain that normally express estrogen receptor and brown stained nuclei in the area that I injected the viral vector (and presumably contains cells expressing the human estrogen receptor). I then performed ICC using a monoclonal antibody that recognizes human estrogen receptor (rodent specificity hasn't been determined for this antibody yet)...and I only get brown staining in the area I injected with the viral vector- even though I used Ni-DAB as the chromagen (so should have been black)... I'm thinking maybe the viral expression of the human receptor is so high that there are huge amounts of my protein..and maybe that is somehow affecting the Ni-DAB reaction...but I feel like that's a stretch...has anyone ever gotten similar results (i.e BROWN signal from Ni-DAB with black background staining??) Any insights would be greatly appreciated.... Thanks, Adam --------------------------------- What are the most popular cars? Find out at Yahoo! Autos _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with your Yahoo! Mail. From pruegg <@t> ihctech.net Mon Jan 30 17:05:48 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Jan 30 17:05:52 2006 Subject: [Histonet] Spring meeting in CO Message-ID: <200601302305.k0UN5ejH027596@chip.viawest.net> COLORADO SOCIETY FOR HISTOTECHNOLOGY The CSH will be hosting their 2006 Spring Symposium on April 28th and 29th of 2006 at the Cheyenne Mountain Resort. It is located at 3225 Broadmoor Valley Road, Colorado Springs Co 80906 1-800-428-8886. We have acquired quite the group of speakers for this year including: Mr. Skip Brown, Dr. Stephan Cina, Mr. Lance Erickson, Ms. Janet Maass, Mrs. Debra Flynn and Mr. Mike Reichenbach of Ventana Medical Systems, Ms. Charlie Dorner of Vision Biosytems, and Mrs. Konnie Zeitner. We will have workshops from 1:00 pm to 4:30 pm on Friday the 28th and from 8:30 am to 4:30 pm on Sat the 29th. A social will be held on Friday evening from 5pm-7pm. Linda Schriner will be in charge of registration this year and our vendor contact is Rick Garnhart. Contact info for Linda Schriner Contact info for Rick Garnhart 1285 Blackhawk RD Memorial Hospital Histology Eaton Co 80615 1400 E Boulder 970 350-6427 W Colorado Springs, CO 80909 970 454-3727 H Ph: (719) 365-5204 970 350-6477 F Fx: (719) 365-6373 linda.schriner@bannerhealth.com rick.garnhart@memhospcs.org Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From sharon.osborn <@t> dnax.org Mon Jan 30 15:56:51 2006 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Mon Jan 30 17:06:19 2006 Subject: [Histonet] histonet and W.A. Mozart! Message-ID: <29B25753F6B1D51196110002A589D444032C49D5@PALMSG30.us.schp.com> May sharing a birthday with Wolfgang Amadeus Mozart ensure a long lived histonet! John, you are a man after my heart! The radio station I listen to, KDFC-FM and www.KDFC.com streaming celebrated WAM's life with his music through his ages last week! What great listening it made. sharon osborn DNAX, SP BioPharma Palo Alto, CA > Is it a coincidence that HistoNet's 10th birthday > is also the 250th birthday of Wolfgang A. Mozart? > > Many happy returns! > > John Kiernan > London, Canada ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From smclaugh <@t> fhcrc.org Mon Jan 30 17:15:56 2006 From: smclaugh <@t> fhcrc.org (McLaughlin, Sharon R) Date: Mon Jan 30 17:16:30 2006 Subject: [Histonet] Giemsa staining on Bone Marrow smears Message-ID: <8BD501B158A381409FE54DB421F82FCA02E8B6A1@groucho.fhcrc.org> Hi, This is in response to a question about the Giemsa staining on bone marrow smears. First of all, the procedure for making the smears has to be ideal or it will not be the best staining. Make sure whomever is doing the smears is using the right technique. That can make all the difference in the world. I use the May-Gruwald Giemsa Stain from R.D. Lillie. Reagents: Stock Giemsa Solution: Giemsa powder .............................................1.0 gm Glycerin......................................................66.0 ml Absolute Methanol....................................... 66.0 ml *********Mix the giemsa powder and the glycerin and place in a 60 degree oven for 2 hours. After the incubation, add the 66 ml of absolute methanol. Mix well. Working Giemsa Solution: Stock Giemsa Solution.................................40 drops Distilled water.............................................40 ml *********Make fresh, do not reuse 1% Glacial Acetic Acid: Concentrated Glacial Acetic Acid..................1.0 ml Distilled water....................................... 99.0 ml Stock Jenner: Jenner stain, dry powder.............................1.0 gm Absolute methanol.................................400.0 ml Working Jenner: Stock Jenner solution...............................25.0 ml Distilled water.........................................25.0 ml Procedure: 1. Deparaffinize and hydrate to tap water. 2. Wash in running water for 10 minutes. 3. Rinse in distilled water, 2 changes. 4. Place in absolute methyl alcohol, 2 changes, 3 minutes each. 5. Place in working Jenner solution for 6 minutes. 6. Place in working Giemsa solution for 45 minutes. 7. IMPORTANT STEP: Handle each slide individually in this and subsequent steps. Rinse quickly in distilled water - 1 dip. 8. Differentiate in 1% glacial acetic acid just until the sections becomes pink!!!!!!!. Check microscopically for well differentiated nuclei. Red cells will become reddish pink. 9. Rinse well in several changes in distilled water. 10. Dehydrate quickly, clear and mount. Results: Mast cell granules.....................................Rust red Nuclei.................................................... Blue Cytoplasm...............................................Pink to rose Bacteria and spirochetes............................Rose to purple Rickettsiae............................................. Violet Chromatin of parasites.............................. Red RBC's................................................... Salmon red From marjoh3 <@t> telus.net Mon Jan 30 21:08:39 2006 From: marjoh3 <@t> telus.net (Marilyn Johnson) Date: Mon Jan 30 21:08:47 2006 Subject: [Histonet] Immunohistochemistry on Poultry Tissues Message-ID: <001801c62613$a2286bf0$6401a8c0@VALUED20606295> Hi Histonetters, If anyone performs IHC on poultry tissues please give me a list of your tests. I am particularily interested in testing for Adeno virus in turkeys. Thanks in advance to any replies to my request. Marilyn Johnson Alberta Agriculture Edmonton, Alberta, Canada, From sjchtascp <@t> yahoo.com Tue Jan 31 08:01:18 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Tue Jan 31 08:01:27 2006 Subject: [Histonet] FFPE control Message-ID: <20060131140118.49616.qmail@web90210.mail.scd.yahoo.com> Good morning all, I'd like to pick up a couple of control blks if anyone has some to spare. They are pancreas and gram +/-. I also have almost 2 cases of surgipath + slides, white and lavender for trade or for anyone that need them. The scientist I work for only wants me to us VWR slides. Just let me know. Steve --------------------------------- Do you Yahoo!? With a free 1 GB, there's more in store with Yahoo! Mail. From dgaupp <@t> tulane.edu Tue Jan 31 11:33:40 2006 From: dgaupp <@t> tulane.edu (dgaupp@tulane.edu) Date: Tue Jan 31 11:33:46 2006 Subject: [Histonet] HELP! vertical lines in paraffin Message-ID: <1138728820.43df9f74bfb87@webmail.tulane.edu> Histonetters: I am having problems sectioning mouse ovary tumor without vertical lines from the bottom of the block(paraffin only - no tissue touching the blade) all the way to top of block. I did not process/embed these tissues. So far I've done the following: 1. changed blades(went threw about 10 just in a few blocks) 2. cleaned microtome 3. changed angles(6-12 degree & everything in between) 4. sectioned hot, cold, freezing 5. soaked blocks in ice cold water; froze blocks 6. surfaced decal 5 - 40 minute intervals 7. re-embedded The paraffin seems to be causing the lines! I've never experienced problems like this before. Usually its the tissue itself that causes a nick in blade which therefore causing vertical lines to appear when sectiong the tissue. Help is needed! What next is recommended? Dina Gaupp, BS MT Tulane University Medical Center Center for Gene Therapy Histology Core Facility 1430 Tulane Ave New Orleans, La 70112 Lab phone 504.988.1194 email dgaupp@tulane.edu From victor <@t> pathology.washington.edu Tue Jan 31 11:34:49 2006 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Tue Jan 31 11:34:56 2006 Subject: [Histonet] Barcoding Cassettes Message-ID: <43DF9FB9.9090508@pathology.washington.edu> Any facilities out there that are routinely barcoding cassettes? What problems have you experienced with either the performance of the printer or the readability of the barcode after processing? 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 PMonfils <@t> Lifespan.org Tue Jan 31 12:15:17 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Tue Jan 31 12:15:29 2006 Subject: [Histonet] HELP! vertical lines in paraffin Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171765C@lsexch.lsmaster.lifespan.org> Dirt in the paraffin reservoir of the embedding unit can cause such problems, but it would really have to be quite dirty to cause the extent of problem you describe. Have you tried melting the blocks and re-embedding them in your own paraffin? When I first started in histology we didn''t have the quality of commecial products we have today. We kept a large flask and funnel in the oven at all times, with filter paper in the funnel. Newly opened paraffin was poured into the funnel and allowed to melt and filter before use, and it was amazing the amount of solid contaminants the filter paper would collect. If we used the paraffin without filtering it, we would get the same kind of problems you are describing. Today's commercial products are pre-filtered and much purer. But a poorly maintained embedding unit can intruduce contaminants into the paraffin. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > dgaupp@tulane.edu > Sent: Tuesday, January 31, 2006 9:33 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] HELP! vertical lines in paraffin > > Histonetters: > > > I am having problems sectioning mouse ovary tumor without vertical lines > from > the bottom of the block(paraffin only - no tissue touching the blade) all > the > way to top of block. I did not process/embed these tissues. So far I've > done > the following: > > 1. changed blades(went threw about 10 just in a few blocks) > 2. cleaned microtome > 3. changed angles(6-12 degree & everything in between) > 4. sectioned hot, cold, freezing > 5. soaked blocks in ice cold water; froze blocks > 6. surfaced decal 5 - 40 minute intervals > 7. re-embedded > The paraffin seems to be causing the lines! I've never experienced > problems > like this before. Usually its the tissue itself that causes a nick in > blade > which therefore causing vertical lines to appear when sectiong the tissue. > Help is needed! What next is recommended? > > > > Dina Gaupp, BS MT > Tulane University Medical Center > Center for Gene Therapy > Histology Core Facility > 1430 Tulane Ave > New Orleans, La 70112 > Lab phone 504.988.1194 > email dgaupp@tulane.edu > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gcallis <@t> montana.edu Tue Jan 31 12:35:20 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Jan 31 12:35:35 2006 Subject: [Histonet] HELP! vertical lines in paraffin In-Reply-To: <1138728820.43df9f74bfb87@webmail.tulane.edu> References: <1138728820.43df9f74bfb87@webmail.tulane.edu> Message-ID: <6.0.0.22.1.20060131110430.01b37840@gemini.msu.montana.edu> Dina, We experienced what you have described with outside sourced blocks and could never get rid of the ski tracks. We finally ended up melting the tissue out of block and reembedding in our own paraffin, this alleviated the problem somewhat but not 100%. It may be they do not stir the paraffin to redistribute the plastic polymers i.e additives before embedding and/or do not clean out their dispenser. Their paraffin may be really old too?? Some paraffins seem to suffer more from additives settling out. Usually during automated processing, agitation takes care of the settling out problem. Some labs are also careless about cleaning paraffin dispensers regularly. One observation made here some years ago with our own paraffin dispenser. Years ago, we never stirred the paraffin before embedding, and when we finally got round to cleaning the dispenser, there was a layer of gunky plastic appearing junk stuck to bottom of the dispenser. Well, we now stir before embedding and clean dispenser regularly. If it persists from this lab, then processing and embedding the tissue in your lab may be the best recourse and this is our policy now. They should be informed what is happening and what the problems could be and hopefully this will be corrected. Curious thought --- what kind of sectioning problems do they have? At 10:33 AM 1/31/2006, you wrote: >I am having problems sectioning mouse ovary tumor without vertical lines from >the bottom of the block(paraffin only - no tissue touching the blade) all the >way to top of block. I did not process/embed these tissues. So far I've done >the following: > > 1. changed blades(went threw about 10 just in a few blocks) > 2. cleaned microtome > 3. changed angles(6-12 degree & everything in between) > 4. sectioned hot, cold, freezing > 5. soaked blocks in ice cold water; froze blocks > 6. surfaced decal 5 - 40 minute intervals > 7. re-embedded >The paraffin seems to be causing the lines! I've never experienced problems >like this before. Usually its the tissue itself that causes a nick in blade >which therefore causing vertical lines to appear when sectiong the tissue. >Help is needed! What next is recommended? Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From mohana_g2002 <@t> yahoo.com Tue Jan 31 12:35:44 2006 From: mohana_g2002 <@t> yahoo.com (MOHANA) Date: Tue Jan 31 12:35:53 2006 Subject: [Histonet] neuron Message-ID: <20060131183544.13989.qmail@web33503.mail.mud.yahoo.com> plz can someone send me pictures of different types of neuron stained luxol fast blue with cresyl violet and which is the best magnification . so that it will help me to identify pyramidal, pyramidal like, multipolar and local circuit neurons ! mohana Mohana --------------------------------- Yahoo! Autos. Looking for a sweet ride? Get pricing, reviews, & more on new and used cars. From PIXLEYSK <@t> UCMAIL.UC.EDU Tue Jan 31 12:49:26 2006 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Tue Jan 31 12:49:31 2006 Subject: [Histonet] RE: Brown nickel-DAB Message-ID: Dear Adam: Your explanation seems possible given the pattern of staining with your two antibodies. One thought is that, it seems like your staining is adequate for your purposes without worrying about the color of the staining, so perhaps you could just accept it. But to check it out, I would be tempted to try DAB staining without the nickel enhancement. Or do the nickel-DAB staining with a dilution curve of the antibody. If you dilute your antibody, do you revert to the regular blue-clack color? Another thought is that the brown color means less DAB product, which might occur if the viral vector expression produces a slightly different protein that does not bind as much antibody. Are you expressing the full length receptor, or only part of the receptor? Sarah From sjchtascp <@t> yahoo.com Tue Jan 31 12:55:48 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Tue Jan 31 12:55:56 2006 Subject: [Histonet] processing Message-ID: <20060131185548.50263.qmail@web90210.mail.scd.yahoo.com> Good afternoon, I'm working with mouse/rat muscle and trying to fine tune my processing schecdule. My last run of muscle were very dry. I have a copy of the NSH Animal Processing Manual and trying to strike a balance between those listed on page 5-6. I've noticed most of the protocols do not call for heat or P/V. The scedule I currently use is as follows: 1-70% (hold), 1-80%, 2-95%, 3-100%, 3-xylene all for 30 minutes each and at 38C with P/V. 3 paraffins, 45 minutes each at 55C with P/V. I'm fairly new to research and still trying to make the "mental" switch in handling the different tissue. Thanks everyone, Steve --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. From rgrow <@t> bmnet.com Tue Jan 31 12:57:33 2006 From: rgrow <@t> bmnet.com (rgrow@bmnet.com) Date: Tue Jan 31 12:57:43 2006 Subject: [Histonet] Re: vertical lines in paraffin In-Reply-To: <200601311725.k0VHP93n027023@dns1.bmnet.com> Message-ID: Dina What a coincedence! I had the same problem (years ago) when I worked for Tulane Primate Research Center (now renamed) on the North Shore. If you stain your slides and have no lines in your sections, it may be the silicone coating they put on the slides at the factory. Sometimes it is heavier than normal. Continue to use the same blade and after a couple of blocks, the lines should go away. If you continue to have problems, try stirring your paraffin before embedding. The polymers may not be evenly distributed and will appear as small white "floaters" in your holding tank. Renee Grow, BA., HT (ASCP) rgrow@bmnet.com Histology Supervisor Blount Memorial Hospital 907 E. Lamar Alexander Pkwy. Maryville, TN 37804-5016 (865) 977-4744 (865) 977-5766 Fax From caron_fournier <@t> yahoo.ca Tue Jan 31 12:57:42 2006 From: caron_fournier <@t> yahoo.ca (caron fournier) Date: Tue Jan 31 12:57:51 2006 Subject: [Histonet] re: bone section problem Message-ID: <20060131185743.70827.qmail@web35406.mail.mud.yahoo.com> Hi: anyone out there doing bone sectioning and grinding. I am having a problem with some of the sections not taking up stain in certain areas but another sample on the same section works fine. Staining is done the same and yet they are not uniform. Has anyone had any problems with the Technovit glue coming through into the section and affecting the staining after they are ground down to 25 microns? The stain being used is for the calcified bone and areas look like someone has taken an eraser and erased the stain along lines in the cortical bone. Also, on some sections the edges of the section are lifting and the toluidine blue stain is bleeding under the section and into the glue. Any suggestions? Thank you, Caron --------------------------------- Find your next car at Yahoo! Canada Autos From Jeffery-Smith <@t> ouhsc.edu Tue Jan 31 13:17:56 2006 From: Jeffery-Smith <@t> ouhsc.edu (Smith, Jeffery D. (HSC)) Date: Tue Jan 31 13:21:50 2006 Subject: [Histonet] processing Message-ID: Sounds to me like too much 100% alcohol. Try using 2 80% and only 2 100% this should eliminate the overhardening yet still give you plenty of dehydration. The next suggestion at that temp. with P/V try cutting back the times in 100% and add some time to the lower grade alcohols. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Steven Coakley Sent: Tue 1/31/2006 12:55 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] processing Good afternoon, I'm working with mouse/rat muscle and trying to fine tune my processing schecdule. My last run of muscle were very dry. I have a copy of the NSH Animal Processing Manual and trying to strike a balance between those listed on page 5-6. I've noticed most of the protocols do not call for heat or P/V. The scedule I currently use is as follows: 1-70% (hold), 1-80%, 2-95%, 3-100%, 3-xylene all for 30 minutes each and at 38C with P/V. 3 paraffins, 45 minutes each at 55C with P/V. I'm fairly new to research and still trying to make the "mental" switch in handling the different tissue. Thanks everyone, Steve --------------------------------- Bring words and photos together (easily) with PhotoMail - it's free and works with Yahoo! Mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sandra.Harrison3 <@t> va.gov Tue Jan 31 13:27:58 2006 From: Sandra.Harrison3 <@t> va.gov (Harrison, Sandra C.) Date: Tue Jan 31 13:28:09 2006 Subject: [Histonet] cleaning a cryostat after possible contamination with t.b. Message-ID: <736E8889E98B8F4FBBD29FDFEC8074BA49BFAF@VHAV23MSGA2.v23.med.va.gov> Dear Histonetters, I've checked with numerous other labs, also with CAP, to try to determine the best method for decontaminating a cryostat after a specimen has been cut on it that might be t.b. Almost everyone agrees that the cryostat needs to be shut down, brought to room temp, and exposed to a disinfectant. The variations seem to be in which disinfectant. Some use 37% formalin in an open container placed in the cryostat, allowing it to remain in there overnight so that the cryostat is exposed to the fumes. Then the interior is wiped down with absolute alcohol. After 24-48 hours, to ensure complete dryness, the cryostat is turned back on. Some people use a liquid phenolic, rather than formalin fumes and then absolute alcohol. The downside of using the formalin fumes is that some have reported pitting in the cryostats interior. Of course, the best of all would be to have one of the self de-contaminating cryostats which use either ultra-violet light or have a well for the 37% formalin and is an automatic cycle. Please tell me what methods YOU are finding best and why. Thanks, Sandy From Jan.Minshew <@t> leica-microsystems.com Tue Jan 31 14:01:52 2006 From: Jan.Minshew <@t> leica-microsystems.com (Jan.Minshew@leica-microsystems.com) Date: Tue Jan 31 14:01:56 2006 Subject: [Histonet] cleaning a cryostat after possible contamination with t.b. In-Reply-To: <736E8889E98B8F4FBBD29FDFEC8074BA49BFAF@VHAV23MSGA2.v23.med.va.gov> Message-ID: Hi Sandy, I posted this several months ago. I hope it helps you establish a protocol. If you have additional questions, please feel free to contact me. I'll be happy to help. Don't forget, when you have completed the disinfection protocol you must completely dry the chamber, microtome and all accessories. The instrument should be lubricated according to the manufacturers specifications, using the recommended lubricant, if it is going to be put back into service at cold temperatures. >> Whether a cryostat has a built-in disinfection system or not, there are several very important things to remember about disinfecting cryostats. 1. Before beginning a disinfection protocol, don personal protective equipment (puncture and penetration resistant gloves, gowns, etc). 2. Remove all debris from the cryostat and disposed of it according to the policies and procedures of your institution. The debris must be removed because organic material (blood and proteins) may contain high concentrations of microorganisms and could possibly inactivate chemical germicides or prevent access to contaminated surfaces. 3. Use 70% ethyl (or reagent) alcohol to clean the cryostat. The germicidal activity of ethyl alcohol is most effective in that range and it has an advantage over isopropyl alcohol of being able to kill hydrophilic viruses. 4. For those of us in the USA (other countries have access to other products), the EPA maintains a list of Antimicrobial Chemical/Registration Number Indexes and it is posted on their website http://www.epa.gov/oppad001/chemregindex.htm. From this link you can find agents effective against bloodborne pathogens such as Mycobacterium tuberculosis, human HIV-1 virus, Hepatitis B or Hepatitis C virus. It is critical to remember that NONE of these solutions have been tested at low temperatures and they can only be used at room temperature. 5. Do not create aerosols by spraying disinfectant (or anything else) in an open cryostat chamber. Pour disinfectants onto absorbent disposable towels and allow them to remain in contact with contaminated surfaces for the length of time specified in the instructions of the individual agents. << I hope this information is useful. Please let me know if you have any questions. Best wishes to all, Jan Minshew HT, HTL(ASCP) Marketing Manager Leica Microsystems, Inc. 2345 Waukegan Rd Bannockburn, IL 60015 800.248.0123 x7051 "Harrison, Sandra C." Sent by: cc histonet-bounces@ lists.utsouthwest Subject ern.edu [Histonet] cleaning a cryostat after possible contamination with t.b. 01/31/2006 01:27 PM Dear Histonetters, I've checked with numerous other labs, also with CAP, to try to determine the best method for decontaminating a cryostat after a specimen has been cut on it that might be t.b. Almost everyone agrees that the cryostat needs to be shut down, brought to room temp, and exposed to a disinfectant. The variations seem to be in which disinfectant. Some use 37% formalin in an open container placed in the cryostat, allowing it to remain in there overnight so that the cryostat is exposed to the fumes. Then the interior is wiped down with absolute alcohol. After 24-48 hours, to ensure complete dryness, the cryostat is turned back on. Some people use a liquid phenolic, rather than formalin fumes and then absolute alcohol. The downside of using the formalin fumes is that some have reported pitting in the cryostats interior. Of course, the best of all would be to have one of the self de-contaminating cryostats which use either ultra-violet light or have a well for the 37% formalin and is an automatic cycle. Please tell me what methods YOU are finding best and why. Thanks, Sandy _______________________________________________ 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 JMahoney <@t> alegent.org Tue Jan 31 14:12:36 2006 From: JMahoney <@t> alegent.org (Janice Mahoney) Date: Tue Jan 31 14:13:26 2006 Subject: [Histonet] cleaning a cryostat after possible contamination witht.b. Message-ID: The only thing I have found to be non-corrosive is the alcohol or UV light. All the things suggested will kill TB Jan Omaha >>> "Harrison, Sandra C." 01/31/2006 1:27 PM >>> Dear Histonetters, I've checked with numerous other labs, also with CAP, to try to determine the best method for decontaminating a cryostat after a specimen has been cut on it that might be t.b. Almost everyone agrees that the cryostat needs to be shut down, brought to room temp, and exposed to a disinfectant. The variations seem to be in which disinfectant. Some use 37% formalin in an open container placed in the cryostat, allowing it to remain in there overnight so that the cryostat is exposed to the fumes. Then the interior is wiped down with absolute alcohol. After 24-48 hours, to ensure complete dryness, the cryostat is turned back on. Some people use a liquid phenolic, rather than formalin fumes and then absolute alcohol. The downside of using the formalin fumes is that some have reported pitting in the cryostats interior. Of course, the best of all would be to have one of the self de-contaminating cryostats which use either ultra-violet light or have a well for the 37% formalin and is an automatic cycle. Please tell me what methods YOU are finding best and why. Thanks, Sandy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Tue Jan 31 14:16:22 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Tue Jan 31 14:16:37 2006 Subject: [Histonet] cleaning a cryostat after possible contamination with t.b. Message-ID: I shut it down and bring to room temp, then I use Sanimster IV, which is a broad spectrum killer of all, then wipe it out with 100%, DONE! Turn on and good to go. Robyn OHSU From tkngflght <@t> yahoo.com Tue Jan 31 14:56:29 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Tue Jan 31 14:56:38 2006 Subject: [Histonet] Trying to find old histo friends, please help? Message-ID: <20060131205629.44716.qmail@web50906.mail.yahoo.com> Hi Histonetters-- I am trying to find old histology friends. I was a traveler for a number of years and have lived and worked all over. If you ever worked with me, know me from social gatherings or THINK you might know me, please drop me a line? I'm looking for some of you specifically and would love to reconnect with all of you--you all taught me so much and I wish I'd done a better job of keeping up!! Looking forward to hearing from you! Cheryl Kerry, HT(ASCP) tkngflght@yahoo.com 281.883.7704 From KDwyer3322 <@t> aol.com Tue Jan 31 17:06:50 2006 From: KDwyer3322 <@t> aol.com (KDwyer3322@aol.com) Date: Tue Jan 31 17:07:02 2006 Subject: [Histonet] Texas Society for Histotechnolgy 2006 Symposium/Convention Message-ID: <36.2afe3f3.3111478a@aol.com> To all, The Texas Society for Histotechnology 2006 Convention will be held March 31-April 2, 2006 at the Omni Hotel in Corpus Christi, Texas. For more information and program please go to our web site at Txsh.org. or contact Judy Webb at 817-927-1024 or Veronica Davis at 972-579-8291. Programs will be mailed February 1, 2006. Deadline for hotel reservations is March 9, 2006. Hope to see you there.