From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 1 02:18:41 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Re: Ammonia water pH and other bluing solutions[Sc anned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B54C@bhrv-nt-11.bhrv.nwest.nhs.uk> I think you are wrong, so I'll correct you, as you requested. I thought the 'blue' state of haematin was, like litmus, the effect an alkaline condition has on the siderphilic dye. Red, like litmus, denoted acidic conditions. Damned if I know what colour (with the 'u') haematin goes when neutral (bluey/ red?) Running tap water takes longer, I concede, but good things are worth waiting for and it's easier to make up; we rush too much I find. TIP: Never use London water, been through too many kidneys! -----Original Message----- From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: 30 November 2004 19:09 To: Johnson, Teri Cc: Histonet; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions[Scanned] Anyone remember using saturated lithium carbonate? I could probably use a little lithium right now . . . . . Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just bringing the slides BACK to a neutral pH after treating them with acid which makes them purple-ish? I like the ammonia because it 'seems' to make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to make them "blah".(Perception is in the eye of the beholder) Running tap water will make the same miracle happen, but it takes longer. "Johnson, Teri" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2004 12:51 PM To: "Histonet" cc: Subject: [Histonet] Re: Ammonia water pH and other bluing solutions Way back in the day, Brigati was using PBS on his immunostainer to blue after the hematoxylin counterstain. I have since used PBS to blue my slides instead of ammonium hydroxide/water. It's cheap, and very easy on the sections, and I have plenty of it on hand. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 1 02:25:23 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] gliut/isopropyl alcohol[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B54D@bhrv-nt-11.bhrv.nwest.nhs.uk> Expect the tissue is rather brittle, used to be used for electromicroscopy I think. Wonder if you could secondary fix? Can't aldehyde/ alcohol fixation be 'washed out' then the tissue refixed in something more conventional? -----Original Message----- From: Molinari, Betsy [mailto:BMolinari@heart.thi.tmc.edu] Sent: 30 November 2004 19:08 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] gliut/isopropyl alcohol[Scanned] Hi histonetters, I received samples that have been fixed in a 1%glut / 20% isopropyl alcohol solution for paraffin processing. I have never heard of this fixative, I know alcoholic formalin but never alcoholic glutaraldehyde. Are any of you familiar with this fixative? Thank you. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BMolinari <@t> heart.thi.tmc.edu Wed Dec 1 06:18:48 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] gliut/isopropyl alcohol Message-ID: Hi Bob, Thanks for your input. I may ask a couple of the surgery staff here if they are familiar with that technique. A point of interest, these are valves. I loaded them last night starting in 70% ETOH, I am curious to see how they turned out and how much effect it will have on the B&B and PTAH I will be running on them Thanks again, Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology Houston, TX 77030 832-355-6524 -----Original Message----- From: RCHIOVETTI@aol.com [mailto:RCHIOVETTI@aol.com] Sent: Tuesday, November 30, 2004 6:26 PM To: Molinari, Betsy; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] gliut/isopropyl alcohol In a message dated 11/30/2004 12:12:08 PM US Mountain Standard Time, BMolinari@heart.thi.tmc.edu writes: I received samples that have been fixed in a 1%glut / 20% isopropyl alcohol solution for paraffin processing. I have never heard of this fixative, I know alcoholic formalin but never alcoholic glutaraldehyde. Are any of you familiar with this fixative? Thank you. Betsy Molinari HT (ASCP) Hi Betsy, Is the specimen perhaps a heart valve made from pig tissue?? The only time I've heard of alcoholic glutaraldehyde is for pretreating pig valves before they're implanted in humans. There's some kind of relationship between alcohol and glut, and preventing calcification of the valve. Can't remember the details right now, but you have to treat the valve with one of those reagents before using the other. Just a thought... Cheers, Bob Chiovetti From BMolinari <@t> heart.thi.tmc.edu Wed Dec 1 06:26:11 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] gliut/isopropyl alcohol[Scanned] Message-ID: Hi Kemlo, Thanks for your suggestion, I did not think of rinsing! The EM tech here had never heard of a glut/isopropyl mix, we use a 2%glut solution in buffer for the EM done here. I will be cutting and staining the blocks today, it will be interesting. Thanks, Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston, TX 77373 832-355-6524 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kemlo Rogerson Sent: Wednesday, December 01, 2004 2:25 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] gliut/isopropyl alcohol[Scanned] Expect the tissue is rather brittle, used to be used for electromicroscopy I think. Wonder if you could secondary fix? Can't aldehyde/ alcohol fixation be 'washed out' then the tissue refixed in something more conventional? -----Original Message----- From: Molinari, Betsy [mailto:BMolinari@heart.thi.tmc.edu] Sent: 30 November 2004 19:08 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] gliut/isopropyl alcohol[Scanned] Hi histonetters, I received samples that have been fixed in a 1%glut / 20% isopropyl alcohol solution for paraffin processing. I have never heard of this fixative, I know alcoholic formalin but never alcoholic glutaraldehyde. Are any of you familiar with this fixative? Thank you. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 _______________________________________________ Histonet mailing list Histonet@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 NHeath <@t> Lifespan.org Wed Dec 1 07:40:38 2004 From: NHeath <@t> Lifespan.org (Heath, Nancy L.) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods? Message-ID: <09C945920A6B654199F7A58A1D7D1FDE65EC58@lsexch.lsmaster.lifespan.org> HI Richard, There are new regs. in place for shipping. Yes when something is considered "dangerous goods" it has to be packaged & shipped by trained personnel. SafTPak puts out a great cd for this training. You can go to there website and pick one up. Nancy -----Original Message----- From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: Tuesday, November 30, 2004 12:14 PM To: Richard Cartun Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Paraffin blocks - Dangerous goods? Unfortunately, they are considered a biohazard -(human tissue), although my sister unknowingly sent our dead aunt's remains (ashes) via UPS once. I came home from work and she was right there in a little box, sitting on my front porch. Anyway, I digress - I remember this being discussed before as to why they (blocks) are still biohazardous since they're fixed and processed. I'm facing storage problems right now because we don't have anywhere 'appropriate' to keep them. Years (and years) ago - we stored them in the basements and sub-basements (scary 4 foot ceilings) of hospitals until they were found by lucky rodents or other vermin. I'm curious as to how and where people store them now, as well. Jackie "Richard Cartun" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2004 10:12 AM To: cc: Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tissuearray <@t> hotmail.com Wed Dec 1 07:58:13 2004 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Microarrayer In-Reply-To: <6a.497e6d96.2edd1c05@aol.com> Message-ID: From kgrobert <@t> rci.rutgers.edu Wed Dec 1 08:35:15 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Re: Ammonia water pH and other bluing solutions In-Reply-To: References: Message-ID: <41ADD6A3.6070901@rci.rutgers.edu> Heck, we still use that. All our slides look lovely. The mellow one- Kathleen Roberts Rutgers University Jackie M O'Connor wrote: >Anyone remember using saturated lithium carbonate? I could probably use a >little lithium right now . . . . . > >Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just >bringing the slides BACK to a neutral pH after treating them with acid >which makes them purple-ish? I like the ammonia because it 'seems' to >make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to >make them "blah".(Perception is in the eye of the beholder) Running tap >water will make the same miracle happen, but it takes longer. > > > > > > >"Johnson, Teri" >Sent by: histonet-bounces@lists.utsouthwestern.edu >11/30/2004 12:51 PM > > > To: "Histonet" > cc: > Subject: [Histonet] Re: Ammonia water pH and other bluing solutions > > >Way back in the day, Brigati was using PBS on his immunostainer to blue >after the hematoxylin counterstain. I have since used PBS to blue my >slides instead of ammonium hydroxide/water. It's cheap, and very easy >on the sections, and I have plenty of it on hand. > > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, Missouri 64110 >tjj@stowers-institute.org > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 1 08:37:56 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Re: Ammonia water pH and other bluing solutions[Sc anned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B55B@bhrv-nt-11.bhrv.nwest.nhs.uk> Nice and red? -----Original Message----- From: Kathleen Roberts [mailto:kgrobert@rci.rutgers.edu] Sent: 01 December 2004 14:35 To: Jackie M O'Connor; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions[Scanned] Heck, we still use that. All our slides look lovely. The mellow one- Kathleen Roberts Rutgers University Jackie M O'Connor wrote: >Anyone remember using saturated lithium carbonate? I could probably use a >little lithium right now . . . . . > >Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just >bringing the slides BACK to a neutral pH after treating them with acid >which makes them purple-ish? I like the ammonia because it 'seems' to >make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to >make them "blah".(Perception is in the eye of the beholder) Running tap >water will make the same miracle happen, but it takes longer. > > > > > > >"Johnson, Teri" >Sent by: histonet-bounces@lists.utsouthwestern.edu >11/30/2004 12:51 PM > > > To: "Histonet" > cc: > Subject: [Histonet] Re: Ammonia water pH and other bluing solutions > > >Way back in the day, Brigati was using PBS on his immunostainer to blue >after the hematoxylin counterstain. I have since used PBS to blue my >slides instead of ammonium hydroxide/water. It's cheap, and very easy >on the sections, and I have plenty of it on hand. > > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, Missouri 64110 >tjj@stowers-institute.org > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@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 lwhite <@t> lakeridgehealth.on.ca Wed Dec 1 08:47:19 2004 From: lwhite <@t> lakeridgehealth.on.ca (White, Lori ) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] RE: Autoimmunostainers DAKO vs Ventanna Message-ID: I have used both systems and there are pros and cons to each. It really is dependent on your IHC volume and how you want to use the automation. If you want to be free of working up antibodies from concentrates to maximize dilution and are willing to pay the expense of prediluted reagents, the Ventana system is probably the better way to go. Keep in mind that you are locked into one source for all reagents including buffers and detection. To me, this is a disadvantage. If you have a fairly high volume IHC laboratory and experienced staff, I would recommend the open systems. You have a lot more flexibility for not only primary antibodies but all of your ancillary reagents as well. One of the features I liked about the Dako was that you were able to see what was happening during the staining run, not only on the instrument but on the computer screen as well. I also liked the sensor/alarm for dispensing of reagents. In terms of service, I would say that again both systems have positives and negatives. My two cents...... Lori White Charge Technologist Lakeridge Health Corporation Oshawa, Ontario Canada -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Wednesday, December 01, 2004 7:26 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 13, Issue 1 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. Ammonia water pH (Paula Pierce) 2. Frozen cell pellet (Susan Q Wells) 3. RE: Frozen cell pellet (Favara, Cynthia (NIH/NIAID)) 4. Re: Ammonia water pH and other bluing solutions (Johnson, Teri) 5. gliut/isopropyl alcohol (Molinari, Betsy) 6. Re: Re: Ammonia water pH and other bluing solutions (Jackie M O'Connor) 7. Re: Re: Ammonia water pH and other bluing solutions (Robyn Vazquez) 8. RE: Autoimmunostainers DAKO vs Ventanna[Scanned] (Darren James) 9. Re: Frozen cell pellet (Gayle Callis) 10. Saturated lithium carbonate bluing (Gayle Callis) 11. RE: Autoimmunostainers DAKO vs Ventanna[Scanned] (Darren James) 12. Re: Ammonia water pH and other bluing solutions (Johnson, Teri) 13. Paraffin processing explant cultures (Johnson, Teri) 14. Re: Re: Ammonia water pH and other bluing solutions (Gudrun Lang) 15. RE: Paraffin blocks - Dangerous goods? (Luck, Greg D.) 16. RE: Counterstain for Von Kossa (Fredrik Johansson) 17. Phosphorylated-met antibodies (Tan, MinHan) 18. RE: Paraffin processing explant cultures (Elizabeth Chlipala) 19. CD 133 (nperson211@comcast.net) 20. RE: Paraffin processing explant cultures (Johnson, Teri) 21. RE: Paraffin blocks - Dangerous goods? (Robyn Vazquez) 22. Re: Ammonia water pH (SMITH,REBEKAH FELICIA) 23. Re: Paraffin blocks - Dangerous goods? (Jennifer MacDonald) 24. Re: gliut/isopropyl alcohol (RCHIOVETTI@aol.com) 25. Part time Histology position in San Diego Ca. (James Watson) 26. RE: Re: Ammonia water pH and other bluing solutions[Sc anned] (Kemlo Rogerson) 27. RE: gliut/isopropyl alcohol[Scanned] (Kemlo Rogerson) 28. RE: gliut/isopropyl alcohol (Molinari, Betsy) ---------------------------------------------------------------------- Message: 1 Date: Tue, 30 Nov 2004 10:17:12 -0800 (PST) From: Paula Pierce Subject: [Histonet] Ammonia water pH To: histonet@lists.utsouthwestern.edu Message-ID: <20041130181712.32329.qmail@web50303.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii lol, Jackie O'Conner. I like you. I do the same thing. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com ------------------------------ Message: 2 Date: Tue, 30 Nov 2004 13:21:51 -0500 From: Susan Q Wells Subject: [Histonet] Frozen cell pellet To: Histonet@lists.utsouthwestern.edu Message-ID: <41ACBA3F.9030602@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Does anyone have advise on how to freeze cells that have been spun down in a micro centrifuge tube? In the past I've snap frozen the tube, loosened the cells by flicking the tube and then embedded the pellet in OCT. The cells look OK, but the cells in the middle don't look as good as the ones on the perimeter.Any advise would be appreciated. Thanks, Sue Wells ------------------------------ Message: 3 Date: Tue, 30 Nov 2004 13:38:40 -0500 From: "Favara, Cynthia (NIH/NIAID)" Subject: RE: [Histonet] Frozen cell pellet To: 'Susan Q Wells' , Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain I have done as you have, also spin down add a bit of histogel vortex let cool then flick out and put in OCT. I like this as I seem to get a more even distribution of cells. 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: Susan Q Wells [mailto:susan.wells@bms.com] Sent: Tuesday, November 30, 2004 11:22 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Frozen cell pellet Does anyone have advise on how to freeze cells that have been spun down in a micro centrifuge tube? In the past I've snap frozen the tube, loosened the cells by flicking the tube and then embedded the pellet in OCT. The cells look OK, but the cells in the middle don't look as good as the ones on the perimeter.Any advise would be appreciated. Thanks, Sue Wells _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Tue, 30 Nov 2004 12:51:22 -0600 From: "Johnson, Teri" Subject: [Histonet] Re: Ammonia water pH and other bluing solutions To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" Way back in the day, Brigati was using PBS on his immunostainer to blue after the hematoxylin counterstain. I have since used PBS to blue my slides instead of ammonium hydroxide/water. It's cheap, and very easy on the sections, and I have plenty of it on hand. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org ------------------------------ Message: 5 Date: Tue, 30 Nov 2004 13:07:43 -0600 From: "Molinari, Betsy" Subject: [Histonet] gliut/isopropyl alcohol To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi histonetters, I received samples that have been fixed in a 1%glut / 20% isopropyl alcohol solution for paraffin processing. I have never heard of this fixative, I know alcoholic formalin but never alcoholic glutaraldehyde. Are any of you familiar with this fixative? Thank you. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 ------------------------------ Message: 6 Date: Tue, 30 Nov 2004 13:08:41 -0600 From: "Jackie M O'Connor" Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions To: "Johnson, Teri" Cc: Histonet , histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Anyone remember using saturated lithium carbonate? I could probably use a little lithium right now . . . . . Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just bringing the slides BACK to a neutral pH after treating them with acid which makes them purple-ish? I like the ammonia because it 'seems' to make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to make them "blah".(Perception is in the eye of the beholder) Running tap water will make the same miracle happen, but it takes longer. "Johnson, Teri" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2004 12:51 PM To: "Histonet" cc: Subject: [Histonet] Re: Ammonia water pH and other bluing solutions Way back in the day, Brigati was using PBS on his immunostainer to blue after the hematoxylin counterstain. I have since used PBS to blue my slides instead of ammonium hydroxide/water. It's cheap, and very easy on the sections, and I have plenty of it on hand. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Tue, 30 Nov 2004 11:38:08 -0800 From: "Robyn Vazquez" Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions To: Jackie.O'Connor@abbott.com, histonet-bounces@lists.utsouthwestern.edu, TJJ@Stowers-Institute.org Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Jackie, I use a saturated lithium carbonate for my frozen Mohs tissue. Works and looks great... Robyn OHSU >>> "Jackie M O'Connor" 11/30/2004 11:08:41 AM >>> Anyone remember using saturated lithium carbonate? I could probably use a little lithium right now . . . . . Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just bringing the slides BACK to a neutral pH after treating them with acid which makes them purple-ish? I like the ammonia because it 'seems' to make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to make them "blah".(Perception is in the eye of the beholder) Running tap water will make the same miracle happen, but it takes longer. "Johnson, Teri" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2004 12:51 PM To: "Histonet" cc: Subject: [Histonet] Re: Ammonia water pH and other bluing solutions Way back in the day, Brigati was using PBS on his immunostainer to blue after the hematoxylin counterstain. I have since used PBS to blue my slides instead of ammonium hydroxide/water. It's cheap, and very easy on the sections, and I have plenty of it on hand. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Wed, 1 Dec 2004 08:49:29 +1300 From: "Darren James" Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] To: "'Kemlo Rogerson'" , "Histonet \(E-mail\)" Message-ID: <000001c4d715$b49168d0$c864a8c0@medica.co.nz> Content-Type: text/plain; charset="iso-8859-1" As another option take a look at the Bond Max manufactured by Vision Bio Systems http://www.vision-bio.com/product_bondmax.html It performs on board dewaxing, heat induced epitope retreival, enzyme digestion as well as counterstaining. I have only heard good things about it. I have used the Dako extensively and although good, it has it's limitations. At the time we chose the Dako over the Ventana due to the Ventana being a closed system. If I were in a position to purchase another I would definitely take a long hard look at the Bond Max. Thanks and have a great day Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kemlo Rogerson Sent: Wednesday, 1 December 2004 4:36 a.m. To: histonet@pathology.swmed.edu Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] WE have both, and I'm told the Ventana is the 'better' of the two options. As an ICC virgin and one who will have to choose which platform to go with I am particularly interested in this thread. I would prefer to use the cheapest 'open' system that is a automated as possible, is that mutually exclusive? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Maliniak, Richard [mailto:RMaliniak@SBHCS.com] Sent: 30 November 2004 15:16 To: histonet@pathology.swmed.edu Subject: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] I am looking for info= rmation from people who have switched from Ventanna to DAKO and DAKO to V= entanna. Why did you switch? What did you like or dislike abo= ut one or the other. Rick ________________________________________________________________ Important news about our email communications Saint Barnabas Health Care System has implemented secure messaging servic= es. To learn more about SBHCS Secure Messaging, go to: [1]http://www.zixcorp.c= om/evangelism/sbhcs/ If you need assistance with retrieving a secure email, please email sbhcs= accounts@sbhcs.com or visit [2]http://www.zixcorp.com/evangelism/sbhcs/par= tners/receiving.php References 1. 3D"http://www.zixcorp.com/evangelism/sbhcs/" 2. 3D"http://www.zixcorp.com/evangelism/_______________________________________ ________ Histonet mailing list Histonet@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: 9 Date: Tue, 30 Nov 2004 12:45:41 -0700 From: Gayle Callis Subject: Re: [Histonet] Frozen cell pellet To: Susan Q Wells , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041130123759.01b57008@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed How many cells you spin down may be critical. Here is a protocol that works for us. 1. 1 to 3 x 10 (to the 7th) cells/ml. 2. Spin down and rinse three time with PBS. 3. Add OCT (approx 1 ml or less) to last pellet, mix. 4. Immerse tube directly into liquid N2. 5. Pop block out with a sharp rap inside cryostat. Mount block and section. The key is to make sure you don't have too many cells and do a good mix with OCT in order to disperse any water from buffer and spread out cells. We have sectioned thinner, 4 um rather than 5 to insure uncrowded staining. We build block up with more OCT around end of block so the block face is not so tiny and you can manipulate the tiny section. Chris van der Loos is a master at the supertiny section!! A wider conical end microcentrifuge tube (holds 2 ml or more overall) spreads things out a bit more for a broader faced block but a 1.5 ml tube will work. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 10 Date: Tue, 30 Nov 2004 12:58:26 -0700 From: Gayle Callis Subject: [Histonet] Saturated lithium carbonate bluing To: "Jackie M O'Connor" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041130124819.01b66e58@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Yes, have used sat LiCO3, but it had to be decanted carefully into the container or the lithium carbonate crystals poured off polarized!! Bummer with congo red amyloid staining to see annoying crystals everywhere. And then the saturated stuff grew some ugly black floating grow-ty stuff, a touch of black slime. Went back to Scotts tap water substitute or Richard Allen bluing solution. An excellent article on the theory of hematoxylin staining is found in J of Histotechnology, Susan Meloan and H. Puchtler volume 10, 1987. The actual staining by hematoxylin, nomenclature, and what the dye molecule consists of, along with what acid and alkaline solutions does is explained in great detail. After reading this publication, how I do or think about hematoxylin staining (either progressive or regressive methods) was radically changed. It is very chemical, but very informative - be prepared for some interesting chemistry. I strongly recommend this publication to all doing H&E staining. At 12:08 PM 11/30/2004, you wrote: >Anyone remember using saturated lithium carbonate? I could probably use a >little lithium right now . . . . . > >Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just >bringing the slides BACK to a neutral pH after treating them with acid >which makes them purple-ish? I like the ammonia because it 'seems' to >make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to >make them "blah".(Perception is in the eye of the beholder) Running tap >water will make the same miracle happen, but it takes longer. > > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 11 Date: Wed, 1 Dec 2004 09:28:05 +1300 From: "Darren James" Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] To: "'Bonnie Whitaker'" , "Histonet \(E-mail\)" Message-ID: <000801c4d71b$198aeb80$c864a8c0@medica.co.nz> Content-Type: text/plain; charset="us-ascii" Hi Bonnie, I don't know about the US and the UK but here in NZ and also Australia they are being placed at various sites. I would assume that if there were any pending legal action the hold on selling would be global. Thanks Darren -----Original Message----- From: Bonnie Whitaker [mailto:bwhitaker@brownpathology.com] Sent: Wednesday, 1 December 2004 9:01 a.m. To: darrenj@medica.co.nz Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] I heard that they were held up in litigation, and would be unable to sell the instruments for at least a year..... Also, it's a closed system (at least to some degree.) Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Darren James Sent: Tuesday, November 30, 2004 1:49 PM To: 'Kemlo Rogerson'; Histonet (E-mail) Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] As another option take a look at the Bond Max manufactured by Vision Bio Systems http://www.vision-bio.com/product_bondmax.html It performs on board dewaxing, heat induced epitope retreival, enzyme digestion as well as counterstaining. I have only heard good things about it. I have used the Dako extensively and although good, it has it's limitations. At the time we chose the Dako over the Ventana due to the Ventana being a closed system. If I were in a position to purchase another I would definitely take a long hard look at the Bond Max. Thanks and have a great day Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kemlo Rogerson Sent: Wednesday, 1 December 2004 4:36 a.m. To: histonet@pathology.swmed.edu Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] WE have both, and I'm told the Ventana is the 'better' of the two options. As an ICC virgin and one who will have to choose which platform to go with I am particularly interested in this thread. I would prefer to use the cheapest 'open' system that is a automated as possible, is that mutually exclusive? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Maliniak, Richard [mailto:RMaliniak@SBHCS.com] Sent: 30 November 2004 15:16 To: histonet@pathology.swmed.edu Subject: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] I am looking for info= rmation from people who have switched from Ventanna to DAKO and DAKO to V= entanna. Why did you switch? What did you like or dislike abo= ut one or the other. Rick ________________________________________________________________ Important news about our email communications Saint Barnabas Health Care System has implemented secure messaging servic= es. To learn more about SBHCS Secure Messaging, go to: [1]http://www.zixcorp.c= om/evangelism/sbhcs/ If you need assistance with retrieving a secure email, please email sbhcs= accounts@sbhcs.com or visit [2]http://www.zixcorp.com/evangelism/sbhcs/par= tners/receiving.php References 1. 3D"http://www.zixcorp.com/evangelism/sbhcs/" 2. 3D"http://www.zixcorp.com/evangelism/_______________________________________ ________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Tue, 30 Nov 2004 15:09:05 -0600 From: "Johnson, Teri" Subject: [Histonet] Re: Ammonia water pH and other bluing solutions To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" FYI, the pH of the PBS we use is 7.2 but I'm sure any neutral to slightly alkaline pH would work fine. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org ------------------------------ Message: 13 Date: Tue, 30 Nov 2004 15:13:43 -0600 From: "Johnson, Teri" Subject: [Histonet] Paraffin processing explant cultures To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" Is anybody doing paraffin processing on explant cultures and can give me some tips on increasing my chances for success? The culture plate inserts are Millicell polycarbonate and my question is whether they can survive tissue processing and paraffin sectioning. If not, what are your recommendations for doing these? Thanks in advance! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org ------------------------------ Message: 14 Date: Tue, 30 Nov 2004 22:59:33 +0100 From: "Gudrun Lang" Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions To: "Histonetliste" Message-ID: <003a01c4d727$e077a3d0$eeeea8c0@server> Content-Type: text/plain; charset="iso-8859-1" Using lithiumcarbonat is the way we do it with staining frozen sections. In the HE-stainer we have running tapwater. And the sections are nice. After haemalaun in special stains we let the slides in tapwater for 3-5 min. Gudrun Lang ----- Original Message ----- From: "Jackie M O'Connor" To: "Johnson, Teri" Cc: "Histonet" ; Sent: Tuesday, November 30, 2004 8:08 PM Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions > Anyone remember using saturated lithium carbonate? I could probably use a > little lithium right now . . . . . > > Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just > bringing the slides BACK to a neutral pH after treating them with acid > which makes them purple-ish? I like the ammonia because it 'seems' to > make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to > make them "blah".(Perception is in the eye of the beholder) Running tap > water will make the same miracle happen, but it takes longer. > > > > > > > "Johnson, Teri" > Sent by: histonet-bounces@lists.utsouthwestern.edu > 11/30/2004 12:51 PM > > > To: "Histonet" > cc: > Subject: [Histonet] Re: Ammonia water pH and other bluing solutions > > > Way back in the day, Brigati was using PBS on his immunostainer to blue > after the hematoxylin counterstain. I have since used PBS to blue my > slides instead of ammonium hydroxide/water. It's cheap, and very easy > on the sections, and I have plenty of it on hand. > > > Teri Johnson > Managing Director Histology Facility > Stowers Institute for Medical Research > 1000 E. 50th St. > Kansas City, Missouri 64110 > tjj@stowers-institute.org > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 15 Date: Tue, 30 Nov 2004 13:59:19 -0800 From: "Luck, Greg D." Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? To: 'Richard Cartun' , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Richard, Would you further define or point me towards a regulatory definition of "Dangerous Goods". Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 8:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Tue, 30 Nov 2004 13:59:42 -0800 From: Fredrik Johansson Subject: RE: [Histonet] Counterstain for Von Kossa To: histonet@lists.utsouthwestern.edu Message-ID: <239E252E-431B-11D9-94FE-000A95F0ED90@u.washington.edu> Content-Type: text/plain; charset=WINDOWS-1252; format=flowed Hi ? Thanks for all the answers about counterstain with Von Kossa. ? It seemed like most of you preferred nuclear fast red. So I'm going to try that. ? Thanks ? Fredrik Johansson Dept. of Pathology UW, Seattle ------------------------------ Message: 17 Date: Tue, 30 Nov 2004 17:02:35 -0500 From: "Tan, MinHan" Subject: [Histonet] Phosphorylated-met antibodies To: Message-ID: Content-Type: text/plain; charset="us-ascii" Good afternoon, We are thinking of staining for phosphorylated-met; while there are a number of commercial antibodies out there, it seems that they have not been tested out on paraffin embedded tissue yet. If anyone has tried staining for this antigen, I'd appreciate any pointers you have. Min-Han This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient(s) please contact the sender by reply email and destroy all copies of the original message. Thank you. ------------------------------ Message: 18 Date: Tue, 30 Nov 2004 15:02:30 -0700 From: "Elizabeth Chlipala" Subject: RE: [Histonet] Paraffin processing explant cultures To: "'Johnson, Teri'" , "'Histonet'" Message-ID: <000001c4d728$49c9c480$76d48a80@AMY> Content-Type: text/plain; charset="us-ascii" Teri I have processed explant cultures with the Millipore MillicellT CM which is a polytetrafluoroethylene based polymer. I process these as any other tissue. Millipore has a great web page in which you can ask questions. I would give them a call about the particular filter that you are using. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Tuesday, November 30, 2004 2:14 PM To: Histonet Subject: [Histonet] Paraffin processing explant cultures Is anybody doing paraffin processing on explant cultures and can give me some tips on increasing my chances for success? The culture plate inserts are Millicell polycarbonate and my question is whether they can survive tissue processing and paraffin sectioning. If not, what are your recommendations for doing these? Thanks in advance! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Tue, 30 Nov 2004 22:24:50 +0000 From: nperson211@comcast.net Subject: [Histonet] CD 133 To: histonet@lists.utsouthwestern.edu Message-ID: <113020042224.12979.41ACF331000E7125000032B32207001641CECECD02019C9D0A9F02@comcast.net> Content-Type: text/plain Netters, Is anyone using an antibody against CD 133 on FFPE tissues? If so, what have you used for a positive control, that could also provide cells to use as a positive control for Flow? I would really appreciate any information anyone would be willing to share. Thanks, Nancy Lemke Hermelin Brain Tumor Center Henry Ford Hospital Detroit ------------------------------ Message: 20 Date: Tue, 30 Nov 2004 17:01:52 -0600 From: "Johnson, Teri" Subject: RE: [Histonet] Paraffin processing explant cultures To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" Liz is a genius. I called Millipore and their customer service guy, Rich, told me I could paraffin process and section these with no problems. Why didn't *I* think of doing this? Sheesh... Teri -----Original Message----- From: Elizabeth Chlipala [mailto:liz@premierlab.com] Sent: Tuesday, November 30, 2004 4:03 PM To: Johnson, Teri; 'Histonet' Subject: RE: [Histonet] Paraffin processing explant cultures Teri I have processed explant cultures with the Millipore MillicellT CM which is a polytetrafluoroethylene based polymer. I process these as any other tissue. Millipore has a great web page in which you can ask questions. I would give them a call about the particular filter that you are using. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Tuesday, November 30, 2004 2:14 PM To: Histonet Subject: [Histonet] Paraffin processing explant cultures Is anybody doing paraffin processing on explant cultures and can give me some tips on increasing my chances for success? The culture plate inserts are Millicell polycarbonate and my question is whether they can survive tissue processing and paraffin sectioning. If not, what are your recommendations for doing these? Thanks in advance! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: Tue, 30 Nov 2004 15:29:32 -0800 From: "Robyn Vazquez" Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? To: LuckG@empirehealth.org, Rcartun@harthosp.org, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Richard, They are a regulated class 6.2, it is a substance known or reasonalbly expected to contain pathogens. If you ship dangerous goods you will need a shipper's declaration which is required. Robyn OHSU >>> "Luck, Greg D." 11/30/2004 1:59:19 PM >>> Richard, Would you further define or point me towards a regulatory definition of "Dangerous Goods". Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 8:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 22 Date: Tue, 30 Nov 2004 19:13:50 -0500 (EST) From: "SMITH,REBEKAH FELICIA" Subject: Re: [Histonet] Ammonia water pH To: "Jackie M O'Connor" , Tim Webster Cc: histonet@lists.utsouthwestern.edu Message-ID: <934672373.1101860030245.JavaMail.osg@osgjas02.cns.ufl.edu> Content-Type: text/plain; format=flowed; charset=us-ascii I agree with Jackie, I use a Coplin jar with distilled water and about 3 drops of 30% ammonium hydroxide, and then rinse in tap water twice. i've never had problems and it produces a rather pretty blue for me. Rebekah Smith On Tue Nov 30 10:35:55 EST 2004, Jackie M O'Connor wrote: > Maybe I'm being extremely simple-minded - but I just fill up a > 200 ml staining jar with tap H20, and add a couple-three drops of > 28%. When it doesn't smell like ammonia anymore, I make new > stuff. How's that grab ya for QA? "Tim Webster" > > Sent by: histonet-bounces@lists.utsouthwestern.edu > 11/30/2004 09:30 AM > > To: > cc: Subject: [Histonet] Ammonia water pH > > > Hi All, > > I'm curious to know what people's procedures are for making and > using ammonia water as a bluing agent in regressive H&E's. > Currently, our methodology is to make the ammonia water up daily > from a 28% stock bottle, and the pH is about 11. My feeling is > that 11 is way to high and that 9-10 is plenty alkaline to blue > sections adequately. Additionally, I'm concerned about strong > alkaline solution increasing the likelihood of sections washing > off or wrinkling. Sheehan & Hrapchak call for a "weak" alkaline > solution in "Theory and Practice of Histotechnology". Also, I can > see no reason not to make a working solution weekly/monthly and > simply empty and refill the Ammonia water station. The pH > doesn't appear to drop off over time. > > Thanks for your time, and have a great week! Tim > > Tim Webster > Histology Specialist > Northwestern Medical Center > 133 Fairfield Street, > St Albans, VT 05478 > (802) 524-1070 (x4349) > 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. >> If you should >> receive > this message in error, please delete it and notify the sender. > 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 > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" ------------------------------ Message: 23 Date: Tue, 30 Nov 2004 16:17:46 -0800 From: Jennifer MacDonald Subject: Re: [Histonet] Paraffin blocks - Dangerous goods? To: "Richard Cartun" Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Advance for Medical Laboratory Professionals had a good article in the November 1, 2004 issue. It is under the Q&A section, titled: Standards for Shipping Infectious Agents. Check www.advanceweb.com to check the archives. If you can't find it, I can fax you a copy. There is also an old article titled "Safe Specimen Packaging Begins with You. It is from the November 12, 2001 issue. Jennifer MacDonald Mt. San Antonio College "Richard Cartun" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2004 08:12 AM To cc Subject [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 24 Date: Tue, 30 Nov 2004 19:26:06 EST From: RCHIOVETTI@aol.com Subject: Re: [Histonet] gliut/isopropyl alcohol To: BMolinari@heart.thi.tmc.edu, histonet@lists.utsouthwestern.edu Message-ID: <1cc.2d22ffb1.2ede699e@aol.com> Content-Type: text/plain; charset="US-ASCII" In a message dated 11/30/2004 12:12:08 PM US Mountain Standard Time, BMolinari@heart.thi.tmc.edu writes: I received samples that have been fixed in a 1%glut / 20% isopropyl alcohol solution for paraffin processing. I have never heard of this fixative, I know alcoholic formalin but never alcoholic glutaraldehyde. Are any of you familiar with this fixative? Thank you. Betsy Molinari HT (ASCP) Hi Betsy, Is the specimen perhaps a heart valve made from pig tissue?? The only time I've heard of alcoholic glutaraldehyde is for pretreating pig valves before they're implanted in humans. There's some kind of relationship between alcohol and glut, and preventing calcification of the valve. Can't remember the details right now, but you have to treat the valve with one of those reagents before using the other. Just a thought... Cheers, Bob Chiovetti ------------------------------ Message: 25 Date: Tue, 30 Nov 2004 17:28:20 -0800 From: "James Watson" Subject: [Histonet] Part time Histology position in San Diego Ca. To: Message-ID: Content-Type: text/plain; charset="US-ASCII" GNF in San Diego has a part time position available. Hours are flexible, 20 hours per week. Please e-mail resume or call me at the number below. The position requires ASCP HT certification. BS, AA degree, H.S. with completion of certified Histology Program with a minimum of 3 years of experience, or equivalent training with a minimum of 6 years of experience. Must be experienced with frozen sectioning, tissue processing and embedding, paraffin sectioning, H&E and special staining, and immunohistochemistry. Animal tissue trimming and identification is preferred. James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org ------------------------------ Message: 26 Date: Wed, 1 Dec 2004 08:18:41 -0000 From: Kemlo Rogerson Subject: RE: [Histonet] Re: Ammonia water pH and other bluing solutions[Sc anned] To: 'Jackie M O'Connor' , "Johnson, Teri" Cc: Histonet , histonet-bounces@lists.utsouthwestern.edu Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B54C@bhrv-nt-11.bhrv.nwest.nhs.uk> Content-Type: text/plain I think you are wrong, so I'll correct you, as you requested. I thought the 'blue' state of haematin was, like litmus, the effect an alkaline condition has on the siderphilic dye. Red, like litmus, denoted acidic conditions. Damned if I know what colour (with the 'u') haematin goes when neutral (bluey/ red?) Running tap water takes longer, I concede, but good things are worth waiting for and it's easier to make up; we rush too much I find. TIP: Never use London water, been through too many kidneys! -----Original Message----- From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: 30 November 2004 19:09 To: Johnson, Teri Cc: Histonet; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions[Scanned] Anyone remember using saturated lithium carbonate? I could probably use a little lithium right now . . . . . Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just bringing the slides BACK to a neutral pH after treating them with acid which makes them purple-ish? I like the ammonia because it 'seems' to make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to make them "blah".(Perception is in the eye of the beholder) Running tap water will make the same miracle happen, but it takes longer. "Johnson, Teri" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2004 12:51 PM To: "Histonet" cc: Subject: [Histonet] Re: Ammonia water pH and other bluing solutions Way back in the day, Brigati was using PBS on his immunostainer to blue after the hematoxylin counterstain. I have since used PBS to blue my slides instead of ammonium hydroxide/water. It's cheap, and very easy on the sections, and I have plenty of it on hand. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 27 Date: Wed, 1 Dec 2004 08:25:23 -0000 From: Kemlo Rogerson Subject: RE: [Histonet] gliut/isopropyl alcohol[Scanned] To: histonet@lists.utsouthwestern.edu Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B54D@bhrv-nt-11.bhrv.nwest.nhs.uk> Content-Type: text/plain Expect the tissue is rather brittle, used to be used for electromicroscopy I think. Wonder if you could secondary fix? Can't aldehyde/ alcohol fixation be 'washed out' then the tissue refixed in something more conventional? -----Original Message----- From: Molinari, Betsy [mailto:BMolinari@heart.thi.tmc.edu] Sent: 30 November 2004 19:08 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] gliut/isopropyl alcohol[Scanned] Hi histonetters, I received samples that have been fixed in a 1%glut / 20% isopropyl alcohol solution for paraffin processing. I have never heard of this fixative, I know alcoholic formalin but never alcoholic glutaraldehyde. Are any of you familiar with this fixative? Thank you. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 28 Date: Wed, 1 Dec 2004 06:18:48 -0600 From: "Molinari, Betsy" Subject: RE: [Histonet] gliut/isopropyl alcohol To: , Message-ID: Content-Type: text/plain; charset="us-ascii" Hi Bob, Thanks for your input. I may ask a couple of the surgery staff here if they are familiar with that technique. A point of interest, these are valves. I loaded them last night starting in 70% ETOH, I am curious to see how they turned out and how much effect it will have on the B&B and PTAH I will be running on them Thanks again, Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology Houston, TX 77030 832-355-6524 -----Original Message----- From: RCHIOVETTI@aol.com [mailto:RCHIOVETTI@aol.com] Sent: Tuesday, November 30, 2004 6:26 PM To: Molinari, Betsy; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] gliut/isopropyl alcohol In a message dated 11/30/2004 12:12:08 PM US Mountain Standard Time, BMolinari@heart.thi.tmc.edu writes: I received samples that have been fixed in a 1%glut / 20% isopropyl alcohol solution for paraffin processing. I have never heard of this fixative, I know alcoholic formalin but never alcoholic glutaraldehyde. Are any of you familiar with this fixative? Thank you. Betsy Molinari HT (ASCP) Hi Betsy, Is the specimen perhaps a heart valve made from pig tissue?? The only time I've heard of alcoholic glutaraldehyde is for pretreating pig valves before they're implanted in humans. There's some kind of relationship between alcohol and glut, and preventing calcification of the valve. Can't remember the details right now, but you have to treat the valve with one of those reagents before using the other. Just a thought... Cheers, Bob Chiovetti ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 13, Issue 1 *************************************** From Barry.R.Rittman <@t> uth.tmc.edu Wed Dec 1 09:18:42 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Re: Ammonia water pH and other bluing solutions Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0010DF214@UTHEVS3.mail.uthouston.edu> When I was training (I admit this was soon after Noah landed the ark), we used tap water to blue sections. This was in London and the water was slightly alkaline at that time. Blueing solutions should be used with great care. Most blueing solutions give essentially the same result. There has been a lot of discussion re using lithium carbonate and ammonia solutions, however very little discussion regarding the dilutions at which these should be used. The smallest amount of a "blueing" agent that will result in blueing of the hematoxylin should be used. It should then be carefully washed to remove all traces. If the solution is too strong it can remove some of the hematoxylin, tend to lift the section off the slide and if not washed out carefully will interfere with the binding of eosin. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen Roberts Sent: Wednesday, December 01, 2004 8:35 AM To: Jackie M O'Connor; 'histonet@lists.utsouthwestern.edu' Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions Heck, we still use that. All our slides look lovely. The mellow one- Kathleen Roberts Rutgers University Jackie M O'Connor wrote: >Anyone remember using saturated lithium carbonate? I could probably use a >little lithium right now . . . . . > >Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just >bringing the slides BACK to a neutral pH after treating them with acid >which makes them purple-ish? I like the ammonia because it 'seems' to >make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to >make them "blah".(Perception is in the eye of the beholder) Running tap >water will make the same miracle happen, but it takes longer. > > > > > > >"Johnson, Teri" >Sent by: histonet-bounces@lists.utsouthwestern.edu >11/30/2004 12:51 PM > > > To: "Histonet" > cc: > Subject: [Histonet] Re: Ammonia water pH and other bluing solutions > > >Way back in the day, Brigati was using PBS on his immunostainer to blue >after the hematoxylin counterstain. I have since used PBS to blue my >slides instead of ammonium hydroxide/water. It's cheap, and very easy >on the sections, and I have plenty of it on hand. > > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, Missouri 64110 >tjj@stowers-institute.org > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Dec 1 09:29:56 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Type of hematoxylin stain with your bluing methods? Tell all, please. In-Reply-To: <003a01c4d727$e077a3d0$eeeea8c0@server> References: <003a01c4d727$e077a3d0$eeeea8c0@server> Message-ID: <6.0.0.22.1.20041201082749.01b372e0@gemini.msu.montana.edu> Dear All, It would be appreciated that you say which hematoxylin you are using when you give your bluing method. For Gudrun, are you using Mayers? At 02:59 PM 11/30/2004, you wrote: >Using lithiumcarbonat is the way we do it with staining frozen sections. >In the HE-stainer we have running tapwater. And the sections are nice. >After haemalaun in special stains we let the slides in tapwater for 3-5 min. > >Gudrun Lang Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From froyer <@t> bitstream.net Wed Dec 1 09:40:33 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods?] Message-ID: <41ADE5F1.9050009@bitstream.net> Cremated human remains are not considered a biohazard, providing that proper documentation from a certified & licensed crematory are presented. The U.S. Postal Service accepts them for shipping as does UPS & FedEx. USPS requires that they be shipped "Certified Mail" but that has to do with insuring that the remains of your loved one gets to their destination safely, not that it would be a Biohazard. On the other hand, paraffin embedded tissue (human or animal) blocks are. Certain viruses and infectious prions can survive fixation and processing (Biohazard) and the paraffin block itself is a flammable material (HazMat). There are ways that they can be shipped if they are properly packaged and labeled by a trained packaging people. ~ Ford Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN 55301 Jackie M O'Connor wrote: >Unfortunately, they are considered a biohazard -(human tissue), although >my sister unknowingly sent our dead aunt's remains (ashes) via UPS once. I >came home from work and she was right there in a little box, sitting on my >front porch. Anyway, I digress - I remember this being discussed before >as to why they (blocks) are still biohazardous since they're fixed and >processed. I'm facing storage problems right now because we don't have >anywhere 'appropriate' to keep them. Years (and years) ago - we stored >them in the basements and sub-basements (scary 4 foot ceilings) of >hospitals until they were found by lucky rodents or other vermin. I'm >curious as to how and where people store them now, as well. > >Jackie > > > > >"Richard Cartun" >Sent by: histonet-bounces@lists.utsouthwestern.edu >11/30/2004 10:12 AM > > > To: > cc: > Subject: [Histonet] Paraffin blocks - Dangerous goods? > > >I have just been told that paraffin tissue blocks are considered >"Dangerous Goods" and can only be packaged and shipped by individuals >who have received special training. Would anyone like to comment on >this? Thank you. > >Richard > > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 >(860) 545-1596 >(860) 545-0174 Fax > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From tissuearray <@t> hotmail.com Wed Dec 1 10:08:45 2004 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Re: Tissue Microarrayers Message-ID: From GAshton <@t> PICR.man.ac.uk Wed Dec 1 10:14:50 2004 From: GAshton <@t> PICR.man.ac.uk (Garry Ashton) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] (no subject) Message-ID: Message: 2 Date: Tue, 30 Nov 2004 13:21:51 -0500 From: Susan Q Wells Subject: [Histonet] Frozen cell pellet To: Histonet@lists.utsouthwestern.edu Message-ID: <41ACBA3F.9030602@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Does anyone have advise on how to freeze cells that have been spun down in a micro centrifuge tube? In the past I've snap frozen the tube, loosened the cells by flicking the tube and then embedded the pellet in OCT. The cells look OK, but the cells in the middle don't look as good as the ones on the perimeter.Any advise would be appreciated. Thanks, Sue Wells Hi Sue, When I freeze cell pellets down that have been rinsed in PBS I remove them from the ependorf using a wide pasteur and place them in OCT, in effect to remove the PBS. I place them in a second OCT "bath" for a futher rinse and then snap freeze in more OCT on a chuck. I think the important thing is firstly they are not to densley packed, and secondly the PBS is replaced by OCT. Hope this helps. Garry PICR UK -------------------------------------------------------- This email is confidential and intended solely for the use of the person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute for Cancer Research or the Christie Hospital NHS Trust. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From Mackinnon <@t> holburn.com Wed Dec 1 10:20:02 2004 From: Mackinnon <@t> holburn.com (John Mackinnon) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods? Message-ID: I would say that that regulation would be open to interpretation. My interpretation would be that most tissues following fixation do not contain viable pathogens and in most cases it would be reasonable to expect that it does not contain pathogens and is therefore not a biohazard. I know that there are some infectious agents that survive fixation and processing but in most cases we are aware of these because they have been diagnosed (most of the time). These few cases fit the definition and should be handled accordingly. This is just one mans opinion. P.S. I wonder if all of the meat packing plants include a shipper's declaration (E.coli, BSE) with their raw unfixed carcases they ship to the grocery stores and butcher shops. Now there is something to think about. John MacKinnon MLT, ART Laboratory Director Holburn Biomedical Corporation 905-623-1484 mackinnon@holburn.com This transmission may contain information that is privileged, confidential and/or exempt from disclosure under applicable law. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or use of the information contained herein (including any reliance thereon) is STRICTLY PROHIBITED. If you received this transmission in error, please immediately contact the sender and destroy the material in its entirety, whether in electronic or hard copy format. Thank you. -----Original Message----- From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] Sent: November 30, 2004 6:30 PM To: LuckG@empirehealth.org; Rcartun@harthosp.org; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Richard, They are a regulated class 6.2, it is a substance known or reasonalbly expected to contain pathogens. If you ship dangerous goods you will need a shipper's declaration which is required. Robyn OHSU >>> "Luck, Greg D." 11/30/2004 1:59:19 PM >>> Richard, Would you further define or point me towards a regulatory definition of "Dangerous Goods". Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 8:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 1 10:30:30 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Type of hematoxylin stain with your bluing methods ? Tell all, please. [Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B55C@bhrv-nt-11.bhrv.nwest.nhs.uk> Harris's and Gill's. -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 01 December 2004 15:30 To: Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Type of hematoxylin stain with your bluing methods? Tell all, please. [Scanned] Dear All, It would be appreciated that you say which hematoxylin you are using when you give your bluing method. For Gudrun, are you using Mayers? At 02:59 PM 11/30/2004, you wrote: >Using lithiumcarbonat is the way we do it with staining frozen sections. >In the HE-stainer we have running tapwater. And the sections are nice. >After haemalaun in special stains we let the slides in tapwater for 3-5 min. > >Gudrun Lang Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bruno.lectard <@t> cea.fr Wed Dec 1 10:34:45 2004 From: bruno.lectard <@t> cea.fr (LECTARD Bruno 172070) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Immunostaining Message-ID: <014DAB1C8A8DD611B60F0008C75DB91C0522D268@oldtrafford.far.cea.fr> I have tried to get the following antibodies working, but have not had any success. Rat Retinoblastoma Protein (RB) -from Santa Cruz Biotechnology, Inc. Cat # SC-7905 and SC -102, Caveoline 1 SC-894 and Caveolin 2 SC-1858. I have followed suggestions on the spec sheets, tried these with and without antigen retrieval. I would like to hear from anyone having experience with these antibodies. If the antibody did work, please send a protocol. Also, I am interested in knowing what other companies supply these antibodies and if anyone has implemented these techniques. Thanks Bruno Lectard * CEA - Fontenay aux roses DSV - DRR - SRCA - LCE BP 6 92265 Fontenay aux roses CEDEX From Bauer.Karen <@t> mayo.edu Wed Dec 1 11:41:48 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods? Message-ID: Our safety person in the lab has just "trained" us all on the proper packaging of specimens in the lab based on the new regulations that are out. Regulations come from the International Civil Aviation Organization (ICAO), the International Air Transport Association (IATA), the USPS, and the Department of Transportation (DOT). Risk groups are ranged from 1 to 4 with 1 being "No to Low Risk" and 4 being "High Risk". Low risk specimens are things like diagnostic serum, whole blood, urine, or tissue samples not known to contain pathogens. Packaging for these specimens need to have a primary container (a bio-hazard bag would be fine for tissue blocks, or maybe the paraffin block itself is the primary container since the tissue is embedded inside it. I'll have to check on that...), then a secondary container (which could be another bio-hazard bag), and then an outer package that is sturdy. This could be a cardboard box or one of those insulated manila envelopes used for fragile items. We do not need to really worry about the outer package, since the couriers that pick up our specimens from the lab have coolers or bio-hazard containers that they put the specimens into. This is considered the "outer package". Our safety person then had everyone who went through the presentation take a short quiz (for competency) and gave us a "Certificate" of being trained. This training will need to be done every 2 years. Everyone in the lab is being trained, since at some point we will all need to know how to ship something out whether it will be Diagnostic or Infectious Specimens (Risk 1 to Risk 4). Karen Bauer HT(ASCP Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 10:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From BennettW <@t> pac.dfo-mpo.gc.ca Wed Dec 1 11:47:05 2004 From: BennettW <@t> pac.dfo-mpo.gc.ca (BennettW@pac.dfo-mpo.gc.ca) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Enzyme stains Message-ID: <7CBBD627E4E688499349A5D11D078316020DBAEE@msgpacpbs.rhq.pac.dfo-mpo.gc.ca> Hello Histonetters, Does anyone know if there is anything that will stain an enzyme called cathepsin. This is an enzyme that is produced by a myxobilatus sp. parasite (Kudoa thyristes) found in fish. The parasite produces this protealytic enzyme. My preliminary searches haven't revealed too much more than that. Any help would be great. Cheers Bill Bennett Histologist Fisheries and Oceans Canada From gu.lang <@t> gmx.at Wed Dec 1 12:35:38 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Re: Type of hematoxylin stain with your bluing methods? Tell all, please. References: <003a01c4d727$e077a3d0$eeeea8c0@server> <6.0.0.22.1.20041201082749.01b372e0@gemini.msu.montana.edu> Message-ID: <003601c4d7d4$8e3bd160$eeeea8c0@server> Harris - frozen sections Mayers - HE stainer Gudrun ----- Original Message ----- From: "Gayle Callis" To: "Gudrun Lang" ; Sent: Wednesday, December 01, 2004 4:29 PM Subject: Type of hematoxylin stain with your bluing methods? Tell all, please. > Dear All, > > It would be appreciated that you say which hematoxylin you are using when > you give your bluing method. For Gudrun, are you using Mayers? > > At 02:59 PM 11/30/2004, you wrote: > >Using lithiumcarbonat is the way we do it with staining frozen sections. > >In the HE-stainer we have running tapwater. And the sections are nice. > >After haemalaun in special stains we let the slides in tapwater for 3-5 min. > > > >Gudrun Lang > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > From kgrobert <@t> rci.rutgers.edu Wed Dec 1 12:55:27 2004 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Type of hematoxylin stain with your bluing methods? Tell all, please. In-Reply-To: <6.0.0.22.1.20041201082749.01b372e0@gemini.msu.montana.edu> References: <003a01c4d727$e077a3d0$eeeea8c0@server> <6.0.0.22.1.20041201082749.01b372e0@gemini.msu.montana.edu> Message-ID: <41AE139F.20203@rci.rutgers.edu> We use Gill's formulation #3 from Fisher Scientific. Kathleen Gayle Callis wrote: > Dear All, > > It would be appreciated that you say which hematoxylin you are using > when you give your bluing method. For Gudrun, are you using Mayers? > > At 02:59 PM 11/30/2004, you wrote: > >> Using lithiumcarbonat is the way we do it with staining frozen sections. >> In the HE-stainer we have running tapwater. And the sections are nice. >> After haemalaun in special stains we let the slides in tapwater for >> 3-5 min. >> >> Gudrun Lang > > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Bauer.Karen <@t> mayo.edu Wed Dec 1 12:51:14 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods? Message-ID: Richard, The primary container does NOT NEED to be a bio-hazard bag. Sorry, I just used that as a suggestion. I did ask our safety person about the paraffin block being the "primary container", but she thought that a plain (small) bag could be used as the primary container and then a bio-hazard bag as the secondary container, since the secondary container is required to have "bio-hazard" on the outside. I realize that "we" handle blocks without gloves all the time, so I'm not sure what to tell you about that. I guess you'll have to check with your infection control person (I'd hate to open a "can-o-worms" here). The regulations are more for the transporting personnel so they know how to handle certain "packages". (That's just what I think...) Hope this helps... Karen -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Wednesday, December 01, 2004 11:50 AM To: Bauer, Karen L. Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Thank you for the detailed information. One question - If you put the paraffin block into a "Biohazard" bag, doesn't that mean that the person who opens the bag needs to be wearing gloves? We (histotechs, lab aides, secretaries, pathologists, etc.) handle paraffin tissue blocks all the time without wearing gloves. RWC 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 >>> "Bauer, Karen" 12/01/04 12:41PM >>> Our safety person in the lab has just "trained" us all on the proper packaging of specimens in the lab based on the new regulations that are out. Regulations come from the International Civil Aviation Organization (ICAO), the International Air Transport Association (IATA), the USPS, and the Department of Transportation (DOT). Risk groups are ranged from 1 to 4 with 1 being "No to Low Risk" and 4 being "High Risk". Low risk specimens are things like diagnostic serum, whole blood, urine, or tissue samples not known to contain pathogens. Packaging for these specimens need to have a primary container (a bio-hazard bag would be fine for tissue blocks, or maybe the paraffin block itself is the primary container since the tissue is embedded inside it. I'll have to check on that...), then a secondary container (which could be another bio-hazard bag), and then an outer package that is sturdy. This could be a cardboard box or one of those insulated manila envelopes used for fragile items. We do not need to really worry about the outer package, since the couriers that pick up our specimens from the lab have coolers or bio-hazard containers that they put the specimens into. This is considered the "outer package". Our safety person then had everyone who went through the presentation take a short quiz (for competency) and gave us a "Certificate" of being trained. This training will need to be done every 2 years. Everyone in the lab is being trained, since at some point we will all need to know how to ship something out whether it will be Diagnostic or Infectious Specimens (Risk 1 to Risk 4). Karen Bauer HT(ASCP Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 10:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From Stacy_McLaughlin <@t> cooley-dickinson.org Wed Dec 1 13:06:02 2004 From: Stacy_McLaughlin <@t> cooley-dickinson.org (Stacy McLaughlin) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods? Message-ID: <3D502BBF5356D31184650090275B750D0346C892@mail.cooley-dickinson.org> Is anyone checking that this requirement is being fulfilled? (CAP, JCAHO, OSHA, etc...) I want to make sure the right thing is being done, but I also would like to know if anyone is watching for that specifically. Thanks, Stacy McLaughlin HT (ASCP) (and Lab Safety Lady) Cooley Dickinson Hospital Northampton,MA -----Original Message----- From: Bauer, Karen [mailto:Bauer.Karen@mayo.edu] Sent: Wednesday, December 01, 2004 12:42 PM To: 'Richard Cartun'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Our safety person in the lab has just "trained" us all on the proper packaging of specimens in the lab based on the new regulations that are out. Regulations come from the International Civil Aviation Organization (ICAO), the International Air Transport Association (IATA), the USPS, and the Department of Transportation (DOT). Risk groups are ranged from 1 to 4 with 1 being "No to Low Risk" and 4 being "High Risk". Low risk specimens are things like diagnostic serum, whole blood, urine, or tissue samples not known to contain pathogens. Packaging for these specimens need to have a primary container (a bio-hazard bag would be fine for tissue blocks, or maybe the paraffin block itself is the primary container since the tissue is embedded inside it. I'll have to check on that...), then a secondary container (which could be another bio-hazard bag), and then an outer package that is sturdy. This could be a cardboard box or one of those insulated manila envelopes used for fragile items. We do not need to really worry about the outer package, since the couriers that pick up our specimens from the lab have coolers or bio-hazard containers that they put the specimens into. This is considered the "outer package". Our safety person then had everyone who went through the presentation take a short quiz (for competency) and gave us a "Certificate" of being trained. This training will need to be done every 2 years. Everyone in the lab is being trained, since at some point we will all need to know how to ship something out whether it will be Diagnostic or Infectious Specimens (Risk 1 to Risk 4). Karen Bauer HT(ASCP Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 10:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet THIS E-MAIL MESSAGE IS INTENDED ONLY FOR THE USE OF THE INDIVIDUAL OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If the reader of this e-mail 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 e-mail message is strictly prohibited. If you have received this e-mail message in error, please immediately notify Cooley Dickinson Healthcare at 413-582-2000 and delete or shred the original message and all copies thereof. Thank you. From vazquezr <@t> ohsu.edu Wed Dec 1 13:15:09 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods? Message-ID: John, Now that I think about it, I get blocks in the mail occasionally. Maybe it is a regulation that is not known to a doctors office or it is a turn blind eye regulation, because it's not like getting a slab of meat in the mail ;0) Robyn OHSU >>> "John Mackinnon" 12/1/2004 8:20:02 AM >>> I would say that that regulation would be open to interpretation. My interpretation would be that most tissues following fixation do not contain viable pathogens and in most cases it would be reasonable to expect that it does not contain pathogens and is therefore not a biohazard. I know that there are some infectious agents that survive fixation and processing but in most cases we are aware of these because they have been diagnosed (most of the time). These few cases fit the definition and should be handled accordingly. This is just one mans opinion. P.S. I wonder if all of the meat packing plants include a shipper's declaration (E.coli, BSE) with their raw unfixed carcases they ship to the grocery stores and butcher shops. Now there is something to think about. John MacKinnon MLT, ART Laboratory Director Holburn Biomedical Corporation 905-623-1484 mackinnon@holburn.com This transmission may contain information that is privileged, confidential and/or exempt from disclosure under applicable law. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or use of the information contained herein (including any reliance thereon) is STRICTLY PROHIBITED. If you received this transmission in error, please immediately contact the sender and destroy the material in its entirety, whether in electronic or hard copy format. Thank you. -----Original Message----- From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] Sent: November 30, 2004 6:30 PM To: LuckG@empirehealth.org; Rcartun@harthosp.org; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Richard, They are a regulated class 6.2, it is a substance known or reasonalbly expected to contain pathogens. If you ship dangerous goods you will need a shipper's declaration which is required. Robyn OHSU >>> "Luck, Greg D." 11/30/2004 1:59:19 PM >>> Richard, Would you further define or point me towards a regulatory definition of "Dangerous Goods". Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 8:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Wed Dec 1 13:16:24 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Type of hematoxylin stain with your bluing methods ? Tell all, please. [Scanned] Message-ID: Shandon Instant Hematoxylin >>> Kemlo Rogerson 12/1/2004 8:30:30 AM >>> Harris's and Gill's. -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 01 December 2004 15:30 To: Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Type of hematoxylin stain with your bluing methods? Tell all, please. [Scanned] Dear All, It would be appreciated that you say which hematoxylin you are using when you give your bluing method. For Gudrun, are you using Mayers? At 02:59 PM 11/30/2004, you wrote: >Using lithiumcarbonat is the way we do it with staining frozen sections. >In the HE-stainer we have running tapwater. And the sections are nice. >After haemalaun in special stains we let the slides in tapwater for 3-5 min. > >Gudrun Lang Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Wed Dec 1 14:00:00 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Enzyme stains Message-ID: Novocastra has a few different antibodies for cathepsin. www.novocastra.co.uk 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: BennettW@pac.dfo-mpo.gc.ca [mailto:BennettW@pac.dfo-mpo.gc.ca] Sent: Wednesday, December 01, 2004 11:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Enzyme stains Hello Histonetters, Does anyone know if there is anything that will stain an enzyme called cathepsin. This is an enzyme that is produced by a myxobilatus sp. parasite (Kudoa thyristes) found in fish. The parasite produces this protealytic enzyme. My preliminary searches haven't revealed too much more than that. Any help would be great. Cheers Bill Bennett Histologist Fisheries and Oceans Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NSEARCY <@t> swmail.sw.org Wed Dec 1 14:21:16 2004 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Milestone Microwave Processor Message-ID: After 18 months in service, we are experiencing paraffin being drawn into the vacuum tube - anyone else having this problem? Can anyone provide help ? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:4-2438 ORG:;Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD From Bauer.Karen <@t> mayo.edu Wed Dec 1 14:27:40 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods? Message-ID: Stacy, If you go to the CAP TODAY, Oct. 2004 issue on page 101, there is a Q & A about shipping of materials. It is also on the CAP General Checklist # GEN.40522. CAP will be changing their training requirements to every two years instead of the "annual training" that they have on the checklist now. Hope this helps, Karen -----Original Message----- From: Stacy McLaughlin [mailto:Stacy_McLaughlin@cooley-dickinson.org] Sent: Wednesday, December 01, 2004 1:06 PM To: 'Bauer, Karen'; 'Richard Cartun'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Is anyone checking that this requirement is being fulfilled? (CAP, JCAHO, OSHA, etc...) I want to make sure the right thing is being done, but I also would like to know if anyone is watching for that specifically. Thanks, Stacy McLaughlin HT (ASCP) (and Lab Safety Lady) Cooley Dickinson Hospital Northampton,MA -----Original Message----- From: Bauer, Karen [mailto:Bauer.Karen@mayo.edu] Sent: Wednesday, December 01, 2004 12:42 PM To: 'Richard Cartun'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Our safety person in the lab has just "trained" us all on the proper packaging of specimens in the lab based on the new regulations that are out. Regulations come from the International Civil Aviation Organization (ICAO), the International Air Transport Association (IATA), the USPS, and the Department of Transportation (DOT). Risk groups are ranged from 1 to 4 with 1 being "No to Low Risk" and 4 being "High Risk". Low risk specimens are things like diagnostic serum, whole blood, urine, or tissue samples not known to contain pathogens. Packaging for these specimens need to have a primary container (a bio-hazard bag would be fine for tissue blocks, or maybe the paraffin block itself is the primary container since the tissue is embedded inside it. I'll have to check on that...), then a secondary container (which could be another bio-hazard bag), and then an outer package that is sturdy. This could be a cardboard box or one of those insulated manila envelopes used for fragile items. We do not need to really worry about the outer package, since the couriers that pick up our specimens from the lab have coolers or bio-hazard containers that they put the specimens into. This is considered the "outer package". Our safety person then had everyone who went through the presentation take a short quiz (for competency) and gave us a "Certificate" of being trained. This training will need to be done every 2 years. Everyone in the lab is being trained, since at some point we will all need to know how to ship something out whether it will be Diagnostic or Infectious Specimens (Risk 1 to Risk 4). Karen Bauer HT(ASCP Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 10:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet THIS E-MAIL MESSAGE IS INTENDED ONLY FOR THE USE OF THE INDIVIDUAL OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If the reader of this e-mail 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 e-mail message is strictly prohibited. If you have received this e-mail message in error, please immediately notify Cooley Dickinson Healthcare at 413-582-2000 and delete or shred the original message and all copies thereof. Thank you. ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From REEVEML <@t> shands.ufl.edu Wed Dec 1 15:39:01 2004 From: REEVEML <@t> shands.ufl.edu (Mary Reeves) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Milestone Microwave Processor Message-ID: Is the paraffin level too high? If we have more cassettes than normal (it pushes the level higher) we always pour off some of the paraffin prior to processing. Mary Reeves Lab Client Operations Coordinator Shands Medical Laboratories >>> "Nita Searcy" 12/1/2004 3:21:16 PM >>> After 18 months in service, we are experiencing paraffin being drawn into the vacuum tube - anyone else having this problem? Can anyone provide help ? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 From Laurie.Pereira <@t> sdcounty.ca.gov Wed Dec 1 17:17:38 2004 From: Laurie.Pereira <@t> sdcounty.ca.gov (Pereira, Laurie ) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] static electricity Message-ID: Hey all, I'm new at Histology and am having a problem with static electricity. During the winter, everything tends to stick to me. I've heard that a humidifier works pretty well but our laboratory is pretty large. Will a humidifier work in such a large space? Does anyone have any hints on how to deal with the wonders of static electricity? Any information would be extremely helpful! Thanks Frustrated From JWEEMS <@t> sjha.org Wed Dec 1 17:35:46 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] static electricity Message-ID: <83AACDB0810528418AA106F9AE9B7F7E5075EB@sjhaexc02.sjha.org> Use a dryer sheet to wipe the microtome and yourself and the area around the microtome. This often helps. Good luck! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Pereira, Laurie Sent: Wednesday, December 01, 2004 6:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] static electricity Hey all, I'm new at Histology and am having a problem with static electricity. During the winter, everything tends to stick to me. I've heard that a humidifier works pretty well but our laboratory is pretty large. Will a humidifier work in such a large space? Does anyone have any hints on how to deal with the wonders of static electricity? Any information would be extremely helpful! Thanks Frustrated _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Laurie.Pereira <@t> sdcounty.ca.gov Wed Dec 1 18:04:55 2004 From: Laurie.Pereira <@t> sdcounty.ca.gov (Pereira, Laurie ) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] re:static electricity Message-ID: Thanks to all So much frustration for such a simple solution. I never would have thought of it! I'm going out to get static sheets tonight. Thanks again! From Shrttmprd <@t> aol.com Wed Dec 1 18:06:57 2004 From: Shrttmprd <@t> aol.com (Shrttmprd@aol.com) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] A good illiustrated book Message-ID: <193.340485dd.2edfb6a1@aol.com> Could anyone recommend a good illustrated book for slides with special stains and so forth? I wanted a good reference for myself on the types of stains on various tissues. Thank you in advance for your support. Oh! Thank you to everyone that gave me sound advice on preparing for the HT practical! I did pass the practical but am still waiting for the results on the slide submissions. So, I am not out of the woods yet and and my fingers are still crossed! I will keep you posted once I hear something. From rockbeki <@t> ufl.edu Wed Dec 1 19:18:53 2004 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Type of hematoxylin stain with your bluing methods ? Tell all, please. [Scanned] Message-ID: <1423675899.1101950333006.JavaMail.osg@osgjas02.cns.ufl.edu> I use Gill's. (I use it as a counterstain to AEC, so I needed to use something without alcohol) -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" From bettina.hutz <@t> orionpharma.com Wed Dec 1 23:26:22 2004 From: bettina.hutz <@t> orionpharma.com (bettina.hutz@orionpharma.com) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] static electricity Message-ID: <243E1A79AF5D89438C70513DBADAA4F0019E3FF4@sfies-exchange1.orionnet.org> Hello, I have the same problem during winter time - and in summer time it is the opposite: it is "common accepted" that in July/August there is hardly any staining possible, because the huminidy goes immediately to the alcohol and then the xylene is milky and mounting not proper. When I have been working in Germany for 10 years I never has this effect, but since I work in Finland I am fighting with it every summer. So, if anyone has a solution to the humidity problem, please share it with me:) Tina -----Original Message----- From: Pereira, Laurie [mailto:Laurie.Pereira@sdcounty.ca.gov] Sent: Thursday, December 02, 2004 1:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] static electricity Hey all, I'm new at Histology and am having a problem with static electricity. During the winter, everything tends to stick to me. I've heard that a humidifier works pretty well but our laboratory is pretty large. Will a humidifier work in such a large space? Does anyone have any hints on how to deal with the wonders of static electricity? Any information would be extremely helpful! Thanks Frustrated _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed Dec 1 23:28:37 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Enzyme stains References: <7CBBD627E4E688499349A5D11D078316020DBAEE@msgpacpbs.rhq.pac.dfo-mpo.gc.ca> Message-ID: <41AEA805.B0A76F8D@uwo.ca> Cathepsins are proteolytic enzymes. They occur in all organisms. A big biochemistry textbook will tell you about these enzymes. Some cathepsins can be histochemically localized by activity-based histochemical methods. The best reference is Volume 3 of the last edition of Pearse's Histochemistry: ISBN 0443029962 You will need to study Chapters 29 and 31. You may need to follow up some references from the late Pearse's Vol 3, but that is now extremely easy, with PubMed, Google and Web of Science. John Kiernan London, Canada. ---------------- BennettW@pac.dfo-mpo.gc.ca wrote: > > Hello Histonetters, > > Does anyone know if there is anything that will stain an enzyme called > cathepsin. This is an enzyme that is produced by a myxobilatus sp. parasite > (Kudoa thyristes) found in fish. The parasite produces this protealytic > enzyme. My preliminary searches haven't revealed too much more than that. > > Any help would be great. > > Cheers > Bill Bennett > Histologist > Fisheries and Oceans Canada > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Thu Dec 2 02:29:37 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] static electricity[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B55E@bhrv-nt-11.bhrv.nwest.nhs.uk> Air conditioning? A dehumidifier? -----Original Message----- From: bettina.hutz@orionpharma.com [mailto:bettina.hutz@orionpharma.com] Sent: 02 December 2004 05:26 To: Laurie.Pereira@sdcounty.ca.gov; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] static electricity[Scanned] Hello, I have the same problem during winter time - and in summer time it is the opposite: it is "common accepted" that in July/August there is hardly any staining possible, because the huminidy goes immediately to the alcohol and then the xylene is milky and mounting not proper. When I have been working in Germany for 10 years I never has this effect, but since I work in Finland I am fighting with it every summer. So, if anyone has a solution to the humidity problem, please share it with me:) Tina -----Original Message----- From: Pereira, Laurie [mailto:Laurie.Pereira@sdcounty.ca.gov] Sent: Thursday, December 02, 2004 1:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] static electricity Hey all, I'm new at Histology and am having a problem with static electricity. During the winter, everything tends to stick to me. I've heard that a humidifier works pretty well but our laboratory is pretty large. Will a humidifier work in such a large space? Does anyone have any hints on how to deal with the wonders of static electricity? Any information would be extremely helpful! Thanks Frustrated _______________________________________________ Histonet mailing list Histonet@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 NSEARCY <@t> swmail.sw.org Thu Dec 2 07:50:18 2004 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] New JCAHO PI Standard Message-ID: There is a new standard that states: "An analysis is performed for all major discrepancies between preoperative and postoperative (including pathologic) diagnoses." How are institutions complying ? In my past lives there were "Tissue Committees" that reviewed "normal" tissues but this standard , in my opinion, would not be able to be handled in the pathology lab. We consider ourselves lucky if we get any preoperative diagnosis on our requisitions received from surgery. Perhaps some kind of chart review on all surgeries? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:4-2438 ORG:;Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD From king_of_karaoke <@t> web.de Thu Dec 2 07:54:43 2004 From: king_of_karaoke <@t> web.de (=?iso-8859-1?Q? Tobias=20M=E4tzig ?=) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Autofluorescence problem in olfactory epithelia cryosections Message-ID: <963396644@web.de> Hello, I've got a serious autoflourescence problem when trying to perform ISH on mouse head cryosections. After ISH process I can see a bright red autoflourescence in large parts of my tissue even without any antibodies on it. First I thought it might be due to blocking solution containing Horse serum, but I test blocked my slides and there was almost no autoflourescence at all. So were the untreated slide ! Right now, I don't have a single clue which step of my ISH might cause the red autofluorescence but it would be great if anyone of you had a good advice for me. Please, let me know ! The mouse has been killed, decapitated, parts of the nose were removed and fixed in 4%PFA for 3h. After that the tissue was incubated at 4?C in 30% succrose until the tissue sank to the bottom of the tube and was than shock frozen and embedded in mounting medium. Slices are 10?m thick. Thank you for your help. -Tobias- ________________________________________________________________ Verschicken Sie romantische, coole und witzige Bilder per SMS! Jetzt neu bei WEB.DE FreeMail: http://freemail.web.de/?mc=021193 From bliven.laura <@t> marshfieldclinic.org Thu Dec 2 08:50:37 2004 From: bliven.laura <@t> marshfieldclinic.org (bliven.laura@marshfieldclinic.org) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Tissue Disposal ? Message-ID: <170bf01c4d87e$49349c00$8e05010a@mfldclinframe.org> Because of new regulations, our current incinerator (that we routinely use for disposal of formalin fixed tissue) is to be torned down in the near future. I understand that there are other ways to dispose of tissue and am searching for possibilities and/or systems. Apparently there are crematory-type and pressure cooker-type systems. Any ideas, company info, expense info would be greatly appreciated. Thanks, Laura Bliven Marshfield Laboratory St. Joseph's Hospital Marshfield, WI 54449 bliven.laura@marshfieldclinic.org (715)387-7810 From GDawson <@t> Milw.Dynacare.com Thu Dec 2 08:50:09 2004 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] RE: Autoimmunostainers DAKO vs Ventanna Message-ID: Lori, Great response. I've used both as well and, if you want to take a more hands on approach to IHC and you want to maximize profitability, Dako is the way to go. In my humble opinion, the staining that I got using Dako vs. Ventanna looked better. The histonet archives should be packed with Dako vs. Ventanna discussions BTW. Glen Dawson BS, HT & QIHC (ASCP) IHC Coordinator Milwaukee, WI -----Original Message----- From: White, Lori [mailto:lwhite@lakeridgehealth.on.ca] Sent: Wednesday, December 01, 2004 8:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Autoimmunostainers DAKO vs Ventanna I have used both systems and there are pros and cons to each. It really is dependent on your IHC volume and how you want to use the automation. If you want to be free of working up antibodies from concentrates to maximize dilution and are willing to pay the expense of prediluted reagents, the Ventana system is probably the better way to go. Keep in mind that you are locked into one source for all reagents including buffers and detection. To me, this is a disadvantage. If you have a fairly high volume IHC laboratory and experienced staff, I would recommend the open systems. You have a lot more flexibility for not only primary antibodies but all of your ancillary reagents as well. One of the features I liked about the Dako was that you were able to see what was happening during the staining run, not only on the instrument but on the computer screen as well. I also liked the sensor/alarm for dispensing of reagents. In terms of service, I would say that again both systems have positives and negatives. My two cents...... Lori White Charge Technologist Lakeridge Health Corporation Oshawa, Ontario Canada -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Wednesday, December 01, 2004 7:26 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 13, Issue 1 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. Ammonia water pH (Paula Pierce) 2. Frozen cell pellet (Susan Q Wells) 3. RE: Frozen cell pellet (Favara, Cynthia (NIH/NIAID)) 4. Re: Ammonia water pH and other bluing solutions (Johnson, Teri) 5. gliut/isopropyl alcohol (Molinari, Betsy) 6. Re: Re: Ammonia water pH and other bluing solutions (Jackie M O'Connor) 7. Re: Re: Ammonia water pH and other bluing solutions (Robyn Vazquez) 8. RE: Autoimmunostainers DAKO vs Ventanna[Scanned] (Darren James) 9. Re: Frozen cell pellet (Gayle Callis) 10. Saturated lithium carbonate bluing (Gayle Callis) 11. RE: Autoimmunostainers DAKO vs Ventanna[Scanned] (Darren James) 12. Re: Ammonia water pH and other bluing solutions (Johnson, Teri) 13. Paraffin processing explant cultures (Johnson, Teri) 14. Re: Re: Ammonia water pH and other bluing solutions (Gudrun Lang) 15. RE: Paraffin blocks - Dangerous goods? (Luck, Greg D.) 16. RE: Counterstain for Von Kossa (Fredrik Johansson) 17. Phosphorylated-met antibodies (Tan, MinHan) 18. RE: Paraffin processing explant cultures (Elizabeth Chlipala) 19. CD 133 (nperson211@comcast.net) 20. RE: Paraffin processing explant cultures (Johnson, Teri) 21. RE: Paraffin blocks - Dangerous goods? (Robyn Vazquez) 22. Re: Ammonia water pH (SMITH,REBEKAH FELICIA) 23. Re: Paraffin blocks - Dangerous goods? (Jennifer MacDonald) 24. Re: gliut/isopropyl alcohol (RCHIOVETTI@aol.com) 25. Part time Histology position in San Diego Ca. (James Watson) 26. RE: Re: Ammonia water pH and other bluing solutions[Sc anned] (Kemlo Rogerson) 27. RE: gliut/isopropyl alcohol[Scanned] (Kemlo Rogerson) 28. RE: gliut/isopropyl alcohol (Molinari, Betsy) ---------------------------------------------------------------------- Message: 1 Date: Tue, 30 Nov 2004 10:17:12 -0800 (PST) From: Paula Pierce Subject: [Histonet] Ammonia water pH To: histonet@lists.utsouthwestern.edu Message-ID: <20041130181712.32329.qmail@web50303.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii lol, Jackie O'Conner. I like you. I do the same thing. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com ------------------------------ Message: 2 Date: Tue, 30 Nov 2004 13:21:51 -0500 From: Susan Q Wells Subject: [Histonet] Frozen cell pellet To: Histonet@lists.utsouthwestern.edu Message-ID: <41ACBA3F.9030602@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Does anyone have advise on how to freeze cells that have been spun down in a micro centrifuge tube? In the past I've snap frozen the tube, loosened the cells by flicking the tube and then embedded the pellet in OCT. The cells look OK, but the cells in the middle don't look as good as the ones on the perimeter.Any advise would be appreciated. Thanks, Sue Wells ------------------------------ Message: 3 Date: Tue, 30 Nov 2004 13:38:40 -0500 From: "Favara, Cynthia (NIH/NIAID)" Subject: RE: [Histonet] Frozen cell pellet To: 'Susan Q Wells' , Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain I have done as you have, also spin down add a bit of histogel vortex let cool then flick out and put in OCT. I like this as I seem to get a more even distribution of cells. 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: Susan Q Wells [mailto:susan.wells@bms.com] Sent: Tuesday, November 30, 2004 11:22 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Frozen cell pellet Does anyone have advise on how to freeze cells that have been spun down in a micro centrifuge tube? In the past I've snap frozen the tube, loosened the cells by flicking the tube and then embedded the pellet in OCT. The cells look OK, but the cells in the middle don't look as good as the ones on the perimeter.Any advise would be appreciated. Thanks, Sue Wells _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Tue, 30 Nov 2004 12:51:22 -0600 From: "Johnson, Teri" Subject: [Histonet] Re: Ammonia water pH and other bluing solutions To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" Way back in the day, Brigati was using PBS on his immunostainer to blue after the hematoxylin counterstain. I have since used PBS to blue my slides instead of ammonium hydroxide/water. It's cheap, and very easy on the sections, and I have plenty of it on hand. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org ------------------------------ Message: 5 Date: Tue, 30 Nov 2004 13:07:43 -0600 From: "Molinari, Betsy" Subject: [Histonet] gliut/isopropyl alcohol To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi histonetters, I received samples that have been fixed in a 1%glut / 20% isopropyl alcohol solution for paraffin processing. I have never heard of this fixative, I know alcoholic formalin but never alcoholic glutaraldehyde. Are any of you familiar with this fixative? Thank you. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 ------------------------------ Message: 6 Date: Tue, 30 Nov 2004 13:08:41 -0600 From: "Jackie M O'Connor" Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions To: "Johnson, Teri" Cc: Histonet , histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Anyone remember using saturated lithium carbonate? I could probably use a little lithium right now . . . . . Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just bringing the slides BACK to a neutral pH after treating them with acid which makes them purple-ish? I like the ammonia because it 'seems' to make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to make them "blah".(Perception is in the eye of the beholder) Running tap water will make the same miracle happen, but it takes longer. "Johnson, Teri" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2004 12:51 PM To: "Histonet" cc: Subject: [Histonet] Re: Ammonia water pH and other bluing solutions Way back in the day, Brigati was using PBS on his immunostainer to blue after the hematoxylin counterstain. I have since used PBS to blue my slides instead of ammonium hydroxide/water. It's cheap, and very easy on the sections, and I have plenty of it on hand. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Tue, 30 Nov 2004 11:38:08 -0800 From: "Robyn Vazquez" Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions To: Jackie.O'Connor@abbott.com, histonet-bounces@lists.utsouthwestern.edu, TJJ@Stowers-Institute.org Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Jackie, I use a saturated lithium carbonate for my frozen Mohs tissue. Works and looks great... Robyn OHSU >>> "Jackie M O'Connor" 11/30/2004 11:08:41 AM >>> Anyone remember using saturated lithium carbonate? I could probably use a little lithium right now . . . . . Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just bringing the slides BACK to a neutral pH after treating them with acid which makes them purple-ish? I like the ammonia because it 'seems' to make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to make them "blah".(Perception is in the eye of the beholder) Running tap water will make the same miracle happen, but it takes longer. "Johnson, Teri" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2004 12:51 PM To: "Histonet" cc: Subject: [Histonet] Re: Ammonia water pH and other bluing solutions Way back in the day, Brigati was using PBS on his immunostainer to blue after the hematoxylin counterstain. I have since used PBS to blue my slides instead of ammonium hydroxide/water. It's cheap, and very easy on the sections, and I have plenty of it on hand. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Wed, 1 Dec 2004 08:49:29 +1300 From: "Darren James" Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] To: "'Kemlo Rogerson'" , "Histonet \(E-mail\)" Message-ID: <000001c4d715$b49168d0$c864a8c0@medica.co.nz> Content-Type: text/plain; charset="iso-8859-1" As another option take a look at the Bond Max manufactured by Vision Bio Systems http://www.vision-bio.com/product_bondmax.html It performs on board dewaxing, heat induced epitope retreival, enzyme digestion as well as counterstaining. I have only heard good things about it. I have used the Dako extensively and although good, it has it's limitations. At the time we chose the Dako over the Ventana due to the Ventana being a closed system. If I were in a position to purchase another I would definitely take a long hard look at the Bond Max. Thanks and have a great day Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kemlo Rogerson Sent: Wednesday, 1 December 2004 4:36 a.m. To: histonet@pathology.swmed.edu Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] WE have both, and I'm told the Ventana is the 'better' of the two options. As an ICC virgin and one who will have to choose which platform to go with I am particularly interested in this thread. I would prefer to use the cheapest 'open' system that is a automated as possible, is that mutually exclusive? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Maliniak, Richard [mailto:RMaliniak@SBHCS.com] Sent: 30 November 2004 15:16 To: histonet@pathology.swmed.edu Subject: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] I am looking for info= rmation from people who have switched from Ventanna to DAKO and DAKO to V= entanna. Why did you switch? What did you like or dislike abo= ut one or the other. Rick ________________________________________________________________ Important news about our email communications Saint Barnabas Health Care System has implemented secure messaging servic= es. To learn more about SBHCS Secure Messaging, go to: [1]http://www.zixcorp.c= om/evangelism/sbhcs/ If you need assistance with retrieving a secure email, please email sbhcs= accounts@sbhcs.com or visit [2]http://www.zixcorp.com/evangelism/sbhcs/par= tners/receiving.php References 1. 3D"http://www.zixcorp.com/evangelism/sbhcs/" 2. 3D"http://www.zixcorp.com/evangelism/_______________________________________ ________ Histonet mailing list Histonet@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: 9 Date: Tue, 30 Nov 2004 12:45:41 -0700 From: Gayle Callis Subject: Re: [Histonet] Frozen cell pellet To: Susan Q Wells , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041130123759.01b57008@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed How many cells you spin down may be critical. Here is a protocol that works for us. 1. 1 to 3 x 10 (to the 7th) cells/ml. 2. Spin down and rinse three time with PBS. 3. Add OCT (approx 1 ml or less) to last pellet, mix. 4. Immerse tube directly into liquid N2. 5. Pop block out with a sharp rap inside cryostat. Mount block and section. The key is to make sure you don't have too many cells and do a good mix with OCT in order to disperse any water from buffer and spread out cells. We have sectioned thinner, 4 um rather than 5 to insure uncrowded staining. We build block up with more OCT around end of block so the block face is not so tiny and you can manipulate the tiny section. Chris van der Loos is a master at the supertiny section!! A wider conical end microcentrifuge tube (holds 2 ml or more overall) spreads things out a bit more for a broader faced block but a 1.5 ml tube will work. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 10 Date: Tue, 30 Nov 2004 12:58:26 -0700 From: Gayle Callis Subject: [Histonet] Saturated lithium carbonate bluing To: "Jackie M O'Connor" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041130124819.01b66e58@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Yes, have used sat LiCO3, but it had to be decanted carefully into the container or the lithium carbonate crystals poured off polarized!! Bummer with congo red amyloid staining to see annoying crystals everywhere. And then the saturated stuff grew some ugly black floating grow-ty stuff, a touch of black slime. Went back to Scotts tap water substitute or Richard Allen bluing solution. An excellent article on the theory of hematoxylin staining is found in J of Histotechnology, Susan Meloan and H. Puchtler volume 10, 1987. The actual staining by hematoxylin, nomenclature, and what the dye molecule consists of, along with what acid and alkaline solutions does is explained in great detail. After reading this publication, how I do or think about hematoxylin staining (either progressive or regressive methods) was radically changed. It is very chemical, but very informative - be prepared for some interesting chemistry. I strongly recommend this publication to all doing H&E staining. At 12:08 PM 11/30/2004, you wrote: >Anyone remember using saturated lithium carbonate? I could probably use a >little lithium right now . . . . . > >Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just >bringing the slides BACK to a neutral pH after treating them with acid >which makes them purple-ish? I like the ammonia because it 'seems' to >make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to >make them "blah".(Perception is in the eye of the beholder) Running tap >water will make the same miracle happen, but it takes longer. > > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 11 Date: Wed, 1 Dec 2004 09:28:05 +1300 From: "Darren James" Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] To: "'Bonnie Whitaker'" , "Histonet \(E-mail\)" Message-ID: <000801c4d71b$198aeb80$c864a8c0@medica.co.nz> Content-Type: text/plain; charset="us-ascii" Hi Bonnie, I don't know about the US and the UK but here in NZ and also Australia they are being placed at various sites. I would assume that if there were any pending legal action the hold on selling would be global. Thanks Darren -----Original Message----- From: Bonnie Whitaker [mailto:bwhitaker@brownpathology.com] Sent: Wednesday, 1 December 2004 9:01 a.m. To: darrenj@medica.co.nz Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] I heard that they were held up in litigation, and would be unable to sell the instruments for at least a year..... Also, it's a closed system (at least to some degree.) Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Darren James Sent: Tuesday, November 30, 2004 1:49 PM To: 'Kemlo Rogerson'; Histonet (E-mail) Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] As another option take a look at the Bond Max manufactured by Vision Bio Systems http://www.vision-bio.com/product_bondmax.html It performs on board dewaxing, heat induced epitope retreival, enzyme digestion as well as counterstaining. I have only heard good things about it. I have used the Dako extensively and although good, it has it's limitations. At the time we chose the Dako over the Ventana due to the Ventana being a closed system. If I were in a position to purchase another I would definitely take a long hard look at the Bond Max. Thanks and have a great day Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kemlo Rogerson Sent: Wednesday, 1 December 2004 4:36 a.m. To: histonet@pathology.swmed.edu Subject: RE: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] WE have both, and I'm told the Ventana is the 'better' of the two options. As an ICC virgin and one who will have to choose which platform to go with I am particularly interested in this thread. I would prefer to use the cheapest 'open' system that is a automated as possible, is that mutually exclusive? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Maliniak, Richard [mailto:RMaliniak@SBHCS.com] Sent: 30 November 2004 15:16 To: histonet@pathology.swmed.edu Subject: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] I am looking for info= rmation from people who have switched from Ventanna to DAKO and DAKO to V= entanna. Why did you switch? What did you like or dislike abo= ut one or the other. Rick ________________________________________________________________ Important news about our email communications Saint Barnabas Health Care System has implemented secure messaging servic= es. To learn more about SBHCS Secure Messaging, go to: [1]http://www.zixcorp.c= om/evangelism/sbhcs/ If you need assistance with retrieving a secure email, please email sbhcs= accounts@sbhcs.com or visit [2]http://www.zixcorp.com/evangelism/sbhcs/par= tners/receiving.php References 1. 3D"http://www.zixcorp.com/evangelism/sbhcs/" 2. 3D"http://www.zixcorp.com/evangelism/_______________________________________ ________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Tue, 30 Nov 2004 15:09:05 -0600 From: "Johnson, Teri" Subject: [Histonet] Re: Ammonia water pH and other bluing solutions To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" FYI, the pH of the PBS we use is 7.2 but I'm sure any neutral to slightly alkaline pH would work fine. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org ------------------------------ Message: 13 Date: Tue, 30 Nov 2004 15:13:43 -0600 From: "Johnson, Teri" Subject: [Histonet] Paraffin processing explant cultures To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" Is anybody doing paraffin processing on explant cultures and can give me some tips on increasing my chances for success? The culture plate inserts are Millicell polycarbonate and my question is whether they can survive tissue processing and paraffin sectioning. If not, what are your recommendations for doing these? Thanks in advance! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org ------------------------------ Message: 14 Date: Tue, 30 Nov 2004 22:59:33 +0100 From: "Gudrun Lang" Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions To: "Histonetliste" Message-ID: <003a01c4d727$e077a3d0$eeeea8c0@server> Content-Type: text/plain; charset="iso-8859-1" Using lithiumcarbonat is the way we do it with staining frozen sections. In the HE-stainer we have running tapwater. And the sections are nice. After haemalaun in special stains we let the slides in tapwater for 3-5 min. Gudrun Lang ----- Original Message ----- From: "Jackie M O'Connor" To: "Johnson, Teri" Cc: "Histonet" ; Sent: Tuesday, November 30, 2004 8:08 PM Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions > Anyone remember using saturated lithium carbonate? I could probably use a > little lithium right now . . . . . > > Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just > bringing the slides BACK to a neutral pH after treating them with acid > which makes them purple-ish? I like the ammonia because it 'seems' to > make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to > make them "blah".(Perception is in the eye of the beholder) Running tap > water will make the same miracle happen, but it takes longer. > > > > > > > "Johnson, Teri" > Sent by: histonet-bounces@lists.utsouthwestern.edu > 11/30/2004 12:51 PM > > > To: "Histonet" > cc: > Subject: [Histonet] Re: Ammonia water pH and other bluing solutions > > > Way back in the day, Brigati was using PBS on his immunostainer to blue > after the hematoxylin counterstain. I have since used PBS to blue my > slides instead of ammonium hydroxide/water. It's cheap, and very easy > on the sections, and I have plenty of it on hand. > > > Teri Johnson > Managing Director Histology Facility > Stowers Institute for Medical Research > 1000 E. 50th St. > Kansas City, Missouri 64110 > tjj@stowers-institute.org > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 15 Date: Tue, 30 Nov 2004 13:59:19 -0800 From: "Luck, Greg D." Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? To: 'Richard Cartun' , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Richard, Would you further define or point me towards a regulatory definition of "Dangerous Goods". Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 8:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Tue, 30 Nov 2004 13:59:42 -0800 From: Fredrik Johansson Subject: RE: [Histonet] Counterstain for Von Kossa To: histonet@lists.utsouthwestern.edu Message-ID: <239E252E-431B-11D9-94FE-000A95F0ED90@u.washington.edu> Content-Type: text/plain; charset=WINDOWS-1252; format=flowed Hi ? Thanks for all the answers about counterstain with Von Kossa. ? It seemed like most of you preferred nuclear fast red. So I'm going to try that. ? Thanks ? Fredrik Johansson Dept. of Pathology UW, Seattle ------------------------------ Message: 17 Date: Tue, 30 Nov 2004 17:02:35 -0500 From: "Tan, MinHan" Subject: [Histonet] Phosphorylated-met antibodies To: Message-ID: Content-Type: text/plain; charset="us-ascii" Good afternoon, We are thinking of staining for phosphorylated-met; while there are a number of commercial antibodies out there, it seems that they have not been tested out on paraffin embedded tissue yet. If anyone has tried staining for this antigen, I'd appreciate any pointers you have. Min-Han This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient(s) please contact the sender by reply email and destroy all copies of the original message. Thank you. ------------------------------ Message: 18 Date: Tue, 30 Nov 2004 15:02:30 -0700 From: "Elizabeth Chlipala" Subject: RE: [Histonet] Paraffin processing explant cultures To: "'Johnson, Teri'" , "'Histonet'" Message-ID: <000001c4d728$49c9c480$76d48a80@AMY> Content-Type: text/plain; charset="us-ascii" Teri I have processed explant cultures with the Millipore MillicellT CM which is a polytetrafluoroethylene based polymer. I process these as any other tissue. Millipore has a great web page in which you can ask questions. I would give them a call about the particular filter that you are using. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Tuesday, November 30, 2004 2:14 PM To: Histonet Subject: [Histonet] Paraffin processing explant cultures Is anybody doing paraffin processing on explant cultures and can give me some tips on increasing my chances for success? The culture plate inserts are Millicell polycarbonate and my question is whether they can survive tissue processing and paraffin sectioning. If not, what are your recommendations for doing these? Thanks in advance! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Tue, 30 Nov 2004 22:24:50 +0000 From: nperson211@comcast.net Subject: [Histonet] CD 133 To: histonet@lists.utsouthwestern.edu Message-ID: <113020042224.12979.41ACF331000E7125000032B32207001641CECECD02019C9D0A9F02@c omcast.net> Content-Type: text/plain Netters, Is anyone using an antibody against CD 133 on FFPE tissues? If so, what have you used for a positive control, that could also provide cells to use as a positive control for Flow? I would really appreciate any information anyone would be willing to share. Thanks, Nancy Lemke Hermelin Brain Tumor Center Henry Ford Hospital Detroit ------------------------------ Message: 20 Date: Tue, 30 Nov 2004 17:01:52 -0600 From: "Johnson, Teri" Subject: RE: [Histonet] Paraffin processing explant cultures To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" Liz is a genius. I called Millipore and their customer service guy, Rich, told me I could paraffin process and section these with no problems. Why didn't *I* think of doing this? Sheesh... Teri -----Original Message----- From: Elizabeth Chlipala [mailto:liz@premierlab.com] Sent: Tuesday, November 30, 2004 4:03 PM To: Johnson, Teri; 'Histonet' Subject: RE: [Histonet] Paraffin processing explant cultures Teri I have processed explant cultures with the Millipore MillicellT CM which is a polytetrafluoroethylene based polymer. I process these as any other tissue. Millipore has a great web page in which you can ask questions. I would give them a call about the particular filter that you are using. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Tuesday, November 30, 2004 2:14 PM To: Histonet Subject: [Histonet] Paraffin processing explant cultures Is anybody doing paraffin processing on explant cultures and can give me some tips on increasing my chances for success? The culture plate inserts are Millicell polycarbonate and my question is whether they can survive tissue processing and paraffin sectioning. If not, what are your recommendations for doing these? Thanks in advance! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: Tue, 30 Nov 2004 15:29:32 -0800 From: "Robyn Vazquez" Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? To: LuckG@empirehealth.org, Rcartun@harthosp.org, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Richard, They are a regulated class 6.2, it is a substance known or reasonalbly expected to contain pathogens. If you ship dangerous goods you will need a shipper's declaration which is required. Robyn OHSU >>> "Luck, Greg D." 11/30/2004 1:59:19 PM >>> Richard, Would you further define or point me towards a regulatory definition of "Dangerous Goods". Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 8:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 22 Date: Tue, 30 Nov 2004 19:13:50 -0500 (EST) From: "SMITH,REBEKAH FELICIA" Subject: Re: [Histonet] Ammonia water pH To: "Jackie M O'Connor" , Tim Webster Cc: histonet@lists.utsouthwestern.edu Message-ID: <934672373.1101860030245.JavaMail.osg@osgjas02.cns.ufl.edu> Content-Type: text/plain; format=flowed; charset=us-ascii I agree with Jackie, I use a Coplin jar with distilled water and about 3 drops of 30% ammonium hydroxide, and then rinse in tap water twice. i've never had problems and it produces a rather pretty blue for me. Rebekah Smith On Tue Nov 30 10:35:55 EST 2004, Jackie M O'Connor wrote: > Maybe I'm being extremely simple-minded - but I just fill up a > 200 ml staining jar with tap H20, and add a couple-three drops of > 28%. When it doesn't smell like ammonia anymore, I make new > stuff. How's that grab ya for QA? "Tim Webster" > > Sent by: histonet-bounces@lists.utsouthwestern.edu > 11/30/2004 09:30 AM > > To: > cc: Subject: [Histonet] Ammonia water pH > > > Hi All, > > I'm curious to know what people's procedures are for making and > using ammonia water as a bluing agent in regressive H&E's. > Currently, our methodology is to make the ammonia water up daily > from a 28% stock bottle, and the pH is about 11. My feeling is > that 11 is way to high and that 9-10 is plenty alkaline to blue > sections adequately. Additionally, I'm concerned about strong > alkaline solution increasing the likelihood of sections washing > off or wrinkling. Sheehan & Hrapchak call for a "weak" alkaline > solution in "Theory and Practice of Histotechnology". Also, I can > see no reason not to make a working solution weekly/monthly and > simply empty and refill the Ammonia water station. The pH > doesn't appear to drop off over time. > > Thanks for your time, and have a great week! Tim > > Tim Webster > Histology Specialist > Northwestern Medical Center > 133 Fairfield Street, > St Albans, VT 05478 > (802) 524-1070 (x4349) > 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. >> If you should >> receive > this message in error, please delete it and notify the sender. > 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 > > -- SMITH,REBEKAH FELICIA "You are a child of the universe, no less than the trees and the stars You have a right to be here and whether or not it is clear to you, no doubt the universe is unfolding as it should. Therefore be at peace with G-d, whatever you conceive Him to be. And whatever your labors and aspirations,in the noisy confusion of life, keep peace in your soul.-Max Ehrmann,"Desiderata" ------------------------------ Message: 23 Date: Tue, 30 Nov 2004 16:17:46 -0800 From: Jennifer MacDonald Subject: Re: [Histonet] Paraffin blocks - Dangerous goods? To: "Richard Cartun" Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Advance for Medical Laboratory Professionals had a good article in the November 1, 2004 issue. It is under the Q&A section, titled: Standards for Shipping Infectious Agents. Check www.advanceweb.com to check the archives. If you can't find it, I can fax you a copy. There is also an old article titled "Safe Specimen Packaging Begins with You. It is from the November 12, 2001 issue. Jennifer MacDonald Mt. San Antonio College "Richard Cartun" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2004 08:12 AM To cc Subject [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 24 Date: Tue, 30 Nov 2004 19:26:06 EST From: RCHIOVETTI@aol.com Subject: Re: [Histonet] gliut/isopropyl alcohol To: BMolinari@heart.thi.tmc.edu, histonet@lists.utsouthwestern.edu Message-ID: <1cc.2d22ffb1.2ede699e@aol.com> Content-Type: text/plain; charset="US-ASCII" In a message dated 11/30/2004 12:12:08 PM US Mountain Standard Time, BMolinari@heart.thi.tmc.edu writes: I received samples that have been fixed in a 1%glut / 20% isopropyl alcohol solution for paraffin processing. I have never heard of this fixative, I know alcoholic formalin but never alcoholic glutaraldehyde. Are any of you familiar with this fixative? Thank you. Betsy Molinari HT (ASCP) Hi Betsy, Is the specimen perhaps a heart valve made from pig tissue?? The only time I've heard of alcoholic glutaraldehyde is for pretreating pig valves before they're implanted in humans. There's some kind of relationship between alcohol and glut, and preventing calcification of the valve. Can't remember the details right now, but you have to treat the valve with one of those reagents before using the other. Just a thought... Cheers, Bob Chiovetti ------------------------------ Message: 25 Date: Tue, 30 Nov 2004 17:28:20 -0800 From: "James Watson" Subject: [Histonet] Part time Histology position in San Diego Ca. To: Message-ID: Content-Type: text/plain; charset="US-ASCII" GNF in San Diego has a part time position available. Hours are flexible, 20 hours per week. Please e-mail resume or call me at the number below. The position requires ASCP HT certification. BS, AA degree, H.S. with completion of certified Histology Program with a minimum of 3 years of experience, or equivalent training with a minimum of 6 years of experience. Must be experienced with frozen sectioning, tissue processing and embedding, paraffin sectioning, H&E and special staining, and immunohistochemistry. Animal tissue trimming and identification is preferred. James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org ------------------------------ Message: 26 Date: Wed, 1 Dec 2004 08:18:41 -0000 From: Kemlo Rogerson Subject: RE: [Histonet] Re: Ammonia water pH and other bluing solutions[Sc anned] To: 'Jackie M O'Connor' , "Johnson, Teri" Cc: Histonet , histonet-bounces@lists.utsouthwestern.edu Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B54C@bhrv-nt-11.bhrv.nwest.nhs.uk> Content-Type: text/plain I think you are wrong, so I'll correct you, as you requested. I thought the 'blue' state of haematin was, like litmus, the effect an alkaline condition has on the siderphilic dye. Red, like litmus, denoted acidic conditions. Damned if I know what colour (with the 'u') haematin goes when neutral (bluey/ red?) Running tap water takes longer, I concede, but good things are worth waiting for and it's easier to make up; we rush too much I find. TIP: Never use London water, been through too many kidneys! -----Original Message----- From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: 30 November 2004 19:09 To: Johnson, Teri Cc: Histonet; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions[Scanned] Anyone remember using saturated lithium carbonate? I could probably use a little lithium right now . . . . . Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just bringing the slides BACK to a neutral pH after treating them with acid which makes them purple-ish? I like the ammonia because it 'seems' to make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to make them "blah".(Perception is in the eye of the beholder) Running tap water will make the same miracle happen, but it takes longer. "Johnson, Teri" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/30/2004 12:51 PM To: "Histonet" cc: Subject: [Histonet] Re: Ammonia water pH and other bluing solutions Way back in the day, Brigati was using PBS on his immunostainer to blue after the hematoxylin counterstain. I have since used PBS to blue my slides instead of ammonium hydroxide/water. It's cheap, and very easy on the sections, and I have plenty of it on hand. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 27 Date: Wed, 1 Dec 2004 08:25:23 -0000 From: Kemlo Rogerson Subject: RE: [Histonet] gliut/isopropyl alcohol[Scanned] To: histonet@lists.utsouthwestern.edu Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B54D@bhrv-nt-11.bhrv.nwest.nhs.uk> Content-Type: text/plain Expect the tissue is rather brittle, used to be used for electromicroscopy I think. Wonder if you could secondary fix? Can't aldehyde/ alcohol fixation be 'washed out' then the tissue refixed in something more conventional? -----Original Message----- From: Molinari, Betsy [mailto:BMolinari@heart.thi.tmc.edu] Sent: 30 November 2004 19:08 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] gliut/isopropyl alcohol[Scanned] Hi histonetters, I received samples that have been fixed in a 1%glut / 20% isopropyl alcohol solution for paraffin processing. I have never heard of this fixative, I know alcoholic formalin but never alcoholic glutaraldehyde. Are any of you familiar with this fixative? Thank you. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 28 Date: Wed, 1 Dec 2004 06:18:48 -0600 From: "Molinari, Betsy" Subject: RE: [Histonet] gliut/isopropyl alcohol To: , Message-ID: Content-Type: text/plain; charset="us-ascii" Hi Bob, Thanks for your input. I may ask a couple of the surgery staff here if they are familiar with that technique. A point of interest, these are valves. I loaded them last night starting in 70% ETOH, I am curious to see how they turned out and how much effect it will have on the B&B and PTAH I will be running on them Thanks again, Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology Houston, TX 77030 832-355-6524 -----Original Message----- From: RCHIOVETTI@aol.com [mailto:RCHIOVETTI@aol.com] Sent: Tuesday, November 30, 2004 6:26 PM To: Molinari, Betsy; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] gliut/isopropyl alcohol In a message dated 11/30/2004 12:12:08 PM US Mountain Standard Time, BMolinari@heart.thi.tmc.edu writes: I received samples that have been fixed in a 1%glut / 20% isopropyl alcohol solution for paraffin processing. I have never heard of this fixative, I know alcoholic formalin but never alcoholic glutaraldehyde. Are any of you familiar with this fixative? Thank you. Betsy Molinari HT (ASCP) Hi Betsy, Is the specimen perhaps a heart valve made from pig tissue?? The only time I've heard of alcoholic glutaraldehyde is for pretreating pig valves before they're implanted in humans. There's some kind of relationship between alcohol and glut, and preventing calcification of the valve. Can't remember the details right now, but you have to treat the valve with one of those reagents before using the other. Just a thought... Cheers, Bob Chiovetti ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 13, Issue 1 *************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Dec 2 09:12:16 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Autofluorescence problem in olfactory epithelia cryosections In-Reply-To: <963396644@web.de> References: <963396644@web.de> Message-ID: <6.0.0.22.1.20041202080450.01b298e0@gemini.msu.montana.edu> Dear Tobias, This has been discussed at great length, even recently, on Histonet. Check the Histonet Archives for even more information on this ongoing annoying problem. Also, I am attaching (privately) a review of autofluorescence that may help you understand some of what you are experiencing. The article also has a way to block autofluorescence, although I have never tried it. Anytime you fix tissues with paraformaldehyde or even formalin, autofluorescence increases due to aldehyde fixation, something that has been observed for many years and with many publications. Some people, as in our lab, often use autofluorescence like a "counterstain" or could we say "counterfluorescence" to contrast with the fluorophore you are using. Blocking autofluorescence is difficult at best, there is an publication found in J of Histochemistry and Cytochemistry that may help you. It is always possible to do a colored chromogen with ISH instead of FISH if this is unsolvable. You did not say what your fluorophore is, FITC? Expect an attachment after this message. At 06:54 AM 12/2/2004, you wrote: >Hello, > >I've got a serious autoflourescence problem when trying to perform >ISH on mouse head cryosections. > >After ISH process I can see a bright red autoflourescence in large >parts of my tissue even without any antibodies on it. >First I thought it might be due to blocking solution containing Horse serum, >but I test blocked my slides and there was almost no autoflourescence at all. >So were the untreated slide ! > >Right now, I don't have a single clue which step of my ISH might cause >the red autofluorescence but it would be great if anyone of you had a good >advice for me. Please, let me know ! > >The mouse has been killed, decapitated, parts of the nose were >removed and fixed in 4%PFA for 3h. After that the tissue was >incubated at 4?C in 30% succrose until the tissue sank to the >bottom of the tube and was than shock frozen and embedded in >mounting medium. Slices are 10?m thick. > >Thank you for your help. > >-Tobias- > > >________________________________________________________________ >Verschicken Sie romantische, coole und witzige Bilder per SMS! >Jetzt neu bei WEB.DE FreeMail: http://freemail.web.de/?mc=021193 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From corinnewillmott <@t> fusemail.com Thu Dec 2 09:33:46 2004 From: corinnewillmott <@t> fusemail.com (Corinne Willmott) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Masson's Trichrome on frozen heart sections Message-ID: <4728.129.67.5.57.1102001626.fusewebmail-25600@www.fusemail.com> Can anyone tell me if it is possible to use Masson's Trichrome stain on frozen cardaic sections and if so is there a recommended protocol? I am new to histology and all the references I've found so far refer to paraffin embedded sections. Thanks Corinne Willmott Stem Cell Research National Blood Service John Radcliffe Hospital Headington Oxford From pruegg <@t> ihctech.net Thu Dec 2 10:33:32 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] post for AZ histo school Message-ID: <200412021633.iB2GXUKr014023@chip.viawest.net> the histology school here in Tucson is expanding rapidly and is in urgent need of usable manual microtomes. Can you put out a message on the Histonet for anyone who is trying to get rid of one to contact me at 800-227-2155, or emacrea@ventanamed.com or Ann Christensen at Pima Community College, Ann.Christensen@pima.edu . The donor can have a tax credit form filled out so they can take the write off as a charitable donation. The school will take anything that is operational. Thanks. Ethel From northma <@t> ohsu.edu Thu Dec 2 10:39:44 2004 From: northma <@t> ohsu.edu (Mary North) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Destaining DifQuik Message-ID: Has anyone tried to destain DifQuik-stained cytology preps so that it can be restained with PAP? Mary North, HT(ASCP), HTL OHSU Portland, OR From juan.gutierrez <@t> christushealth.org Thu Dec 2 10:48:32 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Masson's Trichrome on frozen heart sections Message-ID: We use Gomori's trichrome on frozen sections. It's a one step procedure that gives you a faster answer. Are you able to try alternatives to MTS? Good luck. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: Corinne Willmott [mailto:corinnewillmott@fusemail.com] Sent: Thursday, December 02, 2004 9:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Masson's Trichrome on frozen heart sections Can anyone tell me if it is possible to use Masson's Trichrome stain on frozen cardaic sections and if so is there a recommended protocol? I am new to histology and all the references I've found so far refer to paraffin embedded sections. Thanks Corinne Willmott Stem Cell Research National Blood Service John Radcliffe Hospital Headington Oxford _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From STEGTM <@t> samcstl.org Thu Dec 2 11:07:28 2004 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] M.tuberculosis in formalin-fixed tissue Message-ID: Say y'all. I have a copy of CAP Today in front of me, last months'. There's an article on pg118 that says somewhere around 10% of cultured lung tissue, fixed in formalin, grew M.tuberculosis. We all assume the risk when cutting frozens, and the subsequent decontamination procedures; has anyone really thought about this? Have you heard anything else about such? Does anyone out there assume that this fixed tissue is still viable (we assume that it is a mere capsular ghost of itself). I'd be interested to hear how you deal with this type of tissue; precautions or not...... Terre From gcallis <@t> montana.edu Thu Dec 2 11:20:19 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] fixation for RE: Masson's Trichrome on frozen heart sections In-Reply-To: References: Message-ID: <6.0.0.22.1.20041202101540.01b2baa0@gemini.msu.montana.edu> Juan, How do you fix fresh tissue frozen sections? Bouins or NBF? and fixation times? If we were to do this stain, our tissues are NEVER prefixed before snap freezing. A one step staining procedure makes a lot of sense when handling more fragile frozen sections. I know one substitute light green with aniline blue, or vice versa - what ever one likes best. This was discussed just a few months ago on Histonet. At 09:48 AM 12/2/2004, you wrote: >We use Gomori's trichrome on frozen sections. It's a one step procedure >that gives you a faster answer. Are you able to try alternatives to MTS? >Good luck. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From HornHV <@t> archildrens.org Thu Dec 2 12:08:14 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] New JCAHO PI Standard Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B9BD@EMAIL.archildrens.org> Our pathologists do this. All of our cases are assigned a code and a report is pulled up every month using the codes for review by the surgical affair committee. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Nita Searcy Sent: Thursday, December 02, 2004 7:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New JCAHO PI Standard There is a new standard that states: "An analysis is performed for all major discrepancies between preoperative and postoperative (including pathologic) diagnoses." How are institutions complying ? In my past lives there were "Tissue Committees" that reviewed "normal" tissues but this standard , in my opinion, would not be able to be handled in the pathology lab. We consider ourselves lucky if we get any preoperative diagnosis on our requisitions received from surgery. Perhaps some kind of chart review on all surgeries? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 ------------------------------------------------------------------------------ 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 liz <@t> premierlab.com Thu Dec 2 12:14:47 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] immunohistochemical marker for osteoblasts Message-ID: <000001c4d89a$ce98a550$76d48a80@AMY> Is anyone aware of a Immunohistochemical marker that is specific for osteoblasts? I have seen references on BAP (bone alkaline phosphatase) and some other clones such as 2D3 and 2H10, but I have been unable to find a source. Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From funderwood <@t> mcohio.org Thu Dec 2 12:35:22 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:22 2005 Subject: [BULK] - RE: [Histonet] static electricity Message-ID: Static Guard (brand name) aerosol spray works well. Spray a small amount on your knife clamp and other "sticky" microtome surfaces. >>> 12/02/04 12:26AM >>> Hello, I have the same problem during winter time - and in summer time it is the opposite: it is "common accepted" that in July/August there is hardly any staining possible, because the huminidy goes immediately to the alcohol and then the xylene is milky and mounting not proper. When I have been working in Germany for 10 years I never has this effect, but since I work in Finland I am fighting with it every summer. So, if anyone has a solution to the humidity problem, please share it with me:) Tina -----Original Message----- From: Pereira, Laurie [mailto:Laurie.Pereira@sdcounty.ca.gov] Sent: Thursday, December 02, 2004 1:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] static electricity Hey all, I'm new at Histology and am having a problem with static electricity. During the winter, everything tends to stick to me. I've heard that a humidifier works pretty well but our laboratory is pretty large. Will a humidifier work in such a large space? Does anyone have any hints on how to deal with the wonders of static electricity? Any information would be extremely helpful! Thanks Frustrated _______________________________________________ Histonet mailing list Histonet@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 hessd101 <@t> comcast.net Thu Dec 2 12:55:55 2004 From: hessd101 <@t> comcast.net (hessd101@comcast.net) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] X-gal and pap pens? Message-ID: <120220041855.7738.41AF653B000591D600001E3A2200761438CECFCE0B9C9C0A08@comcast.net> I've searched far and wide for a solution to this problem, and I hope the experts can finally help me. Our lab performs x-gal staining on tissue sections. Prior to 6 months ago, we used Kiyota's Super Pap Pen (which put down an excellent, CLEAR barrier). Our last pen ran (mostly) dry, and we tried to order new pens from Kiyota. Apparently, their clear pens were discontinued because of toxicity, and now the only pen available from them is the Super HT Pap Pen. The new pens put down a flimsy, blue-green colored barrier. The trouble is this - when X-gal staining, some component of the stain reacts with the dye in the pap pen producing a grainy red precipitate which discolors the slide, tissue, etc. In addition, an oily reidue floats up on the solution and can stick to tissue while removing it. In a nutshell, the new pens are awful. Does anyone know of a clear pen that is still sold (EMS, zymed all seem to have colored pens), or have experience with a colored pap pen that doesn't react with xgal staining solution? Thanks Darren Hess MD/PhD student, Dept. of Neuroscience University of Pennsylvania School of Medicine From juan.gutierrez <@t> christushealth.org Thu Dec 2 14:05:54 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] RE: fixation for RE: Masson's Trichrome on frozen heart sections Message-ID: After sectioning we put the slide in a coplin jar with alcoholic formalin for 5 minutes, and proceed with the stain. 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: Gayle Callis [mailto:gcallis@montana.edu] Sent: Thursday, December 02, 2004 11:20 AM To: GUTIERREZ, JUAN; Histonet@lists.utsouthwestern.edu Subject: fixation for RE: Masson's Trichrome on frozen heart sections Juan, How do you fix fresh tissue frozen sections? Bouins or NBF? and fixation times? If we were to do this stain, our tissues are NEVER prefixed before snap freezing. A one step staining procedure makes a lot of sense when handling more fragile frozen sections. I know one substitute light green with aniline blue, or vice versa - what ever one likes best. This was discussed just a few months ago on Histonet. At 09:48 AM 12/2/2004, you wrote: >We use Gomori's trichrome on frozen sections. It's a one step procedure >that gives you a faster answer. Are you able to try alternatives to MTS? >Good luck. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From rgrow <@t> bmnet.com Thu Dec 2 15:48:43 2004 From: rgrow <@t> bmnet.com (rgrow@bmnet.com) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] RE: static electricity/humidity problems In-Reply-To: <200412021439.iB2EduoB008704@dns1.bmnet.com> Message-ID: Tina, There have been several good responses regarding static. Try them. Use what works best in your lab. Humidity is a big problem in the southern United States. Air conditioning systems and de-humidifiers can't handle the load in old drafty buildings. We have solved this problem by adding silica gel crystals to our last absolute alcohol and first xylene in the dehydrating and clearing stations. This step has been successfully used here for many years. It also extends the life of the reagents 2x. We order it from: Sigma Aldrich Company, Catalog #: 21,444.2 Silica Gel 2.5 Kg. A word of caution: Do dispense and discard carefully as it is a carcinogen. Good luck. 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: bettina.hutz@orionpharma.com [mailto:bettina.hutz@orionpharma.com] Sent: 02 December 2004 05:26 To: Laurie.Pereira@sdcounty.ca.gov; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] static electricity[Scanned] Hello, I have the same problem during winter time - and in summer time it is the opposite: it is "common accepted" that in July/August there is hardly any staining possible, because the huminidy goes immediately to the alcohol and then the xylene is milky and mounting not proper. When I have been working in Germany for 10 years I never has this effect, but since I work in Finland I am fighting with it every summer. So, if anyone has a solution to the humidity problem, please share it with me:) Tina From gcallis <@t> montana.edu Thu Dec 2 16:26:32 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] X-gal and pap pens? In-Reply-To: <120220041855.7738.41AF653B000591D600001E3A2200761438CECFCE 0B9C9C0A08@comcast.net> References: <120220041855.7738.41AF653B000591D600001E3A2200761438CECFCE0B9C9C0A08@comcast.net> Message-ID: <6.0.0.22.1.20041202150331.01b51298@gemini.msu.montana.edu> Darren, Try the Vector ImmEdge pens, they are very clean, dry nicely, and have long since replaced Kiyota's pens in our lab. Vector sells their pens in package of two, at a price of one pen from other vendors. One thing to do when using the Immedge pen - shake the begolly's out of them to disperse the active marking goo. Their technical rep recommended this be done. I actually vortex mine to insure goo redistribution. It is interesting that Kiyota's Super Pap Pens were declared toxic. I had wondered about this ( after noticing some very strong solvent odors) and asked them about it, but they said they were acceptable, but that fact changed somewhere along the line. DAKO has a clear pen, but be careful when dispensing - if you press toooooo hard, you have a Niagara Falls of goo all over everything. I macerated ( with a pliers!) their hard tip to make it a tidge fuzzier to capture the goo rush and also pulled a tip from a favorite pen and did a trade of hard tip with softer tip. We also let DAKO pen marks dry longer, but I have that luxury of time and with frozen sections before staining and/or fixation. Hopefully DAKO corrected this over-dispensing problem as the goo is good and it is clear. If all else fails, you can buy hydrophobic barrier slides with lines in a circle, squares or rectangles that work quite well. Just mount the section inside lines and do the staining. Erie has a huge selection of these, and we have even had custom hydrophobic barrier slides made for us to eliminate pap pens. Lovely thing is the barrier stays put unlike the pen marking, and the lines we liked were the one that are flatter like the Erie control slides so mounting a coverslip is more flush with slide surface. You can always talk to Erie about their selection. At 11:55 AM 12/2/2004, you wrote: >I've searched far and wide for a solution to this problem, and I hope the >experts can finally help me. Our lab performs x-gal staining on tissue >sections. Prior to 6 months ago, we used Kiyota's Super Pap Pen (which put >down an excellent, CLEAR barrier). Our last pen ran (mostly) dry, and we >tried to order new pens from Kiyota. Apparently, their clear pens were >discontinued because of toxicity, and now the only pen available from them >is the Super HT Pap Pen. The new pens put down a flimsy, blue-green >colored barrier. The trouble is this - when X-gal staining, some component >of the stain reacts with the dye in the pap pen producing a grainy red >precipitate which discolors the slide, tissue, etc. In addition, an oily >reidue floats up on the solution and can stick to tissue while removing >it. In a nutshell, the new pens are awful. Does anyone know of a clear pen >that is still sold (EMS, zymed all seem to have colored pens), or have >experience with a colored pap pen that doesn'! > t react with xgal staining solution? Thanks > >Darren Hess >MD/PhD student, Dept. of Neuroscience >University of Pennsylvania School of Medicine >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From AnthonyH <@t> chw.edu.au Thu Dec 2 16:46:08 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Destaining DifQuik Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E12B@simba.kids> To destain Romanowsky type stains, place in 1% acetic acid in 90% ethanol. If the smears have been air-dried prior to DifQuik staining, then PAP staining will be very dissapointing since the cells will be affected by air-drying. There is no technique known to reverse this. Regards Tony Henwood JP, 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: Mary North [mailto:northma@ohsu.edu] Sent: Friday, 3 December 2004 3:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Destaining DifQuik Has anyone tried to destain DifQuik-stained cytology preps so that it can be restained with PAP? Mary North, HT(ASCP), HTL OHSU Portland, OR _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From WWmn916 <@t> aol.com Thu Dec 2 22:05:01 2004 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] C4D Vendor Message-ID: <88.1affec28.2ee13fed@aol.com> Greetings everyone, Has anyone heard if there is a C4D vendor? Is there anything on the horizon for availability? Next question.....it may be simple, but does HIPAA rules say patient names can not be on slides? Thanks, Deb King, HT(ASCP) Sacramento, CA From bettina.hutz <@t> orionpharma.com Fri Dec 3 01:51:35 2004 From: bettina.hutz <@t> orionpharma.com (bettina.hutz@orionpharma.com) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] RE: static electricity/humidity problems Message-ID: <243E1A79AF5D89438C70513DBADAA4F0019E3FFD@sfies-exchange1.orionnet.org> Hello Renee, this sounds good, I WILL try it:) Thanks a lot, "kiitos" as we say in finnish:) Tina -----Original Message----- From: rgrow@bmnet.com [mailto:rgrow@bmnet.com] Sent: Thursday, December 02, 2004 11:49 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: static electricity/humidity problems Tina, There have been several good responses regarding static. Try them. Use what works best in your lab. Humidity is a big problem in the southern United States. Air conditioning systems and de-humidifiers can't handle the load in old drafty buildings. We have solved this problem by adding silica gel crystals to our last absolute alcohol and first xylene in the dehydrating and clearing stations. This step has been successfully used here for many years. It also extends the life of the reagents 2x. We order it from: Sigma Aldrich Company, Catalog #: 21,444.2 Silica Gel 2.5 Kg. A word of caution: Do dispense and discard carefully as it is a carcinogen. Good luck. 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: bettina.hutz@orionpharma.com [mailto:bettina.hutz@orionpharma.com] Sent: 02 December 2004 05:26 To: Laurie.Pereira@sdcounty.ca.gov; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] static electricity[Scanned] Hello, I have the same problem during winter time - and in summer time it is the opposite: it is "common accepted" that in July/August there is hardly any staining possible, because the huminidy goes immediately to the alcohol and then the xylene is milky and mounting not proper. When I have been working in Germany for 10 years I never has this effect, but since I work in Finland I am fighting with it every summer. So, if anyone has a solution to the humidity problem, please share it with me:) Tina _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Fri Dec 3 02:13:48 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Destaining DifQuik[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B56F@bhrv-nt-11.bhrv.nwest.nhs.uk> Diff-Quick is usually carried out on air-dried preps and PAP on alcohol fixed. You can destain Diff-quick in 1% acid alcohol or just by leaving in alcohol, but the PAP will be hopeless as it was air-dried. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Mary North [mailto:northma@ohsu.edu] Sent: 02 December 2004 16:40 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Destaining DifQuik[Scanned] Has anyone tried to destain DifQuik-stained cytology preps so that it can be restained with PAP? Mary North, HT(ASCP), HTL OHSU Portland, OR _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Fri Dec 3 02:22:12 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Destaining DifQuik[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B571@bhrv-nt-11.bhrv.nwest.nhs.uk> Oops sorry, you already said it! That's the trouble being in the UK, life has passed you by as you sleep. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: 02 December 2004 22:46 To: 'Mary North'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Destaining DifQuik[Scanned] To destain Romanowsky type stains, place in 1% acetic acid in 90% ethanol. If the smears have been air-dried prior to DifQuik staining, then PAP staining will be very dissapointing since the cells will be affected by air-drying. There is no technique known to reverse this. Regards Tony Henwood JP, 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: Mary North [mailto:northma@ohsu.edu] Sent: Friday, 3 December 2004 3:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Destaining DifQuik Has anyone tried to destain DifQuik-stained cytology preps so that it can be restained with PAP? Mary North, HT(ASCP), HTL OHSU Portland, OR _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Fri Dec 3 02:31:14 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] M.tuberculosis in formalin-fixed tissue[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B572@bhrv-nt-11.bhrv.nwest.nhs.uk> No-one replied to this yet? No? Ok. TB lives (hibernates) in bone, that's why, in the old days people were sent to Sanatoriums. They recovered from their TB, by good diet etc., but when they were released and their diet/ health got worse, the bone de-ossified and the TB was released; they then suffered a relapse. I assume that can happen in lung etc., as the TB may be encapsulated in areas of bony spiculues (can't spell that) it is shielded from the fixative. I assume staining, cutting, etc., can release the bacteria from its bony coffin to make the sections infective. I would assume the risk is low, but there all the same. There is an 'explosion' of TB in London, I gather from the News today. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Therersa Stegall [mailto:STEGTM@samcstl.org] Sent: 02 December 2004 17:07 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] M.tuberculosis in formalin-fixed tissue[Scanned] Say y'all. I have a copy of CAP Today in front of me, last months'. There's an article on pg118 that says somewhere around 10% of cultured lung tissue, fixed in formalin, grew M.tuberculosis. We all assume the risk when cutting frozens, and the subsequent decontamination procedures; has anyone really thought about this? Have you heard anything else about such? Does anyone out there assume that this fixed tissue is still viable (we assume that it is a mere capsular ghost of itself). I'd be interested to hear how you deal with this type of tissue; precautions or not...... Terre _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From arvind <@t> nbrc.res.in Fri Dec 3 03:04:53 2004 From: arvind <@t> nbrc.res.in (Arvind Pundir) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] cryosection fetal tissue Message-ID: <7DE0291A47C4BC4D9CDEF65D5017CEF0A704@mail.nbrc.res.in> can any one pls tell how to embed and cut human fetal tissue for immuno as it is too soft to cut thanks in advance Arvind Singh Pundir National Brain Research Centre Near NSG Campus Manesar, Haryana - 122 050, India arvind@nbrc.res.in Tel.: 91-124 ? 233 8922-26 Fax: 91-124 - 233 89 10 / 91-124 - 233 89 28 aNational Brain Research Centre Manesar, Gurgaon DiNational Brain Research Centre Manesar, Gurgaon Dist. Haryana-122 050, India st. Haryana-122 050, India rch Centre Manesar, Gurgaon Dist. Haryana-122 050, India National Brain Research Centre Manesar, Gurgaon Dist. Haryana-122 050, India National Brain Research Centre Manesar, Gurgaon Dist. Haryana-122 050, India National Brain Research Centre Manesar, Gurgaon Dist. Haryana-122 050, India National Brain Research Centre Manesar, Gurgaon Dist. Haryana-122 050, India From lpwenk <@t> sbcglobal.net Fri Dec 3 04:37:58 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Re: Ammonia water pH and other bluing solutions[Scanned] References: <1030B679AD69D6119C3F00080210DD9D01B0B54C@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: <004d01c4d924$29272da0$2329d445@domainnotset.invalid> Let's see if I can explain pH, hematoxylin, red vs. blue, and bluing, and not get myself into a murky mess of chemistry. Hematoxylin solution is actually an aluminum salt and oxidized hematoxylin, collectively known as aluminum-hematein (Al-hematein). In order to stain just the DNA and RNA, the pH of the Al-hematein solution needs to be between 2.2 to 2.9 (most usually around 2.4-2.5). At this lower (acidic) pH, there are a lot of hydrogen ions (H+) in the Al-Hematein solution. These excess H+ bind to the Al-hematein, causing a shift to the red. When the stained tissue is placed in a solution more alkaline than the 2.2-2.9 stain, there are fewer H+ in the staining solution. This removes the H+ from the Al-Hematein. This causes a color shift in the dye to the blue range (i.e., "bluing the slides"). Therefore, bluing agents must have a pH higher than the mid-2.5. The higher the pH (more alkaline), the faster the H+ are removed from the Al-Hematein, and the faster the "bluing" will take place. Bluing at about pH 5.0 can take up to about 10 minutes to "blue" totally (all red nuclei to some red/some blue (= purple looking tissue) to all blue nuclei). At pH 10, it will take 30 seconds. Tap water is more alkaline than the Al-hematein, so it can be used to blue slides. If the tap water's pH is around 7, this should take about 5 minutes. Some lab's water is more alkaline than other labs, so will take less time. The pH of tap water can change depending upon what chemicals the water treatment facility is using that day to purify the water, so using just tap water to blue may have some variability of time. Most labs rely on an alkaline solution (dilute ammonia, dilute lithium carbonate, Scott's tap water) to speed up the bluing processing and to not rely upon the variability of water pH. Hope this helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Kemlo Rogerson" To: "'Jackie M O'Connor'" ; "Johnson, Teri" Cc: "Histonet" ; Sent: Wednesday, December 01, 2004 3:18 AM Subject: RE: [Histonet] Re: Ammonia water pH and other bluing solutions[Scanned] > I think you are wrong, so I'll correct you, as you requested. > > I thought the 'blue' state of haematin was, like litmus, the effect an > alkaline condition has on the siderphilic dye. Red, like litmus, denoted > acidic conditions. Damned if I know what colour (with the 'u') haematin goes > when neutral (bluey/ red?) > > Running tap water takes longer, I concede, but good things are worth waiting > for and it's easier to make up; we rush too much I find. TIP: Never use > London water, been through too many kidneys! > > -----Original Message----- > From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] > Sent: 30 November 2004 19:09 > To: Johnson, Teri > Cc: Histonet; histonet-bounces@lists.utsouthwestern.edu > Subject: Re: [Histonet] Re: Ammonia water pH and other bluing > solutions[Scanned] > > Anyone remember using saturated lithium carbonate? I could probably use a > little lithium right now . . . . . > > Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just > bringing the slides BACK to a neutral pH after treating them with acid > which makes them purple-ish? I like the ammonia because it 'seems' to > make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to > make them "blah".(Perception is in the eye of the beholder) Running tap > water will make the same miracle happen, but it takes longer. > > > > > > > "Johnson, Teri" > Sent by: histonet-bounces@lists.utsouthwestern.edu > 11/30/2004 12:51 PM > > > To: "Histonet" > cc: > Subject: [Histonet] Re: Ammonia water pH and other bluing > solutions > > > Way back in the day, Brigati was using PBS on his immunostainer to blue > after the hematoxylin counterstain. I have since used PBS to blue my > slides instead of ammonium hydroxide/water. It's cheap, and very easy > on the sections, and I have plenty of it on hand. > > > Teri Johnson > Managing Director Histology Facility > Stowers Institute for Medical Research > 1000 E. 50th St. > Kansas City, Missouri 64110 > tjj@stowers-institute.org > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Fri Dec 3 04:47:26 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Re: Ammonia water pH and other bluing solutions[Sc anned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B573@bhrv-nt-11.bhrv.nwest.nhs.uk> Oh I see now, so why are nuclei blue? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] Sent: 03 December 2004 10:38 To: Kemlo Rogerson; 'Jackie M O'Connor'; Johnson, Teri Cc: Histonet Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions[Scanned] Let's see if I can explain pH, hematoxylin, red vs. blue, and bluing, and not get myself into a murky mess of chemistry. Hematoxylin solution is actually an aluminum salt and oxidized hematoxylin, collectively known as aluminum-hematein (Al-hematein). In order to stain just the DNA and RNA, the pH of the Al-hematein solution needs to be between 2.2 to 2.9 (most usually around 2.4-2.5). At this lower (acidic) pH, there are a lot of hydrogen ions (H+) in the Al-Hematein solution. These excess H+ bind to the Al-hematein, causing a shift to the red. When the stained tissue is placed in a solution more alkaline than the 2.2-2.9 stain, there are fewer H+ in the staining solution. This removes the H+ from the Al-Hematein. This causes a color shift in the dye to the blue range (i.e., "bluing the slides"). Therefore, bluing agents must have a pH higher than the mid-2.5. The higher the pH (more alkaline), the faster the H+ are removed from the Al-Hematein, and the faster the "bluing" will take place. Bluing at about pH 5.0 can take up to about 10 minutes to "blue" totally (all red nuclei to some red/some blue (= purple looking tissue) to all blue nuclei). At pH 10, it will take 30 seconds. Tap water is more alkaline than the Al-hematein, so it can be used to blue slides. If the tap water's pH is around 7, this should take about 5 minutes. Some lab's water is more alkaline than other labs, so will take less time. The pH of tap water can change depending upon what chemicals the water treatment facility is using that day to purify the water, so using just tap water to blue may have some variability of time. Most labs rely on an alkaline solution (dilute ammonia, dilute lithium carbonate, Scott's tap water) to speed up the bluing processing and to not rely upon the variability of water pH. Hope this helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Kemlo Rogerson" To: "'Jackie M O'Connor'" ; "Johnson, Teri" Cc: "Histonet" ; Sent: Wednesday, December 01, 2004 3:18 AM Subject: RE: [Histonet] Re: Ammonia water pH and other bluing solutions[Scanned] > I think you are wrong, so I'll correct you, as you requested. > > I thought the 'blue' state of haematin was, like litmus, the effect an > alkaline condition has on the siderphilic dye. Red, like litmus, denoted > acidic conditions. Damned if I know what colour (with the 'u') haematin goes > when neutral (bluey/ red?) > > Running tap water takes longer, I concede, but good things are worth waiting > for and it's easier to make up; we rush too much I find. TIP: Never use > London water, been through too many kidneys! > > -----Original Message----- > From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] > Sent: 30 November 2004 19:09 > To: Johnson, Teri > Cc: Histonet; histonet-bounces@lists.utsouthwestern.edu > Subject: Re: [Histonet] Re: Ammonia water pH and other bluing > solutions[Scanned] > > Anyone remember using saturated lithium carbonate? I could probably use a > little lithium right now . . . . . > > Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just > bringing the slides BACK to a neutral pH after treating them with acid > which makes them purple-ish? I like the ammonia because it 'seems' to > make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to > make them "blah".(Perception is in the eye of the beholder) Running tap > water will make the same miracle happen, but it takes longer. > > > > > > > "Johnson, Teri" > Sent by: histonet-bounces@lists.utsouthwestern.edu > 11/30/2004 12:51 PM > > > To: "Histonet" > cc: > Subject: [Histonet] Re: Ammonia water pH and other bluing > solutions > > > Way back in the day, Brigati was using PBS on his immunostainer to blue > after the hematoxylin counterstain. I have since used PBS to blue my > slides instead of ammonium hydroxide/water. It's cheap, and very easy > on the sections, and I have plenty of it on hand. > > > Teri Johnson > Managing Director Histology Facility > Stowers Institute for Medical Research > 1000 E. 50th St. > Kansas City, Missouri 64110 > tjj@stowers-institute.org > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Dec 3 06:50:37 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] RE: static electricity/humidity problems Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507601@sjhaexc02.sjha.org> And Cardinal has a product called Molecular Sieve - 4494-500ny that works the same way without the carcinogen factor. (At least as far as I know!) Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of bettina.hutz@orionpharma.com Sent: Friday, December 03, 2004 2:52 AM To: rgrow@bmnet.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: static electricity/humidity problems Hello Renee, this sounds good, I WILL try it:) Thanks a lot, "kiitos" as we say in finnish:) Tina -----Original Message----- From: rgrow@bmnet.com [mailto:rgrow@bmnet.com] Sent: Thursday, December 02, 2004 11:49 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: static electricity/humidity problems Tina, There have been several good responses regarding static. Try them. Use what works best in your lab. Humidity is a big problem in the southern United States. Air conditioning systems and de-humidifiers can't handle the load in old drafty buildings. We have solved this problem by adding silica gel crystals to our last absolute alcohol and first xylene in the dehydrating and clearing stations. This step has been successfully used here for many years. It also extends the life of the reagents 2x. We order it from: Sigma Aldrich Company, Catalog #: 21,444.2 Silica Gel 2.5 Kg. A word of caution: Do dispense and discard carefully as it is a carcinogen. Good luck. 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: bettina.hutz@orionpharma.com [mailto:bettina.hutz@orionpharma.com] Sent: 02 December 2004 05:26 To: Laurie.Pereira@sdcounty.ca.gov; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] static electricity[Scanned] Hello, I have the same problem during winter time - and in summer time it is the opposite: it is "common accepted" that in July/August there is hardly any staining possible, because the huminidy goes immediately to the alcohol and then the xylene is milky and mounting not proper. When I have been working in Germany for 10 years I never has this effect, but since I work in Finland I am fighting with it every summer. So, if anyone has a solution to the humidity problem, please share it with me:) Tina _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From juan.gutierrez <@t> christushealth.org Fri Dec 3 07:41:53 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] C4D Vendor Message-ID: Quidel Corporation 265 N. Whisman Rd. Mountain View, CA 94043 (800)524-6318 We use ours for frozen section IHC. 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: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Thursday, December 02, 2004 10:05 PM To: histonet@pathology.swmed.edu Subject: [Histonet] C4D Vendor Greetings everyone, Has anyone heard if there is a C4D vendor? Is there anything on the horizon for availability? Next question.....it may be simple, but does HIPAA rules say patient names can not be on slides? Thanks, Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Fri Dec 3 07:59:49 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] C4D Vendor Message-ID: Biogenesis, (603) 642-8302 Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of WWmn916@aol.com Sent: Thursday, December 02, 2004 10:05 PM To: histonet@pathology.swmed.edu Subject: [Histonet] C4D Vendor Greetings everyone, Has anyone heard if there is a C4D vendor? Is there anything on the horizon for availability? Next question.....it may be simple, but does HIPAA rules say patient names can not be on slides? Thanks, Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peoshel <@t> wisc.edu Fri Dec 3 08:28:22 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] RE: static electricity/humidity problems In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E507601@sjhaexc02.sjha.org> References: <83AACDB0810528418AA106F9AE9B7F7E507601@sjhaexc02.sjha.org> Message-ID: "Cardinal"? Do you have a website for this company? I've been looking for a supplier of molecular sieve that doesn't charge outrageous prices. I assume the numbers are the catalog number for the sieve -- is it 3 - 4 Angstrom? The sieve pores must be that size to trap water molecules. Thanks. Phil >And Cardinal has a product called Molecular Sieve - 4494-500ny that >works the same way without the carcinogen factor. (At least as far >as I know!) >Joyce > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >bettina.hutz@orionpharma.com >Sent: Friday, December 03, 2004 2:52 AM >To: rgrow@bmnet.com; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: static electricity/humidity problems > > >Hello Renee, > >this sounds good, I WILL try it:) >Thanks a lot, "kiitos" as we say in finnish:) >Tina > >-----Original Message----- >From: rgrow@bmnet.com [mailto:rgrow@bmnet.com] >Sent: Thursday, December 02, 2004 11:49 PM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] RE: static electricity/humidity problems > > >Tina, > >There have been several good responses regarding static. Try them. Use >what works best in your lab. > >Humidity is a big problem in the southern United States. Air conditioning >systems and de-humidifiers can't handle the load in old drafty buildings. We >have solved this problem by adding silica gel crystals to our last absolute >alcohol and first xylene in the dehydrating and clearing stations. This step >has been successfully used here for many years. It also extends the life >of the reagents 2x. We order it from: Sigma Aldrich Company, Catalog #: >21,444.2 Silica Gel 2.5 Kg. A word of caution: Do dispense and discard >carefully as it is a carcinogen. > >Good luck. > >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: bettina.hutz@orionpharma.com [mailto:bettina.hutz@orionpharma.com] >Sent: 02 December 2004 05:26 >To: Laurie.Pereira@sdcounty.ca.gov; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] static electricity[Scanned] > >Hello, > >I have the same problem during winter time - and in summer time it is the >opposite: it is "common accepted" that in July/August there is hardly any >staining possible, because the huminidy goes immediately to the alcohol and >then the xylene is milky and mounting not proper. When I have been working >in Germany for 10 years I never has this effect, but since I work in Finland >I am fighting with it every summer. So, if anyone has a solution to the >humidity problem, please share it with me:) > >Tina > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >Confidentiality Notice ** The information contained in this message >may be privileged and is confidential information intended for the >use of the addressee listed above. If you are neither the intended >recipient nor the employee or agent responsible for delivering this >message to the intended recipient, you are hereby notified that any >disclosure, copying, distribution or the taking of any action in >reliance on the contents of this information is strictly prohibited. >If you have received this communication in error, please notify us >immediately by replying to the message and deleting it from your >computer. >Thank you. Saint Josephs Health System, Inc. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From JWEEMS <@t> sjha.org Fri Dec 3 08:34:23 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] RE: static electricity/humidity problems Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507607@sjhaexc02.sjha.org> Cardinal.com Yes, 4 Angstrom Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Philip Oshel Sent: Friday, December 03, 2004 9:28 AM To: Histonet@Pathology.swmed.edu Subject: RE: [Histonet] RE: static electricity/humidity problems "Cardinal"? Do you have a website for this company? I've been looking for a supplier of molecular sieve that doesn't charge outrageous prices. I assume the numbers are the catalog number for the sieve -- is it 3 - 4 Angstrom? The sieve pores must be that size to trap water molecules. Thanks. Phil >And Cardinal has a product called Molecular Sieve - 4494-500ny that >works the same way without the carcinogen factor. (At least as far >as I know!) >Joyce > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >bettina.hutz@orionpharma.com >Sent: Friday, December 03, 2004 2:52 AM >To: rgrow@bmnet.com; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: static electricity/humidity problems > > >Hello Renee, > >this sounds good, I WILL try it:) >Thanks a lot, "kiitos" as we say in finnish:) >Tina > >-----Original Message----- >From: rgrow@bmnet.com [mailto:rgrow@bmnet.com] >Sent: Thursday, December 02, 2004 11:49 PM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] RE: static electricity/humidity problems > > >Tina, > >There have been several good responses regarding static. Try them. Use >what works best in your lab. > >Humidity is a big problem in the southern United States. Air conditioning >systems and de-humidifiers can't handle the load in old drafty buildings. We >have solved this problem by adding silica gel crystals to our last absolute >alcohol and first xylene in the dehydrating and clearing stations. This step >has been successfully used here for many years. It also extends the life >of the reagents 2x. We order it from: Sigma Aldrich Company, Catalog #: >21,444.2 Silica Gel 2.5 Kg. A word of caution: Do dispense and discard >carefully as it is a carcinogen. > >Good luck. > >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: bettina.hutz@orionpharma.com [mailto:bettina.hutz@orionpharma.com] >Sent: 02 December 2004 05:26 >To: Laurie.Pereira@sdcounty.ca.gov; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] static electricity[Scanned] > >Hello, > >I have the same problem during winter time - and in summer time it is the >opposite: it is "common accepted" that in July/August there is hardly any >staining possible, because the huminidy goes immediately to the alcohol and >then the xylene is milky and mounting not proper. When I have been working >in Germany for 10 years I never has this effect, but since I work in Finland >I am fighting with it every summer. So, if anyone has a solution to the >humidity problem, please share it with me:) > >Tina > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >Confidentiality Notice ** The information contained in this message >may be privileged and is confidential information intended for the >use of the addressee listed above. If you are neither the intended >recipient nor the employee or agent responsible for delivering this >message to the intended recipient, you are hereby notified that any >disclosure, copying, distribution or the taking of any action in >reliance on the contents of this information is strictly prohibited. >If you have received this communication in error, please notify us >immediately by replying to the message and deleting it from your >computer. >Thank you. Saint Josephs Health System, Inc. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From JWEEMS <@t> sjha.org Fri Dec 3 08:39:59 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] RE: static electricity/humidity problems Message-ID: <83AACDB0810528418AA106F9AE9B7F7E50760A@sjhaexc02.sjha.org> And I forgot to say that I'm not sure what your price would be. We are on Premier and I'm not sure if the price would be the same. We pay $65ish for 500 gm. j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Weems, Joyce Sent: Friday, December 03, 2004 9:34 AM To: Philip Oshel; Histonet@Pathology.swmed.edu Subject: RE: [Histonet] RE: static electricity/humidity problems Cardinal.com Yes, 4 Angstrom Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Philip Oshel Sent: Friday, December 03, 2004 9:28 AM To: Histonet@Pathology.swmed.edu Subject: RE: [Histonet] RE: static electricity/humidity problems "Cardinal"? Do you have a website for this company? I've been looking for a supplier of molecular sieve that doesn't charge outrageous prices. I assume the numbers are the catalog number for the sieve -- is it 3 - 4 Angstrom? The sieve pores must be that size to trap water molecules. Thanks. Phil >And Cardinal has a product called Molecular Sieve - 4494-500ny that >works the same way without the carcinogen factor. (At least as far >as I know!) >Joyce > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >bettina.hutz@orionpharma.com >Sent: Friday, December 03, 2004 2:52 AM >To: rgrow@bmnet.com; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: static electricity/humidity problems > > >Hello Renee, > >this sounds good, I WILL try it:) >Thanks a lot, "kiitos" as we say in finnish:) >Tina > >-----Original Message----- >From: rgrow@bmnet.com [mailto:rgrow@bmnet.com] >Sent: Thursday, December 02, 2004 11:49 PM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] RE: static electricity/humidity problems > > >Tina, > >There have been several good responses regarding static. Try them. Use >what works best in your lab. > >Humidity is a big problem in the southern United States. Air conditioning >systems and de-humidifiers can't handle the load in old drafty buildings. We >have solved this problem by adding silica gel crystals to our last absolute >alcohol and first xylene in the dehydrating and clearing stations. This step >has been successfully used here for many years. It also extends the life >of the reagents 2x. We order it from: Sigma Aldrich Company, Catalog #: >21,444.2 Silica Gel 2.5 Kg. A word of caution: Do dispense and discard >carefully as it is a carcinogen. > >Good luck. > >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: bettina.hutz@orionpharma.com [mailto:bettina.hutz@orionpharma.com] >Sent: 02 December 2004 05:26 >To: Laurie.Pereira@sdcounty.ca.gov; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] static electricity[Scanned] > >Hello, > >I have the same problem during winter time - and in summer time it is the >opposite: it is "common accepted" that in July/August there is hardly any >staining possible, because the huminidy goes immediately to the alcohol and >then the xylene is milky and mounting not proper. When I have been working >in Germany for 10 years I never has this effect, but since I work in Finland >I am fighting with it every summer. So, if anyone has a solution to the >humidity problem, please share it with me:) > >Tina > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >Confidentiality Notice ** The information contained in this message >may be privileged and is confidential information intended for the >use of the addressee listed above. If you are neither the intended >recipient nor the employee or agent responsible for delivering this >message to the intended recipient, you are hereby notified that any >disclosure, copying, distribution or the taking of any action in >reliance on the contents of this information is strictly prohibited. >If you have received this communication in error, please notify us >immediately by replying to the message and deleting it from your >computer. >Thank you. Saint Josephs Health System, Inc. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From GDawson <@t> Milw.Dynacare.com Fri Dec 3 08:41:46 2004 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] C4D Vendor Message-ID: I use the one from Biogenesis (#2222-8004). It is for IF on frozen tissue. Good Luck, Glen Dawson -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Thursday, December 02, 2004 10:05 PM To: histonet@pathology.swmed.edu Subject: [Histonet] C4D Vendor Greetings everyone, Has anyone heard if there is a C4D vendor? Is there anything on the horizon for availability? Next question.....it may be simple, but does HIPAA rules say patient names can not be on slides? Thanks, Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Thomas.Crowell <@t> biogenidec.com Fri Dec 3 08:57:29 2004 From: Thomas.Crowell <@t> biogenidec.com (Thomas Crowell) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Mouse Paws In-Reply-To: Message-ID: Hello Histonetters, I'm looking for some help or guidance in obtaining good sections from mouse front paws, to include the bones of the wrist and digits, as well as maintaining the surrounding matrix. This is an ongoing problem in our laboratory and we have a difficult time obtaining good sections at this level, the problem appearing to be inadequate infiltration of paraffin into the interior of the paws, leaving the center somewhat mushy & white. The feet are fixed in 10% NBF for up to a week, decalcified in formic acid/sodium citrate for 3 to 5 days, processed on a Shandon Pathcentre for 45 minutes/reagent, 4 changes of paraffin, 45 minutes each, vacum & pressure on the waxes only. Any suggestions regarding fixation, decalcification, or processing enhancements to improve the overall picture would be extremely helpful. Thanks! Tom Crowell BiogenIdec Cambridge, MA From sa.drew <@t> hosp.wisc.edu Fri Dec 3 09:10:57 2004 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] C4D Vendor Message-ID: As my co-worker indicated, we use Biogenesis' C4d. I just thought I'd mention that we have also used it on formalin-fixed, paraffin embedded tissue also...cardiac biopsies. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of WWmn916@aol.com Sent: Thursday, December 02, 2004 10:05 PM To: histonet@pathology.swmed.edu Subject: [Histonet] C4D Vendor Greetings everyone, Has anyone heard if there is a C4D vendor? Is there anything on the horizon for availability? Next question.....it may be simple, but does HIPAA rules say patient names can not be on slides? Thanks, Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jryan <@t> sleh.com Thu Dec 2 12:05:12 2004 From: jryan <@t> sleh.com (John Ryan) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Paraffin blocks Message-ID: I sent an email to IATA ( International Air Transport Association) regarding paraffin blocks that contain tissue not known to be infectious and received a reply that they are not considered dangerous goods. This reply was received from the Marian Terlecki, Manager of Cargo Security and ULD. John P Ryan, HT(ASCP)HTL Assistant Administrative Director Pathology St. Luke's Episcopal Hospital 832-355-2643 "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-request@lists.utsouthwestern.edu 12/02/04 09:17AM >>> 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: Type of hematoxylin stain with your bluing methods? Tell all, please. (Gudrun Lang) 2. Re: Type of hematoxylin stain with your bluing methods? Tell all, please. (Kathleen Roberts) 3. RE: Paraffin blocks - Dangerous goods? (Bauer, Karen) 4. RE: Paraffin blocks - Dangerous goods? (Stacy McLaughlin) 5. RE: Paraffin blocks - Dangerous goods? (Robyn Vazquez) 6. RE: Type of hematoxylin stain with your bluing methods ? Tell all, please. [Scanned] (Robyn Vazquez) 7. RE: Enzyme stains (GUTIERREZ, JUAN) 8. Milestone Microwave Processor (Nita Searcy) 9. RE: Paraffin blocks - Dangerous goods? (Bauer, Karen) 10. Re: Milestone Microwave Processor (Mary Reeves) 11. static electricity (Pereira, Laurie ) 12. RE: static electricity (Weems, Joyce) 13. re:static electricity (Pereira, Laurie ) 14. A good illiustrated book (Shrttmprd@aol.com) 15. RE: Type of hematoxylin stain with your bluing methods ? Tell all, please. [Scanned] (SMITH,REBEKAH FELICIA) 16. RE: static electricity (bettina.hutz@orionpharma.com) 17. Re: Enzyme stains (John Kiernan) 18. RE: static electricity[Scanned] (Kemlo Rogerson) 19. New JCAHO PI Standard (Nita Searcy) 20. Autofluorescence problem in olfactory epithelia cryosections (Tobias M?tzig) 21. Tissue Disposal ? (bliven.laura@marshfieldclinic.org) 22. RE: RE: Autoimmunostainers DAKO vs Ventanna (Dawson, Glen) ---------------------------------------------------------------------- Message: 1 Date: Wed, 1 Dec 2004 19:35:38 +0100 From: "Gudrun Lang" Subject: [Histonet] Re: Type of hematoxylin stain with your bluing methods? Tell all, please. To: "Gayle Callis" , "Histonetliste" Message-ID: <003601c4d7d4$8e3bd160$eeeea8c0@server> Content-Type: text/plain; charset="iso-8859-1" Harris - frozen sections Mayers - HE stainer Gudrun ----- Original Message ----- From: "Gayle Callis" To: "Gudrun Lang" ; Sent: Wednesday, December 01, 2004 4:29 PM Subject: Type of hematoxylin stain with your bluing methods? Tell all, please. > Dear All, > > It would be appreciated that you say which hematoxylin you are using when > you give your bluing method. For Gudrun, are you using Mayers? > > At 02:59 PM 11/30/2004, you wrote: > >Using lithiumcarbonat is the way we do it with staining frozen sections. > >In the HE-stainer we have running tapwater. And the sections are nice. > >After haemalaun in special stains we let the slides in tapwater for 3-5 min. > > > >Gudrun Lang > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > ------------------------------ Message: 2 Date: Wed, 01 Dec 2004 13:55:27 -0500 From: Kathleen Roberts Subject: Re: [Histonet] Type of hematoxylin stain with your bluing methods? Tell all, please. To: Gayle Callis , "'histonet@lists.utsouthwestern.edu'" Message-ID: <41AE139F.20203@rci.rutgers.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed We use Gill's formulation #3 from Fisher Scientific. Kathleen Gayle Callis wrote: > Dear All, > > It would be appreciated that you say which hematoxylin you are using > when you give your bluing method. For Gudrun, are you using Mayers? > > At 02:59 PM 11/30/2004, you wrote: > >> Using lithiumcarbonat is the way we do it with staining frozen sections. >> In the HE-stainer we have running tapwater. And the sections are nice. >> After haemalaun in special stains we let the slides in tapwater for >> 3-5 min. >> >> Gudrun Lang > > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Wed, 1 Dec 2004 12:51:14 -0600 From: "Bauer, Karen" Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? To: "'Richard Cartun'" , "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Richard, The primary container does NOT NEED to be a bio-hazard bag. Sorry, I just used that as a suggestion. I did ask our safety person about the paraffin block being the "primary container", but she thought that a plain (small) bag could be used as the primary container and then a bio-hazard bag as the secondary container, since the secondary container is required to have "bio-hazard" on the outside. I realize that "we" handle blocks without gloves all the time, so I'm not sure what to tell you about that. I guess you'll have to check with your infection control person (I'd hate to open a "can-o-worms" here). The regulations are more for the transporting personnel so they know how to handle certain "packages". (That's just what I think...) Hope this helps... Karen -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Wednesday, December 01, 2004 11:50 AM To: Bauer, Karen L. Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Thank you for the detailed information. One question - If you put the paraffin block into a "Biohazard" bag, doesn't that mean that the person who opens the bag needs to be wearing gloves? We (histotechs, lab aides, secretaries, pathologists, etc.) handle paraffin tissue blocks all the time without wearing gloves. RWC 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 >>> "Bauer, Karen" 12/01/04 12:41PM >>> Our safety person in the lab has just "trained" us all on the proper packaging of specimens in the lab based on the new regulations that are out. Regulations come from the International Civil Aviation Organization (ICAO), the International Air Transport Association (IATA), the USPS, and the Department of Transportation (DOT). Risk groups are ranged from 1 to 4 with 1 being "No to Low Risk" and 4 being "High Risk". Low risk specimens are things like diagnostic serum, whole blood, urine, or tissue samples not known to contain pathogens. Packaging for these specimens need to have a primary container (a bio-hazard bag would be fine for tissue blocks, or maybe the paraffin block itself is the primary container since the tissue is embedded inside it. I'll have to check on that...), then a secondary container (which could be another bio-hazard bag), and then an outer package that is sturdy. This could be a cardboard box or one of those insulated manila envelopes used for fragile items. We do not need to really worry about the outer package, since the couriers that pick up our specimens from the lab have coolers or bio-hazard containers that they put the specimens into. This is considered the "outer package". Our safety person then had everyone who went through the presentation take a short quiz (for competency) and gave us a "Certificate" of being trained. This training will need to be done every 2 years. Everyone in the lab is being trained, since at some point we will all need to know how to ship something out whether it will be Diagnostic or Infectious Specimens (Risk 1 to Risk 4). Karen Bauer HT(ASCP Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 10:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. ------------------------------ Message: 4 Date: Wed, 1 Dec 2004 14:06:02 -0500 From: Stacy McLaughlin Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? To: "'Bauer, Karen'" , 'Richard Cartun' , histonet@lists.utsouthwestern.edu Message-ID: <3D502BBF5356D31184650090275B750D0346C892@mail.cooley-dickinson.org> Content-Type: text/plain Is anyone checking that this requirement is being fulfilled? (CAP, JCAHO, OSHA, etc...) I want to make sure the right thing is being done, but I also would like to know if anyone is watching for that specifically. Thanks, Stacy McLaughlin HT (ASCP) (and Lab Safety Lady) Cooley Dickinson Hospital Northampton,MA -----Original Message----- From: Bauer, Karen [mailto:Bauer.Karen@mayo.edu] Sent: Wednesday, December 01, 2004 12:42 PM To: 'Richard Cartun'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Our safety person in the lab has just "trained" us all on the proper packaging of specimens in the lab based on the new regulations that are out. Regulations come from the International Civil Aviation Organization (ICAO), the International Air Transport Association (IATA), the USPS, and the Department of Transportation (DOT). Risk groups are ranged from 1 to 4 with 1 being "No to Low Risk" and 4 being "High Risk". Low risk specimens are things like diagnostic serum, whole blood, urine, or tissue samples not known to contain pathogens. Packaging for these specimens need to have a primary container (a bio-hazard bag would be fine for tissue blocks, or maybe the paraffin block itself is the primary container since the tissue is embedded inside it. I'll have to check on that...), then a secondary container (which could be another bio-hazard bag), and then an outer package that is sturdy. This could be a cardboard box or one of those insulated manila envelopes used for fragile items. We do not need to really worry about the outer package, since the couriers that pick up our specimens from the lab have coolers or bio-hazard containers that they put the specimens into. This is considered the "outer package". Our safety person then had everyone who went through the presentation take a short quiz (for competency) and gave us a "Certificate" of being trained. This training will need to be done every 2 years. Everyone in the lab is being trained, since at some point we will all need to know how to ship something out whether it will be Diagnostic or Infectious Specimens (Risk 1 to Risk 4). Karen Bauer HT(ASCP Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 10:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet THIS E-MAIL MESSAGE IS INTENDED ONLY FOR THE USE OF THE INDIVIDUAL OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If the reader of this e-mail 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 e-mail message is strictly prohibited. If you have received this e-mail message in error, please immediately notify Cooley Dickinson Healthcare at 413-582-2000 and delete or shred the original message and all copies thereof. Thank you. ------------------------------ Message: 5 Date: Wed, 01 Dec 2004 11:15:09 -0800 From: "Robyn Vazquez" Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? To: LuckG@empirehealth.org, Rcartun@harthosp.org, Mackinnon@holburn.com, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii John, Now that I think about it, I get blocks in the mail occasionally. Maybe it is a regulation that is not known to a doctors office or it is a turn blind eye regulation, because it's not like getting a slab of meat in the mail ;0) Robyn OHSU >>> "John Mackinnon" 12/1/2004 8:20:02 AM >>> I would say that that regulation would be open to interpretation. My interpretation would be that most tissues following fixation do not contain viable pathogens and in most cases it would be reasonable to expect that it does not contain pathogens and is therefore not a biohazard. I know that there are some infectious agents that survive fixation and processing but in most cases we are aware of these because they have been diagnosed (most of the time). These few cases fit the definition and should be handled accordingly. This is just one mans opinion. P.S. I wonder if all of the meat packing plants include a shipper's declaration (E.coli, BSE) with their raw unfixed carcases they ship to the grocery stores and butcher shops. Now there is something to think about. John MacKinnon MLT, ART Laboratory Director Holburn Biomedical Corporation 905-623-1484 mackinnon@holburn.com This transmission may contain information that is privileged, confidential and/or exempt from disclosure under applicable law. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or use of the information contained herein (including any reliance thereon) is STRICTLY PROHIBITED. If you received this transmission in error, please immediately contact the sender and destroy the material in its entirety, whether in electronic or hard copy format. Thank you. -----Original Message----- From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] Sent: November 30, 2004 6:30 PM To: LuckG@empirehealth.org; Rcartun@harthosp.org; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Richard, They are a regulated class 6.2, it is a substance known or reasonalbly expected to contain pathogens. If you ship dangerous goods you will need a shipper's declaration which is required. Robyn OHSU >>> "Luck, Greg D." 11/30/2004 1:59:19 PM >>> Richard, Would you further define or point me towards a regulatory definition of "Dangerous Goods". Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 8:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Wed, 01 Dec 2004 11:16:24 -0800 From: "Robyn Vazquez" Subject: RE: [Histonet] Type of hematoxylin stain with your bluing methods ? Tell all, please. [Scanned] To: Kemlo.Rogerson@elht.nhs.uk, Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Shandon Instant Hematoxylin >>> Kemlo Rogerson 12/1/2004 8:30:30 AM >>> Harris's and Gill's. -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 01 December 2004 15:30 To: Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Type of hematoxylin stain with your bluing methods? Tell all, please. [Scanned] Dear All, It would be appreciated that you say which hematoxylin you are using when you give your bluing method. For Gudrun, are you using Mayers? At 02:59 PM 11/30/2004, you wrote: >Using lithiumcarbonat is the way we do it with staining frozen sections. >In the HE-stainer we have running tapwater. And the sections are nice. >After haemalaun in special stains we let the slides in tapwater for 3-5 min. > >Gudrun Lang Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Wed, 1 Dec 2004 14:00:00 -0600 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] Enzyme stains To: , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Novocastra has a few different antibodies for cathepsin. www.novocastra.co.uk 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: BennettW@pac.dfo-mpo.gc.ca [mailto:BennettW@pac.dfo-mpo.gc.ca] Sent: Wednesday, December 01, 2004 11:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Enzyme stains Hello Histonetters, Does anyone know if there is anything that will stain an enzyme called cathepsin. This is an enzyme that is produced by a myxobilatus sp. parasite (Kudoa thyristes) found in fish. The parasite produces this protealytic enzyme. My preliminary searches haven't revealed too much more than that. Any help would be great. Cheers Bill Bennett Histologist Fisheries and Oceans Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Wed, 01 Dec 2004 14:21:16 -0600 From: "Nita Searcy" Subject: [Histonet] Milestone Microwave Processor To: Message-ID: Content-Type: text/plain; charset="us-ascii" After 18 months in service, we are experiencing paraffin being drawn into the vacuum tube - anyone else having this problem? Can anyone provide help ? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:4-2438 ORG:;Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD ------------------------------ Message: 9 Date: Wed, 1 Dec 2004 14:27:40 -0600 From: "Bauer, Karen" Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? To: "'Stacy McLaughlin'" , "'Bauer, Karen'" , "'Richard Cartun'" , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Stacy, If you go to the CAP TODAY, Oct. 2004 issue on page 101, there is a Q & A about shipping of materials. It is also on the CAP General Checklist # GEN.40522. CAP will be changing their training requirements to every two years instead of the "annual training" that they have on the checklist now. Hope this helps, Karen -----Original Message----- From: Stacy McLaughlin [mailto:Stacy_McLaughlin@cooley-dickinson.org] Sent: Wednesday, December 01, 2004 1:06 PM To: 'Bauer, Karen'; 'Richard Cartun'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Is anyone checking that this requirement is being fulfilled? (CAP, JCAHO, OSHA, etc...) I want to make sure the right thing is being done, but I also would like to know if anyone is watching for that specifically. Thanks, Stacy McLaughlin HT (ASCP) (and Lab Safety Lady) Cooley Dickinson Hospital Northampton,MA -----Original Message----- From: Bauer, Karen [mailto:Bauer.Karen@mayo.edu] Sent: Wednesday, December 01, 2004 12:42 PM To: 'Richard Cartun'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Our safety person in the lab has just "trained" us all on the proper packaging of specimens in the lab based on the new regulations that are out. Regulations come from the International Civil Aviation Organization (ICAO), the International Air Transport Association (IATA), the USPS, and the Department of Transportation (DOT). Risk groups are ranged from 1 to 4 with 1 being "No to Low Risk" and 4 being "High Risk". Low risk specimens are things like diagnostic serum, whole blood, urine, or tissue samples not known to contain pathogens. Packaging for these specimens need to have a primary container (a bio-hazard bag would be fine for tissue blocks, or maybe the paraffin block itself is the primary container since the tissue is embedded inside it. I'll have to check on that...), then a secondary container (which could be another bio-hazard bag), and then an outer package that is sturdy. This could be a cardboard box or one of those insulated manila envelopes used for fragile items. We do not need to really worry about the outer package, since the couriers that pick up our specimens from the lab have coolers or bio-hazard containers that they put the specimens into. This is considered the "outer package". Our safety person then had everyone who went through the presentation take a short quiz (for competency) and gave us a "Certificate" of being trained. This training will need to be done every 2 years. Everyone in the lab is being trained, since at some point we will all need to know how to ship something out whether it will be Diagnostic or Infectious Specimens (Risk 1 to Risk 4). Karen Bauer HT(ASCP Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 10:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet THIS E-MAIL MESSAGE IS INTENDED ONLY FOR THE USE OF THE INDIVIDUAL OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If the reader of this e-mail 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 e-mail message is strictly prohibited. If you have received this e-mail message in error, please immediately notify Cooley Dickinson Healthcare at 413-582-2000 and delete or shred the original message and all copies thereof. Thank you. ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. ------------------------------ +++++CONFIDENTIALITY NOTICE+++++ The information in this e-mail may be confidential and/or privileged. If you are not the intended recipient or an authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments, if any, or the information contained herein is prohibited. If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail from your computer system. Thank you. From Jackie.O'Connor <@t> abbott.com Fri Dec 3 09:18:42 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Overfixed tissues for IHC Message-ID: Happy Friday - I've been asked if tissues that have been in formalin for 5 months are still good for IHC (I don't know what antibodies they want). I'd appreciate your feedback on this one, anyone and everyone. Thanks - Jackie O' From bmcmahill <@t> incytepathology.com Fri Dec 3 09:25:27 2004 From: bmcmahill <@t> incytepathology.com (Bonnie J. McMahill) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] C4D Vendor Message-ID: I have found two vendors for C4d for paraffin, the other one is ARP- American Research Products (617) 489-1120 Bonnie McMahill Histology Supervisor InCyte Pathology Spokane, WA > -----Original Message----- > From: WWmn916@aol.com [SMTP:WWmn916@aol.com] > Sent: Thursday, December 02, 2004 8:05 PM > To: histonet@pathology.swmed.edu > Subject: [Histonet] C4D Vendor > > Greetings everyone, > > Has anyone heard if there is a C4D vendor? Is there anything on the horizon > for availability? > > Next question.....it may be simple, but does HIPAA rules say patient names > can not be on slides? > > Thanks, > Deb King, HT(ASCP) > Sacramento, CA > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Dec 3 09:29:28 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Paraffin blocks Message-ID: <83AACDB0810528418AA106F9AE9B7F7E50760D@sjhaexc02.sjha.org> Thank you John! j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of John Ryan Sent: Thursday, December 02, 2004 1:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks I sent an email to IATA ( International Air Transport Association) regarding paraffin blocks that contain tissue not known to be infectious and received a reply that they are not considered dangerous goods. This reply was received from the Marian Terlecki, Manager of Cargo Security and ULD. John P Ryan, HT(ASCP)HTL Assistant Administrative Director Pathology St. Luke's Episcopal Hospital 832-355-2643 "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-request@lists.utsouthwestern.edu 12/02/04 09:17AM >>> 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: Type of hematoxylin stain with your bluing methods? Tell all, please. (Gudrun Lang) 2. Re: Type of hematoxylin stain with your bluing methods? Tell all, please. (Kathleen Roberts) 3. RE: Paraffin blocks - Dangerous goods? (Bauer, Karen) 4. RE: Paraffin blocks - Dangerous goods? (Stacy McLaughlin) 5. RE: Paraffin blocks - Dangerous goods? (Robyn Vazquez) 6. RE: Type of hematoxylin stain with your bluing methods ? Tell all, please. [Scanned] (Robyn Vazquez) 7. RE: Enzyme stains (GUTIERREZ, JUAN) 8. Milestone Microwave Processor (Nita Searcy) 9. RE: Paraffin blocks - Dangerous goods? (Bauer, Karen) 10. Re: Milestone Microwave Processor (Mary Reeves) 11. static electricity (Pereira, Laurie ) 12. RE: static electricity (Weems, Joyce) 13. re:static electricity (Pereira, Laurie ) 14. A good illiustrated book (Shrttmprd@aol.com) 15. RE: Type of hematoxylin stain with your bluing methods ? Tell all, please. [Scanned] (SMITH,REBEKAH FELICIA) 16. RE: static electricity (bettina.hutz@orionpharma.com) 17. Re: Enzyme stains (John Kiernan) 18. RE: static electricity[Scanned] (Kemlo Rogerson) 19. New JCAHO PI Standard (Nita Searcy) 20. Autofluorescence problem in olfactory epithelia cryosections (Tobias M?tzig) 21. Tissue Disposal ? (bliven.laura@marshfieldclinic.org) 22. RE: RE: Autoimmunostainers DAKO vs Ventanna (Dawson, Glen) ---------------------------------------------------------------------- Message: 1 Date: Wed, 1 Dec 2004 19:35:38 +0100 From: "Gudrun Lang" Subject: [Histonet] Re: Type of hematoxylin stain with your bluing methods? Tell all, please. To: "Gayle Callis" , "Histonetliste" Message-ID: <003601c4d7d4$8e3bd160$eeeea8c0@server> Content-Type: text/plain; charset="iso-8859-1" Harris - frozen sections Mayers - HE stainer Gudrun ----- Original Message ----- From: "Gayle Callis" To: "Gudrun Lang" ; Sent: Wednesday, December 01, 2004 4:29 PM Subject: Type of hematoxylin stain with your bluing methods? Tell all, please. > Dear All, > > It would be appreciated that you say which hematoxylin you are using when > you give your bluing method. For Gudrun, are you using Mayers? > > At 02:59 PM 11/30/2004, you wrote: > >Using lithiumcarbonat is the way we do it with staining frozen sections. > >In the HE-stainer we have running tapwater. And the sections are nice. > >After haemalaun in special stains we let the slides in tapwater for 3-5 min. > > > >Gudrun Lang > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > ------------------------------ Message: 2 Date: Wed, 01 Dec 2004 13:55:27 -0500 From: Kathleen Roberts Subject: Re: [Histonet] Type of hematoxylin stain with your bluing methods? Tell all, please. To: Gayle Callis , "'histonet@lists.utsouthwestern.edu'" Message-ID: <41AE139F.20203@rci.rutgers.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed We use Gill's formulation #3 from Fisher Scientific. Kathleen Gayle Callis wrote: > Dear All, > > It would be appreciated that you say which hematoxylin you are using > when you give your bluing method. For Gudrun, are you using Mayers? > > At 02:59 PM 11/30/2004, you wrote: > >> Using lithiumcarbonat is the way we do it with staining frozen sections. >> In the HE-stainer we have running tapwater. And the sections are nice. >> After haemalaun in special stains we let the slides in tapwater for >> 3-5 min. >> >> Gudrun Lang > > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Wed, 1 Dec 2004 12:51:14 -0600 From: "Bauer, Karen" Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? To: "'Richard Cartun'" , "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Richard, The primary container does NOT NEED to be a bio-hazard bag. Sorry, I just used that as a suggestion. I did ask our safety person about the paraffin block being the "primary container", but she thought that a plain (small) bag could be used as the primary container and then a bio-hazard bag as the secondary container, since the secondary container is required to have "bio-hazard" on the outside. I realize that "we" handle blocks without gloves all the time, so I'm not sure what to tell you about that. I guess you'll have to check with your infection control person (I'd hate to open a "can-o-worms" here). The regulations are more for the transporting personnel so they know how to handle certain "packages". (That's just what I think...) Hope this helps... Karen -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Wednesday, December 01, 2004 11:50 AM To: Bauer, Karen L. Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Thank you for the detailed information. One question - If you put the paraffin block into a "Biohazard" bag, doesn't that mean that the person who opens the bag needs to be wearing gloves? We (histotechs, lab aides, secretaries, pathologists, etc.) handle paraffin tissue blocks all the time without wearing gloves. RWC 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 >>> "Bauer, Karen" 12/01/04 12:41PM >>> Our safety person in the lab has just "trained" us all on the proper packaging of specimens in the lab based on the new regulations that are out. Regulations come from the International Civil Aviation Organization (ICAO), the International Air Transport Association (IATA), the USPS, and the Department of Transportation (DOT). Risk groups are ranged from 1 to 4 with 1 being "No to Low Risk" and 4 being "High Risk". Low risk specimens are things like diagnostic serum, whole blood, urine, or tissue samples not known to contain pathogens. Packaging for these specimens need to have a primary container (a bio-hazard bag would be fine for tissue blocks, or maybe the paraffin block itself is the primary container since the tissue is embedded inside it. I'll have to check on that...), then a secondary container (which could be another bio-hazard bag), and then an outer package that is sturdy. This could be a cardboard box or one of those insulated manila envelopes used for fragile items. We do not need to really worry about the outer package, since the couriers that pick up our specimens from the lab have coolers or bio-hazard containers that they put the specimens into. This is considered the "outer package". Our safety person then had everyone who went through the presentation take a short quiz (for competency) and gave us a "Certificate" of being trained. This training will need to be done every 2 years. Everyone in the lab is being trained, since at some point we will all need to know how to ship something out whether it will be Diagnostic or Infectious Specimens (Risk 1 to Risk 4). Karen Bauer HT(ASCP Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 10:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. ------------------------------ Message: 4 Date: Wed, 1 Dec 2004 14:06:02 -0500 From: Stacy McLaughlin Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? To: "'Bauer, Karen'" , 'Richard Cartun' , histonet@lists.utsouthwestern.edu Message-ID: <3D502BBF5356D31184650090275B750D0346C892@mail.cooley-dickinson.org> Content-Type: text/plain Is anyone checking that this requirement is being fulfilled? (CAP, JCAHO, OSHA, etc...) I want to make sure the right thing is being done, but I also would like to know if anyone is watching for that specifically. Thanks, Stacy McLaughlin HT (ASCP) (and Lab Safety Lady) Cooley Dickinson Hospital Northampton,MA -----Original Message----- From: Bauer, Karen [mailto:Bauer.Karen@mayo.edu] Sent: Wednesday, December 01, 2004 12:42 PM To: 'Richard Cartun'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Our safety person in the lab has just "trained" us all on the proper packaging of specimens in the lab based on the new regulations that are out. Regulations come from the International Civil Aviation Organization (ICAO), the International Air Transport Association (IATA), the USPS, and the Department of Transportation (DOT). Risk groups are ranged from 1 to 4 with 1 being "No to Low Risk" and 4 being "High Risk". Low risk specimens are things like diagnostic serum, whole blood, urine, or tissue samples not known to contain pathogens. Packaging for these specimens need to have a primary container (a bio-hazard bag would be fine for tissue blocks, or maybe the paraffin block itself is the primary container since the tissue is embedded inside it. I'll have to check on that...), then a secondary container (which could be another bio-hazard bag), and then an outer package that is sturdy. This could be a cardboard box or one of those insulated manila envelopes used for fragile items. We do not need to really worry about the outer package, since the couriers that pick up our specimens from the lab have coolers or bio-hazard containers that they put the specimens into. This is considered the "outer package". Our safety person then had everyone who went through the presentation take a short quiz (for competency) and gave us a "Certificate" of being trained. This training will need to be done every 2 years. Everyone in the lab is being trained, since at some point we will all need to know how to ship something out whether it will be Diagnostic or Infectious Specimens (Risk 1 to Risk 4). Karen Bauer HT(ASCP Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 10:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet THIS E-MAIL MESSAGE IS INTENDED ONLY FOR THE USE OF THE INDIVIDUAL OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If the reader of this e-mail 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 e-mail message is strictly prohibited. If you have received this e-mail message in error, please immediately notify Cooley Dickinson Healthcare at 413-582-2000 and delete or shred the original message and all copies thereof. Thank you. ------------------------------ Message: 5 Date: Wed, 01 Dec 2004 11:15:09 -0800 From: "Robyn Vazquez" Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? To: LuckG@empirehealth.org, Rcartun@harthosp.org, Mackinnon@holburn.com, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii John, Now that I think about it, I get blocks in the mail occasionally. Maybe it is a regulation that is not known to a doctors office or it is a turn blind eye regulation, because it's not like getting a slab of meat in the mail ;0) Robyn OHSU >>> "John Mackinnon" 12/1/2004 8:20:02 AM >>> I would say that that regulation would be open to interpretation. My interpretation would be that most tissues following fixation do not contain viable pathogens and in most cases it would be reasonable to expect that it does not contain pathogens and is therefore not a biohazard. I know that there are some infectious agents that survive fixation and processing but in most cases we are aware of these because they have been diagnosed (most of the time). These few cases fit the definition and should be handled accordingly. This is just one mans opinion. P.S. I wonder if all of the meat packing plants include a shipper's declaration (E.coli, BSE) with their raw unfixed carcases they ship to the grocery stores and butcher shops. Now there is something to think about. John MacKinnon MLT, ART Laboratory Director Holburn Biomedical Corporation 905-623-1484 mackinnon@holburn.com This transmission may contain information that is privileged, confidential and/or exempt from disclosure under applicable law. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or use of the information contained herein (including any reliance thereon) is STRICTLY PROHIBITED. If you received this transmission in error, please immediately contact the sender and destroy the material in its entirety, whether in electronic or hard copy format. Thank you. -----Original Message----- From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] Sent: November 30, 2004 6:30 PM To: LuckG@empirehealth.org; Rcartun@harthosp.org; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Richard, They are a regulated class 6.2, it is a substance known or reasonalbly expected to contain pathogens. If you ship dangerous goods you will need a shipper's declaration which is required. Robyn OHSU >>> "Luck, Greg D." 11/30/2004 1:59:19 PM >>> Richard, Would you further define or point me towards a regulatory definition of "Dangerous Goods". Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 8:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Wed, 01 Dec 2004 11:16:24 -0800 From: "Robyn Vazquez" Subject: RE: [Histonet] Type of hematoxylin stain with your bluing methods ? Tell all, please. [Scanned] To: Kemlo.Rogerson@elht.nhs.uk, Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Shandon Instant Hematoxylin >>> Kemlo Rogerson 12/1/2004 8:30:30 AM >>> Harris's and Gill's. -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 01 December 2004 15:30 To: Gudrun Lang; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Type of hematoxylin stain with your bluing methods? Tell all, please. [Scanned] Dear All, It would be appreciated that you say which hematoxylin you are using when you give your bluing method. For Gudrun, are you using Mayers? At 02:59 PM 11/30/2004, you wrote: >Using lithiumcarbonat is the way we do it with staining frozen sections. >In the HE-stainer we have running tapwater. And the sections are nice. >After haemalaun in special stains we let the slides in tapwater for 3-5 min. > >Gudrun Lang Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Wed, 1 Dec 2004 14:00:00 -0600 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] Enzyme stains To: , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Novocastra has a few different antibodies for cathepsin. www.novocastra.co.uk 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: BennettW@pac.dfo-mpo.gc.ca [mailto:BennettW@pac.dfo-mpo.gc.ca] Sent: Wednesday, December 01, 2004 11:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Enzyme stains Hello Histonetters, Does anyone know if there is anything that will stain an enzyme called cathepsin. This is an enzyme that is produced by a myxobilatus sp. parasite (Kudoa thyristes) found in fish. The parasite produces this protealytic enzyme. My preliminary searches haven't revealed too much more than that. Any help would be great. Cheers Bill Bennett Histologist Fisheries and Oceans Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Wed, 01 Dec 2004 14:21:16 -0600 From: "Nita Searcy" Subject: [Histonet] Milestone Microwave Processor To: Message-ID: Content-Type: text/plain; charset="us-ascii" After 18 months in service, we are experiencing paraffin being drawn into the vacuum tube - anyone else having this problem? Can anyone provide help ? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:4-2438 ORG:;Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD ------------------------------ Message: 9 Date: Wed, 1 Dec 2004 14:27:40 -0600 From: "Bauer, Karen" Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? To: "'Stacy McLaughlin'" , "'Bauer, Karen'" , "'Richard Cartun'" , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Stacy, If you go to the CAP TODAY, Oct. 2004 issue on page 101, there is a Q & A about shipping of materials. It is also on the CAP General Checklist # GEN.40522. CAP will be changing their training requirements to every two years instead of the "annual training" that they have on the checklist now. Hope this helps, Karen -----Original Message----- From: Stacy McLaughlin [mailto:Stacy_McLaughlin@cooley-dickinson.org] Sent: Wednesday, December 01, 2004 1:06 PM To: 'Bauer, Karen'; 'Richard Cartun'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Is anyone checking that this requirement is being fulfilled? (CAP, JCAHO, OSHA, etc...) I want to make sure the right thing is being done, but I also would like to know if anyone is watching for that specifically. Thanks, Stacy McLaughlin HT (ASCP) (and Lab Safety Lady) Cooley Dickinson Hospital Northampton,MA -----Original Message----- From: Bauer, Karen [mailto:Bauer.Karen@mayo.edu] Sent: Wednesday, December 01, 2004 12:42 PM To: 'Richard Cartun'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Paraffin blocks - Dangerous goods? Our safety person in the lab has just "trained" us all on the proper packaging of specimens in the lab based on the new regulations that are out. Regulations come from the International Civil Aviation Organization (ICAO), the International Air Transport Association (IATA), the USPS, and the Department of Transportation (DOT). Risk groups are ranged from 1 to 4 with 1 being "No to Low Risk" and 4 being "High Risk". Low risk specimens are things like diagnostic serum, whole blood, urine, or tissue samples not known to contain pathogens. Packaging for these specimens need to have a primary container (a bio-hazard bag would be fine for tissue blocks, or maybe the paraffin block itself is the primary container since the tissue is embedded inside it. I'll have to check on that...), then a secondary container (which could be another bio-hazard bag), and then an outer package that is sturdy. This could be a cardboard box or one of those insulated manila envelopes used for fragile items. We do not need to really worry about the outer package, since the couriers that pick up our specimens from the lab have coolers or bio-hazard containers that they put the specimens into. This is considered the "outer package". Our safety person then had everyone who went through the presentation take a short quiz (for competency) and gave us a "Certificate" of being trained. This training will need to be done every 2 years. Everyone in the lab is being trained, since at some point we will all need to know how to ship something out whether it will be Diagnostic or Infectious Specimens (Risk 1 to Risk 4). Karen Bauer HT(ASCP Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 30, 2004 10:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin blocks - Dangerous goods? I have just been told that paraffin tissue blocks are considered "Dangerous Goods" and can only be packaged and shipped by individuals who have received special training. Would anyone like to comment on this? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. 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If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail from your computer system. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From bhewlett <@t> cogeco.ca Fri Dec 3 09:59:39 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Overfixed tissues for IHC References: Message-ID: <007501c4d951$18a45de0$6400a8c0@mainbox> Jackie, In general, there is NO such thing as 'overfixation' in formaldehyde. The statement should qualify as an urban myth! That said, we have tested most clinically applicable antibodies on human tissues fixed for up to 1 year in phosphate buffered formaldehyde and have found no significant diminishment of immunoreactivity. One caveat, the immuno protocol must have been optimized on control tissues fixed for a similar time to the test material. In general, this means that any 'antigen retrieval' performed prior to immunostaining usually has to have a somewhat extended application time in order to accommodate the more extensive cross-linking obtained after lengthy fixation. My advice is to go for it, bearing the above in mind! You may find the following reference of some use; Arber DA. Effect of prolonged formalin fixation on the immunohistochemical reactivity of breast markers. Appl Immunohistochem Mol Morphol. 2002; 10(2): 183-186. Regards, Bryan ----- Original Message ----- From: "Jackie M O'Connor" To: Sent: Friday, December 03, 2004 10:18 AM Subject: [Histonet] Overfixed tissues for IHC > Happy Friday - > I've been asked if tissues that have been in formalin for 5 months are > still good for IHC (I don't know what antibodies they want). > I'd appreciate your feedback on this one, anyone and everyone. > > Thanks - > Jackie O' > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From TJJ <@t> Stowers-Institute.org Fri Dec 3 10:26:30 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Request for removing digest messages when replying Message-ID: Hi folks! It would really be nice if, in addition to "When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..."" that you also remove the digest messages. Those of us who receive the digest instead of individual messages really sometimes have to figure out what's the new digest message vs. the old ones. This last digest in particular, Vol 13, Issue 7, took me a while to forge my way through it before I could make any sense of it. It had 8 "real" messages and scrolling down this is the number scheme 1, 2, 3, 4, 5, (begin copied digest of 22 messages--1, 2, 3, 4, 5, 6, 7, 8, 9) then 6 (back to original digest), 7, 8, (begin copied digest of 22 messages 1-9 again). I was looking in particular for number 6. There were 3 of them. Ack! When I reply to a particular message on the digest, I open an new mail message, address it to histonet (never NEVER hit reply), and copy the subject line from the particular digest message I'm responding to and paste it into the subject line of my new mail message. On occasion I will include the previous message/question, but many times will just type my response. This might not make any difference to regular histonet subscribers (one message at a time), but makes a world of difference to us digest subscribers. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From gcallis <@t> montana.edu Fri Dec 3 10:36:27 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Re: Mouse Paws In-Reply-To: References: Message-ID: <6.0.0.22.1.20041203085407.01b2b290@gemini.msu.montana.edu> You processing schedule is short. We process mouse paws NBF fixed for a week, decalcified with 10% formic acid using endpoint checks until paws are done - this can take up to 3 days in stronger formic acid. Your buffered formic acid is approx 4.5% formic acid (did a calculation). Our decalcifier is made from 88% stock formic acid. Decalcification, surprisingly, can take longer than you think due to the tiny bones tightly packed inside skin with very thick pads,tendons, etc. There are some hints on how to do determinations to not have to test every paw you work with in a big study. Of course, xray is the best method for endpoint determination, but not everyone has a FAXITRON. However if you have this unit, use it for endpoint as it is the most sensitive endpoint determination and you can do ALL the paws with one shot of xray on an 8 X 10 film. If you can fix the paws so they stay FLAT with toes spread out - this helps also. When the toes curl up after dissection from radius/ulna or tibias, we have more problems. We do 70%, 95% X 2, 100% x 2, xylene 1 change, Clearite 3 X 1 change, and 4 changes of paraffin ALL reagents including paraffin are done with vacuum and pressure and only ambient temperatures. ALL stations are 2 hours per station, very extended processing. We found the density of these tiny paws required longer processing. You could start with 1 1/2 hours per station, but if the problem still exists, up the times. Our paraffin temp NEVER exceeds 60C. Opaque, mushy tissue can mean several things. 1) Incomplete decalcification, 2) incomplete dehydration (mushy syndrome!) and paraffin infiltration i.e. processing or 3) both. If animal has inflammed paws and you still have problems, then you can always add time to processing schedule. We often cut open skin on the back of paws to allow better reagent penetration/echange. Take care to not damage an area you want to see; this might work on pad side also. Do this at collection, it is easier to slide fine point of scissors under unfixed skin and helps with fixation too. Use ultra fine point stainless steel cuticle scissors found in nail care aka glamour section of Walmart, Target, grocery/drugstores - made by LaCrosse and cost $7 to $9 (frequently on sale) - far cheaper than most vendors. If you use Tissue Prep 2, a harder paraffin, this is a good selection for bone work for both infiltration and embedding. We use it and find it works for soft tissues without sectioning problems. Good luck At 07:57 AM 12/3/2004, you wrote: >Hello Histonetters, > >I'm looking for some help or guidance in obtaining good sections from >mouse front paws, to include the bones of the wrist and digits, as well as >maintaining the surrounding matrix. This is an ongoing problem in our >laboratory and we have a difficult time obtaining good sections at this >level, the problem appearing to be inadequate infiltration of paraffin >into the interior of the paws, leaving the center somewhat mushy & white. > >The feet are fixed in 10% NBF for up to a week, decalcified in formic >acid/sodium citrate for 3 to 5 days, processed on a Shandon Pathcentre for >45 minutes/reagent, 4 changes of paraffin, 45 minutes each, vacum & >pressure on the waxes only. > >Any suggestions regarding fixation, decalcification, or processing >enhancements to improve the overall picture would be extremely helpful. > >Thanks! > >Tom Crowell >BiogenIdec >Cambridge, MA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Sherif <@t> pathwaydx.com Fri Dec 3 11:12:22 2004 From: Sherif <@t> pathwaydx.com (Sherif Girees) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] RE: Histonet Digest, Vol 13, Issue 6 Message-ID: <54FBFBCA3A8E2147884BB5ECEAB7114C609DD5@pathwaysrv.PathwayCorp.local> To Darren Hess, I used PAP pens from Vector Laboratories for years and they work great, no reaction what so ever. Sherif Girees,B.S.HT(ASCP)QIHC Manager of Molecular Pathology. Pathway Diagnostics Inc. 11215 Knott Avenue. Cypess, Ca 90630. Tel. 714-892-1201. Fax 714-892-1211. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Friday, December 03, 2004 8:25 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 13, Issue 6 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: New JCAHO PI Standard (Horn, Hazel V) 2. immunohistochemical marker for osteoblasts (Elizabeth Chlipala) 3. [BULK] - RE: [Histonet] static electricity (Fred Underwood) 4. X-gal and pap pens? (hessd101@comcast.net) 5. RE: fixation for RE: Masson's Trichrome on frozen heart sections (GUTIERREZ, JUAN) 6. RE: static electricity/humidity problems (rgrow@bmnet.com) 7. Re: X-gal and pap pens? (Gayle Callis) 8. RE: Destaining DifQuik (Tony Henwood) 9. C4D Vendor (WWmn916@aol.com) 10. RE: RE: static electricity/humidity problems (bettina.hutz@orionpharma.com) 11. RE: Destaining DifQuik[Scanned] (Kemlo Rogerson) 12. RE: Destaining DifQuik[Scanned] (Kemlo Rogerson) 13. RE: M.tuberculosis in formalin-fixed tissue[Scanned] (Kemlo Rogerson) 14. cryosection fetal tissue (Arvind Pundir) 15. Re: Re: Ammonia water pH and other bluing solutions[Scanned] (lpwenk@sbcglobal.net) 16. RE: Re: Ammonia water pH and other bluing solutions[Sc anned] (Kemlo Rogerson) 17. RE: RE: static electricity/humidity problems (Weems, Joyce) 18. RE: C4D Vendor (GUTIERREZ, JUAN) 19. RE: C4D Vendor (Sebree Linda A.) 20. RE: RE: static electricity/humidity problems (Philip Oshel) 21. RE: RE: static electricity/humidity problems (Weems, Joyce) ---------------------------------------------------------------------- Message: 1 Date: Thu, 2 Dec 2004 12:08:14 -0600 From: "Horn, Hazel V" Subject: RE: [Histonet] New JCAHO PI Standard To: "Nita Searcy" , histonet@lists.utsouthwestern.edu Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B9BD@EMAIL.archildrens.org> Content-Type: text/plain; charset=us-ascii Our pathologists do this. All of our cases are assigned a code and a report is pulled up every month using the codes for review by the surgical affair committee. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Nita Searcy Sent: Thursday, December 02, 2004 7:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New JCAHO PI Standard There is a new standard that states: "An analysis is performed for all major discrepancies between preoperative and postoperative (including pathologic) diagnoses." How are institutions complying ? In my past lives there were "Tissue Committees" that reviewed "normal" tissues but this standard , in my opinion, would not be able to be handled in the pathology lab. We consider ourselves lucky if we get any preoperative diagnosis on our requisitions received from surgery. Perhaps some kind of chart review on all surgeries? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== ------------------------------ Message: 2 Date: Thu, 2 Dec 2004 11:14:47 -0700 From: "Elizabeth Chlipala" Subject: [Histonet] immunohistochemical marker for osteoblasts To: "'Histonet'" Message-ID: <000001c4d89a$ce98a550$76d48a80@AMY> Content-Type: text/plain; charset="us-ascii" Is anyone aware of a Immunohistochemical marker that is specific for osteoblasts? I have seen references on BAP (bone alkaline phosphatase) and some other clones such as 2D3 and 2H10, but I have been unable to find a source. Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 ------------------------------ Message: 3 Date: Thu, 02 Dec 2004 13:35:22 -0500 From: "Fred Underwood" Subject: [BULK] - RE: [Histonet] static electricity To: , , Message-ID: Content-Type: text/plain; charset=US-ASCII Static Guard (brand name) aerosol spray works well. Spray a small amount on your knife clamp and other "sticky" microtome surfaces. >>> 12/02/04 12:26AM >>> Hello, I have the same problem during winter time - and in summer time it is the opposite: it is "common accepted" that in July/August there is hardly any staining possible, because the huminidy goes immediately to the alcohol and then the xylene is milky and mounting not proper. When I have been working in Germany for 10 years I never has this effect, but since I work in Finland I am fighting with it every summer. So, if anyone has a solution to the humidity problem, please share it with me:) Tina -----Original Message----- From: Pereira, Laurie [mailto:Laurie.Pereira@sdcounty.ca.gov] Sent: Thursday, December 02, 2004 1:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] static electricity Hey all, I'm new at Histology and am having a problem with static electricity. During the winter, everything tends to stick to me. I've heard that a humidifier works pretty well but our laboratory is pretty large. Will a humidifier work in such a large space? Does anyone have any hints on how to deal with the wonders of static electricity? Any information would be extremely helpful! Thanks Frustrated _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Thu, 02 Dec 2004 18:55:55 +0000 From: hessd101@comcast.net Subject: [Histonet] X-gal and pap pens? To: histonet@lists.utsouthwestern.edu Message-ID: <120220041855.7738.41AF653B000591D600001E3A2200761438CECFCE0B9C9C0A08@comcast.net> Content-Type: text/plain I've searched far and wide for a solution to this problem, and I hope the experts can finally help me. Our lab performs x-gal staining on tissue sections. Prior to 6 months ago, we used Kiyota's Super Pap Pen (which put down an excellent, CLEAR barrier). Our last pen ran (mostly) dry, and we tried to order new pens from Kiyota. Apparently, their clear pens were discontinued because of toxicity, and now the only pen available from them is the Super HT Pap Pen. The new pens put down a flimsy, blue-green colored barrier. The trouble is this - when X-gal staining, some component of the stain reacts with the dye in the pap pen producing a grainy red precipitate which discolors the slide, tissue, etc. In addition, an oily reidue floats up on the solution and can stick to tissue while removing it. In a nutshell, the new pens are awful. Does anyone know of a clear pen that is still sold (EMS, zymed all seem to have colored pens), or have experience with a colored pap pen that doesn't react with xgal staining solution? Thanks Darren Hess MD/PhD student, Dept. of Neuroscience University of Pennsylvania School of Medicine ------------------------------ Message: 5 Date: Thu, 2 Dec 2004 14:05:54 -0600 From: "GUTIERREZ, JUAN" Subject: [Histonet] RE: fixation for RE: Masson's Trichrome on frozen heart sections To: "Gayle Callis" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" After sectioning we put the slide in a coplin jar with alcoholic formalin for 5 minutes, and proceed with the stain. 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: Gayle Callis [mailto:gcallis@montana.edu] Sent: Thursday, December 02, 2004 11:20 AM To: GUTIERREZ, JUAN; Histonet@lists.utsouthwestern.edu Subject: fixation for RE: Masson's Trichrome on frozen heart sections Juan, How do you fix fresh tissue frozen sections? Bouins or NBF? and fixation times? If we were to do this stain, our tissues are NEVER prefixed before snap freezing. A one step staining procedure makes a lot of sense when handling more fragile frozen sections. I know one substitute light green with aniline blue, or vice versa - what ever one likes best. This was discussed just a few months ago on Histonet. At 09:48 AM 12/2/2004, you wrote: >We use Gomori's trichrome on frozen sections. It's a one step procedure >that gives you a faster answer. Are you able to try alternatives to MTS? >Good luck. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 6 Date: Thu, 2 Dec 2004 16:48:43 -0500 From: rgrow@bmnet.com Subject: [Histonet] RE: static electricity/humidity problems To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Tina, There have been several good responses regarding static. Try them. Use what works best in your lab. Humidity is a big problem in the southern United States. Air conditioning systems and de-humidifiers can't handle the load in old drafty buildings. We have solved this problem by adding silica gel crystals to our last absolute alcohol and first xylene in the dehydrating and clearing stations. This step has been successfully used here for many years. It also extends the life of the reagents 2x. We order it from: Sigma Aldrich Company, Catalog #: 21,444.2 Silica Gel 2.5 Kg. A word of caution: Do dispense and discard carefully as it is a carcinogen. Good luck. 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: bettina.hutz@orionpharma.com [mailto:bettina.hutz@orionpharma.com] Sent: 02 December 2004 05:26 To: Laurie.Pereira@sdcounty.ca.gov; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] static electricity[Scanned] Hello, I have the same problem during winter time - and in summer time it is the opposite: it is "common accepted" that in July/August there is hardly any staining possible, because the huminidy goes immediately to the alcohol and then the xylene is milky and mounting not proper. When I have been working in Germany for 10 years I never has this effect, but since I work in Finland I am fighting with it every summer. So, if anyone has a solution to the humidity problem, please share it with me:) Tina ------------------------------ Message: 7 Date: Thu, 02 Dec 2004 15:26:32 -0700 From: Gayle Callis Subject: Re: [Histonet] X-gal and pap pens? To: hessd101@comcast.net, Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041202150331.01b51298@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Darren, Try the Vector ImmEdge pens, they are very clean, dry nicely, and have long since replaced Kiyota's pens in our lab. Vector sells their pens in package of two, at a price of one pen from other vendors. One thing to do when using the Immedge pen - shake the begolly's out of them to disperse the active marking goo. Their technical rep recommended this be done. I actually vortex mine to insure goo redistribution. It is interesting that Kiyota's Super Pap Pens were declared toxic. I had wondered about this ( after noticing some very strong solvent odors) and asked them about it, but they said they were acceptable, but that fact changed somewhere along the line. DAKO has a clear pen, but be careful when dispensing - if you press toooooo hard, you have a Niagara Falls of goo all over everything. I macerated ( with a pliers!) their hard tip to make it a tidge fuzzier to capture the goo rush and also pulled a tip from a favorite pen and did a trade of hard tip with softer tip. We also let DAKO pen marks dry longer, but I have that luxury of time and with frozen sections before staining and/or fixation. Hopefully DAKO corrected this over-dispensing problem as the goo is good and it is clear. If all else fails, you can buy hydrophobic barrier slides with lines in a circle, squares or rectangles that work quite well. Just mount the section inside lines and do the staining. Erie has a huge selection of these, and we have even had custom hydrophobic barrier slides made for us to eliminate pap pens. Lovely thing is the barrier stays put unlike the pen marking, and the lines we liked were the one that are flatter like the Erie control slides so mounting a coverslip is more flush with slide surface. You can always talk to Erie about their selection. At 11:55 AM 12/2/2004, you wrote: >I've searched far and wide for a solution to this problem, and I hope the >experts can finally help me. Our lab performs x-gal staining on tissue >sections. Prior to 6 months ago, we used Kiyota's Super Pap Pen (which put >down an excellent, CLEAR barrier). Our last pen ran (mostly) dry, and we >tried to order new pens from Kiyota. Apparently, their clear pens were >discontinued because of toxicity, and now the only pen available from them >is the Super HT Pap Pen. The new pens put down a flimsy, blue-green >colored barrier. The trouble is this - when X-gal staining, some component >of the stain reacts with the dye in the pap pen producing a grainy red >precipitate which discolors the slide, tissue, etc. In addition, an oily >reidue floats up on the solution and can stick to tissue while removing >it. In a nutshell, the new pens are awful. Does anyone know of a clear pen >that is still sold (EMS, zymed all seem to have colored pens), or have >experience with a colored pap pen that doesn'! > t react with xgal staining solution? Thanks > >Darren Hess >MD/PhD student, Dept. of Neuroscience >University of Pennsylvania School of Medicine >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 8 Date: Fri, 3 Dec 2004 09:46:08 +1100 From: Tony Henwood Subject: RE: [Histonet] Destaining DifQuik To: "'Mary North'" , histonet@lists.utsouthwestern.edu Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E12B@simba.kids> Content-Type: text/plain To destain Romanowsky type stains, place in 1% acetic acid in 90% ethanol. If the smears have been air-dried prior to DifQuik staining, then PAP staining will be very dissapointing since the cells will be affected by air-drying. There is no technique known to reverse this. Regards Tony Henwood JP, 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: Mary North [mailto:northma@ohsu.edu] Sent: Friday, 3 December 2004 3:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Destaining DifQuik Has anyone tried to destain DifQuik-stained cytology preps so that it can be restained with PAP? Mary North, HT(ASCP), HTL OHSU Portland, OR _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** ------------------------------ Message: 9 Date: Thu, 2 Dec 2004 23:05:01 EST From: WWmn916@aol.com Subject: [Histonet] C4D Vendor To: histonet@pathology.swmed.edu Message-ID: <88.1affec28.2ee13fed@aol.com> Content-Type: text/plain; charset="US-ASCII" Greetings everyone, Has anyone heard if there is a C4D vendor? Is there anything on the horizon for availability? Next question.....it may be simple, but does HIPAA rules say patient names can not be on slides? Thanks, Deb King, HT(ASCP) Sacramento, CA ------------------------------ Message: 10 Date: Fri, 3 Dec 2004 09:51:35 +0200 From: bettina.hutz@orionpharma.com Subject: RE: [Histonet] RE: static electricity/humidity problems To: rgrow@bmnet.com, histonet@lists.utsouthwestern.edu Message-ID: <243E1A79AF5D89438C70513DBADAA4F0019E3FFD@sfies-exchange1.orionnet.org> Content-Type: text/plain; charset="iso-8859-1" Hello Renee, this sounds good, I WILL try it:) Thanks a lot, "kiitos" as we say in finnish:) Tina -----Original Message----- From: rgrow@bmnet.com [mailto:rgrow@bmnet.com] Sent: Thursday, December 02, 2004 11:49 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: static electricity/humidity problems Tina, There have been several good responses regarding static. Try them. Use what works best in your lab. Humidity is a big problem in the southern United States. Air conditioning systems and de-humidifiers can't handle the load in old drafty buildings. We have solved this problem by adding silica gel crystals to our last absolute alcohol and first xylene in the dehydrating and clearing stations. This step has been successfully used here for many years. It also extends the life of the reagents 2x. We order it from: Sigma Aldrich Company, Catalog #: 21,444.2 Silica Gel 2.5 Kg. A word of caution: Do dispense and discard carefully as it is a carcinogen. Good luck. 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: bettina.hutz@orionpharma.com [mailto:bettina.hutz@orionpharma.com] Sent: 02 December 2004 05:26 To: Laurie.Pereira@sdcounty.ca.gov; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] static electricity[Scanned] Hello, I have the same problem during winter time - and in summer time it is the opposite: it is "common accepted" that in July/August there is hardly any staining possible, because the huminidy goes immediately to the alcohol and then the xylene is milky and mounting not proper. When I have been working in Germany for 10 years I never has this effect, but since I work in Finland I am fighting with it every summer. So, if anyone has a solution to the humidity problem, please share it with me:) Tina _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Fri, 3 Dec 2004 08:13:48 -0000 From: Kemlo Rogerson Subject: RE: [Histonet] Destaining DifQuik[Scanned] To: histonet@lists.utsouthwestern.edu Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B56F@bhrv-nt-11.bhrv.nwest.nhs.uk> Content-Type: text/plain Diff-Quick is usually carried out on air-dried preps and PAP on alcohol fixed. You can destain Diff-quick in 1% acid alcohol or just by leaving in alcohol, but the PAP will be hopeless as it was air-dried. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Mary North [mailto:northma@ohsu.edu] Sent: 02 December 2004 16:40 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Destaining DifQuik[Scanned] Has anyone tried to destain DifQuik-stained cytology preps so that it can be restained with PAP? Mary North, HT(ASCP), HTL OHSU Portland, OR _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Fri, 3 Dec 2004 08:22:12 -0000 From: Kemlo Rogerson Subject: RE: [Histonet] Destaining DifQuik[Scanned] To: histonet@lists.utsouthwestern.edu Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B571@bhrv-nt-11.bhrv.nwest.nhs.uk> Content-Type: text/plain Oops sorry, you already said it! That's the trouble being in the UK, life has passed you by as you sleep. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Tony Henwood [mailto:AnthonyH@chw.edu.au] Sent: 02 December 2004 22:46 To: 'Mary North'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Destaining DifQuik[Scanned] To destain Romanowsky type stains, place in 1% acetic acid in 90% ethanol. If the smears have been air-dried prior to DifQuik staining, then PAP staining will be very dissapointing since the cells will be affected by air-drying. There is no technique known to reverse this. Regards Tony Henwood JP, 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: Mary North [mailto:northma@ohsu.edu] Sent: Friday, 3 December 2004 3:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Destaining DifQuik Has anyone tried to destain DifQuik-stained cytology preps so that it can be restained with PAP? Mary North, HT(ASCP), HTL OHSU Portland, OR _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Fri, 3 Dec 2004 08:31:14 -0000 From: Kemlo Rogerson Subject: RE: [Histonet] M.tuberculosis in formalin-fixed tissue[Scanned] To: histonet@lists.utsouthwestern.edu Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B572@bhrv-nt-11.bhrv.nwest.nhs.uk> Content-Type: text/plain No-one replied to this yet? No? Ok. TB lives (hibernates) in bone, that's why, in the old days people were sent to Sanatoriums. They recovered from their TB, by good diet etc., but when they were released and their diet/ health got worse, the bone de-ossified and the TB was released; they then suffered a relapse. I assume that can happen in lung etc., as the TB may be encapsulated in areas of bony spiculues (can't spell that) it is shielded from the fixative. I assume staining, cutting, etc., can release the bacteria from its bony coffin to make the sections infective. I would assume the risk is low, but there all the same. There is an 'explosion' of TB in London, I gather from the News today. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Therersa Stegall [mailto:STEGTM@samcstl.org] Sent: 02 December 2004 17:07 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] M.tuberculosis in formalin-fixed tissue[Scanned] Say y'all. I have a copy of CAP Today in front of me, last months'. There's an article on pg118 that says somewhere around 10% of cultured lung tissue, fixed in formalin, grew M.tuberculosis. We all assume the risk when cutting frozens, and the subsequent decontamination procedures; has anyone really thought about this? Have you heard anything else about such? Does anyone out there assume that this fixed tissue is still viable (we assume that it is a mere capsular ghost of itself). I'd be interested to hear how you deal with this type of tissue; precautions or not...... Terre _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Fri, 3 Dec 2004 14:34:53 +0530 From: "Arvind Pundir" Subject: [Histonet] cryosection fetal tissue To: Message-ID: <7DE0291A47C4BC4D9CDEF65D5017CEF0A704@mail.nbrc.res.in> Content-Type: text/plain; charset="utf-8" can any one pls tell how to embed and cut human fetal tissue for immuno as it is too soft to cut thanks in advance Arvind Singh Pundir National Brain Research Centre Near NSG Campus Manesar, Haryana - 122 050, India arvind@nbrc.res.in Tel.: 91-124 ?EUR" 233 8922-26 Fax: 91-124 - 233 89 10 / 91-124 - 233 89 28 aNational Brain Research Centre Manesar, Gurgaon DiNational Brain Research Centre Manesar, Gurgaon Dist. Haryana-122 050, India st. Haryana-122 050, India rch Centre Manesar, Gurgaon Dist. Haryana-122 050, India National Brain Research Centre Manesar, Gurgaon Dist. Haryana-122 050, India National Brain Research Centre Manesar, Gurgaon Dist. Haryana-122 050, India National Brain Research Centre Manesar, Gurgaon Dist. Haryana-122 050, India National Brain Research Centre Manesar, Gurgaon Dist. Haryana-122 050, India ------------------------------ Message: 15 Date: Fri, 3 Dec 2004 05:37:58 -0500 From: Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions[Scanned] To: "Kemlo Rogerson" , "'Jackie M O'Connor'" , "Johnson, Teri" Cc: Histonet Message-ID: <004d01c4d924$29272da0$2329d445@domainnotset.invalid> Content-Type: text/plain; charset="iso-8859-1" Let's see if I can explain pH, hematoxylin, red vs. blue, and bluing, and not get myself into a murky mess of chemistry. Hematoxylin solution is actually an aluminum salt and oxidized hematoxylin, collectively known as aluminum-hematein (Al-hematein). In order to stain just the DNA and RNA, the pH of the Al-hematein solution needs to be between 2.2 to 2.9 (most usually around 2.4-2.5). At this lower (acidic) pH, there are a lot of hydrogen ions (H+) in the Al-Hematein solution. These excess H+ bind to the Al-hematein, causing a shift to the red. When the stained tissue is placed in a solution more alkaline than the 2.2-2.9 stain, there are fewer H+ in the staining solution. This removes the H+ from the Al-Hematein. This causes a color shift in the dye to the blue range (i.e., "bluing the slides"). Therefore, bluing agents must have a pH higher than the mid-2.5. The higher the pH (more alkaline), the faster the H+ are removed from the Al-Hematein, and the faster the "bluing" will take place. Bluing at about pH 5.0 can take up to about 10 minutes to "blue" totally (all red nuclei to some red/some blue (= purple looking tissue) to all blue nuclei). At pH 10, it will take 30 seconds. Tap water is more alkaline than the Al-hematein, so it can be used to blue slides. If the tap water's pH is around 7, this should take about 5 minutes. Some lab's water is more alkaline than other labs, so will take less time. The pH of tap water can change depending upon what chemicals the water treatment facility is using that day to purify the water, so using just tap water to blue may have some variability of time. Most labs rely on an alkaline solution (dilute ammonia, dilute lithium carbonate, Scott's tap water) to speed up the bluing processing and to not rely upon the variability of water pH. Hope this helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Kemlo Rogerson" To: "'Jackie M O'Connor'" ; "Johnson, Teri" Cc: "Histonet" ; Sent: Wednesday, December 01, 2004 3:18 AM Subject: RE: [Histonet] Re: Ammonia water pH and other bluing solutions[Scanned] > I think you are wrong, so I'll correct you, as you requested. > > I thought the 'blue' state of haematin was, like litmus, the effect an > alkaline condition has on the siderphilic dye. Red, like litmus, denoted > acidic conditions. Damned if I know what colour (with the 'u') haematin goes > when neutral (bluey/ red?) > > Running tap water takes longer, I concede, but good things are worth waiting > for and it's easier to make up; we rush too much I find. TIP: Never use > London water, been through too many kidneys! > > -----Original Message----- > From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] > Sent: 30 November 2004 19:09 > To: Johnson, Teri > Cc: Histonet; histonet-bounces@lists.utsouthwestern.edu > Subject: Re: [Histonet] Re: Ammonia water pH and other bluing > solutions[Scanned] > > Anyone remember using saturated lithium carbonate? I could probably use a > little lithium right now . . . . . > > Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just > bringing the slides BACK to a neutral pH after treating them with acid > which makes them purple-ish? I like the ammonia because it 'seems' to > make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to > make them "blah".(Perception is in the eye of the beholder) Running tap > water will make the same miracle happen, but it takes longer. > > > > > > > "Johnson, Teri" > Sent by: histonet-bounces@lists.utsouthwestern.edu > 11/30/2004 12:51 PM > > > To: "Histonet" > cc: > Subject: [Histonet] Re: Ammonia water pH and other bluing > solutions > > > Way back in the day, Brigati was using PBS on his immunostainer to blue > after the hematoxylin counterstain. I have since used PBS to blue my > slides instead of ammonium hydroxide/water. It's cheap, and very easy > on the sections, and I have plenty of it on hand. > > > Teri Johnson > Managing Director Histology Facility > Stowers Institute for Medical Research > 1000 E. 50th St. > Kansas City, Missouri 64110 > tjj@stowers-institute.org > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Fri, 3 Dec 2004 10:47:26 -0000 From: Kemlo Rogerson Subject: RE: [Histonet] Re: Ammonia water pH and other bluing solutions[Sc anned] To: Histonet Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B573@bhrv-nt-11.bhrv.nwest.nhs.uk> Content-Type: text/plain Oh I see now, so why are nuclei blue? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] Sent: 03 December 2004 10:38 To: Kemlo Rogerson; 'Jackie M O'Connor'; Johnson, Teri Cc: Histonet Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions[Scanned] Let's see if I can explain pH, hematoxylin, red vs. blue, and bluing, and not get myself into a murky mess of chemistry. Hematoxylin solution is actually an aluminum salt and oxidized hematoxylin, collectively known as aluminum-hematein (Al-hematein). In order to stain just the DNA and RNA, the pH of the Al-hematein solution needs to be between 2.2 to 2.9 (most usually around 2.4-2.5). At this lower (acidic) pH, there are a lot of hydrogen ions (H+) in the Al-Hematein solution. These excess H+ bind to the Al-hematein, causing a shift to the red. When the stained tissue is placed in a solution more alkaline than the 2.2-2.9 stain, there are fewer H+ in the staining solution. This removes the H+ from the Al-Hematein. This causes a color shift in the dye to the blue range (i.e., "bluing the slides"). Therefore, bluing agents must have a pH higher than the mid-2.5. The higher the pH (more alkaline), the faster the H+ are removed from the Al-Hematein, and the faster the "bluing" will take place. Bluing at about pH 5.0 can take up to about 10 minutes to "blue" totally (all red nuclei to some red/some blue (= purple looking tissue) to all blue nuclei). At pH 10, it will take 30 seconds. Tap water is more alkaline than the Al-hematein, so it can be used to blue slides. If the tap water's pH is around 7, this should take about 5 minutes. Some lab's water is more alkaline than other labs, so will take less time. The pH of tap water can change depending upon what chemicals the water treatment facility is using that day to purify the water, so using just tap water to blue may have some variability of time. Most labs rely on an alkaline solution (dilute ammonia, dilute lithium carbonate, Scott's tap water) to speed up the bluing processing and to not rely upon the variability of water pH. Hope this helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Kemlo Rogerson" To: "'Jackie M O'Connor'" ; "Johnson, Teri" Cc: "Histonet" ; Sent: Wednesday, December 01, 2004 3:18 AM Subject: RE: [Histonet] Re: Ammonia water pH and other bluing solutions[Scanned] > I think you are wrong, so I'll correct you, as you requested. > > I thought the 'blue' state of haematin was, like litmus, the effect an > alkaline condition has on the siderphilic dye. Red, like litmus, denoted > acidic conditions. Damned if I know what colour (with the 'u') haematin goes > when neutral (bluey/ red?) > > Running tap water takes longer, I concede, but good things are worth waiting > for and it's easier to make up; we rush too much I find. TIP: Never use > London water, been through too many kidneys! > > -----Original Message----- > From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] > Sent: 30 November 2004 19:09 > To: Johnson, Teri > Cc: Histonet; histonet-bounces@lists.utsouthwestern.edu > Subject: Re: [Histonet] Re: Ammonia water pH and other bluing > solutions[Scanned] > > Anyone remember using saturated lithium carbonate? I could probably use a > little lithium right now . . . . . > > Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step just > bringing the slides BACK to a neutral pH after treating them with acid > which makes them purple-ish? I like the ammonia because it 'seems' to > make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to > make them "blah".(Perception is in the eye of the beholder) Running tap > water will make the same miracle happen, but it takes longer. > > > > > > > "Johnson, Teri" > Sent by: histonet-bounces@lists.utsouthwestern.edu > 11/30/2004 12:51 PM > > > To: "Histonet" > cc: > Subject: [Histonet] Re: Ammonia water pH and other bluing > solutions > > > Way back in the day, Brigati was using PBS on his immunostainer to blue > after the hematoxylin counterstain. I have since used PBS to blue my > slides instead of ammonium hydroxide/water. It's cheap, and very easy > on the sections, and I have plenty of it on hand. > > > Teri Johnson > Managing Director Histology Facility > Stowers Institute for Medical Research > 1000 E. 50th St. > Kansas City, Missouri 64110 > tjj@stowers-institute.org > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@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, 3 Dec 2004 07:50:37 -0500 From: "Weems, Joyce" Subject: RE: [Histonet] RE: static electricity/humidity problems To: , , Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507601@sjhaexc02.sjha.org> Content-Type: text/plain; charset="iso-8859-1" And Cardinal has a product called Molecular Sieve - 4494-500ny that works the same way without the carcinogen factor. (At least as far as I know!) Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of bettina.hutz@orionpharma.com Sent: Friday, December 03, 2004 2:52 AM To: rgrow@bmnet.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: static electricity/humidity problems Hello Renee, this sounds good, I WILL try it:) Thanks a lot, "kiitos" as we say in finnish:) Tina -----Original Message----- From: rgrow@bmnet.com [mailto:rgrow@bmnet.com] Sent: Thursday, December 02, 2004 11:49 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: static electricity/humidity problems Tina, There have been several good responses regarding static. Try them. Use what works best in your lab. Humidity is a big problem in the southern United States. Air conditioning systems and de-humidifiers can't handle the load in old drafty buildings. We have solved this problem by adding silica gel crystals to our last absolute alcohol and first xylene in the dehydrating and clearing stations. This step has been successfully used here for many years. It also extends the life of the reagents 2x. We order it from: Sigma Aldrich Company, Catalog #: 21,444.2 Silica Gel 2.5 Kg. A word of caution: Do dispense and discard carefully as it is a carcinogen. Good luck. 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: bettina.hutz@orionpharma.com [mailto:bettina.hutz@orionpharma.com] Sent: 02 December 2004 05:26 To: Laurie.Pereira@sdcounty.ca.gov; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] static electricity[Scanned] Hello, I have the same problem during winter time - and in summer time it is the opposite: it is "common accepted" that in July/August there is hardly any staining possible, because the huminidy goes immediately to the alcohol and then the xylene is milky and mounting not proper. When I have been working in Germany for 10 years I never has this effect, but since I work in Finland I am fighting with it every summer. So, if anyone has a solution to the humidity problem, please share it with me:) Tina _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. ------------------------------ Message: 18 Date: Fri, 3 Dec 2004 07:41:53 -0600 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] C4D Vendor To: , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Quidel Corporation 265 N. Whisman Rd. Mountain View, CA 94043 (800)524-6318 We use ours for frozen section IHC. 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: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Thursday, December 02, 2004 10:05 PM To: histonet@pathology.swmed.edu Subject: [Histonet] C4D Vendor Greetings everyone, Has anyone heard if there is a C4D vendor? Is there anything on the horizon for availability? Next question.....it may be simple, but does HIPAA rules say patient names can not be on slides? Thanks, Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Fri, 3 Dec 2004 07:59:49 -0600 From: "Sebree Linda A." Subject: RE: [Histonet] C4D Vendor To: Cc: "Histonet \(E-mail\)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Biogenesis, (603) 642-8302 Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of WWmn916@aol.com Sent: Thursday, December 02, 2004 10:05 PM To: histonet@pathology.swmed.edu Subject: [Histonet] C4D Vendor Greetings everyone, Has anyone heard if there is a C4D vendor? Is there anything on the horizon for availability? Next question.....it may be simple, but does HIPAA rules say patient names can not be on slides? Thanks, Deb King, HT(ASCP) Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 20 Date: Fri, 03 Dec 2004 08:28:22 -0600 From: Philip Oshel Subject: RE: [Histonet] RE: static electricity/humidity problems To: Histonet@Pathology.swmed.edu Message-ID: Content-Type: text/plain; format=flowed; charset=us-ascii "Cardinal"? Do you have a website for this company? I've been looking for a supplier of molecular sieve that doesn't charge outrageous prices. I assume the numbers are the catalog number for the sieve -- is it 3 - 4 Angstrom? The sieve pores must be that size to trap water molecules. Thanks. Phil >And Cardinal has a product called Molecular Sieve - 4494-500ny that >works the same way without the carcinogen factor. (At least as far >as I know!) >Joyce > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >bettina.hutz@orionpharma.com >Sent: Friday, December 03, 2004 2:52 AM >To: rgrow@bmnet.com; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: static electricity/humidity problems > > >Hello Renee, > >this sounds good, I WILL try it:) >Thanks a lot, "kiitos" as we say in finnish:) >Tina > >-----Original Message----- >From: rgrow@bmnet.com [mailto:rgrow@bmnet.com] >Sent: Thursday, December 02, 2004 11:49 PM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] RE: static electricity/humidity problems > > >Tina, > >There have been several good responses regarding static. Try them. Use >what works best in your lab. > >Humidity is a big problem in the southern United States. Air conditioning >systems and de-humidifiers can't handle the load in old drafty buildings. We >have solved this problem by adding silica gel crystals to our last absolute >alcohol and first xylene in the dehydrating and clearing stations. This step >has been successfully used here for many years. It also extends the life >of the reagents 2x. We order it from: Sigma Aldrich Company, Catalog #: >21,444.2 Silica Gel 2.5 Kg. A word of caution: Do dispense and discard >carefully as it is a carcinogen. > >Good luck. > >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: bettina.hutz@orionpharma.com [mailto:bettina.hutz@orionpharma.com] >Sent: 02 December 2004 05:26 >To: Laurie.Pereira@sdcounty.ca.gov; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] static electricity[Scanned] > >Hello, > >I have the same problem during winter time - and in summer time it is the >opposite: it is "common accepted" that in July/August there is hardly any >staining possible, because the huminidy goes immediately to the alcohol and >then the xylene is milky and mounting not proper. When I have been working >in Germany for 10 years I never has this effect, but since I work in Finland >I am fighting with it every summer. So, if anyone has a solution to the >humidity problem, please share it with me:) > >Tina > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >Confidentiality Notice ** The information contained in this message >may be privileged and is confidential information intended for the >use of the addressee listed above. If you are neither the intended >recipient nor the employee or agent responsible for delivering this >message to the intended recipient, you are hereby notified that any >disclosure, copying, distribution or the taking of any action in >reliance on the contents of this information is strictly prohibited. >If you have received this communication in error, please notify us >immediately by replying to the message and deleting it from your >computer. >Thank you. Saint Josephs Health System, Inc. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) ------------------------------ Message: 21 Date: Fri, 3 Dec 2004 09:34:23 -0500 From: "Weems, Joyce" Subject: RE: [Histonet] RE: static electricity/humidity problems To: "Philip Oshel" , Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507607@sjhaexc02.sjha.org> Content-Type: text/plain; charset="iso-8859-1" Cardinal.com Yes, 4 Angstrom Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Philip Oshel Sent: Friday, December 03, 2004 9:28 AM To: Histonet@Pathology.swmed.edu Subject: RE: [Histonet] RE: static electricity/humidity problems "Cardinal"? Do you have a website for this company? I've been looking for a supplier of molecular sieve that doesn't charge outrageous prices. I assume the numbers are the catalog number for the sieve -- is it 3 - 4 Angstrom? The sieve pores must be that size to trap water molecules. Thanks. Phil >And Cardinal has a product called Molecular Sieve - 4494-500ny that >works the same way without the carcinogen factor. (At least as far >as I know!) >Joyce > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >bettina.hutz@orionpharma.com >Sent: Friday, December 03, 2004 2:52 AM >To: rgrow@bmnet.com; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] RE: static electricity/humidity problems > > >Hello Renee, > >this sounds good, I WILL try it:) >Thanks a lot, "kiitos" as we say in finnish:) >Tina > >-----Original Message----- >From: rgrow@bmnet.com [mailto:rgrow@bmnet.com] >Sent: Thursday, December 02, 2004 11:49 PM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] RE: static electricity/humidity problems > > >Tina, > >There have been several good responses regarding static. Try them. Use >what works best in your lab. > >Humidity is a big problem in the southern United States. Air conditioning >systems and de-humidifiers can't handle the load in old drafty buildings. We >have solved this problem by adding silica gel crystals to our last absolute >alcohol and first xylene in the dehydrating and clearing stations. This step >has been successfully used here for many years. It also extends the life >of the reagents 2x. We order it from: Sigma Aldrich Company, Catalog #: >21,444.2 Silica Gel 2.5 Kg. A word of caution: Do dispense and discard >carefully as it is a carcinogen. > >Good luck. > >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: bettina.hutz@orionpharma.com [mailto:bettina.hutz@orionpharma.com] >Sent: 02 December 2004 05:26 >To: Laurie.Pereira@sdcounty.ca.gov; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] static electricity[Scanned] > >Hello, > >I have the same problem during winter time - and in summer time it is the >opposite: it is "common accepted" that in July/August there is hardly any >staining possible, because the huminidy goes immediately to the alcohol and >then the xylene is milky and mounting not proper. When I have been working >in Germany for 10 years I never has this effect, but since I work in Finland >I am fighting with it every summer. So, if anyone has a solution to the >humidity problem, please share it with me:) > >Tina > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >Confidentiality Notice ** The information contained in this message >may be privileged and is confidential information intended for the >use of the addressee listed above. If you are neither the intended >recipient nor the employee or agent responsible for delivering this >message to the intended recipient, you are hereby notified that any >disclosure, copying, distribution or the taking of any action in >reliance on the contents of this information is strictly prohibited. >If you have received this communication in error, please notify us >immediately by replying to the message and deleting it from your >computer. >Thank you. Saint Josephs Health System, Inc. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 13, Issue 6 *************************************** From amarusk1 <@t> FAIRVIEW.ORG Fri Dec 3 11:30:24 2004 From: amarusk1 <@t> FAIRVIEW.ORG (ANN MARUSKA) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] pax-5 Message-ID: Hi histonetters, Is anyone using Pax5 on B5 specimens (marrow or tissue)? I am having a devil of a time to get decent staining.......it's good on AZF and formalin. Any tips would be greatly appreciated. Have a nice weekend! Ann Ann Maruska Fairview-University Medical Center Mpls. MN 55454 amarusk1@fairview.org 612-273-9119 The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including “protected health information.” If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:ANN MARUSKA N:MARUSKA;ANN TEL;WORK:612-273-9119 ORG:;LAB EMAIL;WORK;PREF;NGW:AMARUSK1@FAIRVIEW.ORG END:VCARD From histophilhuff <@t> yahoo.com Fri Dec 3 12:56:11 2004 From: histophilhuff <@t> yahoo.com (Phillip Huff) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Mouse Paws and Gayle's Knowledge in a Book In-Reply-To: <6.0.0.22.1.20041203085407.01b2b290@gemini.msu.montana.edu> Message-ID: <20041203185612.92300.qmail@web54708.mail.yahoo.com> How many people think that Gayle should write a book describing her vast (correction - extremely vast) knowledge of histochemistry and tissue sample prep? Phil --------------------------------- Do you Yahoo!? Send holiday email and support a worthy cause. Do good. From akbitting <@t> geisinger.edu Fri Dec 3 12:57:20 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] RUOs Message-ID: Does anyone have a difinitive answer as to whether you can bill for IHC studies done with RUO primary antibodies in a clinical settiing? Thanks everyone for all your help in the past. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From dcrippen <@t> buckinstitute.org Fri Dec 3 13:04:40 2004 From: dcrippen <@t> buckinstitute.org (Danielle Crippen) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Mouse Paws and Gayle's Knowledge in a Book Message-ID: <4AA34A707932424EBE2D973764D226A901D92FDE@inverness.buckcenter.org> Yes--please do Gayle!! danielle -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Phillip Huff Sent: Fri 12/3/2004 10:56 AM To: Histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Mouse Paws and Gayle's Knowledge in a Book How many people think that Gayle should write a book describing her vast (correction - extremely vast) knowledge of histochemistry and tissue sample prep? Phil --------------------------------- Do you Yahoo!? Send holiday email and support a worthy cause. Do good. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ashelynne2 <@t> yahoo.com Fri Dec 3 13:10:45 2004 From: ashelynne2 <@t> yahoo.com (linda cobb) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] discontinue subscription Message-ID: <20041203191046.3122.qmail@web51707.mail.yahoo.com> __________________________________ Do you Yahoo!? The all-new My Yahoo! - What will yours do? http://my.yahoo.com From ashelynne2 <@t> yahoo.com Fri Dec 3 13:10:45 2004 From: ashelynne2 <@t> yahoo.com (linda cobb) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] discontinue subscription Message-ID: <20041203191045.96534.qmail@web51710.mail.yahoo.com> __________________________________ Do you Yahoo!? Dress up your holiday email, Hollywood style. Learn more. http://celebrity.mail.yahoo.com From ashelynne2 <@t> yahoo.com Fri Dec 3 13:11:14 2004 From: ashelynne2 <@t> yahoo.com (linda cobb) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] discontinue subscription Message-ID: <20041203191114.96697.qmail@web51710.mail.yahoo.com> please discontinue my subscription __________________________________ Do you Yahoo!? Send holiday email and support a worthy cause. Do good. http://celebrity.mail.yahoo.com From tpmorken <@t> labvision.com Fri Dec 3 13:26:39 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] RUOs Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA257F74A@usca0082k08.labvision.apogent.com> No. In fact, you are not even supposed to report results from RUO antibodies. You can always ask the company for an ASR datasheet (assuming it's an FDA-registered company), do your own validation, and then use it to your hearts content. Tim Morken Lab Vision - Neomarkers www.labvision.com -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Friday, December 03, 2004 10:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RUOs Does anyone have a difinitive answer as to whether you can bill for IHC studies done with RUO primary antibodies in a clinical settiing? Thanks everyone for all your help in the past. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfavara <@t> niaid.nih.gov Fri Dec 3 14:13:23 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Mouse Paws and Gayle's Knowledge in a Book Message-ID: Add me to the list!!! 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: Phillip Huff [mailto:histophilhuff@yahoo.com] Sent: Friday, December 03, 2004 11:56 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Mouse Paws and Gayle's Knowledge in a Book How many people think that Gayle should write a book describing her vast (correction - extremely vast) knowledge of histochemistry and tissue sample prep? Phil --------------------------------- Do you Yahoo!? Send holiday email and support a worthy cause. Do good. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Diann.Burtis <@t> aventis.com Fri Dec 3 14:43:41 2004 From: Diann.Burtis <@t> aventis.com (Diann.Burtis@aventis.com) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Trichrome -PAS staining Message-ID: <16721C38AAB4E643BE0D89DE62BC7500080C4A@sccsmxsusr01.pharma.aventis.com> Hi! I would like to know if anyone has any experience in doing the Trichrome PAS stain for alpha, beta, and delta cells in pancreas? Also of any current methods of staining used to stain these cells together in pancreas. Diann Aventis Pharmaceuticals Route 202-206 Bridgewater, N.J. 08807 Telephone: 908-231-2115 Fax: 908-231-2217 diann.burtis@aventis.com From bhewlett <@t> cogeco.ca Fri Dec 3 14:55:50 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Trichrome -PAS staining References: <16721C38AAB4E643BE0D89DE62BC7500080C4A@sccsmxsusr01.pharma.aventis.com> Message-ID: <000801c4d97a$78c3c570$6400a8c0@mainbox> Hi Diann, Yes, what would you like to know? The current methods are almost exclusively IHC. Bryan ----- Original Message ----- From: To: Sent: Friday, December 03, 2004 3:43 PM Subject: [Histonet] Trichrome -PAS staining Hi! I would like to know if anyone has any experience in doing the Trichrome PAS stain for alpha, beta, and delta cells in pancreas? Also of any current methods of staining used to stain these cells together in pancreas. Diann Aventis Pharmaceuticals Route 202-206 Bridgewater, N.J. 08807 Telephone: 908-231-2115 Fax: 908-231-2217 diann.burtis@aventis.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Dec 3 15:15:15 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] OK you guys (writing a book) - you're ganging up on me!! Message-ID: <6.0.0.22.1.20041203140400.01b2abe8@gemini.msu.montana.edu> Dear All, OK - "youse guys"! Quit ganging up on me!! The thought has occured to me, but so far, only a thought!! Hmmm maybe a project that could finance retirement 'cause I'm not getting any younger! Happy Holidays to everyone - Gayle Callis (still chuckling!) Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From rocan <@t> mac.com Fri Dec 3 15:27:31 2004 From: rocan <@t> mac.com (Rocan) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] OK you guys (writing a book) - you're ganging up on me!! In-Reply-To: <6.0.0.22.1.20041203140400.01b2abe8@gemini.msu.montana.edu> References: <6.0.0.22.1.20041203140400.01b2abe8@gemini.msu.montana.edu> Message-ID: <23C40E80-4572-11D9-B44B-000A9589219E@mac.com> Gail: I am sure any editor can look into histonet archives and give you a nice contract! I always read your postings and I am so very impressed with your knowledge. Research hsito is a growing field and not easy. Not many things are routine in research. A nice technical book along with a good Altas of mouse tissue could make a kill. Please do think about it. Thanks ----- Dr.Rocio Sierra-Honigmann Director Engineered Wound Repair Laboratory Cedars Sinai Research Institute Davis 1091 310-423-1882 Honigmannr@cshs.org On Dec 3, 2004, at 1:15 PM, Gayle Callis wrote: > Dear All, > > OK - "youse guys"! Quit ganging up on me!! The thought has occured > to me, but so far, only a thought!! Hmmm maybe a project that could > finance retirement 'cause I'm not getting any younger! > > Happy Holidays to everyone - > > Gayle Callis (still chuckling!) > > > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nperson211 <@t> comcast.net Fri Dec 3 15:34:54 2004 From: nperson211 <@t> comcast.net (nperson211@comcast.net) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] our own Yoda? Message-ID: <120320042134.13744.41B0DBFE000364B1000035B02200734748CECECD02019C9D0A9F02@comcast.net> Teach us, she will! Chuckle away Gayle, but I sense a growing clamour for a tome of histo-wisdom. Holiday wishes will distract the group for only so long. Nancy Lemke Hermelin Brain Tumor Center Henry Ford Hospital Detroit From maria <@t> ski.org Fri Dec 3 17:16:07 2004 From: maria <@t> ski.org (Maria Mejia) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] dioxane chemical questions Message-ID: <41B0F3B7.7060302@ski.org> Hello Everyone, in the next few weeks, we plan to use very small amounts of (dioxane) to coat coverslip - 10ml -20ml at one time - 2-4 times /month. In previous experiments, the PI has tried other chemical substitutes - without success. Without the use of dioxane, a protein called conA that attaches cells to the glass will not bond properly with the coverslips and instead will wash away during testing. This question is for anyone (past or current) using dioxane, please advise, on the following procedure: I understand in order to prevent peroxidizable compounds in the dioxane, it is necessary to store the liquid material - inerting the atmosphere in storage containers with nitrogen to inhibit peroxide formation. >When using dioxane, is there another alternative to storing in nitrogen or argon atmosphere? For example, adding an inert high boiling substance like mineral oil to prevent the peroxide from concentrating to a dangerous level? Has anyone tried this? >What type of containers (metal or plastic) should be used for cryogenic process? >What type of container should be used in disposal of dioxane? Polyethylene plastic? > Can I assume that -20C is a good temperature for storage? > How long should I keep working dioxane? I read 3-12 months. > How long should I keep waste dioxane waiting for disposal pick-up? > Disposal of peroxides - should I dilute with water - what concentration of H20? > How can I detect & determine peroxides? I notice in the Laboratory Safety that they sell peroxide check paper strips. Does anyone know if these strips are reliable and accurate? > How often should we have the fume hood where dioxane will be use...checked? Every six months? Once a year (currently doing)? Nitrile (heavy weight - 8mil-12 mil) and butyl gloves will be used. I believe that's it for now - please let me know if I have forgotten anything else! Any help or information (resources, references, companies etc) anyone can provide will be greatly appreciate. regards Maria Mejia neurohistologist Smith-Kettlewell Eye Res. Inst. San Francisco, CA 94115 Email: maria@ski.org From hsvlle <@t> free.net.nz Fri Dec 3 20:28:30 2004 From: hsvlle <@t> free.net.nz (Lorraine Rolston) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Re 'static' and histonet. Message-ID: <000c01c4d9a8$f322e520$496b61cb@default> You guys rock!!I'm off to the supermarket today to get some anti-static drier 'wipes' for my microtome. Thanks!!! O O \___/ Lorraine. --- avast! Antivirus: Outbound message clean. Virus Database (VPS): 0449-0, 30/11/2004 Tested on: 4/12/2004 3:28:35 p.m. avast! - copyright (c) 1988-2004 ALWIL Software. http://www.avast.com From skippy_ette <@t> sbcglobal.net Sat Dec 4 06:29:20 2004 From: skippy_ette <@t> sbcglobal.net (Lori Cummings) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] CD 4 Vendor Message-ID: <20041204122920.57663.qmail@web80408.mail.yahoo.com> Hi! Try Novacastra- Vision BioSystems, Inc. CD4 clone 4B12 for FFPE cat # NCL-CD4-368 800-753-7264 sales and service From NLAPOINTE <@t> PARTNERS.ORG Sun Dec 5 16:18:14 2004 From: NLAPOINTE <@t> PARTNERS.ORG (Lapointe, Nathalie,Ph.D.) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Question concerning TTC staining Message-ID: <2D7FF27625F01942A2CBD6183F62694F08EAB7@PHSXMB3.partners.org> Hi, I'm a fellow student at the Brigham and Women's hospital (Boston, MA), and I need an information in order to complete a paper. TTC staining was used to measure infarct size in rats dying between 4 and 24 hours - does such staining can only be performed accurately when the heart is excised and stained immediately after death? If the hearts were allowed to sit for some variable time period in dead animals, does the measurements would not have been accurate and the data would be unreliable? Since infarct size was used as a criterion for grouping most of the other data sets, an inaccuracy introduced into this measurement may have affected nearly all other statistical comparisons presented in the paper. However from my experience, an 6-8-hour delay of TTC staining is reliable for evaluating heart infarct size in a rat. Does this probably is attributable to the slow deterioration of mitochondrial enzyme activity in nonischemic heart over this time period? The only paper that I found concerning that matter is not with the heart but with the brain (in rat): " Delayed triphenyltetrazolium chloride staining remains useful for evaluating cerebral infarct volume in a rat stroke model. Li F, Irie K, Answer MS, Fisher M. Department of Neurology, Memorial Health Care, Worcester, Massachusetts 01605, U.S.A." Does anybody have further references or info for me? Thank you very much, Dr Nathalie Lapointe From jkiernan <@t> uwo.ca Sun Dec 5 23:08:23 2004 From: jkiernan <@t> uwo.ca (jkiernan@uwo.ca) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Re: Dioxane questions (Long,with extras) In-Reply-To: <41B0F3B7.7060302@ski.org> References: <41B0F3B7.7060302@ski.org> Message-ID: <1102309703.41b3e947bf098@mail.uwo.ca> Dear Maria Mejia, Dioxane is a liquid with a 60-year histotechnical history. Your boss's potential application for attaching cells to slides is not part of the story, and may be misguided. Read on. Dioxane is a cyclic diether that boils at 101C. It is inflammable (slightly less so than ethanol) and yes, it can form explosive peroxides on prolonged exposure to air. If you buy a small bottle and don't store it when almost empty, peroxide formation should not be a risk. You can test for peroxides by adding 1 ml of freshly made 10% aqueous potassium iodide to about 10 ml of dioxane, in a small, stoppered container. Wait one hour. Formation of iodine (yellow-brown) indicates the presence of organic peroxides. The danger of explosion exists if the solvent is evaporated to dryness, because the organic peroxides do not evaporate and therefore become concentrated. With small volumes of "young" dioxane that are not being boiled to recover solutes, peroxides do not present a risk. Disposal of dioxane is the same as for other non-chlorinated flammable liquids - collection in drums for taking away and burning. If you detect peroxides in a bottle of dioxane, add some ferrous sulphate crystals (reduces peroxides to non-explosive substances) before pouring the dioxane into the waste solvents container. Having said all this, I must question your modus operandi in connection with dioxane. Dioxane will not "coat" glass (except perhaps with organic peroxides if you use old stock). As a solvent it will remove any grease that's present and it will then simply evaporate. If your "conA" is concanavalin A (the lectin from jack beans), that won't really attach cells to glass either; the lectin sticks to the surfaces of many types of cells and microorganisms by virtue of its affinity for alpha-D-mannosyl and alpha-D-glucosyl units in cell surface glycoproteins. This lectin, like many others, is bivalent, so it can make cells or bacteria clump together. Lectins are also called phytohaemagglutinins - proteins from plants that agglutinate blood cells. Some are specific for particular human blood types, notably Groups B and O[H]. Concanavalin A is a nonspecific phytohaemagglutinin; it can bind to nearly all cell surfaces and also to extracellular carbohydrates (eg collagen), glycogen, and some types of mucus. Fluorescently tagged concanavalin A was one of the first lectins to be used as a reagent in carbohydrate histochemistry, in the early 1970s. A digression follows. Back to dioxane. The chief use of dioxane in histotechnology has been as a solvent that mixes well with both water and paraffin wax. I have used it occasionally. It is toxic (drowsiness, skin irritation, renal failure, possible carcinogen etc, according to the Merck Index, which gives references). Its rather weak (quite pleasant) smell could easily go unnoticed, so this is not a liquid to use on a large scale in paraffin processing. Tetrahydrofuran can also be used as a single dehydrating+clearing solvent, but it has disadvantages similar to those of dioxane and a lower boiling point (66C), so it's more of a fire hazard than dioxane. Tertiary butyl alcohol (tert-butanol; 2-methyl-2-propanol) is an altogether safer single solvent for transition from water to wax. It's often used in processing for plant histology. The only practical drawback with t-butanol is that the 100% liquid freezes at 26C. This is not one for general use in processing machines! John Kiernan Dept of Anatomy & Cell Biology The University of Western Ontario London, Canada -------------------------------------------------------- Quoting Maria Mejia : > Hello Everyone, in the next few weeks, we plan to use very small amounts > of (dioxane) > to coat coverslip - 10ml -20ml at one time - 2-4 times /month. > > In previous experiments, the PI has tried other chemical substitutes - > without success. > Without the use of dioxane, a protein called conA that attaches cells to > the glass will not > bond properly with the coverslips and instead will wash away during > testing. > > This question is for anyone (past or current) using dioxane, please > advise, on the following > procedure: > > I understand in order to prevent peroxidizable compounds in the dioxane, it > is necessary to store the liquid material - inerting the atmosphere in > storage containers > with nitrogen to inhibit peroxide formation. > > >When using dioxane, is there another alternative to storing in > nitrogen or argon > atmosphere? > For example, adding an inert high boiling substance like mineral oil to > prevent the > peroxide from concentrating to a dangerous level? Has anyone tried this? > > >What type of containers (metal or plastic) should be used for > cryogenic process? > > >What type of container should be used in disposal of dioxane? > Polyethylene > plastic? > > > Can I assume that -20C is a good temperature for storage? > > > How long should I keep working dioxane? I read 3-12 months. > > > How long should I keep waste dioxane waiting for disposal pick-up? > > > Disposal of peroxides - should I dilute with water - what > concentration of H20? > > > How can I detect & determine peroxides? I notice in the Laboratory > Safety that they > sell peroxide check paper strips. Does anyone know if these strips > are reliable and > accurate? > > > How often should we have the fume hood where dioxane will be > use...checked? > Every six months? Once a year (currently doing)? > > Nitrile (heavy weight - 8mil-12 mil) and butyl gloves will be used. > > I believe that's it for now - please let me know if I have forgotten > anything else! Any > help or information (resources, references, companies etc) anyone can > provide will be > greatly appreciate. > > regards > > Maria Mejia > neurohistologist > Smith-Kettlewell Eye Res. Inst. > San Francisco, CA 94115 > Email: maria@ski.org From Kemlo.Rogerson <@t> elht.nhs.uk Mon Dec 6 02:21:53 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] dioxane chemical questions[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B57E@bhrv-nt-11.bhrv.nwest.nhs.uk> You have good health insurance? In a union? -----Original Message----- From: Maria Mejia [mailto:maria@ski.org] Sent: 03 December 2004 23:16 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] dioxane chemical questions[Scanned] Hello Everyone, in the next few weeks, we plan to use very small amounts of (dioxane) to coat coverslip - 10ml -20ml at one time - 2-4 times /month. In previous experiments, the PI has tried other chemical substitutes - without success. Without the use of dioxane, a protein called conA that attaches cells to the glass will not bond properly with the coverslips and instead will wash away during testing. This question is for anyone (past or current) using dioxane, please advise, on the following procedure: I understand in order to prevent peroxidizable compounds in the dioxane, it is necessary to store the liquid material - inerting the atmosphere in storage containers with nitrogen to inhibit peroxide formation. >When using dioxane, is there another alternative to storing in nitrogen or argon atmosphere? For example, adding an inert high boiling substance like mineral oil to prevent the peroxide from concentrating to a dangerous level? Has anyone tried this? >What type of containers (metal or plastic) should be used for cryogenic process? >What type of container should be used in disposal of dioxane? Polyethylene plastic? > Can I assume that -20C is a good temperature for storage? > How long should I keep working dioxane? I read 3-12 months. > How long should I keep waste dioxane waiting for disposal pick-up? > Disposal of peroxides - should I dilute with water - what concentration of H20? > How can I detect & determine peroxides? I notice in the Laboratory Safety that they sell peroxide check paper strips. Does anyone know if these strips are reliable and accurate? > How often should we have the fume hood where dioxane will be use...checked? Every six months? Once a year (currently doing)? Nitrile (heavy weight - 8mil-12 mil) and butyl gloves will be used. I believe that's it for now - please let me know if I have forgotten anything else! Any help or information (resources, references, companies etc) anyone can provide will be greatly appreciate. regards Maria Mejia neurohistologist Smith-Kettlewell Eye Res. Inst. San Francisco, CA 94115 Email: maria@ski.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kmerriam2003 <@t> yahoo.com Mon Dec 6 06:45:45 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Mouse Paws and Gayle's Knowledge in a Book In-Reply-To: <20041203185612.92300.qmail@web54708.mail.yahoo.com> Message-ID: <20041206124545.73202.qmail@web52509.mail.yahoo.com> I agree - Gayle is the histology GURU for all things related to animal histology!!!! Kim Phillip Huff wrote: How many people think that Gayle should write a book describing her vast (correction - extremely vast) knowledge of histochemistry and tissue sample prep? Phil --------------------------------- Do you Yahoo!? Send holiday email and support a worthy cause. Do good. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Meet the all-new My Yahoo! – Try it today! From mrsgbd2001 <@t> yahoo.com Mon Dec 6 07:14:06 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Gayle's Knowledge in a Book In-Reply-To: <20041206124545.73202.qmail@web52509.mail.yahoo.com> Message-ID: <20041206131406.68708.qmail@web52703.mail.yahoo.com> I also agree, Gayle has helped me on numerous occassions with her "extremely vast" knowledge. Gareth Davis Kim Merriam wrote: I agree - Gayle is the histology GURU for all things related to animal histology!!!! Kim Phillip Huff wrote: How many people think that Gayle should write a book describing her vast (correction - extremely vast) knowledge of histochemistry and tissue sample prep? Phil --------------------------------- Do you Yahoo!? Send holiday email and support a worthy cause. Do good. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Meet the all-new My Yahoo! – Try it today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Read only the mail you want - Yahoo! Mail SpamGuard. From Kemlo.Rogerson <@t> elht.nhs.uk Mon Dec 6 07:25:57 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Mouse Paws and Gayle's Knowledge in a Book[Scanned ] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B589@bhrv-nt-11.bhrv.nwest.nhs.uk> Aren't we animals too? That means she omnipotent. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: 06 December 2004 12:46 To: Phillip Huff; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Mouse Paws and Gayle's Knowledge in a Book[Scanned] I agree - Gayle is the histology GURU for all things related to animal histology!!!! Kim Phillip Huff wrote: How many people think that Gayle should write a book describing her vast (correction - extremely vast) knowledge of histochemistry and tissue sample prep? Phil --------------------------------- Do you Yahoo!? Send holiday email and support a worthy cause. Do good. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Meet the all-new My Yahoo! - Try it today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Myri37 <@t> aol.com Mon Dec 6 08:56:34 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] eosin hematoxylin plastic stain Message-ID: <722C7A2B.6A1E1039.0005167B@aol.com> hi i would like to stain plastic sections about 50 um thikness with eosine/ hematoxyline stain, wich hematoxyline should i use ? : hematoxyline solution harris modified (7g/L) or Hematoxyline mayer's (1g/l)? i found some protocols with phloxine B and eosin Y, and others with eosin only, do you have any information about these use ? thank you very much in advance myriam natural implant france From gcallis <@t> montana.edu Mon Dec 6 09:49:35 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:22 2005 Subject: [Histonet] Enough !! on Mouse Paws and Gayle's Knowledge in a Book In-Reply-To: <1030B679AD69D6119C3F00080210DD9D01B0B589@bhrv-nt-11.bhrv.n west.nhs.uk> References: <1030B679AD69D6119C3F00080210DD9D01B0B589@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: <6.0.0.22.1.20041206083505.01b3c288@gemini.msu.montana.edu> Enough! First of all, I doubt I will be doing a book, but thanks for the encouragement. You wrote: Aren't we animals too? That means she omnipotent. Kemlo Rogerson Second: Yes, we are animals and I have worked with MOST species, no ALL (hence omnipotent is not really a valid term here, and should be reserved for powers greater me - in my estimation) Third, let's get back to Histonet format, PLEASE! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Dec 6 10:23:05 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:22 2005 Subject: H&E on plastic, which plastic? Re: [Histonet] eosin hematoxylin plastic stain In-Reply-To: <722C7A2B.6A1E1039.0005167B@aol.com> References: <722C7A2B.6A1E1039.0005167B@aol.com> Message-ID: <6.0.0.22.1.20041206085108.01b6eca0@gemini.msu.montana.edu> Myriam, You did not say which plastic you are working with? I presume bone is undecalcified and embedded in methylmethacrylate? H&E an methylmethacrylate cannot be done successfully unless you remove ALL the plastic. You should read this article on why H &E is not going to work on bone embedded in methylmethacrylate sections. It also discussed considerations with Glycol methacrylate. First of all hematoxylin is an aqueous solution and second the molecule is high molecular weight. Methylmethacrylate is extremely hydrophobic with GMA only less so. The aqueous hematoxylin stain will not penetrate to the cells. If you etch the surface of bone in MMA with formic acid, it only removes the calcium from the bone, but does not remove plastic. RW Horobin, Staining plastic sections: a review of problems, explanations and possible solutions. J of Microscopy 131(2):173-186, August 1983. I do not know of anyone who is successful with an H&E stain on undecalcified, methylmethacrylate embedded bone section. IIf I recall, people who have tried H&E on bone in MMA get "muddy" very poor staining results. Neil Hand did VERY thin undecalcified bone MMA embedded sections at 2 um, but he removed ALL plastic before H&E staining. Calcium in bone stains very blue. I know one person who developed an H&E "look alike" stain (toluidine blue with basic fuchsin) on undecalcified methylmethacrylate bone sections. This provided contrast but not a true tissue-chemical interaction by eosin as with eosinophils or eosinophilia. There is nice contrast with bone cells, bone, cartilage, soft tissues, etc. This can be done on thick sections with surface staining - after formic acid etching of surface. Very nice stain, and plastic did not have to be removed. H&E is very successful in decalcified bone sections, not sure about 50 um thick though, and we do a progressive hematoxylin on decalcified bone but NOT Mayers, you want to get deeper staining of the bone along with other soft tissue components. In Bancroft and Stevens 4th Edition, Theory and Practice of Histological Technique, there are pictures of Mayers hematoxylin on decalcified bone next to Ehrlichs hematoxylin on an adjacent section. The Mayers looks washed out, the cartilage stains poorly as compared to Ehrlichs. We have done Ehrlichs hematoxylin on decalcifed bone sections but on 5 - 10 um thickness. 50 um sections, if the stain could penetrate all the way through the undecalcified bone and even with undecalcified will probably mask a lot of what you want to see. As far as eosin versus eosin phloxine, we prefer the latter for decalcified bone sections. 4, you wrote: >hi >i would like to stain plastic sections about 50 um thikness with eosine/ >hematoxyline stain, wich hematoxyline should i use ? : >hematoxyline solution harris modified (7g/L) or Hematoxyline mayer's (1g/l)? >i found some protocols with phloxine B and eosin Y, and others with eosin >only, do you have any information about these use ? >thank you very much in advance >myriam >natural implant >france > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From weneng2004 <@t> yahoo.com Mon Dec 6 18:17:58 2004 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] CD31 (PECAM) Message-ID: <20041207001758.85434.qmail@web53405.mail.yahoo.com> Hello, I was asked to do PECAM IHC on paraffin embedded mouse tissue. Could anybody tell me which antibody is the best? Thanks in advance! Wen --------------------------------- Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. From liz <@t> premierlab.com Mon Dec 6 18:48:28 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] CD31 (PECAM) In-Reply-To: <20041207001758.85434.qmail@web53405.mail.yahoo.com> Message-ID: <000201c4dbf6$77bbf160$76d48a80@AMY> Santa Cruz has a goat polyclonal that works great in FFPE mouse tissue with HIER retrieval. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of wen eng Sent: Monday, December 06, 2004 5:18 PM To: histonet Subject: [Histonet] CD31 (PECAM) Hello, I was asked to do PECAM IHC on paraffin embedded mouse tissue. Could anybody tell me which antibody is the best? Thanks in advance! Wen --------------------------------- Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Edward.Crisp <@t> leica-microsystems.com Mon Dec 6 19:00:45 2004 From: Edward.Crisp <@t> leica-microsystems.com (Edward.Crisp@leica-microsystems.com) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Edward Crisp/GBMIK/LMSCentral/Leica is out of the office. Message-ID: I will be out of the office starting 06/12/2004 and will not return until 10/12/2004. I am on Annual leave from Moday 6th December till Friday 10th December. I will be collecting my emails during this period, however, should you have any urgent requirements please feel free to call me on my mobile. ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From WWmn916 <@t> aol.com Mon Dec 6 21:52:03 2004 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] HIPPA and slides Message-ID: <141.3a8cdfcb.2ee682e3@aol.com> Does anyone know if patient names can go on slides in addition to accession numbers? Doing so seems like it would go against HIPPA law. Thanks Deb From bettina.hutz <@t> orionpharma.com Tue Dec 7 00:43:17 2004 From: bettina.hutz <@t> orionpharma.com (bettina.hutz@orionpharma.com) Date: Fri Sep 16 15:24:23 2005 Subject: H&E on plastic, which plastic? Re: [Histonet] eosinhematoxyli n plastic stain Message-ID: <243E1A79AF5D89438C70513DBADAA4F0019E4008@sfies-exchange1.orionnet.org> Hello, I have been doing H&E on bones embedded in methylmetacrylate, and it worked fine! I used hematoxylin after GILL and alcoholic Eosin. Tina -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Monday, December 06, 2004 6:23 PM To: Myri37@aol.com; Histonet@lists.utsouthwestern.edu Subject: H&E on plastic, which plastic? Re: [Histonet] eosinhematoxylin plastic stain Myriam, You did not say which plastic you are working with? I presume bone is undecalcified and embedded in methylmethacrylate? H&E an methylmethacrylate cannot be done successfully unless you remove ALL the plastic. You should read this article on why H &E is not going to work on bone embedded in methylmethacrylate sections. It also discussed considerations with Glycol methacrylate. First of all hematoxylin is an aqueous solution and second the molecule is high molecular weight. Methylmethacrylate is extremely hydrophobic with GMA only less so. The aqueous hematoxylin stain will not penetrate to the cells. If you etch the surface of bone in MMA with formic acid, it only removes the calcium from the bone, but does not remove plastic. RW Horobin, Staining plastic sections: a review of problems, explanations and possible solutions. J of Microscopy 131(2):173-186, August 1983. I do not know of anyone who is successful with an H&E stain on undecalcified, methylmethacrylate embedded bone section. IIf I recall, people who have tried H&E on bone in MMA get "muddy" very poor staining results. Neil Hand did VERY thin undecalcified bone MMA embedded sections at 2 um, but he removed ALL plastic before H&E staining. Calcium in bone stains very blue. I know one person who developed an H&E "look alike" stain (toluidine blue with basic fuchsin) on undecalcified methylmethacrylate bone sections. This provided contrast but not a true tissue-chemical interaction by eosin as with eosinophils or eosinophilia. There is nice contrast with bone cells, bone, cartilage, soft tissues, etc. This can be done on thick sections with surface staining - after formic acid etching of surface. Very nice stain, and plastic did not have to be removed. H&E is very successful in decalcified bone sections, not sure about 50 um thick though, and we do a progressive hematoxylin on decalcified bone but NOT Mayers, you want to get deeper staining of the bone along with other soft tissue components. In Bancroft and Stevens 4th Edition, Theory and Practice of Histological Technique, there are pictures of Mayers hematoxylin on decalcified bone next to Ehrlichs hematoxylin on an adjacent section. The Mayers looks washed out, the cartilage stains poorly as compared to Ehrlichs. We have done Ehrlichs hematoxylin on decalcifed bone sections but on 5 - 10 um thickness. 50 um sections, if the stain could penetrate all the way through the undecalcified bone and even with undecalcified will probably mask a lot of what you want to see. As far as eosin versus eosin phloxine, we prefer the latter for decalcified bone sections. 4, you wrote: >hi >i would like to stain plastic sections about 50 um thikness with >eosine/ >hematoxyline stain, wich hematoxyline should i use ? : >hematoxyline solution harris modified (7g/L) or Hematoxyline mayer's (1g/l)? >i found some protocols with phloxine B and eosin Y, and others with eosin >only, do you have any information about these use ? >thank you very much in advance >myriam >natural implant >france > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Fri Dec 3 17:27:58 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] vacation Message-ID: Item Type: Note Date: Monday, 6 Dec 2004 I am on vacation 12/6-12/10...have a good week From Kemlo.Rogerson <@t> elht.nhs.uk Tue Dec 7 02:25:06 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Ventana vs Dako[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B590@bhrv-nt-11.bhrv.nwest.nhs.uk> I'm sorry to labour this thread, especially as I have received so much good advice, but it only served to generate further questions. If I were to merge two relatively large Immuno Labs to give a total number of slides of about 12,000 per annum would the cost per test go down with either system, remain the same or go up. Which one, Ventana or Dako, would provide the 'economies of scale'? Which platform performs best in NEQAS and can new procedures be implemented quickly on either platform. I mean if a new antibody were developed could it be made available quickly on either system? Which company is more inventive/ innovative with its products? What about support? Anyone want to buy a second hand Dako or Ventana? Which one would you prefer and why? Unfortunately if you are too persuasive with your argument only the one you don't want becomes available. Why do Immuno Lab Staff still cook their lunch in a pressure cooker? Especially as they also seem to have a microwave too, is that for the pies? From Ian.Ward <@t> nottingham.ac.uk Tue Dec 7 03:24:13 2004 From: Ian.Ward <@t> nottingham.ac.uk (Ian Ward) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] H and E staining Message-ID: Hi all I could use some advice, I have been trying to H and E stain some mouse sections of the lungs, brain, spleen, liver, kidney, pancreas and heart. i have used the following method:- Xlylene 5minutes Xlylene 5minutes 100% IMS 5minutes 100% IMS 5minutes 90% IMS 5minutes 70% IMS 5minutes 50% IMS 5minutes running tap water 5minutes Filtered Haematoxylin 5minutes Quick wash in running tap water Dip 20 times in Acid Alcohol Quick wash in running tap water Few seconds in Lithium Carbonate 1% Eosin 5minutes Quick rince in running tap water Take quickly though alcohols to xylene again Mount My problem is that for some reason very little of the Eosin can be seen on the sections when I come to view them. Any ideas? Cheers Ian Ian Ward Senior Technician E Floor The Medical School QMC Nottingham NG7 2UH Tel: 0115 9249924 Ext 36179 This message has been scanned but we cannot guarantee that it and any attachments are free from viruses or other damaging content: you are advised to perform your own checks. Email communications with the University of Nottingham may be monitored as permitted by UK legislation. From lpwenk <@t> sbcglobal.net Tue Dec 7 03:53:34 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] H and E staining References: Message-ID: <002d01c4dc42$9f4d1dc0$0c21d445@domainnotset.invalid> Few questions: 1. Directly from lithium carbonate to eosin? There should be a running water rinse between. 2-5 minutes. Going from bluing agent directly to eosin would change the pH of the eosin. 2. What's the pH of the eosin? Should be around 4-4.5, acidified with acetic acid. 3. Eosin solvent? 70% alcohol works for us. 4. Water rinse after eosin? Try going directly into 95% alcohol - 2 changes, 10 seconds with vigorous dipping, then to 2-3 changes of abs. alcohol for the same, then xylene several changes. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48037 ----- Original Message ----- From: "Ian Ward" To: Sent: Tuesday, December 07, 2004 4:24 AM Subject: [Histonet] H and E staining Hi all I could use some advice, I have been trying to H and E stain some mouse sections of the lungs, brain, spleen, liver, kidney, pancreas and heart. i have used the following method:- Xlylene 5minutes Xlylene 5minutes 100% IMS 5minutes 100% IMS 5minutes 90% IMS 5minutes 70% IMS 5minutes 50% IMS 5minutes running tap water 5minutes Filtered Haematoxylin 5minutes Quick wash in running tap water Dip 20 times in Acid Alcohol Quick wash in running tap water Few seconds in Lithium Carbonate 1% Eosin 5minutes Quick rince in running tap water Take quickly though alcohols to xylene again Mount My problem is that for some reason very little of the Eosin can be seen on the sections when I come to view them. Any ideas? Cheers Ian Ian Ward Senior Technician E Floor The Medical School QMC Nottingham NG7 2UH Tel: 0115 9249924 Ext 36179 This message has been scanned but we cannot guarantee that it and any attachments are free from viruses or other damaging content: you are advised to perform your own checks. Email communications with the University of Nottingham may be monitored as permitted by UK legislation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Tue Dec 7 03:57:50 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] H and E staining Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0B8EBE@sjhaexc02.sjha.org> I would wash after the lithium carbonate. Is your eosin alcoholic? I would make it in 70% alcohol. Then dip slides in 80% alcohol between the lithium and eosin. Then go directly into 95% alcohol, absolute, xylene as usual. Joyce ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ian Ward Sent: Tue 12/7/2004 4:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] H and E staining Hi all I could use some advice, I have been trying to H and E stain some mouse sections of the lungs, brain, spleen, liver, kidney, pancreas and heart. i have used the following method:- Xlylene 5minutes Xlylene 5minutes 100% IMS 5minutes 100% IMS 5minutes 90% IMS 5minutes 70% IMS 5minutes 50% IMS 5minutes running tap water 5minutes Filtered Haematoxylin 5minutes Quick wash in running tap water Dip 20 times in Acid Alcohol Quick wash in running tap water Few seconds in Lithium Carbonate 1% Eosin 5minutes Quick rince in running tap water Take quickly though alcohols to xylene again Mount My problem is that for some reason very little of the Eosin can be seen on the sections when I come to view them. Any ideas? Cheers Ian Ian Ward Senior Technician E Floor The Medical School QMC Nottingham NG7 2UH Tel: 0115 9249924 Ext 36179 This message has been scanned but we cannot guarantee that it and any attachments are free from viruses or other damaging content: you are advised to perform your own checks. Email communications with the University of Nottingham may be monitored as permitted by UK legislation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Kemlo.Rogerson <@t> elht.nhs.uk Tue Dec 7 04:56:56 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] H and E staining[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B598@bhrv-nt-11.bhrv.nwest.nhs.uk> Eosin likes Calcium for a reason I can't remember. An old Senior Chief who taught me always 'mordanted' in calcium formate before putting into aqueous eosin that contained calcium ions too. I concur over the washing after lithium as it removes crystals and it's always nice to be clean and manic rather than dirty and depressed. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: 07 December 2004 09:58 To: Ian Ward; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H and E staining[Scanned] I would wash after the lithium carbonate. Is your eosin alcoholic? I would make it in 70% alcohol. Then dip slides in 80% alcohol between the lithium and eosin. Then go directly into 95% alcohol, absolute, xylene as usual. Joyce ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ian Ward Sent: Tue 12/7/2004 4:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] H and E staining Hi all I could use some advice, I have been trying to H and E stain some mouse sections of the lungs, brain, spleen, liver, kidney, pancreas and heart. i have used the following method:- Xlylene 5minutes Xlylene 5minutes 100% IMS 5minutes 100% IMS 5minutes 90% IMS 5minutes 70% IMS 5minutes 50% IMS 5minutes running tap water 5minutes Filtered Haematoxylin 5minutes Quick wash in running tap water Dip 20 times in Acid Alcohol Quick wash in running tap water Few seconds in Lithium Carbonate 1% Eosin 5minutes Quick rince in running tap water Take quickly though alcohols to xylene again Mount My problem is that for some reason very little of the Eosin can be seen on the sections when I come to view them. Any ideas? Cheers Ian Ian Ward Senior Technician E Floor The Medical School QMC Nottingham NG7 2UH Tel: 0115 9249924 Ext 36179 This message has been scanned but we cannot guarantee that it and any attachments are free from viruses or other damaging content: you are advised to perform your own checks. Email communications with the University of Nottingham may be monitored as permitted by UK legislation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Jackie.O'Connor <@t> abbott.com Tue Dec 7 06:41:53 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] CD31 (PECAM) Message-ID: I use Santa Cruz SC1506 1:750 with 6.1 pH HIER in a standard SABC protocol. JO'C wen eng Sent by: histonet-bounces@lists.utsouthwestern.edu 12/06/2004 06:17 PM To: histonet cc: Subject: [Histonet] CD31 (PECAM) Hello, I was asked to do PECAM IHC on paraffin embedded mouse tissue. Could anybody tell me which antibody is the best? Thanks in advance! Wen --------------------------------- Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From marytedo <@t> hotmail.com Tue Dec 7 08:32:33 2004 From: marytedo <@t> hotmail.com (María Teresa Domínguez) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] H and E staining In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E0B8EBE@sjhaexc02.sjha.org> Message-ID: Is your Eosin Alcoholic? that is the question... I usually dip the slides in 95% alcohol before Eosin,if it's alcoholic. After the eosin,95%alcohol(2 washes) absulute alcohol(2 washes),Xilene(2 washes), mount media as ususal. HT. Maria T Dominguez Patology Serv. H.R.R.G. Rio Grande, Tierra del Fuego Argentina. >From: "Weems, Joyce" >To: "Ian Ward" , >Subject: RE: [Histonet] H and E staining >Date: Tue, 7 Dec 2004 04:57:50 -0500 > >I would wash after the lithium carbonate. >Is your eosin alcoholic? I would make it in 70% alcohol. Then dip slides in 80% alcohol between the lithium and eosin. Then go directly into 95% alcohol, absolute, xylene as usual. >Joyce > >________________________________ > >From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ian Ward >Sent: Tue 12/7/2004 4:24 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] H and E staining > > > >Hi all > >I could use some advice, I have been trying to H and E stain some mouse sections of the lungs, brain, spleen, liver, kidney, pancreas and heart. i have used the following method:- >Xlylene 5minutes >Xlylene 5minutes >100% IMS 5minutes >100% IMS 5minutes >90% IMS 5minutes >70% IMS 5minutes >50% IMS 5minutes >running tap water 5minutes >Filtered Haematoxylin 5minutes >Quick wash in running tap water >Dip 20 times in Acid Alcohol >Quick wash in running tap water >Few seconds in Lithium Carbonate >1% Eosin 5minutes >Quick rince in running tap water >Take quickly though alcohols to xylene again >Mount > >My problem is that for some reason very little of the Eosin can be seen on the sections when I come to view them. Any ideas? > >Cheers > >Ian > > >Ian Ward >Senior Technician >E Floor >The Medical School >QMC >Nottingham >NG7 2UH >Tel: 0115 9249924 Ext 36179 > > >This message has been scanned but we cannot guarantee that it and any >attachments are free from viruses or other damaging content: you are >advised to perform your own checks. Email communications with the >University of Nottingham may be monitored as permitted by UK legislation. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. >Thank you. Saint Josephs Health System, Inc. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar [1]MSN Toolbar Get it now! References 1. http://g.msn.com/8HMAEN/2752??PS=47575 From Ianbernard <@t> netzero.com Tue Dec 7 08:41:03 2004 From: Ianbernard <@t> netzero.com (Ian) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Question concerning TTC staining In-Reply-To: <2D7FF27625F01942A2CBD6183F62694F08EAB7@PHSXMB3.partners.org> Message-ID: Yes, I was involved in a similar project like this and have some source references that I may be able to provide you. Thanks, SSgt I. R. Bernard 59th Clinical Research Squadron Lackland AFB San Antonio Texas -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lapointe, Nathalie,Ph.D. Sent: Sunday, December 05, 2004 2:18 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Question concerning TTC staining Hi, I'm a fellow student at the Brigham and Women's hospital (Boston, MA), and I need an information in order to complete a paper. TTC staining was used to measure infarct size in rats dying between 4 and 24 hours - does such staining can only be performed accurately when the heart is excised and stained immediately after death? If the hearts were allowed to sit for some variable time period in dead animals, does the measurements would not have been accurate and the data would be unreliable? Since infarct size was used as a criterion for grouping most of the other data sets, an inaccuracy introduced into this measurement may have affected nearly all other statistical comparisons presented in the paper. However from my experience, an 6-8-hour delay of TTC staining is reliable for evaluating heart infarct size in a rat. Does this probably is attributable to the slow deterioration of mitochondrial enzyme activity in nonischemic heart over this time period? The only paper that I found concerning that matter is not with the heart but with the brain (in rat): " Delayed triphenyltetrazolium chloride staining remains useful for evaluating cerebral infarct volume in a rat stroke model. Li F, Irie K, Answer MS, Fisher M. Department of Neurology, Memorial Health Care, Worcester, Massachusetts 01605, U.S.A." Does anybody have further references or info for me? Thank you very much, Dr Nathalie Lapointe _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Dec 7 08:47:58 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:23 2005 Subject: H&E on plastic, which plastic? Re: [Histonet] eosinhematoxyli n plastic stain In-Reply-To: <243E1A79AF5D89438C70513DBADAA4F0019E4008@sfies-exchange1.o rionnet.org> References: <243E1A79AF5D89438C70513DBADAA4F0019E4008@sfies-exchange1.orionnet.org> Message-ID: <6.0.0.22.1.20041207074614.01b2da88@gemini.msu.montana.edu> Are your sections thin, i.e. 5 um versus thick, 50 um? as Myriam indicated.. Do you remove the plastic BEFORE staining? Please provide some details Thanks At 11:43 PM 12/6/2004, you wrote: >Hello, > >I have been doing H&E on bones embedded in methylmetacrylate, and it worked >fine! I used hematoxylin after GILL and alcoholic Eosin. >Tina Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Jann.Nevard <@t> baycare.org Tue Dec 7 08:55:57 2004 From: Jann.Nevard <@t> baycare.org (Nevard, Jann) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] In need of Histologist Message-ID: <1CAFDB7DCC67D41189920008C7A44D720C64D83C@SJHEXCH01> Opportunity to work in 5th largest not-for-profit health care facility nation wide and the largest on the West Coast of Florida. Currently seeking a full time histologist licensed in the state of Florida. The shift is from 3am-11:30am. For further consideration please respond via email. Jann Nevard Regional Recruiter BayCare Heath System Ph: 727-519-1315 Toll Free: (866) 221-3222 option #1 Fax: 727-519-1323 jann.nevard@baycare.org From gcallis <@t> montana.edu Tue Dec 7 09:08:47 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] H and E staining In-Reply-To: References: Message-ID: <6.0.0.22.1.20041207074835.01b4eb00@gemini.msu.montana.edu> You do not need to do a water rinse AFTER the eosin unless you are working with aqueous eosin. This rinse is taking water soluble eosin out of the tissue. Stain in eosin, then go to 95% alcohol X 2 changes, 100% X 2, etc. The 95% alcohol will differentiate the eosin without removing it excessively. We stain murine FFPE tissues with eosin (alcoholic) for 1 minute and have often backed off that time if the sections are too red, a common, annoying problem. Hopefully, you do a good 1 minute running tap water rinse after Lithium carbonate to remove this alkaline solution. IF you alter the pH of eosin with this alkaline solution i.e. cation carryover, the eosin will not have a correct pH to do its job in pH 4.5 range. By quick washes, 1 minute is quick for us, we can go longer but NEVER less than that. Adequate rinsing makes sure hematoxylin, acid and alkaline solutions are rinsed away thoroughly. We add as 70% alcohol rinse before the eosin to do the following: 1) remove any residual cation carryover from LiCO3 - even after it's water rinse 2) equilibrate the section to the same concentration of alcohol as the eosin is dissolved in. If you need any guidelines, go to Richard Allan website staining manual and read about H&E staining - it is free and excellent how one can manipulate this stain to suit their needs i.e more contrast, deeper staining, etc, etc. At 02:24 AM 12/7/2004, you wrote: >Hi all > >I could use some advice, I have been trying to H and E stain some mouse >sections of the lungs, brain, spleen, liver, kidney, pancreas and heart. i >have used the following method:- >Xlylene 5minutes >Xlylene 5minutes >100% IMS 5minutes >100% IMS 5minutes >90% IMS 5minutes >70% IMS 5minutes >50% IMS 5minutes >running tap water 5minutes >Filtered Haematoxylin 5minutes >Quick wash in running tap water >Dip 20 times in Acid Alcohol >Quick wash in running tap water >Few seconds in Lithium Carbonate >1% Eosin 5minutes >Quick rince in running tap water >Take quickly though alcohols to xylene again >Mount > >My problem is that for some reason very little of the Eosin can be seen on >the sections when I come to view them. Any ideas? > >Cheers > >Ian > > >Ian Ward >Senior Technician >E Floor >The Medical School >QMC >Nottingham >NG7 2UH >Tel: 0115 9249924 Ext 36179 > > >This message has been scanned but we cannot guarantee that it and any >attachments are free from viruses or other damaging content: you are >advised to perform your own checks. Email communications with the >University of Nottingham may be monitored as permitted by UK legislation. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Janet.Bonner <@t> FLHOSP.ORG Tue Dec 7 09:37:40 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] HIPPA and slides Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB411E@fh2k093.fhmis.net> We are presently putting both on the label. It does seem against Hippa, but the slides shouldn't end up in too public a place. AND- there's allot of John Smiths out there, no way of telling which John Smith you've got. Having both name and surgical number on the label has diverted enough errors to know that this is definitely worth while. Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of WWmn916@aol.com Sent: Monday, December 06, 2004 10:52 PM To: histonet@pathology.swmed.edu Subject: [Histonet] HIPPA and slides Does anyone know if patient names can go on slides in addition to accession numbers? Doing so seems like it would go against HIPPA law. Thanks Deb _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From glorialimetti <@t> yahoo.com Tue Dec 7 09:44:28 2004 From: glorialimetti <@t> yahoo.com (Gloria Limetti) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] chicken embryo Message-ID: <20041207154428.31531.qmail@web52403.mail.yahoo.com> I am interested in information on "how to process" chicken embryos for paraffin and frozens and then onto immuno's and in-situ. I am also interested in finding an atlas worth purchsing for chicken embryo. Any help would be greatly appreciated, Gloria Limetti glorialimetti@yahoo.com __________________________________ Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. http://mobile.yahoo.com/maildemo From Kemlo.Rogerson <@t> elht.nhs.uk Tue Dec 7 10:27:49 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] HIPPA and slides[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5A5@bhrv-nt-11.bhrv.nwest.nhs.uk> We always, in the UK, I think put both on; it's part of our IQC. I suppose the choice is to be sued for mixing up the slides or be sued for releasing Patient's names; which is cheaper? -----Original Message----- From: Bonner, Janet [mailto:Janet.Bonner@FLHOSP.ORG] Sent: 07 December 2004 15:38 To: WWmn916@aol.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] HIPPA and slides[Scanned] We are presently putting both on the label. It does seem against Hippa, but the slides shouldn't end up in too public a place. AND- there's allot of John Smiths out there, no way of telling which John Smith you've got. Having both name and surgical number on the label has diverted enough errors to know that this is definitely worth while. Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of WWmn916@aol.com Sent: Monday, December 06, 2004 10:52 PM To: histonet@pathology.swmed.edu Subject: [Histonet] HIPPA and slides Does anyone know if patient names can go on slides in addition to accession numbers? Doing so seems like it would go against HIPPA law. Thanks Deb _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Tue Dec 7 13:30:40 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] chicken embryo In-Reply-To: <20041207154428.31531.qmail@web52403.mail.yahoo.com> References: <20041207154428.31531.qmail@web52403.mail.yahoo.com> Message-ID: <41B604E0.2030602@umdnj.edu> Here are some good atlases. Do not be put off by the fact that these books are "old", they are classics in the field and thus are timeless. All should be availble at any college or university library. "Early Embryology of the Chick" by Bradley Patten. "Fundamentals of Comparative Embryology of the Vertebrates" by Wm. Huettner. "Comparative Anatomy and Embryology" by Wm. Ballard. "An Introduction to Embryology" by Boris Balinsky. More of a text book than an atlas. As for techniques for preparing embryos, I have never done it but I am sure that someone on the list will help you out. Geoff Gloria Limetti wrote: >I am interested in information on "how to process" >chicken embryos for paraffin and frozens and then onto >immuno's and in-situ. I am also interested in finding >an atlas worth purchsing for chicken embryo. > >Any help would be greatly appreciated, > >Gloria Limetti >glorialimetti@yahoo.com > > > >__________________________________ >Do you Yahoo!? >Take Yahoo! Mail with you! Get it on your mobile phone. >http://mobile.yahoo.com/maildemo > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From RossS <@t> BaylorHealth.edu Tue Dec 7 10:40:12 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] HIPPA and slides[Scanned] Message-ID: I seem to remember either this being discussed on Histonet before or reading an article about this. Either way, my memory is that the discussion about HIPPA and patient names on slides and blocks concluded that HIPPA requires reasonable care be taken not to release information. The slides and blocks themselves aren't information until they are examined by a qualified person. Removing the name is not necessary prior to disposal. Having the name as well as surgical # on the block/slide is beneficial for proper identification. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kemlo Rogerson Sent: Tuesday, December 07, 2004 10:28 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] HIPPA and slides[Scanned] We always, in the UK, I think put both on; it's part of our IQC. I suppose the choice is to be sued for mixing up the slides or be sued for releasing Patient's names; which is cheaper? -----Original Message----- From: Bonner, Janet [mailto:Janet.Bonner@FLHOSP.ORG] Sent: 07 December 2004 15:38 To: WWmn916@aol.com; histonet@pathology.swmed.edu Subject: RE: [Histonet] HIPPA and slides[Scanned] We are presently putting both on the label. It does seem against Hippa, but the slides shouldn't end up in too public a place. AND- there's allot of John Smiths out there, no way of telling which John Smith you've got. Having both name and surgical number on the label has diverted enough errors to know that this is definitely worth while. Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of WWmn916@aol.com Sent: Monday, December 06, 2004 10:52 PM To: histonet@pathology.swmed.edu Subject: [Histonet] HIPPA and slides Does anyone know if patient names can go on slides in addition to accession numbers? Doing so seems like it would go against HIPPA law. Thanks Deb _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From histo <@t> bthosp.com Tue Dec 7 11:02:51 2004 From: histo <@t> bthosp.com (O'Brien, Sue) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Decedents from Nursing Homes Message-ID: <4D95C24EC0E4C84787A6919F1165B25E24B062@btmhems01.BTHOSP.INT> I was wondering how other hospitals handled receiving decedents from nursing homes. These are cases where the patient died at the nursing home, but there is a delay anticipated in the decedent being transferred to a Funeral Home. Currently, those cases are coming to the hospital morgue (for refrigeration). These are not Medical Examiner cases. Do other hospitals charge for this type of service? Would anyone be willing to share their policies with me? Thank-you, Sue O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May Court House, NJ 08210 Phone: 609-463-2449 e-mail: histo@bthosp.com From bill501 <@t> mindspring.com Tue Dec 7 12:06:48 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] HIPPA and slides[Scanned] In-Reply-To: <1030B679AD69D6119C3F00080210DD9D01B0B5A5@bhrv-nt-11.bhrv.nwest.nhs.uk> References: <1030B679AD69D6119C3F00080210DD9D01B0B5A5@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: At 4:27 PM +0000 12/7/04, Kemlo Rogerson wrote: >We always, in the UK, I think put both on; it's part of our IQC. I suppose >the choice is to be sued for mixing up the slides or be sued for releasing >Patient's names; which is cheaper? My philosophy is always to ask what is best for the patient from a pathology point of view - regardless of rules and regulations. I even make a statement on pap smear reports (conventional) that a patient's name is not on the slide (which is required by our state regulations). Bill From bpinkerton <@t> ccpathology.com Tue Dec 7 14:07:40 2004 From: bpinkerton <@t> ccpathology.com (Betty Pinkerton) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] SPECIAL STAINER Message-ID: <028E7300440A0A45A7968235F4FD5663035B5E@exchange.ccpathology.com> We are looking for a special stainer that will do Steiners along with other special stains. We have an Artisan right now and we are looking for a more dependable stainer. Thanks From cwscouten <@t> myneurolab.com Tue Dec 7 14:27:49 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Service Contract Features Message-ID: I hope nobody minds if I ask for some help planning a service contract for histology instruments. What should be in a practical service contract? Period? Preventive Maintenance? How often? Response time? What percentage of instrument cost is usually charged? Other features it should have? Should all electrical and mechanical parts be covered under the contract? Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com From aep10 <@t> cornell.edu Tue Dec 7 14:28:46 2004 From: aep10 <@t> cornell.edu (Anna Elisse Beaudin) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC on brain cryosections... help!! Message-ID: <2379.132.236.104.213.1102451326.squirrel@132.236.104.213> Hi all, I am having a TON of trouble with basic IHC on mouse brain cryostat sections! I have reviewed tons of protocols from the web and from diferent papers.. and I am constantly getting confused between protocols for fixed, paraffin-embedded sections, and those for cryosections. Here is an overview of the protocol I am currently using: -Collect cryosections on slides, leave at room temp 1-2 hours (as I'm collecting) - Quick-fix in buffered PFA (4%) for 10 minutes - Rinse 3-10' PBS - Block for endogenous peroxidase activity in 3% hydrogen peroxide in PBS - Rinse 3 * 10 min Tris-buffered saline - Block in 1%BSA, .3% triton X in TBS - in primary o/n at 4 C - Rinse 3 * 10 min TBS - in secondary 1 hr at room temp - Rinse 3* 10 min TBS -- avitin-biotin (vector elite kit) for half hour at room temp -- Rinse 2 * 10 min TBS, 1*10 min TB pH7.6 -- in DAB 2-3 minutes With this protocol, I get terrible background in my negative controls, and cannot identify any stain in my positive slides. The background I'm getting does not appear to be cellular at all -- there is just brown gunk everywhere. Can anyone point me in the right direction? Thank you so much in advance for your help! Best, Anna Beaudin Division of Nutritional Sciences Cornell University Ithaca, NY From JWEEMS <@t> sjha.org Tue Dec 7 14:28:45 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] SPECIAL STAINER Message-ID: <83AACDB0810528418AA106F9AE9B7F7E50764F@sjhaexc02.sjha.org> WE have the Artisan. We do Genta for Helicobacter. We use their Warthin Starry and then stain with Alcian blue and H&E off the stainer. It works great for us. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Betty Pinkerton Sent: Tuesday, December 07, 2004 3:08 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] SPECIAL STAINER We are looking for a special stainer that will do Steiners along with other special stains. We have an Artisan right now and we are looking for a more dependable stainer. Thanks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From cfavara <@t> niaid.nih.gov Tue Dec 7 15:38:27 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC on brain cryosections... help!! Message-ID: I am hoping some questions and suggestions will help you think in a way to solve this problem. I am making the assumption that the negative control you are speaking of is a No Primary control rather than a negative tissue control. If you are not doing a No Primary Control I would highly recommend one. If you have a no primary control with terrible background then the background is due to some other reagent or combination of reagents. You might want to give some thought to some additional controls to pin the cause of this background down. Possible causes would be: 1- secondary antibody concentration 2 - avidin biotin complex concentration 3- chromogen concentration or peroxide concentration I would not keep my frozen sections at RT for 1-2 hours I would air dry for 1-5 minutes and proceed with fixation. I you need time to collect as we all do, try keeping slides in -20 or -80 until ready to start or hold in PBS@4C following fixation. The difference between frozen and paraffin is that you need to deparaffinize and rehydrate paraffin embedded tissue prior to staining. Sometimes you need to do some sort of epitope unmasking but this can be done on frozen tissue as well. You did not say what antibody you are working with. If you are very new to IHC and frozen brain sections I would highly recommend staining for something like GFAP first and get some confidence using and understanding the technical aspects of the system. Hope this is helpful 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: Anna Elisse Beaudin [mailto:aep10@cornell.edu] Sent: Tuesday, December 07, 2004 1:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC on brain cryosections... help!! Hi all, I am having a TON of trouble with basic IHC on mouse brain cryostat sections! I have reviewed tons of protocols from the web and from diferent papers.. and I am constantly getting confused between protocols for fixed, paraffin-embedded sections, and those for cryosections. Here is an overview of the protocol I am currently using: -Collect cryosections on slides, leave at room temp 1-2 hours (as I'm collecting) - Quick-fix in buffered PFA (4%) for 10 minutes - Rinse 3-10' PBS - Block for endogenous peroxidase activity in 3% hydrogen peroxide in PBS - Rinse 3 * 10 min Tris-buffered saline - Block in 1%BSA, .3% triton X in TBS - in primary o/n at 4 C - Rinse 3 * 10 min TBS - in secondary 1 hr at room temp - Rinse 3* 10 min TBS -- avitin-biotin (vector elite kit) for half hour at room temp -- Rinse 2 * 10 min TBS, 1*10 min TB pH7.6 -- in DAB 2-3 minutes With this protocol, I get terrible background in my negative controls, and cannot identify any stain in my positive slides. The background I'm getting does not appear to be cellular at all -- there is just brown gunk everywhere. Can anyone point me in the right direction? Thank you so much in advance for your help! Best, Anna Beaudin Division of Nutritional Sciences Cornell University Ithaca, NY _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From poyandehnavi <@t> yahoo.com Tue Dec 7 16:01:11 2004 From: poyandehnavi <@t> yahoo.com (Poyan Dehnavi) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Question concerning stain production Message-ID: <20041207220111.8094.qmail@web42105.mail.yahoo.com> Hi all, im new to this list. Ive been searching this list for production of different stain types but didnt find anything. Perhaps my question has been taken up before or is so ridiculously simple that its laughable, so id like to apologise in advance if anyone is offended or disturbed by my questions. Id like to know if there is a possibility to, with simple, easy obtainable chemicals and raw materials produce the most common stains that are avaliable. Anything from Methylene Blue, Eosin, Haematoxylin, Orange G etc. I mean how did the old guys like Koch or Ehrlich produce them or stumble upon them anyway? Thanks in advance P. Dehnavi Stud. Med. Univ. Vienna Austria __________________________________ Do you Yahoo!? Yahoo! Mail - now with 250MB free storage. Learn more. http://info.mail.yahoo.com/mail_250 From katri <@t> cogeco.ca Tue Dec 7 17:53:07 2004 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Question concerning stain production References: <20041207220111.8094.qmail@web42105.mail.yahoo.com> Message-ID: <006e01c4dcb7$e6e1cc60$6a9a9618@Katri> Hi Poyan, Histology text books all give staining methods and how to make the reagents for most usual stains. Try this web site for instant information, if you don't have an access to a text book. You should try and get one or more. http://members.pgonline.com/~bryand/StainsFile/stain/stainindex.htm Good luck! Katri Katri Tuomala St.Joseph's Healthcare Hamilton, Ontario, Canada ----- Original Message ----- From: "Poyan Dehnavi" To: Sent: Tuesday, December 07, 2004 5:01 PM Subject: [Histonet] Question concerning stain production > Hi all, im new to this list. > > Ive been searching this list for production of > different stain types but didnt find anything. Perhaps > my question has been taken up before or is so > ridiculously simple that its laughable, so id like to > apologise in advance if anyone is offended or > disturbed by my questions. > > Id like to know if there is a possibility to, with > simple, easy obtainable chemicals and raw materials > produce the most common stains that are avaliable. > Anything from Methylene Blue, Eosin, Haematoxylin, > Orange G etc. I mean how did the old guys like Koch or > Ehrlich produce them or stumble upon them anyway? > > Thanks in advance > P. Dehnavi > Stud. Med. Univ. > Vienna Austria > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - now with 250MB free storage. Learn more. > http://info.mail.yahoo.com/mail_250 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From c.gorrie <@t> unsw.edu.au Tue Dec 7 17:53:25 2004 From: c.gorrie <@t> unsw.edu.au (Cath Gorrie) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC on brain cryosections... help!! In-Reply-To: <2379.132.236.104.213.1102451326.squirrel@132.236.104.213> Message-ID: on 8/12/04 7:28 AM, Anna Elisse Beaudin wrote....... > Hi all, > > I am having a TON of trouble with basic IHC on mouse brain cryostat > sections! I have reviewed tons of protocols from the web and from > diferent papers.. and I am constantly getting confused between > protocols for fixed, paraffin-embedded sections, and those for > cryosections. Here is an overview of the protocol I am currently > using: > > -Collect cryosections on slides, leave at room temp 1-2 hours (as I'm > collecting) > > - Quick-fix in buffered PFA (4%) for 10 minutes > - Rinse 3-10' PBS > - Block for endogenous peroxidase activity in 3% hydrogen peroxide in PBS > - Rinse 3 * 10 min Tris-buffered saline > - Block in 1%BSA, .3% triton X in TBS > - in primary o/n at 4 C > - Rinse 3 * 10 min TBS > - in secondary 1 hr at room temp > - Rinse 3* 10 min TBS > -- avitin-biotin (vector elite kit) for half hour at room temp > -- Rinse 2 * 10 min TBS, 1*10 min TB pH7.6 > -- in DAB 2-3 minutes > > With this protocol, I get terrible background in my negative controls, and > cannot identify any stain in my positive slides. The background I'm > getting does not appear to be cellular at all -- there is just brown gunk > everywhere. Can anyone point me in the right direction? Thank you so > much in advance for your help! > > Best, > Anna Beaudin > Division of Nutritional Sciences > Cornell University > Ithaca, NY > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From contact <@t> excaliburpathology.com Tue Dec 7 17:59:06 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Question concerning stain production Message-ID: <20041207235907.30867.qmail@web50310.mail.yahoo.com> Many of the early stains used in histology came from the textile industry to dye the fabrics different colors. They developed the aniline dyes to get brighter color. Many stains are naturally occurring ie: hematoxylin came from the logwood tree, but is now synthesized. I prefer to make my own stains because I know when they were made and have not been sitting on a manufaturers shelf until I purchase it. My Schiff's will last 2 years. Are you wanting to make the chemicals used in the stains or are you wanting to make the stains from scratch? Find a copy of Lillie's Histopathologic Technic and Practical Histochemistry for the chemical reaction theory behind the stains used to demonstrate the different tissue elements. But if you can't, any histology technique book will contain the procedures for the stains, such as Sheehan/Hrapchak's Theory and Practice of Histotechnology. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From contact <@t> excaliburpathology.com Tue Dec 7 18:04:06 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC on frozen brain ... help Message-ID: <20041208000406.33665.qmail@web50310.mail.yahoo.com> I am assuming you are sectioning mouse brain. What animal is your primary antibody made in? It should not be mouse, but any other species. Your primary should not be made in the same species your test tissue is from. UNLESS, you use a mouse-on-mouse kit for mouse tissue. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From c.gorrie <@t> unsw.edu.au Tue Dec 7 18:07:09 2004 From: c.gorrie <@t> unsw.edu.au (Cath Gorrie) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC on brain cryosections... help!! In-Reply-To: <2379.132.236.104.213.1102451326.squirrel@132.236.104.213> Message-ID: Take two. Anna, In general your protocol should work fine. There is nothing that should cause terrible background, even allowing for inter-user variation. Some thoughts to consider... Are you air drying in a clean dust free environment? Is the "dirt" on the slide away from your section? How thick are your sections? Do you gently agitate the sections during washes? - ie place in a jar of buffer rather than just streaming buffer over the surface. I agree that the concentrations of your reagents is a possible cause and I definitely agree that it would be sensible to trial your techniques with a well defined and robust antibody. GFAP is an ideal choice. This will give you a better idea of where the source of the problem is. Regards, Cathy ---------------------------------------------------------------------------- Catherine Gorrie School of Medical Sciences University of New South Wales Sydney, NSW ph: 02 9385 2462 fax: 02 9385 8016 e-mail: c.gorrie@unsw.edu.au ---------------------------------------------------------------------------- on 8/12/04 7:28 AM, Anna Elisse Beaudin wrote....... > Hi all, > > I am having a TON of trouble with basic IHC on mouse brain cryostat > sections! I have reviewed tons of protocols from the web and from > diferent papers.. and I am constantly getting confused between > protocols for fixed, paraffin-embedded sections, and those for > cryosections. Here is an overview of the protocol I am currently > using: > > -Collect cryosections on slides, leave at room temp 1-2 hours (as I'm > collecting) > > - Quick-fix in buffered PFA (4%) for 10 minutes > - Rinse 3-10' PBS > - Block for endogenous peroxidase activity in 3% hydrogen peroxide in PBS > - Rinse 3 * 10 min Tris-buffered saline > - Block in 1%BSA, .3% triton X in TBS > - in primary o/n at 4 C > - Rinse 3 * 10 min TBS > - in secondary 1 hr at room temp > - Rinse 3* 10 min TBS > -- avitin-biotin (vector elite kit) for half hour at room temp > -- Rinse 2 * 10 min TBS, 1*10 min TB pH7.6 > -- in DAB 2-3 minutes > > With this protocol, I get terrible background in my negative controls, and > cannot identify any stain in my positive slides. The background I'm > getting does not appear to be cellular at all -- there is just brown gunk > everywhere. Can anyone point me in the right direction? Thank you so > much in advance for your help! > > Best, > Anna Beaudin > Division of Nutritional Sciences > Cornell University > Ithaca, NY > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 8 02:47:33 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Question concerning stain production[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5A8@bhrv-nt-11.bhrv.nwest.nhs.uk> Well that's how we used to make my boy. Take one Logwood trunk, boil for 20 days over a low heat........... Do you mean make the stains up from the constituent dyes and chemicals, not actually make the dye eosin. If it's the former then any good old book like Lillie will tell you, If it's the latter, why? -----Original Message----- From: Poyan Dehnavi [mailto:poyandehnavi@yahoo.com] Sent: 07 December 2004 22:01 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Question concerning stain production[Scanned] Hi all, im new to this list. Ive been searching this list for production of different stain types but didnt find anything. Perhaps my question has been taken up before or is so ridiculously simple that its laughable, so id like to apologise in advance if anyone is offended or disturbed by my questions. Id like to know if there is a possibility to, with simple, easy obtainable chemicals and raw materials produce the most common stains that are avaliable. Anything from Methylene Blue, Eosin, Haematoxylin, Orange G etc. I mean how did the old guys like Koch or Ehrlich produce them or stumble upon them anyway? Thanks in advance P. Dehnavi Stud. Med. Univ. Vienna Austria __________________________________ Do you Yahoo!? Yahoo! Mail - now with 250MB free storage. Learn more. http://info.mail.yahoo.com/mail_250 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 8 02:48:12 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Question concerning stain production[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5A9@bhrv-nt-11.bhrv.nwest.nhs.uk> Late again, I concur... -----Original Message----- From: Paula Pierce [mailto:contact@excaliburpathology.com] Sent: 07 December 2004 23:59 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Question concerning stain production[Scanned] Many of the early stains used in histology came from the textile industry to dye the fabrics different colors. They developed the aniline dyes to get brighter color. Many stains are naturally occurring ie: hematoxylin came from the logwood tree, but is now synthesized. I prefer to make my own stains because I know when they were made and have not been sitting on a manufaturers shelf until I purchase it. My Schiff's will last 2 years. Are you wanting to make the chemicals used in the stains or are you wanting to make the stains from scratch? Find a copy of Lillie's Histopathologic Technic and Practical Histochemistry for the chemical reaction theory behind the stains used to demonstrate the different tissue elements. But if you can't, any histology technique book will contain the procedures for the stains, such as Sheehan/Hrapchak's Theory and Practice of Histotechnology. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 8 02:57:57 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] HIPPA and slides[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5AC@bhrv-nt-11.bhrv.nwest.nhs.uk> If we received an unnamed slide we would send it back to the Source. We need, usually, three items, Surname, Forename and DOB. Other info like date of smear, etc may also be added but that's a lot of writing. I remember having slides from two ladies with the same name, living in the same house but only with dissimilar DOB's; they were daughter and mother. -----Original Message----- From: Bill Blank [mailto:bill501@mindspring.com] Sent: 07 December 2004 18:07 To: histonet@pathology.swmed.edu Subject: RE: [Histonet] HIPPA and slides[Scanned] At 4:27 PM +0000 12/7/04, Kemlo Rogerson wrote: >We always, in the UK, I think put both on; it's part of our IQC. I suppose >the choice is to be sued for mixing up the slides or be sued for releasing >Patient's names; which is cheaper? My philosophy is always to ask what is best for the patient from a pathology point of view - regardless of rules and regulations. I even make a statement on pap smear reports (conventional) that a patient's name is not on the slide (which is required by our state regulations). Bill _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 8 03:59:46 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Agenda 4 change[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5AE@bhrv-nt-11.bhrv.nwest.nhs.uk> Anyone got any neat JD's and Person Specs for the Agenda for Change before we get shafted by Management? Any early implementers throw us a lifeline? Anyone done the JD's of BMS2's and 3's? From Inga.Hansson <@t> neuro.uu.se Wed Dec 8 04:54:53 2004 From: Inga.Hansson <@t> neuro.uu.se (Inga Hansson) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] methyl green Message-ID: Hi everyone, What exactly does methyl green stain? Thanks for any information ! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 8 08:03:34 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] methyl green[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5B1@bhrv-nt-11.bhrv.nwest.nhs.uk> Things green? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Inga Hansson [mailto:Inga.Hansson@neuro.uu.se] Sent: 08 December 2004 10:55 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] methyl green[Scanned] Hi everyone, What exactly does methyl green stain? Thanks for any information ! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Matthew_Frank <@t> URMC.Rochester.edu Wed Dec 8 08:08:02 2004 From: Matthew_Frank <@t> URMC.Rochester.edu (Frank, Matthew) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] methyl green Message-ID: Methyl Green C.I 42585 or Ethyl Green C.I. 42590: Can be used a nuclear stain (methyl green pyronin)to distinguish DNA from RNA. I use it as a counterstain in Altmann's mitochondria stain against acid fuchsin. -----Original Message----- From: Inga Hansson [mailto:Inga.Hansson@neuro.uu.se] Sent: Wednesday, December 08, 2004 5:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] methyl green Hi everyone, What exactly does methyl green stain? Thanks for any information ! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brett_connolly <@t> merck.com Wed Dec 8 08:31:08 2004 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] methyl green[Scanned] Message-ID: Inga, I checked this out some time ago. To be more specific, methyl green binds selectively to A-T DNA rich sequences in the major groove along the chromosome. I am not sure exactly what moieties in the DNA it binds...i.e. bases, sugars etc. See this reference, FEBS Letters 1993; 315(1):61-4. Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com -----Original Message----- From: Inga Hansson [mailto:Inga.Hansson@neuro.uu.se] Sent: 08 December 2004 10:55 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] methyl green[Scanned] Hi everyone, What exactly does methyl green stain? Thanks for any information ! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 8 08:36:51 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] methyl green[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5B3@bhrv-nt-11.bhrv.nwest.nhs.uk> Good section in 'Histopathology Technic and Practical Histochemistry RD Lillie and Harold M. Fullmer, 4th edition, Page 177 to 182.' Mr Lillie was the 'God' in my day and I maintain he still is! Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Frank, Matthew [mailto:Matthew_Frank@URMC.Rochester.edu] Sent: 08 December 2004 14:08 To: 'Inga Hansson'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] methyl green[Scanned] Methyl Green C.I 42585 or Ethyl Green C.I. 42590: Can be used a nuclear stain (methyl green pyronin)to distinguish DNA from RNA. I use it as a counterstain in Altmann's mitochondria stain against acid fuchsin. -----Original Message----- From: Inga Hansson [mailto:Inga.Hansson@neuro.uu.se] Sent: Wednesday, December 08, 2004 5:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] methyl green Hi everyone, What exactly does methyl green stain? Thanks for any information ! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 _______________________________________________ Histonet mailing list Histonet@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 jaimie_hk <@t> yahoo.co.uk Wed Dec 8 08:52:46 2004 From: jaimie_hk <@t> yahoo.co.uk (Jaimie Hoh) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC queries - Bowman's layer and Descemet's membrane Message-ID: <20041208145246.45904.qmail@web86905.mail.ukl.yahoo.com> Hi everyone, I am looking for help in Immunohistochemistry... Is there anyone out there who knows which antibody that would detect Descemet's membrane? I am also looking for another antibody that would detect Bowman's layer... Any help would be greatly appreciated. Thank you and seasons greetings to everyone! Jaimie Research Officer Singapore Eye Research Insitute ___________________________________________________________ Win a castle for NYE with your mates and Yahoo! Messenger http://uk.messenger.yahoo.com From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 8 09:04:25 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Microtomes[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5B5@bhrv-nt-11.bhrv.nwest.nhs.uk> Two Leica 2035 microtomes regularly serviced, one careful owner, mint condition. Anyone interested? Buyer to collect. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Jaimie Hoh [mailto:jaimie_hk@yahoo.co.uk] Sent: 08 December 2004 14:53 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC queries - Bowman's layer and Descemet's membrane[Scanned] Hi everyone, I am looking for help in Immunohistochemistry... Is there anyone out there who knows which antibody that would detect Descemet's membrane? I am also looking for another antibody that would detect Bowman's layer... Any help would be greatly appreciated. Thank you and seasons greetings to everyone! Jaimie Research Officer Singapore Eye Research Insitute ___________________________________________________________ Win a castle for NYE with your mates and Yahoo! Messenger http://uk.messenger.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Wed Dec 8 09:38:21 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Microtomes[Scanned] Message-ID: Kemlo asks: "Two Leica 2035 microtomes regularly serviced, one careful owner, mint condition. Anyone interested? Buyer to collect." They won't let you out huh? :-) Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk From gcallis <@t> montana.edu Wed Dec 8 09:51:32 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Microtomes[Scanned] In-Reply-To: <1030B679AD69D6119C3F00080210DD9D01B0B5B5@bhrv-nt-11.bhrv.n west.nhs.uk> References: <1030B679AD69D6119C3F00080210DD9D01B0B5B5@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: <6.0.0.22.1.20041208084929.01b32de8@gemini.msu.montana.edu> Hmm the way this reads as that the careful owner is in "mint condition"? Sorry I am not in UK, could use one of these - the microtome that is. Gayle Callis At 08:04 AM 12/8/2004, you wrote: >Two Leica 2035 microtomes regularly serviced, one careful owner, mint >condition. Anyone interested? Buyer to collect. > >Kemlo Rogerson >Cellular Pathology Manager >East Lancashire Hospitals NHS Trust >DD. 01254-294162 >Mobile 0774-9754194 > > >-----Original Message----- >From: Jaimie Hoh [mailto:jaimie_hk@yahoo.co.uk] >Sent: 08 December 2004 14:53 >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] IHC queries - Bowman's layer and Descemet's >membrane[Scanned] > >Hi everyone, > >I am looking for help in Immunohistochemistry... > >Is there anyone out there who knows which antibody >that would detect Descemet's membrane? > >I am also looking for another antibody that would >detect Bowman's layer... > > >Any help would be greatly appreciated. > >Thank you and seasons greetings to everyone! > >Jaimie >Research Officer >Singapore Eye Research Insitute > > > >___________________________________________________________ >Win a castle for NYE with your mates and Yahoo! Messenger >http://uk.messenger.yahoo.com > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Wed Dec 8 13:00:17 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC on brain cryosections... help!! In-Reply-To: <2379.132.236.104.213.1102451326.squirrel@132.236.104.213> References: <2379.132.236.104.213.1102451326.squirrel@132.236.104.213> Message-ID: <41B74F41.2010904@umdnj.edu> Hi Anna: Allowing unfixed sections to dry for two hours at room temp. is going to give you terrible morphology. Furthermore, loss of cellular integrity will allow the antigens you are looking for to diffuse out of their normal location, compounding your troubles. Solution? Fix by perfusion of the whole animal first with formalin/paraformaldehyde, then rinse out the fix, cryoprotect with sucrose, freeze and section. Then do your IHC. Also, you did not say how you are freezing the tissue. Tissue must be frozen very quickly for good morphology, putting it in the cryostat and waiting for it to freeze will be a disaster. If that is what you are doing, everything else is pointless. I suggest you try your IHC on well-fixed material first to see what good morphology and good localization looks like. Then move on to cryosections. Find someone in the Biology Dept./Vet School/Med School who has experience and spend an afternoon in their lab. Seeing how it is done will save you time in the long run. Geoff Anna Elisse Beaudin wrote: >Hi all, > > I am having a TON of trouble with basic IHC on mouse brain cryostat >sections! I have reviewed tons of protocols from the web and from >diferent papers.. and I am constantly getting confused between >protocols for fixed, paraffin-embedded sections, and those for >cryosections. Here is an overview of the protocol I am currently >using: > >-Collect cryosections on slides, leave at room temp 1-2 hours (as I'm >collecting) > >- Quick-fix in buffered PFA (4%) for 10 minutes >- Rinse 3-10' PBS >- Block for endogenous peroxidase activity in 3% hydrogen peroxide in PBS >- Rinse 3 * 10 min Tris-buffered saline >- Block in 1%BSA, .3% triton X in TBS >- in primary o/n at 4 C >- Rinse 3 * 10 min TBS >- in secondary 1 hr at room temp >- Rinse 3* 10 min TBS >-- avitin-biotin (vector elite kit) for half hour at room temp >-- Rinse 2 * 10 min TBS, 1*10 min TB pH7.6 >-- in DAB 2-3 minutes > >With this protocol, I get terrible background in my negative controls, and >cannot identify any stain in my positive slides. The background I'm >getting does not appear to be cellular at all -- there is just brown gunk >everywhere. Can anyone point me in the right direction? Thank you so >much in advance for your help! > >Best, >Anna Beaudin >Division of Nutritional Sciences >Cornell University >Ithaca, NY > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From MTitford <@t> aol.com Wed Dec 8 10:18:27 2004 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Making your own dyes? from scratch? Message-ID: <59.1c74b7ce.2ee88353@aol.com> P. Dehnavi in Austria asks, I think, if it is possible to make your own dyes from scratch. The answer is "yes" but you would need a well equipped laboratory. William Perkin prepared the color mauve from aniline in 1856 while trying to make quinine if I remember correctly (I was NOT there at the time!). To read about how dyes are made, a good place to start is the book, "The emergence of the German Dye Industry" by J. J. Beer. ( University of Illinois Press 1959). It gives a blow by blow account of how dyes were developed, how the German Government supported the research, and how companies we now think of as drug manufacturers got started (Ciba, Giegy, Bayer. etc.) A second book is "The History of Staining" by G. Clark & F. H. Kasten. (3ed edition, Williams & Wilkins 1983) Ehrlich's perception that dyes stained cells specifically lead to the evolution of chemotherapy. Mike Titford USA Pathology Mobile AL USA From contact <@t> excaliburpathology.com Wed Dec 8 10:31:53 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC queries Bowman's layer and Descemet's Membrane Message-ID: <20041208163153.10761.qmail@web50303.mail.yahoo.com> Hello I have been doing ophthalmic histology for 25 years. You do not need to do an immuno to demonstrate Descemet's membrane in the cornea. The simple PAS (periodic acid - Shiff's) stain will stain Descemet's a bright hot pink. It is routinely done with an H&E on recipient corneas from transplants. Bowman's, however, will not stain with PAS, but is the homogeneous layer right beneath the epithelium. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From cornettl <@t> hotmail.com Wed Dec 8 10:36:58 2004 From: cornettl <@t> hotmail.com (Lorraine Cornett) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Wright Giemsa staining on Bone Marrow Smears Message-ID: Would anyone like to share their procedure of Wright Giemsa stain on Bone Marrow smears? We have a procedure that has just recently been producing inconsistent staining, we would like to evaluate if there is a better/more economical/and/or easier method out there that will give us consistent "good" staining. Also if you would like to contribute any experiences with microwave tissue processing (what brand preferred) I would appreciate it. This is a new area that we may begin to explore. Thanks Lorraine Cornett, HT (ASCP) Highlands Pathology Blue Ridge Division, Kingsport, TN 423 224-5793 fax 423 224-5349 _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From cornettl <@t> hotmail.com Wed Dec 8 10:36:02 2004 From: cornettl <@t> hotmail.com (Lorraine Cornett) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Wright Giemsa staining on Bone Marrow Smears Message-ID: Would anyone like to share their procedure of Wright Giemsa stain on Bone Marrow smears? We have a procedure that has just recently been producing inconsistent staining, we would like to evaluate if there is a better/more economical/and/or easier method out there that will give us consistent "good" staining. Also if you would like to contribute any experiences with microwave tissue processing (what brand preferred) I would appreciate it. This is a new area that we may begin to explore. Thanks Lorraine Cornett, HT (ASCP) Highlands Pathology Blue Ridge Division, Kingsport, TN 423 224-5793 fax 423 224-5349 _________________________________________________________________ On the road to retirement? Check out MSN Life Events for advice on how to get there! http://lifeevents.msn.com/category.aspx?cid=Retirement From CEvanish <@t> mont-hosp.com Wed Dec 8 10:39:03 2004 From: CEvanish <@t> mont-hosp.com (Chris Evanish) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] ER staining problem Message-ID: I need your help. I use a Ventana ES and am having a real problem getting the ER antibody to work. It was always a great result, however in the last 6 months or so it has gotten worse to the point now where the stain is dead negative. I've tried different slides, different pressure cookers, different retrieval solution and even different ER antibodies. One from Ventana and one from Zymed. If you can help, or give me a few thing to try I would appreciate it. Thanks, Chris From contact <@t> excaliburpathology.com Wed Dec 8 10:51:07 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] methyl green Message-ID: <20041208165107.17775.qmail@web50303.mail.yahoo.com> Kemlo, of all the books in my personal collection, I treasure my Lillie's the most. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From jryan <@t> sleh.com Wed Dec 8 10:53:47 2004 From: jryan <@t> sleh.com (John Ryan) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] I'm sorry I can not reply immediately because I am out of the office, returning on Friday 12/10/04. Message-ID: I'm sorry I can not reply immediately because I am out of the office, returning on Friday 12/10/04. +++++CONFIDENTIALITY NOTICE+++++ The information in this e-mail may be confidential and/or privileged. If you are not the intended recipient or an authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments, if any, or the information contained herein is prohibited. If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail from your computer system. Thank you. From RossS <@t> BaylorHealth.edu Wed Dec 8 10:56:18 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] ER staining problem Message-ID: Are your slides cut fresh, or are they pre-cut days or weeks ahead? ER is one antibody some people have problems with if it is not cut fresh. Has something changed in the fixation? These are the only ideas I have on the subject right now. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Chris Evanish Sent: Wednesday, December 08, 2004 10:39 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] ER staining problem I need your help. I use a Ventana ES and am having a real problem getting the ER antibody to work. It was always a great result, however in the last 6 months or so it has gotten worse to the point now where the stain is dead negative. I've tried different slides, different pressure cookers, different retrieval solution and even different ER antibodies. One from Ventana and one from Zymed. If you can help, or give me a few thing to try I would appreciate it. Thanks, Chris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From Terry.Marshall <@t> rothgen.nhs.uk Wed Dec 8 10:57:06 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC queries Bowman's layer and Descemet's Membrane Message-ID: I am constantly amazed at the requests for ipox to stain things which are perfectly well recognised without it. But, that is an aside. A confession. I can never remember which b.... membrane is which, and only need the information every year or so. Do you have a mnemonic? (A mnemonic for the spelling of mnemonic would be useful too). 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: Paula Pierce [mailto:contact@excaliburpathology.com] Sent: 08 December 2004 16:32 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC queries Bowman's layer and Descemet's Membrane Hello I have been doing ophthalmic histology for 25 years. You do not need to do an immuno to demonstrate Descemet's membrane in the cornea. The simple PAS (periodic acid - Shiff's) stain will stain Descemet's a bright hot pink. It is routinely done with an H&E on recipient corneas from transplants. Bowman's, however, will not stain with PAS, but is the homogeneous layer right beneath the epithelium. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LBlack <@t> carilion.com Wed Dec 8 11:04:11 2004 From: LBlack <@t> carilion.com (Lisa Black) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Sakura Rapid Tissue Processor Message-ID: Hello All, The Histology Lab where I work became a recipient of the new Sakura Rapid Tissue Processor in August. We have been struggling with all the changes necessary with this technology to produce quality H & E slides. Is there anyone on this list who is also using this new machine? I would love to hear from others who are using this system. Thanks, Lisa Black Carilion Consolidated Laboratory Histology Manager LBLACK@CARILION.COM Roanoke, VA 540-985-4082 From tpmorken <@t> labvision.com Wed Dec 8 11:05:36 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Question concerning stain production[Scanned] Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA257F772@usca0082k08.labvision.apogent.com> Concerning how the oldtimers stumbled on dyes, you should look into the textile dyeing industry. This website is a good start. It tells the history of logwood and brazilwood, from which hematoxylin and brazilin dyes were made. http://waynesword.palomar.edu/ecoph4.htm 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: Poyan Dehnavi [mailto:poyandehnavi@yahoo.com] Sent: 07 December 2004 22:01 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Question concerning stain production[Scanned] Hi all, im new to this list. Ive been searching this list for production of different stain types but didnt find anything. Perhaps my question has been taken up before or is so ridiculously simple that its laughable, so id like to apologise in advance if anyone is offended or disturbed by my questions. Id like to know if there is a possibility to, with simple, easy obtainable chemicals and raw materials produce the most common stains that are avaliable. Anything from Methylene Blue, Eosin, Haematoxylin, Orange G etc. I mean how did the old guys like Koch or Ehrlich produce them or stumble upon them anyway? Thanks in advance P. Dehnavi Stud. Med. Univ. Vienna Austria From RITA.ANGEL <@t> UC.EDU Wed Dec 8 11:18:34 2004 From: RITA.ANGEL <@t> UC.EDU (Rita Angel) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] stain that binds mercury Message-ID: <5.1.0.14.2.20041208121519.00b3c940@ucmail3.uc.edu> Hi everyone, We have an investigator here that is interested in finding a stain that will localize mercury in sections of cicadas. Does anyone know of anything that will bind mercury in tissues? This is all the information I have at this point. Thank you for your help and expertise, Rita Angel, HT University of Cincinnati From Sherrobins <@t> aol.com Wed Dec 8 11:30:37 2004 From: Sherrobins <@t> aol.com (Sherrobins@aol.com) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Histotech Position Message-ID: <2DA6FE6A.4BA36C43.0CAFCB37@aol.com> Full time position available at an independent pathology laboratory. Great benefits, competitive salary with no weekend schedules. For inquires contact: Sherry Robinson, CT/HT (ASCP) Lab Manager 254.752.9621.ext 219 sherrobins@aol.com From gu.lang <@t> gmx.at Wed Dec 8 11:34:19 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Sakura Rapid Tissue Processor References: Message-ID: <00e601c4dd4c$26177120$eeeea8c0@server> Hi Lisa, It would be interesting, what changes you had to do? Did you get sections, that were more blue or red? Was cutting more difficult or easier? And what about grossing? Gudrun ----- Original Message ----- From: "Lisa Black" To: Sent: Wednesday, December 08, 2004 6:04 PM Subject: [Histonet] Sakura Rapid Tissue Processor > Hello All, > > The Histology Lab where I work became a recipient of the new Sakura > Rapid Tissue Processor in August. We have been struggling with all the > changes necessary with this technology to produce quality H & E slides. > Is there anyone on this list who is also using this new machine? I > would love to hear from others who are using this system. > > Thanks, > > Lisa Black > Carilion Consolidated Laboratory > Histology Manager > LBLACK@CARILION.COM > Roanoke, VA > 540-985-4082 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Felixdustin <@t> aol.com Wed Dec 8 12:02:16 2004 From: Felixdustin <@t> aol.com (Felixdustin@aol.com) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] HTL exam Message-ID: <100.846f769.2ee89ba8@aol.com> Hello: I am going to take HTL computer exam in about 10 days and I would like to talk to anyone who have taken this exam. If you can kindly taking you time to help me please reply to my email. Thank you! Emily From NLAPOINTE <@t> PARTNERS.ORG Wed Dec 8 12:24:21 2004 From: NLAPOINTE <@t> PARTNERS.ORG (Lapointe, Nathalie,Ph.D.) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] H and E vs. Masson Trichrome in cardiac injury Message-ID: <2D7FF27625F01942A2CBD6183F62694F08EABD@PHSXMB3.partners.org> > Hi, > > I found that website by hazard and I'm very impress! > > I have another questions for you: > > I'm currently exploring some hormones that may cause myocardial injury in > adrenalectomized rats. > The heart were stained with hematoxylin and eosin for light microscopic > analysis (x40). A scale from 0-4 was used to score the level of myocardial > injury. A score of 0 represented no damage. A score of 1 represented the > presence of myocytes demonstrating early necrotic changes such as nuclear > pyknosis or karylysis, and oesinophil staining of the cytoplasm associated > with the presence of scattered neutrophilic infiltrates. A score of 2 was > given when one clear area of necrosis (loss of myocardial cells with heavy > neutrophilic infiltrates) was observed. When two or more separate areas of > necrosis were found (implicating the presence of two different myocardial > infarctions in the same heart), but the areas localized and compromised less > then 50% of the ventricular wall, the hearts received a score of 3. A score of > 4 was assigned to hearts that demonstrated extensive areas of necrosis > compromising more than 50% of wither the left or the right ventricle. > > I'm looking for another way then the above score to measure cardiac injury. > Based to the fact that Hematoxylin is a dark purplish dye that will stain the > chromatin within the nucleus (therefore also monocyte infiltration), and that > Eosin is an orangish pink to red dye that stains the cytoplasmic material > including connective tissue and collagen (therefore doesn't stain necrotic > tissues), I was wondering if a software in which I could measure the contrast > of color related to injury vs. intact tissue exist? Does this same software > (if available) can also calculated the number of dark purplish dye (therefore > level of inflammation)? > > We also stained the same heart with Masson Trichrome. Do you think that > measuring the % of blue staining (collagen) in the whole heart at a > magnification of X4 is equivalent to H and E score described above? How can I > know if the blue staining with Masson Trichrome is fibrosis rather then > collagen or only red Sirus staining can measure fibrosis? > Thank you very much for all your help. Best, > Nathalie Lapointe > Brigham and Women's Hospital > LMRC #214 > 221 Longwood Ave > Boston, MA, USA > 02115 > (617) 515-5936 > > From galinadeyneko <@t> yahoo.com Wed Dec 8 12:36:52 2004 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Cryo Mbed Message-ID: <20041208183652.2658.qmail@web14525.mail.yahoo.com> Hi Dr.Scouten and histonetters, Does it mean that Cryo M Bed more "supportive" than OCT. I work with murine carotid arteries with atheroscl. plaque. .I fix them for 2 hours in 10 % formalin and embed in OCT, but I have to cut with T -28 C in microtome and mount on cool slides , otherwise I am not able to obtain good vessel' lumen and plaque scrolls up or falls down and still I receive 30% of distorted slices. Any advices will be appreciate. Galina Deyneko CV Department Novartis Cambridge MA 617-871-7613 --------------------------------- Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. From LuckG <@t> empirehealth.org Wed Dec 8 13:20:55 2004 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] ER staining problem Message-ID: Chris, What clone(s) are your two ER ab's? Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: Chris Evanish [mailto:CEvanish@mont-hosp.com] Sent: Wednesday, December 08, 2004 8:39 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] ER staining problem I need your help. I use a Ventana ES and am having a real problem getting the ER antibody to work. It was always a great result, however in the last 6 months or so it has gotten worse to the point now where the stain is dead negative. I've tried different slides, different pressure cookers, different retrieval solution and even different ER antibodies. One from Ventana and one from Zymed. If you can help, or give me a few thing to try I would appreciate it. Thanks, Chris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Wed Dec 8 13:30:29 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] stain that binds mercury Message-ID: I found a reference for Voight's modification of the Timm silver sulfide method that is specific for mercury. (Acta Histochem. 14:315, 1962). I found the reference in Lillie's Histopathologic Technic and Practical Histochemistry, Third Edition 1965. An oldie but a goodie. Good luck. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: Rita Angel [mailto:RITA.ANGEL@UC.EDU] Sent: Wednesday, December 08, 2004 11:19 AM To: histonet@pathology.swmed.edu Subject: [Histonet] stain that binds mercury Hi everyone, We have an investigator here that is interested in finding a stain that will localize mercury in sections of cicadas. Does anyone know of anything that will bind mercury in tissues? This is all the information I have at this point. Thank you for your help and expertise, Rita Angel, HT University of Cincinnati _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sherrobins <@t> aol.com Wed Dec 8 13:37:05 2004 From: Sherrobins <@t> aol.com (Sherrobins@aol.com) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Histotech Position Message-ID: <0095F9D4.02FDB465.0CAFCB37@aol.com> I had posted this position earlier in the day and mistakenly left out the locatin of the lab which is in Waco, Tx. Sherry Robinson, CT/HT(ASCP) 254.752.9621.ext 219 sherrobins@aol.com From akbitting <@t> geisinger.edu Wed Dec 8 14:26:55 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] QI study Message-ID: Just wondering if anyone out there has ever done a QI study on mislabelled slides? My QI coordinator and Lab Director want me to do this for 2005. I am trying to identify an acceptable threshhold. 100% would be great, but I am a realist. Any ideas? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From gcallis <@t> montana.edu Wed Dec 8 14:41:07 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Cryoembedding media woes, some advice and not a bash session In-Reply-To: <20041208183652.2658.qmail@web14525.mail.yahoo.com> References: <20041208183652.2658.qmail@web14525.mail.yahoo.com> Message-ID: <6.0.0.22.1.20041208132512.01afdad8@gemini.msu.montana.edu> I am not bashing anyone's cryomedia, but there are caveats about trying out new products without testing them out before the BIG project comes along. No matter which cryoembedding media you desire to use, one should ask for a sample or purchase the new product before filling lumens, lungs, small intestines to test the media holding quality before embarking on a whole, gigantic study. Reason why: We tested a new cryoembedding media with great hopes it would work for us. We use cryoembedding media to fill/distend intestinal lumen to visualize villi and Peyer's patches. We found the new media did NOT support the villi. We normally use OCT for this purpose, and found the OCT supported these fine tissue structures, allowing perfect sections with high quality morphology. The other new media did not work, sections were compressed, villi were chewed up and section quality was poor. It was later found the newer media works great with human tissues and was never tested on murine or animal tissue. I have no doubt the cryomedia was excellent for human work, but failed to work for murine special needs cryotomy. Advice it to test whatever is on the market as one or more cryoembedding media will show up that is superior for some things we do in the name of research. If you find the embedding media is excellent, better holding power - let us know - the info will be appreciated. At 11:36 AM 12/8/2004, you wrote: >Hi Dr.Scouten and histonetters, >Does it mean that Cryo M Bed more "supportive" than OCT. I work with >murine carotid arteries with atheroscl. plaque. .I fix them for 2 hours >in 10 % formalin and embed in OCT, but I have to cut with T -28 C in >microtome and mount on cool slides , otherwise I am not able to obtain >good vessel' lumen and plaque scrolls up or falls down and still I >receive 30% of distorted slices. Any advices will be appreciate. >Galina Deyneko >CV Department >Novartis Cambridge MA >617-871-7613 > > > > >--------------------------------- >Do you Yahoo!? > Take Yahoo! Mail with you! Get it on your mobile phone. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From jkiernan <@t> uwo.ca Wed Dec 8 14:47:35 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC queries Bowman's layer and Descemet's Membrane References: Message-ID: <41B76867.EF0F3C25@uwo.ca> "Marshall Terry Dr, Consultant Histopathologist" wrote: > (A mnemonic for the spelling of mnemonic would be useful too). It's mnemonic because it's to help with the mnemory. (Sometimes you hear it pronounced "pneumonic"; people who do that need a mnemonic to remember that there isn't a u in mnemonic.) -- ------------------------------- 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 cwscouten <@t> myneurolab.com Wed Dec 8 14:53:33 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC on brain cryosections... help!! Message-ID: I agree with Geoff McAuliffe in every detail, you need to perfuse, and you need to fast freeze. If perfusion is new to you, I suggest you review the manual and protocol posted on the left at the following link. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=Sacrifice+Equipment&idsubcategory=21 About the need for fast freezing, see http://www.myneurolab.com/global/Manuals/Tips%20and%20Techniques%20Freezing%20Artifact.pdf Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anna Elisse Beaudin Sent: Tuesday, December 07, 2004 2:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC on brain cryosections... help!! Hi all, I am having a TON of trouble with basic IHC on mouse brain cryostat sections! I have reviewed tons of protocols from the web and from diferent papers.. and I am constantly getting confused between protocols for fixed, paraffin-embedded sections, and those for cryosections. Here is an overview of the protocol I am currently using: -Collect cryosections on slides, leave at room temp 1-2 hours (as I'm collecting) - Quick-fix in buffered PFA (4%) for 10 minutes - Rinse 3-10' PBS - Block for endogenous peroxidase activity in 3% hydrogen peroxide in PBS - Rinse 3 * 10 min Tris-buffered saline - Block in 1%BSA, .3% triton X in TBS - in primary o/n at 4 C - Rinse 3 * 10 min TBS - in secondary 1 hr at room temp - Rinse 3* 10 min TBS -- avitin-biotin (vector elite kit) for half hour at room temp -- Rinse 2 * 10 min TBS, 1*10 min TB pH7.6 -- in DAB 2-3 minutes With this protocol, I get terrible background in my negative controls, and cannot identify any stain in my positive slides. The background I'm getting does not appear to be cellular at all -- there is just brown gunk everywhere. Can anyone point me in the right direction? Thank you so much in advance for your help! Best, Anna Beaudin Division of Nutritional Sciences Cornell University Ithaca, NY _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed Dec 8 15:07:35 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] stain that binds mercury References: Message-ID: <41B76D17.833C7B67@uwo.ca> Dear Rita, There are now more sensitive and selective modifications for staining mercury with Timm's sulphide-silver method. Method: Danscher G & Rungby J 1986. Histochem J 18:109-114. Some aplications: Arvidson B 1992. Muscle & Nerve 15:1089-1094. Pamphlett R & Waley P 1996. J Neurol Sci 135:63-67. Pamphlett R & Kum-Jew S 2001. Neurotoxicol Teratol 23:191-196. There may be more recent variations of the method, probably with Danscher as chief author. Try Google or Pubmed - or a cited ref search with Web of Science if that is available to you. It should be, at a big place like U of C. -- ------------------------------- 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 _______________________________ "GUTIERREZ, JUAN" wrote: > > I found a reference for Voight's modification of the Timm silver sulfide method that is specific for mercury. (Acta Histochem. 14:315, 1962). > I found the reference in Lillie's Histopathologic Technic and Practical Histochemistry, Third Edition 1965. An oldie but a goodie. Good luck. > > Juan C. Gutierrez, HT(ASCP) > Histology Laboratory Supervisor > (210)704-2533 > > My opinions are my own and do not reflect those of my employer. Long live free speech! > > -----Original Message----- > From: Rita Angel [mailto:RITA.ANGEL@UC.EDU] > Sent: Wednesday, December 08, 2004 11:19 AM > To: histonet@pathology.swmed.edu > Subject: [Histonet] stain that binds mercury > > Hi everyone, > > We have an investigator here that is interested in finding a stain that > will localize mercury in sections of cicadas. Does anyone know of anything > that will bind mercury in tissues? This is all the information I have at > this point. > > Thank you for your help and expertise, > > Rita Angel, HT > University of Cincinnati > > _______________________________________________ > Histonet mailing list > Histonet@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 fmonson <@t> wcupa.edu Wed Dec 8 15:14:01 2004 From: fmonson <@t> wcupa.edu (Monson, Frederick ) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] stain that binds mercury Message-ID: Using the following search string: "autometallography mercury pdf", I found a surfeit of information and protocols on google. However, the article by Danscher Journal of Histochemistry and Cytochemistry, Vol. 48, 1503-1510, November 2000 will also be very useful. You should be able to download it in PDF form directly since I can't imagine the UC lacks a subscription. Good luck, Fred Monson Frederick C. Monson, PhD Light, Electron, X-Ray and Scanning Probe Imaging and Analysis Center (LEXSPIAC) Large Scientific Instrument Core West Chester University S. Church St. and W. Rosedale Ave. West Chester, PA, 19320 610-738-0437 fmonson@wcupa.edu URL: http://darwin.wcupa.edu/casi -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rita Angel Sent: Wednesday, December 08, 2004 12:19 PM To: histonet@pathology.swmed.edu Subject: [Histonet] stain that binds mercury Hi everyone, We have an investigator here that is interested in finding a stain that will localize mercury in sections of cicadas. Does anyone know of anything that will bind mercury in tissues? This is all the information I have at this point. Thank you for your help and expertise, Rita Angel, HT University of Cincinnati _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From galinadeyneko <@t> yahoo.com Wed Dec 8 16:39:03 2004 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Re: Cryoembedding media woes, some advice and not a bash session In-Reply-To: <6.0.0.22.1.20041208132512.01afdad8@gemini.msu.montana.edu> Message-ID: <20041208223903.58937.qmail@web14521.mail.yahoo.com> Hi Gayle, I appreciate your advice. I always read with great interest your notes on Histonet. Galina Deyneko Gayle Callis wrote: I am not bashing anyone's cryomedia, but there are caveats about trying out new products without testing them out before the BIG project comes along. No matter which cryoembedding media you desire to use, one should ask for a sample or purchase the new product before filling lumens, lungs, small intestines to test the media holding quality before embarking on a whole, gigantic study. Reason why: We tested a new cryoembedding media with great hopes it would work for us. We use cryoembedding media to fill/distend intestinal lumen to visualize villi and Peyer's patches. We found the new media did NOT support the villi. We normally use OCT for this purpose, and found the OCT supported these fine tissue structures, allowing perfect sections with high quality morphology. The other new media did not work, sections were compressed, villi were chewed up and section quality was poor. It was later found the newer media works great with human tissues and was never tested on murine or animal tissue. I have no doubt the cryomedia was excellent for human work, but failed to work for murine special needs cryotomy. Advice it to test whatever is on the market as one or more cryoembedding media will show up that is superior for some things we do in the name of research. If you find the embedding media is excellent, better holding power - let us know - the info will be appreciated. At 11:36 AM 12/8/2004, you wrote: >Hi Dr.Scouten and histonetters, >Does it mean that Cryo M Bed more "supportive" than OCT. I work with >murine carotid arteries with atheroscl. plaque. .I fix them for 2 hours >in 10 % formalin and embed in OCT, but I have to cut with T -28 C in >microtome and mount on cool slides , otherwise I am not able to obtain >good vessel' lumen and plaque scrolls up or falls down and still I >receive 30% of distorted slices. Any advices will be appreciate. >Galina Deyneko >CV Department >Novartis Cambridge MA >617-871-7613 > > > > >--------------------------------- >Do you Yahoo!? > Take Yahoo! Mail with you! Get it on your mobile phone. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) --------------------------------- Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. From contact <@t> excaliburpathology.com Wed Dec 8 17:10:18 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Cryoembedding media woes, some advice and not a bash session Message-ID: <20041208231018.26334.qmail@web50306.mail.yahoo.com> O Gayle, I agree about OCT being the best. I have tried all the new ones that say they are as good or better and always go back to my bottle of OCT. It just infiltrates better, no matter what the tissue. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From fmonson <@t> wcupa.edu Wed Dec 8 17:46:54 2004 From: fmonson <@t> wcupa.edu (Monson, Frederick ) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC queries Bowman's layer and Descemet's Membrane-Mnemonic thread Message-ID: IF 'mne' were the mnemonic for "mnemonic", then it would also show up as the mnemonic for: Mom's Network Exchange Micro- and Nano-Engineering Minist?rio dos Neg?cios Estrangeiros (La) Maison de la Nature et de l'Environnement Museo Nacional de Escultura MultiNational Enterprises (mne's) Medical Nutrition Education Mac Newbold Enterprises Mechanical Nuclear Engineering "Mining Engineering Minor (MNE)" - so popular!!! Even when it should be: MEM!! MultiNational Experiment Sil Code for: "NABA: a language of Chad" etc. as in the 3-letter words = 972: http://www.cis.upenn.edu/~davidj2/sc/3s.html which leaves 16,604 unique three-letter combinations for more 3-letter abbreviations. Since mnemonics here on planet Earth are into redundancy we can surmise that since there is no logic involved there should be only one return for the 3-letter mnemonic, "aaa". One gets an entire page of "AAA" before one gets to another that signifies, of all things, the Amateur Astronomers Association. It is clear that for ANY 3-letter mnemonic (and the length of a mnemonic is NOT restricted in law or practice), there is an infinite number of potential uses, we are in for an increasingly difficult mental test. When things get less than logical the term "code" is substituted for"mnemonic" Thus, I have added an example from a DOD DOC: --------------------------------------------------------------- Type Program Full name 1 TBCOMCMD Command Designator 1 TBCOMID Command Identification Code 1 TBCOMPAC Command Performing Activity Code 1 TBCOMRIC Command Routing Identifier Code 1 TBDPTPAC Depot Performing Activity Code 1 TBDPTRIC Depot Routing Identifier Code 2 TFAPACE Army Procurement Appropriations Computerized Entry 2 TFORSCOM Deficiency Reporting System Command Address 2 TGINFFX Inflation Factor Table 1 TGSHIPQT Quantity Per Unit Package Code 1 THPALTST Procurement Administrative Leadtime Standards Table 1 THREJCD Reject Codes 3 TICLIFTM Controlled Item Logic Table 3 TIFRTSEC FRET Security Table 1 TJAFSICC Valid Air Force Service Item Control Center Activity Code by FSC Table 1 TJCMDACT Validate DOD Activity Code 1 TJFSCAC Category A and C FSC's Table 1 TJFSCARC Table 122 Army AAC 1 TJFSCSCM Federal Supply Class/Supply Category of Material/Essentiality Code Correlation Table 3 TJFSNKEY FSN Keyed Inquiry Routing 1 TJFSCTBL Federal Stock Class Table 1 TJICPTBL Inventory Control Point Routing Identifier Code Table 1 TJIMPTBL Inventory Manager Process Code Table 3 TJMINIRT NSNMDR Mini-Inquiry Routing 1 TJMOERUL Valid MOE Rules for Air Force, Marine Corps, Navy, and Coast Guard ----------------------------------------------------------------------------------------- This group demonstrates how incomprehensible it can become when one begins with the letter "T", follow it with "F", "G", "H", "I" or "J" and then 5 or 6 others. The above coded mnemonics are obviously related to the 'expanded names", but with little examination one can conclude that these are only aliases of mnemonics. Their number is so large that it is impossible to remember any, so in their constructs they do not make any serious attempt to suggest their meaning. This is the extreme example of, "I don't care a whit!" in the world of mnemonic design, or the prevailing attitude late on a Friday afternoon in the lexicographer's lab. Remember, once one begins with mnemonics, one will next need a list! Cheers, Fred Monson Frederick C. Monson, PhD Light, Electron, X-Ray and Scanning Probe Imaging and Analysis Center (LEXSPIAC) Large Scientific Instrument Core West Chester University S. Church St. and W. Rosedale Ave. West Chester, PA, 19320 610-738-0437 fmonson@wcupa.edu URL: http://darwin.wcupa.edu/casi -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Wednesday, December 08, 2004 3:48 PM To: Marshall Terry Dr, Consultant Histopathologist Cc: histonet@lists.utsouthwestern.edu; Paula Pierce Subject: Re: [Histonet] IHC queries Bowman's layer and Descemet's Membrane "Marshall Terry Dr, Consultant Histopathologist" wrote: > (A mnemonic for the spelling of mnemonic would be useful too). It's mnemonic because it's to help with the mnemory. (Sometimes you hear it pronounced "pneumonic"; people who do that need a mnemonic to remember that there isn't a u in mnemonic.) -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From immrstambo <@t> hotmail.com Wed Dec 8 17:56:33 2004 From: immrstambo <@t> hotmail.com (Christine Tambasco) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] please include me on the list Message-ID: From gcallis <@t> montana.edu Wed Dec 8 18:16:04 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] Cryoembedding media comments continued In-Reply-To: <20041208231018.26334.qmail@web50306.mail.yahoo.com> References: <20041208231018.26334.qmail@web50306.mail.yahoo.com> Message-ID: <6.0.0.22.1.20041208170446.01b20d68@gemini.msu.montana.edu> Dear Paula, I did not say OCT was THE best, only that it WORKS best for our particular (hmmm peculiar?) murine tissue work. I am keeping an open mind and maybe there will be another that works for me someday. I know people from other parts of USA who prefer other brands of cryomedias for their murine work. At least you do what we do and keep trying other cryomedias. Also, I think mfr's should know what has happened in special situations so they are aware of my problems with their product outside of a clinical situation. If you ever find another cryomedia you like, let me know - I'm all for new things that work well. Happy holidays At 04:10 PM 12/8/2004, you wrote: >O Gayle, I agree about OCT being the best. I have tried all the new ones >that say they are as good or better and always go back to my bottle of >OCT. It just infiltrates better, no matter what the tissue. > > >Paula Pierce, HTL(ASCP)HT > >Excalibur Pathology, Inc. >631 N. Broadway >Moore, OK 73160 >405-759-3953 >contact@excaliburpathology.com >www.excaliburpathology.com >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Histologia <@t> aol.com Wed Dec 8 19:03:58 2004 From: Histologia <@t> aol.com (Histologia@aol.com) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] opening in Denver Message-ID: <2b.6809ecaf.2ee8fe7e@aol.com> For those who would like to relocate to the midwest (Denver) area please watch for the upcoming posting. We will be needing a full time HT. Thanks in advance. Marina From Kemlo.Rogerson <@t> elht.nhs.uk Thu Dec 9 02:57:06 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] QI study[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5BD@bhrv-nt-11.bhrv.nwest.nhs.uk> I think the way to do it is to find your current level of mislabelling, and then choose to reduce it by say 10% month. In my opinion there is no 'acceptable' level, but the point of audit is to reach excellence by continual improvement; that obviates the premise of an 'acceptable' level. In my opinion. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: 08 December 2004 20:27 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] QI study[Scanned] Just wondering if anyone out there has ever done a QI study on mislabelled slides? My QI coordinator and Lab Director want me to do this for 2005. I am trying to identify an acceptable threshhold. 100% would be great, but I am a realist. Any ideas? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Thu Dec 9 03:05:42 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] RE: IHC on brain cryosections... help!! Message-ID: <17f1be17b8e5.17b8e517f1be@amc.uva.nl> Hello Anna, Besides all the good suggestions to your problem here is another one. Since you are dealing with mouse tissue and you haven't said anything special about your secondary antibody I assume this was just an ordinary reagent. Usually these reagents are meant for application on human tissues and therefore adsorbed for human immunoglobulins, but not for mouse immunoglobulins. There might be a reaction of your secondary reagent with endogenous immunoglobulins in your tissue sections, causing the background staining as you described. If true, this background also occurs when leaving out your primary antibody from the protocol. You can abolish this background by mixing 10% normal mouse serum with your secondary reagent. You have to do this 30-60 min (room temp.) before actually applying it to the tissue section. Furthermore, you should reconsider the use of 3% hydrogen peroxide on a cryostat tissue section. If there are lots of erythrocytes around there will be an air-bubble formation that is damaging the tissue section. Instead, use 0.3% hydrogen peroxide + 0.1% sodium azide in PBS or TBS (20 min, RT). This is perhaps less efficient with respect to endogenous peroxidase in neutrophils but definitely more gentle to your tissue sections. Good luck! Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: c.m.vanderloos@amc.uva.nl ----- Original Message ----- >From "Anna Elisse Beaudin" Date Tue, 7 Dec 2004 15:28:46 -0500 (EST) To histonet@lists.utsouthwestern.edu Subject [Histonet] IHC on brain cryosections... help!! Hi all, I am having a TON of trouble with basic IHC on mouse brain cryostat sections! I have reviewed tons of protocols from the web and from diferent papers.. and I am constantly getting confused between protocols for fixed, paraffin-embedded sections, and those for cryosections. Here is an overview of the protocol I am currently using: -Collect cryosections on slides, leave at room temp 1-2 hours (as I'm collecting) - Quick-fix in buffered PFA (4%) for 10 minutes - Rinse 3-10' PBS - Block for endogenous peroxidase activity in 3% hydrogen peroxide in PBS - Rinse 3 * 10 min Tris-buffered saline - Block in 1%BSA, .3% triton X in TBS - in primary o/n at 4 C - Rinse 3 * 10 min TBS - in secondary 1 hr at room temp - Rinse 3* 10 min TBS -- avitin-biotin (vector elite kit) for half hour at room temp -- Rinse 2 * 10 min TBS, 1*10 min TB pH7.6 -- in DAB 2-3 minutes With this protocol, I get terrible background in my negative controls, and cannot identify any stain in my positive slides. The background I'm getting does not appear to be cellular at all -- there is just brown gunk everywhere. Can anyone point me in the right direction? Thank you so much in advance for your help! Best, Anna Beaudin Division of Nutritional Sciences Cornell University Ithaca, NY From BMolinari <@t> heart.thi.tmc.edu Thu Dec 9 06:03:55 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] H and E vs. Masson Trichrome in cardiac injury Message-ID: Hi Natalie, We use Massons Trichrome for all our cardiac fibrosis studiea, paraffin and frozen. I a 0.1% light green counterstain. I am not involved in the evaluation end of the studies but will try and get that information and let you know. Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lapointe, Nathalie,Ph.D. Sent: Wednesday, December 08, 2004 12:24 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] H and E vs. Masson Trichrome in cardiac injury > Hi, > > I found that website by hazard and I'm very impress! > > I have another questions for you: > > I'm currently exploring some hormones that may cause myocardial injury in > adrenalectomized rats. > The heart were stained with hematoxylin and eosin for light microscopic > analysis (x40). A scale from 0-4 was used to score the level of myocardial > injury. A score of 0 represented no damage. A score of 1 represented the > presence of myocytes demonstrating early necrotic changes such as nuclear > pyknosis or karylysis, and oesinophil staining of the cytoplasm associated > with the presence of scattered neutrophilic infiltrates. A score of 2 was > given when one clear area of necrosis (loss of myocardial cells with heavy > neutrophilic infiltrates) was observed. When two or more separate areas of > necrosis were found (implicating the presence of two different myocardial > infarctions in the same heart), but the areas localized and compromised less > then 50% of the ventricular wall, the hearts received a score of 3. A score of > 4 was assigned to hearts that demonstrated extensive areas of necrosis > compromising more than 50% of wither the left or the right ventricle. > > I'm looking for another way then the above score to measure cardiac injury. > Based to the fact that Hematoxylin is a dark purplish dye that will stain the > chromatin within the nucleus (therefore also monocyte infiltration), and that > Eosin is an orangish pink to red dye that stains the cytoplasmic material > including connective tissue and collagen (therefore doesn't stain necrotic > tissues), I was wondering if a software in which I could measure the contrast > of color related to injury vs. intact tissue exist? Does this same software > (if available) can also calculated the number of dark purplish dye (therefore > level of inflammation)? > > We also stained the same heart with Masson Trichrome. Do you think that > measuring the % of blue staining (collagen) in the whole heart at a > magnification of X4 is equivalent to H and E score described above? How can I > know if the blue staining with Masson Trichrome is fibrosis rather then > collagen or only red Sirus staining can measure fibrosis? > Thank you very much for all your help. Best, > Nathalie Lapointe > Brigham and Women's Hospital > LMRC #214 > 221 Longwood Ave > Boston, MA, USA > 02115 > (617) 515-5936 > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Thu Dec 9 06:34:02 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] QI study Message-ID: <83AACDB0810528418AA106F9AE9B7F7E50766D@sjhaexc02.sjha.org> I think 100% is what we should achieve, but as long as we are human we are bound to have errors. Documenting and making the staff aware of any errors helps to reduce them, and the goal to achieve 100% is one to always be working toward. Graphs are a good way to show progress. My 2 cents worth...j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Angela Bitting Sent: Wednesday, December 08, 2004 3:27 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] QI study Just wondering if anyone out there has ever done a QI study on mislabelled slides? My QI coordinator and Lab Director want me to do this for 2005. I am trying to identify an acceptable threshhold. 100% would be great, but I am a realist. Any ideas? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From NSEARCY <@t> swmail.sw.org Thu Dec 9 06:35:30 2004 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] University Of Miami Contact Message-ID: Anyone have a telephone contact with histology person @ the University of Miami? Pathologist wants tech to contact microwave precessing expert. Thanks From JWEEMS <@t> sjha.org Thu Dec 9 06:39:34 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] University Of Miami Contact Message-ID: <83AACDB0810528418AA106F9AE9B7F7E50766E@sjhaexc02.sjha.org> That would be Dr. Morales @ 305-585-6103. You might want to contact your Sakura rep who can set up a site visit. Dr. Morales has scheduled days that he does the site visits so that he can give his full time to the visitor. Cheers! j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Nita Searcy Sent: Thursday, December 09, 2004 7:36 AM To: histonet@lists.utsouthwestern.edu Cc: Mae Lopez Subject: [Histonet] University Of Miami Contact Anyone have a telephone contact with histology person @ the University of Miami? Pathologist wants tech to contact microwave precessing expert. Thanks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From lpjones <@t> srhs-pa.org Thu Dec 9 10:03:38 2004 From: lpjones <@t> srhs-pa.org (Jones, Laura) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC ~ Tissue Falling Off Slides Message-ID: <8E7AD740937B954F947F0DB4467EFEE056E93D@mail.srhs-pa.org> Hello fellow Histonetters. We have suddenly started to have problems with tissue falling off slides during IHC processing. We suspect the problem may be our slides, although we have been using the same ones from Mercedes Medical for a while and they have worked beautifully! We have contacted them, and they are investigating and very kindly replacing our stock, but in the meantime, our Pathologist has asked me to ask all of you where else we can find 25 X 75 silinated slides. We use the Zymed ST-5050 automated stainer, and the 1 X 3 slides are sometimes a bit to big to fit on the carousel. Just for additional info, we seem to be experiencing problems mostly with the more alkaline retreival solutions - AR-10 (Biogenex) and EDTA (Zymed) and Trilogy (Cell Marque). We use the Black & Decker steamer, and have adjusted our times in retrieval from 75 minutes (!) to 20 minutes, per Pathologist direction. We have also tried 20 minutes heating, then adding the slides for 20 minutes of boiling, then allowing them to stand for 20-30 minutes. Thanks in advance for all of your knowledge! The Histochicks at Sharon Regional From JGordon <@t> cellmarque.com Thu Dec 9 10:14:42 2004 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC ~ Tissue Falling Off Slides Message-ID: Laura, what drying technique are you using? While the problem may possibly be the slides, often we are able to solve tissue adhesion problems with proper slide drying. We use standard positively charged slides with no special coating and no special adhesive in the water bath. We pretreat slides in an electric pressure cooker for 15 minutes with Trilogy EDTA, which is a pretty aggressive pretreatment, and we never have recurring problems with tissue adhesion. Our slide drying technique is 2 hours at 60 degrees Centigrade for most tissues (super sensitive tissues like fatty breast we will dry for 4 hours at 60 degrees), and the tissue stays on just fine in the pressure cooker pretreatment. Jeff Gordon Cell Marque Corp. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jones, Laura Sent: Thursday, December 09, 2004 10:04 AM To: Histonet (E-mail) Subject: [Histonet] IHC ~ Tissue Falling Off Slides Hello fellow Histonetters. We have suddenly started to have problems with tissue falling off slides during IHC processing. We suspect the problem may be our slides, although we have been using the same ones from Mercedes Medical for a while and they have worked beautifully! We have contacted them, and they are investigating and very kindly replacing our stock, but in the meantime, our Pathologist has asked me to ask all of you where else we can find 25 X 75 silinated slides. We use the Zymed ST-5050 automated stainer, and the 1 X 3 slides are sometimes a bit to big to fit on the carousel. Just for additional info, we seem to be experiencing problems mostly with the more alkaline retreival solutions - AR-10 (Biogenex) and EDTA (Zymed) and Trilogy (Cell Marque). We use the Black & Decker steamer, and have adjusted our times in retrieval from 75 minutes (!) to 20 minutes, per Pathologist direction. We have also tried 20 minutes heating, then adding the slides for 20 minutes of boiling, then allowing them to stand for 20-30 minutes. Thanks in advance for all of your knowledge! The Histochicks at Sharon Regional _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpjones <@t> srhs-pa.org Thu Dec 9 10:43:57 2004 From: lpjones <@t> srhs-pa.org (Jones, Laura) Date: Fri Sep 16 15:24:23 2005 Subject: [Histonet] IHC ~ Tissue Falling Off Slides, drying procedure.slides Message-ID: <8E7AD740937B954F947F0DB4467EFEE056E93E@mail.srhs-pa.org> Hey Jeff! We have tried many variations on drying as well. Generally, we cut the tissue, let it stand and drain/air dry at room temp for 0.5-1.0 hour, then place on the slide dryer at 65-70 for another hour, then place in the slide oven at 80 to melt - usually about 20 minutes. We have also tried microwave drying. We have also dipped the slides (after the tissue is on them) into Poly-L-Lysine. Even slides that are cut on Saturday, dry all weekend and not retrieved until Monday morning are troublesome. Dottie - we happen to have a few boxes of Superfrost Plus slides here from our Cytology department and are cutting some test slides as I type this. Thanks for the suggestion! From gcallis <@t> montana.edu Thu Dec 9 11:09:17 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] IHC ~ Tissue Falling Off Slides, drying procedure.slides In-Reply-To: <8E7AD740937B954F947F0DB4467EFEE056E93E@mail.srhs-pa.org> References: <8E7AD740937B954F947F0DB4467EFEE056E93E@mail.srhs-pa.org> Message-ID: <6.0.0.22.1.20041209100238.01b2b650@gemini.msu.montana.edu> Laura, Do you use tap water or distilled water in a waterbath? Erie Plus Charge slide box insert instructions for use recommends this to avoid any hard water components. Newcomer Supply has some super slides that may be a possibility along with the Erie Superfrost Plus slides. Marsha , at Newcomer, maintains that the tissue portion of paraffin section must come into very quick contact with a positive charge slide surface during picking up process. If you have questions, she would be glad to discuss it with you and she will send sample of her slides to try. They do have a website. There are also low temperature methods for retrieval using 99C waterbaths. At 09:43 AM 12/9/2004, you wrote: >Hey Jeff! We have tried many variations on drying as well. Generally, we >cut the tissue, let it stand and drain/air dry at room temp for 0.5-1.0 >hour, then place on the slide dryer at 65-70 for another hour, then place in >the slide oven at 80 to melt - usually about 20 minutes. > >We have also tried microwave drying. > >We have also dipped the slides (after the tissue is on them) into >Poly-L-Lysine. > >Even slides that are cut on Saturday, dry all weekend and not retrieved >until Monday morning are troublesome. > > >Dottie - we happen to have a few boxes of Superfrost Plus slides here from >our Cytology department and are cutting some test slides as I type this. >Thanks for the suggestion! > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Johnson.Sandra4 <@t> mayo.edu Thu Dec 9 11:21:39 2004 From: Johnson.Sandra4 <@t> mayo.edu (Johnson, Sandra J. (MCS)) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Sakura Express Microwave Processor Message-ID: <58FEAD34B9A42C4A949BAB704DB8DA101A52A7@excsrv69.mayo.edu> Hello out there: My supervisors have asked whether anyone out there is using the Express Microwave Processor. We have only received information from beta testing sites and would like some users' opinions prior to purchasing. Thanks, Sandy Johnson, HTL IHC Lead Tech - Histology Mayo Clinic Scottsdale From glv4708 <@t> bjc.org Thu Dec 9 11:54:24 2004 From: glv4708 <@t> bjc.org (Gretchen Vollmer) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] HPV ISH Control Slides Message-ID: We have just started using our Ventana stainer for HPV ISH (high risk only). I have been encouraged to make my own positive control slides to reduce costs. Has anyone done this with success? If so, would you share your methodology, if different from Ventana's protocol? Any advise would be appreciated. Thanks. From cwscouten <@t> myneurolab.com Thu Dec 9 12:00:13 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] RE: Cryo Mbed Message-ID: Yes, Cryo M Bed is firmer and more supportive. Try to inject some of whatever you use into the lumen before freezing. That would help greatly. Call me off histonet and we will see if we can arrange a sample. Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com ________________________________ From: Galina Deyneko [mailto:galinadeyneko@yahoo.com] Sent: Wednesday, December 08, 2004 12:37 PM To: histonet@lists.utsouthwestern.edu Cc: Charles Scouten Subject: Cryo Mbed Hi Dr.Scouten and histonetters, Does it mean that Cryo M Bed more "supportive" than OCT. I work with murine carotid arteries with atheroscl. plaque. .I fix them for 2 hours in 10 % formalin and embed in OCT, but I have to cut with T -28 C in microtome and mount on cool slides , otherwise I am not able to obtain good vessel' lumen and plaque scrolls up or falls down and still I receive 30% of distorted slices. Any advices will be appreciate. Galina Deyneko CV Department Novartis Cambridge MA 617-871-7613 ________________________________ Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. From KarBieber <@t> aol.com Thu Dec 9 13:09:32 2004 From: KarBieber <@t> aol.com (KarBieber@aol.com) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Microwave staining Message-ID: <8b.1c261a8e.2ee9fcec@aol.com> I'd like to know what kind of microwaves other labs are using to perform special stains. We've gone through a series of regular household brands but they keep burning out. Any suggestions? Karen From cwscouten <@t> myneurolab.com Thu Dec 9 13:37:36 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Microwave staining Message-ID: Try a Pelco Neurowave. See Link. It is the only FDA registered histology microwave. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=472001&catdesc=Histology+Equipment&CatThreeID=714&CatOneID=4&subcatdesc=Microwave+Reaction+Facilitation&idsubcategory=204 Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of KarBieber@aol.com Sent: Thursday, December 09, 2004 1:10 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave staining I'd like to know what kind of microwaves other labs are using to perform special stains. We've gone through a series of regular household brands but they keep burning out. Any suggestions? Karen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pzeitlow <@t> bbpllab.com Thu Dec 9 14:02:48 2004 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] ventana xt upc requirements Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A0BEB1C@bbplsrv1.bbpl> Can anyone share what back up power they are using successfully with the XT? I am interested in having power at least 30 minutes for two instruments. Pat Z Department of Molecular Pathology Boyce and Bynum Pathology Labs, P.C. From BennettW <@t> pac.dfo-mpo.gc.ca Thu Dec 9 15:44:27 2004 From: BennettW <@t> pac.dfo-mpo.gc.ca (BennettW@pac.dfo-mpo.gc.ca) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Leica/Jung Histoembedder Message-ID: <7CBBD627E4E688499349A5D11D078316020DBB2B@msgpacpbs.rhq.pac.dfo-mpo.gc.ca> Hello All, Does anyone have this type of embedding centre? Mine is displaying ERROR:#07 on the display screen and I have tried everything to get rid of it. It seems to work fine most of the time although sometimes it won't dispense wax and I have to turn off the power and then back on to make it work again. The local distributor is reluctant to try and diagnose this over the phone and wants to send a technician at $250.00/hr plus travel time. As the local distributor is a 2 hr. boat ride away, I thought that someone in the group could help me out, please, please. Cheers Bill Bennett Histologist Fisheries and Oceans Canada Nanaimo, B.C. Canada From mari.ann.mailhiot <@t> leica-microsystems.com Thu Dec 9 16:40:17 2004 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Leica/Jung Histoembedder Message-ID: Hi Bill Please contact me here at Leica and we can discuss. The error is in reference to the mold tray. Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com BennettW@pac.dfo-mpo.gc.ca Sent by: To: histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] Leica/Jung Histoembedder 12/09/2004 03:44 PM Hello All, Does anyone have this type of embedding centre? Mine is displaying ERROR:#07 on the display screen and I have tried everything to get rid of it. It seems to work fine most of the time although sometimes it won't dispense wax and I have to turn off the power and then back on to make it work again. The local distributor is reluctant to try and diagnose this over the phone and wants to send a technician at $250.00/hr plus travel time. As the local distributor is a 2 hr. boat ride away, I thought that someone in the group could help me out, please, please. Cheers Bill Bennett Histologist Fisheries and Oceans Canada Nanaimo, B.C. Canada _______________________________________________ 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 JGordon <@t> cellmarque.com Thu Dec 9 22:53:36 2004 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] IHC ~ Tissue Falling Off Slides, drying procedure.slides Message-ID: Laura, here are the issues that we deal with most in regards to tissue adhesion: 1) Drying (most common). When you room temperature dry the slides, the water will dry from the outside in. Once the water is out from the edge of the slide, the paraffin sort of adheres itself to the slide around the tissue, creating a seal that doesn't allow any more water to escape from under the paraffin. It can sit there for a month and still have water trapped under the tissue in this condition. When you put the slides in the oven, the paraffin melts allowing water to escape from under the seal and under the tissue, thereby allowing the tissue to fully adhere to the slide without any water barrier separating any part of it. Since you are already drying for 2+ hours, I would recommend just putting them in the 60 degree oven for that entire time. If you dry in the microwave, we usually dry for a total of 3 minutes under power. We run it for a minute and a half at full power, then let it set for 30 seconds to a minute (enough to allow the paraffin to solidify again) and then run it for another minute and a half and let it cool again. 2) Retrieval pH. The higher pH solutions are harsher on the tissue than other solutions, mainly because raising the pH will literally eat the tissue up (think about what Drano would do to your skin, but on a much lesser scale). That is where we usually promote Trilogy. It usually isn't as harsh as high pH (9 or 10) or straight EDTA because of the emulsifiers in it that help bring the pH to a more tolerable level for the tissue. Many labs that have used Trilogy brought it on board because of the harshness of other solutions that they were buying or making themselves. The more basic that you make the solution, the more likely it is that you will have tissue/morphology problems, regardless of the slide preparation. 3) Waterbath contaminates. Anything in the water can affect the adhesion of the tissue. Deionized water is preferred. 4) Tissue thickness. We never recommend that tissues cut at more than 4 microns be used for clinical IHC. We cut ours at 3-4 microns standard regardless of the tissue. If you cut them thick they are more likely to lose adhesion and can give background, as well. 5) Slide handling. Don't touch the flat surface of the slides or you can draw the charge off of them. Hold them by the label area. 6) Formalin pH. This is often taken for granted, but can have a major affect on tissue adhesion. If the formalin has too many hydrogen ions (is too acidic), it will pass a positive charge onto the tissue that it is fixing. Not good. The negative charge of the skin is what makes it adhere to the positive charge of the histology slide. If the tissue carries positive charge due to acidic formalin, and the slide is positively charged, then you know what like charges do. Try to keep the formalin as close to neutral pH as possible. 7) It is possible that a certain lot of slides that you receive could be bad. It happens to everyone sometime. 8) Retrieval time. We recommend using a pressure cooker, and one major reason is the speed. There are different suggested protocols for heat retrieving in a pressure cooker, and different pressure cookers on the market. Regardless of the suggested protocol or the brand of pressure cooker that you use, pressure cookers have one thing in common: they are faster than steamer methods. Pressure cookers reach a higher heat, and they are more aggressive than other pretreatments, but you have the slides in the hot solution for a much shorter time, which can be key. YOu even recognize that from reducing your steamer retrieval time from 75 minutes to 20 minutes in order to try and improve the adhesion. By using a pressure cooker (whether it is from us or from a competitor or from a kitchen store) you can improve your stains, increase your turnaround time, and also possibly help your tissue adhesion due to the short period that the slides are in hot solution (with Trilogy we don't require a cooldown period after heat retrieval). This is NOT a pitch for our pressure cooker. We don't care what pressure cooker you use, but we do recommend using the pressure cooker for your heat retrieval. I don't expect any of this information be news to anyone on here. You all have been in this industry at least twice as long as I have been, but I try to help troubleshoot often and these are the factors that play into tissue adhesion questions that I try to help workout for clients. These are just reminders of things to check if you have an adhesion problem. I hope that this helps. Jeff Gordon Cell Marque Corp. 1-800-665-7284, Ext. 12 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jones, Laura Sent: Thursday, December 09, 2004 10:44 AM To: Histonet (E-mail) Subject: [Histonet] IHC ~ Tissue Falling Off Slides, drying procedure.slides Hey Jeff! We have tried many variations on drying as well. Generally, we cut the tissue, let it stand and drain/air dry at room temp for 0.5-1.0 hour, then place on the slide dryer at 65-70 for another hour, then place in the slide oven at 80 to melt - usually about 20 minutes. We have also tried microwave drying. We have also dipped the slides (after the tissue is on them) into Poly-L-Lysine. Even slides that are cut on Saturday, dry all weekend and not retrieved until Monday morning are troublesome. Dottie - we happen to have a few boxes of Superfrost Plus slides here from our Cytology department and are cutting some test slides as I type this. Thanks for the suggestion! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMacDonald <@t> mtsac.edu Fri Dec 10 02:58:16 2004 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] iron stain and carbol fuchsin Message-ID: A blood film was stained with carbol fuchsin. It was intend an iron stain. The smear was decolorized quickly (few quick in acid alcohol. Does anyone know the affect this would have on iron stain? Can it be saved? Thanks in Jennifer MacDonald From spoulos <@t> saa.ars.usda.gov Fri Dec 10 07:33:57 2004 From: spoulos <@t> saa.ars.usda.gov (Sylvia Poulos) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] NADH-TR stain Message-ID: Hi everyone, I just finished staining some skeletal muscle sections with NADH-tetrazolium reductase. It worked great, as usual, but I see some fibers that are well stained around the outer perimeter with very little staining in the center of the fiber. I don't think it's artifact but wanted to ask people with more experience... Is there any clinical significance to this? Is there anything else I should stain with? I've done Oil Red O and the pattern is similar. As always, thanks for the help. Sylvia P. Poulos USDA-ARS-Animal Physiology Research Unit Athens, GA 30605 706-542-4694 706-542-0399 (fax) From SCheasty <@t> memorialcare.org Fri Dec 10 07:39:44 2004 From: SCheasty <@t> memorialcare.org (Sandy Cheasty) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] NCCLS Microwave Device Use Proposed Guideline Message-ID: <59FE4C55358C6A42B84BAEC6092CEDBC05ACF53E@sbnt7> I have the GP28-P guideline and read it but am unsure about its recommendations or limitations in using a microwave for special stains only. Is there any consensus among histology labs as to its interpretation? Is it still OK to use a commercial microwave from Wal-Mart without special exhaust system as long as you don't microwave formaldehyde, osmium tetroxide, lead acetate etc. (Paragraph 4.4) and do annual leakage tests? Thanks! Elvis ______________________________________________________________________________ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm From David.Edmondson <@t> christie-tr.nwest.nhs.uk Fri Dec 10 08:31:57 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Klane-Staff embedding centre Message-ID: Hello, Is there anyone out there with knowlage of a transformer unit/PSU for one of these embedding centres, that is to say an spare one, that we could aquire? We are looking for backup and Bill indicated that they were manufactured to order and so unavailable now. David Edmondson Christie Hospital Manchester UK From DonnaWillis <@t> texashealth.org Fri Dec 10 09:07:48 2004 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] NCCLS Microwave Device Use Proposed Guideline Message-ID: It is against OSHA guideline (OSHA 29 CFR 1910.303(b)(2))to use a commercial microwave (exp. Wal-Mart). The warranty of commercial units state that the instrument is not to be used for anything except its original use, food preparation. The OSHA guideline would include any equipment in the lab. If someone gets injured the lab would be at fault not the manufacture. Donna Willis Histology Lab Manager Harris Methodist Fort Worth,Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sandy Cheasty Sent: Friday, December 10, 2004 7:40 AM To: histonet@pathology.swmed.edu Subject: [Histonet] NCCLS Microwave Device Use Proposed Guideline I have the GP28-P guideline and read it but am unsure about its recommendations or limitations in using a microwave for special stains only. Is there any consensus among histology labs as to its interpretation? Is it still OK to use a commercial microwave from Wal-Mart without special exhaust system as long as you don't microwave formaldehyde, osmium tetroxide, lead acetate etc. (Paragraph 4.4) and do annual leakage tests? Thanks! Elvis ____________________________________________________________________________ __ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From jkiernan <@t> uwo.ca Fri Dec 10 09:39:34 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] iron stain and carbol fuchsin References: Message-ID: <41B9C336.38F79AF3@uwo.ca> Acid generally removes iron, so probably not. John Kiernan London, Canada. ______________________________________________ Jennifer MacDonald wrote: > > A blood film was stained with carbol fuchsin. It was intend= ed to be > an iron stain. The smear was decolorized quickly (few quick = dips) > in acid alcohol. Does anyone know the affect this would have on= a > iron stain? Can it be saved? ______________________________________________ From kmerriam2003 <@t> yahoo.com Fri Dec 10 09:54:55 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] IHC ~ Tissue Falling Off Slides, drying procedure.slides In-Reply-To: Message-ID: <20041210155455.89502.qmail@web52505.mail.yahoo.com> Laura, What type of tissue are you using? Are you using superfrost plus or other charged slides? I have been told that these have a shelf-life and after a certain amount of time (~ 18 months or so), the charge goes away. This might be something to look into. Kim Merriam Novartis Cambridge, MA Jeff Gordon wrote: Laura, here are the issues that we deal with most in regards to tissue adhesion: 1) Drying (most common). When you room temperature dry the slides, the water will dry from the outside in. Once the water is out from the edge of the slide, the paraffin sort of adheres itself to the slide around the tissue, creating a seal that doesn't allow any more water to escape from under the paraffin. It can sit there for a month and still have water trapped under the tissue in this condition. When you put the slides in the oven, the paraffin melts allowing water to escape from under the seal and under the tissue, thereby allowing the tissue to fully adhere to the slide without any water barrier separating any part of it. Since you are already drying for 2+ hours, I would recommend just putting them in the 60 degree oven for that entire time. If you dry in the microwave, we usually dry for a total of 3 minutes under power. We run it for a minute and a half at full power, then let it set for 30 seconds to a minute (enough to allow the paraffin to solidify again) and then run it for another minute and a half and let it cool again. 2) Retrieval pH. The higher pH solutions are harsher on the tissue than other solutions, mainly because raising the pH will literally eat the tissue up (think about what Drano would do to your skin, but on a much lesser scale). That is where we usually promote Trilogy. It usually isn't as harsh as high pH (9 or 10) or straight EDTA because of the emulsifiers in it that help bring the pH to a more tolerable level for the tissue. Many labs that have used Trilogy brought it on board because of the harshness of other solutions that they were buying or making themselves. The more basic that you make the solution, the more likely it is that you will have tissue/morphology problems, regardless of the slide preparation. 3) Waterbath contaminates. Anything in the water can affect the adhesion of the tissue. Deionized water is preferred. 4) Tissue thickness. We never recommend that tissues cut at more than 4 microns be used for clinical IHC. We cut ours at 3-4 microns standard regardless of the tissue. If you cut them thick they are more likely to lose adhesion and can give background, as well. 5) Slide handling. Don't touch the flat surface of the slides or you can draw the charge off of them. Hold them by the label area. 6) Formalin pH. This is often taken for granted, but can have a major affect on tissue adhesion. If the formalin has too many hydrogen ions (is too acidic), it will pass a positive charge onto the tissue that it is fixing. Not good. The negative charge of the skin is what makes it adhere to the positive charge of the histology slide. If the tissue carries positive charge due to acidic formalin, and the slide is positively charged, then you know what like charges do. Try to keep the formalin as close to neutral pH as possible. 7) It is possible that a certain lot of slides that you receive could be bad. It happens to everyone sometime. 8) Retrieval time. We recommend using a pressure cooker, and one major reason is the speed. There are different suggested protocols for heat retrieving in a pressure cooker, and different pressure cookers on the market. Regardless of the suggested protocol or the brand of pressure cooker that you use, pressure cookers have one thing in common: they are faster than steamer methods. Pressure cookers reach a higher heat, and they are more aggressive than other pretreatments, but you have the slides in the hot solution for a much shorter time, which can be key. YOu even recognize that from reducing your steamer retrieval time from 75 minutes to 20 minutes in order to try and improve the adhesion. By using a pressure cooker (whether it is from us or from a competitor or from a kitchen store) you can improve your stains, increase your turnaround time, and also possibly help your tissue adhesion due to the short period that the slides are in hot solution (with Trilogy we don't require a cooldown period after heat retrieval). This is NOT a pitch for our pressure cooker. We don't care what pressure cooker you use, but we do recommend using the pressure cooker for your heat retrieval. I don't expect any of this information be news to anyone on here. You all have been in this industry at least twice as long as I have been, but I try to help troubleshoot often and these are the factors that play into tissue adhesion questions that I try to help workout for clients. These are just reminders of things to check if you have an adhesion problem. I hope that this helps. Jeff Gordon Cell Marque Corp. 1-800-665-7284, Ext. 12 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jones, Laura Sent: Thursday, December 09, 2004 10:44 AM To: Histonet (E-mail) Subject: [Histonet] IHC ~ Tissue Falling Off Slides, drying procedure.slides Hey Jeff! We have tried many variations on drying as well. Generally, we cut the tissue, let it stand and drain/air dry at room temp for 0.5-1.0 hour, then place on the slide dryer at 65-70 for another hour, then place in the slide oven at 80 to melt - usually about 20 minutes. We have also tried microwave drying. We have also dipped the slides (after the tissue is on them) into Poly-L-Lysine. Even slides that are cut on Saturday, dry all weekend and not retrieved until Monday morning are troublesome. Dottie - we happen to have a few boxes of Superfrost Plus slides here from our Cytology department and are cutting some test slides as I type this. Thanks for the suggestion! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? The all-new My Yahoo! – What will yours do? From m-degutes <@t> northwestern.edu Fri Dec 10 10:31:24 2004 From: m-degutes <@t> northwestern.edu (Mathew DeGutes) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Quenching HRP in double IHC Message-ID: <200412101631.iBAGVekF014581@casbah.it.northwestern.edu> Hi, I am a Research Tech at Northwestern University and I have run into a problem that to me seems completely bizarre. I am performing double fluorescent IHC stains using HRP for both stains and using cy3 and fluorescein to develop on frozen sections. My problem is this. After my first development with cy3 I try to quench the HRP using 6% H2O2 for 10 min. What then happens is when I go to develop my 2nd stain it is being developed by the HRP from the first stain that was apparently never quenched at all. I have tested this directly and it continues to be true. 6% H2O2 should easily knock the HRP out, but it isn't. All my conventional wisdom is sort of out the window. The only thing that I could think would be the cause is water. Does water destroy H2O2? Could that inhibit H2O2 action is PBS necessary to preserve it? Any help is appreciated. From JWEEMS <@t> sjha.org Fri Dec 10 11:38:18 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] iron stain and carbol fuchsin Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507699@sjhaexc02.sjha.org> That's why iron doesn't work well on the decaled biopsy... Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of John Kiernan Sent: Friday, December 10, 2004 10:40 AM To: Jennifer MacDonald Cc: histonet@pathology.swmed.edu Subject: Re: [Histonet] iron stain and carbol fuchsin Acid generally removes iron, so probably not. John Kiernan London, Canada. ______________________________________________ Jennifer MacDonald wrote: > > A blood film was stained with carbol fuchsin. It was intend= ed to be > an iron stain. The smear was decolorized quickly (few quick = dips) > in acid alcohol. Does anyone know the affect this would have on= a > iron stain? Can it be saved? ______________________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From gcallis <@t> montana.edu Fri Dec 10 11:54:05 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Postive charge slides, section loss and drying Message-ID: <6.0.0.22.1.20041210105228.01b17ad0@gemini.msu.montana.edu> > >Jeff, > >It was pointed out to us (by Marcia (sp?) from Newcomer Supply) - it is >important HOW a section is picked up. One should make sure the tissue >portion of paraffin section comes into contact with positive charged >surface very quickly during picking up motion. This permits the tissue to >adhere better, without so much water underneath it, onto the positive >charged surface. IF the section is in good contact with the slide, there >will be better interaction of plus charge to tissue, and you would not >have to depend so much on paraffin melting to allow water to escape under >this + / - seal. >Vertical draining os sllides in a polypropylene slide holder is important >before loading slides in horizonal position in a rack. We keep a razor >blade handy to release water blebs at section edge before racking up and >drying. We have actually dried sections on Plus charge slides for 1 hour >at 60C and still had water present under paraffin. This ceased after >tweaking picking up technic and vertical drying. > >At 09:53 PM 12/9/2004, you wrote: > >>1) Drying (most common). When you room temperature dry the slides, the >>water will dry from the outside in. Once the water is out from the edge >>of the slide, the paraffin sort of adheres itself to the slide around the >>tissue, creating a seal that doesn't allow any more water to escape from >>under the paraffin. It can sit there for a month and still have water >>trapped under the tissue in this condition. When you put the slides in >>the oven, the paraffin melts allowing water to escape from under the seal >>and under the tissue, thereby allowing the tissue to fully adhere to the >>slide without any water barrier separating any part of it. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From tpmorken <@t> labvision.com Fri Dec 10 12:08:34 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Postive charge slides, section loss and drying Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA257F790@usca0082k08.labvision.apogent.com> Ah, the lore of histology " When you room temperature dry the slides, the water will dry from the outside in. Once the water is out from the edge of the slide, the paraffin sort of adheres itself to the slide around the tissue, creating a seal that doesn't allow any more water to escape from under the paraffin." " We keep a razor blade handy to release water blebs at section edge before racking up and drying." "Vertical draining of slides in a polypropylene slide holder is important before loading slides in horizonal position in a rack." I'll add my technique (such as it is) to the lore of histology: To lessen water entrapment I don't let the slide just passively drain. After picking up a section I do a quick wicking of exess water by holding the slide vertically on top of a paper towel and then give the slide a quick " wrist snap" to force out water from under the section (Think of snapping a whip - it's all in the wrist action!). I'm careful about fragile tissue and haven't had any problems with tissue damage doing that. It seems to get rid of most of the water under the section. If the section is especially fragile I just do the wicking, but do it on the short and long edges of the slide. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Friday, December 10, 2004 9:54 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Postive charge slides, section loss and drying > >Jeff, > >It was pointed out to us (by Marcia (sp?) from Newcomer Supply) - it >is >important HOW a section is picked up. One should make sure the tissue >portion of paraffin section comes into contact with positive charged >surface very quickly during picking up motion. This permits the tissue to >adhere better, without so much water underneath it, onto the positive >charged surface. IF the section is in good contact with the slide, there >will be better interaction of plus charge to tissue, and you would not >have to depend so much on paraffin melting to allow water to escape under >this + / - seal. >Vertical draining os sllides in a polypropylene slide holder is important >before loading slides in horizonal position in a rack. We keep a razor >blade handy to release water blebs at section edge before racking up and >drying. We have actually dried sections on Plus charge slides for 1 hour >at 60C and still had water present under paraffin. This ceased after >tweaking picking up technic and vertical drying. > >At 09:53 PM 12/9/2004, you wrote: > >>1) Drying (most common). When you room temperature dry the slides, >>the >>water will dry from the outside in. Once the water is out from the edge >>of the slide, the paraffin sort of adheres itself to the slide around the >>tissue, creating a seal that doesn't allow any more water to escape from >>under the paraffin. It can sit there for a month and still have water >>trapped under the tissue in this condition. When you put the slides in >>the oven, the paraffin melts allowing water to escape from under the seal >>and under the tissue, thereby allowing the tissue to fully adhere to the >>slide without any water barrier separating any part of it. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From denise <@t> pclv.net Fri Dec 10 12:56:48 2004 From: denise <@t> pclv.net (Denise) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Employment opportunity Seattle/Everett Area Message-ID: Phoenix Central Laboratory for Veterinarians, Inc. has a position available for a Histology Technician with ASCP certification (or eligibility). Phoenix Central Laboratory, (PCLV), is a veterinary reference laboratory that provides diagnostic services to more than 800 clinics located throughout the Pacific Northwest and Alaska. We are dedicated to providing rapid, quality service. Our laboratory is located in Everett, Washington, just thirty minutes north of downtown Seattle. Visit our web site at www.pclv.net. PCLV offers a competitive salary and benefit package including health and dental coverage, personal time, and a 401K plan. Please submit your resume to Denise DeRouchey, c/o Phoenix Central Laboratory, 11620 Airport Road, Everett, WA 98204. Alternately, you may fax your resume to 425-355-3676. From SCheasty <@t> memorialcare.org Fri Dec 10 13:58:19 2004 From: SCheasty <@t> memorialcare.org (Sandy Cheasty) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Microwave Devices and Proposed Guidelines Message-ID: <59FE4C55358C6A42B84BAEC6092CEDBC05ACF53F@sbnt7> I know there are thousands of histology labs that use commercial microwaves safely and have used them safely for years. Does everyone: -violate OSHA? -lie to their lab managers? There must be some way of continuing to use commercial microwave ovens for special stains without being subversive. Don't get me wrong, I usually enjoy being subversive. Can someone please give me a justification for using a commercial microwave or tell me where I can get an OSHA approved one for less than $1000? Thanks again, Elvis -----Original Message----- From: Willis, Donna [mailto:DonnaWillis@texashealth.org] Sent: Friday, December 10, 2004 7:08 AM To: Sandy Cheasty; histonet@pathology.swmed.edu Subject: RE: [Histonet] NCCLS Microwave Device Use Proposed Guideline It is against OSHA guideline (OSHA 29 CFR 1910.303(b)(2))to use a commercial microwave (exp. Wal-Mart). The warranty of commercial units state that the instrument is not to be used for anything except its original use, food preparation. The OSHA guideline would include any equipment in the lab. If someone gets injured the lab would be at fault not the manufacture. Donna Willis Histology Lab Manager Harris Methodist Fort Worth,Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sandy Cheasty Sent: Friday, December 10, 2004 7:40 AM To: histonet@pathology.swmed.edu Subject: [Histonet] NCCLS Microwave Device Use Proposed Guideline I have the GP28-P guideline and read it but am unsure about its recommendations or limitations in using a microwave for special stains only. Is there any consensus among histology labs as to its interpretation? Is it still OK to use a commercial microwave from Wal-Mart without special exhaust system as long as you don't microwave formaldehyde, osmium tetroxide, lead acetate etc. (Paragraph 4.4) and do annual leakage tests? Thanks! Elvis ____________________________________________________________________________ __ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. ______________________________________________________________________________ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm From Luis.Chiriboga <@t> med.nyu.edu Fri Dec 10 16:12:07 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Microtomy Message-ID: Hi everyone Does anyone know of a text that describes (in detail) the physics and geometry of tissue sectioning? I'm looking for all the gory details....... Thanks Luis From MadaryJ <@t> MedImmune.com Fri Dec 10 16:21:56 2004 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Validated Accessioning Program, Freezer Maintenance Message-ID: <83899F0EC7671543B305FB5694024DE60A1B463D@medimmune4.medimmune.com> Does anyone out in Histoland have a recommendation for an off the shelf software program that we could use to accession case in the lab and that can be validated for GLP purposes? We will require it to accession specimens and place in a freezer for now, but would like it to double up as a regular Histology accessioning program. Also does anyone have an SOP on freezer maintenace they would be willing to share? This is also a GLP related issue. Thanks in advance. Nick Madary HT/HTL(ASCP)QIHC Histology Manager, Medimmune Inc Wk 301.398.6113 Madaryj@medimmune.com From immrstambo <@t> hotmail.com Fri Dec 10 19:57:31 2004 From: immrstambo <@t> hotmail.com (Christine Tambasco) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] recycled alcohol Message-ID: Does anyone have any experience with the CBG recycler and the Leica tissue processor? My recycled alcohol seems to have clear rite in it. The rep says it could be fat from the types of tissue processed and not clear rite at all. The recycled alcohol turns cloudly when mixed with water. I am relatively sure there isnt any clear rite in the alcohol before recycling, but I tested the alcohol before and after, and i seems to always be contaminated. I should think it couldnt be my processor (at least hope not) Thanks for any input. Christine Tambasco, HT (ASCP) St. Mary's Hospital Lab Amsterdam, New York From eileen_dusek <@t> yahoo.com Fri Dec 10 20:13:57 2004 From: eileen_dusek <@t> yahoo.com (eileen dusek) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] AFB staining Message-ID: <20041211021357.37667.qmail@web11907.mail.yahoo.com> Hi Everyone, We are having trouble with false positive AFB staining. Positive rods are staining on charged slides outside the tissue. The solution is filtered and run down on a Sakura H&E stainer. Any suggestions are very welcome. Eileen C. Dusek Edward Hospital --------------------------------- Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. From katri <@t> cogeco.ca Fri Dec 10 20:54:39 2004 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] AFB staining References: <20041211021357.37667.qmail@web11907.mail.yahoo.com> Message-ID: <000401c4df2c$c23b2e70$6a9a9618@Katri> Hi Eileen, Do your slides at any point get into tap water? Either in the waterbath or in the staining procedure? Tap water can have AFB positive bacteria, that could easily contaminate your slides. Just a thought.... Katri Katri Tuomala St.Joseph's Healthcare Hamilton, Ontario, Canada ----- Original Message ----- From: "eileen dusek" To: "histonet" Sent: Friday, December 10, 2004 9:13 PM Subject: [Histonet] AFB staining > Hi Everyone, > We are having trouble with false positive AFB staining. Positive rods are > staining on charged slides outside the tissue. The solution is filtered > and run down on a Sakura H&E stainer. > Any suggestions are very welcome. > > Eileen C. Dusek > Edward Hospital > > > --------------------------------- > Do you Yahoo!? > Yahoo! Mail - 250MB free storage. Do more. Manage less. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From bhewlett <@t> cogeco.ca Sat Dec 11 08:25:30 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Microtomy References: Message-ID: <001d01c4df8d$45ed7830$6400a8c0@mainbox> Luis, American Optical Scientific Instrument Division used to supply a great little book called 'The Effective Use and Proper Care of The Microtome' by Oscar W. Richards. It has all the gory details you're looking for!! I still use the diagrams in my copy for teaching. I haven't seen a new copy since the 70's, but you may be able to find one from a used book supplier. Bryan ----- Original Message ----- From: "Luis Chiriboga" To: "Histonet" Sent: Friday, December 10, 2004 5:12 PM Subject: [Histonet] Microtomy > Hi everyone > > Does anyone know of a text that describes (in detail) the physics and > geometry of tissue sectioning? I'm looking for all the gory details....... > > Thanks > > Luis > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Barry.R.Rittman <@t> uth.tmc.edu Sat Dec 11 11:50:32 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Microtomy Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0F352CC@UTHEVS3.mail.uthouston.edu> Luis The American Optical Booklet is very good. However I would also suggest the more comprehensive book by Steedman. Section Cutting in Microscopy. H.F. Steedman. Blackwell Scientific Publications. 1960. Barry From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan Hewlett Sent: Sat 12/11/2004 8:25 AM To: Luis Chiriboga; Histonet Subject: Re: [Histonet] Microtomy Luis, American Optical Scientific Instrument Division used to supply a great little book called 'The Effective Use and Proper Care of The Microtome' by Oscar W. Richards. It has all the gory details you're looking for!! I still use the diagrams in my copy for teaching. I haven't seen a new copy since the 70's, but you may be able to find one from a used book supplier. Bryan ----- Original Message ----- From: "Luis Chiriboga" To: "Histonet" Sent: Friday, December 10, 2004 5:12 PM Subject: [Histonet] Microtomy > Hi everyone > > Does anyone know of a text that describes (in detail) the physics and > geometry of tissue sectioning? I'm looking for all the gory details....... > > Thanks > > Luis > _______________________________________________ > Histonet mailing list > Histonet@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 contact <@t> excaliburpathology.com Sat Dec 11 12:51:08 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Validated Accessioning Program, Freezer Message-ID: <20041211185108.5836.qmail@web50310.mail.yahoo.com> I wrote my own program in Microsoft Access and it has even passed CLIA certification for human tissue. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From TJJ <@t> Stowers-Institute.org Sat Dec 11 14:13:46 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Re: recycled alcohol Message-ID: Christine, Regardless of brand of commercial recycler, once there is xylene in your alcohol, you can no longer recycle it as alcohol. You could, however, recycle it as xylene, although most of it would end up in the waste cut since there is only a slight amount of xylene contamination. It would be better to dispose of it. Waste segregation is critical for proper alcohol recycling. Any alcohol that has xylene carryover (flushes from the processor and the alcohols immediately post-xylene for deparaffinization) should be disposed of and not put through the recycler. You will never get the xylene out of the alcohol using an alcohol run. In smaller labs where there are only a few lab personnel, it's much easier to ensure proper waste segregation. In larger labs, with more hands in the pie, quite often someone dumps the bucket into the wrong container. Good luck! Teri Johnson Managing Director, Histology Facility Stowers Institute for Medical Research From clarke.ian <@t> virgin.net Sun Dec 12 15:37:09 2004 From: clarke.ian <@t> virgin.net (ian clarke) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Looking for a training procedure for BMS students Message-ID: Hi all, We have to set up new training procedures for BMS students in conjunction with the university so that they become ready for state registration . Has any one in the uk developed procedures that they would like to share? Are the students rotating through the different labs etc. hope you can help Ian clarke Cellular Pathology CAH Northern ireland From Kemlo.Rogerson <@t> elht.nhs.uk Mon Dec 13 03:05:28 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] recycled alcohol[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5D2@bhrv-nt-11.bhrv.nwest.nhs.uk> The cloudy alcohol 'test' tends to suggest it's contaminated with clearing agent. -----Original Message----- From: Christine Tambasco [mailto:immrstambo@hotmail.com] Sent: 11 December 2004 01:58 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] recycled alcohol[Scanned] Does anyone have any experience with the CBG recycler and the Leica tissue processor? My recycled alcohol seems to have clear rite in it. The rep says it could be fat from the types of tissue processed and not clear rite at all. The recycled alcohol turns cloudly when mixed with water. I am relatively sure there isnt any clear rite in the alcohol before recycling, but I tested the alcohol before and after, and i seems to always be contaminated. I should think it couldnt be my processor (at least hope not) Thanks for any input. Christine Tambasco, HT (ASCP) St. Mary's Hospital Lab Amsterdam, New York _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Dec 13 03:06:48 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] AFB staining[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5D3@bhrv-nt-11.bhrv.nwest.nhs.uk> There is an acid fast bacilli in water, but I can't remember for the life of me what it is. Anyone know? -----Original Message----- From: eileen dusek [mailto:eileen_dusek@yahoo.com] Sent: 11 December 2004 02:14 To: histonet Subject: [Histonet] AFB staining[Scanned] Hi Everyone, We are having trouble with false positive AFB staining. Positive rods are staining on charged slides outside the tissue. The solution is filtered and run down on a Sakura H&E stainer. Any suggestions are very welcome. Eileen C. Dusek Edward Hospital --------------------------------- Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eileen_dusek <@t> yahoo.com Mon Dec 13 07:06:46 2004 From: eileen_dusek <@t> yahoo.com (eileen dusek) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] AFB staining In-Reply-To: <000401c4df2c$c23b2e70$6a9a9618@Katri> Message-ID: <20041213130646.93299.qmail@web11904.mail.yahoo.com> There seems to be a common response, the tap water. We will run the slides down by hand. I appreciate all your help. Happy Holiday Eileen Katri Tuomala wrote: Hi Eileen, Do your slides at any point get into tap water? Either in the waterbath or in the staining procedure? Tap water can have AFB positive bacteria, that could easily contaminate your slides. Just a thought.... Katri Katri Tuomala St.Joseph's Healthcare Hamilton, Ontario, Canada ----- Original Message ----- From: "eileen dusek" To: "histonet" Sent: Friday, December 10, 2004 9:13 PM Subject: [Histonet] AFB staining > Hi Everyone, > We are having trouble with false positive AFB staining. Positive rods are > staining on charged slides outside the tissue. The solution is filtered > and run down on a Sakura H&E stainer. > Any suggestions are very welcome. > > Eileen C. Dusek > Edward Hospital > > > --------------------------------- > Do you Yahoo!? > Yahoo! Mail - 250MB free storage. Do more. Manage less. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Mail - Find what you need with new enhanced search. Learn more. From mprice26 <@t> juno.com Mon Dec 13 08:27:12 2004 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] RE: Glass Coverslippers and Automatic stainers Message-ID: <20041213.062720.19838.207644@webmail25.nyc.untd.com> Histonetters, I would like some feedback on what glass coverslippers you would recommend and autostainers. We are looking to purchase fairly soon and would like to set up an demo. Vendors are welcome to reply. Thank you. Marsha Price From gcallis <@t> montana.edu Mon Dec 13 09:37:16 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] AFB staining In-Reply-To: <20041211021357.37667.qmail@web11907.mail.yahoo.com> References: <20041211021357.37667.qmail@web11907.mail.yahoo.com> Message-ID: <6.0.0.22.1.20041213083044.01b37000@gemini.msu.montana.edu> You distilled water in waterbath may be the source. There are saprophitic Mycobacteria that grow in distilled water when you store your water in plastic carboys, and you pick up these bacteri when you pick up the sections. This has been published and also discussed in Histonet - check the archives. Be sure you wash plastic distilled water carboys frequently and use Clorox, followed by some good rinses. If distilled water is made in another area with huge storage tanks, the tanks should also be on a regular cleaning schedule including all tubing. I don't recall that the organisms come from tap water, but that also may be the case. At 07:13 PM 12/10/2004, you wrote: >Hi Everyone, >We are having trouble with false positive AFB staining. Positive rods are >staining on charged slides outside the tissue. The solution is filtered >and run down on a Sakura H&E stainer. >Any suggestions are very welcome. > >Eileen C. Dusek >Edward Hospital > > >--------------------------------- >Do you Yahoo!? > Yahoo! Mail - 250MB free storage. Do more. Manage less. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From SCheasty <@t> memorialcare.org Mon Dec 13 10:47:49 2004 From: SCheasty <@t> memorialcare.org (Sandy Cheasty) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Microwave Devices and Proposed Guidelines Message-ID: <59FE4C55358C6A42B84BAEC6092CEDBC05ACF547@sbnt7> If you receive any info on this would you please let me know. We are currently using a microwave from Wal-Mart. There has got to be some kind of exception somewhere. What about labs that could not justify a lab microwave but has a commercial one that works just as good for special stains only. Thanks, amy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sandy Cheasty Sent: Friday, December 10, 2004 2:58 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] Microwave Devices and Proposed Guidelines I know there are thousands of histology labs that use commercial microwaves safely and have used them safely for years. Does everyone: -violate OSHA? -lie to their lab managers? There must be some way of continuing to use commercial microwave ovens for special stains without being subversive. Don't get me wrong, I usually enjoy being subversive. Can someone please give me a justification for using a commercial microwave or tell me where I can get an OSHA approved one for less than $1000? Thanks again, Elvis -----Original Message----- From: Willis, Donna [mailto:DonnaWillis@texashealth.org] Sent: Friday, December 10, 2004 7:08 AM To: Sandy Cheasty; histonet@pathology.swmed.edu Subject: RE: [Histonet] NCCLS Microwave Device Use Proposed Guideline It is against OSHA guideline (OSHA 29 CFR 1910.303(b)(2))to use a commercial microwave (exp. Wal-Mart). The warranty of commercial units state that the instrument is not to be used for anything except its original use, food preparation. The OSHA guideline would include any equipment in the lab. If someone gets injured the lab would be at fault not the manufacture. Donna Willis Histology Lab Manager Harris Methodist Fort Worth,Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sandy Cheasty Sent: Friday, December 10, 2004 7:40 AM To: histonet@pathology.swmed.edu Subject: [Histonet] NCCLS Microwave Device Use Proposed Guideline I have the GP28-P guideline and read it but am unsure about its recommendations or limitations in using a microwave for special stains only. Is there any consensus among histology labs as to its interpretation? Is it still OK to use a commercial microwave from Wal-Mart without special exhaust system as long as you don't microwave formaldehyde, osmium tetroxide, lead acetate etc. (Paragraph 4.4) and do annual leakage tests? Thanks! Elvis ____________________________________________________________________________ __ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. ____________________________________________________________________________ __ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or an agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ______________________________________________________________________________ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm From JCarpenter764 <@t> aol.com Mon Dec 13 11:08:28 2004 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] fall practical???? Message-ID: <5430A13C.22A7F876.40C6E687@aol.com> Hey it's Jennell again.....I was just wondering if anyone has received their practical exam results... From cormier <@t> MIT.EDU Mon Dec 13 12:07:43 2004 From: cormier <@t> MIT.EDU (Kathy Cormier) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] galectin 4 Message-ID: <5.2.1.1.2.20041213130230.01557230@po14.mit.edu> Hello All, Hope your day is warm, our temp is rapidly dropping! I have a researcher who is looking for an antibody to work on FFPE mouse tissue, for galectine 4. Any one using anything that works for FFPE, not just Westerns/flow? :) Thanks! Kathy Cormier Div Comp Medicine MIT From MDiCarlo <@t> KaleidaHealth.Org Mon Dec 13 13:01:22 2004 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:24:24 2005 Subject: FW: [Histonet] Agitation of cassettes Message-ID: Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: DiCarlo, Margaret Sent: Friday, November 12, 2004 15:20 To: 'Patsy Ruegg' Subject: RE: [Histonet] Agitation of cassettes Patsy, I too also process most of my bone manually. I agitate on a magnetic stir plate at room temperature with the bone sitting on a piece of plastic or something of that nature. I never used it for fixation. I just usually let it sit in formalin for 5-7 days depending on size and thickness. How much less time are we talking about, with a bone section that is around 5-8mm in thickness? Thanks for your response. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Friday, November 12, 2004 12:36 To: 'Molinari, Betsy'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Agitation of cassettes I too have always processed most of my tissue by hand (usually bone) and swear by using a shaker for all steps, fixation, decal, etc. For mass processing using a tissue processor mine has a stir bar at the bottom of the tank and I do choose vacuum when ever possible. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Molinari, Betsy Sent: Friday, November 12, 2004 6:08 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Agitation of cassettes Hi Fran, Happy Friday to you! We do put our cassettes on a shaker. I work in a research lab and have the luxury of time (well, most of the time I do) to do so. I usually just keep the cassettes on overnight. I always put my decals on a rotator or shaker it cuts my decal time by quite a bit. I do not have a publication, but I am someone on this site will! Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fran Lemons Sent: Thursday, November 11, 2004 2:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Agitation of cassettes) Hello all, it's almost Friday!! Has anyone out there ever heard of placing the basket of cassettes in formalin on an agitator to stir the formalin as a way to expedite fixation? If so, can you direct me to something published on the subject? Also, has anyone heard of the same thing for bunions in decal solution? Thanks ahead of time, Fran Walker _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From RCHIOVETTI <@t> aol.com Mon Dec 13 13:38:12 2004 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Survey: Histo in Nevada? Message-ID: <8c.1c0ae154.2eef49a4@aol.com> Fellow Histonetters, If you are a practicing histotechnologist in the state of Nevada and if you can spare about 10-15 minutes to participate in a survey, I would appreciate hearing from you. This isn't a salary survey; it's geared more towards the general "health and welfare" of the profession and the labs in the state (size of labs, number of docs on staff, case load, instrumentation, usage, service and maintenance issues and so on). I need input from *all* histo labs, whether they're hospital labs, derm/Mohs clinics, veterinary diagnostics labs, reference labs, academic/research labs, etc. You can easily complete the survey at your computer, then simply return it to me as an attachment to an e-mail message. I also have a hard copy version that I can send to you via snail-mail if you prefer. Please contact me off-list if you'd like to participate or if you have any questions. Thanks in advance. Happy Holidays to everyone! Cheers, Robert (Bob) Chiovetti, Ph.D. Tucson, AZ From cfavara <@t> niaid.nih.gov Mon Dec 13 13:54:17 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] CD 68 on murine brain Message-ID: Has anyone done any CD68 oh FFPE mouse brain? I will search the archives and do a lit search but have not as of this time, Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From liz <@t> premierlab.com Mon Dec 13 14:39:04 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] CD 68 on murine brain In-Reply-To: Message-ID: <000201c4e153$c9489990$76d48a80@AMY> Cynthia I use serotec's antibody F4/80 for macrophages in mouse tissues. I have not tried it on brain, but I even have done it on formalin fixed, formic acid decalcifed mouse joints. This antibody will work on FFPE or paraformaldehye fixed tissue with proteinase K digestion. Here is the basic protocol. 1:1000 primary 30 minutes room temp Proteinase K pretreatment - 5 minutes Rabbit anti-rat 1:400 30 minutes Envision Rabbit - 30 minutes Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Favara, Cynthia (NIH/NIAID) Sent: Monday, December 13, 2004 12:54 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD 68 on murine brain Has anyone done any CD68 oh FFPE mouse brain? I will search the archives and do a lit search but have not as of this time, Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NBerghoff <@t> cvm.tamu.edu Mon Dec 13 14:52:30 2004 From: NBerghoff <@t> cvm.tamu.edu (Nora Berghoff) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Histonet: Movat's stain - advice needed! Message-ID: Hello, I have a question about Russell's modification of Movat's pentachrome stain. (Ref.: Russell, HK, "A Modification of Movat's Pentachrome stain", Arch Path, Vol. 94, Aug 1972, 187-191) I have been staining canine intestinal sections that have been fixed in 10% neutral buffered formalin, embedded in paraffin and cut at 5um. The stain worked pretty well and gave good results for a while. Last week I have had the following problem: After destaining the crocein scarlet-acid fuchsin with 5% phosphotungstic acid and then rinsing it in 0.5% acetic acid, it kept destaining the tissue until everything was just very pale red (one slide was completely destained - no red stain left whatsoever and the other stains were pale as well). I always control the destaining under the microscope and stop when there is still intense red color. The color kept coming off the slide in the acetic acid solution and also in the following rinses in absolute alcohol. I make the solutions (phosphotungstic acid and acetic acid) fresh right before I need them. Does anyone have an idea as to what the problem might be? Is the acetic acid not strong enough/the phosphotungstic acid too strong, etc?? Is it a pH problem? I have searched for an answer in histo books, on the internet and in the histonet archives, but couldn't find anything helpful... Any advice (even if it is just an idea) would be greatly appreciated!!! Thank you so much, Nora Berghoff Research Assistant Texas A&M University College Station, TX From JQB7 <@t> CDC.GOV Mon Dec 13 16:49:09 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Microwave Devices and Proposed Guidelines Message-ID: We have a commercial (for laboratory use) microwave in our main lab. Recently we set up a small, research lab for prion work. I needed a microwave for antigen retrieval but did not have space (or money) for a commercial microwave. Our safety department approved our use of a small, litchen microwave (we got it for less than $100.00 from Target) for this purpose. Jeanine Bartlett CDC, Atlanta -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Sandy Cheasty Sent: Fri 12/10/2004 2:58 PM To: 'histonet@pathology.swmed.edu' Cc: Subject: [Histonet] Microwave Devices and Proposed Guidelines I know there are thousands of histology labs that use commercial microwaves safely and have used them safely for years. Does everyone: -violate OSHA? -lie to their lab managers? There must be some way of continuing to use commercial microwave ovens for special stains without being subversive. Don't get me wrong, I usually enjoy being subversive. Can someone please give me a justification for using a commercial microwave or tell me where I can get an OSHA approved one for less than $1000? Thanks again, Elvis -----Original Message----- From: Willis, Donna [mailto:DonnaWillis@texashealth.org] Sent: Friday, December 10, 2004 7:08 AM To: Sandy Cheasty; histonet@pathology.swmed.edu Subject: RE: [Histonet] NCCLS Microwave Device Use Proposed Guideline It is against OSHA guideline (OSHA 29 CFR 1910.303(b)(2))to use a commercial microwave (exp. Wal-Mart). The warranty of commercial units state that the instrument is not to be used for anything except its original use, food preparation. The OSHA guideline would include any equipment in the lab. If someone gets injured the lab would be at fault not the manufacture. Donna Willis Histology Lab Manager Harris Methodist Fort Worth,Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sandy Cheasty Sent: Friday, December 10, 2004 7:40 AM To: histonet@pathology.swmed.edu Subject: [Histonet] NCCLS Microwave Device Use Proposed Guideline I have the GP28-P guideline and read it but am unsure about its recommendations or limitations in using a microwave for special stains only. Is there any consensus among histology labs as to its interpretation? Is it still OK to use a commercial microwave from Wal-Mart without special exhaust system as long as you don't microwave formaldehyde, osmium tetroxide, lead acetate etc. (Paragraph 4.4) and do annual leakage tests? Thanks! Elvis ____________________________________________________________________________ __ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. ______________________________________________________________________________ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From schnegelsberg <@t> xgene.com Mon Dec 13 15:23:52 2004 From: schnegelsberg <@t> xgene.com (Birthe Schnegelsberg) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] storage of cryosections Message-ID: HI. Do I actually have to add Drierite absorbent into the boxes with the cryo sections in the -20C freezer to avoid freezing artefacts? Thanks, birthe From joeamateur <@t> hotmail.com Mon Dec 13 18:50:10 2004 From: joeamateur <@t> hotmail.com (Jack England) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Collagen IHC in mouse tissue--need antibody recommendations! Message-ID: Aloha Histonetters, and happy holidays to all! We've been trying to do immunostaining for collagen I on some murine-derived cell lines, with next to no success. I'm almost certain that I've got collagen in there (PSR shows plenty of red, my poor-man's polarizers make the red stuff look birefringent, and SEM images show all kinds of fibrous ECM), but it's not staining for collagen I or III with the antibodies we've got (both from Sigma, produced in mouse ascites, not tested on mouse tissue). I suspect that our antibodies don't react with mouse tissue, but it's possible that the fixation (we use 10% NBF) or the embedding (paraffin, which does involve heating the tissue) could be denaturing the antigenic sites. I've tried enzymatic antigen retrieval, and it didn't work...next step is to get ahold of mouse tails and a cryostat and try for a positive control. My question for anyone that can help is, assuming that the problem is that our antibodies do not react with mouse tissue, can any of you recommend antibodies to collagen I or III that do react with mouse tissue (preferably FFPE mouse tissue, since we don't have a cryostat)? I've checked the U of Iowa hybridoma bank and come up empty, and a Biocompare search gave me one hit--I'm curious if any of you out there can recommend something beyond that one. Any takers? --With warm regards and aloha, Jack England Tissue Genesis, Inc. http://www.tissuegenesis.com _________________________________________________________________ On the road to retirement? Check out MSN Life Events for advice on how to get there! http://lifeevents.msn.com/category.aspx?cid=Retirement From jkiernan <@t> uwo.ca Mon Dec 13 19:43:26 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] storage of cryosections References: Message-ID: <41BE453E.BA6D31CA@uwo.ca> Freezing artefacts (ice crystal holes) form while the tissue is being frozen, not during storage of sections. Once the holes are there, because of too-slow freezing, nothing can get rid of them. Drierite (or a similar desiccant) ensures that the stored sections are in a dry atmosphere. You should let the box warm to room temperature before you open it; otherwise water from the air will condense on the cold sections. If they are of unfixed tissue this will cause some damage, which may or may not matter depending on what you intend to do with the slides. John Kiernan London, Canada. ------------------------------------------------ Birthe Schnegelsberg wrote: > > HI. > Do I actually have to add Drierite absorbent into the boxes with the cryo > sections in the -20C freezer to avoid freezing artefacts? > Thanks, birthe > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From immrstambo <@t> hotmail.com Mon Dec 13 19:52:18 2004 From: immrstambo <@t> hotmail.com (Christine Tambasco) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] IHC ~ Tissue Falling Off Slides In-Reply-To: <8E7AD740937B954F947F0DB4467EFEE056E93D@mail.srhs-pa.org> Message-ID: I use poly-l-lysine slides for immunos (i make them myself) . Also i use fisher's probe slides and the citra retreival i do in the microwave. I put the slides in a plastic slide mailer with citra - microwave on high for 45sec - 1min then I add more citra and microwave 3 minutes at 50% power. It works for me. Good Luck! Christine Tambasco, HT (ASCP) St. Mary's Hospital , Amsterdam, New York ph-5188417287 >From: "Jones, Laura" >To: "Histonet (E-mail)" >Subject: [Histonet] IHC ~ Tissue Falling Off Slides >Date: Thu, 9 Dec 2004 11:03:38 -0500 > >Hello fellow Histonetters. > >We have suddenly started to have problems with tissue falling off slides >during IHC processing. We suspect the problem may be our slides, although >we have been using the same ones from Mercedes Medical for a while and they >have worked beautifully! We have contacted them, and they are investigating >and very kindly replacing our stock, but in the meantime, our Pathologist >has asked me to ask all of you where else we can find 25 X 75 silinated >slides. We use the Zymed ST-5050 automated stainer, and the 1 X 3 slides >are sometimes a bit to big to fit on the carousel. > >Just for additional info, we seem to be experiencing problems mostly with >the more alkaline retreival solutions - AR-10 (Biogenex) and EDTA (Zymed) >and Trilogy (Cell Marque). We use the Black & Decker steamer, and have >adjusted our times in retrieval from 75 minutes (!) to 20 minutes, per >Pathologist direction. We have also tried 20 minutes heating, then adding >the slides for 20 minutes of boiling, then allowing them to stand for 20-30 >minutes. > >Thanks in advance for all of your knowledge! > >The Histochicks at Sharon Regional > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pmarcum <@t> polysciences.com Tue Dec 14 08:28:24 2004 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Question to assist a someone off this list Message-ID: <000e01c4e1e9$2b4a0f80$7f00a8c0@PMARCUM2K> Good Morning, I have had several request for slides coated with different silanes or charges for people doing cell culture, plastics and some special histology stains. They are looking for negatively charged slides or special coatings. Do any of you on HistoNet know anything about these slides like where I can tell them to go or what type of coatings are available? I am sorry to ask this however, I don't know where else to go and as histologist I am more familiar with the traditional coatings and subbing materials. Thanks for the help or direction! Pamela A Marcum Histology/Microscopy Product Development Manager Polysciences, Inc. 400 Valley Road Warrington, PA 18976 Telephone: 800-523-2575???? Ext. 167 215-343-6484???? Ext. 167 Fax:? 800-343-3291 ???????? 215-343-0214 From peoshel <@t> wisc.edu Tue Dec 14 09:01:13 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] storage of cryosections In-Reply-To: <41BE453E.BA6D31CA@uwo.ca> References: <41BE453E.BA6D31CA@uwo.ca> Message-ID: I'm afraid I have to disagree with a portion of John's email: ice crystal damage can happen in storage. Ice crystal growth and refreezing occurs at temperatures above about -60 to -40 deg C, depending on the specifics of the tissue, local environment, and all the rest of the annoying biological realities we deal with. If tissues are frozen rapidly enough to prevent crystal formation, or to form only tiny crystals, the tissues/cells will experience crystal growth if they are warmed above the recrystallization temperature. Much of the damage is actually caused by dehydration as the growing ice crystals pull free water out of cells and intercellular spaces, but as crystal growth continues, it can form holes. The preventative is to store samples in -80 deg C freezers and to warm them too quickly for recrystallizaton and crystal growth to occur. But, there is an even neater trick: store the samples under vacuum in a -80 freezer with *lots* of desiccant (Lots -- no, more ... pile it in the chamber. More!*). Something with a high water capacity: best is 3-4 Angstrom (4 is better than 3, 5 is no good) molecular sieve, next best is silica gel. Leave the samples like this for a few days (hours to weeks, depending on sample size and number -- it's empirical, try it and see), and they will be nicely freeze-dried by vacuum sublimation. They can then be stored at any temperature with desiccant with no worries about ice crystals, or much of anything. On exposure to air, the samples will immediately start rehydrating, so they *must* be at room temperature before removing them from the desiccant chamber, and processed right away. Caveat: I don't think this would adversely affect antigen epitopes, but this should be tested. Phil * If you think you have enough, you don't. >Freezing artefacts (ice crystal holes) form while >the tissue is being frozen, not during storage of >sections. Once the holes are there, because of >too-slow freezing, nothing can get rid of them. > >Drierite (or a similar desiccant) ensures that >the stored sections are in a dry atmosphere. >You should let the box warm to room temperature >before you open it; otherwise water from the air >will condense on the cold sections. If they are of >unfixed tissue this will cause some damage, which may >or may not matter depending on what you intend to >do with the slides. > >John Kiernan >London, Canada. >------------------------------------------------ >Birthe Schnegelsberg wrote: >> >> HI. >> Do I actually have to add Drierite absorbent into the boxes with the cryo >> sections in the -20C freezer to avoid freezing artefacts? >> Thanks, birthe >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From Janet.Bonner <@t> FLHOSP.ORG Tue Dec 14 09:44:22 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Microwave Devices and Proposed Guidelines Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4127@fh2k093.fhmis.net> I've found that the issue of commercial vs industrial microwaves boils down to a ventilation problem( possible explosion or respiratory problem on a "stain" becoming an aerosol) as well as QA as far as temperature and time regulation. The microwave should be vented (ours is in the back) to the outside or a fume hood. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sandy Cheasty Sent: Monday, December 13, 2004 11:48 AM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] Microwave Devices and Proposed Guidelines If you receive any info on this would you please let me know. We are currently using a microwave from Wal-Mart. There has got to be some kind of exception somewhere. What about labs that could not justify a lab microwave but has a commercial one that works just as good for special stains only. Thanks, amy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sandy Cheasty Sent: Friday, December 10, 2004 2:58 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] Microwave Devices and Proposed Guidelines I know there are thousands of histology labs that use commercial microwaves safely and have used them safely for years. Does everyone: -violate OSHA? -lie to their lab managers? There must be some way of continuing to use commercial microwave ovens for special stains without being subversive. Don't get me wrong, I usually enjoy being subversive. Can someone please give me a justification for using a commercial microwave or tell me where I can get an OSHA approved one for less than $1000? Thanks again, Elvis -----Original Message----- From: Willis, Donna [mailto:DonnaWillis@texashealth.org] Sent: Friday, December 10, 2004 7:08 AM To: Sandy Cheasty; histonet@pathology.swmed.edu Subject: RE: [Histonet] NCCLS Microwave Device Use Proposed Guideline It is against OSHA guideline (OSHA 29 CFR 1910.303(b)(2))to use a commercial microwave (exp. Wal-Mart). The warranty of commercial units state that the instrument is not to be used for anything except its original use, food preparation. The OSHA guideline would include any equipment in the lab. If someone gets injured the lab would be at fault not the manufacture. Donna Willis Histology Lab Manager Harris Methodist Fort Worth,Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sandy Cheasty Sent: Friday, December 10, 2004 7:40 AM To: histonet@pathology.swmed.edu Subject: [Histonet] NCCLS Microwave Device Use Proposed Guideline I have the GP28-P guideline and read it but am unsure about its recommendations or limitations in using a microwave for special stains only. Is there any consensus among histology labs as to its interpretation? Is it still OK to use a commercial microwave from Wal-Mart without special exhaust system as long as you don't microwave formaldehyde, osmium tetroxide, lead acetate etc. (Paragraph 4.4) and do annual leakage tests? Thanks! Elvis ____________________________________________________________________________ __ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. 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Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From RitaR <@t> lexhealth.org Tue Dec 14 09:56:25 2004 From: RitaR <@t> lexhealth.org (Rita Riddle) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] unscribe Message-ID: _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From gcallis <@t> montana.edu Tue Dec 14 10:04:37 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] storage of cryosections In-Reply-To: References: Message-ID: <6.0.0.22.1.20041214084115.01b0b540@gemini.msu.montana.edu> Birthe, Initially, how you snap freeze is the first defense against freezing artefacts. Frozen section stored with a dessicant is a good idea keep the sections dry during storage. The storage container must stay closed to equilibrate FS to room temperature. Water condensation is the enemy. Be sure to air dry the sections before going into storage also. A black plastic 25 slide capacity box works well. Also, opening a box directly from freezer and taking a few sections out, then returning unused sections to freezer must be avoided due FS freeze thaw. Put as many slides as you need per storage container for a staining session. Drierite is anhydrous calcium chloride and has a tendency to exfoliate fine CaCl2dust all over everything including your sections which bothered me. Try 10 to 18 mesh silica gel, (Fisher# S161-500). The little beads have blue indicator so you know when dessicant is depleted. Put Silica gel in a nylon processing bags (Thermo Electron or StatLab) or embedding bags aka tea bags from Fisher, fold over top, and staple on fold the keep bags closed. Temperature is critical for storage - if you have -80C available, use that instead of -20C, and NEVER store sections in a self defrosting freezer found in a refrigerator, freeze dry cycle is very damaging. Hopefully you can store FS at temperature lower than -20C to preserve antigenicity over a long period of time. It may work for short term storage and robust antigens. At 02:23 PM 12/13/2004, you wrote: >HI. >Do I actually have to add Drierite absorbent into the boxes with the cryo >sections in the -20C freezer to avoid freezing artefacts? >Thanks, birthe > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From bjackson <@t> unipathllc.com Tue Dec 14 10:08:09 2004 From: bjackson <@t> unipathllc.com (Brianna Jackson) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Payscale Message-ID: Hello, Can histotechs working in labs with separate IHC and Histo departments tell me if you have the same payscale? I've been told that IHC techs (even HTL certified) cannot be paid the same as production bench techs. I was just wondering if this is how it works in other labs. Thanks for your time, Brianna Jackson, BS, QIHC Denver, CO 303-512-2220 From GauchV <@t> mail.amc.edu Tue Dec 14 10:09:48 2004 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Position Message-ID: I have a tech with 10 years of experience who will be relocating to the Scottsdale,Arizona region who is seeking a Histotech position in that area. Does anyone know of any openings? Please send any correspondence to this e-mail address as she is not currently able to access the Histonet. Thank you for your help. Vicki Gauch Albany Medical Center Albany, NY From gcallis <@t> montana.edu Tue Dec 14 10:20:04 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Special coatings for slides In-Reply-To: <000e01c4e1e9$2b4a0f80$7f00a8c0@PMARCUM2K> References: <000e01c4e1e9$2b4a0f80$7f00a8c0@PMARCUM2K> Message-ID: <6.0.0.22.1.20041214091203.01b4e368@gemini.msu.montana.edu> Pam, I believe Erie Scientific is doing some of this from what my Erie rep tells me. Erie website has 800 number for tech services. They have some new things for Microarray work. Also try Grace Bio-Labs, they have a website, and you can call them for detailed discussion. Be prepared for some pricey items. At 07:28 AM 12/14/2004, you wrote: >Good Morning, > >I have had several request for slides coated with different silanes or >charges for people doing cell culture, plastics and some special histology >stains. They are looking for negatively charged slides or special coatings. >Do any of you on HistoNet know anything about these slides like where I can >tell them to go or what type of coatings are available? > >I am sorry to ask this however, I don't know where else to go and as >histologist I am more familiar with the traditional coatings and subbing >materials. > >Thanks for the help or direction! > >Pamela A Marcum >Histology/Microscopy >Product Development Manager >Polysciences, Inc. >400 Valley Road >Warrington, PA 18976 >Telephone: 800-523-2575 Ext. 167 > 215-343-6484 Ext. 167 > >Fax: 800-343-3291 > 215-343-0214 > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From CCollins <@t> propathlab.com Tue Dec 14 10:21:33 2004 From: CCollins <@t> propathlab.com (Cory Collins) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Varicella-zoster Message-ID: Howdy!! I work for a reference lab in Dallas and we are having trouble finding control tissue for Varicella-zoster. Does anyone know of a vendor that sells control slides for Varcilla-zoster? Or maybe know someone that has some extra paraffin embedded tissue that is positive for Varicella-zoster. Thanks, Cory Cory Collins, HT (ASCP) QIHC Immunohistochemistry Supervisor ProPath 8267 Elmbrook Drive Suite 100 Dallas, TX 75247 214-638-2000 ext 2027 214-237-1730 Fax To learn more about ProPath, please visit http://www.ProPathLab.com ______________________________________________________________________________ This e-mail may contain confidential or privileged information. If you think you have received this e-mail in error, please advise the sender by reply e-mail and then delete this e-mail immediately. From MElliott <@t> mrl.ubc.ca Tue Dec 14 11:19:47 2004 From: MElliott <@t> mrl.ubc.ca (Mark Elliott) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] MMP-1 in Bouin's fixed tissue Message-ID: Has anyone had any experience staining for MMP-1 in Bouin's fixed human tissues? One of the Hospital's we collaborate with fixes their lungs in Bouin's. We have tried staining with no antigen retrieval and with/without overnight incubation in primary. Have also tried antigen retrieval using citra pH 6 in 95 C water bath and in autoclave with no luck. We are using MAB 1346 from Chemicon followed by APAAP technique. Are going to try high pH antigen retrieval and possibly trypsin but would appreciate any other suggestions. Thanks Mark Elliott iCAPTURE Centre Vancouver BC From contact <@t> excaliburpathology.com Tue Dec 14 11:34:31 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Collagen IHC in mouse tissue - need antibody recommendations Message-ID: <20041214173431.64936.qmail@web50308.mail.yahoo.com> Have you tried just a histochemical stain for collagen such as Masson's Trichrome? Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From Terry.Marshall <@t> rothgen.nhs.uk Tue Dec 14 11:38:18 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Collagen IHC in mouse tissue - need antibodyrecommendations Message-ID: Masson - histochemical? 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: Paula Pierce [mailto:contact@excaliburpathology.com] Sent: 14 December 2004 17:35 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Collagen IHC in mouse tissue - need antibodyrecommendations Have you tried just a histochemical stain for collagen such as Masson's Trichrome? Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dpahisto <@t> yahoo.com Tue Dec 14 12:09:38 2004 From: dpahisto <@t> yahoo.com (Cindy DuBois) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Ventana Benchmark Message-ID: <20041214180938.5019.qmail@web41305.mail.yahoo.com> Our lab is considering purchasing a Ventana Benchmark to do both immuno's and HPV testing (on thin preps). Has anyone had difficulty with this machine? With our volume we would have to do 2 runs a day, one for immunos - and one for HPV. I was also told we would begin doing ER/PR also. Where can I get the most valuable information regaring ER/PR staining? Are there any classes available? Cindy DuBois, HT ASCP DELTA PATHOLOGY ASSOC. STOCKTON CA --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com/a From gcallis <@t> montana.edu Tue Dec 14 12:09:42 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Which Collagen type for IHC in mouse tissue In-Reply-To: <20041214173431.64936.qmail@web50308.mail.yahoo.com> References: <20041214173431.64936.qmail@web50308.mail.yahoo.com> Message-ID: <6.0.0.22.1.20041214110534.01b265e8@gemini.msu.montana.edu> Which collagen do you need, Chemicon has Types 1 through IX, but Type IV polyclonal is the only one that they say cross reacts to mouse collagen. At 10:34 AM 12/14/2004, you wrote: >Have you tried just a histochemical stain for collagen such as Masson's >Trichrome? > > >Paula Pierce, HTL(ASCP)HT > >Excalibur Pathology, Inc. >631 N. Broadway >Moore, OK 73160 >405-759-3953 >contact@excaliburpathology.com >www.excaliburpathology.com Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Tue Dec 14 12:12:06 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Whoops, other collagen types for mouse IHC Message-ID: <6.0.0.22.1.20041214111110.01afc3b0@gemini.msu.montana.edu> Chemicon also has Type I and II reactive to mouse Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From jkiernan <@t> uwo.ca Tue Dec 14 12:42:20 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Which Collagen type for IHC in mouse tissue References: <20041214173431.64936.qmail@web50308.mail.yahoo.com> <6.0.0.22.1.20041214110534.01b265e8@gemini.msu.montana.edu> Message-ID: <41BF340C.6F4C4E50@uwo.ca> A recent (2004) publication by M.R.D'Andrea describes another way to unmask collagen for immunohistochemistry. Summary follows. The paper also includes some interesting information about collagenase. ______________________________ Publisher: Taylor & Francis Health Sciences, part of the Taylor & Francis Group http://journalsonline.tandf.co.uk Journal: Biotechnic & Histochemistry Issue: Volume 79, Number 2 / April 2004 Pages: 55 - 64 URL: Linking Options DOI: 10.1080/10520290410001728972 Collagenase predigestion on paraffin sections enhances collagen immunohistochemical detection without distorting tissue morphology Biotechnic & Histochemistry 79 (2) 55-64 by MR D'Andrea Johnson & Johnson Pharmaceutical Research and Development Drug Discovery Pennsylvania 19477-0776 Spring House Abstract: Reliable immunohistochemical detection of collagen in formalin fixed, paraffin embedded tissues requires protease digestion. While these pan-proteases (pepsin, trypsin, protease K, etc.) enhance collagen detection, they also digest many other tissue proteins and produce poor cellular morphology and unrecognizable cellular structures. Balancing the conditions (protease type, concentration, incubation time and temperature) to digest some, but not all, proteins in a tissue section while optimizing collagen detection requires one to compromise improved collagen immunolabeling with adequate cellular morphology. Furthermore, optimal conditions for digesting tissue proteins to enhance collagen detection vary among tissue types and their fixation. Although brain is not typically subject to these deleterious consequences, structures such as epithelium, spermatids, stroma etc. and other tissues with complicated histology are profoundly affected. To resolve this technical dilemma, we discovered a novel use for collagenase to enhance collagen immunodetection without affecting the noncollagen proteins, thereby preserving tissue morphology. Collagenase, which is typically used in vitro for disassociation of cells, has never been used reliably on formalin fixed, paraffin embedded tissue sections. This new use of collagenase for immunohistochemistry promotes increased collagen immunolabeling, is easy to use, is versatile, and allows preservation of tissue structure that provides maximal and accurate histological information. Keywords: Collagen, Collagenase, Over-digestion, Immunohistochemistry References: Alberts B, Bray D, Lewis J, Raff M, Roberts K, Watson JD The Molecular Biology of the Cell 1989; 808-8152nd ed Barsky SH, Roa NC, Restrepo C, Liotta LA Immunohistochemical enhancement of basement membrane antigens by pepsin: applications in diagnostic pathology Am. J. Clin. Pathol. 1984; 82: 191-194 Barsky SH, Siegal G, Jannotta F, Liotta LA Loss of basement membrane components by invasive tumors, but not by their benign counterparts Lab Invest. 1983; 49: 140-148 D'Andrea MR Evidence linking autoimmunity to neuronal cell death in Alzheimer's disease Brain Res. 2003; 982: 19-30 D'Andrea MR, Reiser PA, Gumula NA, Hertzog BM, Andrade-Gordon P Application of triple-label immunohistochemistry to characterize inflammation in Alzheimer's disease brains Biotech. & Histochem. 2001; 76: 97-106 D'Andrea MR, Reiser PA, Polkovitch DA, Gumula NA, Belkowski S, Branchide B, Hertzog BM, Schmidheiser D, Belkowski S, Gastard M, Andrade-Gordon P The use of formic acid to embellish amyloid plaque detection in Alzheimer's disease tissues misguides key observations Neurosci. Lett. 2003; 342: 114-118 Farquharson C, Robins SP Immunolocalization of collagen types I and III in the arterial wall of the rat Histochem. J. 1989; 21: 172-178 Grimaud JA, Druguet M, Emonard H, Peyrol S, Barioz CM, Guerret S Collagen immunotyping NATO ASI Series, Series A: Life Sciences 1985; 93: 487-514 Higuchi I, Suehara M, Iwaki H, Nakagawa M, Arimura K, Osame M Collagen VI deficiency Ullrich's disease Ann. Neurol. 2001; 49: 54 Kallioinen M, Autio-Harmainen H, Dammert K, Risteli J, Risteli L Basement membrane laminin and type IV collagen in various benign and malignant adnexal tumors of the skin: An immunohistochemical study J. Invest. Derm. 1984; 83: 276-280 Kasper M, Schuh D, Muller M Immunochemical localization of the ? subunit of prolyl 4-hydroxylase in human alveolar epithelial cells Acta Histochem. 1994; 96: 309-313 Liotta LA Tumor invasion and metastasis-role of the extracellular matrix: Rhoads Memorial Award Lecture Cancer Res. 1986; 46: 1-7 Lowry A, Wilcox D, Masson EA, Williams PE Immunohistochemical methods for semiquantitative analysis of collagen content in human peripheral nerve J. Anat. 1997; 191: 367-374 Rucklidge GJ, Dean V Immunochemical techniques in the study of protein degradation with particular reference to collagen Meth. Surv. Biochem. Anal. 1987; 17: 387-392 Sano J, Ishida A, Ishizaki M, Sugisaki Y, Yamanaka N, Masugi Y Immunofluorescent visualization of type IV collagen in the glomerular capillary basement membrane after digestion with a microbial protease Biomed. Res. 1984; 5: 387-392 Shi S-R, Cote RJ, Taylor CR Antigen retrieval immunohistochemistry: past, present, and future J. Histochem. Cytochem. 1997; 54: 327-343 Taylor CR, Shi S-R, Chaiwun B, Young L, Imam SA, Cote RJ Strategies for improving the immunohistochemical staining of various intranuclear prognostic markers in formalin-paraffin sections: androgen receptor, estrogen receptor, progesterone receptor, p53 protein, proliferating cell nuclear antigen, and Ki-67 antigen revealed by antigen retrieval technique Human Pathol. 1994; 25: 263-270 _______________ If your library subscribes to the Taylor & Francis journals package you can get the full text, complete with coloured pictures, on the web as a .pdf file. Go to: http://journalsonline.tandf.co.uk/app/home/contribution.asp?wasp=n2v6e330wl2wynff5d9l&referrer=parent&backto=issue,1,7;journal,2,17;searchpublicationsresults,1,1;homemain,1,1; (or you can get to it from http://journalsonline.tandf.co.uk) and then click on Open - Entire document. -- ------------------------------- 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 _______________________________ Gayle Callis wrote: > > Which collagen do you need, Chemicon has Types 1 through IX, but Type IV > polyclonal is the only one that they say cross reacts to mouse collagen. > > At 10:34 AM 12/14/2004, you wrote: > >Have you tried just a histochemical stain for collagen such as Masson's > >Trichrome? > > > > > >Paula Pierce, HTL(ASCP)HT > > > >Excalibur Pathology, Inc. > >631 N. Broadway > >Moore, OK 73160 > >405-759-3953 > >contact@excaliburpathology.com > >www.excaliburpathology.com > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From arunams <@t> interchange.ubc.ca Tue Dec 14 13:35:49 2004 From: arunams <@t> interchange.ubc.ca (arunams) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] MOMA antiboday Message-ID: <25912335.1103052949333.JavaMail.myubc2@portal9.itservices.ubc.ca> Hello, Is there anyone using MOMA antibody with good results. If so can you please share the information on where you get the Ab as well as what tissue preperation works best (Frozen or parafin) Thanks for your help. aruna From chris <@t> biocare.net Tue Dec 14 13:43:58 2004 From: chris <@t> biocare.net (Chris Von Vedder) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Job Posting: Biocare Medical Message-ID: <012401c4e215$414b48e0$1501a8c0@biocarechris> Biocare Medical has the following position open: Immunohistochemistry/Histologist: Requirements: Candidate must be a progressive histologist (HT or HTL) with a minimum of 5 to 10 years experience in immunohistochemistry techniques. The job requires a self-motivated individual with a passion for patient care. Both clinical and research experience is a plus, with emphasis on clinical immunohistochemistry. Job Duties: Product development, immunohistochemistry, titrations, double stains, in situ hybridization techniques, tissue microarrays and cutting formalin-fixed paraffin-embedded tissues. For additional information about BIOCARE MEDICAL, contact our website at www.biocare.net. If interested, please send your CV to dave@biocare.net or mail to Dr. David Tacha, VP of Immunohistochemistry, BIOCARE MEDICAL, 2940 Camino Diablo, Suite 300, Walnut Creek, CA 94597 From laurie.colbert <@t> huntingtonhospital.com Tue Dec 14 14:02:43 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] recycled alcohol Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5C167@EXCHANGE1.huntingtonhospital.com> Do you flush the tank with alcohol when you switch from Clear Rite to alcohol? Laurie Colbert -----Original Message----- From: Christine Tambasco [mailto:immrstambo@hotmail.com] Sent: Friday, December 10, 2004 5:58 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] recycled alcohol Does anyone have any experience with the CBG recycler and the Leica tissue processor? My recycled alcohol seems to have clear rite in it. The rep says it could be fat from the types of tissue processed and not clear rite at all. The recycled alcohol turns cloudly when mixed with water. I am relatively sure there isnt any clear rite in the alcohol before recycling, but I tested the alcohol before and after, and i seems to always be contaminated. I should think it couldnt be my processor (at least hope not) Thanks for any input. Christine Tambasco, HT (ASCP) St. Mary's Hospital Lab Amsterdam, New York _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joeamateur <@t> hotmail.com Tue Dec 14 15:08:10 2004 From: joeamateur <@t> hotmail.com (Jack England) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Which Collagen type for IHC in mouse tissue In-Reply-To: <6.0.0.22.1.20041214110534.01b265e8@gemini.msu.montana.edu> Message-ID: Wow. That was some response. Thanks, all! Per some of the questions asked, I have already done staining on my sections: Russell-Movat pentachrome shows my "collagen" structures to be yellow (which it should be), Verhoeff-van Gieson shows them to be red, and picro-sirius red shows them to be red as well. When viewed with crossed linear polarizers (unsolicited shout-out to Edmund Industrial Optics for their polarizer experimenter's kit!!), these structures are refractive, though I don't consider my knowledge thorough enough to call this "birefringence" or not. SEM images show my structures to be fibrous, and 60-70nm thick. All of this taken together indicates to me that I am looking at collagen (especially as my cells are ECM-secreting fibroblasts). Where it has gotten tricky is in trying to figure out what flavor of collagen I'm looking at. I am specifically interested in probing for collagens I, III, and IV, though it wouldn't be a bad idea to look at procollagen as well. At the moment, though, cash is an issue, so I'm looking chiefly for collagen I and III antibodies that react with murine tissue. Preferably antibodies that don't cross-react with other subtypes, but I'm not picky at this point. Thanks for all the suggestions...not sure what I'm going to do for a collagen III antibody, but I'm going to try a rabbit polyclonal anti-collagen I to see if that will work. In the meantime, I'm going to try to see if 1) I can sucker someone in the area out of some time with a cryostat, and 2) I can get ahold of a mouse tail for a positive control. Thanks again for all your help, and best wishes! --With aloha, Jack England Tissue Genesis, Inc. http://www.tissuegenesis.com _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From cwscouten <@t> myneurolab.com Tue Dec 14 15:30:24 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] storage of cryosections Message-ID: I agree that initial freezing is most important, but that freezing artifact can develop in storage because the ice will reform and crystalize. See the following link for a thorough discussion. http://www.myneurolab.com/global/Manuals/Tips%20and%20Techniques%20Freezing%20Artifact.pdf Drying agents are thus a good idea. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Philip Oshel Sent: Tuesday, December 14, 2004 9:01 AM To: jkiernan@uwo.ca Cc: Histonet@Pathology.swmed.edu Subject: Re: [Histonet] storage of cryosections I'm afraid I have to disagree with a portion of John's email: ice crystal damage can happen in storage. Ice crystal growth and refreezing occurs at temperatures above about -60 to -40 deg C, depending on the specifics of the tissue, local environment, and all the rest of the annoying biological realities we deal with. If tissues are frozen rapidly enough to prevent crystal formation, or to form only tiny crystals, the tissues/cells will experience crystal growth if they are warmed above the recrystallization temperature. Much of the damage is actually caused by dehydration as the growing ice crystals pull free water out of cells and intercellular spaces, but as crystal growth continues, it can form holes. The preventative is to store samples in -80 deg C freezers and to warm them too quickly for recrystallizaton and crystal growth to occur. But, there is an even neater trick: store the samples under vacuum in a -80 freezer with *lots* of desiccant (Lots -- no, more ... pile it in the chamber. More!*). Something with a high water capacity: best is 3-4 Angstrom (4 is better than 3, 5 is no good) molecular sieve, next best is silica gel. Leave the samples like this for a few days (hours to weeks, depending on sample size and number -- it's empirical, try it and see), and they will be nicely freeze-dried by vacuum sublimation. They can then be stored at any temperature with desiccant with no worries about ice crystals, or much of anything. On exposure to air, the samples will immediately start rehydrating, so they *must* be at room temperature before removing them from the desiccant chamber, and processed right away. Caveat: I don't think this would adversely affect antigen epitopes, but this should be tested. Phil * If you think you have enough, you don't. >Freezing artefacts (ice crystal holes) form while the tissue is being >frozen, not during storage of sections. Once the holes are there, >because of too-slow freezing, nothing can get rid of them. > >Drierite (or a similar desiccant) ensures that the stored sections are >in a dry atmosphere. >You should let the box warm to room temperature before you open it; >otherwise water from the air will condense on the cold sections. If >they are of unfixed tissue this will cause some damage, which may or >may not matter depending on what you intend to do with the slides. > >John Kiernan >London, Canada. >------------------------------------------------ >Birthe Schnegelsberg wrote: >> >> HI. >> Do I actually have to add Drierite absorbent into the boxes with the >> cryo sections in the -20C freezer to avoid freezing artefacts? >> Thanks, birthe >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joeamateur <@t> hotmail.com Tue Dec 14 21:07:22 2004 From: joeamateur <@t> hotmail.com (Jack England) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Which Collagen type for IHC in mouse tissue In-Reply-To: <003c01c4e23b$3e501500$eefc6c44@sd.cox.net> Message-ID: Unfortunately no, I'm not. (I may get flamed for this next part, but:) I wish I were seasoned enough at this histo alchemy to know what I was doing (let alone interpret what I get!), but unfortunately I'm just a young, brand-new rookie at all of it, formerly an antibody manufacturer, now a research tissue engineer. But please do say more. Now the world and I are curious. *grin* That said, I worked with a Sarah Jones from CA during a summer in Zimbabwe...wouldn't be you, would it? --Aloha, Jack England Tissue Genesis, Inc. http://www.tissuegenesis.com >From: "Sarah Jones" >To: "Jack England" >Subject: Re: [Histonet] Which Collagen type for IHC in mouse tissue >Date: Tue, 14 Dec 2004 16:15:54 -0800 > >Okay, okay....I have been wanting to ask this for some time. Are you the >Pathologist, Jack England, who was once-upon-a-time with MLS in Orange Co., >CA., and then more recently with CHOMP in Monterey, CA.? I need to know >the >answer to this before I say any more. > >Thanks! >Sarah "Sally" Jones, HT/HTL (ASCP) >now working for DakoCytomation in Carpinteria, CA. _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From arrrid702 <@t> skmc.gov.ae Tue Dec 14 22:26:45 2004 From: arrrid702 <@t> skmc.gov.ae (Ridhwaan Arries) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] ihc problem on these Dako HSV-1,HSV-2,HBSAG Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E02D19222@SKMCEMAIL.skmc.gov.ae> Good morning, Could any ihc experts out there please help me in solving these problems 1. Dako HSV-1 (clone B0114) The dilution I used was 1:100-Too much background staining on Dako HSV CONTROL SLIDES (code #T1150).Slides were HIER and tried without HIER With the same results. 2. Dako HSV-2 (clone B0116) The dilution I used was 1:100-Too much background staining on Dako HSV CONTROL SLIDES (code #T1150).Slides were treated as above. 3. Dako HBsAG (clone B0560) The dilution used was 1:15,000-No staining at all on Dako HBsAG CONTROL Slides (code #T1110).Slides were pretreated with and without HIER as well All enzymes. Anyone up to advice? Ridhwaan Arries-Med.Tech./HISTOLOGY(Pentech-South Africa) -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Tuesday, December 14, 2004 10:14 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 13, Issue 20 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. galectin 4 (Kathy Cormier) 2. FW: [Histonet] Agitation of cassettes (DiCarlo, Margaret) 3. Survey: Histo in Nevada? (RCHIOVETTI@aol.com) 4. CD 68 on murine brain (Favara, Cynthia (NIH/NIAID)) 5. RE: CD 68 on murine brain (Elizabeth Chlipala) 6. Histonet: Movat's stain - advice needed! (Nora Berghoff) 7. RE: Microwave Devices and Proposed Guidelines (Bartlett, Jeanine) 8. storage of cryosections (Birthe Schnegelsberg) 9. Collagen IHC in mouse tissue--need antibody recommendations! (Jack England) 10. Re: storage of cryosections (John Kiernan) 11. RE: IHC ~ Tissue Falling Off Slides (Christine Tambasco) 12. Question to assist a someone off this list (Pamela Marcum) 13. Re: storage of cryosections (Philip Oshel) 14. RE: Microwave Devices and Proposed Guidelines (Bonner, Janet) 15. unscribe (Rita Riddle) 16. Re: storage of cryosections (Gayle Callis) 17. Payscale (Brianna Jackson) 18. Position (Vicki Gauch) 19. Special coatings for slides (Gayle Callis) 20. Varicella-zoster (Cory Collins) 21. MMP-1 in Bouin's fixed tissue (Mark Elliott) 22. Collagen IHC in mouse tissue - need antibody recommendations (Paula Pierce) 23. RE: Collagen IHC in mouse tissue - need antibodyrecommendations (Marshall Terry Dr, Consultant Histopathologist) ---------------------------------------------------------------------- Message: 1 Date: Mon, 13 Dec 2004 13:07:43 -0500 From: Kathy Cormier Subject: [Histonet] galectin 4 To: histonet@lists.utsouthwestern.edu Message-ID: <5.2.1.1.2.20041213130230.01557230@po14.mit.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Hello All, Hope your day is warm, our temp is rapidly dropping! I have a researcher who is looking for an antibody to work on FFPE mouse tissue, for galectine 4. Any one using anything that works for FFPE, not just Westerns/flow? :) Thanks! Kathy Cormier Div Comp Medicine MIT ------------------------------ Message: 2 Date: Mon, 13 Dec 2004 14:01:22 -0500 From: "DiCarlo, Margaret" Subject: FW: [Histonet] Agitation of cassettes To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: DiCarlo, Margaret Sent: Friday, November 12, 2004 15:20 To: 'Patsy Ruegg' Subject: RE: [Histonet] Agitation of cassettes Patsy, I too also process most of my bone manually. I agitate on a magnetic stir plate at room temperature with the bone sitting on a piece of plastic or something of that nature. I never used it for fixation. I just usually let it sit in formalin for 5-7 days depending on size and thickness. How much less time are we talking about, with a bone section that is around 5-8mm in thickness? Thanks for your response. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Friday, November 12, 2004 12:36 To: 'Molinari, Betsy'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Agitation of cassettes I too have always processed most of my tissue by hand (usually bone) and swear by using a shaker for all steps, fixation, decal, etc. For mass processing using a tissue processor mine has a stir bar at the bottom of the tank and I do choose vacuum when ever possible. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Molinari, Betsy Sent: Friday, November 12, 2004 6:08 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Agitation of cassettes Hi Fran, Happy Friday to you! We do put our cassettes on a shaker. I work in a research lab and have the luxury of time (well, most of the time I do) to do so. I usually just keep the cassettes on overnight. I always put my decals on a rotator or shaker it cuts my decal time by quite a bit. I do not have a publication, but I am someone on this site will! Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fran Lemons Sent: Thursday, November 11, 2004 2:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Agitation of cassettes) Hello all, it's almost Friday!! Has anyone out there ever heard of placing the basket of cassettes in formalin on an agitator to stir the formalin as a way to expedite fixation? If so, can you direct me to something published on the subject? Also, has anyone heard of the same thing for bunions in decal solution? Thanks ahead of time, Fran Walker _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. ------------------------------ Message: 3 Date: Mon, 13 Dec 2004 14:38:12 EST From: RCHIOVETTI@aol.com Subject: [Histonet] Survey: Histo in Nevada? To: histonet@lists.utsouthwestern.edu Message-ID: <8c.1c0ae154.2eef49a4@aol.com> Content-Type: text/plain; charset="US-ASCII" Fellow Histonetters, If you are a practicing histotechnologist in the state of Nevada and if you can spare about 10-15 minutes to participate in a survey, I would appreciate hearing from you. This isn't a salary survey; it's geared more towards the general "health and welfare" of the profession and the labs in the state (size of labs, number of docs on staff, case load, instrumentation, usage, service and maintenance issues and so on). I need input from *all* histo labs, whether they're hospital labs, derm/Mohs clinics, veterinary diagnostics labs, reference labs, academic/research labs, etc. You can easily complete the survey at your computer, then simply return it to me as an attachment to an e-mail message. I also have a hard copy version that I can send to you via snail-mail if you prefer. Please contact me off-list if you'd like to participate or if you have any questions. Thanks in advance. Happy Holidays to everyone! Cheers, Robert (Bob) Chiovetti, Ph.D. Tucson, AZ ------------------------------ Message: 4 Date: Mon, 13 Dec 2004 14:54:17 -0500 From: "Favara, Cynthia (NIH/NIAID)" Subject: [Histonet] CD 68 on murine brain To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Has anyone done any CD68 oh FFPE mouse brain? I will search the archives and do a lit search but have not as of this time, Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives ------------------------------ Message: 5 Date: Mon, 13 Dec 2004 13:39:04 -0700 From: "Elizabeth Chlipala" Subject: RE: [Histonet] CD 68 on murine brain To: "'Favara, Cynthia \(NIH/NIAID\)'" , Message-ID: <000201c4e153$c9489990$76d48a80@AMY> Content-Type: text/plain; charset="US-ASCII" Cynthia I use serotec's antibody F4/80 for macrophages in mouse tissues. I have not tried it on brain, but I even have done it on formalin fixed, formic acid decalcifed mouse joints. This antibody will work on FFPE or paraformaldehye fixed tissue with proteinase K digestion. Here is the basic protocol. 1:1000 primary 30 minutes room temp Proteinase K pretreatment - 5 minutes Rabbit anti-rat 1:400 30 minutes Envision Rabbit - 30 minutes Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Favara, Cynthia (NIH/NIAID) Sent: Monday, December 13, 2004 12:54 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD 68 on murine brain Has anyone done any CD68 oh FFPE mouse brain? I will search the archives and do a lit search but have not as of this time, Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Mon, 13 Dec 2004 14:52:30 -0600 From: "Nora Berghoff" Subject: [Histonet] Histonet: Movat's stain - advice needed! To: Message-ID: Content-Type: text/plain; charset=US-ASCII Hello, I have a question about Russell's modification of Movat's pentachrome stain. (Ref.: Russell, HK, "A Modification of Movat's Pentachrome stain", Arch Path, Vol. 94, Aug 1972, 187-191) I have been staining canine intestinal sections that have been fixed in 10% neutral buffered formalin, embedded in paraffin and cut at 5um. The stain worked pretty well and gave good results for a while. Last week I have had the following problem: After destaining the crocein scarlet-acid fuchsin with 5% phosphotungstic acid and then rinsing it in 0.5% acetic acid, it kept destaining the tissue until everything was just very pale red (one slide was completely destained - no red stain left whatsoever and the other stains were pale as well). I always control the destaining under the microscope and stop when there is still intense red color. The color kept coming off the slide in the acetic acid solution and also in the following rinses in absolute alcohol. I make the solutions (phosphotungstic acid and acetic acid) fresh right before I need them. Does anyone have an idea as to what the problem might be? Is the acetic acid not strong enough/the phosphotungstic acid too strong, etc?? Is it a pH problem? I have searched for an answer in histo books, on the internet and in the histonet archives, but couldn't find anything helpful... Any advice (even if it is just an idea) would be greatly appreciated!!! Thank you so much, Nora Berghoff Research Assistant Texas A&M University College Station, TX ------------------------------ Message: 7 Date: Mon, 13 Dec 2004 17:49:09 -0500 From: "Bartlett, Jeanine" Subject: RE: [Histonet] Microwave Devices and Proposed Guidelines To: "Sandy Cheasty" , Message-ID: Content-Type: text/plain; charset="UTF-8" We have a commercial (for laboratory use) microwave in our main lab. Recently we set up a small, research lab for prion work. I needed a microwave for antigen retrieval but did not have space (or money) for a commercial microwave. Our safety department approved our use of a small, litchen microwave (we got it for less than $100.00 from Target) for this purpose. Jeanine Bartlett CDC, Atlanta -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Sandy Cheasty Sent: Fri 12/10/2004 2:58 PM To: 'histonet@pathology.swmed.edu' Cc: Subject: [Histonet] Microwave Devices and Proposed Guidelines I know there are thousands of histology labs that use commercial microwaves safely and have used them safely for years. Does everyone: -violate OSHA? -lie to their lab managers? There must be some way of continuing to use commercial microwave ovens for special stains without being subversive. Don't get me wrong, I usually enjoy being subversive. Can someone please give me a justification for using a commercial microwave or tell me where I can get an OSHA approved one for less than $1000? Thanks again, Elvis -----Original Message----- From: Willis, Donna [mailto:DonnaWillis@texashealth.org] Sent: Friday, December 10, 2004 7:08 AM To: Sandy Cheasty; histonet@pathology.swmed.edu Subject: RE: [Histonet] NCCLS Microwave Device Use Proposed Guideline It is against OSHA guideline (OSHA 29 CFR 1910.303(b)(2))to use a commercial microwave (exp. Wal-Mart). The warranty of commercial units state that the instrument is not to be used for anything except its original use, food preparation. The OSHA guideline would include any equipment in the lab. If someone gets injured the lab would be at fault not the manufacture. Donna Willis Histology Lab Manager Harris Methodist Fort Worth,Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sandy Cheasty Sent: Friday, December 10, 2004 7:40 AM To: histonet@pathology.swmed.edu Subject: [Histonet] NCCLS Microwave Device Use Proposed Guideline I have the GP28-P guideline and read it but am unsure about its recommendations or limitations in using a microwave for special stains only. Is there any consensus among histology labs as to its interpretation? Is it still OK to use a commercial microwave from Wal-Mart without special exhaust system as long as you don't microwave formaldehyde, osmium tetroxide, lead acetate etc. (Paragraph 4.4) and do annual leakage tests? Thanks! Elvis ____________________________________________________________________________ __ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. ______________________________________________________________________________ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Mon, 13 Dec 2004 15:23:52 -0600 From: "Birthe Schnegelsberg" Subject: [Histonet] storage of cryosections To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" HI. Do I actually have to add Drierite absorbent into the boxes with the cryo sections in the -20C freezer to avoid freezing artefacts? Thanks, birthe ------------------------------ Message: 9 Date: Mon, 13 Dec 2004 14:50:10 -1000 From: "Jack England" Subject: [Histonet] Collagen IHC in mouse tissue--need antibody recommendations! To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Aloha Histonetters, and happy holidays to all! We've been trying to do immunostaining for collagen I on some murine-derived cell lines, with next to no success. I'm almost certain that I've got collagen in there (PSR shows plenty of red, my poor-man's polarizers make the red stuff look birefringent, and SEM images show all kinds of fibrous ECM), but it's not staining for collagen I or III with the antibodies we've got (both from Sigma, produced in mouse ascites, not tested on mouse tissue). I suspect that our antibodies don't react with mouse tissue, but it's possible that the fixation (we use 10% NBF) or the embedding (paraffin, which does involve heating the tissue) could be denaturing the antigenic sites. I've tried enzymatic antigen retrieval, and it didn't work...next step is to get ahold of mouse tails and a cryostat and try for a positive control. My question for anyone that can help is, assuming that the problem is that our antibodies do not react with mouse tissue, can any of you recommend antibodies to collagen I or III that do react with mouse tissue (preferably FFPE mouse tissue, since we don't have a cryostat)? I've checked the U of Iowa hybridoma bank and come up empty, and a Biocompare search gave me one hit--I'm curious if any of you out there can recommend something beyond that one. Any takers? --With warm regards and aloha, Jack England Tissue Genesis, Inc. http://www.tissuegenesis.com _________________________________________________________________ On the road to retirement? Check out MSN Life Events for advice on how to get there! http://lifeevents.msn.com/category.aspx?cid=Retirement ------------------------------ Message: 10 Date: Mon, 13 Dec 2004 20:43:26 -0500 From: John Kiernan Subject: Re: [Histonet] storage of cryosections To: Birthe Schnegelsberg Cc: histonet@pathology.swmed.edu Message-ID: <41BE453E.BA6D31CA@uwo.ca> Content-Type: text/plain; charset=us-ascii Freezing artefacts (ice crystal holes) form while the tissue is being frozen, not during storage of sections. Once the holes are there, because of too-slow freezing, nothing can get rid of them. Drierite (or a similar desiccant) ensures that the stored sections are in a dry atmosphere. You should let the box warm to room temperature before you open it; otherwise water from the air will condense on the cold sections. If they are of unfixed tissue this will cause some damage, which may or may not matter depending on what you intend to do with the slides. John Kiernan London, Canada. ------------------------------------------------ Birthe Schnegelsberg wrote: > > HI. > Do I actually have to add Drierite absorbent into the boxes with the cryo > sections in the -20C freezer to avoid freezing artefacts? > Thanks, birthe > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Tue, 14 Dec 2004 01:52:18 +0000 From: "Christine Tambasco" Subject: RE: [Histonet] IHC ~ Tissue Falling Off Slides To: lpjones@srhs-pa.org, Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format="flowed" I use poly-l-lysine slides for immunos (i make them myself) . Also i use fisher's probe slides and the citra retreival i do in the microwave. I put the slides in a plastic slide mailer with citra - microwave on high for 45sec - 1min then I add more citra and microwave 3 minutes at 50% power. It works for me. Good Luck! Christine Tambasco, HT (ASCP) St. Mary's Hospital , Amsterdam, New York ph-5188417287 >From: "Jones, Laura" >To: "Histonet (E-mail)" >Subject: [Histonet] IHC ~ Tissue Falling Off Slides >Date: Thu, 9 Dec 2004 11:03:38 -0500 > >Hello fellow Histonetters. > >We have suddenly started to have problems with tissue falling off slides >during IHC processing. We suspect the problem may be our slides, although >we have been using the same ones from Mercedes Medical for a while and they >have worked beautifully! We have contacted them, and they are investigating >and very kindly replacing our stock, but in the meantime, our Pathologist >has asked me to ask all of you where else we can find 25 X 75 silinated >slides. We use the Zymed ST-5050 automated stainer, and the 1 X 3 slides >are sometimes a bit to big to fit on the carousel. > >Just for additional info, we seem to be experiencing problems mostly with >the more alkaline retreival solutions - AR-10 (Biogenex) and EDTA (Zymed) >and Trilogy (Cell Marque). We use the Black & Decker steamer, and have >adjusted our times in retrieval from 75 minutes (!) to 20 minutes, per >Pathologist direction. We have also tried 20 minutes heating, then adding >the slides for 20 minutes of boiling, then allowing them to stand for 20-30 >minutes. > >Thanks in advance for all of your knowledge! > >The Histochicks at Sharon Regional > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Tue, 14 Dec 2004 09:28:24 -0500 From: "Pamela Marcum" Subject: [Histonet] Question to assist a someone off this list To: Message-ID: <000e01c4e1e9$2b4a0f80$7f00a8c0@PMARCUM2K> Content-Type: text/plain; charset="iso-8859-1" Good Morning, I have had several request for slides coated with different silanes or charges for people doing cell culture, plastics and some special histology stains. They are looking for negatively charged slides or special coatings. Do any of you on HistoNet know anything about these slides like where I can tell them to go or what type of coatings are available? I am sorry to ask this however, I don't know where else to go and as histologist I am more familiar with the traditional coatings and subbing materials. Thanks for the help or direction! Pamela A Marcum Histology/Microscopy Product Development Manager Polysciences, Inc. 400 Valley Road Warrington, PA 18976 Telephone: 800-523-2575???? Ext. 167 215-343-6484???? Ext. 167 Fax:? 800-343-3291 ???????? 215-343-0214 ------------------------------ Message: 13 Date: Tue, 14 Dec 2004 09:01:13 -0600 From: Philip Oshel Subject: Re: [Histonet] storage of cryosections To: jkiernan@uwo.ca Cc: Histonet@Pathology.swmed.edu Message-ID: Content-Type: text/plain; format=flowed; charset=us-ascii I'm afraid I have to disagree with a portion of John's email: ice crystal damage can happen in storage. Ice crystal growth and refreezing occurs at temperatures above about -60 to -40 deg C, depending on the specifics of the tissue, local environment, and all the rest of the annoying biological realities we deal with. If tissues are frozen rapidly enough to prevent crystal formation, or to form only tiny crystals, the tissues/cells will experience crystal growth if they are warmed above the recrystallization temperature. Much of the damage is actually caused by dehydration as the growing ice crystals pull free water out of cells and intercellular spaces, but as crystal growth continues, it can form holes. The preventative is to store samples in -80 deg C freezers and to warm them too quickly for recrystallizaton and crystal growth to occur. But, there is an even neater trick: store the samples under vacuum in a -80 freezer with *lots* of desiccant (Lots -- no, more ... pile it in the chamber. More!*). Something with a high water capacity: best is 3-4 Angstrom (4 is better than 3, 5 is no good) molecular sieve, next best is silica gel. Leave the samples like this for a few days (hours to weeks, depending on sample size and number -- it's empirical, try it and see), and they will be nicely freeze-dried by vacuum sublimation. They can then be stored at any temperature with desiccant with no worries about ice crystals, or much of anything. On exposure to air, the samples will immediately start rehydrating, so they *must* be at room temperature before removing them from the desiccant chamber, and processed right away. Caveat: I don't think this would adversely affect antigen epitopes, but this should be tested. Phil * If you think you have enough, you don't. >Freezing artefacts (ice crystal holes) form while >the tissue is being frozen, not during storage of >sections. Once the holes are there, because of >too-slow freezing, nothing can get rid of them. > >Drierite (or a similar desiccant) ensures that >the stored sections are in a dry atmosphere. >You should let the box warm to room temperature >before you open it; otherwise water from the air >will condense on the cold sections. If they are of >unfixed tissue this will cause some damage, which may >or may not matter depending on what you intend to >do with the slides. > >John Kiernan >London, Canada. >------------------------------------------------ >Birthe Schnegelsberg wrote: >> >> HI. >> Do I actually have to add Drierite absorbent into the boxes with the cryo >> sections in the -20C freezer to avoid freezing artefacts? >> Thanks, birthe >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) ------------------------------ Message: 14 Date: Tue, 14 Dec 2004 10:44:22 -0500 From: "Bonner, Janet" Subject: RE: [Histonet] Microwave Devices and Proposed Guidelines To: "'Sandy Cheasty'" , "'histonet@pathology.swmed.edu'" Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4127@fh2k093.fhmis.net> Content-Type: text/plain; charset="iso-8859-1" I've found that the issue of commercial vs industrial microwaves boils down to a ventilation problem( possible explosion or respiratory problem on a "stain" becoming an aerosol) as well as QA as far as temperature and time regulation. The microwave should be vented (ours is in the back) to the outside or a fume hood. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sandy Cheasty Sent: Monday, December 13, 2004 11:48 AM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] Microwave Devices and Proposed Guidelines If you receive any info on this would you please let me know. We are currently using a microwave from Wal-Mart. There has got to be some kind of exception somewhere. What about labs that could not justify a lab microwave but has a commercial one that works just as good for special stains only. Thanks, amy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sandy Cheasty Sent: Friday, December 10, 2004 2:58 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] Microwave Devices and Proposed Guidelines I know there are thousands of histology labs that use commercial microwaves safely and have used them safely for years. Does everyone: -violate OSHA? -lie to their lab managers? There must be some way of continuing to use commercial microwave ovens for special stains without being subversive. Don't get me wrong, I usually enjoy being subversive. Can someone please give me a justification for using a commercial microwave or tell me where I can get an OSHA approved one for less than $1000? Thanks again, Elvis -----Original Message----- From: Willis, Donna [mailto:DonnaWillis@texashealth.org] Sent: Friday, December 10, 2004 7:08 AM To: Sandy Cheasty; histonet@pathology.swmed.edu Subject: RE: [Histonet] NCCLS Microwave Device Use Proposed Guideline It is against OSHA guideline (OSHA 29 CFR 1910.303(b)(2))to use a commercial microwave (exp. Wal-Mart). The warranty of commercial units state that the instrument is not to be used for anything except its original use, food preparation. The OSHA guideline would include any equipment in the lab. If someone gets injured the lab would be at fault not the manufacture. Donna Willis Histology Lab Manager Harris Methodist Fort Worth,Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sandy Cheasty Sent: Friday, December 10, 2004 7:40 AM To: histonet@pathology.swmed.edu Subject: [Histonet] NCCLS Microwave Device Use Proposed Guideline I have the GP28-P guideline and read it but am unsure about its recommendations or limitations in using a microwave for special stains only. Is there any consensus among histology labs as to its interpretation? Is it still OK to use a commercial microwave from Wal-Mart without special exhaust system as long as you don't microwave formaldehyde, osmium tetroxide, lead acetate etc. (Paragraph 4.4) and do annual leakage tests? Thanks! Elvis ____________________________________________________________________________ __ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. ____________________________________________________________________________ __ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Note: The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or an agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ____________________________________________________________________________ __ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. Sign up for your free MemorialCare Medical Information and Access Card at http://www.memorialcare.com/apps/AccessCard/AboutCard.cfm _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. ------------------------------ Message: 15 Date: Tue, 14 Dec 2004 10:56:25 -0500 From: Rita Riddle Subject: [Histonet] unscribe To: "'Histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. ------------------------------ Message: 16 Date: Tue, 14 Dec 2004 09:04:37 -0700 From: Gayle Callis Subject: Re: [Histonet] storage of cryosections To: "Birthe Schnegelsberg" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041214084115.01b0b540@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Birthe, Initially, how you snap freeze is the first defense against freezing artefacts. Frozen section stored with a dessicant is a good idea keep the sections dry during storage. The storage container must stay closed to equilibrate FS to room temperature. Water condensation is the enemy. Be sure to air dry the sections before going into storage also. A black plastic 25 slide capacity box works well. Also, opening a box directly from freezer and taking a few sections out, then returning unused sections to freezer must be avoided due FS freeze thaw. Put as many slides as you need per storage container for a staining session. Drierite is anhydrous calcium chloride and has a tendency to exfoliate fine CaCl2dust all over everything including your sections which bothered me. Try 10 to 18 mesh silica gel, (Fisher# S161-500). The little beads have blue indicator so you know when dessicant is depleted. Put Silica gel in a nylon processing bags (Thermo Electron or StatLab) or embedding bags aka tea bags from Fisher, fold over top, and staple on fold the keep bags closed. Temperature is critical for storage - if you have -80C available, use that instead of -20C, and NEVER store sections in a self defrosting freezer found in a refrigerator, freeze dry cycle is very damaging. Hopefully you can store FS at temperature lower than -20C to preserve antigenicity over a long period of time. It may work for short term storage and robust antigens. At 02:23 PM 12/13/2004, you wrote: >HI. >Do I actually have to add Drierite absorbent into the boxes with the cryo >sections in the -20C freezer to avoid freezing artefacts? >Thanks, birthe > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 17 Date: Tue, 14 Dec 2004 09:08:09 -0700 From: "Brianna Jackson" Subject: [Histonet] Payscale To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hello, Can histotechs working in labs with separate IHC and Histo departments tell me if you have the same payscale? I've been told that IHC techs (even HTL certified) cannot be paid the same as production bench techs. I was just wondering if this is how it works in other labs. Thanks for your time, Brianna Jackson, BS, QIHC Denver, CO 303-512-2220 ------------------------------ Message: 18 Date: Tue, 14 Dec 2004 11:09:48 -0500 From: "Vicki Gauch" Subject: [Histonet] Position To: Message-ID: Content-Type: text/plain; charset=US-ASCII I have a tech with 10 years of experience who will be relocating to the Scottsdale,Arizona region who is seeking a Histotech position in that area. Does anyone know of any openings? Please send any correspondence to this e-mail address as she is not currently able to access the Histonet. Thank you for your help. Vicki Gauch Albany Medical Center Albany, NY ------------------------------ Message: 19 Date: Tue, 14 Dec 2004 09:20:04 -0700 From: Gayle Callis Subject: [Histonet] Special coatings for slides To: "Pamela Marcum" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20041214091203.01b4e368@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Pam, I believe Erie Scientific is doing some of this from what my Erie rep tells me. Erie website has 800 number for tech services. They have some new things for Microarray work. Also try Grace Bio-Labs, they have a website, and you can call them for detailed discussion. Be prepared for some pricey items. At 07:28 AM 12/14/2004, you wrote: >Good Morning, > >I have had several request for slides coated with different silanes or >charges for people doing cell culture, plastics and some special histology >stains. They are looking for negatively charged slides or special coatings. >Do any of you on HistoNet know anything about these slides like where I can >tell them to go or what type of coatings are available? > >I am sorry to ask this however, I don't know where else to go and as >histologist I am more familiar with the traditional coatings and subbing >materials. > >Thanks for the help or direction! > >Pamela A Marcum >Histology/Microscopy >Product Development Manager >Polysciences, Inc. >400 Valley Road >Warrington, PA 18976 >Telephone: 800-523-2575 Ext. 167 > 215-343-6484 Ext. 167 > >Fax: 800-343-3291 > 215-343-0214 > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 20 Date: Tue, 14 Dec 2004 10:21:33 -0600 From: "Cory Collins" Subject: [Histonet] Varicella-zoster To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Howdy!! I work for a reference lab in Dallas and we are having trouble finding control tissue for Varicella-zoster. Does anyone know of a vendor that sells control slides for Varcilla-zoster? Or maybe know someone that has some extra paraffin embedded tissue that is positive for Varicella-zoster. Thanks, Cory Cory Collins, HT (ASCP) QIHC Immunohistochemistry Supervisor ProPath 8267 Elmbrook Drive Suite 100 Dallas, TX 75247 214-638-2000 ext 2027 214-237-1730 Fax To learn more about ProPath, please visit http://www.ProPathLab.com ______________________________________________________________________________ This e-mail may contain confidential or privileged information. If you think you have received this e-mail in error, please advise the sender by reply e-mail and then delete this e-mail immediately. ------------------------------ Message: 21 Date: Tue, 14 Dec 2004 09:19:47 -0800 From: "Mark Elliott" Subject: [Histonet] MMP-1 in Bouin's fixed tissue To: Message-ID: Content-Type: text/plain; charset=US-ASCII Has anyone had any experience staining for MMP-1 in Bouin's fixed human tissues? One of the Hospital's we collaborate with fixes their lungs in Bouin's. We have tried staining with no antigen retrieval and with/without overnight incubation in primary. Have also tried antigen retrieval using citra pH 6 in 95 C water bath and in autoclave with no luck. We are using MAB 1346 from Chemicon followed by APAAP technique. Are going to try high pH antigen retrieval and possibly trypsin but would appreciate any other suggestions. Thanks Mark Elliott iCAPTURE Centre Vancouver BC ------------------------------ Message: 22 Date: Tue, 14 Dec 2004 09:34:31 -0800 (PST) From: Paula Pierce Subject: [Histonet] Collagen IHC in mouse tissue - need antibody recommendations To: histonet@lists.utsouthwestern.edu Message-ID: <20041214173431.64936.qmail@web50308.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Have you tried just a histochemical stain for collagen such as Masson's Trichrome? Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com ------------------------------ Message: 23 Date: Tue, 14 Dec 2004 17:38:18 -0000 From: "Marshall Terry Dr, Consultant Histopathologist" Subject: RE: [Histonet] Collagen IHC in mouse tissue - need antibodyrecommendations To: "Paula Pierce" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Masson - histochemical? 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: Paula Pierce [mailto:contact@excaliburpathology.com] Sent: 14 December 2004 17:35 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Collagen IHC in mouse tissue - need antibodyrecommendations Have you tried just a histochemical stain for collagen such as Masson's Trichrome? Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 13, Issue 20 **************************************** From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 15 02:45:56 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Ventana Benchmark[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5F5@bhrv-nt-11.bhrv.nwest.nhs.uk> I look forward to the replies for this Cindy as I wasn't aware you could do HPV on ThinPrep's on this. Can you do it on the Dako too. We are moving to the LBC platform this Spring and the ability to 'do' HPV may be an advantage; however we are using the SurePath technology is that a problem? -----Original Message----- From: Cindy DuBois [mailto:dpahisto@yahoo.com] Sent: 14 December 2004 18:10 To: Histonet Subject: [Histonet] Ventana Benchmark[Scanned] Our lab is considering purchasing a Ventana Benchmark to do both immuno's and HPV testing (on thin preps). Has anyone had difficulty with this machine? With our volume we would have to do 2 runs a day, one for immunos - and one for HPV. I was also told we would begin doing ER/PR also. Where can I get the most valuable information regaring ER/PR staining? Are there any classes available? Cindy DuBois, HT ASCP DELTA PATHOLOGY ASSOC. STOCKTON CA --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com/a _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 15 02:56:00 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] recycled alcohol[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B5FA@bhrv-nt-11.bhrv.nwest.nhs.uk> I've some 'recycled alcohol' it's a funny colour, it's yellow. -----Original Message----- From: Laurie Colbert [mailto:laurie.colbert@huntingtonhospital.com] Sent: 14 December 2004 20:03 To: Christine Tambasco; Histonet (E-mail) Subject: RE: [Histonet] recycled alcohol[Scanned] Do you flush the tank with alcohol when you switch from Clear Rite to alcohol? Laurie Colbert -----Original Message----- From: Christine Tambasco [mailto:immrstambo@hotmail.com] Sent: Friday, December 10, 2004 5:58 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] recycled alcohol Does anyone have any experience with the CBG recycler and the Leica tissue processor? My recycled alcohol seems to have clear rite in it. The rep says it could be fat from the types of tissue processed and not clear rite at all. The recycled alcohol turns cloudly when mixed with water. I am relatively sure there isnt any clear rite in the alcohol before recycling, but I tested the alcohol before and after, and i seems to always be contaminated. I should think it couldnt be my processor (at least hope not) Thanks for any input. Christine Tambasco, HT (ASCP) St. Mary's Hospital Lab Amsterdam, New York _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 15 05:35:15 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Ultrasound directed FNAC of gastric lesion, lymph nodes, etc[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3ED82@bhrv-nt-11.bhrv.nwest.nhs.uk> I know this is a Cytology-type question, but CytopathNet seems to have died a death. I posted a question to it the other day and got answers from second hand car dealers, most bizarre. I have been asked by my Pathologists and Gastro Surgeons to set up a diagnostic FNAC service for 'gut' cancer by using this new Ultrasound equipment; the sender is on the tip of the flexible scope. You can see CT like clarity, really rather kewl. I'm interested in providing a service whereby 'adequacy' is gauged at the 'bedside' and expanding ICC on ThinPreps to aid diagnosis. I have experience in providing this service for CT orientated FNAC but without the ICC involvement. Anyone any experience, I think the Gastro man would like to link up with anyone providing this service due to his relative inexperience. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 From Julie.Sanders <@t> med.va.gov Wed Dec 15 07:18:26 2004 From: Julie.Sanders <@t> med.va.gov (Julie.Sanders@med.va.gov) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] RE: Histonet Digest, Vol 13, Issue 21 Message-ID: <457381D92B01BD44B21CF37CC02EBDFD02927539@vhacinexc2.v10.med.va.gov> Cindy, We have the Benchmark LT (which means it only holds 20 slides as opposed the XT which holds 30). We do all of our immunos during the day and HPV's at night. However, if there is enough room, we run them at the same time. It really has been a time saver. Since I am at a VA we rarely have ER/PR so I can't help you on that. However, I have found the Benchmark to be a great instrument and we've had no trouble with it. Julie Sanders, BA, HT(ASCP) Supervisor, Anatomic Pathology VA Medical Center, Cincinnati, Oh. Message: 1 Date: Tue, 14 Dec 2004 10:09:38 -0800 (PST) From: Cindy DuBois Subject: [Histonet] Ventana Benchmark To: Histonet Message-ID: <20041214180938.5019.qmail@web41305.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Our lab is considering purchasing a Ventana Benchmark to do both immuno's and HPV testing (on thin preps). Has anyone had difficulty with this machine? With our volume we would have to do 2 runs a day, one for immunos - and one for HPV. I was also told we would begin doing ER/PR also. Where can I get the most valuable information regaring ER/PR staining? Are there any classes available? Cindy DuBois, HT ASCP DELTA PATHOLOGY ASSOC. STOCKTON CA --------------------------------- From dsnider <@t> shrinenet.org Wed Dec 15 06:17:36 2004 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] Training program for processing GMA specimens Message-ID: Does anyone on the list know of any professional training in this area? I have recently accepted a position which requires me to know this. While working through the existing backlog of work, I taught myself to cut them, but in 15 years of experience I have never been exposed this procedure. Any help will be greatly appreciated. Thanks in advance... Deanna Snider HT ASCP Shriners Hospital - Cincinnati, OH CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From wasielewski.reinhard.von <@t> mh-hannover.de Wed Dec 15 08:49:37 2004 From: wasielewski.reinhard.von <@t> mh-hannover.de (wasielewski.reinhard.von@mh-hannover.de) Date: Fri Sep 16 15:24:24 2005 Subject: [Histonet] CD34 and CD31 in paraffin mouse tissue Message-ID: <41C05D11.15565.2FFF9314@localhost> Hello Histonetters, we are looking to immunostain endothelial cells (vessels) in FFPE mouse tissue. There is no cross-reactivity to the human epitopes, therefore the antibodies for human tissue don't work (we have tried it). Is there anyone out there who can help us ? (Clones, suppliers, ...) Answers wanted :-) Best wishes for the rest of this year Reinhard von Wasielewski PD Dr. med. Reinhard von Wasielewski From liz <@t> premierlab.com Wed Dec 15 09:51:34 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] CD34 and CD31 in paraffin mouse tissue In-Reply-To: <41C05D11.15565.2FFF9314@localhost> Message-ID: <000201c4e2bd$f47c2710$76d48a80@AMY> Reinhard Santa Cruz has a goat polyclonal for CD-31 (PECAM-1) that will work in FFPE tissue with antigen retrieval. Cat SC-1056. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of wasielewski.reinhard.von@mh-hannover.de Sent: Wednesday, December 15, 2004 7:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD34 and CD31 in paraffin mouse tissue Hello Histonetters, we are looking to immunostain endothelial cells (vessels) in FFPE mouse tissue. There is no cross-reactivity to the human epitopes, therefore the antibodies for human tissue don't work (we have tried it). Is there anyone out there who can help us ? (Clones, suppliers, ...) Answers wanted :-) Best wishes for the rest of this year Reinhard von Wasielewski PD Dr. med. Reinhard von Wasielewski _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jhnspam <@t> aol.com Wed Dec 15 10:57:04 2004 From: jhnspam <@t> aol.com (jhnspam@aol.com) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Temporary Services Message-ID: <24.66633132.2ef1c6e0@aol.com> I work for a lab that does 61,000 surgicals a year. I am thinking about contacting a temporary service to fill vacant positions. Can anyone give me a ball park figure of what I should expect to pay for this service? Jennifer Brasell From peoshel <@t> wisc.edu Wed Dec 15 11:28:58 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] (no subject) Message-ID: Charles, A couple of points, mostly for the fun of the discussion: There is another form of frozen water, in the temperature/pressure realms biologists are concerned with: evanescent spherulites. Some people argue that vitreous water is never obtained, except in *very* small volumes (microliters) at *very* high freezing rates (>10,000 deg C/sec), but rather that evanescent spherulites are formed. Practically speaking, the results are the same. I agree about the cubic ice. I think cubic ice is formed during freezing with a high-pressure freezing (HPF) apparatus, but that's speculation from pressure/temperature tables. I haven't meant anyone who really knows what's happening during high-pressure freezing. (Most are surprised when it's mentioned that a pressure spike necessarily means a temperature spike before the cooling.) There is one point in your note with which I disagree: "...and some ice crystals will form in the interior of any piece of tissue over 10mm from the cold source." Under ideal conditions, with the best freezing method -- HPF or rapid plunging into slush nitrogen -- crystal-free freezing will occur only to a depth of ~500 nm at best. It might perhaps be possible to get freezing rapid enough that crystals are not easily seen by light microscopy at greater distances from the freezing surface, but there will definitely be crystals and freezing artifacts 10 mm from the surface of the specimen. Phil >I agree that initial freezing is most important, but that freezing >artifact can develop in storage because the ice will reform and >crystalize. See the following link for a thorough discussion. > >http://www.myneurolab.com/global/Manuals/Tips%20and%20Techniques%20Freezing%20Artifact.pdf > >Drying agents are thus a good idea. > > > > >Cordially, >Charles W. Scouten, Ph.D. >myNeuroLab.com >5918 Evergreen Blvd. >St. Louis, MO 63134 >Ph: 314 522 0300 >FAX 314 522 0377 >cwscouten@myneurolab.com >http://www.myneurolab.com -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From peoshel <@t> wisc.edu Wed Dec 15 11:32:30 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] storage of cryosections Message-ID: Charles, A couple of points, mostly for the fun of the discussion: There is another form of frozen water, in the temperature/pressure realms biologists are concerned with: evanescent spherulites. Some people argue that vitreous water is never obtained, except in *very* small volumes (microliters) at *very* high freezing rates (>10,000 deg C/sec), but rather that evanescent spherulites are formed. Practically speaking, the results are the same. I agree about the cubic ice. I think cubic ice is formed during freezing with a high-pressure freezing (HPF) apparatus, but that's speculation from pressure/temperature tables. I haven't meant anyone who really knows what's happening during high-pressure freezing. (Most are surprised when it's mentioned that a pressure spike necessarily means a temperature spike before the cooling.) There is one point in your note with which I disagree: "...and some ice crystals will form in the interior of any piece of tissue over 10mm from the cold source." Under ideal conditions, with the best freezing method -- HPF or rapid plunging into slush nitrogen -- crystal-free freezing will occur only to a depth of ~500 nm at best. It might perhaps be possible to get freezing rapid enough that crystals are not easily seen by light microscopy at greater distances from the freezing surface, but there will definitely be crystals and freezing artifacts 10 mm from the surface of the specimen. Phil >I agree that initial freezing is most important, but that freezing >artifact can develop in storage because the ice will reform and >crystalize. See the following link for a thorough discussion. > >http://www.myneurolab.com/global/Manuals/Tips%20and%20Techniques%20Freezing%20Artifact.pdf > >Drying agents are thus a good idea. > > > > >Cordially, >Charles W. Scouten, Ph.D. >myNeuroLab.com >5918 Evergreen Blvd. >St. Louis, MO 63134 >Ph: 314 522 0300 FAX 314 522 0377 >cwscouten@myneurolab.com >http://www.myneurolab.com -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From vazquezr <@t> ohsu.edu Wed Dec 15 11:46:14 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Temporary Services Message-ID: Jennifer where are you located? Robyn OHSU >>> 12/15/2004 8:57:04 AM >>> I work for a lab that does 61,000 surgicals a year. I am thinking about contacting a temporary service to fill vacant positions. Can anyone give me a ball park figure of what I should expect to pay for this service? Jennifer Brasell _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jhnspam <@t> aol.com Wed Dec 15 12:10:50 2004 From: jhnspam <@t> aol.com (jhnspam@aol.com) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Temporary Services Message-ID: <7f.5370d324.2ef1d82a@aol.com> Tennessee From MichelM9 <@t> chw.edu.au Wed Dec 15 15:06:28 2004 From: MichelM9 <@t> chw.edu.au (Michelle McDonald) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] CD34 and CD31 in paraffin mouse tissue Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740C952B21@simba.kids> Dear Liz, Would you know whether this antibody from Santa Cruz would work on rat tissue. I am currently trying to get a CD31 antibody from Calbiochem to work on FFPE rat bone sections (decalcified) and I am having no luck. Can you suggest an antibody I could try or recommend an antigen retrieval technique. I have tried Proteinase K and Pepsin. Thank you in advance for your advice Michelle Michelle McDonald B.MedSci PhD Student Research Assistant Orthopaedic Research Dept. The Children's Hospital Westmead. Westmead NSW 2145 Australia Ph. +612 9845 3087 Fax. +612 9845 3078 -----Original Message----- From: Elizabeth Chlipala [mailto:liz@premierlab.com] Sent: Thursday, 16 December 2004 2:52 AM To: wasielewski.reinhard.von@mh-hannover.de; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] CD34 and CD31 in paraffin mouse tissue Reinhard Santa Cruz has a goat polyclonal for CD-31 (PECAM-1) that will work in FFPE tissue with antigen retrieval. Cat SC-1056. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of wasielewski.reinhard.von@mh-hannover.de Sent: Wednesday, December 15, 2004 7:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD34 and CD31 in paraffin mouse tissue Hello Histonetters, we are looking to immunostain endothelial cells (vessels) in FFPE mouse tissue. There is no cross-reactivity to the human epitopes, therefore the antibodies for human tissue don't work (we have tried it). Is there anyone out there who can help us ? (Clones, suppliers, ...) Answers wanted :-) Best wishes for the rest of this year Reinhard von Wasielewski PD Dr. med. Reinhard von Wasielewski _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From JCarpenter764 <@t> aol.com Wed Dec 15 15:13:12 2004 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] certified histo pay Message-ID: <12f.528f6788.2ef202e8@aol.com> hey everyone!!! Here is a situation...If I have been employed with the same facility for almost 5 years and in histology for over two years full time (that is what i needed to take the histology exam) and had over two years full time experience in April of last year and now it is December what is the going pay rate that I should be receiving after my certification in histology as an HT? Just curious...thanks Jennell From portera203 <@t> yahoo.com Wed Dec 15 15:17:02 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Looking for direct email address for Dr. Van der Loos Message-ID: <20041215211702.93957.qmail@web40905.mail.yahoo.com> Would like to correspond with Dr. Van der Loos about problems we are having in our laboratory with double staining. I posted an enquiry several weeks ago and have had more than one reference that he is the person to talk with about this. I would greatly appreciate his input about the process we are trying to utilize. Amy S.Porter, HT(ASCP) Michigan State University Department of Physiology Division of Human Pathology College of Human Medicine portera203@yahoo.com --------------------------------- Do you Yahoo!? Send holiday email and support a worthy cause. Do good. From claudiamel2000 <@t> yahoo.com Wed Dec 15 15:17:27 2004 From: claudiamel2000 <@t> yahoo.com (claudia melidona) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] IHC for Prostate carcinoma Message-ID: <20041215211727.60874.qmail@web60804.mail.yahoo.com> Need some info on CK903/P63/Racemase staining protocol on single slide. Hope there's some help out there in histoland! --------------------------------- Do you Yahoo!? The all-new My Yahoo! – Get yours free! From liz <@t> premierlab.com Wed Dec 15 15:20:57 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] CD34 and CD31 in paraffin mouse tissue In-Reply-To: <1CF2E2E5BB36D5119E7A0008C791F3740C952B21@simba.kids> Message-ID: <000001c4e2eb$f82ac140$76d48a80@AMY> Michelle It also works on rat. I have tried it on formalin perfused rat brains and it works. I use steam retrieval for 20 minutes, antibody at 1:500, 30 minutes RT, a rabbit anti-goat from vector at 1:400 and then rabbit envision from Dako to amplify. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: Michelle McDonald [mailto:MichelM9@chw.edu.au] Sent: Wednesday, December 15, 2004 2:06 PM To: 'Elizabeth Chlipala'; wasielewski.reinhard.von@mh-hannover.de; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] CD34 and CD31 in paraffin mouse tissue Dear Liz, Would you know whether this antibody from Santa Cruz would work on rat tissue. I am currently trying to get a CD31 antibody from Calbiochem to work on FFPE rat bone sections (decalcified) and I am having no luck. Can you suggest an antibody I could try or recommend an antigen retrieval technique. I have tried Proteinase K and Pepsin. Thank you in advance for your advice Michelle Michelle McDonald B.MedSci PhD Student Research Assistant Orthopaedic Research Dept. The Children's Hospital Westmead. Westmead NSW 2145 Australia Ph. +612 9845 3087 Fax. +612 9845 3078 -----Original Message----- From: Elizabeth Chlipala [mailto:liz@premierlab.com] Sent: Thursday, 16 December 2004 2:52 AM To: wasielewski.reinhard.von@mh-hannover.de; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] CD34 and CD31 in paraffin mouse tissue Reinhard Santa Cruz has a goat polyclonal for CD-31 (PECAM-1) that will work in FFPE tissue with antigen retrieval. Cat SC-1056. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of wasielewski.reinhard.von@mh-hannover.de Sent: Wednesday, December 15, 2004 7:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD34 and CD31 in paraffin mouse tissue Hello Histonetters, we are looking to immunostain endothelial cells (vessels) in FFPE mouse tissue. There is no cross-reactivity to the human epitopes, therefore the antibodies for human tissue don't work (we have tried it). Is there anyone out there who can help us ? (Clones, suppliers, ...) Answers wanted :-) Best wishes for the rest of this year Reinhard von Wasielewski PD Dr. med. Reinhard von Wasielewski _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From gcallis <@t> montana.edu Wed Dec 15 15:52:21 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Looking for direct email address for Dr. Van der Loos In-Reply-To: <20041215211702.93957.qmail@web40905.mail.yahoo.com> References: <20041215211702.93957.qmail@web40905.mail.yahoo.com> Message-ID: <6.0.0.22.1.20041215145047.01b3d028@gemini.msu.montana.edu> AMy, Chris monitors Histonet, but I forwarded your message to him. He will respond to you. Just in case, "C.M. vander Loos" At 02:17 PM 12/15/2004, you wrote: >Would like to correspond with Dr. Van der Loos about problems we are >having in our laboratory with double staining. I posted an enquiry >several weeks ago and have had more than one reference that he is the >person to talk with about this. I would greatly appreciate his input >about the process we are trying to utilize. > > >Amy S.Porter, HT(ASCP) >Michigan State University >Department of Physiology >Division of Human Pathology >College of Human Medicine >portera203@yahoo.com > > > >--------------------------------- >Do you Yahoo!? > Send holiday email and support a worthy cause. Do good. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From tahseen <@t> brain.net.pk Sun Dec 12 14:07:15 2004 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Protocol for grossing the amputation. Message-ID: <000201c4e2e0$525d1480$972bfea9@m7c0y4> Can anyone help me with a protocol for grossing the amputation.I'm particularly interested in looking mug saw. Tahseen. From Jason.PALMER <@t> svhm.org.au Wed Dec 15 20:31:23 2004 From: Jason.PALMER <@t> svhm.org.au (PALMER Jason (SVHM)) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] mouse endothelial cell staining Message-ID: We use a rat anti mouse CD31 from BD Biosciences that works nicely at 1:50 - 1:100 with 8 minutes proteinase K digestion. Cheers, Jason Jason Palmer Bernard O'Brien Institute of Microsurgery 42 Fitzroy St, Fitzroy Victoria 3065 Australia tel +61 3 9288 4018 fax +61 3 9416 0926 email: palmerj@svhm.org.au Date: Wed, 15 Dec 2004 15:49:37 +0100 From: wasielewski.reinhard.von@mh-hannover.de Subject: [Histonet] CD34 and CD31 in paraffin mouse tissue To: histonet@lists.utsouthwestern.edu Message-ID: <41C05D11.15565.2FFF9314@localhost> Content-Type: text/plain; charset=US-ASCII Hello Histonetters, we are looking to immunostain endothelial cells (vessels) in FFPE mouse tissue. There is no cross-reactivity to the human epitopes, therefore the antibodies for human tissue don't work (we have tried it). Is there anyone out there who can help us ? (Clones, suppliers, ...) Answers wanted :-) Best wishes for the rest of this year Reinhard von Wasielewski PD Dr. med. Reinhard von Wasielewski -------------- next part -------------- Disclaimer : The contents of this e-mail including any attachments are intended only for the person or entity to which this e-mail is addressed and may contain confidential, privileged and/or commercially sensitive material. If you are not, or believe you may not be, the intended recipient, please advise the sender immediately by return e-mail, delete this e-mail and destroy any copies. From TMcNemar <@t> lmhealth.org Thu Dec 16 08:44:22 2004 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] certified histo pay Message-ID: <6CD94D97ED7D924BA5C2B588FA952821396762@nt_exchange.lmhealth.org> Unfortunately, we do not pay anything for passing the exam. It is considered (by those above me) just a normal requirement of the job. It is expected. I don't know about elsewhere... Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Wednesday, December 15, 2004 4:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] certified histo pay hey everyone!!! Here is a situation...If I have been employed with the same facility for almost 5 years and in histology for over two years full time (that is what i needed to take the histology exam) and had over two years full time experience in April of last year and now it is December what is the going pay rate that I should be receiving after my certification in histology as an HT? Just curious...thanks Jennell _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Thu Dec 16 09:11:24 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] storage of cryosections Message-ID: My experience is all with freezing whole rodent brains. I have seen severe swiss cheese artifact, mild freezing arifact, and tissue that looked all good to me under light microscopy. I don't know what happens at EM levels, or with other tissues, but it is possible to immersion freeze (or powedered dry ice) whole rodent brain, approximately 1 cubic centimeter (thinner in one dimension, larger in the other two), examine under light microscopy, and see no detectable freezing artifact. I have tried, but not yet found any reference to the evanescent spherulites as water, only other polymers, on the web. The refereence I sited in my linked article did not mention such a structure. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Philip Oshel Sent: Wednesday, December 15, 2004 11:33 AM To: Histonet@Pathology.swmed.edu Subject: RE: [Histonet] storage of cryosections Charles, A couple of points, mostly for the fun of the discussion: There is another form of frozen water, in the temperature/pressure realms biologists are concerned with: evanescent spherulites. Some people argue that vitreous water is never obtained, except in *very* small volumes (microliters) at *very* high freezing rates (>10,000 deg C/sec), but rather that evanescent spherulites are formed. Practically speaking, the results are the same. I agree about the cubic ice. I think cubic ice is formed during freezing with a high-pressure freezing (HPF) apparatus, but that's speculation from pressure/temperature tables. I haven't meant anyone who really knows what's happening during high-pressure freezing. (Most are surprised when it's mentioned that a pressure spike necessarily means a temperature spike before the cooling.) There is one point in your note with which I disagree: "...and some ice crystals will form in the interior of any piece of tissue over 10mm from the cold source." Under ideal conditions, with the best freezing method -- HPF or rapid plunging into slush nitrogen -- crystal-free freezing will occur only to a depth of ~500 nm at best. It might perhaps be possible to get freezing rapid enough that crystals are not easily seen by light microscopy at greater distances from the freezing surface, but there will definitely be crystals and freezing artifacts 10 mm from the surface of the specimen. Phil >I agree that initial freezing is most important, but that freezing >artifact can develop in storage because the ice will reform and >crystalize. See the following link for a thorough discussion. > >http://www.myneurolab.com/global/Manuals/Tips%20and%20Techniques%20Free >zing%20Artifact.pdf > >Drying agents are thus a good idea. > > > > >Cordially, >Charles W. Scouten, Ph.D. >myNeuroLab.com >5918 Evergreen Blvd. >St. Louis, MO 63134 >Ph: 314 522 0300 FAX 314 522 0377 >cwscouten@myneurolab.com >http://www.myneurolab.com -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Thu Dec 16 10:08:00 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] storage of cryosections Message-ID: Once the tissue is well fixed (24 hours in fixative?), storage in PBS or in Fixative is not critical. Storage in sucrose until it sinks will ensure that freezing rate is not as critical, since smaller and fewer crystals will form in the disruptive sucrose environment. Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com ________________________________ From: Frouwke Kuijpers [mailto:F.Kuijpers@science.ru.nl] Sent: Thursday, December 16, 2004 8:00 AM To: Charles Scouten Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] storage of cryosections Charles, a question: When the tissue is emersed in sucrose 30% is the storage than less critical because there are hardly any cristals formed? Frouwke F.J.Kuijpers-Kwant Dept. Cellular Animal Physiology University of Nijmegen Toernooiveld 1 6525 ED Nijmegen frouwke@sci.kun.nl From ddeibler <@t> centexpathlab.com Thu Dec 16 10:19:49 2004 From: ddeibler <@t> centexpathlab.com (David Deibler) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] HT Position Message-ID: <000801c4e38b$119b0d70$2f01010a@tamtron.ctpl.dns> HT position opening : Position opening at Central Texas Pathology Laboratory located in Waco,Texas. Private lab owned by 10 pathologists seeking an HT for immediate employment. Must be ASCP registered. Must have 3-5 years post certification employment experience in Histology laboratory. Must be proficient at embedding, sectioning and special stains. Frozen section experience is also a must. Our pathologists are a great group to work for and we have excellent benefits. Contact: David Deibler (Histology Supervisor) 254-752-9621 ext. 213 or ext 256 (leave message) ddeibler@centexpathlab.com From lloyd.3 <@t> osu.edu Thu Dec 16 10:25:29 2004 From: lloyd.3 <@t> osu.edu (Mary Lloyd) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] rabbit bone specimens Message-ID: <5.1.0.14.2.20041216110632.00a7be60@pop.service.ohio-state.edu> I am working on a research projects with rabbit femurs and tibias. They are decalcified in cal-ex from fisher for 2 weeks and then processed for 1.5 hours in each solution on the processor. 2 changes of 70, 95, 100%, xylene and then paraffin. I am really having problems with the cartilage folding right at the curve of the specimen. It seems as though the bone just below the cartilage is not thoroughly processed or hard. When I do feel I get a decent section it seems to fold when I stain. I am using plus slides. I don't normally do a lot of research so if anyone can help me with this problem either with cutting or processing solutions I would really appreciate the help. From Charles.Embrey <@t> carle.com Thu Dec 16 10:35:55 2004 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Possible PA Position Message-ID: I am sending this out to test the waters. My pathologists met yesterday and felt it may be time to consider adding a second Pathologists' Assistant. Must be ASCP certified as a Pathologists' Assistant or eligible. If anyone is interested in coming to central Illinois send me an e-mail or give me a call at 217-383-3666. I am at Carle Clinic Association in Champaign/Urbana, home of the University of Illinois "Fighting Illini". Charles Embrey Pathologists' Assistant / Histology MGR Carle Clinic, Urbana IL. From sladd <@t> hsc.usf.edu Thu Dec 16 10:37:13 2004 From: sladd <@t> hsc.usf.edu (S Ladd) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] certified histo pay In-Reply-To: <6CD94D97ED7D924BA5C2B588FA952821396762@nt_exchange.lmhealth.org> Message-ID: ...and in Florida we can't get a license without ASCP certification... -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Tom McNemar Sent: Thursday, December 16, 2004 9:44 AM To: 'JCarpenter764@aol.com'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] certified histo pay Unfortunately, we do not pay anything for passing the exam. It is considered (by those above me) just a normal requirement of the job. It is expected. I don't know about elsewhere... Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: JCarpenter764@aol.com [mailto:JCarpenter764@aol.com] Sent: Wednesday, December 15, 2004 4:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] certified histo pay hey everyone!!! Here is a situation...If I have been employed with the same facility for almost 5 years and in histology for over two years full time (that is what i needed to take the histology exam) and had over two years full time experience in April of last year and now it is December what is the going pay rate that I should be receiving after my certification in histology as an HT? Just curious...thanks Jennell _______________________________________________ Histonet mailing list Histonet@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 liz <@t> premierlab.com Thu Dec 16 10:44:23 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] rabbit bone specimens In-Reply-To: <5.1.0.14.2.20041216110632.00a7be60@pop.service.ohio-state.edu> Message-ID: <000201c4e38e$7fb70ae0$76d48a80@AMY> Mary If the blocks are not decaled properly then it might help to soak the blocks for about 20 minutes in decal solution and place them back on ice and then cut them. When we section any joints, after sectioning we place the slides flat on a hot plate overnight. The hot plate needs to be around 45?C, warm enough for the sections to flatten out, but not too hot that the paraffin melts. If they are too hot the articular cartilage will curl. One other thing, you need to let the slides drain a bit, any water under the section when you lay it flat on the hot plate is going to cause problems, but you can?t let them drain too much so that the sections dry out and turn white. If you are staining with H&E you might still get flipping of the articular cartilage, using a new sharp knife can help with this. If you are staining with toluidine blue, this is what you can do if your sections are fipping. After the TB, rinse in water, examine your slides individually for flipping and you can flip the cartilage back by dipping the section gently in water to get the cartilage to stay flat, then lay your slides flat on the hot plate to dry and then coverslip from xylene. Good luck. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mary Lloyd Sent: Thursday, December 16, 2004 9:25 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] rabbit bone specimens I am working on a research projects with rabbit femurs and tibias. They are decalcified in cal-ex from fisher for 2 weeks and then processed for 1.5 hours in each solution on the processor. 2 changes of 70, 95, 100%, xylene and then paraffin. I am really having problems with the cartilage folding right at the curve of the specimen. It seems as though the bone just below the cartilage is not thoroughly processed or hard. When I do feel I get a decent section it seems to fold when I stain. I am using plus slides. I don't normally do a lot of research so if anyone can help me with this problem either with cutting or processing solutions I would really appreciate the help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Thu Dec 16 10:48:00 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] rabbit bone specimens[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3ED9F@bhrv-nt-11.bhrv.nwest.nhs.uk> I would have used celloidin and low viscosity nitrocellulose embedding for such things as eyes and bones. But I'm sure this 'old' technique has been superseded by a posher new technic. If not, Lillie has a good section on it on page 92 "Histopathologic Technic and Practical Histochemistery". If you can't find it or are unable to find anything better then send me an e-mail. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Mary Lloyd [mailto:lloyd.3@osu.edu] Sent: 16 December 2004 16:25 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] rabbit bone specimens[Scanned] I am working on a research projects with rabbit femurs and tibias. They are decalcified in cal-ex from fisher for 2 weeks and then processed for 1.5 hours in each solution on the processor. 2 changes of 70, 95, 100%, xylene and then paraffin. I am really having problems with the cartilage folding right at the curve of the specimen. It seems as though the bone just below the cartilage is not thoroughly processed or hard. When I do feel I get a decent section it seems to fold when I stain. I am using plus slides. I don't normally do a lot of research so if anyone can help me with this problem either with cutting or processing solutions I would really appreciate the help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mhorne <@t> upei.ca Thu Dec 16 10:31:54 2004 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] mouse endothelial cell staining In-Reply-To: Message-ID: <41C18E49.14815.296452@localhost> Hello Histonetters, I am looking at doing IHC on rat endothelial cells ( aortas) in Epon. Had been looking into using von Willebrand Factor ( Factor VIII ). I noticed in this Thread that people are using CD31. Curious as to the pros and cons of CD31 vs v WF. Has anyone out there worked with an endothelial cell marker in Epon? Any info much appreciated, 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 Jamie.Dukes <@t> se.amedd.army.mil Thu Dec 16 11:55:47 2004 From: Jamie.Dukes <@t> se.amedd.army.mil (Dukes, Jamie Mr EAMC) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] AUTOSTAINER Message-ID: HELLO THIS IS JAMIE FROM FORT GORDON. I AN TRYING TO FIND OUT IF SOLUTIONS ON A AUTOSTAINER HAVE TO BE FILTERED AFTER EACH SPECIMEN ARE PROCESSED. JAMIE DUKES PATHOLOGY DEPT (706)787-8473 From john.mcginley <@t> colostate.edu Thu Dec 16 12:22:01 2004 From: john.mcginley <@t> colostate.edu (John McGinley) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Colon whole mounts and counting aberrant crypt foci (ACF) Message-ID: <200412161818.iBGIIePB024736@neo.agsci.colostate.edu> Hi, Our lab is just beginning to study colon cancer using a rat model and I was wondering if anyone out there has experience with making colon whole mounts and counting aberrant crypt foci (ACF). It looks like the most accepted method in the literature is cut the colon along the longitudinal axis, prep as a whole mount (mucosal side up), wash in PBS, fix in 10% formalin overnight, rinse, stain with methylene blue and look for ACF at x40. One of the problems I've been having is getting the whole mount to lay flat and adhering to the slide during fixation. I've been blotting the serosal layer with kimwipes to remove excess fluid prior to making the whole mount on plain glass slides. Some reports in the literature indicate that the colon is mounted or pinned to a cork board during fixation to help it lay flat, stain while free floating and then transfer to a slide for microscopic evaluation. This seems like a lot of work and I would rather mount the tissue to a slide once and be done with it. Should I try using silane treated or poly-lysine slides instead? When looking for ACF under the microscope should the whole mount be cover slipped using aqueous mounting media or just left wet? What is the best method for counting the number of ACF? Should I use a reticle, a cm grid transparency overlay on the tissue, etc.? Any information you can provide would be appreciated. Thanks in advance. Regards, John ----------------------------------- John N. McGinley, HTL(ASCP) Cancer Prevention Lab Colorado State University 111 Shepardson bldg. 1173 Campus Delivery Fort Collins, CO 80523-1173 Ph: (970) 491-3041 Fx: (970) 491-3542 Email: john.mcginley@colostate.edu Web: www.cpl.colostate.edu From gcallis <@t> montana.edu Thu Dec 16 13:25:59 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Re: rabbit bone specimens In-Reply-To: <5.1.0.14.2.20041216110632.00a7be60@pop.service.ohio-state. edu> References: <5.1.0.14.2.20041216110632.00a7be60@pop.service.ohio-state.edu> Message-ID: <6.0.0.22.1.20041216111701.01b66cd0@gemini.msu.montana.edu> Some questions first: Are you working with whole femurs and tibias OR do you bisect the samples? Cutting into slabs is also possible using a diamod cutoff blade i.e. Buehler Isomet. This should be done BEFORE fixation to speeds up fixation and eventually speed up decalcification by simple size reduction. There was an excellent article in the old Stain Technology about working with rabbit bones some years back. Ohrt, C. et al. Preparation of whole rabbit knee joints for microscopy examination, Stain Technology, 61(6):353-360.004, 1986. This article is excellent. You refer to Cal Ex decalcifier, but which one? The dilute HCl Cal Ex is 1.34 M HCl when calculated is 8.3% HCl and is rather strong!! Decalcification in this solution will more than likely result in overexposure to acid aka overdecalcification, and will damge the bone, and nuclei in all cell types. Bones must be totally fixed before going into any acid, so fix whole bones for a week or longer, and change fixative after 1st or second day to replenish fixing agent. If the Cal-Ex is the fixative/formic acid combination, check the MSDS as the formic acid may be close to 10%? Two weeks in this solution, if a whole bone, may result in some autolysis, as this decalcifier is designed for tiny bone biopsies. It would be better to fix in formalin then decalcify in a formic acid decalcifier, particularly for that period of time. Do you do decalcification endpoint determinations? If not, 2 weeks in either of these decalcifiers could result in overexposure to acids and damage bone, soft tissues. Formic acid can be just as damaging to tissues if the bones are left in too long. Rabbit bone can be difficult since the articular cartilage is thinner than some species. When you process for longer period of time, the cartilage becomes very dry since water bound to protein in cartilage is removed along with free water, not ideal. The bone however, tends to be ok due to its density but needs the longer processing for just that reason, in order to dehydrate, clear and infiltrate. How to compensate: After facing the block, lay block on an ice block with water on top for a few minutes, NOT a long time or tissue will swell out of block. The section should be very flat, without compression as it comes off the blade edge. Using Tissue Prep 2, a hard paraffin helps support bone with cartilage, or some other harder paraffin. IF your cartilage continues to look wrinkled or did not flatten out on water bath, you can try several things. One, lay section on cold 10% alcohol, pick up and go to warm water bath to let it flatten - be careful the sections are not exploding, so use cooler waterbath i.e 41C. Second: try Tween 20 (not sure of concentration, others have used this) in the waterbath, not a great deal, but just enough to break surface tension on water, and let section flatten then pick up. Third: use a waterbath that is 5% DMSO (not great to put hands in or breathe) but this will flatten cartilage wonderfully. Perfectly flat cartilage on slide surface is crucial for section adherence to slide surface. Some gelatin sub slides (don't do this to pricey plus charge, but presub clean, regular slides, then pick up sections onto these). When you dry slide, do it at 37C to 40C keeping slides FLAT after a blot and good vertical draining. Dry overnight but longer is preferred here - several days. Drying bone w/cartilage sections at 60C continues to dry out cartilage even more, and when you stain, the section releases and curls up. BEWARE of harsh running tap water rinsing!! Gentle water rinsing only, and do not use ammonia water as bluing agent, it may cause section to release. As for processing, we did 1 1/2 hours for thinner slabs (4 -5 mm) and bisected in half knees for 2 hours per station with minimum of 3 paraffin changes always with vacuum and pressure. A popular schedule here is 70%, 80%, 95% x 2, 100% x 2, xylene x 1, Clearite 3 x 1, paraffin X 3 (we do 4 changes). The xylene in first change insures clearing with no residual, and Clearite 3 is less hardening to bone. You can use Propar and you can do two changes of these substitutes, have tried them all successfully with larger bone. Use the best, sharpest blades possible, high profiles are sturdier and just as sharp as low profile, but cut dense bone better without chatter problems. Change to new sharp edge frequently, this is often done between ribbons here and even and blocks. That sharp edge is a must. The bunny can be more difficult, but good sections are possible. Just try some of suggested tweaks. Vyou wrote: >I am working on a research projects with rabbit femurs and tibias. They >are decalcified in cal-ex from fisher for 2 weeks and then processed for >1.5 hours in each solution on the processor. 2 changes of 70, 95, 100%, >xylene and then paraffin. I am really having problems with the cartilage >folding right at the curve of the specimen. It seems as though the bone >just below the cartilage is not thoroughly processed or hard. When I do >feel I get a decent section it seems to fold when I stain. I am using >plus slides. I don't normally do a lot of research so if anyone can help >me with this problem either with cutting or processing solutions I would >really appreciate the help. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Stephanie.Williams <@t> choa.org Mon Dec 13 13:12:01 2004 From: Stephanie.Williams <@t> choa.org (Williams, Stephanie) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Children's Healthcare of Atlanta has an opening for a Histotechnician. Message-ID: <8CC363747C897849B9FBD5B24F98F0C0048C5331@CHOAMAIL1.choa.org> Children's Healthcare of Atlanta has an opening for a Histotechnician. This is a Monday - Friday day shift position at Children's at Egleston. Children's Healthcare of Atlanta is one of the country's leading pediatric healthcare organizations with 2 pediatric hospitals and 16 satellite locations. The Atlanta Business Chronicle has named Children's the #1 Healthcare Employer in Atlanta. We use equipment specifically designed for children and address their psychological needs through family-centered care. You'll have the opportunity to enhance the lives of children through excellence in patient care, research and education while working on interdisciplinary teams in a close-knit, supportive environment. Children's offers a Total Rewards Program that includes: * Competitive compensation * Flexible scheduling * Health insurance including dental and vision plans * Retirement plans with generous vesting and employer contributions * Up to five weeks of paid time off each year including holidays * Childcare programs * Educational assistance * Professional development and more. QUALIFICATIONS: * Certification as a Histotechnician (ASCP or equivalent) required * An Associates Degree or higher in Biology, Chemistry, or life sciences in addition to the certification is preferred * 2 years previous experience in a clinical laboratory is preferred * Eligible for classification as a Histotechnician by Georgia Department of Human Resources * Requires limited use of independent judgement and responsibility with minimum technical supervision * Candidate needs strong background in processing, embedding, cutting, staining, and performing frozen sections * Must also have a strong background in special stains, particularly fungal and microbial stains; experience with processing cytology specimens is desired as well Stephanie Williams Senior Recruiter Children's Healthcare of Atlanta phone 404.785.7818 fax 404.785.7799 www.choa.org From lmclemor <@t> seattlecca.org Thu Dec 16 14:27:33 2004 From: lmclemor <@t> seattlecca.org (McLemore, Lisa M) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Re: Histology Salaries Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B948A909BF@wala01.seattlecca.org> Hello all, Recently, there was someone who had just passed her HT exam inquiring about salary. The salaries HT's make depend on many variables. ( i.e.. where you work, what city your in, experience etc...) However, for FYI there is a web site that you can visit (www. salary.com) to get a general idea of what the market is like. Go to job category and select "Healthcare Technicians" and it will take you to "Job Titles". There you will be able to view graphs and basic reports for your area. Hope this helps. Lisa M. McLemore Lead Histologist Research Histopathology Shared Resources Fred Hutchinson Cancer Research Center Phone: (206) 288-6898 Fax: (206) 288-1345 lmclemor@seattlecca.org This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From jcline <@t> wchsys.org Thu Dec 16 14:31:16 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Claudia Melidona/K903/P63 Message-ID: <000801c4e3ae$31a13680$1d2a14ac@wchsys.org> I use the P504S+P63 made by Biocare Medical. I have a Ventana and their predilute works beautifully. ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From NBerghoff <@t> cvm.tamu.edu Thu Dec 16 17:17:11 2004 From: NBerghoff <@t> cvm.tamu.edu (Nora Berghoff) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Russell-Movat's stain - again Message-ID: Hello everyone, is there really nobody who is familiar with the Russell's modification of Movat's pentachrome and who may have an idea as to what the problem may be....? Here is the problem (previous e-mail) again, just in case (I am not giving up yet): **** I have a question about Russell's modification of Movat's pentachrome stain. (Ref.: Russell, HK, "A Modification of Movat's Pentachrome stain", Arch Path, Vol. 94, Aug 1972, 187-191) I have been staining canine intestinal sections that have been fixed in 10% neutral buffered formalin, embedded in paraffin and cut at 5um. The stain worked pretty well and gave good results for a while. Last week I have had the following problem: After destaining the crocein scarlet-acid fuchsin with 5% phosphotungstic acid and then rinsing it in 0.5% acetic acid, it kept destaining the tissue until everything was just very pale red (one slide was completely destained - no red stain left whatsoever and the other stains were pale as well). I always control the destaining under the microscope and stop when there is still intense red color. The color kept coming off the slide in the acetic acid solution and also in the following rinses in absolute alcohol. I make the solutions (phosphotungstic acid and acetic acid) fresh right before I need them. Does anyone have an idea what the problem might be? Is the acetic acid not strong enough/the phosphotungstic acid too strong, etc?? Is it a pH problem? **** As I previously said, I have tried to find an answer somewhere else before bothering this e-mail list, but I simply can't find anything. And I am afraid I am not experienced enough to detect simple errors that may be obvious to someone else... So if there is anyone who has even a vague idea of what may cause these problems, PLEASE reply...My samples are very valuable, which is why I don't want to keep staining (and then discarding) sections unnecessarily, just hoping for the problem to resolve on its own. Thank you!!! Nora Nora Berghoff Research Assistant Texas A&M University College Station, TX From AnthonyH <@t> chw.edu.au Thu Dec 16 17:43:38 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Russell-Movat's stain - again Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E147@simba.kids> Nora, I would wonder at the strength of the acids you are using. It could be they are stronger than you are used to using. Also possibly the ethanols used for dehydrating may be contaminated with acid. Have you changed dyes? Check the CI and the % dye purity. Just some thoughts Regards Tony Henwood JP, 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: Nora Berghoff [mailto:NBerghoff@cvm.tamu.edu] Sent: Friday, 17 December 2004 10:17 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Russell-Movat's stain - again Hello everyone, is there really nobody who is familiar with the Russell's modification of Movat's pentachrome and who may have an idea as to what the problem may be....? Here is the problem (previous e-mail) again, just in case (I am not giving up yet): **** I have a question about Russell's modification of Movat's pentachrome stain. (Ref.: Russell, HK, "A Modification of Movat's Pentachrome stain", Arch Path, Vol. 94, Aug 1972, 187-191) I have been staining canine intestinal sections that have been fixed in 10% neutral buffered formalin, embedded in paraffin and cut at 5um. The stain worked pretty well and gave good results for a while. Last week I have had the following problem: After destaining the crocein scarlet-acid fuchsin with 5% phosphotungstic acid and then rinsing it in 0.5% acetic acid, it kept destaining the tissue until everything was just very pale red (one slide was completely destained - no red stain left whatsoever and the other stains were pale as well). I always control the destaining under the microscope and stop when there is still intense red color. The color kept coming off the slide in the acetic acid solution and also in the following rinses in absolute alcohol. I make the solutions (phosphotungstic acid and acetic acid) fresh right before I need them. Does anyone have an idea what the problem might be? Is the acetic acid not strong enough/the phosphotungstic acid too strong, etc?? Is it a pH problem? **** As I previously said, I have tried to find an answer somewhere else before bothering this e-mail list, but I simply can't find anything. And I am afraid I am not experienced enough to detect simple errors that may be obvious to someone else... So if there is anyone who has even a vague idea of what may cause these problems, PLEASE reply...My samples are very valuable, which is why I don't want to keep staining (and then discarding) sections unnecessarily, just hoping for the problem to resolve on its own. Thank you!!! Nora Nora Berghoff Research Assistant Texas A&M University College Station, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From bdyparts <@t> zoominternet.net Thu Dec 16 19:28:43 2004 From: bdyparts <@t> zoominternet.net (Tony Green) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Re: Ventana Benchmark References: <200412150858.iBF8whCJ032404@mx-1.zoominternet.net> Message-ID: <004701c4e3d7$beed19e0$f861ef18@S0027610805> Our lab also considered acquiring a Ventana Benchmark (new model) so that we could also do routine IHC's and HPV testing on thin preps and found that it wasn't to difficult. However, I was warned that with the new unit the ER/PR was a little tricky and if you are going to do HER2/neu also check the FDA regulations. There was something about DAKO being the only current FDA approved company if your including the results on a patient's report to be used as a prognostic indicator. Please understand, this is what I was told when I asked around just like you are doing. I didn't research it because we found that in our situation, it was better that we maintained with DAKO for financial and personnel reasons. I do believe that DAKO has a detection system for HPV ISH. I'm not sure what it all entails though. Hope this helps. Tony From jkiernan <@t> uwo.ca Thu Dec 16 23:55:14 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Russell-Movat's stain - again (LONG) References: Message-ID: <41C274C2.591A8E9A@uwo.ca> Dear Nora & all, I've never done the method, but have looked it up in Bancroft & Gamble, and also looked at the properties of the dyes, so here are some facts and suggestions. The name crocein scarlet is applied to at least 4 similar anionic disazo dyes (CI 26905, CI 27155, CI 27160 and CI 27165). They all have various synonyms. Acid fuchsine is an anionic triphenylmethane dye with molecules about the same size (MW 550-600 or so), so all these dyes should have similar staining properties. The action of phosphotungstic acid (PTA) is to attach to collagen, expelling the red anionic dye(s) from the collagen fibres but not from cytoplasm, fibrin etc. The exact mechanism by which PTA does this is debatable. The sections are rinsed in weak acetic acid rather than water to prevent extraction of the anionic red dyes from stained structures. Water counts as slightly alkaline where dye anions are concerned. YOUR PROBLEM: In being extracted by acetic acid, your red dyes are behaving like cationic rather than anionic dyes. One possible explanation might be that the triphenylmethane dye is basic fuchsine rather than acid fuchsine. As a cationic dye, basic fuchsine is extracted from tissues by acids. If your crocein scarlet + acid fuchsine solution contains an excess of basic fuchsine cations over crocein scarlet anions, the net result will probably be that the sections are being stained only with basic fuchsine. Unfortunately there's a typo in Bancroft & Gamble (p.160) and the prescribed amount of acid fuchsine for the mixture is omitted, so I can't work out the molar amounts of crocein scarlet and acid (or basic) fuchsine. All this is speculation, of course, based on the simple rule that simple acids extract basic dyes, not acid dyes from stained tissues. (There may be some exceptions, expecially among dye-metal complexes in stains such as haemalums and iron-haematoxylins. Also, PTA is not a simple acid like acetic or HCl; it has complicated reactions and interactions with dyes of different types.) Making up a solution with basic fuchsine instead of acid fuchsine is an easily made mistake because there's quite a range of names to be found on bottle labels: Fuchsin, acid; Fuchsin, special for DNA; Fuchsin basic, special for flagella etc etc. They nearly always omit the terminal e of fuchsine too. That e is there for a good reason. You won't find "fuchsin" in American (Webster's) or English (Oxford; Chambers') dictionaries. A more serious possibility is that one or both of the red dyes is not what the label on the bottle says it is. Does the acid fuchsin(e) bottle carry a Biological Stain Commission (BSC) certification sticker? It's a little square label that includes a reproduction of the late H.J.Conn's signature. Every batch of a dye certified by the BSC has passed several chemical tests and has also been found satisfactory in staining methods that require correctly identified dyes with adequate dye-content and a low level of coloured impurities. Nearly all dye powders contain non-dye ingredients such as dextran, NaCl, Na2SO4 etc, and also coloured impurities that are by-products of the complex chemical reactions that generate synthetic dyes. Some dyes sold as stains are not what they claim to be, and they do not work. This was a major problem, especially in the USA, in the first half of the 20th century, and it is one of the reasons for the existence of the Biological Stain Commission. The BSC has been testing stains since 1925. In the early 1990s the BSC detected fake dyes purporting to be light green SF (used in the Papanicolau method for cervical cytology smears). The matter was soon resolved, and reported in the BSC's journal, Biotechnic & Histochemistry, which is not expensive, even for an individual subscriber. I hope this advice helps. If dye identity is a problem, seek advice from the BSC. http://www.biostains.org _________________________________ John Kiernan London, Canada. _____________________________________- Nora Berghoff wrote: > > Hello everyone, > > is there really nobody who is familiar with the Russell's modification > of Movat's pentachrome and who may have an idea as to what the problem > may be....? > > Here is the problem (previous e-mail) again, just in case (I am not > giving up yet): > > **** > I have a question about Russell's modification of Movat's pentachrome > stain. > (Ref.: Russell, HK, "A Modification of Movat's Pentachrome stain", Arch > Path, Vol. 94, Aug 1972, 187-191) > > I have been staining canine intestinal sections that have been fixed in > 10% neutral buffered formalin, embedded in paraffin and cut at 5um. > > The stain worked pretty well and gave good results for a while. Last > week I have had the following problem: > > After destaining the crocein scarlet-acid fuchsin with 5% > phosphotungstic acid and then rinsing it in 0.5% acetic acid, it kept > destaining the tissue until everything was just very pale red (one slide > was completely destained - no red stain left whatsoever and the other > stains were pale as well). > I always control the destaining under the microscope and stop when > there is still intense red color. > The color kept coming off the slide in the acetic acid solution and > also in the following rinses in absolute alcohol. > I make the solutions (phosphotungstic acid and acetic acid) fresh right > before I need them. > > Does anyone have an idea what the problem might be? Is the acetic acid > not strong enough/the phosphotungstic acid too strong, etc?? Is it a pH > problem? > **** > > As I previously said, I have tried to find an answer somewhere else > before bothering this e-mail list, but I simply can't find anything. And > I am afraid I am not experienced enough to detect simple errors that may > be obvious to someone else... > So if there is anyone who has even a vague idea of what may cause these > problems, PLEASE reply...My samples are very valuable, which is why I > don't want to keep staining (and then discarding) sections > unnecessarily, just hoping for the problem to resolve on its own. > > Thank you!!! > Nora > > Nora Berghoff > Research Assistant > Texas A&M University > College Station, TX > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Chewy71874 <@t> aol.com Fri Dec 17 00:38:58 2004 From: Chewy71874 <@t> aol.com (Chewy71874@aol.com) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Kappa and Lambda Message-ID: We are having a problem with background staining on Kappa and Lambda stains. We use DAKO rabbit polyclonal antibodies, DAKO TRS for antigen retrieval in the steamer, Envision+ HRP, DAB. The background staining is especially bad on Bouin's fixed, decaled bone marrows. Does anyone else have this problem? Do you get cleaner staining with monoclonal antibodies? We are open to suggestions. Thanks in advance, Ellen Yee, HT Lead Tech, Immuno Dept. Central Histology Facility Diagnostic Pathology Medical Group 2420 J St. Sacramento, CA 95816 ph: 916-447-2718 fax: 916-447-0620 Sacramento, CA From psanquin <@t> lugo.usc.es Fri Dec 17 04:45:09 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Paraffin sections artifact Message-ID: <3.0.6.32.20041217114509.007ec450@pop.lugo.usc.es> Dear Histonetters, Once more time I need your help. I have a disgusting problem with my mouse's brain paraffin-embedded sections. Right after the sectioning they seem ok but after several days stored at 37?C they develope a dreadful artifact. Plenty of holes appear randomly placed in the whole section. The appearance is like a Gruyere Cheese. Have you seen an artifact of this kind? The cause should probably either a bad fixation or a bad embedding. Asuming the first I ask to myself: If the water of the tissue has been removed in the embedding how can take place any spoiling process in the tissue? Thanks in advance for your help Pablo Sanchez From balajimr <@t> drreddys.com Fri Dec 17 04:55:28 2004 From: balajimr <@t> drreddys.com (balajimr@drreddys.com) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Testes - Fixation, Staining, Staging Message-ID: Dear Histonetters, We are in process of standerdising staging of spermatogenesis in Rats.I have following questions Which is the best fixative?Bouin's or modified Davidson's (MD)? Recently an STP recommendation says MD is the best suited. What is the best fixation duration in Bouins fluid?Do we need to wash the same in 70% alcohol after fixation in Bouins? If so please let me know for how long? Does anybody know the composition of MD? I would like to know the procedure of PAS -H & E staining for testes for acrosome demonstration. Can somebody tell me a good reference for staging of the spermatogenesis in Rats? Also aI would like to know a good CD rom available for the same. Dr. M.R. Balaji Dept. Pre clinical safety evaluation, Discovery research, Dr. Reddys Laboratories ltd. Bollaram Road, Miyapur, Hyderabad, 500 049 Andhra Pradseh, INDIA Email - balajimr@drreddys.com Tel: 040- 23045439 - Ext.432. From balajimr <@t> drreddys.com Fri Dec 17 04:55:28 2004 From: balajimr <@t> drreddys.com (balajimr@drreddys.com) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Testes - Fixation, Staining, Staging Message-ID: Dear Histonetters, We are in process of standerdising staging of spermatogenesis in Rats.I have following questions Which is the best fixative?Bouin's or modified Davidson's (MD)? Recently an STP recommendation says MD is the best suited. What is the best fixation duration in Bouins fluid?Do we need to wash the same in 70% alcohol after fixation in Bouins? If so please let me know for how long? Does anybody know the composition of MD? I would like to know the procedure of PAS -H & E staining for testes for acrosome demonstration. Can somebody tell me a good reference for staging of the spermatogenesis in Rats? Also aI would like to know a good CD rom available for the same. Dr. M.R. Balaji Dept. Pre clinical safety evaluation, Discovery research, Dr. Reddys Laboratories ltd. Bollaram Road, Miyapur, Hyderabad, 500 049 Andhra Pradseh, INDIA Email - balajimr@drreddys.com Tel: 040- 23045439 - Ext.432. From Kemlo.Rogerson <@t> elht.nhs.uk Fri Dec 17 05:06:46 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Paraffin sections artifact[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EDA4@bhrv-nt-11.bhrv.nwest.nhs.uk> I would think it's a bit of both. Basically the fluids and wax have not penetrated the brain tissue and the holes are where the tissue has decomposed. Probably! -----Original Message----- From: Pablo S?nchez Quinteiro [mailto:psanquin@lugo.usc.es] Sent: 17 December 2004 10:45 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Paraffin sections artifact[Scanned] Dear Histonetters, Once more time I need your help. I have a disgusting problem with my mouse's brain paraffin-embedded sections. Right after the sectioning they seem ok but after several days stored at 37?C they develope a dreadful artifact. Plenty of holes appear randomly placed in the whole section. The appearance is like a Gruyere Cheese. Have you seen an artifact of this kind? The cause should probably either a bad fixation or a bad embedding. Asuming the first I ask to myself: If the water of the tissue has been removed in the embedding how can take place any spoiling process in the tissue? Thanks in advance for your help Pablo Sanchez _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jl <@t> eyepath.ku.dk Fri Dec 17 05:26:51 2004 From: jl <@t> eyepath.ku.dk (Jens Lindegaard) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Porcine anti-MMP2 Message-ID: <37AD14B3F0F@eyepath.ku.dk> Hello Does anybody have any experience with the use of antibodys against MMP-2 in pigs. What antibody do you use, and where can I get them? Best regards Jens Lindegaard M.D. Eye Pathology Institute Frederik V's Vej 11, 1.floor DK-2100 Copenhagen Denmark Phone +45 3532 6083 Fax +45 3532 6080 From bettina.hutz <@t> orionpharma.com Fri Dec 17 06:18:17 2004 From: bettina.hutz <@t> orionpharma.com (bettina.hutz@orionpharma.com) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Testes - Fixation, Staining, Staging Message-ID: <243E1A79AF5D89438C70513DBADAA4F0019E4034@sfies-exchange1.orionnet.org> Hello, Here instructions for HE and PAS after Hotchkiss my Pathologists likes;) We fix testis in Boiun for 24 hours, then transfer them to 70% ethanol in which they can be kept as long as you wish. We do not rinse them several times in 70% ethanol. If you wish I can ask him more about satging when he returns from Christmas vacation. Hope this helpes you:) I can send you also pictures of my stainings if you like to see the result. Bettina Hutz Research Assistant Department of Discovery Biology Orion Corporation, ORION PHARMA Orionintie 1, P.O.Box 65, FIN-02101 Espoo, Finland Tel. +358 10 429 2607, Fax +358 10 429 2924 Email: bettina.hutz@orionpharma.com Department of Discovery Biology Hemalaun-Eosin staining of paraffin slides 1. Staining procedure Deparaffination 3x xylene each min. 3 minutes 2x ethanol 100% short ethanol 90% short ethanol 80% short ethanol 70% short aqua dest. Short Staining Mayer?s Hemalaum 3 minutes Running tap water 10 minutes Rinse with aqua-dest. Eosin 40 seconds Dehydration & mounting ethanol 70% short ethanol 90% short 2x ethanol 100% short 3x xylene short Mount with xylene based medium 2. Solutions 2.1. Mayer?s Hemalaunn 2.1.1 Stock solution Mayer?s Hemalaum (Fa. Merck) 2.1.2. Usage solution Mayer?s Hemalaun 3 parts Aqua-dest. 1 part Filtrate before use 2.2 Eosin 2.2.1 Stock solution Eosin Yellow 1,0 g Aqua-dest. 1000 ml 2.2.2. Usage solution Eosin stock solution 50,0 ml Aqua dest. 450 ml + 6 drops actetic acid 100% 3. Results Nuclei - blue background - red PAS reaction after Hotchkiss for paraffin slides 1. Staining procedure Deparaffination 3x xylene each min. 3 minutes 2x ethanol 100% short ethanol 90% short ethanol 80% short ethanol 70% short Staining Periodic acid (oxidation) 5 minutes 70% ethanol 1 minute Reduction solution 1 minute 70% ethanol 1 minute Schiff?s reagent 20 minutes Running tap water 10 minutes Mayer?s hemalaun 3 minutes Running tap water 10 minutes Dehydration & mounting ethanol 70% short ethanol 90% short 2x ethanol 100% short 3x xylene short Mount with xylene based medium 2. Solutions 2.1. Mayer?s Hemalaunn 2.1.1 Stock solution Mayer?s Hemalaum (Fa. Merck) 2.1.2. Usage solution Mayer?s Hemalaun 3 parts Aqua-dest. 1 part Filtrate before use 2.2 Periodic acid (produce short before use) Periodic Acid 1,6 g Aqua-dest. 40 ml Ethanol 100% p.a. 140 ml Sodiumacetate 0,2 N 20 ml 2.3 Reduction solution Potassiumiodid 4 g Sodiumthiosulfate 4 g Aqua-dest. 80 ml HCl, 2 N 2 ml 2.4 Schiff reagent Schiff?s reagent (Fa. Sigma-Aldrich), 500 ml 3. Results Nuclei - blue Carbohydrates - pink-deep red From bettina.hutz <@t> orionpharma.com Fri Dec 17 06:18:17 2004 From: bettina.hutz <@t> orionpharma.com (bettina.hutz@orionpharma.com) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Testes - Fixation, Staining, Staging Message-ID: <243E1A79AF5D89438C70513DBADAA4F0019E4034@sfies-exchange1.orionnet.org> Hello, Here instructions for HE and PAS after Hotchkiss my Pathologists likes;) We fix testis in Boiun for 24 hours, then transfer them to 70% ethanol in which they can be kept as long as you wish. We do not rinse them several times in 70% ethanol. If you wish I can ask him more about satging when he returns from Christmas vacation. Hope this helpes you:) I can send you also pictures of my stainings if you like to see the result. Bettina Hutz Research Assistant Department of Discovery Biology Orion Corporation, ORION PHARMA Orionintie 1, P.O.Box 65, FIN-02101 Espoo, Finland Tel. +358 10 429 2607, Fax +358 10 429 2924 Email: bettina.hutz@orionpharma.com Department of Discovery Biology Hemalaun-Eosin staining of paraffin slides 1. Staining procedure Deparaffination 3x xylene each min. 3 minutes 2x ethanol 100% short ethanol 90% short ethanol 80% short ethanol 70% short aqua dest. Short Staining Mayer?s Hemalaum 3 minutes Running tap water 10 minutes Rinse with aqua-dest. Eosin 40 seconds Dehydration & mounting ethanol 70% short ethanol 90% short 2x ethanol 100% short 3x xylene short Mount with xylene based medium 2. Solutions 2.1. Mayer?s Hemalaunn 2.1.1 Stock solution Mayer?s Hemalaum (Fa. Merck) 2.1.2. Usage solution Mayer?s Hemalaun 3 parts Aqua-dest. 1 part Filtrate before use 2.2 Eosin 2.2.1 Stock solution Eosin Yellow 1,0 g Aqua-dest. 1000 ml 2.2.2. Usage solution Eosin stock solution 50,0 ml Aqua dest. 450 ml + 6 drops actetic acid 100% 3. Results Nuclei - blue background - red PAS reaction after Hotchkiss for paraffin slides 1. Staining procedure Deparaffination 3x xylene each min. 3 minutes 2x ethanol 100% short ethanol 90% short ethanol 80% short ethanol 70% short Staining Periodic acid (oxidation) 5 minutes 70% ethanol 1 minute Reduction solution 1 minute 70% ethanol 1 minute Schiff?s reagent 20 minutes Running tap water 10 minutes Mayer?s hemalaun 3 minutes Running tap water 10 minutes Dehydration & mounting ethanol 70% short ethanol 90% short 2x ethanol 100% short 3x xylene short Mount with xylene based medium 2. Solutions 2.1. Mayer?s Hemalaunn 2.1.1 Stock solution Mayer?s Hemalaum (Fa. Merck) 2.1.2. Usage solution Mayer?s Hemalaun 3 parts Aqua-dest. 1 part Filtrate before use 2.2 Periodic acid (produce short before use) Periodic Acid 1,6 g Aqua-dest. 40 ml Ethanol 100% p.a. 140 ml Sodiumacetate 0,2 N 20 ml 2.3 Reduction solution Potassiumiodid 4 g Sodiumthiosulfate 4 g Aqua-dest. 80 ml HCl, 2 N 2 ml 2.4 Schiff reagent Schiff?s reagent (Fa. Sigma-Aldrich), 500 ml 3. Results Nuclei - blue Carbohydrates - pink-deep red From eileen_dusek <@t> yahoo.com Fri Dec 17 08:27:23 2004 From: eileen_dusek <@t> yahoo.com (eileen dusek) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] coverslipping film Message-ID: <20041217142723.60336.qmail@web11908.mail.yahoo.com> Good Morning, Does anyone know how many slides a roll of coverslipping film will cover. I have to do a cost analysis for the lab. Thanks Happy Holiday Eileen Edward Hospital --------------------------------- Do you Yahoo!? All your favorites on one personal page – Try My Yahoo! From djohnson14 <@t> hotmail.com Fri Dec 17 09:09:43 2004 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] coverslipping film In-Reply-To: <20041217142723.60336.qmail@web11908.mail.yahoo.com> Message-ID: Eileen I have been told that the KP Tape from Mercedes Medical which is 70 meters long will cover about 1500 slides. The other tape on the market is 60 meters so i am not sure how many that covers Thanks Dave Johnson Mercedes Medical 800-331-2716 x157 >From: eileen dusek >To: histonet >Subject: [Histonet] coverslipping film >Date: Fri, 17 Dec 2004 06:27:23 -0800 (PST) > >Good Morning, >Does anyone know how many slides a roll of coverslipping film will cover. I >have to do a cost analysis for the lab. >Thanks > >Happy Holiday >Eileen >Edward Hospital > > >--------------------------------- >Do you Yahoo!? > All your favorites on one personal page – Try My Yahoo! >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emerald_lake77 <@t> yahoo.com Fri Dec 17 09:21:38 2004 From: emerald_lake77 <@t> yahoo.com (- -) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Need assistance with in situ protocol!!! (LONG) Message-ID: <20041217152138.20291.qmail@web42302.mail.yahoo.com> Dear histonetters, I am in dire need of having my in situ protocol work!! I cannot find the problem and feel there may be too many to fix the situation on my own. Please if you are up for the challenge, look over my protocol and problems I am facing and offer up any suggestions or changes I should make. Also, does anyone know of a good DIG-labeled RNA probe that a particular manufacturer makes and that can be used for targets found throughout different mouse tissues -- just trying also to ensure protocol is wrong and not probe?? Thank you. Note: Letter ‘u’ used to represent micro. Temps given in celcius. Sorry – can’t tell you what probe I was using. Only that it is a DIG-labeled RNA probe. Problems: Antisense and sense appear to both stain. Large amount of background (endogenous peroxidase?) At the end of my washes (after hybridization) tissue begins to disintegrate and cells begin falling away from the sections in large clumps – only tissue I am using is mouse kidney. The kidney is perfused (saline then 4% PF/PBS – RNase free) and allowed to sit overnight in 4% PF/PBS. Afer switching the 70% alcohol for ~3-5 hours, the tissues are routinely processed and embedded. Sections are sectioned – 5 um, and placed onto positively charged barrier slides (Biogenex). They incubate on a plate (covered) at 37 degrees and then before deparaffinizing are baked at 55 degrees for 1 hour. At all times RNase free glassware/solutions are used during the entire process. Bench tops and surrounding area wiped with RNase Away. PROTOCOL: Slides deparaffinized and brought down to distilled water, then washed 2x (5mins each) in 1x PBS. Proteinase K digestion: Slides incubated in 0.1M Tris and 50mM EDTA (pH 8) prewarmed at 37 degrees containing 5 ug/ml Prot. K. Incubation time 17mins (determined to be optimal). After incubation, slides immersed in 0.1M glycine/PBS for 5 mins. The slides were then post-fixed in 4% paraformaldehyde/PBS for 3 mins. Rm Temp. Rinse with 1x PBS 2x (5 mins) Acetylation Slides placed in a glass staining jar containing 0.1M triethanolamine and while stirring, acetic anhydride was added to a final concentration of 0.25% acetic anhydride. Slides rinse in distilled water and then dried on hot plate (37-40 degrees) for ~10-15 mins. Hybridization Buffer: Frozen at –20 degrees. Placed in 50 degree waterbath before use. Reagents Final Concentration Deionized formamide 50% 20x SSC 5x Dextran sulfate 10% 100x denhardt’s solution 5x 10% SDS 2% 10mg/ml denatured 100ug/ml sheared salmon DNA Sterile ddH2O Aliquots of 900ul made up and frozen. Dilute riboprobe in buffer to final concentration of 2.5ng/ul ( DIG-label RNA probe) To each section between 40-60 ul of probe was added to cover section – more for larger sections. Sections placed in hybridization chamber (Boekel) at 42 degrees, covered each section with Parafilm to avoid evaporation. Slides incubated at above temp. 20 hours overnight. Chamber kept humid with 5x SSC in the provided wells. Checked slides next day to ensure moisture still under parafilm and that there was no evaporation – none seen. Slides placed in 2x SSC/0.1%SDS for 2mins. Parafilm begins to float off to liquid surface. Slides washed in 2x SSC/0.1% SDS at room temp shaking gently 2x (5 mins each) Wash slides 0.1x SSC/0.1%SDS at the same temperature used for hybridization (prewarmed at 42 degrees) -- 2x 10mins each Rinsed briefly in 2x SSC twice (Room Temp) to remove traces of SDS Slides incubated with 10ug/ml RNase solution in 2x SSC at 37 degrees for 15 mins to selectively degrade single stranded RNA and reduce non-specific signal. Wash in 2x SSC/ 50%formamide for ½ hr. at 52 degrees gently shaking Rinse briefly in 2x SSC Rm. temp. Placed into Buffer 1: 80ml 1 M Tris-HCl pH 7.5 40ml 3M NaCl Add autoclaved deionized water to 800ml Rinsed briefly Place into fresh Buffer 1 for 5 mins. Place into Buffer 2 for 30 mins. shaking Rm. Temp. Buffer 2 1 M Tris HCl pH 7.5 3M NaCl Triton X-100 (0.5%) 10% Blocking Reagent (1% final concentration) Blocking reagent bought from Roche (final concentration is 1% or 1x) Blocking Reagent Cat. No. 1 096 176 – made up as instructed by manufacturer. Sheep Anti-DIG-AP antibody used (Roche) –0- Cat. No. 1 093 274 Buffer 2 tapped off and 100-150ul of antibody, diluted 1:50 in 6ml 2% normal sheep serum/0.5% Triton X-100 in PBS was added. Slides incubated with antibody at 37 degrees for 2 hours. Slides washed 3x in PBS (5 mins. each) Rinsed in 100mM Tris-HCl (pH 9.5) In darkness make up BCIP/NBT solution while rinsing in above Tris. BCIP/NBT provided by DAKO – new kit, made up as specified. Difficult to make mistake here. Added BCIP/NBT solution 3-4 drops to cover section. After 20-30 mins. Slides checked for signal. Little seen except for tissue coming off slides (clumps of cells) and floating off. Allowed to sit overnight at 4 degrees – next day there was signal – very strong in some slides – however, there was background, and sense and antisense slides had what appeared to be signal. ??? Any suggestions??? THANK YOU FOR YOUR ASSISTANCE --------------------------------- Do you Yahoo!? Yahoo! Mail - Easier than ever with enhanced search. Learn more. From Kemlo.Rogerson <@t> elht.nhs.uk Fri Dec 17 09:42:35 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] coverslipping film[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EDA5@bhrv-nt-11.bhrv.nwest.nhs.uk> Six sevenths of 1500, which is um........ Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Dave Johnson [mailto:djohnson14@hotmail.com] Sent: 17 December 2004 15:10 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] coverslipping film[Scanned] Eileen I have been told that the KP Tape from Mercedes Medical which is 70 meters long will cover about 1500 slides. The other tape on the market is 60 meters so i am not sure how many that covers Thanks Dave Johnson Mercedes Medical 800-331-2716 x157 >From: eileen dusek >To: histonet >Subject: [Histonet] coverslipping film >Date: Fri, 17 Dec 2004 06:27:23 -0800 (PST) > >Good Morning, >Does anyone know how many slides a roll of coverslipping film will cover. I >have to do a cost analysis for the lab. >Thanks > >Happy Holiday >Eileen >Edward Hospital > > >--------------------------------- >Do you Yahoo!? > All your favorites on one personal page - Try My Yahoo! >_______________________________________________ >Histonet mailing list >Histonet@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 SJones <@t> cvm.tamu.edu Fri Dec 17 09:56:17 2004 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] salaries for stent labs Message-ID: Hi Netters, I was wondering if anyone would know what the salaries are for technicians that are doing stent work in plastics? Thanks, Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 From Abizar.A.Lakdawalla <@t> appliedbiosystems.com Fri Dec 17 10:06:47 2004 From: Abizar.A.Lakdawalla <@t> appliedbiosystems.com (Abizar A Lakdawalla) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Need assistance with in situ protocol!!! (LONG) In-Reply-To: <20041217152138.20291.qmail@web42302.mail.yahoo.com> Message-ID: Would recommend running the following controls: 1. Slide with no sense probe (but all the other reagents) 2. Slide with no probe, no antibody If Slide 1 and 2 shows staining then you have endogenous AP activity, this is extremely unlikely with all the pretreatment that you do (prot K, acetic anhydride, etc). If only Slide 1 shows staining then the antibody is binding non-specifically to the tissue (again unlikely, but it is better to rule out the possibility). If you do see staining here then the antibody might need to be pre-adsorbed with mouse kidney tissue materials. If both the above slides are negative then the next step would be to titrate the probe concentration and the hybridization and post-hybridization conditions. Simplest would be to run a set of slides with 1/3rd and 1/10th the amount of sense and anti-sense probes and see if the signal intensity decreases with probe concentration. In addition you might need to increase stringency (higher wash temperature) to maximize the difference between sense and anti-sense probes. If none of the above work, then back to the drawing board and re-design the probes (were they checked by BLASTing the Sequences?). Abizar - - Sent by: histonet-bounces@lists.utsouthwestern.edu 12/17/2004 07:21 AM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] Need assistance with in situ protocol!!! (LONG) Dear histonetters, I am in dire need of having my in situ protocol work!! I cannot find the problem and feel there may be too many to fix the situation on my own. Please if you are up for the challenge, look over my protocol and problems I am facing and offer up any suggestions or changes I should make. Also, does anyone know of a good DIG-labeled RNA probe that a particular manufacturer makes and that can be used for targets found throughout different mouse tissues -- just trying also to ensure protocol is wrong and not probe?? Thank you. Note: Letter ?u? used to represent micro. Temps given in celcius. Sorry ? can?t tell you what probe I was using. Only that it is a DIG-labeled RNA probe. Problems: Antisense and sense appear to both stain. Large amount of background (endogenous peroxidase?) At the end of my washes (after hybridization) tissue begins to disintegrate and cells begin falling away from the sections in large clumps ? only tissue I am using is mouse kidney. The kidney is perfused (saline then 4% PF/PBS ? RNase free) and allowed to sit overnight in 4% PF/PBS. Afer switching the 70% alcohol for ~3-5 hours, the tissues are routinely processed and embedded. Sections are sectioned ? 5 um, and placed onto positively charged barrier slides (Biogenex). They incubate on a plate (covered) at 37 degrees and then before deparaffinizing are baked at 55 degrees for 1 hour. At all times RNase free glassware/solutions are used during the entire process. Bench tops and surrounding area wiped with RNase Away. PROTOCOL: Slides deparaffinized and brought down to distilled water, then washed 2x (5mins each) in 1x PBS. Proteinase K digestion: Slides incubated in 0.1M Tris and 50mM EDTA (pH 8) prewarmed at 37 degrees containing 5 ug/ml Prot. K. Incubation time 17mins (determined to be optimal). After incubation, slides immersed in 0.1M glycine/PBS for 5 mins. The slides were then post-fixed in 4% paraformaldehyde/PBS for 3 mins. Rm Temp. Rinse with 1x PBS 2x (5 mins) Acetylation Slides placed in a glass staining jar containing 0.1M triethanolamine and while stirring, acetic anhydride was added to a final concentration of 0.25% acetic anhydride. Slides rinse in distilled water and then dried on hot plate (37-40 degrees) for ~10-15 mins. Hybridization Buffer: Frozen at ?20 degrees. Placed in 50 degree waterbath before use. Reagents Final Concentration Deionized formamide 50% 20x SSC 5x Dextran sulfate 10% 100x denhardt?s solution 5x 10% SDS 2% 10mg/ml denatured 100ug/ml sheared salmon DNA Sterile ddH2O Aliquots of 900ul made up and frozen. Dilute riboprobe in buffer to final concentration of 2.5ng/ul ( DIG-label RNA probe) To each section between 40-60 ul of probe was added to cover section ? more for larger sections. Sections placed in hybridization chamber (Boekel) at 42 degrees, covered each section with Parafilm to avoid evaporation. Slides incubated at above temp. 20 hours overnight. Chamber kept humid with 5x SSC in the provided wells. Checked slides next day to ensure moisture still under parafilm and that there was no evaporation ? none seen. Slides placed in 2x SSC/0.1%SDS for 2mins. Parafilm begins to float off to liquid surface. Slides washed in 2x SSC/0.1% SDS at room temp shaking gently 2x (5 mins each) Wash slides 0.1x SSC/0.1%SDS at the same temperature used for hybridization (prewarmed at 42 degrees) -- 2x 10mins each Rinsed briefly in 2x SSC twice (Room Temp) to remove traces of SDS Slides incubated with 10ug/ml RNase solution in 2x SSC at 37 degrees for 15 mins to selectively degrade single stranded RNA and reduce non-specific signal. Wash in 2x SSC/ 50%formamide for ? hr. at 52 degrees gently shaking Rinse briefly in 2x SSC Rm. temp. Placed into Buffer 1: 80ml 1 M Tris-HCl pH 7.5 40ml 3M NaCl Add autoclaved deionized water to 800ml Rinsed briefly Place into fresh Buffer 1 for 5 mins. Place into Buffer 2 for 30 mins. shaking Rm. Temp. Buffer 2 1 M Tris HCl pH 7.5 3M NaCl Triton X-100 (0.5%) 10% Blocking Reagent (1% final concentration) Blocking reagent bought from Roche (final concentration is 1% or 1x) Blocking Reagent Cat. No. 1 096 176 ? made up as instructed by manufacturer. Sheep Anti-DIG-AP antibody used (Roche) ?0- Cat. No. 1 093 274 Buffer 2 tapped off and 100-150ul of antibody, diluted 1:50 in 6ml 2% normal sheep serum/0.5% Triton X-100 in PBS was added. Slides incubated with antibody at 37 degrees for 2 hours. Slides washed 3x in PBS (5 mins. each) Rinsed in 100mM Tris-HCl (pH 9.5) In darkness make up BCIP/NBT solution while rinsing in above Tris. BCIP/NBT provided by DAKO ? new kit, made up as specified. Difficult to make mistake here. Added BCIP/NBT solution 3-4 drops to cover section. After 20-30 mins. Slides checked for signal. Little seen except for tissue coming off slides (clumps of cells) and floating off. Allowed to sit overnight at 4 degrees ? next day there was signal ? very strong in some slides ? however, there was background, and sense and antisense slides had what appeared to be signal. ??? Any suggestions??? THANK YOU FOR YOUR ASSISTANCE --------------------------------- Do you Yahoo!? Yahoo! Mail - Easier than ever with enhanced search. Learn more. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CMCCOLLOUGH <@t> dnr.state.md.us Fri Dec 17 10:14:41 2004 From: CMCCOLLOUGH <@t> dnr.state.md.us (McCollough, Carol) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] coverslipping film Message-ID: Divide the length of your roll of coverslipping film by the length of the piece that the coverslipper cuts for each slide. I do all my work with glass slips so I'm ignorant here, but I would guess that coverslipping machines allow you to adjust the length of the tape used. If not, measure the length of the slip on one slide and divide the length of the roll by that. Regards - Carol ********************** Carol B. McCollough Acting Aquatic Animal Research Pathologist Oyster Disease Research Project Fisheries Service Maryland Department of Natural Resources Cooperative Oxford Laboratory 904 S. Morris Street Oxford, Maryland 21654 410-226-5193 x124 -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: Friday, December 17, 2004 10:43 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] coverslipping film[Scanned] Six sevenths of 1500, which is um........ Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Dave Johnson [mailto:djohnson14@hotmail.com] Sent: 17 December 2004 15:10 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] coverslipping film[Scanned] Eileen I have been told that the KP Tape from Mercedes Medical which is 70 meters long will cover about 1500 slides. The other tape on the market is 60 meters so i am not sure how many that covers Thanks Dave Johnson Mercedes Medical 800-331-2716 x157 >From: eileen dusek >To: histonet >Subject: [Histonet] coverslipping film >Date: Fri, 17 Dec 2004 06:27:23 -0800 (PST) > >Good Morning, >Does anyone know how many slides a roll of coverslipping film will cover. I >have to do a cost analysis for the lab. >Thanks > >Happy Holiday >Eileen >Edward Hospital > > >--------------------------------- >Do you Yahoo!? > All your favorites on one personal page - Try My Yahoo! >_______________________________________________ >Histonet mailing list >Histonet@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 Michael.Rice <@t> holy-cross.com Fri Dec 17 10:17:47 2004 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] coverslipping film[Scanned] Message-ID: <3BC92F29BE821745AB15E04C98EE028D6936CA@HCH2KMAIL.holy-cross.com> 1285 -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: Friday, December 17, 2004 10:43 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] coverslipping film[Scanned] Six sevenths of 1500, which is um........ Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Dave Johnson [mailto:djohnson14@hotmail.com] Sent: 17 December 2004 15:10 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] coverslipping film[Scanned] Eileen I have been told that the KP Tape from Mercedes Medical which is 70 meters long will cover about 1500 slides. The other tape on the market is 60 meters so i am not sure how many that covers Thanks Dave Johnson Mercedes Medical 800-331-2716 x157 >From: eileen dusek >To: histonet >Subject: [Histonet] coverslipping film >Date: Fri, 17 Dec 2004 06:27:23 -0800 (PST) > >Good Morning, >Does anyone know how many slides a roll of coverslipping film will cover. I >have to do a cost analysis for the lab. >Thanks > >Happy Holiday >Eileen >Edward Hospital > > >--------------------------------- >Do you Yahoo!? > All your favorites on one personal page - Try My Yahoo! >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- From julien_lambreydesouza <@t> uqar.qc.ca Fri Dec 17 10:55:56 2004 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Poly-NoCal Message-ID: <00b501c4e459$46c8bf00$9310d784@uqar.qc.ca> Hello all, We have a protocol for in-situ hybridization that requires Poly-Nocal for decalcification. Unfortunatly this product is discontinued. Would anybody know why the use of poly-nocal is required to any other Decal product, or would it be possible to replace it with another? Thanks, Julien Lambrey de Souza Biologie Evolutive, Universite du Quebec ? Rimouski, (418) 723-1986 #1438 From pmarcum <@t> polysciences.com Fri Dec 17 11:30:53 2004 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Poly-NoCal In-Reply-To: <00b501c4e459$46c8bf00$9310d784@uqar.qc.ca> Message-ID: <002001c4e45e$28dae770$7f00a8c0@PMARCUM2K> Poly No Cal has not been discontinued. It can not be shipped to Canada in the 3.8 liter size. Sorry I just wanted to correct this for all. Best Regards, Pamela A Marcum Histology/Microscopy Product Development Manager Polysciences, Inc. 400 Valley Road Warrington, PA 18976 Telephone: 800-523-2575???? Ext. 167 215-343-6484???? Ext. 167 Fax:? 800-343-3291 ???????? 215-343-0214 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Julien > Sent: Friday, December 17, 2004 11:56 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Poly-NoCal > > > Hello all, > > We have a protocol for in-situ hybridization that requires > Poly-Nocal for decalcification. Unfortunatly this product is > discontinued. Would anybody know why the use of poly-nocal is > required to any other Decal product, or would it be possible to > replace it with another? > > Thanks, > > Julien Lambrey de Souza > Biologie Evolutive, > Universite du Quebec ? Rimouski, > > (418) 723-1986 #1438 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Jann.Nevard <@t> baycare.org Fri Dec 17 14:23:35 2004 From: Jann.Nevard <@t> baycare.org (Nevard, Jann) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Histologist Opportunity Message-ID: <1CAFDB7DCC67D41189920008C7A44D720C64D889@SJHEXCH01> Opportunity to work in 5th largest not-for-profit health care facility nation wide and the largest on the West Coast of Florida. Currently seeking a full time histologist licensed in the state of Florida. The shift is from 3am-11:30am. Relocation and excellent benefit package. For further consideration please respond via email. Jann Nevard Regional Recruiter BayCare Heath System Ph: 727-519-1315 Toll Free: (866) 221-3222 option #1 Fax: 727-519-1323 jann.nevard@baycare.org From JCarpenter764 <@t> aol.com Fri Dec 17 15:16:16 2004 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] ok...I FINALLY GOT MY PRACTICAL!!!WOO HOO Message-ID: <46.5f0fbf9a.2ef4a6a0@aol.com> i finally got my pactical results today and passed....Just waiting on that certificate now!!! yeah....Thanks for everyones help and advice. You guys are great..Jennell From hlukey <@t> msn.com Fri Dec 17 17:48:43 2004 From: hlukey <@t> msn.com (Hugh Luk) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Kappa and Lambda Message-ID: Ellen, Kappa and Lambda staining has always been hard. It also depends on what tissues you are staining (ie liver has a lot of endogenous Biotin). What are you using as a control? Tonsil? Is your tissue fixation and processing the same as the control? How are you fixing your bone marrows? Decal solutions often changes or "kills" IHC staining parameters. Dako's Envision Plus is great (yeah, this is a very subjective statement; but tough! I'm getting good, repeatable results), but are your antibodies "Optimized" for Envision Plus? Your incubation may be too long or (if you buy concentrated antibodies) your dilution too strong. One of the hospitals I work at uses Proteinase K followed by 8 min primary, then LSAB2. My other job uses HEIR, 10 min primary made with diluent WITH Background Reducing Components (S3022), then Envision plus. Decal, because the timing changes with the pathologists/grossers, may need to be standardized. This may be the hardest part (as it was with mine, I had 23 pathologists and 9 sets of grossers). Three suggestions then: 1) Reoptimize titers to determine best time/concentration for Bouin's tissues with a Bouin's Fixed and/or decal tonsil. This may include that you "Dictate" grossing situations (fixatives, timing of fixatives, and decalcification). 2) Try a Protein block. And/or a Avidin/Biotin block. 3) Check with Dako. I have found their technical support to be awesome. As we say in Hawaii; "Eh! No be shame!" We all have had some problems with these antibodies. If Dako's Ab's are not workable with Bouin's or whatever other variables there are, ask other vendors/techs which company's antibodies work best. Good luck. Hope this helps. Hugh Luk, HTL(ASCP) hluk@crch.hawaii.edu Epidemiology Section Cancer Research Center of Hawaii 1236 Lauhala Street, Room 316 Honolulu, HI 96813 Tel: (808) 440-5238 >From: Chewy71874@aol.com >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Kappa and Lambda >Date: Fri, 17 Dec 2004 01:38:58 EST > >We are having a problem with background staining on Kappa and Lambda >stains. >We use DAKO rabbit polyclonal antibodies, DAKO TRS for antigen retrieval in >the steamer, Envision+ HRP, DAB. The background staining is especially bad >on >Bouin's fixed, decaled bone marrows. Does anyone else have this problem? >Do >you get cleaner staining with monoclonal antibodies? We are open to >suggestions. > >Thanks in advance, > >Ellen Yee, HT >Lead Tech, Immuno Dept. >Central Histology Facility >Diagnostic Pathology Medical Group >2420 J St. >Sacramento, CA 95816 >ph: 916-447-2718 >fax: 916-447-0620 >Sacramento, CA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From margnificent <@t> hotmail.com Fri Dec 17 19:31:00 2004 From: margnificent <@t> hotmail.com (Margie Teng) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Chondrocyte loss in paraffin embedded cartilage sections Message-ID: Hi, I am trying to perform immunohistochemistry assays on human cartilage sections. I have a problem with chondrocyte loss - when I look at the DAPI stains, I see a significant number of nuclei outside of the boundaries of the tissue. There are a lot of empty lacunae, although I think that is also related to the initial quality of the tissue. Does anyone have any suggestions to prevent this from happening? I am using Fisherbrand Superfrost Plus microscope slides. The assays that I have observed this with are: TUNEL, In Situ Oligo Ligation (ISOL), and an assay for single-stranded DNA using mouse monoclonal antibody. I rehydrate my samples using three changes of xylene, two changes of 100% ethanol (etOH), 1x 95% etOH, and 1x 70% etOH. I am trying to compare the difference between incubating my samples for 10 min versus 15 min in 20 microgram/mL proteinase K, but I am worried that decreasing my proteinase K incubation might mask my DNA. I can provide further details if needed. Thank you, Margie Teng ------------------------------ Margie Teng [1]tengma@orthosurg.ucsf.edu Research Assistant, Orthopaedic Surgery Veteran's Affair Medical Center, San Francisco and University (415) 221 - 4810 x 2743 References 1. mailto:tengma@orthosurg.ucsf.edu From darkdaym <@t> mindspring.com Fri Dec 17 20:05:32 2004 From: darkdaym <@t> mindspring.com (Mark Ray) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Presents for a Histo Holiday Message-ID: <41C3906C.7090502@mindspring.com> Season's Greetings All, Still trying to find something to give your Histo friends? Books are always good-they last and they don't need batteries. A couple of fine choices might be John Kiernan's _Histological and Histochemical Techniques: Theory and Practice_, ISBN 0750649364, and Kiernan and Richard Horobin's _Conn's Biological Stains_, ISBN 1859960995. We all know these guys and they know their stuff. Should be no problems about supply-the dealers have, or can easily get, all you want. _Histological and_ _Histochemical Techniques_ has been much lauded for its explication of the chemistry of routine methods and it's practical too, plenty of methodology. Any one familiar with the older editions of _Conn's_ will be greatly impressed by this one. It's organized! You can find things in it. I suggest ordering by ISBN to avoid confusion about which edition you want. You don't even need to tell the dealer or search engine the title, just the ISBN. Anyway, catch up on your reading and Happy Holidays. Mark Ray From Andrew.Gray <@t> vcp.monash.edu.au Sun Dec 19 21:44:12 2004 From: Andrew.Gray <@t> vcp.monash.edu.au (Andrew Gray) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Re: Colon whole mounts and counting aberrant crypt foci (ACF) Message-ID: <130.194.217.111.1103512155.84553@my.monash.edu.au> Hi John & Histonetters, I've done a lot of fluorescence staining on whole-mount rat ileum. The reason the tissue is pinned to the cork (or a balsawood sheet in my case) is so that it is under tension while being fixed. This produces easily-mountable flat tissue sheets. Without fixation under tension, the tissue can be quite warped, as you have seen. I expect the physical bulk of whole-mount tissues would pose a problem in keeping them adhered to slides in processing. You may be able to reduce the thickness by dissecting away unwanted layers. I would suggest you consider free-floating processing, as it is not too difficult. I use cell culture plates (96 well), with the wells containing the staining reagents. The whole-mounts are processed through the wells in sequence using a soft paint brush or fine dissecting forceps. When mounting on slides the whole-mounts are positioned while in your buffer. Then, the buffer is blotted away and an appropriate mountant is added and the slide coverslipped. This may be different for ACF staining. Best wishes, Andrew Gray PhD student Victorian College of Pharmacy, Monash University Australia From gcallis <@t> montana.edu Mon Dec 20 10:09:15 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Chondrocyte loss in paraffin embedded cartilage sections In-Reply-To: References: Message-ID: <6.0.0.22.1.20041220084821.01b5d600@gemini.msu.montana.edu> From what you describe, the chondrocytes are probably mechanically displaced from their lacunae in the cartilage during the sectioning step. Is there bone attached to the cartilage? and what size are the samples and what is your processing schedule? Cartilage tends to be drier than surrounding bone (you did not say if any was attached) due to its high water content, so during processing, more of the bound water is removed along with free water in tissue. After trimming the block, soak it on ice block with water briefly. Use a brand new disposable blade, preferably high profile to insure stability, but low profile will work IF the cartilage is perfectly processed which could entail longer schedule with thicker samples. Some disposable blades work better with cartilage/bone samples, and change to a fresh sharp edge (even between ribbons!) frequently to insure the cartilage is perfectly flat when sectioned. What you see, with nuclei askew in tissue and dried into place/adhered to slide happens before the staining. Also, if the cartilage is perfectly processed with total infiltration by paraffin - the latter will support all structures better to avoid displacement of nuclei. M 12/17/2004, you wrote: > I am trying to perform immunohistochemistry assays on human cartilage > sections. I have a problem with chondrocyte loss - when I look at the > DAPI stains, I see a significant number of nuclei outside of the > boundaries of the tissue. There are a lot of empty lacunae, although > I think that is also related to the initial quality of the tissue. > > Does anyone have any suggestions to prevent this from happening? I am > using Fisherbrand Superfrost Plus microscope slides. The assays that > I have observed this with are: TUNEL, In Situ Oligo Ligation (ISOL), > and an assay for single-stranded DNA using mouse monoclonal antibody. > > I rehydrate my samples using three changes of xylene, two changes of > 100% ethanol (etOH), 1x 95% etOH, and 1x 70% etOH. I am trying to > compare the difference between incubating my samples for 10 min versus > 15 min in 20 microgram/mL proteinase K, but I am worried that > decreasing my proteinase K incubation might mask my DNA. > > I can provide further details if needed. > > Thank you, > > Margie Teng > > ------------------------------ > Margie Teng > [1]tengma@orthosurg.ucsf.edu > Research Assistant, Orthopaedic Surgery > Veteran's Affair Medical Center, San Francisco and University > (415) 221 - 4810 x 2743 > >References > > 1. mailto:tengma@orthosurg.ucsf.edu >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From john.mcginley <@t> colostate.edu Mon Dec 20 11:42:01 2004 From: john.mcginley <@t> colostate.edu (John McGinley) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Thanks Message-ID: <200412201738.iBKHcPvP004744@neo.agsci.colostate.edu> Hi, I just wanted to say thank you for all of the responses I received regarding colon whole mounts and ACF counting. The information was quite helpful. Happy Holidays, John -------------------------- John N. McGinley Cancer Prevention Lab Colorado State University From jmahoney <@t> alegent.org Mon Dec 20 11:53:31 2004 From: jmahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Registry Message-ID: Can someone tell me what the "passing" number is for the practical part of the registry? I'd also like to know what the "perfect" score is. Thanks, Happy Holidays everyone! Jan Omaha, NE From TMcNemar <@t> lmhealth.org Mon Dec 20 12:04:51 2004 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Registry Message-ID: <6CD94D97ED7D924BA5C2B588FA952821396765@nt_exchange.lmhealth.org> Passing on the practical is a score of 400. I don't know what a perfect score would be. I've never heard of anyone getting a perfect score.... Wonder how often it happens..... Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Janice A Mahoney [mailto:jmahoney@alegent.org] Sent: Monday, December 20, 2004 12:54 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Registry Can someone tell me what the "passing" number is for the practical part of the registry? I'd also like to know what the "perfect" score is. Thanks, Happy Holidays everyone! Jan Omaha, NE _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dlcowie <@t> prodigy.net Mon Dec 20 12:28:23 2004 From: dlcowie <@t> prodigy.net (Dawn Cowie) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] histology tech job opportunity Message-ID: <20041220182823.41595.qmail@web81004.mail.yahoo.com> Happy Holidays to all! I have a full time position available for a Florida licensed histology tech. Monday thru Friday, days. Rotate one Saturday in five. We are a full service pathology lab. Duties include: embedding, microtomy, special stains, IHC. We offer a competitive salary, medical and dental benefits, generous paid time off and 9 paid holidays. Florida license as a HTL or HT required. ASCP certification preferred. Experience preferred. If interested or would like more info, please contact: Dawn Cowie, Histology Supv at 850-416-7251. Fax resume to 850-416-4471. E-mail resume to dlcowie@prodigy.net From Kristopher.Kalleberg <@t> unilever.com Mon Dec 20 12:28:44 2004 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] catalase Message-ID: I am currently working with catalase and I need to find a stain that will not be picked up in the melanin of melamocytes. I have used nuclear fast red and DAB but the red-brown color is located in both corneum and melanocytes. I need to be able to tell where exactly the catalase is alone. Thank you in advance. Kris From dlcowie <@t> prodigy.net Mon Dec 20 15:03:40 2004 From: dlcowie <@t> prodigy.net (Dawn Cowie) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Florida job opening Message-ID: <20041220210340.30735.qmail@web81010.mail.yahoo.com> hi again, Some of you asked where in Florida. Sorry, I omitted that. The position is in Pensacola Florida, located right next to the Florida/Alabama border. The company is Pensacola Pathologists. We are an independent lab located on the campus of Sacred Heart Hospital. We do all the pathology for the hospital as well as 3 other hospitals in the surrounding area. We also have a large out patient client list of our own. If anyone would like further info, please call me at 850-416-7251. Thanks, Dawn From histopath <@t> cvm.tamu.edu Mon Dec 20 18:04:03 2004 From: histopath <@t> cvm.tamu.edu (Histopath) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Registry In-Reply-To: References: Message-ID: Hi Jan, Passing score on the practical is 400. I don't know what perfect is but I received 999 on my practical. I think is has something to do with how correct the tissue blocks and slides were to what was asked for on the practical. For example, the correct sizes and the complete structures being submitted. If your sizes were 'under limit', then there might have been deductions. Then I think the readers start with this base line number and make deductions according to errors they might find in staining, fixation, microtomy, water floatation bath problems, etc. Anyway, that was the reasoning I was using. Good Luck, Rosemary Vollmar Histo Lab Supervisor Texas A&M University On Dec 20, 2004, at 11:53 AM, Janice A Mahoney wrote: > Can someone tell me what the "passing" number is for the practical > part of the registry? I'd also like to know what the "perfect" score > is. > Thanks, > Happy Holidays everyone! > Jan > Omaha, NE > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sluhisto <@t> yahoo.com Tue Dec 21 07:34:11 2004 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] OJT trained histotechs Message-ID: <20041221133411.86166.qmail@web51004.mail.yahoo.com> Hello All: With the new requirements to sit for the registry, it will be more than likely that future positions will have to be filled with BS degreed people who want to learn histology. I think that we all have figured that out. My question is this, in your institutions do you hire a "trainee" in a tech position and they sit for their registry as soon as eligible? Or, does your institution hire them in at a lower pay scale, maybe that of a lab assistant, until their training is complete and they pass the registry? As always, thanks so much for your input. Susan --------------------------------- Do you Yahoo!? Yahoo! Mail - Easier than ever with enhanced search. Learn more. From HornHV <@t> archildrens.org Tue Dec 21 08:16:23 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] OJT trained histotechs Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B9EE@EMAIL.archildrens.org> While the requirements to take the registry have changed, it doesn't mean jobs will be filled with only registered techs. While that is ideal and what we all hope for, unregistered techs will still be hired because they will be a cheap source of labor. There are no requirements that state only registered techs can do the work. Since CLIA deems the pathologist as supervisor of the histology lab as the sole source of reporting results, there is no requirement for registered techs unless your specific lab requires it. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histology SLU Sent: Tuesday, December 21, 2004 7:34 AM To: histonet@pathology.swmed.edu Subject: [Histonet] OJT trained histotechs Hello All: With the new requirements to sit for the registry, it will be more than likely that future positions will have to be filled with BS degreed people who want to learn histology. I think that we all have figured that out. My question is this, in your institutions do you hire a "trainee" in a tech position and they sit for their registry as soon as eligible? Or, does your institution hire them in at a lower pay scale, maybe that of a lab assistant, until their training is complete and they pass the registry? As always, thanks so much for your input. Susan --------------------------------- Do you Yahoo!? Yahoo! Mail - Easier than ever with enhanced search. Learn more. _______________________________________________ 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 JWEEMS <@t> sjha.org Tue Dec 21 08:57:31 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Priscilla Delventhal Message-ID: <83AACDB0810528418AA106F9AE9B7F7E507735@sjhaexc02.sjha.org> Excuse me everyone, but I need to get in touch with Priscilla. Priscilla, will you please reply? Thanks! j Happy holidays everyone. Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From Barry.R.Rittman <@t> uth.tmc.edu Tue Dec 21 09:00:05 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] OJT trained histotechs Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0010DFB43@UTHEVS3.mail.uthouston.edu> My personal opinion is that salaries of histotechs will remain below an appropriate level until there is a national policy for certification or there is unionization of histotechs. The problem is that it is difficult to trust the feds with organizing anything and second that the individual states all have their own set of criteria and seem loathe to relinquish control of these. It would be nice if all histotech certified individuals came to a job with a guaranteed level of basic training (and received a salary that was commensurate with this certification). The ASCP examination is a good first step. The written appears to test an individual's knowledge of histology and histologic technique, however I believe that the practical leaves something to be desired. It is na?ve to believe that the slides that are submitted by all individuals are all their own work. It is also not a level playing field with respect to tissue and slides. Not all individuals have the same access to tissues or facilities. The fact that slides that are submitted can be prepared using automatic staining machines makes me cringe. On the job training can be superb or it can be pitiful. I have seen individuals that fit into both these categories and between these. To me there seems to be only two solutions. First would be a national exam in which the tissues are supplies so that all candidates have the same tissues. While this is a logistical nightmare it is possible. The second is to have all job applicants sit for a simple standardized practical test at centers in each state. This also is possible. Please do not get me wrong. I believe that the ASCP exam is a good first step and is carried out as well as can be expected given the number of applicants and the current facilities. Unfortunately it is often regarded as the final step. I strongly believe that it is the responsibility of the employer to facilitate continual training of their histotechs and expect a commitment on their part want to learn and to continue with continuing education. I understand that the new requirements are for those that recently passed the histotech exam take x number CEUs over three years. I think that this is a good step. However, I feel that a person that has been in a job for 10 or 20 years, especially if in a lab with a narrow list of tissues and techniques would also greatly benefit from this. I am not sure why this is not a requirement for all histotechs. The good news is that the shortage of histotechs is making the administrators at institutes look carefully at their policies and I am hopeful that the New Year brings good news with regard to increased salaries. Hope that y'all have a really great Christmas. Hey I know we are supposed to call it winter holiday but it is really Christmas!! Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Tuesday, December 21, 2004 8:16 AM To: Histology SLU; histonet@pathology.swmed.edu Subject: RE: [Histonet] OJT trained histotechs While the requirements to take the registry have changed, it doesn't mean jobs will be filled with only registered techs. While that is ideal and what we all hope for, unregistered techs will still be hired because they will be a cheap source of labor. There are no requirements that state only registered techs can do the work. Since CLIA deems the pathologist as supervisor of the histology lab as the sole source of reporting results, there is no requirement for registered techs unless your specific lab requires it. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histology SLU Sent: Tuesday, December 21, 2004 7:34 AM To: histonet@pathology.swmed.edu Subject: [Histonet] OJT trained histotechs Hello All: With the new requirements to sit for the registry, it will be more than likely that future positions will have to be filled with BS degreed people who want to learn histology. I think that we all have figured that out. My question is this, in your institutions do you hire a "trainee" in a tech position and they sit for their registry as soon as eligible? Or, does your institution hire them in at a lower pay scale, maybe that of a lab assistant, until their training is complete and they pass the registry? As always, thanks so much for your input. Susan --------------------------------- Do you Yahoo!? Yahoo! Mail - Easier than ever with enhanced search. Learn more. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mward <@t> wfubmc.edu Tue Dec 21 09:02:44 2004 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] CD21 Message-ID: <61135F0455D33347B5AAE209B903A3040B7193BF@EXCHVS2.medctr.ad.wfubmc.edu> We are having a little problem with our CD21 and I wondered what other people are doing? Currently we are using the Dako antibody, a 1:10 dilution, Protease 2 on the NexEs. I also have a Dako instrument and Envision Plus reagents. Any help/tips would be appreciated. Martha Ward Wake Forest University Baptist Medical Center From gcallis <@t> montana.edu Tue Dec 21 10:24:51 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Happy Holidays To All! Message-ID: <6.0.0.22.1.20041221091907.01b57580@gemini.msu.montana.edu> Dear Histonetters, Happy Holidays to this great group of people who always come through for others in need of Histo information. And a special greeting to Herb Hagler and Linda Margraf - who got the (snow!)ball rolling!! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From JWEEMS <@t> sjha.org Tue Dec 21 10:32:20 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Happy Holidays To All! Message-ID: <83AACDB0810528418AA106F9AE9B7F7E50773D@sjhaexc02.sjha.org> From me too!! j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle Callis Sent: Tuesday, December 21, 2004 11:25 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Happy Holidays To All! Dear Histonetters, Happy Holidays to this great group of people who always come through for others in need of Histo information. And a special greeting to Herb Hagler and Linda Margraf - who got the (snow!)ball rolling!! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Xuesong.Chen <@t> yale.edu Tue Dec 21 10:34:26 2004 From: Xuesong.Chen <@t> yale.edu (Xuesong Chen) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] suggestions on bone cross section? Message-ID: <41C85092.9080400@yale.edu> Good morning, We are beginning a whole new approach in our lab. Basically cross sectioning undecalcified older tibias (15-19 weeks old) with cryojane type transfer system followed by lacZ staining to detect transgene expression. The tibia will be cut in the middle and proximal half close to knee joint will be used for OCT embedding. Could anybody advise how to handle the balance of the tibia vertical embedding as well as how the orientation can be precisely controlled? Also, we want to know what kind of blade is recommend for the procedure. We are wondering if anyone has any experience on bone cross sectioning and would like to offer some advise. Any input would be greatly appreciated. Thanks for you time and attention. Happy Holidays!!! Xuesong Chen Endocrinology Internal Medicine Department Yale University School of Medicine New Haven CT 06511 Tel: 203-785-2949 xuesong.chen@yale.edu From algranth <@t> u.arizona.edu Tue Dec 21 10:54:59 2004 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Happy Holidays Message-ID: <4.3.2.7.2.20041221094751.00cc22c0@algranth.inbox.email.arizona.edu> This is my last day before Christmas vacation and as I leave my warm southern red state Arizona for the cold eastern US,I also want to wish everyone a happy holiday with lots of good luck and cheer in 2005! Keep warm and be safe. Thanks to all for their thoughts, ideas and help and lively discussions. Andi Grantham ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From David.Edmondson <@t> christie-tr.nwest.nhs.uk Tue Dec 21 11:27:23 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] CD21 Message-ID: Hi, Whilst we use say 12 mins in a bath of Trypsin to digest for CD21 detection it may also be worth noting that at time we worry, our control tonsilar tissue has varying numbers of follicles and varying degrees of "exposure" though a block (maybe fixation differences). So observed staining in the control may change as one goes through a block. ( And we use Envision on a DAKO Techmate 500Plus ) That's all that comes to mind, Happy Stress-free Xmas Immuno 2U Dave Edmondson Christie Hospital Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Martha Ward Sent: 21 December 2004 15:03 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD21 We are having a little problem with our CD21 and I wondered what other people are doing? Currently we are using the Dako antibody, a 1:10 dilution, Protease 2 on the NexEs. I also have a Dako instrument and Envision Plus reagents. Any help/tips would be appreciated. Martha Ward Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ******************************************************************************************************************* This e-mail and any files transmitted with it are confidential and solely for the use of the intended recipient. If you have received this e-mail in error you should not disseminate, distribute or copy it. Please notify the sender immediately and delete this e-mail from your system. ******************************************************************************************************************* From gcallis <@t> montana.edu Tue Dec 21 11:31:06 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] suggestions on bone cross section? In-Reply-To: <41C85092.9080400@yale.edu> References: <41C85092.9080400@yale.edu> Message-ID: <6.0.0.22.1.20041221100941.01b2a9a0@gemini.msu.montana.edu> I presume mouse you are working with mouse? Are the bones prefixed? If so, you will need to sucrose cryoprotect before snap freezing. You will need a tungsten carbide knife, pricey!! and I strongly advise buying two. One in use, one being reconditioned at $220 a knife. We use D profile tungsten carbide tipped cryostat knives from Delaware Diamond Knives, and prefer these over the C profile tungsten carbide. Bones can be laid flat in an plastic embedding mold from Tissue Tek, embed inOCT, then snap freeze, preferrably with dry ice mixed with hexane. All you have to do for precise oreintation, is take frozen block, turn it on end with area you want to see facing up, and mount block on a metal cryostat chuck. Be sure to surround block with more OCT at this time to build up block face. If you mount the block with 2% methyl cellulose, you will have a very hard, sturdy support of the block but do not use this goo to embed with, it is horrible to cut. You can even stand bone on end inside a peel a way mold, the 22 mm x 22 mm. Hold the bone while OCT starts to freeze, then let it totally freeze. You now have a block that is long enough with a big enough block face to cut nicely. This amount of OCT will freeze more slowly, and you may get freezing artifact. You can freeze bones first (coated with just diluted OCT 1:1 with PBS, then double embed the frozen bone in this mold with more OCT, using dry ice/hexane slurry. You need to avoid warming up already frozen bones, use forceps that are stored in slurry to hold bone until bottom of mold begins to freeze. Be sure to release forceps before block is totally frozen. This will be easier than trying to stand a frozen bone upright on a chuck and add more OCT. We set the cryostat temperature to -29C or colder with knife and sample also that cold. Good luck as cryosectioning of tiny cortical bones will NOT be easy. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From JEllin <@t> yumaregional.org Tue Dec 21 12:07:00 2004 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] OJT trained histotechs and CLIA Message-ID: Since we are talking about the OJT I have additional food for thought. What about the CLIA regualtion categorizing histo techs. I think that it needs to be re evaluated for accuracy. Now tech are being asked to provide a higher education, computer skills, grossing and all around trouble shooting on higher level test. I do not think that the CLIA regulation hold true to what current tech are having to put out. I know that ASCP send out a questionaire about what our current duties are, but that is not even close to reflect the duties of our Techs. Anyone Idea on this?? Jesus Ellin From JEllin <@t> yumaregional.org Tue Dec 21 12:07:00 2004 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] OJT trained histotechs and CLIA Message-ID: Since we are talking about the OJT I have additional food for thought. What about the CLIA regualtion categorizing histo techs. I think that it needs to be re evaluated for accuracy. Now tech are being asked to provide a higher education, computer skills, grossing and all around trouble shooting on higher level test. I do not think that the CLIA regulation hold true to what current tech are having to put out. I know that ASCP send out a questionaire about what our current duties are, but that is not even close to reflect the duties of our Techs. Anyone Idea on this?? Jesus Ellin From info <@t> instrumedics.com Tue Dec 21 12:15:39 2004 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Happy Holidays Message-ID: <018b01c4e789$18762bf0$6401a8c0@INSTRUMEDICS22> Instrumedics wishes all Happy Holidays and a very good New Year! Bernice Schiller From danielak <@t> stanford.edu Tue Dec 21 12:31:06 2004 From: danielak <@t> stanford.edu (Daniela Kaufer) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] sending sections for immunostaining Message-ID: <7A0D01E1-537E-11D9-A476-000D9331AB4E@stanford.edu> Hello, I should receive some rat brain sections on slides from Germany, that i will then stain (IHC) here. I was wondering if anyone has suggestions as to what will be the best way to do that. We tried sending it mounted, and i opened the mounting (DEpEX) and re-hydrated them, and stained, but my staining didnt work (i know it works on fresh slides). We were next thinking of sending it in slide mailers , in PBS, but we are not sure if they stick to the slide if we keep them in solution. We can also mount them with the mounting stuff that stays liquid (I thihk it's just glycerol). I would really appreciate any advice about shipping slides. many thanks, Daniela. ==================== Daniela Kaufer, Ph.D Departments of Neurosurgery and Biological Sciences. Stanford University Stanford, CA 94305-5020 tel: 650 725-9898; fax: 650 725-5356 e-mail:danielak@stanford.edu From Edward.Crisp <@t> leica-microsystems.com Tue Dec 21 13:26:37 2004 From: Edward.Crisp <@t> leica-microsystems.com (Edward.Crisp@leica-microsystems.com) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] unsubscribe Message-ID: Edward A Crisp Segment Marketing Manager SSP Europe Leica Microsystems Davy Avenue Knowlhill Milton Keynes MK5 8LB Telephone: +44 (0)1908 246246 ext 285 Mobile: +44 (0)7786 358 649 Home Office +44 (0)115 9253 679 www.Histo-Solutions.com ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From jengirl1014 <@t> yahoo.com Tue Dec 21 13:57:17 2004 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Happy Holidays! Message-ID: <20041221195717.35014.qmail@web60605.mail.yahoo.com> May you all have a very Merry Christmas/Happy Chanuka/Happy Kwanzaa/Winter Solstice! And may next year provide you with perfect specimens, no over drying, and beautiful stains to last the whole year through! Just in case it doesn't, it's nice to know we'll all be here to help! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University Ross 929 720 Rutland Avenue Baltimore, MD 21205 phone: 410-614-0131 cell: 443-413-0853 e-mail: jengirl1014@yahoo.com --------------------------------- Do you Yahoo!? Read only the mail you want - Yahoo! Mail SpamGuard. From YPolonskaya <@t> Geron.com Tue Dec 21 14:16:14 2004 From: YPolonskaya <@t> Geron.com (Polonskaya, Yelena) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] immunohistochemestry on heart tissue Message-ID: ar colleagues, My question is for those who works with heart tissue. When I'm doing immunohistochemistry on heart tissue, I have a very high background. Sometimes, even if I don't use primary, or any antibody, I still have florescent staining. I work with perfused tissue ( 4%PFA) 20 microns. If anyone can offer a possible solution I would appreciate the help Thank you very much Yelena Phone: (650) 566-7178 e-mail: ypolonskaya@geron.com Geron cor. From frosscis <@t> usc.edu Tue Dec 21 21:21:48 2004 From: frosscis <@t> usc.edu (Fred Ross-Cisneros) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Semi-thin Epon Sectioning with wrinkles Message-ID: <001401c4e7d5$5fa1dc30$7f607d80@sadunlab> Dear All, I'm cutting cross-sectional profiles of human optic nerve processed for and embedded in epon (plastic) on a DuPont-Sorvall MT-2B ultramicrotome. The tissue is 4 mm wide (2mm thick). I am using a diamond knife for semi-thin sectioning (Histoknife by DDK) and cutting these sections usually at 0.5 microns (but I've tried cutting thinner). I'm getting lots of wrinkles in the sections and would like to know if anyone has a technique of flattening such large sections on a glass slide. I've tried chloroform vapor with no success. I have also kept the sections on the water bubble longer (glass slide on a hot plate, of course). I am aware several factors (which I have tried to control for) can contribute to wrinkling in plastic sectioning (i.e.-soft block, dirty or dull knife, cutting speed, etc., etc.). One factor I cannot get around is the large block face. I personally feel the block is quite hard (brittle). I look forward to your recommendation(s). Thank you. Fred N. Ross-Cisneros Neuro-Ophthalmology Lab USC Keck School of Medicine and Doheny Eye Institute From John.Auld <@t> whnt.nhs.uk Wed Dec 22 02:56:26 2004 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Re: CD21 Message-ID: Hi Martha Although it's been 2 -3 years since I did hands on IHC I hope this maybe of some help. In my experience CD21 is fixation dependant, the longer the tissue is fixed the longer you need to give it in enzyme. At the time I was doing them CD21 did not survive HEIR, although I beleive that option may now be available. So my suggestion is to try different times in Protease Good luck Best wishes to all for Xmas and a happy New Year John From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 22 03:08:04 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Happy Holidays[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EDCD@bhrv-nt-11.bhrv.nwest.nhs.uk> Not an advert at all......... -----Original Message----- From: Instrumedics [mailto:info@instrumedics.com] Sent: 21 December 2004 18:16 To: HistoNet Server Subject: [Histonet] Happy Holidays[Scanned] Instrumedics wishes all Happy Holidays and a very good New Year! Bernice Schiller _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From osteffe <@t> online.no Wed Dec 22 05:32:51 2004 From: osteffe <@t> online.no (Ole) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] re CD21 Message-ID: <000801c4e819$f9fde910$0100000a@brukere> Hi Martha I guess you are using the 1F8 clone form Dako? In the datasheet CD21 (1F8) its suggested to use (HIER)heat induced epitopal retrieval - TRS buffer pH 6,1 code no s1699 or s1700. I am using the s 1699 on CD21 and i think its superior to proteases. Staining intensity using proteases on eg CD21 varies much du to different fixation times in eg formaldehyde. Differences in staining intensity due to different fixation time is largly eliminated with the use of HIER and s-1699 or s-1700. I use the envision + system on this one, dilution CD21=1:30, 60min incubation roomtemp. Ole From osteffe <@t> online.no Wed Dec 22 06:32:27 2004 From: osteffe <@t> online.no (Ole) Date: Fri Sep 16 15:24:25 2005 Subject: [Histonet] Kappa and Lambda Message-ID: <001701c4e822$4d612240$0100000a@brukere> Hi Ellen I have no experience with Bouin, i will give it a shot and reply anyway. As far as i know the rabbit polyclonals K/L from DAKO are good. If you are detecting K/L in plasmacells i think many monoclonals performs equal or might be better. In cells wich express low amount of K/L (mature B-cells etc)i think your polyclonals are one of the best there is. I have some problems with K/L on bonemarrow too. Optimal staining of K/L requires optimal fixation. I think our problems on bonemarrow is due to how the sample is taken. Small biopsies often gets excessive background due to squeezing of the tissue. * Further dilution 2-3x might help. Delayed fixation might give excessive background. * Not much one can do with this?? Poor fixed samples might give heavy background staining. * Might help to reduce Antigen Retrieval with 20-35%. I have seen optimal staining in different publications can be achieved with; proteolysis, TRS antigen retrieval, Citratebuffer pH6,0, Tris-EDTA buffer pH 7,8 and combi Citratbuffer + proteolysis. I have tried most proteases, combi methods and buffers (including TRS-buffer). I found long citrate-buffer retrieval to be the best on our samples, HIER with citratebuffer pH 6,0 at 100degC about 30mins. Polyclonal K (Dako) usually somewhere around 1:4000 (dependent on incubation time/temp, and sensitivity of detection system), L usually 1,5-2x higher dilution. You might try double immunofluorescence eg L FITC / K Txr, "backgound" gets an orangecoloured staining. I use this when detecting plasmacells on eg bonemarrow with much background (with IHC-staining) im not experienced with different fluorecence-techniques, there probably better fluorochromes than Fitc and TxR. Ole From mlm11 <@t> cornell.edu Wed Dec 22 09:25:29 2004 From: mlm11 <@t> cornell.edu (Mary Lou Norman) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Semi-thin Epon Sectioning with wrinkles Message-ID: <5.2.1.1.2.20041222101927.00c5e920@postoffice9.mail.cornell.edu> Dear Fred, Try ammonia. While your sections are in the water bubble, add ammonia by dropper straight from the bottle .... .....under the hood, of course. I was impressed how flat my sections look before staining. Good luck. Happy Holidays, Mary Lou Norman Veterinary Medicine at Cornell Dear All, I'm cutting cross-sectional profiles of human optic nerve processed for and embedded in epon (plastic) on a DuPont-Sorvall MT-2B ultramicrotome. The tissue is 4 mm wide (2mm thick). I am using a diamond knife for semi-thin sectioning (Histoknife by DDK) and cutting these sections usually at 0.5 microns (but I've tried cutting thinner). I'm getting lots of wrinkles in the sections and would like to know if anyone has a technique of flattening such large sections on a glass slide. I've tried chloroform vapor with no success. I have also kept the sections on the water bubble longer (glass slide on a hot plate, of course). I am aware several factors (which I have tried to control for) can contribute to wrinkling in plastic sectioning (i.e.-soft block, dirty or dull knife, cutting speed, etc., etc.). One factor I cannot get around is the large block face. I personally feel the block is quite hard (brittle). I look forward to your recommendation(s). Thank you. Fred N. Ross-Cisneros Neuro-Ophthalmology Lab USC Keck School of Medicine and Doheny Eye Institute From Jackie.O'Connor <@t> abbott.com Wed Dec 22 10:25:14 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] CD4 CD8 in FFPE murine tissue Message-ID: Does anyone have any Christmas miracles for CD4 and CD8 in FFPE mouse? Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics From paw555 <@t> yahoo.com Wed Dec 22 10:30:35 2004 From: paw555 <@t> yahoo.com (pam plumlee) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] special stain tissue block/slide controls Message-ID: <20041222163036.57181.qmail@web11606.mail.yahoo.com> Hi all: I am developing a special stains slide and control block bank for our research pharm lab. We currently perform about 13 different stains (not including immunos). This will be a small scale project as we only do about 10-20 specials per month. Can anyone refer me to a good website or share info so I can be complete as possible on this project. We are a GLP lab. Thanks for any help and happy holidays, Pam __________________________________ Do you Yahoo!? Yahoo! Mail - Find what you need with new enhanced search. http://info.mail.yahoo.com/mail_250 From STEGTM <@t> samcstl.org Wed Dec 22 11:19:36 2004 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Slide etchers..... Message-ID: Hello all. Our hospital has agreed to let us purchase a slide etcher, now I have to research and find one. Am looking for one that will be compatible with CoPath, IE, will etch and spit out as many slides per case as recorded/billed in the accessioning and histology software. Any/nearly all suggestions will be appreciated. Thanks, Terre From gcallis <@t> montana.edu Wed Dec 22 11:23:14 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] CD4 CD8 in FFPE murine tissue In-Reply-To: References: Message-ID: <6.0.0.22.1.20041222100040.01b29730@gemini.msu.montana.edu> Dear Jackie, There are no miracles for FFPE mouse tissues and CD4/CD8 including a miracle antigen retrieval or enzyme digestion that will recover these antigens after formalin or paraformaldehyde (the enemies)! This has been discussed innumerable times on Histonet, and so far, NO ONE has come up with a solution for FFPE CD4 and CD8 success. It is an unfortunate fact of murine IHC life. You have options. If you insist on paraffin, you can only use the zinc tris buffer fixative (formalin free) which you can make up yourself. Tissues have fairly good morphology, but not equivalent to NBF fixed tissues. However, you can do CD4 and CD8, but it is advisable to use alk phos enzyme IHC since endog peroxidases are horrible, cannot be quenched after this fixation. The tissues are drier, so careful limited processing is a good idea, and lots of block face soaking before sectioning. BD Pharmingen sells the fixative, but it is really easy to make up in house. I will be happy to send method privately. Antibody concentrations are fairly close to frozen section work. I was not impressed with sectioning after this fixative and thought the tissue looked dry, so all the publications on this that tout equivalent morphology as NBF have not proved to be entirely accurate. I don't know if PLP fixation would help either, I have seen little success with this fixative. The best and other alternative is to do fresh snap frozen tissues. This is all we do with all our murine tissues, since we have to do CD4 and 8 along with several other CD markers on a single sample of tissue. This totally rules out FFPE for what we do so we don't even bother with NBF. Our morphology is excellent along with wonderful IHC staining using 75% acetone/25% absolute ethanol (A/A) fixation. You cannot substitute alcohol and you cannot do harsh endog perox block - use DAKO S2001 peroxidase block instead to save the section from bubbling off a slide. Some people use acetone, but we have found AA to be superior. If we do acetone, it is a double acetone fixation. Air dry overnight, 4C acetone 10 min, air dry 15 min in front of fan, return to 4C acetone for 10 min, air dry, rinse off OCT and proceed with staining. If you are doing large studies, there are some snap freezing methods that allow you to handle large load freezing sessions and not use liq nitrogen cooled isopentane or hexane. Method: snap freeze tissues, air dry FS overnight at RT then fix in Acetone/alcohol fixative for 5 min at RT, go directly to buffer rinse X 2. DO NOT redry sections after this fixation. Proceed with staining includine DAKO perox block, and avidin/biotin blocking. I have one method using A/A fixation, using biotinylated CD4, then Strepavidin-HRP, DAB+ from DAKO and DAB enhancer from DAKO. I had to use the glucose oxidase endogenous peroxidase block, but I had my primary antibody diluted out 1:15,000 and fading at 1:20,000. This is something you never see with FFPE, and probably not with ZnTris buffer fixative either. I suggest you use the Permanent Red (new from DAKO) as it is more sensitive (Chris van der Loos would say efficient) with the ZNTRIS fix, you probably will have even lower primary concentrations. Good luck from one of Santa's murine IHC elves At 09:25 AM 12/22/2004, you wrote: >Does anyone have any Christmas miracles for CD4 and CD8 in FFPE mouse? > >Jacqueline M. O'Connor HT(ASCP) >Abbott Laboratories >Global Pharmaceutical Research and Development >Discovery Chemotherapeutics >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Wed Dec 22 11:32:16 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] CD4 CD8 in FFPE murine tissue Message-ID: Thanks, Gayle - I've seen the previous posts re: not working on FFPE - that's why I was hoping for a Christmas miracle!! Looks like I'll have to resort to zinc. We use StreckTissueFixative in instances like this. Bah Humbug! Jackie Gayle Callis 12/22/2004 11:23 AM To: "Jackie M O'Connor" , Histonet@lists.utsouthwestern.edu cc: Subject: Re: [Histonet] CD4 CD8 in FFPE murine tissue Dear Jackie, There are no miracles for FFPE mouse tissues and CD4/CD8 including a miracle antigen retrieval or enzyme digestion that will recover these antigens after formalin or paraformaldehyde (the enemies)! This has been discussed innumerable times on Histonet, and so far, NO ONE has come up with a solution for FFPE CD4 and CD8 success. It is an unfortunate fact of murine IHC life. You have options. If you insist on paraffin, you can only use the zinc tris buffer fixative (formalin free) which you can make up yourself. Tissues have fairly good morphology, but not equivalent to NBF fixed tissues. However, you can do CD4 and CD8, but it is advisable to use alk phos enzyme IHC since endog peroxidases are horrible, cannot be quenched after this fixation. The tissues are drier, so careful limited processing is a good idea, and lots of block face soaking before sectioning. BD Pharmingen sells the fixative, but it is really easy to make up in house. I will be happy to send method privately. Antibody concentrations are fairly close to frozen section work. I was not impressed with sectioning after this fixative and thought the tissue looked dry, so all the publications on this that tout equivalent morphology as NBF have not proved to be entirely accurate. I don't know if PLP fixation would help either, I have seen little success with this fixative. The best and other alternative is to do fresh snap frozen tissues. This is all we do with all our murine tissues, since we have to do CD4 and 8 along with several other CD markers on a single sample of tissue. This totally rules out FFPE for what we do so we don't even bother with NBF. Our morphology is excellent along with wonderful IHC staining using 75% acetone/25% absolute ethanol (A/A) fixation. You cannot substitute alcohol and you cannot do harsh endog perox block - use DAKO S2001 peroxidase block instead to save the section from bubbling off a slide. Some people use acetone, but we have found AA to be superior. If we do acetone, it is a double acetone fixation. Air dry overnight, 4C acetone 10 min, air dry 15 min in front of fan, return to 4C acetone for 10 min, air dry, rinse off OCT and proceed with staining. If you are doing large studies, there are some snap freezing methods that allow you to handle large load freezing sessions and not use liq nitrogen cooled isopentane or hexane. Method: snap freeze tissues, air dry FS overnight at RT then fix in Acetone/alcohol fixative for 5 min at RT, go directly to buffer rinse X 2. DO NOT redry sections after this fixation. Proceed with staining includine DAKO perox block, and avidin/biotin blocking. I have one method using A/A fixation, using biotinylated CD4, then Strepavidin-HRP, DAB+ from DAKO and DAB enhancer from DAKO. I had to use the glucose oxidase endogenous peroxidase block, but I had my primary antibody diluted out 1:15,000 and fading at 1:20,000. This is something you never see with FFPE, and probably not with ZnTris buffer fixative either. I suggest you use the Permanent Red (new from DAKO) as it is more sensitive (Chris van der Loos would say efficient) with the ZNTRIS fix, you probably will have even lower primary concentrations. Good luck from one of Santa's murine IHC elves At 09:25 AM 12/22/2004, you wrote: >Does anyone have any Christmas miracles for CD4 and CD8 in FFPE mouse? > >Jacqueline M. O'Connor HT(ASCP) >Abbott Laboratories >Global Pharmaceutical Research and Development >Discovery Chemotherapeutics >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PBugelsk <@t> CNTUS.JNJ.COM Wed Dec 22 11:34:40 2004 From: PBugelsk <@t> CNTUS.JNJ.COM (Bugelski, Peter [CNTUS]) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Molecular Pathology Position Message-ID: Doctoral Level Research Scientist in Molecular Pathology POSITION SUMMARY: The primary role of this position is to support the John & Johnson Stem Cell Internal Venture (an operating unit within Centocor, J&J's biotechnology arm) by developing and deploying Molecular Pathology as a strategic resource. In close collaboration with the stem cell group, located in Suburban Philadelphia PA, the incumbent will design conduct and report research applying the tools of modern pathology to issues relevant to development of stem cell based therapeutics. ESSENTIAL FUNCTIONS: 1) Design, conduct and studies applying specialized laboratory techniques in Modern Pathology (e.g., immunohistochemistry, morphometry, and confocal microscopy) and Molecular Pathology (e.g., Laser Capture Microdissection, Micro-genomics, Micro-proteomics). 2) Serve as member of stem cell project teams and other teams as required. 3) In keeping with the priorities of research, to develop special expertise in the area of Molecular Pathology. 4) To present findings in oral and written form at various in-house and external fora as required. Contact: Peter J. Bugelski, Ph.D., FRCPath Director, Experimental Pathology Centocor 200 Great Valley Parkway Malvern pbugelsk@cntus.jnj.com From contact <@t> excaliburpathology.com Wed Dec 22 11:44:33 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Semi-thin Epon Sectioning with wrinkles Message-ID: <20041222174433.23108.qmail@web50303.mail.yahoo.com> Fred, Also try using a staining dish of room temperature distilled water with a couple of drops of ammonia in it to float the sections on. You might try a JB-4 plastic kit also for a softer block. Also curious why you are embedding in plastic to begin with. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From MTitford <@t> aol.com Wed Dec 22 12:07:07 2004 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Seasons Greetings from Dixie, and thanks! Message-ID: <71718325.2281F1E2.00762DB1@aol.com> Seasons greetings to everyone from the Heart of Dixie, and thanks to the Histonet organizers for their hard work. The Histonet remains a wonderful tool for the exchange of histology/technology information. Mike Titford USAPathology Mobile AL USA From Kristopher.Kalleberg <@t> unilever.com Wed Dec 22 12:15:06 2004 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] melanin bleaching Message-ID: Can someone tell me what the best way to go about bleaching melanin would be if I was trying to find label for catalase? Will the potassium permanganate destroy the antigenicity. From MadaryJ <@t> MedImmune.com Wed Dec 22 12:27:52 2004 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Need Chloral Hydrate Source Message-ID: <83899F0EC7671543B305FB5694024DE60A1B4662@medimmune4.medimmune.com> I make up Mayers and prefer Chloral Hydrate over glycerin, a little for me a little for the stain. Kidding. I am having trouble getting a source. Any ideas? I used Fisher but they claim they are not dealing drugs anymore. Nick Madary, HT/HTL(ASCP)QIHC Medimmune Histology 301.398.6113 Madaryj@medimmune.com From CMCCOLLOUGH <@t> dnr.state.md.us Wed Dec 22 12:46:20 2004 From: CMCCOLLOUGH <@t> dnr.state.md.us (McCollough, Carol) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Need Chloral Hydrate Source Message-ID: Nick: We get ours from Sigma now. C-8383. Regards - Carol ********************** Carol B. McCollough, HT/HTL(ASCP) Acting Aquatic Animal Research Pathologist Oyster Disease Research Project Fisheries Service Maryland Department of Natural Resources Cooperative Oxford Laboratory 904 S. Morris Street Oxford, Maryland 21654 410-226-5193 x124 -----Original Message----- From: Madary, Joseph [mailto:MadaryJ@MedImmune.com] Sent: Wednesday, December 22, 2004 1:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need Chloral Hydrate Source I make up Mayers and prefer Chloral Hydrate over glycerin, a little for me a little for the stain. Kidding. I am having trouble getting a source. Any ideas? I used Fisher but they claim they are not dealing drugs anymore. Nick Madary, HT/HTL(ASCP)QIHC Medimmune Histology 301.398.6113 Madaryj@medimmune.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rachael_Emerson <@t> URMC.Rochester.edu Wed Dec 22 13:18:07 2004 From: Rachael_Emerson <@t> URMC.Rochester.edu (Emerson, Rachael) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] BCIP/NBT with blocking buffer Message-ID: Hello. Can anyone tell me if I can block endogenous peroxidase with 0.3% H2O2 in sodium azide and still visualize with BCIP/NBT on frozen sections? Thanks From gcallis <@t> montana.edu Wed Dec 22 13:42:11 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] BCIP/NBT with blocking buffer In-Reply-To: References: Message-ID: <6.0.0.22.1.20041222123314.01b43248@gemini.msu.montana.edu> The answer is theroretcially - yes. However, this is not the endogenous enzyme you need to block for this chromogen. NBT/BCIP is a chromogen for alkaline phosphatase method, so blocking endogenous peroxidase is unnecessary. You need to put levamisole in with chromogen to block endogenous alkaline phosphatase found in tissues. Instruction sheets usually tell you this. Levamisole is sold separately i.e. Vector has it. OR are you doing a double stain? with both HRP and AP methods. If that is the case, you would still need to block for endog perox for the the HRP portion of staining AND block with levamisole with AP portion. At 12:18 PM 12/22/2004, you wrote: >Hello. Can anyone tell me if I can block endogenous peroxidase with 0.3% >H2O2 in sodium azide and still visualize with BCIP/NBT on frozen sections? > >Thanks > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Edward.Crisp <@t> leica-microsystems.com Wed Dec 22 19:00:47 2004 From: Edward.Crisp <@t> leica-microsystems.com (Edward.Crisp@leica-microsystems.com) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Edward Crisp/GBMIK/LMSCentral/Leica is out of the office. Message-ID: I will be out of the office starting 22/12/2004 and will not return until 03/01/2005. I will be out of the office untill Monday 3rd January, I will be able to collect my emails from wednesday 29th December, However, should you have any urgent requests please call me on my mobile. ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From juan.gutierrez <@t> christushealth.org Thu Dec 23 05:03:22 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Need Chloral Hydrate Source Message-ID: Sigma has it. Cat# C8383. 1-800-325-3010 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 Madary, Joseph Sent: Wednesday, December 22, 2004 12:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Need Chloral Hydrate Source I make up Mayers and prefer Chloral Hydrate over glycerin, a little for me a little for the stain. Kidding. I am having trouble getting a source. Any ideas? I used Fisher but they claim they are not dealing drugs anymore. Nick Madary, HT/HTL(ASCP)QIHC Medimmune Histology 301.398.6113 Madaryj@medimmune.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Dec 23 11:30:49 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] BCIP/NBT with blocking buffer In-Reply-To: References: Message-ID: <6.0.0.22.1.20041223101928.01b420a0@gemini.msu.montana.edu> The way I read your protocol, what you are detecting here is the goat serum with the rat antigoat, hence you have terrible background. In otherwords, you have an incorrect secondary antibody. If your primary is made in Rat host (rat anti-mouse) then your secondary should be goat antiRat, adsorbed to mouse, or even Donkey antiRat (Jackson). Make sure your secondary is detecting host of primary, i.e. Rat host or rat antiMouse needs either goat - or donkey antiRat adsorbed to mouse. Your normal serum block should match the host of the secondary, and use 5 to 10%. Do avidin biotin block before the primary, after normal serum block. Vector has kit, whenever you work with ABC or Strepavidin-AP method. It is strongly advisable to do a dilution panel on all your primaries starting with a target concentration of 10 ug/ml and then do a serial dilution to determine best working concentration of primary. Your secondaries should be around 2 - 5ug/ml, most of the time that means approx 1:250 dilution. Dilute your secondary with normal serum matched to host of that secondary, or just use the normal serum block. Donkey antiRat would diluted in 5% donkey serum, goat antiRat would be diluted in 5% goat serum. Handling frozen sections, cut and air dry overnight, then fix in 4C acetone 10 min, air dry for 20 min. At 12:58 PM 12/22/2004, you wrote: >Ok.....here is my basic procedure. Possibly you would have some suggestions >to help me troubleshoot. I am working with 3 different primaries. All 3 >have the same background issue. The stain is working though! > >First of all I am using frozen sections 10um thick-mouse embryos >I fix in 4 degree acetone, air dry, hydrate in PBS >Block in 1.5% normal goat serum made in 1X PBS >Primary is made in rat anti-mouse (1:200 in blocking buffer) >Secondary is rat anti-goat (1:1500 in blocking buffer) >I'm using Vector's ABC kit >Rinsing in TBST >Rinsing in NTMT with 50uL levamisole >Staining with BCIP/NBT (4.5uL NBT+3.5uL BCIP+1mL NTMT/levamisole >Rinsing in TBST > >I've also tried this with Vector Red and still had a lot of background. My >control is no primary-I substitute just blocking buffer and it has as much >background as the other slides. >I really appreciate any thoughts you may have. I am on que to restain using >a 1:400 primary concentration and blocking for 1 hour instead of 30 minutes. >Oh, I am staining for megakaryocytes. > >THANKS! >Rachael > > >Rachael L. Emerson >Center for Human Genetics and Molecular Pediatric Diseases >University of Rochester Medical Center >575 Elmwood Avenue MRBX 1.11301 >Rochester, NY 14642 > >Tel (585) 275-5073 >Fax (585) 276-0232 > > > > ---------- > > From: Gayle Callis > > Sent: Wednesday, December 22, 2004 2:42 PM > > To: Emerson, Rachael; Histonet@lists.utsouthwestern.edu > > Subject: Re: [Histonet] BCIP/NBT with blocking buffer > > > > The answer is theroretcially - yes. However, this is not the endogenous > > enzyme you need to block for this chromogen. NBT/BCIP is a chromogen for > > > > alkaline phosphatase method, so blocking endogenous peroxidase is > > unnecessary. You need to put levamisole in with chromogen to block > > endogenous alkaline phosphatase found in tissues. Instruction sheets > > usually tell you this. Levamisole is sold separately i.e. Vector has it. > > > > OR are you doing a double stain? with both HRP and AP methods. If that > > > > is the case, you would still need to block for endog perox for the the HRP > > > > portion of staining AND block with levamisole with AP portion. > > > > At 12:18 PM 12/22/2004, you wrote: > > >Hello. Can anyone tell me if I can block endogenous peroxidase with 0.3% > > >H2O2 in sodium azide and still visualize with BCIP/NBT on frozen > > sections? > > > > > >Thanks > > > > > > > > > > > >_______________________________________________ > > >Histonet mailing list > > >Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > Gayle Callis > > MT,HT,HTL(ASCP) > > Research Histopathology Supervisor > > Veterinary Molecular Biology > > Montana State University - Bozeman > > PO Box 173610 > > Bozeman MT 59717-3610 > > 406 994-6367 (lab with voice mail) > > 406 994-4303 (FAX) > > > > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Luis.Chiriboga <@t> med.nyu.edu Thu Dec 23 12:17:23 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Happy Holidays........ Message-ID: and best whishes for the new year !!!! From funderwood <@t> mcohio.org Thu Dec 23 12:17:50 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:26 2005 Subject: [BULK] - [Histonet] Happy Holidays........ Message-ID: Happy Holidays from the North Pole, I mean Dayton. It's kind of hard to tell the difference today. >>> Luis Chiriboga 12/23/04 01:17PM >>> and best whishes for the new year !!!! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From amosbrooks <@t> earthlink.net Thu Dec 23 17:41:06 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Re: Histonet Digest, Vol 13, Issue 30 In-Reply-To: <200412231001.1cHxhe1PA3NZFpr0@mx-a065a01.pas.sa.earthlink. net> References: <200412231001.1cHxhe1PA3NZFpr0@mx-a065a01.pas.sa.earthlink.net> Message-ID: <6.2.0.14.0.20041223182949.01d6cf00@pop.earthlink.net> Hi, I have found that potassium permanganate usually at least weakens a signal, so if you already have a weak signal you may have difficulty with it. The bigger problem is that sections love to fall off or get torn up when this is used. Even if you use silanized slides you are bound to experience some section loss. I wish there were a better way to remove melanin but I've not seen any that give any better results without the same problems. My advice: use low concentrations for a longer time and proceed with caution. Best of Luck, Amos Brooks At 01:01 PM 12/23/2004, you wrote: >Message: 2 >Date: Wed, 22 Dec 2004 13:15:06 -0500 >From: "Kristopher Kalleberg" >Subject: [Histonet] melanin bleaching >To: "histonet@lists.utsouthwestern.edu" > >Message-ID: >Content-Type: TEXT/PLAIN; CHARSET=US-ASCII > >Can someone tell me what the best way to go about bleaching melanin would >be if >I was trying to find label for catalase? Will the potassium permanganate >destroy the antigenicity. From lpwenk <@t> sbcglobal.net Thu Dec 23 19:39:16 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] silver proteinate Message-ID: <001701c4e959$62488ee0$24e9dc45@domainnotset.invalid> Can someone let me know a company and catalog number where I can buy silver protein/silver proteinate/protargol? It's for the Bodian. We are almost out of the container that we've had for a lot of years (company no longer in business). I've ordered twice now, and the stain does not work with what I've ordered. I know I'm doing the stain correctly, as the old protargol works great. So obviously, I'm ordering the wrong silver protein. (Yes, I know that Bielchowski is a better stain. I like to teach my students both methods, just for comparison.) Thanks. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 From jkiernan <@t> uwo.ca Fri Dec 24 00:13:57 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Re: Autofluorescence quenching References: <200412221756.iBMHu4cP009699@whipple.scsr.nevada.edu> Message-ID: <41CBB3A5.DBA9D48F@uwo.ca> Dear Daisy Narayan, Google probably referred you to in item in the "Notes and Queries" section of the journal Biotechnic & Histochemistry. The item on suppressing autofluorescence is in Vol 77 (for 2002). Biotechnic & Histochemistry is the oldest English language journal in the field of histotechnology, dating from 1925. The name was changed from Stain Technology in the early 1990s. A year's subscription to Biotechnic & Histochemistry costs about the same as 1.0ml of an antiserum (less than $200 for 6 issues). You write anonymously, but clearly from an academic institution in the USA (... unr.edu). If your library no longer takes B&H please persuade them to renew their subscription, at least for the on-line (web) version. This comes as part of a large package of journals from the publisher Taylor & Francis. You may well have access already, without knowing it. Check out http://www.tandf.co.uk/journals/ You should be able to get the full text of the item if your library subscribes. If you cannot find the "suppressing autofluorescence" item, ask your library to subscribe. Or take out a lab subscription to B&H. Nearly every issue of B&H contains something to interest everyone who works with tissues and microscopes, in a wide range of fields. If this seems like a sales pitch, it isn't. I have had a personal subscription to the journal since the 1970s (when the sub was about $50) and have had good value for money every year. Relative to inflation and cost of living, B&H is cheaper now than in earlier years. The quality has improved. Nearly all staining-related papers in B&H now have coloured illustrations. If you have difficulty obtaining the autofluorescence item, let me know. I can send you a .pdf file of that set of Notes & Queries. It includes anecdotal tips and references for various strategies. There is no way to cope with autofluorescence problems without using a big textbook (Pearse is the only big one in English) or an afternoon in a library, or both. Don't believe anything you read on the Web about autofluorescence until you have checked references to papers in peer reviewed journals. Biotechnic & Histochemistry is a peer-reviewed journal. Another, with even higher standards, is the Journal of Histochemistry and Cytochemistry. This is also inexpensive and it comes out every month. Both journals are published by non-profit societies. Any lab can subscribe to both for less than $400 a year. John Kiernan London, Canada. From juan.gutierrez <@t> christushealth.org Fri Dec 24 05:13:25 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] silver proteinate Message-ID: We use to get ours from Kodak, but I don't know if they still sell it. Good luck and Happy Holidays! 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 lpwenk@sbcglobal.net Sent: Thursday, December 23, 2004 7:39 PM To: Histonet Subject: [Histonet] silver proteinate Can someone let me know a company and catalog number where I can buy silver protein/silver proteinate/protargol? It's for the Bodian. We are almost out of the container that we've had for a lot of years (company no longer in business). I've ordered twice now, and the stain does not work with what I've ordered. I know I'm doing the stain correctly, as the old protargol works great. So obviously, I'm ordering the wrong silver protein. (Yes, I know that Bielchowski is a better stain. I like to teach my students both methods, just for comparison.) Thanks. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Fri Dec 24 05:46:36 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] silver proteinate Message-ID: I found a supplier for you. Merck has a Bodian procedure and the cat# for the protargol is: 1.07447 Silver Protein. www.gold-lustre-pigments.com/servlet/PB/show/1288850/107447en.pdf This link should bring up the reference to their procedure. 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 GUTIERREZ, JUAN Sent: Friday, December 24, 2004 5:13 AM To: lpwenk@sbcglobal.net; Histonet Subject: RE: [Histonet] silver proteinate We use to get ours from Kodak, but I don't know if they still sell it. Good luck and Happy Holidays! 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 lpwenk@sbcglobal.net Sent: Thursday, December 23, 2004 7:39 PM To: Histonet Subject: [Histonet] silver proteinate Can someone let me know a company and catalog number where I can buy silver protein/silver proteinate/protargol? It's for the Bodian. We are almost out of the container that we've had for a lot of years (company no longer in business). I've ordered twice now, and the stain does not work with what I've ordered. I know I'm doing the stain correctly, as the old protargol works great. So obviously, I'm ordering the wrong silver protein. (Yes, I know that Bielchowski is a better stain. I like to teach my students both methods, just for comparison.) Thanks. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 _______________________________________________ Histonet mailing list Histonet@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 bruyntjes <@t> voeding.tno.nl Mon Dec 27 04:45:57 2004 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] catalase Message-ID: <3B070848E7C2204F9DEB8BCFD767728004A69466@ntexch1.voeding.tno.nl> Kris I've had the same kind of problem working with DAB on skin slides, but then I changed to another chromogen called NovaRed. The color is brick red and differentiates very well between the NovaRed and the melanine cells. I ordered this NovaRed from Vector. Hopes this will help you a bit. Joost Bruijntjes TNO Nutrition and Food Research PO Box 360 3700 AJ Zeist The Netherlands -----Oorspronkelijk bericht----- Van: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Namens Kristopher Kalleberg Verzonden: maandag 20 december 2004 19:29 Aan: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] catalase I am currently working with catalase and I need to find a stain that will not be picked up in the melanin of melamocytes. I have used nuclear fast red and DAB but the red-brown color is located in both corneum and melanocytes. I need to be able to tell where exactly the catalase is alone. Thank you in advance. Kris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Matthew_Frank <@t> URMC.Rochester.edu Mon Dec 27 11:18:54 2004 From: Matthew_Frank <@t> URMC.Rochester.edu (Frank, Matthew) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] silver proteinate Message-ID: Try Polysciences in Warrington PA. cat #01070. The website is www.polysciences.com . They have protargol S that is BSC certified. -----Original Message----- From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] Sent: Thursday, December 23, 2004 8:39 PM To: Histonet Subject: [Histonet] silver proteinate Can someone let me know a company and catalog number where I can buy silver protein/silver proteinate/protargol? It's for the Bodian. We are almost out of the container that we've had for a lot of years (company no longer in business). I've ordered twice now, and the stain does not work with what I've ordered. I know I'm doing the stain correctly, as the old protargol works great. So obviously, I'm ordering the wrong silver protein. (Yes, I know that Bielchowski is a better stain. I like to teach my students both methods, just for comparison.) Thanks. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From conniegrubaugh <@t> hotmail.com Mon Dec 27 12:25:17 2004 From: conniegrubaugh <@t> hotmail.com (connie grubaugh) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Histotechnologist Position in Las Vegas, Nv Message-ID: Position available in outpatient pathology laboratory to perform routine histology procedures and gross examination of Dermatopathology specimens. Qualifications include BA/BS degree in a biological science, and HTL (ASCP) certification. Competitive compensation and benefits, no state income tax. Please fax, mail or email resume to: Karen Hackworth, Operations Manager 3059 South Maryland Parkway #100 Las Vegas Nevada 89109-2201 Phone 702-938-9911 Fax 702-732-2310 email: hackworth@lasvegaspathologist.com From Joanholtz <@t> aol.com Mon Dec 27 15:49:00 2004 From: Joanholtz <@t> aol.com (Joanholtz@aol.com) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] please advise: bor practical Message-ID: <8a.1cff89bc.2f01dd4c@aol.com> The hospital in which I work uses manual processes for all of our cutting and special staining needs. The only automation we use is the H&E. I would think this is the preferred way to learn hands on and was quite proud to learn this way. I was assured that after passing the written portion of the exam, (which I did on my first attempt) that the practical should be no problem at all. I have submitted my practical twice; this time I am eligible to submit my slides for a reevaluation along with more money. Before each submission my slides were reviewed by at least two of our pathologists. If 800 is the optimal score I do not think it is reasonable to assume that my hospitals lab puts out or accepts anything below a 300. I would truly appreciate any and all help or insights as I try and appeal my scores by Jan. 10th 2005. Am I at a disadvantage without automation? How should I select slides to be reevaluated? Is it possible to get a worse score? Am I wasting my time? Is the evaluation process anonymous, do they know your background? This is truly a very frustrating situation with a BS in Biology and three years histology experience. I have invested a lot of time, energy and money into getting where I am now and this arbitrary process is preventing me from going any further. Thank you for any and all help, Joan From mrsgbd2001 <@t> yahoo.com Mon Dec 27 16:48:32 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] please advise: bor practical In-Reply-To: <8a.1cff89bc.2f01dd4c@aol.com> Message-ID: <20041227224832.28750.qmail@web52701.mail.yahoo.com> Hey Joan, I'm in the exact same boat as you. Our lab does all work manual except the H&E stain. There were seven of us, from my lab, taking the test this fall. Out of the seven I was the only one to fail. (I am also sending my slides in for review.) I don't see how I could have failed - based on the staining of three of my H&E slides - when I used the same H&E machine my co-workers used. Not that a degree gives a person more "talent", but I am the only one of the seven with a B.S., and I wonder if that they scored me on a higher scale - which would not seem fair. I was told that the evaluation process was anonymous, by a member of NSH. I would also be interested in what happens with the reevaluation process. Good luck to you. Gareth --- Joanholtz@aol.com wrote: > The hospital in which I work uses manual processes > for all of our cutting and > special staining needs. The only automation we use > is the H&E. I would > think this is the preferred way to learn hands on > and was quite proud to learn > this way. I was assured that after passing the > written portion of the exam, > (which I did on my first attempt) that the practical > should be no problem at all. > I have submitted my practical twice; this time I am > eligible to submit my > slides for a reevaluation along with more money. > Before each submission my > slides were reviewed by at least two of our > pathologists. If 800 is the optimal > score I do not think it is reasonable to assume that > my hospitals lab puts out > or accepts anything below a 300. I would truly > appreciate any and all help or > insights as I try and appeal my scores by Jan. 10th > 2005. > > Am I at a disadvantage without automation? How > should I select slides to be > reevaluated? Is it possible to get a worse score? > Am I wasting my time? Is > the evaluation process anonymous, do they know your > background? > > This is truly a very frustrating situation with a BS > in Biology and three > years histology experience. I have invested a lot of > time, energy and money into > getting where I am now and this arbitrary process is > preventing me from going > any further. > > Thank you for any and all help, > Joan > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________ Do you Yahoo!? Yahoo! Mail - You care about security. So do we. http://promotions.yahoo.com/new_mail From malek.j <@t> ghc.org Mon Dec 27 17:03:27 2004 From: malek.j <@t> ghc.org (Malek, Jack M) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Blocks for Send Out Studies Message-ID: <63EB2FB8C50AD242898B76E2B2C7C12202121A14@ROC2T9.ghc.org> Hi Everyone, Recently learned that some pathology labs send out slide sections in place of blocks for reference studies. They withhold the blocks for obvious reasons such as; to prevent losing tissue in transport, to determine which studies to exhaust the block for, preventing laser destruction of the block associated with certain molecular studies. Is this becoming the standard in practice? Happy Holidays, Jack GHC -Seattle From lpwenk <@t> sbcglobal.net Tue Dec 28 08:46:37 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] bor practical Message-ID: <000d01c4ecec$0aa2e0a0$ea2ed445@domainnotset.invalid> Information in general about the ASCP practicals: When someone does not pass the practical exam, they are given a list of what problems were found during the grading session. The criteria for passing are sent to the candidates in the practical booklet with the list of tissues needed, and are also published on the ASCP Board of Registry webpage. http://www.ascp.org/bor/getcertified/index.asp Click on HT or HTL, then click on Practical Exam Instructions. The list of tissues may be different from session to session, but the list of grading criteria (IV) is always the same. These criteria are the same, regardless if the - processing is by hand or on an enclosed processor - sectioned by a rotary microtome or an electric model - stained by hand or on machine, either H&E or special stains - coverslipping is by hand or by machine The histotech is responsible for the QUALITY of the final slide, regardless of the method. So, look at the list of problems, and see if there is a pattern. - Was the correct tissue collected - small intestine, not large intestine, for example? - Was the minimum size of the tissue correct, and contain all requirements (kidney with cortex and medulla, for example, or gallbladder containing epithelium across one entire surface)? - Was processing complete, or was there over or under processing? - Was the tissue orientation correct during embedding - complete cross-section of fallopian tube, for example? Skin embedded on edge to see all layers? - Were there knife lines in all the sections? Or microchatter? Or bubbles/holes/tears in the tissue? Too thick? - Was there nuclear bubbling? Formalin pigment? Autolysis? - Was labeling correct on the block and/or slide? - Was there 2 tone eosin (cytoplasmic differentiation inadequate), or contrast inadequate (nuclear or cytoplasmic staining too light or dark)? - Was desired structure for the special stains understained or overstained, or was counterstain too dark? - Was the correct special stain done - PAS-Light Green, for example, not PASH? - Were there bubbles under the coverslips? Use this list of problems to improve next time. Peggy A. Wenk, HTL(ASCP)SLS Program Director, Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 From lpwenk <@t> sbcglobal.net Tue Dec 28 08:54:26 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] please advise: bor practical References: <8a.1cff89bc.2f01dd4c@aol.com> Message-ID: <001e01c4eced$20b73b60$ea2ed445@domainnotset.invalid> See my other email for more details. Just wanted to comment - 800 is not "the optimal score". 400 is the minimum pass score established by the ASCP Board of Registry Histology Exam committee, made up of histotechs and pathologists (both human and veterinary techs and doctors). The range printed out on the computer scores is 100-999, though in theory the scores can go lower than 100 and higher than 999. But the computer can only print out 3 digit numbers, and cannot print out lower than 100. Very few people get up in the 900's. In the January - June 2004 exams, the following is the ranges of the practical exams: HT = 189 - 889 HTL = 328 - 712 ftp://ftp.ascp.org/Grab/PDFs/BORpdf/Fall2004.pdf >From the ASCP BOR Newsletter, Fall 2004 Peggy A. Wenk, HTL(ASCP)SLS Program Directors, Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Monday, December 27, 2004 4:49 PM Subject: [Histonet] please advise: bor practical > The hospital in which I work uses manual processes for all of our cutting and > special staining needs. The only automation we use is the H&E. I would > think this is the preferred way to learn hands on and was quite proud to learn > this way. I was assured that after passing the written portion of the exam, > (which I did on my first attempt) that the practical should be no problem at all. > I have submitted my practical twice; this time I am eligible to submit my > slides for a reevaluation along with more money. Before each submission my > slides were reviewed by at least two of our pathologists. If 800 is the optimal > score I do not think it is reasonable to assume that my hospitals lab puts out > or accepts anything below a 300. I would truly appreciate any and all help or > insights as I try and appeal my scores by Jan. 10th 2005. > > Am I at a disadvantage without automation? How should I select slides to be > reevaluated? Is it possible to get a worse score? Am I wasting my time? Is > the evaluation process anonymous, do they know your background? > > This is truly a very frustrating situation with a BS in Biology and three > years histology experience. I have invested a lot of time, energy and money into > getting where I am now and this arbitrary process is preventing me from going > any further. > > Thank you for any and all help, > Joan > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Tue Dec 28 09:04:52 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] please advise: bor practical References: <20041227224832.28750.qmail@web52701.mail.yahoo.com> Message-ID: <003001c4ecee$95b2a160$ea2ed445@domainnotset.invalid> Just wondering - Did you take the HTL exam and the rest of the people take the HT exam? Since you have a BS? The HTL exam IS harder than the HT exam - larger tissues sections, trickier stains, often needing to cut thinner tissues, tissues that are more difficult to obtain, etc. I train both HT and HTL students, and can attest that the HTL exam is harder to do than the HT exam. Peggy A. Wenk, HTL(ASCP)SLS Program Director, Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Gareth Davis" To: ; "Histonet" Sent: Monday, December 27, 2004 5:48 PM Subject: Re: [Histonet] please advise: bor practical > Hey Joan, > I'm in the exact same boat as you. Our lab does all > work manual except the H&E stain. There were seven of > us, from my lab, taking the test this fall. Out of the > seven I was the only one to fail. (I am also sending > my slides in for review.) I don't see how I could > have failed - based on the staining of three of my H&E > slides - when I used the same H&E machine my > co-workers used. Not that a degree gives a person > more "talent", but I am the only one of the seven with > a B.S., and I wonder if that they scored me on a > higher scale - which would not seem fair. I was told > that the evaluation process was anonymous, by a member > of NSH. > I would also be interested in what happens with the > reevaluation process. > > Good luck to you. > Gareth > > --- Joanholtz@aol.com wrote: > > > The hospital in which I work uses manual processes > > for all of our cutting and > > special staining needs. The only automation we use > > is the H&E. I would > > think this is the preferred way to learn hands on > > and was quite proud to learn > > this way. I was assured that after passing the > > written portion of the exam, > > (which I did on my first attempt) that the practical > > should be no problem at all. > > I have submitted my practical twice; this time I am > > eligible to submit my > > slides for a reevaluation along with more money. > > Before each submission my > > slides were reviewed by at least two of our > > pathologists. If 800 is the optimal > > score I do not think it is reasonable to assume that > > my hospitals lab puts out > > or accepts anything below a 300. I would truly > > appreciate any and all help or > > insights as I try and appeal my scores by Jan. 10th > > 2005. > > > > Am I at a disadvantage without automation? How > > should I select slides to be > > reevaluated? Is it possible to get a worse score? > > Am I wasting my time? Is > > the evaluation process anonymous, do they know your > > background? > > > > This is truly a very frustrating situation with a BS > > in Biology and three > > years histology experience. I have invested a lot of > > time, energy and money into > > getting where I am now and this arbitrary process is > > preventing me from going > > any further. > > > > Thank you for any and all help, > > Joan > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - You care about security. So do we. > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Tue Dec 28 09:08:06 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] please advise: bor practical References: <20041227224832.28750.qmail@web52701.mail.yahoo.com> Message-ID: <003501c4ecef$09870900$ea2ed445@domainnotset.invalid> One other comment - The tech evaluators are ASCP certified HT and HTL, who are also NSH members. Some of the evaluators are pathologists, either human or veterinary. Since the candidate's social security number is the only identification on the slides and labels, the grading is anonymous. The graders do not know the candidate's name, age, sex, lab, state, etc. Peggy A. Wenk, HTL(ASCP)SLS Program Director, Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Gareth Davis" To: ; "Histonet" Sent: Monday, December 27, 2004 5:48 PM Subject: Re: [Histonet] please advise: bor practical > Hey Joan, > I'm in the exact same boat as you. Our lab does all > work manual except the H&E stain. There were seven of > us, from my lab, taking the test this fall. Out of the > seven I was the only one to fail. (I am also sending > my slides in for review.) I don't see how I could > have failed - based on the staining of three of my H&E > slides - when I used the same H&E machine my > co-workers used. Not that a degree gives a person > more "talent", but I am the only one of the seven with > a B.S., and I wonder if that they scored me on a > higher scale - which would not seem fair. I was told > that the evaluation process was anonymous, by a member > of NSH. > I would also be interested in what happens with the > reevaluation process. > > Good luck to you. > Gareth > > --- Joanholtz@aol.com wrote: > > > The hospital in which I work uses manual processes > > for all of our cutting and > > special staining needs. The only automation we use > > is the H&E. I would > > think this is the preferred way to learn hands on > > and was quite proud to learn > > this way. I was assured that after passing the > > written portion of the exam, > > (which I did on my first attempt) that the practical > > should be no problem at all. > > I have submitted my practical twice; this time I am > > eligible to submit my > > slides for a reevaluation along with more money. > > Before each submission my > > slides were reviewed by at least two of our > > pathologists. If 800 is the optimal > > score I do not think it is reasonable to assume that > > my hospitals lab puts out > > or accepts anything below a 300. I would truly > > appreciate any and all help or > > insights as I try and appeal my scores by Jan. 10th > > 2005. > > > > Am I at a disadvantage without automation? How > > should I select slides to be > > reevaluated? Is it possible to get a worse score? > > Am I wasting my time? Is > > the evaluation process anonymous, do they know your > > background? > > > > This is truly a very frustrating situation with a BS > > in Biology and three > > years histology experience. I have invested a lot of > > time, energy and money into > > getting where I am now and this arbitrary process is > > preventing me from going > > any further. > > > > Thank you for any and all help, > > Joan > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - You care about security. So do we. > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GauchV <@t> mail.amc.edu Tue Dec 28 09:12:35 2004 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Blocks for Send Out Studies Message-ID: Jack, I can only tell you that we do not release our paraffin blocks either. We offer unstained slides or 25 micron sections in a test tube,etc.- anything we can reasonably do to help them out but the blocks remain in our custody. We started this several years ago because we had issues with blocks not being returned upon completion of studies or being lost in shipping to us. There have been very few times when we have not been able to fill a request for the requesting institutions using our current system. Vicki Gauch Albany Medical Center Albany, NY >>> "Malek, Jack M" 12/27/2004 6:03:27 PM >>> Hi Everyone, Recently learned that some pathology labs send out slide sections in place of blocks for reference studies. They withhold the blocks for obvious reasons such as; to prevent losing tissue in transport, to determine which studies to exhaust the block for, preventing laser destruction of the block associated with certain molecular studies. Is this becoming the standard in practice? Happy Holidays, Jack GHC -Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akbitting <@t> geisinger.edu Tue Dec 28 09:20:17 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Cutting using (Surgipath) Supercassettes Message-ID: Has anyone heard of a Johns-Hopkins clamp? Supposedly, can get these for a R-J 2030 microtome to cut using Supercassettes. I thought the clamp I have is this type, but it's not, so I have to find one soon. Thanks folks! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From osteffe <@t> online.no Tue Dec 28 09:29:34 2004 From: osteffe <@t> online.no (Ole) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Sevier Munger Silver stain Message-ID: <000e01c4ecf2$0a2c9070$0100000a@brukere> Hi I need a protocol/receipe of the Sevier Munger silver stain. I have not found it in any of my histotech books nor had any luck searching the web. I work with PCR, immunohistochemistry and in situs and dont know much about special stains, or where to find them, if their not in our "special stain" books. It is to be used on Formaldehyd fixed paraffin embedded tissue. Best regards Ole From mrsgbd2001 <@t> yahoo.com Tue Dec 28 09:36:46 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] please advise: bor practical In-Reply-To: <003001c4ecee$95b2a160$ea2ed445@domainnotset.invalid> Message-ID: <20041228153646.66909.qmail@web52705.mail.yahoo.com> thanks for information on the HT Practical. I did realize that there were other criteria involved in the grading, but most of my comments were on staining. Anyway, I thought about taking the HTL, but it seems most people I know only have an HT and are doing almost as well in the field. Thanks again, Gareth Davis lpwenk@sbcglobal.net wrote: Just wondering - Did you take the HTL exam and the rest of the people take the HT exam? Since you have a BS? The HTL exam IS harder than the HT exam - larger tissues sections, trickier stains, often needing to cut thinner tissues, tissues that are more difficult to obtain, etc. I train both HT and HTL students, and can attest that the HTL exam is harder to do than the HT exam. Peggy A. Wenk, HTL(ASCP)SLS Program Director, Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Gareth Davis" To: ; "Histonet" Sent: Monday, December 27, 2004 5:48 PM Subject: Re: [Histonet] please advise: bor practical > Hey Joan, > I'm in the exact same boat as you. Our lab does all > work manual except the H&E stain. There were seven of > us, from my lab, taking the test this fall. Out of the > seven I was the only one to fail. (I am also sending > my slides in for review.) I don't see how I could > have failed - based on the staining of three of my H&E > slides - when I used the same H&E machine my > co-workers used. Not that a degree gives a person > more "talent", but I am the only one of the seven with > a B.S., and I wonder if that they scored me on a > higher scale - which would not seem fair. I was told > that the evaluation process was anonymous, by a member > of NSH. > I would also be interested in what happens with the > reevaluation process. > > Good luck to you. > Gareth > > --- Joanholtz@aol.com wrote: > > > The hospital in which I work uses manual processes > > for all of our cutting and > > special staining needs. The only automation we use > > is the H&E. I would > > think this is the preferred way to learn hands on > > and was quite proud to learn > > this way. I was assured that after passing the > > written portion of the exam, > > (which I did on my first attempt) that the practical > > should be no problem at all. > > I have submitted my practical twice; this time I am > > eligible to submit my > > slides for a reevaluation along with more money. > > Before each submission my > > slides were reviewed by at least two of our > > pathologists. If 800 is the optimal > > score I do not think it is reasonable to assume that > > my hospitals lab puts out > > or accepts anything below a 300. I would truly > > appreciate any and all help or > > insights as I try and appeal my scores by Jan. 10th > > 2005. > > > > Am I at a disadvantage without automation? How > > should I select slides to be > > reevaluated? Is it possible to get a worse score? > > Am I wasting my time? Is > > the evaluation process anonymous, do they know your > > background? > > > > This is truly a very frustrating situation with a BS > > in Biology and three > > years histology experience. I have invested a lot of > > time, energy and money into > > getting where I am now and this arbitrary process is > > preventing me from going > > any further. > > > > Thank you for any and all help, > > Joan > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - You care about security. So do we. > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From vazquezr <@t> ohsu.edu Tue Dec 28 09:41:30 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Sevier Munger Silver stain Message-ID: Ole, Do you have fax no.? I have recipe for Sevier-Munger Method for neural tissues Robyn Ohsu >>> "Ole" 12/28/2004 7:29:34 AM >>> Hi I need a protocol/receipe of the Sevier Munger silver stain. I have not found it in any of my histotech books nor had any luck searching the web. I work with PCR, immunohistochemistry and in situs and dont know much about special stains, or where to find them, if their not in our "special stain" books. It is to be used on Formaldehyd fixed paraffin embedded tissue. Best regards Ole _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mlfiguei <@t> ucla.edu Mon Dec 27 12:44:38 2004 From: mlfiguei <@t> ucla.edu (FIGUEIREDO,MARXA L ) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] help In-Reply-To: <200412271805.iBRI5qjs022146@smtp.ucla.edu> References: <200412271805.iBRI5qjs022146@smtp.ucla.edu> Message-ID: <1104173078.41d05816c1428@mail.ucla.edu> Hi HIstonetters, Has anyone done imunohistochemisty with anti-GFP in paraffin tissues recently? I have the anti-GFP monoclonal from Clontech (JL-8) and was wondering if anyone has tried that one in FFPE tissues. If so, what was the dilution and antigen retrieval method? THanks Marxa Figueiredo UCLA Dentistry From juan.gutierrez <@t> christushealth.org Tue Dec 28 09:54:54 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Sevier Munger Silver stain Message-ID: It's in the AFIP manual. The reference is; Sevier, A. C., and Munger, B. L.: J. Neuropath. Exp. Neurol. 24:130-135, 1965. Sorry but I don't have time to type the whole thing. 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 Ole Sent: Tuesday, December 28, 2004 9:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sevier Munger Silver stain Hi I need a protocol/receipe of the Sevier Munger silver stain. I have not found it in any of my histotech books nor had any luck searching the web. I work with PCR, immunohistochemistry and in situs and dont know much about special stains, or where to find them, if their not in our "special stain" books. It is to be used on Formaldehyd fixed paraffin embedded tissue. Best regards Ole _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MTitford <@t> aol.com Tue Dec 28 10:44:28 2004 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] On passing BOR exam Message-ID: <3F49D0EC.73796666.00762DB1@aol.com> Joan Holtz asks about passing the BOR exam. I was shocked a few years ago when I read the ASCP newsletter that gave statistics for the registry exams. Many histotechs failed the HT and HTL practicals. Now, not talking about Joans laboratory or anyone else's in particular, I suspect many applicants bang out their exam slides like they do their daily work slides and send them off without critically examining them first. In our laboratory we receive slides for consultation which are sub-standard. Obviously those laboratories think they are acceptable. With HT and HTL practical exam slides you should produce the best d*** slides you can! You have plenty of time. Scrounge for the best fixed tissues. Submit tissues of right size allowing for shrinkage. Process correctly. Cut correctly. Produce best stained slides you can. In histology, it is easy to produce adequate slides, but hard to produce perfect slides.The slides should be of right thickness, no scores, folds, precipitates, background staining etc. The staining should be perfect! Remember, if you submit a poorly stained slide, you are telling the examiner thats you best work. In my laboratory when preparing for practical exams, I expect histotechs to make multiple blocks, and multiple stained slides from those blocks. We sit down together several times and look closely at those slides. There is much going back and re-submitting tissue. We have only had one histotech fail the practical in 25 years (And that tech did not want me to see those slides!) Mike Titford USA Pathology Mobile AL USA From MTitford <@t> aol.com Tue Dec 28 10:59:02 2004 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Argyrophil stains. Message-ID: <49AF28FD.5B6B2BA3.00762DB1@aol.com> Ole, somewhere overseas I suspect, asks about the Sevier Munger stain. The reference is: Sevier, A.C., Munger, B.L., A silver method for paraffin sections of neural tissue. J. Neuropath Exp Neurol. 24: 130 - 135 1965 However, in the USA (and is it not interesting how labs use procedures written by their own countrymen?)many labs use the Churukian method which demonstrates a wider variety of argyrophil tumors. Churukian, J.,Schenk, E.A. A modification of Pascual's argyrophil method. J. Histotechnol 3: 102 - 102 1979. I read somewhere, long ago, that there is no one method which will demonstrate ALL argyophil tumors, but the Chrukian method demonstrates the widest variety. Mike Titford USA Pathology Mobile AL USA From cfavara <@t> niaid.nih.gov Tue Dec 28 11:32:48 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] help Message-ID: I have been doing anti GFP in mouse FFPE tissue. I am presently using Molecular Probes Rabbit polyclonal 2mg/ml at a dilution of 1:500 using a biotinylated secondary from Vector @1:250 dil on the Ventana Nexus. I use Citrate pH 6.0 in a pressure cooker temp 120 for 30 seconds. I will be evaluating monoclonal from Zymed and BD Biosciences when I get the time. Sorry I have no experience with Clontech. 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: FIGUEIREDO,MARXA L [mailto:mlfiguei@ucla.edu] Sent: Monday, December 27, 2004 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] help Hi HIstonetters, Has anyone done imunohistochemisty with anti-GFP in paraffin tissues recently? I have the anti-GFP monoclonal from Clontech (JL-8) and was wondering if anyone has tried that one in FFPE tissues. If so, what was the dilution and antigen retrieval method? THanks Marxa Figueiredo UCLA Dentistry _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Charles.Embrey <@t> carle.com Tue Dec 28 11:49:32 2004 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] On passing BOR exam Message-ID: Thank-you! Thank-you! Thank-you! I am shocked by the laid-back attitude that many people today take in approaching the practical exam. They don't seem to even read the grading criteria required to produce the slides capable of passing. Each slide sent in to be graded should first be graded by the technician against these grading guidelines. There should really be no surprises. If you don't know enough about histology to grade your own slides you really shouldn't be certified as a histotech. As far as automation goes I think all practical exam slides should be produced WITHOUT automation. I don't ever want to hire a histotech that can't do a GMS by hand. Sorry if this seems harsh but as a lab manager I want to see the BOR stamp of approval mean something. Quit wringing your hands; take a look at the list of errors provided by the grader and produce slides that avoid those mistakes. Charles Embrey, Pathologists' Assistant, HT(ASCP) Histology Manager Carle Clinic, Urbana Illinois -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of MTitford@aol.com Sent: Tuesday, December 28, 2004 10:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] On passing BOR exam Joan Holtz asks about passing the BOR exam. I was shocked a few years ago when I read the ASCP newsletter that gave statistics for the registry exams. Many histotechs failed the HT and HTL practicals. Now, not talking about Joans laboratory or anyone else's in particular, I suspect many applicants bang out their exam slides like they do their daily work slides and send them off without critically examining them first. In our laboratory we receive slides for consultation which are sub-standard. Obviously those laboratories think they are acceptable. With HT and HTL practical exam slides you should produce the best d*** slides you can! You have plenty of time. Scrounge for the best fixed tissues. Submit tissues of right size allowing for shrinkage. Process correctly. Cut correctly. Produce best stained slides you can. In histology, it is easy to produce adequate slides, but hard to produce perfect slides.The slides should be of right thickness, no scores, folds, precipitates, background staining etc. The staining should be perfect! Remember, if you submit a poorly stained slide, you are telling the examiner thats you best work. In my laboratory when preparing for practical exams, I expect histotechs to make multiple blocks, and multiple stained slides from those blocks. We sit down together several times and look closely at those slides. There is much going back and re-submitting tissue. We have only had one histotech fail the practical in 25 years (And that tech did not want me to see those slides!) Mike Titford USA Pathology Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Lynne.Bell <@t> hitchcock.org Tue Dec 28 12:23:16 2004 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Fetal Remains Message-ID: I am interested how other hospital handle and/or dispose of fetal remains. In particular, I am interested in "products of conception" where there is an actual formed fetus. How long do you retain this specimen, how do you dispose of it, do you have the mother sign a "release of remains". In the state of Vermont, a fetal death is 20 weeks and over OR 400 grams and over and we require an autopsy permit for this. If it is smaller than 400 grams or less than 20 weeks, it is considered a surgical specimen. Of course, the reason I am asking is our hospital was recently sued for disposing of a 200 gram fetus after following our Histology policy of disposing of surgical specimens six weeks from accession date. We are naturally "gun-shy" at this point to even discard any "products of conception". Your wisdom and guidance will be truly appreciated. Thanks, Lynne Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 From Joanholtz <@t> aol.com Tue Dec 28 12:31:43 2004 From: Joanholtz <@t> aol.com (Joanholtz@aol.com) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] RE:bor practical Message-ID: <42.5f8644a3.2f03008f@aol.com> Thanks everyone so much for all your words of wisdom and quick responses. I am glad to know that there is such a community out there willing to offer their thoughts and support (and over the holidays no less). I was taking your comments and comparing the results of my two exams. In the areas of presentation, organization and fixation. I did not have any problems. I am very much a detailed oriented person. All of my points were deducted from microtomy and special stains. When I retook the practical I was sure to note the comments from the first practical. I made sure to be as precise as before as well as taking their comments and improving on what I had submitted prior, I was very flexible. The results of the second practical were better and I had fixed my previous errors. This time categories in which I had no problem before had points deducted. I guess I'll have to come up with the right combination, or I'll have to find out where I can get it. (Gareth, you'll be the first to know) I'm curious if there are actual standard controls which our practical slides are measured up against, or is it an arbitrary process of the evaluator to determine the adequacy of the slides. I think there needs to be more transparency in the evaluation process. Thanks again for all your thoughts, Joan From Jackie.O'Connor <@t> abbott.com Tue Dec 28 12:43:47 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Fetal Remains Message-ID: In my experience, for any products of conception, it is customary and the responsibility of the 'generating' department, i.e., nursing floor, ER, OB, OR - to obtain a waiver/release or other documentation allowing the hospital to dispose of fetal remains OR surgically obtained specimens. This goes for non-OB specimens as well. I once had a motorcyclist ask for the bones from his severed toes (from a wreck) so he could make a necklace. We said no - he signed the release. I worked in a hospital where the Mom changed her mind after a couple of weeks, and wanted to have a service for her fetus. We did everything we could to find the already discarded specimen - even though she had signed the release, and we were not legally responsible to retrieve it. We did find it, and were able to return it. People need closure, sometimes. Jackie O' "Bell, Lynne" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/28/2004 12:23 PM To: cc: Subject: [Histonet] Fetal Remains I am interested how other hospital handle and/or dispose of fetal remains. In particular, I am interested in "products of conception" where there is an actual formed fetus. How long do you retain this specimen, how do you dispose of it, do you have the mother sign a "release of remains". In the state of Vermont, a fetal death is 20 weeks and over OR 400 grams and over and we require an autopsy permit for this. If it is smaller than 400 grams or less than 20 weeks, it is considered a surgical specimen. Of course, the reason I am asking is our hospital was recently sued for disposing of a 200 gram fetus after following our Histology policy of disposing of surgical specimens six weeks from accession date. We are naturally "gun-shy" at this point to even discard any "products of conception". Your wisdom and guidance will be truly appreciated. Thanks, Lynne Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GauchV <@t> mail.amc.edu Tue Dec 28 13:15:56 2004 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Blocks for Send Out Studies Message-ID: Teresa, We explain to the patient that we are required to keep the blocks but that we would be happy to send whatever slides the insititution needed to do their studies or review. We will make recut slides, unstained slides,etc. to accomodate the needs of the institution receiving the slides. It would be a VERY rare case that an original paraffin block would be sent out. Vicki >>> "Flores, Teresa" 12/28/2004 12:47:43 PM >>> Vicki, what about if a patient request that his/her blocks be sent to another institution as well. Teresa -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vicki Gauch Sent: Tuesday, December 28, 2004 9:13 AM To: malek.j@ghc.org; histonet@pathology.swmed.edu Subject: Re: [Histonet] Blocks for Send Out Studies Jack, I can only tell you that we do not release our paraffin blocks either. We offer unstained slides or 25 micron sections in a test tube,etc.- anything we can reasonably do to help them out but the blocks remain in our custody. We started this several years ago because we had issues with blocks not being returned upon completion of studies or being lost in shipping to us. There have been very few times when we have not been able to fill a request for the requesting institutions using our current system. Vicki Gauch Albany Medical Center Albany, NY >>> "Malek, Jack M" 12/27/2004 6:03:27 PM >>> Hi Everyone, Recently learned that some pathology labs send out slide sections in place of blocks for reference studies. They withhold the blocks for obvious reasons such as; to prevent losing tissue in transport, to determine which studies to exhaust the block for, preventing laser destruction of the block associated with certain molecular studies. Is this becoming the standard in practice? Happy Holidays, Jack GHC -Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BrealK <@t> Alexian.net Tue Dec 28 13:26:40 2004 From: BrealK <@t> Alexian.net (Kari Breal) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Improper tissue processing Message-ID: I have had another tissue malfunction. The last time was July 4th weekend and it happened again last weekend- Christmas weekend. The tissue came off of the processor mushy- I suspect with water still in them. The processor did not malfunction or error according to the blank error log- we have a VIP E300. When I checked the reagents in the processor- the reagents up to the last 100% were correct. The last 100% alcohol was at 87%. Which to me is what caused the raw tissue but what I can not figure out is how it got that way. This happened on Friday with minimal staff. No one changed any reagents on Friday and the tissue from Friday morning was normal. The processor was due to be rotated on Monday the 27th. My doc's are looking for reasons and answers and I am drawing a blank. I was hoping another histologist outside of this chaos might be able to think of questions or reasons that I am not currently able to. I also would like to know how everyone is treating their under processed tissue. Reprocessing by hand? Reprocessing on the VIP? We actually reprocessed using a microwave processor. The tissue turned from mush to extremely hard tissues. Unfortunately 45 of the 50 blocks were prostate bx's and there was very little tissue to begin with. Thank you for your assistance! Kari Breal From Nancy.Temple <@t> ssfhs.org Tue Dec 28 13:47:34 2004 From: Nancy.Temple <@t> ssfhs.org (Temple Nancy) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Re:Fetal remains Message-ID: <4862D47C7409844E9E52532B25609062010153FA@ontexind01.ssfhs.org> Since our hospital is a catholic hospital, all "POC" specimens under 20 weeks are retained and periodically sent to a funeral home for cremation. The families are informed of this, and releases are signed. If a family requests a private burial of a less than 20 week fetus, a gross only exam is done and the fetus released to a funeral home. Once a year, on All Saints Day, there is a service at this funeral home that all of the families that lost these babies may attend. We have a bereavement co-ordinator for OB services that handles most of the details. All POC specimens, even where there are no grossly visible fetal remains are included in the cremation. We keep a separate log of the POC specimens that tell us when specimen was received, when it was bagged, and when it was sent for cremation. N. Temple St. Francis Hospital Indianapolis, In ____________________________________________________________________________ __________________________________ The information contained in this email and any accompanying documents is intended for the sole use of the recipient to whom it is addressed, and may contain information that is privileged, confidential, and prohibited from disclosure under applicable law. If you are not the intended recipient, or authorized to receive this on behalf of the recipient, you are hereby notified that any review, use, disclosure, copying, or distribution is prohibited. If you are not the intended recipient(s), please contact the sender by e-mail and destroy all copies of the original message. Thank you. From mrsgbd2001 <@t> yahoo.com Tue Dec 28 13:49:25 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] BOR Practical Message-ID: <20041228194925.52910.qmail@web52703.mail.yahoo.com> Okay, I'm sure Joan sent in her best work, into both exams, as well as I did. I would not be complaining if I sent in mediocre work. I spent months getting properly fixed tissue and hours cutting sections. I did my GMS, AFB. PAS. Reticulin and H&E stains (only H&E done on an auto stainer) more times then I'd like to think about. I had co-workers (certified and uncertified), pathologist and supervisors review my slides before I sent them in. I am also very critical of my own work, I have had micrographs of my work published in research journals. My co-workers, who sent in their practicals at the same time, also repeated and reviewed their work. After speaking to many people, some from NSH - who actually do the grading, I think the testing process is selective. There should be a uniform format. There were seven of us using the same tissue, processors and staining techniques, yet the grades were anywhere from 700 to 393. I just want people to realize, just because we didn't pass our practical it doesn't mean we are laid-back and don't care about our work. Okay, I'm done. Thanks, Gareth Davis Histo tech Nashville, Tn --------------------------------- Do you Yahoo!? Send a seasonal email greeting and help others. Do good. From kbradshaw <@t> lcpath.com Tue Dec 28 13:42:50 2004 From: kbradshaw <@t> lcpath.com (Kari Bradshaw) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Improper tissue processing In-Reply-To: Message-ID: It's ironic how your problem has occurred over two holiday weekends. Do you run a different program over the holiday weekends? When we have had to reprocess under processed tissue we melt the blocks down, and reversed process on the VIP cleaning cycle. Then we reprocess on a short run excluding formalin and starting with 70% alcohol. This protocol was published in the Histologic Technical Bulletin by Sakura in Nov 2001. Unfortunately we have become pretty good at it. Kari Bradshaw, HT(ASCP) Laboratory Manager Lower Columbia Pathologists 1217 14th Ave Longview, WA 98632 (360)425-5620 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kari Breal Sent: Tuesday, December 28, 2004 11:27 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Improper tissue processing I have had another tissue malfunction. The last time was July 4th weekend and it happened again last weekend- Christmas weekend. The tissue came off of the processor mushy- I suspect with water still in them. The processor did not malfunction or error according to the blank error log- we have a VIP E300. When I checked the reagents in the processor- the reagents up to the last 100% were correct. The last 100% alcohol was at 87%. Which to me is what caused the raw tissue but what I can not figure out is how it got that way. This happened on Friday with minimal staff. No one changed any reagents on Friday and the tissue from Friday morning was normal. The processor was due to be rotated on Monday the 27th. My doc's are looking for reasons and answers and I am drawing a blank. I was hoping another histologist outside of this chaos might be able to think of questions or reasons that I am not currently able to. I also would like to know how everyone is treating their under processed tissue. Reprocessing by hand? Reprocessing on the VIP? We actually reprocessed using a microwave processor. The tissue turned from mush to extremely hard tissues. Unfortunately 45 of the 50 blocks were prostate bx's and there was very little tissue to begin with. Thank you for your assistance! Kari Breal _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Linresearch <@t> aol.com Tue Dec 28 13:51:40 2004 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] (no subject) Message-ID: <74.49fc0e1b.2f03134c@aol.com> Hello, Are there any antibodies available that will differentiate fragmented collagen from total collagen? Lin From Julie.Sanders <@t> med.va.gov Tue Dec 28 13:19:45 2004 From: Julie.Sanders <@t> med.va.gov (Julie.Sanders@med.va.gov) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] RE: Histonet Digest, Vol 13, Issue 33 Message-ID: <457381D92B01BD44B21CF37CC02EBDFD02927550@vhacinexc2.v10.med.va.gov> Ole, The procedure for the Sevier-Munger can be found on page 257 of Theory and Practice of Histotechnology by Sheehan, Hrapchak, 1980. If you don't have this book, I'd be happy to fax the procedure to you or send it email. Julie Julie Sanders, BA, HT(ASCP) Supervisor, Anatomic Pathology VAMC, Cincinnati, Ohio ------------------------------ Message: 11 Date: Tue, 28 Dec 2004 16:29:34 +0100 From: "Ole" Subject: [Histonet] Sevier Munger Silver stain To: Message-ID: <000e01c4ecf2$0a2c9070$0100000a@brukere> Content-Type: text/plain; charset="iso-8859-1" Hi I need a protocol/receipe of the Sevier Munger silver stain. I have not found it in any of my histotech books nor had any luck searching the web. I work with PCR, immunohistochemistry and in situs and dont know much about special stains, or where to find them, if their not in our "special stain" books. It is to be used on Formaldehyd fixed paraffin embedded tissue. Best regards Ole ------------------------------ Message: 12 Date: Tue, 28 Dec 2004 07:36:46 -0800 (PST) From: Gareth Davis Subject: Re: [Histonet] please advise: bor practical To: lpwenk@sbcglobal.net, Histonet Message-ID: <20041228153646.66909.qmail@web52705.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii thanks for information on the HT Practical. I did realize that there were other criteria involved in the grading, but most of my comments were on staining. Anyway, I thought about taking the HTL, but it seems most people I know only have an HT and are doing almost as well in the field. Thanks again, Gareth Davis lpwenk@sbcglobal.net wrote: Just wondering - Did you take the HTL exam and the rest of the people take the HT exam? Since you have a BS? The HTL exam IS harder than the HT exam - larger tissues sections, trickier stains, often needing to cut thinner tissues, tissues that are more difficult to obtain, etc. I train both HT and HTL students, and can attest that the HTL exam is harder to do than the HT exam. Peggy A. Wenk, HTL(ASCP)SLS Program Director, Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Gareth Davis" To: ; "Histonet" Sent: Monday, December 27, 2004 5:48 PM Subject: Re: [Histonet] please advise: bor practical > Hey Joan, > I'm in the exact same boat as you. Our lab does all > work manual except the H&E stain. There were seven of > us, from my lab, taking the test this fall. Out of the > seven I was the only one to fail. (I am also sending > my slides in for review.) I don't see how I could > have failed - based on the staining of three of my H&E > slides - when I used the same H&E machine my > co-workers used. Not that a degree gives a person > more "talent", but I am the only one of the seven with > a B.S., and I wonder if that they scored me on a > higher scale - which would not seem fair. I was told > that the evaluation process was anonymous, by a member > of NSH. > I would also be interested in what happens with the > reevaluation process. > > Good luck to you. > Gareth > > --- Joanholtz@aol.com wrote: > > > The hospital in which I work uses manual processes > > for all of our cutting and > > special staining needs. The only automation we use > > is the H&E. I would > > think this is the preferred way to learn hands on > > and was quite proud to learn > > this way. I was assured that after passing the > > written portion of the exam, > > (which I did on my first attempt) that the practical > > should be no problem at all. > > I have submitted my practical twice; this time I am > > eligible to submit my > > slides for a reevaluation along with more money. > > Before each submission my > > slides were reviewed by at least two of our > > pathologists. If 800 is the optimal > > score I do not think it is reasonable to assume that > > my hospitals lab puts out > > or accepts anything below a 300. I would truly > > appreciate any and all help or > > insights as I try and appeal my scores by Jan. 10th > > 2005. > > > > Am I at a disadvantage without automation? How > > should I select slides to be > > reevaluated? Is it possible to get a worse score? > > Am I wasting my time? Is > > the evaluation process anonymous, do they know your > > background? > > > > This is truly a very frustrating situation with a BS > > in Biology and three > > years histology experience. I have invested a lot of > > time, energy and money into > > getting where I am now and this arbitrary process is > > preventing me from going > > any further. > > > > Thank you for any and all help, > > Joan > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - You care about security. So do we. > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 13 Date: Tue, 28 Dec 2004 07:41:30 -0800 From: "Robyn Vazquez" Subject: Re: [Histonet] Sevier Munger Silver stain To: histonet@lists.utsouthwestern.edu, osteffe@online.no Message-ID: Content-Type: text/plain; charset=us-ascii Ole, Do you have fax no.? I have recipe for Sevier-Munger Method for neural tissues Robyn Ohsu >>> "Ole" 12/28/2004 7:29:34 AM >>> Hi I need a protocol/receipe of the Sevier Munger silver stain. I have not found it in any of my histotech books nor had any luck searching the web. I work with PCR, immunohistochemistry and in situs and dont know much about special stains, or where to find them, if their not in our "special stain" books. It is to be used on Formaldehyd fixed paraffin embedded tissue. Best regards Ole _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Mon, 27 Dec 2004 10:44:38 -0800 From: "FIGUEIREDO,MARXA L " Subject: [Histonet] help To: histonet@lists.utsouthwestern.edu Message-ID: <1104173078.41d05816c1428@mail.ucla.edu> Content-Type: text/plain; charset=ISO-8859-1 Hi HIstonetters, Has anyone done imunohistochemisty with anti-GFP in paraffin tissues recently? I have the anti-GFP monoclonal from Clontech (JL-8) and was wondering if anyone has tried that one in FFPE tissues. If so, what was the dilution and antigen retrieval method? THanks Marxa Figueiredo UCLA Dentistry ------------------------------ Message: 15 Date: Tue, 28 Dec 2004 09:54:54 -0600 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] Sevier Munger Silver stain To: "Ole" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" It's in the AFIP manual. The reference is; Sevier, A. C., and Munger, B. L.: J. Neuropath. Exp. Neurol. 24:130-135, 1965. Sorry but I don't have time to type the whole thing. 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 Ole Sent: Tuesday, December 28, 2004 9:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sevier Munger Silver stain Hi I need a protocol/receipe of the Sevier Munger silver stain. I have not found it in any of my histotech books nor had any luck searching the web. I work with PCR, immunohistochemistry and in situs and dont know much about special stains, or where to find them, if their not in our "special stain" books. It is to be used on Formaldehyd fixed paraffin embedded tissue. Best regards Ole _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Tue, 28 Dec 2004 11:44:28 -0500 From: MTitford@aol.com Subject: [Histonet] On passing BOR exam To: histonet@lists.utsouthwestern.edu Message-ID: <3F49D0EC.73796666.00762DB1@aol.com> Content-Type: text/plain; charset=iso-8859-1 Joan Holtz asks about passing the BOR exam. I was shocked a few years ago when I read the ASCP newsletter that gave statistics for the registry exams. Many histotechs failed the HT and HTL practicals. Now, not talking about Joans laboratory or anyone else's in particular, I suspect many applicants bang out their exam slides like they do their daily work slides and send them off without critically examining them first. In our laboratory we receive slides for consultation which are sub-standard. Obviously those laboratories think they are acceptable. With HT and HTL practical exam slides you should produce the best d*** slides you can! You have plenty of time. Scrounge for the best fixed tissues. Submit tissues of right size allowing for shrinkage. Process correctly. Cut correctly. Produce best stained slides you can. In histology, it is easy to produce adequate slides, but hard to produce perfect slides.The slides should be of right thickness, no scores, folds, precipitates, background staining etc. The staining should be perfect! Remember, if you submit a poorly stained slide, you are telling the examiner thats you best work. In my laboratory when preparing for practical exams, I expect histotechs to make multiple blocks, and multiple stained slides from those blocks. We sit down together several times and look closely at those slides. There is much going back and re-submitting tissue. We have only had one histotech fail the practical in 25 years (And that tech did not want me to see those slides!) Mike Titford USA Pathology Mobile AL USA ------------------------------ Message: 17 Date: Tue, 28 Dec 2004 11:59:02 -0500 From: MTitford@aol.com Subject: [Histonet] Argyrophil stains. To: histonet@lists.utsouthwestern.edu Message-ID: <49AF28FD.5B6B2BA3.00762DB1@aol.com> Content-Type: text/plain; charset=iso-8859-1 Ole, somewhere overseas I suspect, asks about the Sevier Munger stain. The reference is: Sevier, A.C., Munger, B.L., A silver method for paraffin sections of neural tissue. J. Neuropath Exp Neurol. 24: 130 - 135 1965 However, in the USA (and is it not interesting how labs use procedures written by their own countrymen?)many labs use the Churukian method which demonstrates a wider variety of argyrophil tumors. Churukian, J.,Schenk, E.A. A modification of Pascual's argyrophil method. J. Histotechnol 3: 102 - 102 1979. I read somewhere, long ago, that there is no one method which will demonstrate ALL argyophil tumors, but the Chrukian method demonstrates the widest variety. Mike Titford USA Pathology Mobile AL USA ------------------------------ Message: 18 Date: Tue, 28 Dec 2004 12:32:48 -0500 From: "Favara, Cynthia (NIH/NIAID)" Subject: RE: [Histonet] help To: "'FIGUEIREDO,MARXA L '" , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain I have been doing anti GFP in mouse FFPE tissue. I am presently using Molecular Probes Rabbit polyclonal 2mg/ml at a dilution of 1:500 using a biotinylated secondary from Vector @1:250 dil on the Ventana Nexus. I use Citrate pH 6.0 in a pressure cooker temp 120 for 30 seconds. I will be evaluating monoclonal from Zymed and BD Biosciences when I get the time. Sorry I have no experience with Clontech. 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: FIGUEIREDO,MARXA L [mailto:mlfiguei@ucla.edu] Sent: Monday, December 27, 2004 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] help Hi HIstonetters, Has anyone done imunohistochemisty with anti-GFP in paraffin tissues recently? I have the anti-GFP monoclonal from Clontech (JL-8) and was wondering if anyone has tried that one in FFPE tissues. If so, what was the dilution and antigen retrieval method? THanks Marxa Figueiredo UCLA Dentistry _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Tue, 28 Dec 2004 11:49:32 -0600 From: "Charles.Embrey" Subject: RE: [Histonet] On passing BOR exam To: , Message-ID: Content-Type: text/plain; charset="us-ascii" Thank-you! Thank-you! Thank-you! I am shocked by the laid-back attitude that many people today take in approaching the practical exam. They don't seem to even read the grading criteria required to produce the slides capable of passing. Each slide sent in to be graded should first be graded by the technician against these grading guidelines. There should really be no surprises. If you don't know enough about histology to grade your own slides you really shouldn't be certified as a histotech. As far as automation goes I think all practical exam slides should be produced WITHOUT automation. I don't ever want to hire a histotech that can't do a GMS by hand. Sorry if this seems harsh but as a lab manager I want to see the BOR stamp of approval mean something. Quit wringing your hands; take a look at the list of errors provided by the grader and produce slides that avoid those mistakes. Charles Embrey, Pathologists' Assistant, HT(ASCP) Histology Manager Carle Clinic, Urbana Illinois -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of MTitford@aol.com Sent: Tuesday, December 28, 2004 10:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] On passing BOR exam Joan Holtz asks about passing the BOR exam. I was shocked a few years ago when I read the ASCP newsletter that gave statistics for the registry exams. Many histotechs failed the HT and HTL practicals. Now, not talking about Joans laboratory or anyone else's in particular, I suspect many applicants bang out their exam slides like they do their daily work slides and send them off without critically examining them first. In our laboratory we receive slides for consultation which are sub-standard. Obviously those laboratories think they are acceptable. With HT and HTL practical exam slides you should produce the best d*** slides you can! You have plenty of time. Scrounge for the best fixed tissues. Submit tissues of right size allowing for shrinkage. Process correctly. Cut correctly. Produce best stained slides you can. In histology, it is easy to produce adequate slides, but hard to produce perfect slides.The slides should be of right thickness, no scores, folds, precipitates, background staining etc. The staining should be perfect! Remember, if you submit a poorly stained slide, you are telling the examiner thats you best work. In my laboratory when preparing for practical exams, I expect histotechs to make multiple blocks, and multiple stained slides from those blocks. We sit down together several times and look closely at those slides. There is much going back and re-submitting tissue. We have only had one histotech fail the practical in 25 years (And that tech did not want me to see those slides!) Mike Titford USA Pathology Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 13, Issue 33 **************************************** From RossS <@t> BaylorHealth.edu Tue Dec 28 14:05:21 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Fetal Remains Message-ID: <17A8DA5A05167C4EA20BFA750167736B0BB6BA@BHDAEXCH13.bhcs.pvt> At my former hospital in Maryland, we required a disposal form separate for fetuses before we would accession the specimen. The only exception was if the family was planning a funeral. Now occasionally we would get a case where the family said they were having a funeral home pick it up and they didn't. Those would usually sit for at least a year before a Pathologist would give permission to dispose of it. Here under Texas law all specimens must be disposed of as Pathological waste or by a licensed funeral home. We have people ask for things all the time, but I just quote the Texas Law (25 Tex. Admin. Code 1.132 Section 1.133). Currently we do not have a separate disposal form for fetuses. We are creating one because the hospital is supposed to start funding funeral home pick up of fetuses and scattering of ashes on an area of ground in a local cemetery. The new form will give 3 choices for fetal disposal: their own funeral home, the hospital funded cremation and scattering of ashes, or disposal as Pathological Waste (although the chaplains are going to find a nicer term for it). Your legal or risk management department should be able to at least assist you in creating a form that lessen the chance of another lawsuit. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie M O'Connor Sent: Tuesday, December 28, 2004 12:44 PM To: Bell, Lynne Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Fetal Remains In my experience, for any products of conception, it is customary and the responsibility of the 'generating' department, i.e., nursing floor, ER, OB, OR - to obtain a waiver/release or other documentation allowing the hospital to dispose of fetal remains OR surgically obtained specimens. This goes for non-OB specimens as well. I once had a motorcyclist ask for the bones from his severed toes (from a wreck) so he could make a necklace. We said no - he signed the release. I worked in a hospital where the Mom changed her mind after a couple of weeks, and wanted to have a service for her fetus. We did everything we could to find the already discarded specimen - even though she had signed the release, and we were not legally responsible to retrieve it. We did find it, and were able to return it. People need closure, sometimes. Jackie O' "Bell, Lynne" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/28/2004 12:23 PM To: cc: Subject: [Histonet] Fetal Remains I am interested how other hospital handle and/or dispose of fetal remains. In particular, I am interested in "products of conception" where there is an actual formed fetus. How long do you retain this specimen, how do you dispose of it, do you have the mother sign a "release of remains". In the state of Vermont, a fetal death is 20 weeks and over OR 400 grams and over and we require an autopsy permit for this. If it is smaller than 400 grams or less than 20 weeks, it is considered a surgical specimen. Of course, the reason I am asking is our hospital was recently sued for disposing of a 200 gram fetus after following our Histology policy of disposing of surgical specimens six weeks from accession date. We are naturally "gun-shy" at this point to even discard any "products of conception". Your wisdom and guidance will be truly appreciated. Thanks, Lynne Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From RBARNHART <@t> summithealth.org Tue Dec 28 15:06:11 2004 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Xylene resistant labels Message-ID: We are looking into using blank xylene resistant labels that we would print the information on. Is there anyone out there doing this? What printer and labels do you use? Thanks Becky From stewart_georgia <@t> sbcglobal.net Tue Dec 28 15:23:26 2004 From: stewart_georgia <@t> sbcglobal.net (Georgia Stewart) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Xylene resistant labels In-Reply-To: Message-ID: <20041228212326.46177.qmail@web81408.mail.yahoo.com> Becky At the lab I used to work at, we obtained our preprinted xylene resistant labels from Surgipath. Used these labels preprinted with the Pap accession number and name of the laboratory for the Pap slides. Surgipath's prices for plain or preprinted labels were much lower than what we could obtain from a local source. Georgia Stewart, BS, HT, HTL stewart_georgia@sbcglobal.net Rebecca Barnhart wrote: We are looking into using blank xylene resistant labels that we would print the information on. Is there anyone out there doing this? What printer and labels do you use? Thanks Becky _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Tue Dec 28 15:38:40 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Xylene resistant labels Message-ID: <83AACDB0810528418AA106F9AE9B7F7E5077BA@sjhaexc02.sjha.org> We get our heat/xylene resistant labels from Deryk Upton Printers Plus Inc Toll Free Phone 800-667-1512 Toll Free Fax 877-434-0884 We use Zebra printers. These work well for us. Good luck, Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rebecca Barnhart Sent: Tuesday, December 28, 2004 4:06 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Xylene resistant labels We are looking into using blank xylene resistant labels that we would print the information on. Is there anyone out there doing this? What printer and labels do you use? Thanks Becky _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From RCHIOVETTI <@t> aol.com Tue Dec 28 16:34:27 2004 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Histo Survey for Nevada: Last Call! Message-ID: <15a.46e6172c.2f033973@aol.com> Histonetters, I need to close the Histo survey for the state of Nevada by mid-January. If there are any practicing histotechnologists in the state who haven't responded but who would like to give input, please contact me off-list for details. Thanks and greetings from (mostly) sunny Arizona! Bob Chiovetti P.S. Here's the original post: ****************************************************** If you are a practicing histotechnologist in the state of Nevada and if you can spare about 10-15 minutes to participate in a survey, I would appreciate hearing from you. This isn't a salary survey; it's geared more towards the general "health and welfare" of the profession and the labs in the state (size of labs, number of docs on staff, case load, instrumentation, usage, service and maintenance issues and so on). I need input from *all* histo labs, whether they're hospital labs, derm/Mohs clinics, veterinary diagnostics labs, reference labs, academic/research labs, etc. You can easily complete the survey at your computer, then simply save the document and return it to me as an attachment to an e-mail message. I also have a hard copy version that I can send to you via snail-mail if you prefer. Please contact me off-list if you'd like to participate or if you have any questions. Thanks in advance. Happy Holidays to everyone! Cheers, Robert (Bob) Chiovetti, Ph.D. Tucson, AZ From dsnider <@t> shrinenet.org Wed Dec 29 07:09:55 2004 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Glass knife Makers Message-ID: My lab is getting collecting information on glass knife makers, in anticipation of purchasing one. Can anyone out there give some input on particular brands, since this is totally out of my realm. I am most interested in ease of use, durability, etc. I am having a really difficult time trying to collect information on these items. Thanks in advance.... Deanna Snider HT ASCP Research Lab Shriners Hospital for Children Cincinnati, OH CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From DonnaWillis <@t> texashealth.org Wed Dec 29 08:28:08 2004 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] SNOMED International Message-ID: Happy New Year Everyone, Does anyone know where to purchase a SNOMED International coding manual. We need to validate a new computer system and currently not using the "International" version. Thanks, Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From Sue.Kapoor <@t> uhsi.org Wed Dec 29 10:03:02 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] looking for slide dryer Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F5682@khmcexch.uhsi.org> Hi all, I'm looking to purchase a slide dryer, I would prefer the type that you open and set a rack of slides into, not the type that you lay the slides on. Demo units, rebuilts will be fine, the only catch is it MUST be under $500, can anyone help me? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From Terry.Marshall <@t> rothgen.nhs.uk Wed Dec 29 10:27:40 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] looking for slide dryer Message-ID: I have often wondered if anyone has rigged a perforated box to one of those elephant trunk women's hair driers to produce a slide dryer. Come to think of it, men probably use them to, they become such a bunch of Nancy boys. 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: Kapoor, Sue [mailto:Sue.Kapoor@uhsi.org] Sent: 29 December 2004 16:03 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] looking for slide dryer Hi all, I'm looking to purchase a slide dryer, I would prefer the type that you open and set a rack of slides into, not the type that you lay the slides on. Demo units, rebuilts will be fine, the only catch is it MUST be under $500, can anyone help me? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RCHIOVETTI <@t> aol.com Wed Dec 29 10:56:49 2004 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Glass knife Makers Message-ID: In a message dated 12/29/2004 6:14:42 AM US Mountain Standard Time, dsnider@shrinenet.org writes: My lab is getting collecting information on glass knife makers, in anticipation of purchasing one. Can anyone out there give some input on particular brands, since this is totally out of my realm. I am most interested in ease of use, durability, etc. I am having a really difficult time trying to collect information on these items. Thanks in advance.... Hi Deanna, Glass knifemakers are available for the two most commonly used types of knives: triangular knives and ralph (long) knives. The kind of knifemaker will depend on the application. If you only want to do histology at the light level, you could use either a triangular knifemaker or a ralph knifemaker. If your interest is in taking the specimens to the electron microscope you'll need to use a triangular knifemaker. You will see the differences when you visit the websites below. The two biggest manufacturers of triangular glass knifemakers are Leica and RMC Products (now part of Boeckeler Instruments). SPI Supplies also has an economical triangular knifemaker. Ralph knifemakers are available from a few specialty suppliers. Take a look at the following websites. You'll find contact info on the sites. For Ralph knifemakers: Ted Pella http://www.tedpella.com/histo_html/8702.htm For Ralph and triangular knifemakers: EBS http://www.ebsstore.com/control/category/~category_id=D2/~pcategory=D For triangular knifemakers: Leica, RMC and SPI http://www.em-preparation.com/ (This is the Leica site...you'll need to navigate as follows: Products -> Biological Specimens -> Room Temperature Techniques -> Leica EM KMR2) http://www.rmcproducts.com/glass-knife.html http://www.2spi.com/catalog/knives/glass.shtml There are a few other suppliers, but this should get you started. Good luck! Bob Chiovetti From gcallis <@t> montana.edu Wed Dec 29 11:08:19 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Re: Glass knife Makers In-Reply-To: References: Message-ID: <6.0.0.22.1.20041229095358.01b43f50@gemini.msu.montana.edu> The one we have is still made by Energy Beam Sciences and formerly the old Sorvall knifemaker design. It makes triagular knives with various thickness glass, and is extremely easy to use. I don't recall the price, but repair service is very reliable from EBS. We sent off an old breaker and had it refurbished for very little money - it was a joy to be able to do this. Go to their website or contact Cindy Gaspari at this email address. CGaspari@ebsciences.com. She is a nice lady ,very helpful and can give you pricing, etc. At 06:09 AM 12/29/2004, you wrote: >My lab is getting collecting information on glass knife makers, in >anticipation of purchasing one. Can anyone out there give some input on >particular brands, since this is totally out of my realm. I am most >interested in ease of use, durability, etc. I am having a really difficult >time trying to collect information on these items. Thanks in advance.... > >Deanna Snider HT ASCP >Research Lab >Shriners Hospital for Children >Cincinnati, OH >CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may >contain confidential and privileged information for the use of the >designated recipients. If you are not the intended recipient, (or >authorized to receive for the recipient) you are hereby notified that you >have received this communication in error and that any review, disclosure, >dissemination, distribution or copying of it or its contents is prohibited. >If you have received this communication in error, please destroy all copies >of this communication and any attachments and contact the sender by reply >e-mail or telephone (813) 281-0300. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From dsnider <@t> shrinenet.org Wed Dec 29 12:25:30 2004 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Knife maker Message-ID: We currently have an old LKB, which nobody can make a decent knife with. I will be cutting plastic embedded specimens. I will need the triangular knife. Deanna CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From Bauer.Karen <@t> mayo.edu Wed Dec 29 12:40:09 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Fetal Remains Message-ID: Hi Lynne, We save all POC's that have a fetus no matter what age or gms. We put it in a "Save" drawer and then they are buried every spring in a mass burial that we have with another hospital in town. Anything that is over the 20 wks or 400 gms needs to have an autopsy or disposal form signed by the parents if they do not want the fetus back for a family burial. The ones close to full term are kept in the morgue. The other hospital takes care of all the arrangements and contacts me as to when the funeral home will be coming to pick up the specimens. Any family who wishes to be notified of the date and time of the burial is called by our Resolve Through Sharing group. All tissues/specimens that are requested by a patient needs to be picked up by a licensed funeral director. We do not want to be responsible for what these patients are doing with these specimens. If the funeral director gives the "specimen" to the patient, that is their choice and we are not liable for anything. I hope this helps a little, Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bell, Lynne Sent: Tuesday, December 28, 2004 12:23 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fetal Remains I am interested how other hospital handle and/or dispose of fetal remains. In particular, I am interested in "products of conception" where there is an actual formed fetus. How long do you retain this specimen, how do you dispose of it, do you have the mother sign a "release of remains". In the state of Vermont, a fetal death is 20 weeks and over OR 400 grams and over and we require an autopsy permit for this. If it is smaller than 400 grams or less than 20 weeks, it is considered a surgical specimen. Of course, the reason I am asking is our hospital was recently sued for disposing of a 200 gram fetus after following our Histology policy of disposing of surgical specimens six weeks from accession date. We are naturally "gun-shy" at this point to even discard any "products of conception". Your wisdom and guidance will be truly appreciated. Thanks, Lynne Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From mcauliff <@t> umdnj.edu Wed Dec 29 15:53:08 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Knife maker In-Reply-To: References: Message-ID: <41D32744.7020002@umdnj.edu> The LKB knifemaker is a great tool. In my experience poor quality knives are due to the machine being out of adjustment. The scoring pressure could be too low (or too high), you might need a new scoring wheel or the clamping head may be applying insufficient pressure. Also, the score may not be in the proper position for a good break.. Find the instructions (a fold-up laminated sheet that fits in a slot under the machine), get a glass strip and take an hour to figure out what the problem is. I would be surprised if the instrument was beyond repair. Geoff Snider, Deanna wrote: >We currently have an old LKB, which nobody can make a decent knife with. I >will be cutting plastic embedded specimens. I will need the triangular >knife. > >Deanna >CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may >contain confidential and privileged information for the use of the >designated recipients. If you are not the intended recipient, (or >authorized to receive for the recipient) you are hereby notified that you >have received this communication in error and that any review, disclosure, >dissemination, distribution or copying of it or its contents is prohibited. >If you have received this communication in error, please destroy all copies >of this communication and any attachments and contact the sender by reply >e-mail or telephone (813) 281-0300. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From lu_ze <@t> sbcglobal.net Wed Dec 29 17:58:41 2004 From: lu_ze <@t> sbcglobal.net (Ze Lu) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] need advice on Leica TP 1050 tissue processor Message-ID: <00d801c4ee02$5255b370$1302a8c0@optimum2> Histonet friends, We are planning to buy a used Leica TP 1050 tissue processor. Not decide yet. I haven't used this type tissue processor. We want to have inexpensive, reliable, flexible program control and easy to use tissue processor. Does anyone has some advice on this type? We appreciate your input and helps. By the way, happy new year to everyone. Ze Lu, Ph.D. Optimum Therapeutics, LLC From TJJ <@t> Stowers-Institute.org Wed Dec 29 15:20:45 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Re: Need assistance with in situ protocol!!! (LONG) Message-ID: Sorry it's taken me so long to reply, I've been unsubscribed for a couple weeks! Two things I think would help. Proteinase K digestion is optimal when you can limit the time in the enzyme by altering concentrations. We use 5 ug/ml for 7 minutes at 37 degrees C on our paraffin sections (7 microns thick), and that works fine for us. This might help keep your tissue on the slides a bit better. Additionally, we use the secondary antibody (sheep anti-dig) at 1:2000 dilution overnight at room temperature. I think your secondary antibody concentration is too strong and that might be causing some of your problems. Also do a blast search on your sequence and see if perhaps both the sense and anti-sense are recognizing something in your sample (specific binding). As for commercial RNA probes, I haven't found any. You might try finding a sequence to GapDH or actin for positive control. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From customerservice <@t> easterscientific.com Wed Dec 29 15:32:47 2004 From: customerservice <@t> easterscientific.com (customerservice@easterscientific.com) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Happy New Year Message-ID: ESICO wants to take this opportunity to wish everyone a safe, happy and prosperous new year! Remember to contact ESICO for all of your histology instrument repair and maintenance needs, including tissue processors, embedders, microtomes, cryostats, stainers, coverslippers and microscopes. ESICO services the following brands: Leica, Leitz, Ames/Miles, A.O., Reichert, Jung, Olympus, Nikon and Zeiss. Please call if you don't see your brand listed -- chances are we can help! We travel nationwide and we offer competitive pricing. Warm Regards, ESICO Easter Scientific Instrument Co. 2212 East 12th Street, Suite 224 Davenport, IA 52803 Tel. 888-322-2391 (toll free) Fax 563-326-6047 From mari.ann.mailhiot <@t> leica-microsystems.com Wed Dec 29 15:48:07 2004 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] need advice on Leica TP 1050 tissue processor Message-ID: Ze Lu You can give me a call here at Leica and I can provide a schedule to optimize your use of reagents for processing on the Leica TP1050. I also have several processing program that may fit your needs. Best Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com "Ze Lu" Sent by: To: histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] need advice on Leica TP 1050 tissue processor 12/29/2004 05:58 PM Histonet friends, We are planning to buy a used Leica TP 1050 tissue processor. Not decide yet. I haven't used this type tissue processor. We want to have inexpensive, reliable, flexible program control and easy to use tissue processor. Does anyone has some advice on this type? We appreciate your input and helps. By the way, happy new year to everyone. Ze Lu, Ph.D. Optimum Therapeutics, LLC _______________________________________________ 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 histo007 <@t> hotmail.com Wed Dec 29 15:55:58 2004 From: histo007 <@t> hotmail.com (Jim Ball) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Needle biopsies Message-ID: I am a tech with 25+ years of experience and have been bitten by about every snake in the garden of eden (Histology), and I guess that is one of the main reasons I will error on the side of caution at every turn. I really try to be as conservative as possible with tissue when trimming into a needle biopsy, as soon as I have a full face on properly enbedded needles(usually not more than 20 microns or less I start taking slides). The sections are 3microns and may produce as many as 5 to 10 sections suitable for mounting. This acounts for max 30 more micrones into the block. It is at this point I would like to preserve the remainder of the tissue until it is reviewed by a pathologist. I refer to my madness as scouting (a procedure if used by General Custer would have saved alot of lives), but as we all know there are some patologist that will declare we did not trim enough if what they are looking for is 100 micrones into the block. While I have been reseaching a procedure that will keep everyone happy I ran across an article that state there was a study done to determine if histologists were trimming away microcalcifications in needle biopsies, and according to the high lights of the article (one they wanted me to purchase to add insult to injury) it was determined that after x-raying the histology shavings from trimmed breast biopsies the culprit once again was the histologist. Go figure. At the present time I am on a public computer and some one needs to use it, but before I leave please foward any ideas you may have on this subject via this server or directly to my e-mail address listed with this posting From emry <@t> u.washington.edu Wed Dec 29 19:56:13 2004 From: emry <@t> u.washington.edu (Trisha Emry) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Methyl salicylate/glycerin Message-ID: Hi, We have some specimens that have been clearing in Methyl salicylate. We would like to transfer them to storage in glycerin. Are there anti-fungal additives that are needed for that purpose? The methyl salicylate is considered a hazard by our environmental health and safety people. Are there tricks to cleaning up things that have been used in it. Or a way to make it safer to handle? Thanks and Happy 2005! Trisha U. of Washington From jkiernan <@t> uwo.ca Wed Dec 29 21:02:35 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Methyl salicylate/glycerin References: Message-ID: <41D36FCB.5F3D5FD0@uwo.ca> Methyl salicylate is not miscible with glycerol. Both liquids are miscible with ethanol, so pass the specimens through a few changes of 100% alcohol in transit from meth sal to glycerol. If your environmental health and safety people think meth sal is hazardous, they should be able to give you expert advice about avoiding the supposed risks. Meth sal is also called oil of wintergreen; its MSDS sheet (http://sargentwelch.com/pdf/msds/sch94614.pdf) does not indicate any notable dangers. The smell, though quite pleasant, can be annoying if you're around the stuff for a long time. Benzyl benzoate is an almost odourless solvent that is optically similar to meth sal. (The slightly higher refractive index of benz benz makes it slightly superior to meth sal for examining whole specimens; both solvents give much better transparency than glycerol.) John Kiernan London, Canada. ____________________________________________ Trisha Emry wrote: > > Hi, > > We have some specimens that have been clearing in Methyl salicylate. We > would like to transfer them to storage in glycerin. Are there anti-fungal > additives that are needed for that purpose? > > The methyl salicylate is considered a hazard by our environmental health and > safety people. Are there tricks to cleaning up things that have been used > in it. Or a way to make it safer to handle? > > Thanks and Happy 2005! > > Trisha > U. of Washington > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Thu Dec 30 05:22:26 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Needle biopsies Message-ID: My personal opinion is: It is always easier to go back and get deeper levels on the block later, than to lose an irreplaceable biopsy by trimming too much into it. Let the pathologists bitch all they want. 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 Jim Ball Sent: Wednesday, December 29, 2004 3:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Needle biopsies I am a tech with 25+ years of experience and have been bitten by about every snake in the garden of eden (Histology), and I guess that is one of the main reasons I will error on the side of caution at every turn. I really try to be as conservative as possible with tissue when trimming into a needle biopsy, as soon as I have a full face on properly enbedded needles(usually not more than 20 microns or less I start taking slides). The sections are 3microns and may produce as many as 5 to 10 sections suitable for mounting. This acounts for max 30 more micrones into the block. It is at this point I would like to preserve the remainder of the tissue until it is reviewed by a pathologist. I refer to my madness as scouting (a procedure if used by General Custer would have saved alot of lives), but as we all know there are some patologist that will declare we did not trim enough if what they are looking for is 100 micrones into the block. While I have been reseaching a procedure that will keep everyone happy I ran across an article that state there was a study done to determine if histologists were trimming away microcalcifications in needle biopsies, and according to the high lights of the article (one they wanted me to purchase to add insult to injury) it was determined that after x-raying the histology shavings from trimmed breast biopsies the culprit once again was the histologist. Go figure. At the present time I am on a public computer and some one needs to use it, but before I leave please foward any ideas you may have on this subject via this server or directly to my e-mail address listed with this posting _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Thu Dec 30 06:01:01 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Needle biopsies Message-ID: <83AACDB0810528418AA106F9AE9B7F7E5077D8@sjhaexc02.sjha.org> Perhaps you can cut a specified amount such as 50 microns between levels and then xray the block if you do not find the calcifications in the first cut sections. Then you can be sure there are some before you cut through the block. Maybe your pathologists would be agreeable to that? Good luck! Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of GUTIERREZ, JUAN Sent: Thursday, December 30, 2004 6:22 AM To: Jim Ball; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Needle biopsies My personal opinion is: It is always easier to go back and get deeper levels on the block later, than to lose an irreplaceable biopsy by trimming too much into it. Let the pathologists bitch all they want. 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 Jim Ball Sent: Wednesday, December 29, 2004 3:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Needle biopsies I am a tech with 25+ years of experience and have been bitten by about every snake in the garden of eden (Histology), and I guess that is one of the main reasons I will error on the side of caution at every turn. I really try to be as conservative as possible with tissue when trimming into a needle biopsy, as soon as I have a full face on properly enbedded needles(usually not more than 20 microns or less I start taking slides). The sections are 3microns and may produce as many as 5 to 10 sections suitable for mounting. This acounts for max 30 more micrones into the block. It is at this point I would like to preserve the remainder of the tissue until it is reviewed by a pathologist. I refer to my madness as scouting (a procedure if used by General Custer would have saved alot of lives), but as we all know there are some patologist that will declare we did not trim enough if what they are looking for is 100 micrones into the block. While I have been reseaching a procedure that will keep everyone happy I ran across an article that state there was a study done to determine if histologists were trimming away microcalcifications in needle biopsies, and according to the high lights of the article (one they wanted me to purchase to add insult to injury) it was determined that after x-raying the histology shavings from trimmed breast biopsies the culprit once again was the histologist. Go figure. At the present time I am on a public computer and some one needs to use it, but before I leave please foward any ideas you may have on this subject via this server or directly to my e-mail address listed with this posting _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From JNocito <@t> Pathreflab.com Thu Dec 30 07:00:14 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Needle biopsies In-Reply-To: Message-ID: Jim, I always refer to what my 8th grade shop teacher told us. It is always easier to shave off a little more wood, than it is trying to glue it back on. I have taken this advice to the histo lab. Once, when I was a rookie many moons ago, I had to fish through paraffin shavings to locate a renal biopsy that I cut away. Since that time, I always cut on the conservative side and explain to the pathologists that it always easier to go back into the block than fish through paraffin shavings. I do get static sometimes, but after they cool down, they realize it was a good descion. As always, the opinions of the author do not reflect the opinions of his employers and their lawyers. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jim Ball Sent: Wednesday, December 29, 2004 3:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Needle biopsies I am a tech with 25+ years of experience and have been bitten by about every snake in the garden of eden (Histology), and I guess that is one of the main reasons I will error on the side of caution at every turn. I really try to be as conservative as possible with tissue when trimming into a needle biopsy, as soon as I have a full face on properly enbedded needles(usually not more than 20 microns or less I start taking slides). The sections are 3microns and may produce as many as 5 to 10 sections suitable for mounting. This acounts for max 30 more micrones into the block. It is at this point I would like to preserve the remainder of the tissue until it is reviewed by a pathologist. I refer to my madness as scouting (a procedure if used by General Custer would have saved alot of lives), but as we all know there are some patologist that will declare we did not trim enough if what they are looking for is 100 micrones into the block. While I have been reseaching a procedure that will keep everyone happy I ran across an article that state there was a study done to determine if histologists were trimming away microcalcifications in needle biopsies, and according to the high lights of the article (one they wanted me to purchase to add insult to injury) it was determined that after x-raying the histology shavings from trimmed breast biopsies the culprit once again was the histologist. Go figure. At the present time I am on a public computer and some one needs to use it, but before I leave please foward any ideas you may have on this subject via this server or directly to my e-mail address listed with this posting _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From flemons <@t> bhset.org Thu Dec 30 08:04:07 2004 From: flemons <@t> bhset.org (Fran Lemons) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] BOR practical Message-ID: Can anyone tell me of a certainty whether or not explanations are given on the written notification of a failed practical? It was my understanding that it is included with your results, but I have a candidate who says there isn't any explanation on hers. Thanks in advance Fran Walker From ryaskovich <@t> dir.nidcr.nih.gov Thu Dec 30 08:17:05 2004 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR)) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Needle biopsies Message-ID: <8F3AB322628548428A992EFB0E80F5D3A2F227@nihexchange8.nih.gov> Joe I agree 100%! Ruth Yaskovich National Institutes of Health > ---------- > From: Joe Nocito > Sent: Thursday, December 30, 2004 7:00 AM > To: Jim Ball; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Needle biopsies > > Jim, > I always refer to what my 8th grade shop teacher told us. It is always > easier to shave off a little more wood, than it is trying to glue it back > on. I have taken this advice to the histo lab. > Once, when I was a rookie many moons ago, I had to fish through > paraffin > shavings to locate a renal biopsy that I cut away. Since that time, I > always > cut on the conservative side and explain to the pathologists that it > always > easier to go back into the block than fish through paraffin shavings. I do > get static sometimes, but after they cool down, they realize it was a good > descion. > > As always, the opinions of the author do not reflect the opinions of his > employers and their lawyers. > > > Joe Nocito, BS, HT(ASCP) QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jim Ball > Sent: Wednesday, December 29, 2004 3:56 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Needle biopsies > > > I am a tech with 25+ years of experience and have been bitten by about > every > snake in the garden of eden (Histology), and I guess that is one of the > main > reasons I will error on the side of caution at every turn. I really try to > be as conservative as possible with tissue when trimming into a needle > biopsy, as soon as I have a full face on properly enbedded needles(usually > not more than 20 microns or less I start taking slides). The sections are > 3microns and may produce as many as 5 to 10 sections suitable for > mounting. > This acounts for max 30 more micrones into the block. It is at this point > I > would like to preserve the remainder of the tissue until it is reviewed by > a > pathologist. I refer to my madness as scouting (a procedure if used by > General Custer would have saved alot of lives), but as we all know there > are some patologist that will declare we did not trim enough if what they > are looking for is 100 micrones into the block. > While I have been reseaching a procedure that will keep everyone happy > I > ran across an article that state there was a study done to determine if > histologists were trimming away microcalcifications in needle biopsies, > and > according to the high lights of the article (one they wanted me to > purchase > to add insult to injury) it was determined that after x-raying the > histology > shavings from trimmed breast biopsies the culprit once again was the > histologist. Go figure. > At the present time I am on a public computer and some one needs to use > it, but before I leave please foward any ideas you may have on this > subject > via this server or directly to my e-mail address listed with this posting > > > > _______________________________________________ > Histonet mailing list > Histonet@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 Bauer.Karen <@t> mayo.edu Thu Dec 30 09:35:16 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Needle biopsies Message-ID: Hi Jim, This is the way we treat all of our needle biopsies: As soon as we face into the block and see some exposed tissue, we start taking our sections. We cut 8 slides (charged slides), two levels on each slide. For example; face in a little, take the first section, face in a little more, take the next section, and so on until 8 slides have two levels on each of them. The rule of thumb here is to start with a little tissue and end up with a little tissue so the pathologists know that all of the biopsy has been sampled. We then stain slides 1, 3, 5, and 7 with H&E and save 2, 4, 6, and 8. These saved slides are for possible specials or IP's. One pathologist just has us stain the rest with H&E if he doesn't need any further stains. This seems to keep the pathologists happy, we have had no complaints. All of the saved slides are kept for a month or so, just in case, otherwise they are thrown away. Hope this helps you out a bit and good luck, Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jim Ball Sent: Wednesday, December 29, 2004 3:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Needle biopsies I am a tech with 25+ years of experience and have been bitten by about every snake in the garden of eden (Histology), and I guess that is one of the main reasons I will error on the side of caution at every turn. I really try to be as conservative as possible with tissue when trimming into a needle biopsy, as soon as I have a full face on properly enbedded needles(usually not more than 20 microns or less I start taking slides). The sections are 3microns and may produce as many as 5 to 10 sections suitable for mounting. This acounts for max 30 more micrones into the block. It is at this point I would like to preserve the remainder of the tissue until it is reviewed by a pathologist. I refer to my madness as scouting (a procedure if used by General Custer would have saved alot of lives), but as we all know there are some patologist that will declare we did not trim enough if what they are looking for is 100 micrones into the block. While I have been reseaching a procedure that will keep everyone happy I ran across an article that state there was a study done to determine if histologists were trimming away microcalcifications in needle biopsies, and according to the high lights of the article (one they wanted me to purchase to add insult to injury) it was determined that after x-raying the histology shavings from trimmed breast biopsies the culprit once again was the histologist. Go figure. At the present time I am on a public computer and some one needs to use it, but before I leave please foward any ideas you may have on this subject via this server or directly to my e-mail address listed with this posting _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Thu Dec 30 10:44:52 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Methyl salicylate Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3C6B@exchsrv01.barrynet.barry.edu> If the health and safety people had their way, oxygen dihydride would be classified as hazardous. Methyl salicylate, a.k.a. oil of wintergreen, is used to flavor some mint candies. In quantity, it is toxic, but the oral LD 50 for humans is around 1 ounce (30 ml). Tabasco sauce is equally toxic. Spilled on the skin, methyl salicylate create a mild irritation and warming of the skin (erythema) that some people find pleasant. Swimmers used to rub themselves down with methyl salicylate as part of the warm-up for competition. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Trisha Emry Sent: Wednesday, December 29, 2004 8:56 PM To: histonet@lists.utsouthwestern.edu Cc: emry@u.washington.edu Subject: [Histonet] Methyl salicylate/glycerin Hi, We have some specimens that have been clearing in Methyl salicylate. We would like to transfer them to storage in glycerin. Are there anti-fungal additives that are needed for that purpose? The methyl salicylate is considered a hazard by our environmental health and safety people. Are there tricks to cleaning up things that have been used in it. Or a way to make it safer to handle? Thanks and Happy 2005! Trisha U. of Washington _______________________________________________ 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 jdmd77 <@t> hotmail.com Thu Dec 30 10:46:03 2004 From: jdmd77 <@t> hotmail.com (Julia Dahl) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] Needle biopsies In-Reply-To: Message-ID: Wanted to chime in from a pathologist perspective. I've always found it useful to think of each piece of tissue as a patient, and to let that guide my requests and management. I depend on histologists for the very quality of my work and the patients that I am rendering diagnoses on - and I try to remember that daily and treat our relationship as such. I need you guys!!!! As I've become more seasoned, the whole risk management/lawsuit avoidance mentality has influenced my practice - as it seems to have influenced some of the other pathologists you are dealing with. The study referred to in this thread appears to blame the histologist for microcalcifications that were cut through and potentially discarded. Unless there is a alteration in the procedure and the pathologist and histologist working together - what's the point of blame and shame? Sounds pretty crappy. It's true that any legal action for the cut away microcalcifications will rest squarely with the pathologist. Rather than blaming the histologist (misery loves company) it seems like a good time to be working WITH the histologist in the best interest of the patient. I don't enjoy the position that the decreasing amounts of tissue and clinician demands for diagnosis nonetheless leave all of us in. But our role is to serve our patients, to facilitate communication between the clinician and the patient and to accomplish this - we (pathologists and histologists) must be a unified team. How often do you really experience this? Do you have time or the opportunity to be at the microscope with your pathologists? Probably not. The reality in most practices is: If you surface cut the block - you'll be told to "go deeper" - if you throw away a level, you'll be told "what the hell were you thinking?" If you give the pathologist 8 slides, we're likely to claim "slide innundation!!!!" TOO MUCH WORK!! You're damed if you do and damned if you don't. Is there a winning situation? It seems very prudent to surface cut the block initially, but if diagnostic features aren't present histologically, there is the risk that the block will be recut on a different microtome, causing unnecessary loss of tissue. If you use this methodology, there MUST be a safeguard in place such that the same histologist and microtome are used to cut the deepers. And the pathologists have to be "on board" with the policy (particularly if the person cutting recuts until 3:00 p.m. won't be allowed to cut the needle core biopsy unless the pathologist signs a waiver of the policy - I'm sure many of you are getting red-faced pathologist mental images). [Yeah, we can be an "interesting" bunch, hm?] The Mayo method of cutting all the tissue on the same microtome up front is one solution. Something to keep in mind from a risk management standpoint, though, is that if there is any legal action - some lawyer is going to see slides labelled 1, 3, 5 and 7 and say "where are 2, 4, 6 and 8." Then someone is going to write a check to the plaintiff. At UWashington (where I trained and did a breast/immuno fellowship) - all the needle core biopsies were trimmed close to the tissue, to provide ribbons on each slide (usually 6 - 8 on biopsies less than 1.5 cm), and we reviewed two slides. Recuts were kind of a disaster, since UW didn't have a policy in place when I was there (1999) to ensure the same histologist and microtome. I suppose that besides offering up some risk management perspective and a bit of humor - the only other perspective that I can share is to use these situations to BUILD relationship with your team of histologists (communicate, communicate, communicate) and with your pathologists. We have more in common (we work for the sake of the patient) than we have differences. Julia Dahl MD >From: "Bauer, Karen" >To: "Jim Ball" , >Subject: RE: [Histonet] Needle biopsies >Date: Thu, 30 Dec 2004 09:35:16 -0600 >Hi Jim, > >This is the way we treat all of our needle biopsies: > >As soon as we face into the block and see some exposed tissue, we start >taking our sections. We cut 8 slides (charged slides), two levels on >each slide. For example; face in a little, take the first section, face >in a little more, take the next section, and so on until 8 slides have >two levels on each of them. The rule of thumb here is to start with a >little tissue and end up with a little tissue so the pathologists know >that all of the biopsy has been sampled. We then stain slides 1, 3, 5, >and 7 with H&E and save 2, 4, 6, and 8. These saved slides are for >possible specials or IP's. One pathologist just has us stain the rest >with H&E if he doesn't need any further stains. This seems to keep the >pathologists happy, we have had no complaints. All of the saved slides >are kept for a month or so, just in case, otherwise they are thrown >away. > >Hope this helps you out a bit and good luck, > >Karen Bauer HT(ASCP) >Histology Supervisor >Luther Hospital >Eau Claire, WI > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jim Ball >Sent: Wednesday, December 29, 2004 3:56 PM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Needle biopsies > >I am a tech with 25+ years of experience and have been bitten by about >every snake in the garden of eden (Histology), and I guess that is one >of the main reasons I will error on the side of caution at every turn. I >really try to be as conservative as possible with tissue when trimming >into a needle biopsy, as soon as I have a full face on properly enbedded >needles(usually not more than 20 microns or less I start taking slides). >The sections are 3microns and may produce as many as 5 to 10 sections >suitable for mounting. >This acounts for max 30 more micrones into the block. It is at this >point I would like to preserve the remainder of the tissue until it is >reviewed by a pathologist. I refer to my madness as scouting (a >procedure if used by General Custer would have saved alot of lives), but >as we all know there are some patologist that will declare we did not >trim enough if what they are looking for is 100 micrones into the block. > While I have been reseaching a procedure that will keep everyone >happy I ran across an article that state there was a study done to >determine if histologists were trimming away microcalcifications in >needle biopsies, and according to the high lights of the article (one >they wanted me to purchase to add insult to injury) it was determined >that after x-raying the histology shavings from trimmed breast biopsies >the culprit once again was the histologist. Go figure. > At the present time I am on a public computer and some one needs to >use it, but before I leave please foward any ideas you may have on this >subject via this server or directly to my e-mail address listed with >this posting > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Thu Dec 30 11:04:25 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:26 2005 Subject: [Histonet] BOR practical References: Message-ID: <002301c4ee91$9f4e6480$fff2ff44@domainnotset.invalid> When she received her notification of not passing, she should have also received a listing of all the tissues and stains, with series of numbers after each one. These numbers the "errors" listed on pages 6, 7 and 8 of the practical booklet she received months ago, that listed what stains and tissues she was to send in. For example, if the tissue results were: Fallopian Tube, H&E - 203, 206, 412 This would mean - on an H&E of the fallopian tube: 203 = nuclear stain not crisp 206 = cytoplasmic differentiation slightly deficient 412 = knife lines If she did not save her practical booklet, she can download the booklet from the ASCP Board of Registry webpage: http://www.ascp.org/bor/getcertified/index.asp Click on either HT or HTL Then click on Practical Exam Instructions - this booklet is for the 2005 practical tissue list, but "errors" are the same as 2004. Peggy A. Wenk, HTL(ASCP)SLS Program Director, Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Fran Lemons" To: Sent: Thursday, December 30, 2004 9:04 AM Subject: [Histonet] BOR practical Can anyone tell me of a certainty whether or not explanations are given on the written notification of a failed practical? It was my understanding that it is included with your results, but I have a candidate who says there isn't any explanation on hers. Thanks in advance Fran Walker _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Dorothy.L.Webb <@t> HealthPartners.Com Thu Dec 30 11:05:31 2004 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] processing help Message-ID: We are adding an additional processor to our lab and I would like to know if anyone uses an alternative fixative on any of their processors for "fatty" specimens, lymph nodes, etc. and if so, could you please EMail me what you use and your processing schedule? I would like to try to get the most optimal fixationa nd processing out of our large, problematic specimens now that we have the capacity to do this. Any helpful hints would be greatly appreciated! Thank you all ahead of time and "Happy New Year" to all of my "brother and sister" histotechs! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From kbroomal <@t> NEMOURS.ORG Thu Dec 30 12:01:57 2004 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] BOR practical Message-ID: <9BCAC308B27CAD4196D8AC9ADF037AAA110A78E3@wlmmsx01.nemours.org> I don't think those score breakdowns were sent out yet. I passed both sections, but I just received my overall score for the computer section & practical. I do remember reading that we should get a breakdown. I hope that maybe that comes later? Kristen Broomall -----Original Message----- From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] Sent: Thursday, December 30, 2004 12:04 PM To: Fran Lemons; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] BOR practical When she received her notification of not passing, she should have also received a listing of all the tissues and stains, with series of numbers after each one. These numbers the "errors" listed on pages 6, 7 and 8 of the practical booklet she received months ago, that listed what stains and tissues she was to send in. For example, if the tissue results were: Fallopian Tube, H&E - 203, 206, 412 This would mean - on an H&E of the fallopian tube: 203 = nuclear stain not crisp 206 = cytoplasmic differentiation slightly deficient 412 = knife lines If she did not save her practical booklet, she can download the booklet from the ASCP Board of Registry webpage: http://www.ascp.org/bor/getcertified/index.asp Click on either HT or HTL Then click on Practical Exam Instructions - this booklet is for the 2005 practical tissue list, but "errors" are the same as 2004. Peggy A. Wenk, HTL(ASCP)SLS Program Director, Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Fran Lemons" To: Sent: Thursday, December 30, 2004 9:04 AM Subject: [Histonet] BOR practical Can anyone tell me of a certainty whether or not explanations are given on the written notification of a failed practical? It was my understanding that it is included with your results, but I have a candidate who says there isn't any explanation on hers. Thanks in advance Fran Walker _______________________________________________ Histonet mailing list Histonet@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 HACKERLAB <@t> aol.com Thu Dec 30 12:33:49 2004 From: HACKERLAB <@t> aol.com (HACKERLAB@aol.com) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Happy New Year Message-ID: <1ad.2e184cea.2f05a40d@aol.com> Best Wishes to All for a Happy New Year! >From all of us at Hacker Instruments & Industries Inc Winnsboro, South Carolina From Marion.Hiles <@t> nbt.nhs.uk Wed Dec 29 09:08:36 2004 From: Marion.Hiles <@t> nbt.nhs.uk (Marion Hiles) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Improper tissue processing Message-ID: <2EE924DF60902943AC6E2EF35155451F19CD4C@nbfexch03.north-bristol.nhs> Thats very odd. How do you know the last 100% alcohol was actually 87%? We have a VIP E300.How often do you replace the reagents and do you bump up the reagents, so that the last of, say, the 3 100% alcohols is the freshest? Bob Quilty Neuropath Frenchay Hospital Bristol UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kari Breal Sent: 28 December 2004 19:27 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Improper tissue processing I have had another tissue malfunction. The last time was July 4th weekend and it happened again last weekend- Christmas weekend. The tissue came off of the processor mushy- I suspect with water still in them. The processor did not malfunction or error according to the blank error log- we have a VIP E300. When I checked the reagents in the processor- the reagents up to the last 100% were correct. The last 100% alcohol was at 87%. Which to me is what caused the raw tissue but what I can not figure out is how it got that way. This happened on Friday with minimal staff. No one changed any reagents on Friday and the tissue from Friday morning was normal. The processor was due to be rotated on Monday the 27th. My doc's are looking for reasons and answers and I am drawing a blank. I was hoping another histologist outside of this chaos might be able to think of questions or reasons that I am not currently able to. I also would like to know how everyone is treating their under processed tissue. Reprocessing by hand? Reprocessing on the VIP? We actually reprocessed using a microwave processor. The tissue turned from mush to extremely hard tissues. Unfortunately 45 of the 50 blocks were prostate bx's and there was very little tissue to begin with. Thank you for your assistance! Kari Breal _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: The information in this message is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorised. If you are not the intended recipient, any disclosure, copying, or distribution of the message, or any action or omission taken by you in reliance on it, is prohibited and may be unlawful. Please immediately contact the sender if you have received this message in error. Thank you. From starostm <@t> ors.od.nih.gov Wed Dec 29 10:13:45 2004 From: starostm <@t> ors.od.nih.gov (Starost, Matthew (NIH/OD/ORS)) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Immunogold Message-ID: <8B08CB90735BE243B6B7ED7CA0074C14128829E5@nihexchange6.nih.gov> I am looking for protocols for immunogold staining for transmission EM purposes. I would like a protocol for DNA and for histone. We are evaluating mouse sperm. I have not been able to find an antibody in the US. Any and all help would be greatly appreciated. Matthew F. Starost, DVM, PhD, Diplomate ACVP Veterinary Pathologist National Institutes of Health Division of Veterinary Resources Building 28A, Room 115 9000 Rockville Pike Bethesda, Maryland 20892 Phone: 301-451-2176 Fax: 301-402-1068 starostm@mail.nih.gov From lpwenk <@t> sbcglobal.net Thu Dec 30 13:28:46 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] BOR practical References: <9BCAC308B27CAD4196D8AC9ADF037AAA110A78E3@wlmmsx01.nemours.org> Message-ID: <001001c4eea5$c8d60420$fff2ff44@domainnotset.invalid> If a candidate passes the practical, they receive their score, but NO breakdown. If a candidate does NOT pass the practical, they receive their score AND the breakdown, to give guidance on what needs improving, for when they retake the practical. Congratulations on passing both parts, by the way! Peggy A. Wenk, HTL(ASCP)SLS Program Director, Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48037 ----- Original Message ----- From: "Kristen Broomall" To: Sent: Thursday, December 30, 2004 1:01 PM Subject: RE: [Histonet] BOR practical > I don't think those score breakdowns were sent out yet. I passed both > sections, but I just received my overall score for the computer section & > practical. I do remember reading that we should get a breakdown. I hope that > maybe that comes later? > > Kristen Broomall > > -----Original Message----- > From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] > Sent: Thursday, December 30, 2004 12:04 PM > To: Fran Lemons; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] BOR practical > > > When she received her notification of not passing, she should have also > received a listing of all the tissues and stains, with series of numbers > after each one. These numbers the "errors" listed on pages 6, 7 and 8 of the > practical booklet she received months ago, that listed what stains and > tissues she was to send in. > > For example, if the tissue results were: > Fallopian Tube, H&E - 203, 206, 412 > This would mean - on an H&E of the fallopian tube: > 203 = nuclear stain not crisp > 206 = cytoplasmic differentiation slightly deficient > 412 = knife lines > > If she did not save her practical booklet, she can download the booklet from > the ASCP Board of Registry webpage: > > http://www.ascp.org/bor/getcertified/index.asp > Click on either HT or HTL > Then click on Practical Exam Instructions - this booklet is for the 2005 > practical tissue list, but "errors" are the same as 2004. > > Peggy A. Wenk, HTL(ASCP)SLS > Program Director, Schools of Histotechnology > William Beaumont Hospital > Royal Oak, MI 48073 > > > > ----- Original Message ----- > From: "Fran Lemons" > To: > Sent: Thursday, December 30, 2004 9:04 AM > Subject: [Histonet] BOR practical > > > Can anyone tell me of a certainty whether or not explanations are given on > the written notification of a failed practical? It was my understanding > that it is included with your results, but I have a candidate who says there > isn't any explanation on hers. > Thanks in advance > Fran Walker > > > _______________________________________________ > Histonet mailing list > Histonet@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 Michael.Rice <@t> holy-cross.com Thu Dec 30 13:30:49 2004 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Improper tissue processing Message-ID: <3BC92F29BE821745AB15E04C98EE028D6936EA@HCH2KMAIL.holy-cross.com> It sounds as though whoever bumped the reagents placed a lower proof alcohol in the 100% slot. I have worked with vip for many years and have not seen a rotary valve problem that would account for this mike rice holy cross hospital ft lauderdale, fl ( where it is warm and sunny) -----Original Message----- From: Marion Hiles [mailto:Marion.Hiles@nbt.nhs.uk] Sent: Wednesday, December 29, 2004 10:09 AM To: 'Kari Breal' Cc: Histonet (E-mail) Subject: RE: [Histonet] Improper tissue processing Thats very odd. How do you know the last 100% alcohol was actually 87%? We have a VIP E300.How often do you replace the reagents and do you bump up the reagents, so that the last of, say, the 3 100% alcohols is the freshest? Bob Quilty Neuropath Frenchay Hospital Bristol UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kari Breal Sent: 28 December 2004 19:27 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Improper tissue processing I have had another tissue malfunction. The last time was July 4th weekend and it happened again last weekend- Christmas weekend. The tissue came off of the processor mushy- I suspect with water still in them. The processor did not malfunction or error according to the blank error log- we have a VIP E300. When I checked the reagents in the processor- the reagents up to the last 100% were correct. The last 100% alcohol was at 87%. Which to me is what caused the raw tissue but what I can not figure out is how it got that way. This happened on Friday with minimal staff. No one changed any reagents on Friday and the tissue from Friday morning was normal. The processor was due to be rotated on Monday the 27th. My doc's are looking for reasons and answers and I am drawing a blank. I was hoping another histologist outside of this chaos might be able to think of questions or reasons that I am not currently able to. I also would like to know how everyone is treating their under processed tissue. Reprocessing by hand? Reprocessing on the VIP? We actually reprocessed using a microwave processor. The tissue turned from mush to extremely hard tissues. Unfortunately 45 of the 50 blocks were prostate bx's and there was very little tissue to begin with. Thank you for your assistance! Kari Breal _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: The information in this message is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorised. If you are not the intended recipient, any disclosure, copying, or distribution of the message, or any action or omission taken by you in reliance on it, is prohibited and may be unlawful. Please immediately contact the sender if you have received this message in error. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- From kbroomal <@t> NEMOURS.ORG Thu Dec 30 14:16:46 2004 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] BOR practical Message-ID: <9BCAC308B27CAD4196D8AC9ADF037AAA110A78E9@wlmmsx01.nemours.org> Thanks! Well, that's a bummer! I wanted to know how I did on each part. Especially that computer test!! ;) Kristen Broomall -----Original Message----- From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] Sent: Thursday, December 30, 2004 2:29 PM To: Kristen Broomall; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] BOR practical If a candidate passes the practical, they receive their score, but NO breakdown. If a candidate does NOT pass the practical, they receive their score AND the breakdown, to give guidance on what needs improving, for when they retake the practical. Congratulations on passing both parts, by the way! Peggy A. Wenk, HTL(ASCP)SLS Program Director, Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48037 ----- Original Message ----- From: "Kristen Broomall" To: Sent: Thursday, December 30, 2004 1:01 PM Subject: RE: [Histonet] BOR practical > I don't think those score breakdowns were sent out yet. I passed both > sections, but I just received my overall score for the computer section & > practical. I do remember reading that we should get a breakdown. I hope that > maybe that comes later? > > Kristen Broomall > > -----Original Message----- > From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] > Sent: Thursday, December 30, 2004 12:04 PM > To: Fran Lemons; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] BOR practical > > > When she received her notification of not passing, she should have also > received a listing of all the tissues and stains, with series of numbers > after each one. These numbers the "errors" listed on pages 6, 7 and 8 of the > practical booklet she received months ago, that listed what stains and > tissues she was to send in. > > For example, if the tissue results were: > Fallopian Tube, H&E - 203, 206, 412 > This would mean - on an H&E of the fallopian tube: > 203 = nuclear stain not crisp > 206 = cytoplasmic differentiation slightly deficient > 412 = knife lines > > If she did not save her practical booklet, she can download the booklet from > the ASCP Board of Registry webpage: > > http://www.ascp.org/bor/getcertified/index.asp > Click on either HT or HTL > Then click on Practical Exam Instructions - this booklet is for the 2005 > practical tissue list, but "errors" are the same as 2004. > > Peggy A. Wenk, HTL(ASCP)SLS > Program Director, Schools of Histotechnology > William Beaumont Hospital > Royal Oak, MI 48073 > > > > ----- Original Message ----- > From: "Fran Lemons" > To: > Sent: Thursday, December 30, 2004 9:04 AM > Subject: [Histonet] BOR practical > > > Can anyone tell me of a certainty whether or not explanations are given on > the written notification of a failed practical? It was my understanding > that it is included with your results, but I have a candidate who says there > isn't any explanation on hers. > Thanks in advance > Fran Walker > > > _______________________________________________ > Histonet mailing list > Histonet@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 RCHIOVETTI <@t> aol.com Thu Dec 30 14:19:09 2004 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Immunogold Message-ID: <7e.6006880b.2f05bcbd@aol.com> In a message dated 12/30/2004 12:22:26 PM US Mountain Standard Time, starostm@ors.od.nih.gov writes: I am looking for protocols for immunogold staining for transmission EM purposes. I would like a protocol for DNA and for histone. We are evaluating mouse sperm. I have not been able to find an antibody in the US. Matthew, Setting aside the histone labeling issues for a moment, would the osmium ammine reaction be suitable for the DNA work? It provides a quite intense, electron-dense reaction product for TEM, and it avoids all of the problems with trying to find an anti-DNA antibody, coupling the AB to some form of gold tracer, etc. There have been a few studies that indicate the labeling of DNA with osmium ammine is preferable to using immunogold, and osmium ammine-B is readily available from Polysciences (Product is Osmium ammine-B, Cat. No. 48016-91-7). Just a thought. Best, Bob Chiovetti From waltersk <@t> mail.medicine.uiowa.edu Thu Dec 30 14:58:56 2004 From: waltersk <@t> mail.medicine.uiowa.edu (Walters, Katherine S) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] help from derm people (skin-heads?) Message-ID: Hello experts- I've been using skin for a control tissue in some of my antibody studies and my PI is distressed that the corneum layer is disconnected in spots from the rest of the tissue. (BTW-I am paraffin processing the skin) I've always thought that this was "normal" for dermal tissues. Is there any way to prevent the separation? TIA, Kathy From AnthonyH <@t> chw.edu.au Thu Dec 30 15:34:11 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Needle biopsies Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E151@simba.kids> I wholeheartedly agree Regards Tony Henwood JP, 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: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Friday, 31 December 2004 12:00 AM To: Jim Ball; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Needle biopsies Jim, I always refer to what my 8th grade shop teacher told us. It is always easier to shave off a little more wood, than it is trying to glue it back on. I have taken this advice to the histo lab. Once, when I was a rookie many moons ago, I had to fish through paraffin shavings to locate a renal biopsy that I cut away. Since that time, I always cut on the conservative side and explain to the pathologists that it always easier to go back into the block than fish through paraffin shavings. I do get static sometimes, but after they cool down, they realize it was a good descion. As always, the opinions of the author do not reflect the opinions of his employers and their lawyers. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jim Ball Sent: Wednesday, December 29, 2004 3:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Needle biopsies I am a tech with 25+ years of experience and have been bitten by about every snake in the garden of eden (Histology), and I guess that is one of the main reasons I will error on the side of caution at every turn. I really try to be as conservative as possible with tissue when trimming into a needle biopsy, as soon as I have a full face on properly enbedded needles(usually not more than 20 microns or less I start taking slides). The sections are 3microns and may produce as many as 5 to 10 sections suitable for mounting. This acounts for max 30 more micrones into the block. It is at this point I would like to preserve the remainder of the tissue until it is reviewed by a pathologist. I refer to my madness as scouting (a procedure if used by General Custer would have saved alot of lives), but as we all know there are some patologist that will declare we did not trim enough if what they are looking for is 100 micrones into the block. While I have been reseaching a procedure that will keep everyone happy I ran across an article that state there was a study done to determine if histologists were trimming away microcalcifications in needle biopsies, and according to the high lights of the article (one they wanted me to purchase to add insult to injury) it was determined that after x-raying the histology shavings from trimmed breast biopsies the culprit once again was the histologist. Go figure. At the present time I am on a public computer and some one needs to use it, but before I leave please foward any ideas you may have on this subject via this server or directly to my e-mail address listed with this posting _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From Alde.Gavino <@t> UTSouthwestern.edu Thu Dec 30 17:00:12 2004 From: Alde.Gavino <@t> UTSouthwestern.edu (Alde Gavino) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Mouse Keratins Message-ID: Hello all, I am starting a project on alopecia areata and will be working on mouse hair follicles. Before studying the expression pattern of the molecule we are interested in, I first would like to work on a marker that is known to be expressed on hair follicle epithelium. In this light, I want to ask if anyone knows whether or not monoclonal antibodies against mouse keratin 5 or 14 are commercially availbale for the purpose of immunohistochemistry/immunofluorescence staining. I would appreciate very much your help! Thank you so much! Sincerely, Carlo Gavino From carl.hobbs <@t> kcl.ac.uk Fri Dec 31 09:54:57 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] re need advice on Leica TP 1050 tissue processor Message-ID: I have had one for 5yrs and have just bought another one. Very easy to use. Mine has the perspex skirting around it as part of the fume control system. I can have it in the open lab because of this. I have two programmes; 22 and 44 hrs. This processes everything very well. From st18 chick embryos, decalcified mouse mandibles to complete adult rat brain.I use gradations of Methylated spirit to dehydrate and xylene to clear. No vacuum. A good, reliable ( for me)simple machine. --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.822 / Virus Database: 560 - Release Date: 22/12/2004 From Rcartun <@t> harthosp.org Fri Dec 31 10:00:22 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Coverslipping problems Message-ID: Dear Histonet colleagues: We have a Sakura Tissue-Tek SCA coverslipper that we use for our immunoperoxidase stained slides. In the past it worked great. Now, we constantly have bubbles and dried-out areas (I think you refer to it as "cornflaking") on the slides. Apparently, we do not have these same problems with our H&E-stained slides. Any suggestions? Best wishes for a happy, healthy, and peaceful New Year to everyone! 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 ohenry <@t> dfw.net Fri Dec 31 16:27:20 2004 From: ohenry <@t> dfw.net (Susan Owens) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Needle biopsies Message-ID: <002201c4ef87$f76dad00$71dd3040@Nationwide.net> Jim, I've been reading the threads sent on 'needle biopsies' and I'm surprised no one has suggested you x-ray the blocks BEFORE you start cutting. For many years now, when we have breast tissue( needle location) all the blocks(6 or 20,etc) are set in numerical order on a plastic tray and x-rayed. The radiologist reads the film and checks the blocks believed to contain calcium. When cutting, we level the blocks marked by the radiologist. Seems like the same thing may be possible for your ' needle biopsies'. Having an x-ray film of your material/block, before you start cutting, may answer allot of questions. We take the blocks down to x-ray and the tech takes a picture. Then they call us to come and get it AFTER the doctor has read the film. SO, we now have blocks AND film for our pathologists. All together it takes about an hour to get our blocks and film. If you have access to x-ray try it and let us know. Susan ---------------------------------------------------------------------------- ---- Message: 10 Date: Fri, 31 Dec 2004 08:34:11 +1100 From: Tony Henwood Subject: RE: [Histonet] Needle biopsies To: "'Joe Nocito'" , Jim Ball , histonet@lists.utsouthwestern.edu Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E151@simba.kids> Content-Type: text/plain I wholeheartedly agree Regards Tony Henwood JP, 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: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Friday, 31 December 2004 12:00 AM To: Jim Ball; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Needle biopsies Jim, I always refer to what my 8th grade shop teacher told us. It is always easier to shave off a little more wood, than it is trying to glue it back on. I have taken this advice to the histo lab. Once, when I was a rookie many moons ago, I had to fish through paraffin shavings to locate a renal biopsy that I cut away. Since that time, I always cut on the conservative side and explain to the pathologists that it always easier to go back into the block than fish through paraffin shavings. I do get static sometimes, but after they cool down, they realize it was a good descion. As always, the opinions of the author do not reflect the opinions of his employers and their lawyers. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jim Ball Sent: Wednesday, December 29, 2004 3:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Needle biopsies I am a tech with 25+ years of experience and have been bitten by about every snake in the garden of eden (Histology), and I guess that is one of the main reasons I will error on the side of caution at every turn. I really try to be as conservative as possible with tissue when trimming into a needle biopsy, as soon as I have a full face on properly enbedded needles(usually not more than 20 microns or less I start taking slides). The sections are 3microns and may produce as many as 5 to 10 sections suitable for mounting. This acounts for max 30 more micrones into the block. It is at this point I would like to preserve the remainder of the tissue until it is reviewed by a pathologist. I refer to my madness as scouting (a procedure if used by General Custer would have saved alot of lives), but as we all know there are some patologist that will declare we did not trim enough if what they are looking for is 100 micrones into the block. While I have been reseaching a procedure that will keep everyone happy I ran across an article that state there was a study done to determine if histologists were trimming away microcalcifications in needle biopsies, and according to the high lights of the article (one they wanted me to purchase to add insult to injury) it was determined that after x-raying the histology shavings from trimmed breast biopsies the culprit once again was the histologist. Go figure. At the present time I am on a public computer and some one needs to use it, but before I leave please foward any ideas you may have on this subject via this server or directly to my e-mail address listed with this posting From vmckaughan <@t> ccmh.org Thu Dec 30 12:09:22 2004 From: vmckaughan <@t> ccmh.org (Vicki Mckaughan) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] using VIP processor for processing small biopsies Message-ID: <000001c4f1b5$bd5d4f00$69010180@MEGAFIRE.COM> I am looking for a shortened processing schedule using the VIP 3000 tissue processor to process small biopsy specimens, especially GI biopsies. We are currently trying to adjust our run, but are primarily having trouble with adenomatous polyps being fragmented and shattered. Does anyone have any suggestions? Please e-mail me at vlm_23@yahoo.com thanks, Vicki McKaughan Histology Supervisor Camden Clark Memorial Hospital