From Marjorie.Lehman <@t> unilever.com Mon Nov 1 05:54:08 2004 From: Marjorie.Lehman <@t> unilever.com (marjorie lehman) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Offtopic - Microscope for kids Message-ID: Please post to H-net too. I have an 8 yo g-daughter and she wants one too. -----Original Message----- From: Kim Merriam [SMTP:kmerriam2003@yahoo.com] Sent: Friday, October 29, 2004 8:18 PM To: Histonet Subject: [Histonet] Offtopic - Microscope for kids Hello all, My 8 year old son wants a microscope for Christmas. I have been searching on the internet for a decent one that does not cost an arm and a leg. It seems like there are lots of cheap ones in the $12-30 range and then some nice ones that are in the $100-200 range. Does anyone know where I could get one that costs around $50-70 that is not a piece of junk? I would like 2 or 3 objectives. Of course, the slides need not be included! Thanks in advance, Kim Merriam Novartis Cambridge, MA Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hymclab <@t> hyhc.com Mon Nov 1 07:52:58 2004 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Antigen Retrieval w/ B&D Steamer..... Message-ID: Here is what we do-- I got this off the histonet about 1 1/2 years ago and it works wonderful. Turn on rice steamer and put in coplin jar or jars of retrieval solution while running down slides (this is sufficient time to get the solution to the right temp., approx. 90 degrees C) Place solutions into retrival solution(being sure to place 7 slides in each coplin jar even if you need to add blank slides for fillers) and set rice steamer for 20 minutes. After 20 minutes take out coplin jars and let cool for 10 minutes. Proceed with staining. This is a great procedure and provides very consistent staining. Hope this helps, Dawn Schneider, HT(ASCP) Howard Young Medical Center Woodruff, WI -----Original Message----- From: Brandon Stokes [mailto:cuttingedgehistology@yahoo.com] Sent: Friday, October 29, 2004 6:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Antigen Retrieval w/ B&D Steamer..... Hi fellow Histonetters........ I have a quick question for anyone out there that knows anything about using a Black&Decker Steamer for Antigen Retrieval. How long do you pre-heat the solutions? How long do you leave in the steamer? How long do you cool down? Those are a few of the questions that I have so far......If there is any other helpful advice using this method it would be greatly appreciated. Thanks, Brandon Stokes, HT(ASCP) Lab Manager Cutting Edge Histology Services, LLC Tigard, OR 503-443-2157 --------------------------------- 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 From hhawkins <@t> UTMB.EDU Mon Nov 1 09:07:22 2004 From: hhawkins <@t> UTMB.EDU (Hawkins, Hal K.) Date: Fri Sep 16 15:24:14 2005 Subject: FW: [Histonet] Offtopic - Microscope for kids Message-ID: <8D6F233E2A5D574B929F3944F3316FD0F79EED@EXCH2K3.utmb.edu> Yes, maybe 12 is a better age to consider a professional-grade student scope. For an 8-yead-old, I would strongly recommend the digital microscopes from Intel, Mattel and Digital Blue, the QX3 and QX5. I have a QX3, and it is easy, lots of fun, and really an excellent way to view things in the macro range -- insects, plankton, money, color slides, nose hair -- whatever you like! Hal Hawkins -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of McCormick, James Sent: Saturday, October 30, 2004 8:34 AM To: John Kiernan; Kim Merriam; Histonet Subject: RE: [Histonet] Offtopic - Microscope for kids Kim, You have received some good advice from John Kiernan. Learning to use a hand lens to bring "little things" closer to view is the best building block for "experimenting" with the magnification of interesting objects. Someone has said "we need a low power lens and a high power mind!" There are some interesting books and web based assets to build with. From my experience it is important that the beginner will benefit greatly from "examining" little things with a hand lens and making drawings of what they see. If you look up "bugs" on Google it will lead you to www.BillyBear4Kids.com I think you will find some things of great interest to an 8 year old. Wait until he is 12 to spend $100.00 for a used student microscope. Good Luck, I am a pathologist and have used a microscope since age 12. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Sent: Saturday, October 30, 2004 12:48 AM To: Kim Merriam; Histonet Subject: Re: [Histonet] Offtopic - Microscope for kids WHATEVER YOU DO: Don't buy one without first having a look at something with it yourself. Even in the highest of the price ranges you mention the optics cannot be of high quality, so there's no point getting anything with an objective that's more than X10. The mechanical parts are also likely to be stiff, loose, or otherwise not easy to control. The best value for money is a 2nd-hand student microscope. University departments often sell these off for $100 or less, and you'd be lucky to get an equivalent new instrument for $1000. An old student microscope would not impress your 8 year-old son, because it would need too many adjustments (condenser, brightness, centering, positioning the slide etc). My experience with children and microscopes is that those younger than about 13 find it difficult to keep an eye steadily in place over an eyepiece - not too far away (nothing visible), not too close (disturbing images of reflected bent eyelashes), and not wobbling either. Children need help finding something to look at because moving the slide is another difficulty (especially if you don't have a mechanical stage, which is 2 more knobs). Focusing is a must, of course, and using coarse and fine knobs can easily be taught. >From my experiences with children (including me, my friends long ago, my 5 grown-up kids, and some of our grandchildren), you need to be 16 to handle a real microscope. Not everyone would agree. The Royal Microscopical Society (in Britain) has been campaigning for "a microscope in every school" for several years. "Every school" means elementary schools, which are for under-11s in the UK. The RMS has identified various kinds of microscope appropriate for children. They have a major web site, http://www.rms.org.uk Click on the Education tab, then on AMFES to get their advice on paedomicrophilia. Consider buying your son a X10 lens and showing him how to see a spider's fangs or the stinging hairs of a nettle. Hope this helps! John Kiernan London, Canada. -- ------------------------------- 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 _______________________________ Kim Merriam wrote: > > Hello all, > > My 8 year old son wants a microscope for Christmas. I have been searching on the internet for a decent one that does not cost an arm and a leg. It seems like there are lots of cheap ones in the $12-30 range and then some nice ones that are in the $100-200 range. Does anyone know where I could get one that costs around $50-70 that is not a piece of junk? I would like 2 or 3 objectives. Of course, the slides need not be included! > > Thanks in advance, > Kim Merriam > Novartis > Cambridge, MA > > Kim Merriam > Novartis > Cambridge, MA > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Mon Nov 1 09:23:41 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] p21 mAb Message-ID: Is anyone doing IHC for p21 (clone EA10) on paraffin sections? If so, where do you get it? Thank you! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From MAUGER <@t> email.chop.edu Mon Nov 1 10:03:04 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] p21 mAb Message-ID: Richard, Several years ago (5) I was using p21 (EA10) fromNovocastra. We were using microwave retrieval at the time, in citra buffer pH6. Incubated overnight at 1:100. Jo >>> "Richard Cartun" 11/01/04 10:23AM >>> Is anyone doing IHC for p21 (clone EA10) on paraffin sections? If so, where do you get it? 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 From Jackie.O'Connor <@t> abbott.com Mon Nov 1 10:33:37 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Bcl2 Message-ID: I've had some replies to this quest, but I would like some more opinions please - I'm using a mouse monoclonal Bcl2 antibody and a DAKO Envision polymer kit. My control is beautiful, however, my test tissue has a present but weaker signal. I'd appreciate some input as to how to amplify this weak signal without amplifying the background. So, everyone put down the Halloween candy you scammed from your kids Trick or Treat bags - and help me out! My kids are too old to go Trick or Treating - I don't have any candy! However, I did costume my 110 pound White German Shepherd as a cow. . . . Jackie O' From Rcartun <@t> harthosp.org Mon Nov 1 10:37:09 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] H. pylori control tissue Message-ID: Dear Colleagues: Back in August I sent an e-mail indicating that I had extra control tissue for Helicobacter pylori. I received an overwhelming number of requests for tissue blocks. I am now in the process of sending out blocks to those of you who e-mailed me a complete mailing address, telephone number, and a Federal Express account number for billing. I apologize for the delay, but its been difficult finding the time to do this. In addition, it took an "Act from Congress" to get Federal Express to send me shipping supplies since our hospital no longer uses FedEx for overnight deliveries. In any event, if you sent me the above information, your block should arrive in a day or two. If not, send me another e-mail with all the information listed above and I will try to send you a block since I still have some left. First come, first served. Thank you for your patience. 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 ploykasek <@t> phenopath.com Mon Nov 1 11:09:03 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] p21 mAb In-Reply-To: Message-ID: We have used p21, clone EA10 from Calbiochem, but this was several years ago. We have found that clone DCS-60.2 from Lab Vision. We have better, more reproducible staining with clone DCS-60.2. Let me know if you'd like titer, pretreatment info. Good luck with your staining. Patti Loykasek PhenoPath laboratories Seattle, WA> Is anyone doing IHC for p21 (clone EA10) on paraffin sections? If so, > where do you get it? 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 From jcline <@t> wchsys.org Mon Nov 1 12:33:53 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Brandon Stokes Message-ID: <001701c4c041$56bdc0e0$1d2a14ac@wchsys.org> I use a Black & Decker steamer. I use Trilogy by Cell Marque and I heat the steamer up for 15 minutes before placing the slides into the steamer. I steam for 20 minutes (I do not preheat the Trilogy) and cool the slides after removing them from the steamer for 10 minutes. Rinse 3X in distilled water. ***** 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 RITA.ANGEL <@t> UC.EDU Mon Nov 1 13:04:37 2004 From: RITA.ANGEL <@t> UC.EDU (Rita Angel) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] B & D steamer Message-ID: <5.1.0.14.2.20041101140154.00b2d490@ucmail3.uc.edu> Hi, Since we're on the topic of steamers, has anyone had problems with their glass coplin jars cracking, or do you only use plastic? I normally use the steamer for 20 minutes after it's been pre-heated and my glass jars cracked. Thanks, Rita Angel, HT (ASCP) From Rcartun <@t> harthosp.org Mon Nov 1 13:26:32 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Using expired reagents in IHC Message-ID: It has been brought to my attention that the CAP Laboratory checklist has been revised as of 9/30/04 and now includes a question, "Are all immunohistochemical reagents used within their indicated expiration dates?". I have always believed that it is acceptable to use expired regents (mainly primary antibodies) as long as their reactivity is acceptable and documented. Obviously, the expiration date that manufactures use is not an accurate reflection of the reagent's likelihood of poor performance. Does anyone have information on this change? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From ploykasek <@t> phenopath.com Mon Nov 1 13:27:12 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Antigen Retrieval w/ B&D Steamer..... In-Reply-To: <20041029233229.70857.qmail@web41407.mail.yahoo.com> Message-ID: Brandon ? I?ve tried to answer your questions. If you need more info, please contact me. Patti Loykasek PhenoPath Laboratories Seattle, WA > > How long do you pre-heat the solutions? We preheat the solutions for 15 sec- 2 min in the microwave, depending on the size of container (either plastic coplin jar or tissue tek container). Be careful not to superheat your solution & burn yourself! > How long do you leave in the steamer? We put the deparaffinized, hydrated sections into 95 degree solution & leave the slides in the steamer for either 20 or 30 minutes. Each antibody has a preferred pretreatment at either 20 or 30 minutes. We have several different solutions we use. Each antibody is titer checked with several different pretreatments & the best one becomes our SOP. We start the time when after adding the slides the pretreatment solution returns to 95 degrees (enlarge the holes in the steamer lid so that a thermometer will fit) - this brings some standardization to our pretreatments. > How long do you cool down? We cool down for 20 minutes, I have done 15 minutes at other places of employment. The most important thing is to allow for some slow cool down that is not immediate & thus not ?flash drying? to your slides. Hope this helps. > > Those are a few of the questions that I have so far......If there is any other > helpful advice using this method it would be greatly appreciated. > > Thanks, > > Brandon Stokes, HT(ASCP) > Lab Manager > Cutting Edge Histology Services, LLC > Tigard, OR > 503-443-2157 > > > --------------------------------- > 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 From michele.french <@t> bms.com Mon Nov 1 13:55:21 2004 From: michele.french <@t> bms.com (Michele French) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] IHC for Microglia Message-ID: <418694A9.6010508@bms.com> My colleague is trying to find an antibody that is specific for microglial cells that will work on formalin fixed, paraffin embedded mouse brain sections. Has any one had any luck with a particular antibody? From pzeitlow <@t> bbpllab.com Mon Nov 1 14:09:27 2004 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Using expired reagents in IHC Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A0BEAD9@bbplsrv1.bbpl> Not that many years ago, we went through the same thing with flow cytometry antibodies. I would assume that there will be no exceptions in a clinical setting, but would also welcome feedback. Pat -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Monday, November 01, 2004 1:27 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Using expired reagents in IHC It has been brought to my attention that the CAP Laboratory checklist has been revised as of 9/30/04 and now includes a question, "Are all immunohistochemical reagents used within their indicated expiration dates?". I have always believed that it is acceptable to use expired regents (mainly primary antibodies) as long as their reactivity is acceptable and documented. Obviously, the expiration date that manufactures use is not an accurate reflection of the reagent's likelihood of poor performance. Does anyone have information on this change? 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 From wkreider <@t> u.washington.edu Mon Nov 1 14:50:45 2004 From: wkreider <@t> u.washington.edu (Wayne Kreider) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] sample preparation/analysis to observe gas bubbles Message-ID: <4186A1A5.2050506@u.washington.edu> We're exploring a treatment in which small short-lived (perhaps on the order of seconds) gas bubbles may be generated in tissue. To this end, we're interested in learning the 'instantaneous' state of the tissue immediately after treatment with regard to the presence of any bubbles. Currently, our experiments utilize rabbit muscle. Are there any known histological sample preparation/analysis procedures that have been used to preserve/observe resident gas bubbles? We have considered a flash freezing followed by staining and/or electron microscopy. However, our expertise is not in histology, so we'd very much appreciate any suggestions and/or details regarding possible sample preparation. Thanks. Wayne -- /**********************/ /Wayne Kreider/ /University of Washington/ /Applied Physics Lab / CIMU/ /106 Old Fisheries/ /206-543-1324/ From SAllen <@t> exchange.hsc.mb.ca Mon Nov 1 15:01:08 2004 From: SAllen <@t> exchange.hsc.mb.ca (Sharon Allen) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Modified Gomori's Trichrome Message-ID: <304B5A264EC9974E8121B0EA14A9C3936619B9@hsc01mx1.hsc.mb.ca> Hi, I have been having some trouble getting the mitochondria to stain well with this stain. It stains only slightly when made up fresh. I have tried new reagents, low ph, No luck. Method I use: Mayer's Hematoxylin 5 min. wash Gomori's trichrome 10 min. wash 0.2% acetic acid 10 sec. Gomori's: 0.6 g chromotrope 2R 0.3 g fast green 0.6 g phosphotungstic acid 1.0 ml glacial acetic acid 100 ml H2O I just did the exact same method but used Celestine blue 2R for 5 min. (found an old method with this step in it) before the Mayer's Hematoxylin & the mitochondria stained up beautifully - bright red!! Can anyone explain why this would happen or have any trick to make this stain work without the celestine blue step. Thanks Sharon sallen@hsc.mb.ca This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. From browning <@t> HHSC.CA Mon Nov 1 15:16:43 2004 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] polyoma virus, SV40 Message-ID: <3AADFB88753AD31189C100902786B91C0E27851F@hch_nt_exchange.hhsc.ca> Is anyone out there using an antibody against polyoma virus, SV40; BK; or JC, against human tissue for demonstrating kidney rejection? If so, could you share the details, thanks. Debra Browning, ART Technical Specialist, Immunohistochemistry Anatomic Pathology Hamilton Health Sciences phone: (905) 527-4322 ext 46131 e-mail: browning@hhsc.ca fax: (905) 524-2681 This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From karrie.langdon <@t> sympatico.ca Mon Nov 1 15:31:23 2004 From: karrie.langdon <@t> sympatico.ca (Walker/Langdon) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] cracking jars Message-ID: <008f01c4c05a$24e58df0$8a19fea9@walkercvzdlx4z> Hi histonetters! In answer to Rita's question: I sure have had problems with cracking coplin jars. I'm trying to make a switch from microwave to steamer, but am being foiled by shattering jars. I've been using the same basic approach as Rita. Would love some imput, Carrie Langdon, Ph.D. Dept. Path. and Molecular Med. McMaster University Hamilton ON From LuckG <@t> empirehealth.org Mon Nov 1 15:32:58 2004 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] sample preparation/analysis to observe gas bubbles Message-ID: Wayne, You may want to explore the expertise of the dept of neuropathology at Harborview in your hometown. Two individuals in particular. One of the neuropathologists, Dr. Donald E. Born, M.D. and the lead neuropathology histotechnologist, Christiana, of who's specialties include muscle biopsies. One of their specimen prep techniques includes flash freezing muscle in 2-Methylbutane (cooled by liquid nitrogen) @ -160 degrees C. In theory this minimizes morphological distortion of the specimen by rapidly forcing the water component of the tissue to change from a liquid to a gas without passing through a solid state (my inorganic chemistry classes are way too far in the past to allow me to recall the name for this physical process), but thought this might have some applicability to or lead you down a new pathway in the work you're doing. Good luck, 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: Wayne Kreider [mailto:wkreider@u.washington.edu] Sent: Monday, November 01, 2004 12:51 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] sample preparation/analysis to observe gas bubbles We're exploring a treatment in which small short-lived (perhaps on the order of seconds) gas bubbles may be generated in tissue. To this end, we're interested in learning the 'instantaneous' state of the tissue immediately after treatment with regard to the presence of any bubbles. Currently, our experiments utilize rabbit muscle. Are there any known histological sample preparation/analysis procedures that have been used to preserve/observe resident gas bubbles? We have considered a flash freezing followed by staining and/or electron microscopy. However, our expertise is not in histology, so we'd very much appreciate any suggestions and/or details regarding possible sample preparation. Thanks. Wayne -- /**********************/ /Wayne Kreider/ /University of Washington/ /Applied Physics Lab / CIMU/ /106 Old Fisheries/ /206-543-1324/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Mon Nov 1 15:36:51 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] cracking jars Message-ID: I have found that if you use borosilicate glass jars, you get better mileage out of it. They are the ones with the round seal at the bottom of the jar, and are also green-yellowish in color. 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: Walker/Langdon [mailto:karrie.langdon@sympatico.ca] Sent: Monday, November 01, 2004 3:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cracking jars Hi histonetters! In answer to Rita's question: I sure have had problems with cracking coplin jars. I'm trying to make a switch from microwave to steamer, but am being foiled by shattering jars. I've been using the same basic approach as Rita. Would love some imput, Carrie Langdon, Ph.D. Dept. Path. and Molecular Med. McMaster University Hamilton ON _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Mon Nov 1 15:47:56 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Using expired reagents in IHC In-Reply-To: Message-ID: Dr. Cartun, even though I had a procedure stating that all expired antibodies will be tested, that the slide with the date and reactivity will be kept on file and the annotation will be placed on the specificity sheet, it was unacceptable when CLIA came through. We where using reagents beyond the expiration date that the manufacturer suggested and we were in violation. Even though I told this yahoo that my other procedures state that a known positive control will be used for each run and if the positive control did not work, a repeat would be performed. If the negative result was contributed by an expired reagent, then a new reagent would be used. I think this is bull because I'm throwing away thousands of $$$$$ in reagents. 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 Richard Cartun Sent: Monday, November 01, 2004 1:27 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Using expired reagents in IHC It has been brought to my attention that the CAP Laboratory checklist has been revised as of 9/30/04 and now includes a question, "Are all immunohistochemical reagents used within their indicated expiration dates?". I have always believed that it is acceptable to use expired regents (mainly primary antibodies) as long as their reactivity is acceptable and documented. Obviously, the expiration date that manufactures use is not an accurate reflection of the reagent's likelihood of poor performance. Does anyone have information on this change? 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 From jhabecke <@t> seattlecca.org Mon Nov 1 16:02:10 2004 From: jhabecke <@t> seattlecca.org (Randolph-Habecker, Julie) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] RE:IHC in mouse lung Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAD2B@wala01.seattlecca.org> When working with mouse lung and any other mouse tissue it is important to first block with a Avidin block and a Biotin block. This will bind the natural avidin/ biotin that can cause background and sticking on the tissue. In addition to this block you will want to do a serum protein block. Using a 15/5% dilution of normal serums. The 15%, should be from the same species your secondary was made in (i.e.. if your using Goat anti Mouse as your secondary-You would block with Goat serum). And the 5% dilution will be mouse serum. These two serums should be mixed together using antibody diluent. Block with the serum at least 10 minutes, rinse, then proceed to apply the antibody that you are using. I use this technique in my lab on all of our mouse tissue and it works great! So by taking these steps, it should clear up any background problems you may be having doing IHC in your mouse lung. Lisa Lisa McLemore Lead Histologist Experimental Histopathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave. N, G1-300 PO Box 19023 Seattle, WA 98109-1024 Tel: (206) 288-6898 FAX: (206) 288-1345 lmclemore@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 peoshel <@t> wisc.edu Mon Nov 1 16:31:31 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] sample preparation/analysis to observe gas bubbles In-Reply-To: <4186A1A5.2050506@u.washington.edu> References: <4186A1A5.2050506@u.washington.edu> Message-ID: Cryofixation is the only way I know to preserve this kind of structure. There have been discussions about this in the past, so you might want to poke around in the Histonet archives. What kind of treatment? Is it something that you stimulate with say an electrode? If so, there are plunge freezers designed to allow you to stimulate the sample while it is falling into the liquid nitrogen. Rapidity of freezing is essential to insure vitrification of the water, instead of crystal formation. Although some folks maintain that the water doesn't vitrify, but instead forms micro-crystallites small enough to have no effect on structure. Others state that evanescent microspherules are formed, sort of neither fish nor fowl -- not crystalline and not glass. Either way, the frozen samples have to be maintained below the recrystallization temperature (around -80deg C or so) until after dehydraton or embedding to prevent crystal growth and negating all the rapid freezing goodness. But, the 4 basic methods are high-pressure freezing, propane-jet freezing, slam-freezing, and plunging into cryogen. The first three will likely distort the air bubbles and tissue around them, however. Plunging works well, as long as the plunge is made very rapidly, and no time is spent hanging about in the cold -- well below freezing -- nitrogen atmosphere above the cryogen. Many people plunge into some organic fluid like ethane which is held in a container in liquid nitrogen. This works, but it has a few issues: the cryogen is usually warmer than the LN2, so the freezing is not as rapid as it needs to be, and such organic cryogens are flammable or explosive when they warm up. Especially since, being near LN2 temperatures, they're enriching themselves in oxygen from the air (liquid air is warmer than liquid nitrogen). They can be handled safely, but this does require some thought. A better way is to plunge into slush nitrogen -- LN2 near the freezing point, instead of near the boiling point. This is about 14 deg C colder than LN2, so freezes samples faster, and so gives a greater depth of good freezing. It's also nitrogen, so there's no flammable gas to deal with (nor bureaucrats upset about flammable gases in your lab). It's easy to produce: just put a beaker full in a small to medium size vacuum desiccator, and pull a vacuum with a high capacity pump, like a dual-stage rotary pump. One used to rough out a large EM column usually works. Phil >We're exploring a treatment in which small short-lived (perhaps on >the order of seconds) gas bubbles may be generated in tissue. To >this end, we're interested in learning the 'instantaneous' state of >the tissue immediately after treatment with regard to the presence >of any bubbles. Currently, our experiments utilize rabbit muscle. >Are there any known histological sample preparation/analysis >procedures that have been used to preserve/observe resident gas >bubbles? We have considered a flash freezing followed by staining >and/or electron microscopy. However, our expertise is not in >histology, so we'd very much appreciate any suggestions and/or >details regarding possible sample preparation. Thanks. >Wayne >-- > >/**********************/ > >/Wayne Kreider/ > >/University of Washington/ > >/Applied Physics Lab / CIMU/ > >/106 Old Fisheries/ > >/206-543-1324/ > >_______________________________________________ >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 scoop <@t> mail.nih.gov Mon Nov 1 16:39:41 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] IHC for Microglia In-Reply-To: <418694A9.6010508@bms.com> References: <418694A9.6010508@bms.com> Message-ID: There are two antibodies I know of that work on ffpe macrophages including microglia. They are ED-1 (Serotec) (requires pH 6.0 citrate antigen retrieval, I use a microwave pressure cooker) and Mac-3 (Pharmingen) (supposedly doesn't require antigen retrieval, hasn't been as strong staining as ED-1 but I haven't really tried that hard). They both stain golgi, so theoretically stain other cell types as well, but in reality only stain macrophages so you can see it and it seems to be well accepted in the literature that they are macrophage-specific markers. Sharon >My colleague is trying to find an antibody that is specific for >microglial cells that will work on formalin fixed, paraffin embedded >mouse brain sections. Has any one had any luck with a particular >antibody? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From portera203 <@t> yahoo.com Mon Nov 1 17:50:38 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] cracking jars In-Reply-To: <008f01c4c05a$24e58df0$8a19fea9@walkercvzdlx4z> Message-ID: <20041101235038.29444.qmail@web40913.mail.yahoo.com> Depending on the size of the steamer you are using, we have purchased Plastic (not sure exactly what their make up is) jars that hold 10 slides and we use tissue tek staining racks and clear dishes for large volume of slides. Supplier is Evergreen Scientific. Walker/Langdon wrote:Hi histonetters! In answer to Rita's question: I sure have had problems with cracking coplin jars. I'm trying to make a switch from microwave to steamer, but am being foiled by shattering jars. I've been using the same basic approach as Rita. Would love some imput, Carrie Langdon, Ph.D. Dept. Path. and Molecular Med. McMaster University Hamilton ON _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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!? Check out the new Yahoo! Front Page. www.yahoo.com/a From portera203 <@t> yahoo.com Mon Nov 1 17:56:44 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Using expired reagents in IHC In-Reply-To: Message-ID: <20041101235644.13521.qmail@web40909.mail.yahoo.com> I am curious if the negative responses from the inspection teams include primary antibodies that have been aliquoted and frozen. Do they consider those expired if they are past the expiration date suggested by the manufacturer? We are undergoing CAP inspection on Friday (5th) feed back would be appreciated. I think that it is crazy with all of the hoops the have us jumping through with documentation for testing and validating, that they would make us discard the $$$$$$$$ in primaries. Makes no sense to me either. I will be interested to see what happens with our inspection. Richard Cartun wrote:It has been brought to my attention that the CAP Laboratory checklist has been revised as of 9/30/04 and now includes a question, "Are all immunohistochemical reagents used within their indicated expiration dates?". I have always believed that it is acceptable to use expired regents (mainly primary antibodies) as long as their reactivity is acceptable and documented. Obviously, the expiration date that manufactures use is not an accurate reflection of the reagent's likelihood of poor performance. Does anyone have information on this change? 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 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!? Check out the new Yahoo! Front Page. www.yahoo.com/a From jnocito <@t> satx.rr.com Mon Nov 1 20:15:34 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Using expired reagents in IHC References: <20041101235644.13521.qmail@web40909.mail.yahoo.com> Message-ID: <008a01c4c081$d62ffd80$286ece44@yourxhtr8hvc4p> Amy, when I was at the Armed Forces Institute of Pathology (AFIP) in Washington D.C., we froze some antibodies in 1982 at -80 and were still using them when I left there in 1991. Sometimes we had to retiter the antibody because we think that keeping antibodies at the temp for a long time had a freeze-dried effect, which made the antibody more concentrated. That was before all these new regulations came into play. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Amy Porter" To: "Richard Cartun" ; Sent: Monday, November 01, 2004 5:56 PM Subject: Re: [Histonet] Using expired reagents in IHC > I am curious if the negative responses from the inspection teams include primary antibodies that have been aliquoted and frozen. Do they consider those expired if they are past the expiration date suggested by the manufacturer? We are undergoing CAP inspection on Friday (5th) feed back would be appreciated. I think that it is crazy with all of the hoops the have us jumping through with documentation for testing and validating, that they would make us discard the $$$$$$$$ in primaries. Makes no sense to me either. I will be interested to see what happens with our inspection. > > Richard Cartun wrote:It has been brought to my attention that the CAP Laboratory checklist > has been revised as of 9/30/04 and now includes a question, "Are all > immunohistochemical reagents used within their indicated expiration > dates?". I have always believed that it is acceptable to use expired > regents (mainly primary antibodies) as long as their reactivity is > acceptable and documented. Obviously, the expiration date that > manufactures use is not an accurate reflection of the reagent's > likelihood of poor performance. Does anyone have information on this > change? 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 > > > > 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!? > 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 wkreider <@t> u.washington.edu Tue Nov 2 02:16:41 2004 From: wkreider <@t> u.washington.edu (Wayne Kreider) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] sample preparation/analysis to observe gas bubbles In-Reply-To: References: <4186A1A5.2050506@u.washington.edu> Message-ID: <41874269.5060503@u.washington.edu> Phil, Thanks for the input. Unfortunately, I haven't had much luck looking in the archive. The treatment is actually acoustic (high intensity ultrasound), so no electrodes are involved. Our treatment volume that we'd ideally like to freeze is about 2mm x 2mm x 5mm. However, from the little I've read about plunging, a guideline for the maximum sample thickness is 0.2mm. Using a slush as you describe would presumably help some, but I suspect 2mm is still too thick. If we are able to obtain a suitably thin sample, is plunging literally just dropping the raw tissue into the cryogen? Or are there some additional steps that should be taken in handling the sample? Moreover, if we do have a larger sample, reference books suggest that it's necessary to use a cryo-protectant. Is a cryo-protectant likely to distort the tissue, in particular any bubble-type structures? Also, what time delays are typically associated with using a protectant? Any suggestions about which cryo-protectant might be best suited for this application? Thanks again... I'd appreciate any thoughts you might have. Wayne Philip Oshel wrote: > Cryofixation is the only way I know to preserve this kind of > structure. There have been discussions about this in the past, so you > might want to poke around in the Histonet archives. > What kind of treatment? Is it something that you stimulate with say an > electrode? > If so, there are plunge freezers designed to allow you to stimulate > the sample while it is falling into the liquid nitrogen. > Rapidity of freezing is essential to insure vitrification of the > water, instead of crystal formation. Although some folks maintain that > the water doesn't vitrify, but instead forms micro-crystallites small > enough to have no effect on structure. Others state that evanescent > microspherules are formed, sort of neither fish nor fowl -- not > crystalline and not glass. Either way, the frozen samples have to be > maintained below the recrystallization temperature (around -80deg C or > so) until after dehydraton or embedding to prevent crystal growth and > negating all the rapid freezing goodness. > But, the 4 basic methods are high-pressure freezing, propane-jet > freezing, slam-freezing, and plunging into cryogen. The first three > will likely distort the air bubbles and tissue around them, however. > Plunging works well, as long as the plunge is made very rapidly, and > no time is spent hanging about in the cold -- well below freezing -- > nitrogen atmosphere above the cryogen. > Many people plunge into some organic fluid like ethane which is held > in a container in liquid nitrogen. This works, but it has a few > issues: the cryogen is usually warmer than the LN2, so the freezing is > not as rapid as it needs to be, and such organic cryogens are > flammable or explosive when they warm up. Especially since, being near > LN2 temperatures, they're enriching themselves in oxygen from the air > (liquid air is warmer than liquid nitrogen). They can be handled > safely, but this does require some thought. > A better way is to plunge into slush nitrogen -- LN2 near the freezing > point, instead of near the boiling point. This is about 14 deg C > colder than LN2, so freezes samples faster, and so gives a greater > depth of good freezing. It's also nitrogen, so there's no flammable > gas to deal with (nor bureaucrats upset about flammable gases in your > lab). It's easy to produce: just put a beaker full in a small to > medium size vacuum desiccator, and pull a vacuum with a high capacity > pump, like a dual-stage rotary pump. One used to rough out a large EM > column usually works. > > Phil > >> We're exploring a treatment in which small short-lived (perhaps on >> the order of seconds) gas bubbles may be generated in tissue. To >> this end, we're interested in learning the 'instantaneous' state of >> the tissue immediately after treatment with regard to the presence of >> any bubbles. Currently, our experiments utilize rabbit muscle. Are >> there any known histological sample preparation/analysis procedures >> that have been used to preserve/observe resident gas bubbles? We >> have considered a flash freezing followed by staining and/or electron >> microscopy. However, our expertise is not in histology, so we'd very >> much appreciate any suggestions and/or details regarding possible >> sample preparation. Thanks. >> Wayne >> -- >> >> /**********************/ >> >> /Wayne Kreider/ >> >> /University of Washington/ >> >> /Applied Physics Lab / CIMU/ >> >> /106 Old Fisheries/ >> >> /206-543-1324/ >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- /**********************/ /Wayne Kreider/ /University of Washington/ /Applied Physics Lab / CIMU/ /106 Old Fisheries/ /206-543-1324/ From c.m.vanderloos <@t> amc.uva.nl Tue Nov 2 02:42:40 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] RE: p21 mAb Message-ID: <5881bc587ab0.587ab05881bc@amc.uva.nl> Dear Richard, For p21 WAF/CIP1 we used DakoCyto antibody M7202, clone SX118. After HIER (Tris-EDTA pH9.0, 15 min 100C) the antibody stained very well at 1:25 (60 min, RT) using an EnVision+ detection with DAB+ as chromogen. As "counterstain" we performed double staining with CD31 and alpha-actin to get an idea which nuclei actually stained. Results were great when using sequential double staining: p21 - EnV+ - DAB+ - CD31 or SMA - PowerVision-GAM/AP - Liquid Permanent Red (not too dark!!) - haematoxilin (weak!!). Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "Richard Cartun" Date Mon, 01 Nov 2004 10:23:41 -0500 To Subject [Histonet] p21 mAb Is anyone doing IHC for p21 (clone EA10) on paraffin sections? If so, where do you get it? Thank you! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From Andrew.Prior <@t> Smith-Nephew.com Tue Nov 2 03:38:23 2004 From: Andrew.Prior <@t> Smith-Nephew.com (Prior, Andrew) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Alcian Blue 8GX Message-ID: Does anyone know of a reliable and cheap supplier of Alcian Blue 8GX dry dye (50% dye content)? I was looking to re-order from Sigma but the price is exorbitant - over ?450 for 100g!!!!! That's over 5 times what we paid 3 years ago. Talk about inflation! (Unfortunately our budget isn't as big as say, the Red Sox) Hopefully there is another supplier out there who sells the dye at a reasonable price. Thanks in advance Andrew PS Hope all you Americans have voted - if not, why not?! Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. From Mandy <@t> serotec.co.uk Tue Nov 2 04:21:14 2004 From: Mandy <@t> serotec.co.uk (Mandy Townsend) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] IHC for Microglia Message-ID: Antibodies against the F4/80 antigen are well documented as recognising microglia in mouse. Serotec are able to supply an antibody to this antigen that may be of assistance as this is suitable for formalin fixed paraffin embedded tissue using Trypsin digestion pre-treatment. This antibody has been refered to many times on the histonet server, if you want to look back through archives for further recommendations. Do not hesitate to contact me or your local Serotec office/agent for further information. I hope this helps. Mandy Mandy Townsend MSc Technical Services Advisor Serotec Ltd 22 Bankside Station Approach Kidlington Oxfordshire OX5 1JE Tel: +44 1865 852733 Fax: +44 1865 852739 email: mandy@serotec.co.uk URL: http://www.serotec.com/ Serotec-Your first choice for antibodies! Join our free email update service 4 antibodies for the price of 3 during October & November. More details IMPORTANT NOTICE: This message and any attachments may be confidential. If this has been sent to you in error, please contact the sender as soon as possible. Serotec Ltd. Registered in England No.1604642 Registered Office: Boswell House, 1-5 Broad Street, Oxford, OX1 SAW. UK -----Original Message----- From: Sharon Cooperman [mailto:scoop@mail.nih.gov] Sent: Monday, November 01, 2004 10:40 PM To: Michele French Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IHC for Microglia There are two antibodies I know of that work on ffpe macrophages including microglia. They are ED-1 (Serotec) (requires pH 6.0 citrate antigen retrieval, I use a microwave pressure cooker) and Mac-3 (Pharmingen) (supposedly doesn't require antigen retrieval, hasn't been as strong staining as ED-1 but I haven't really tried that hard). They both stain golgi, so theoretically stain other cell types as well, but in reality only stain macrophages so you can see it and it seems to be well accepted in the literature that they are macrophage-specific markers. Sharon >My colleague is trying to find an antibody that is specific for >microglial cells that will work on formalin fixed, paraffin embedded >mouse brain sections. Has any one had any luck with a particular >antibody? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________ This e-mail has been scanned for all viruses by Star. The service is powered by MessageLabs. For more information on a proactive anti-virus service working around the clock, around the globe, visit: http://www.star.net.uk ________________________________________________________________________ From BMolinari <@t> heart.thi.tmc.edu Tue Nov 2 06:28:05 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Alcian Blue 8GX Message-ID: Hi Andrew, It looks like there may be a record turnout this year. Unfortunately it may be weeks before the official results are determined. There will probably be several lawsuits in key states and our overseas military will be given more time to send in their absentee voters. Just a note..the NY Yankees have the biggest budget. Betsy Molinari HT (ASCP) and a Red Sox fan Texas Heart Institute Houston, Texas 77090 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Prior, Andrew Sent: Tuesday, November 02, 2004 3:38 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Alcian Blue 8GX Does anyone know of a reliable and cheap supplier of Alcian Blue 8GX dry dye (50% dye content)? I was looking to re-order from Sigma but the price is exorbitant - over ?450 for 100g!!!!! That's over 5 times what we paid 3 years ago. Talk about inflation! (Unfortunately our budget isn't as big as say, the Red Sox) Hopefully there is another supplier out there who sells the dye at a reasonable price. Thanks in advance Andrew PS Hope all you Americans have voted - if not, why not?! Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Tue Nov 2 08:56:54 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] cracking jars In-Reply-To: <008f01c4c05a$24e58df0$8a19fea9@walkercvzdlx4z> Message-ID: <000001c4c0ec$31567120$4a737b81@Cygnus> Why does anyone use coplin jars to begin with??? Why not use a plastic reagent tub (I use Tissue-Tek green tubs, VWR 25608-904) and slide racks (VWR 25608-868)? The only down side I can see from this is the volume of reagent used (150 - 200 mL). But then again, what is wasted from cracked coplin jars or ruined slides due to boiled out buffers. I don't use microwave, but I do use pressure cookers and these products are excellent for HIER. Hope this helps. Connie McManus GET INFORMED -- GET OUT AND VOTE!! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Walker/Langdon Sent: Monday, November 01, 2004 2:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cracking jars Hi histonetters! In answer to Rita's question: I sure have had problems with cracking coplin jars. I'm trying to make a switch from microwave to steamer, but am being foiled by shattering jars. I've been using the same basic approach as Rita. Would love some imput, Carrie Langdon, Ph.D. Dept. Path. and Molecular Med. McMaster University Hamilton ON _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tthinnes <@t> scripps.edu Tue Nov 2 09:50:46 2004 From: tthinnes <@t> scripps.edu (Terri Thinnes) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] 1) steaming slides 2) expired reagents for IHC Message-ID: <000801c4c0f3$b8fc39a0$2ab88389@loskutoff4> 1) I use plastic slide mailers filled with the antigen retrieval solution (about 15 to 25 ml) and place it in a beaker. Neither one have cracked or melted in the steamer. 2) I have had a few experiences with reagents/kits failing before their expiration date so I regard the expiration date as a guide only. I sent the kit back to the supplier and they always "verified" the failure and sent me a new one but a lot of time and frustration was involved. From flemons <@t> bhset.org Tue Nov 2 10:01:18 2004 From: flemons <@t> bhset.org (Fran Lemons) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Sta-on Message-ID: Can someone out there give me info on where to purchase Sta-on water bath adhesive? I can't find it in any of my catalogues. Thanks in advance, Fran Walker Histology Technical Specialist Baptist Hospital of E. TN (Knoxville) From mbecker <@t> pathlabinc.com Tue Nov 2 10:11:26 2004 From: mbecker <@t> pathlabinc.com (Michele Becker) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Sta-on In-Reply-To: Message-ID: Surgipath Item# 03105 pint Item# 03107 gallon 800-225-3035 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Fran Lemons Sent: Tuesday, November 02, 2004 11:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sta-on Can someone out there give me info on where to purchase Sta-on water bath adhesive? I can't find it in any of my catalogues. Thanks in advance, Fran Walker Histology Technical Specialist Baptist Hospital of E. TN (Knoxville) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jtsonger <@t> vt.edu Tue Nov 2 10:51:57 2004 From: jtsonger <@t> vt.edu (Jill Songer) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Sta-on In-Reply-To: References: Message-ID: <6.0.0.22.0.20041102115109.01b93ab8@pop.vt.edu> Surgipath. 800-225-3035. Been using it for years. Love it. At 11:01 AM 11/2/2004, Fran Lemons wrote: >Can someone out there give me info on where to purchase Sta-on water bath >adhesive? I can't find it in any of my catalogues. >Thanks in advance, >Fran Walker >Histology Technical Specialist >Baptist Hospital of E. TN (Knoxville) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Jill Songer HT (ASCP) Histopathology Lab Virginia-Maryland Regional College of Veterinary Medicine Teaching Hospital Virginia Tech Blacksburg, VA 24061-0443 www.vetmed.vt.edu From David.Muskett <@t> RLC.NHS.UK Tue Nov 2 10:55:22 2004 From: David.Muskett <@t> RLC.NHS.UK (Muskett David) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Microtomy Tutorial[Scanned] Message-ID: <6AB79F0BA7C4914BA8DD8F5E0C21B9752E2602@AHEXMAIL01.xalderhey.com> Dear All I am currently preparing a tutorial about the history of microtomy for my trainee staff. Does anybody have any documents that they would be willing to share. Thanks in advance David David Muskett Chief Biomedical Scientist, Histology Royal Liverpool Children's NHS Trust Eaton Road Liverpool, UK L12 2AP Internal telephone x3615 Telephone (0151) 293 3656 Fax (0151) 293 3617 http://nhsald02/histopathology From pruegg <@t> ihctech.net Tue Nov 2 11:29:03 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] IHCRG list serve Message-ID: <001d01c4c101$72da8c70$83020a0a@IHCTech> We have been getting bouce's from the emails listed below for the NSH IHC Resource Group listserv . If you recognize the email and need to update your address with us please let me know so you can continue to receive the messages posted on our listserv for members of the IHCRG. Thank you, Patsy Ruegg lelaangel@aol.com ladylynnoct@aol.com Christine.Mech@pharma.novartis.com dgoodwin@chsnj.org Dburke@swmedctr.com DLONGWOODWARD@BICS.BWH.HARVARD.EDU virgilio.cruz@impath.com jwebbol@jpshealthnetwork.org DOROTHY.ANN.EDWARDS@PHARMACIA.COM From info <@t> instrumedics.com Tue Nov 2 11:46:42 2004 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] sample preparation/analysis to observe gas bubbles References: <4186A1A5.2050506@u.washington.edu> <41874269.5060503@u.washington.edu> Message-ID: <007d01c4c103$ee0a4690$6401a8c0@INSTRUMEDICS22> Wayne, You may interested in Gentle Jane (GJ) snap freezing. The details can be found on our web site www.instrumedics.com .Click on snap-freezing on the home page or the link on the bottom of each page. You will find the steps in the GJ process with photos. The method not only freezes the tissue at -196 Celsius with a heat extractor that has been stored in liquid nitrogen. The heat extractor is 3/4lb, highly polished chrome plated brass, a good conductor. Both low temperature and thermal exchange are important for small ice crystal growth. Bernice Instrumedics Bernice ----- Original Message ----- From: "Wayne Kreider" To: "Philip Oshel" Cc: Sent: Tuesday, November 02, 2004 3:16 AM Subject: Re: [Histonet] sample preparation/analysis to observe gas bubbles > Phil, > Thanks for the input. Unfortunately, I haven't had much luck looking in > the archive. > > The treatment is actually acoustic (high intensity ultrasound), so no > electrodes are involved. Our treatment volume that we'd ideally like to > freeze is about 2mm x 2mm x 5mm. However, from the little I've read > about plunging, a guideline for the maximum sample thickness is 0.2mm. > Using a slush as you describe would presumably help some, but I suspect > 2mm is still too thick. > > If we are able to obtain a suitably thin sample, is plunging literally > just dropping the raw tissue into the cryogen? Or are there some > additional steps that should be taken in handling the sample? Moreover, > if we do have a larger sample, reference books suggest that it's > necessary to use a cryo-protectant. Is a cryo-protectant likely to > distort the tissue, in particular any bubble-type structures? Also, > what time delays are typically associated with using a protectant? Any > suggestions about which cryo-protectant might be best suited for this > application? Thanks again... I'd appreciate any thoughts you might have. > Wayne > > Philip Oshel wrote: > > > Cryofixation is the only way I know to preserve this kind of > > structure. There have been discussions about this in the past, so you > > might want to poke around in the Histonet archives. > > What kind of treatment? Is it something that you stimulate with say an > > electrode? > > If so, there are plunge freezers designed to allow you to stimulate > > the sample while it is falling into the liquid nitrogen. > > Rapidity of freezing is essential to insure vitrification of the > > water, instead of crystal formation. Although some folks maintain that > > the water doesn't vitrify, but instead forms micro-crystallites small > > enough to have no effect on structure. Others state that evanescent > > microspherules are formed, sort of neither fish nor fowl -- not > > crystalline and not glass. Either way, the frozen samples have to be > > maintained below the recrystallization temperature (around -80deg C or > > so) until after dehydraton or embedding to prevent crystal growth and > > negating all the rapid freezing goodness. > > But, the 4 basic methods are high-pressure freezing, propane-jet > > freezing, slam-freezing, and plunging into cryogen. The first three > > will likely distort the air bubbles and tissue around them, however. > > Plunging works well, as long as the plunge is made very rapidly, and > > no time is spent hanging about in the cold -- well below freezing -- > > nitrogen atmosphere above the cryogen. > > Many people plunge into some organic fluid like ethane which is held > > in a container in liquid nitrogen. This works, but it has a few > > issues: the cryogen is usually warmer than the LN2, so the freezing is > > not as rapid as it needs to be, and such organic cryogens are > > flammable or explosive when they warm up. Especially since, being near > > LN2 temperatures, they're enriching themselves in oxygen from the air > > (liquid air is warmer than liquid nitrogen). They can be handled > > safely, but this does require some thought. > > A better way is to plunge into slush nitrogen -- LN2 near the freezing > > point, instead of near the boiling point. This is about 14 deg C > > colder than LN2, so freezes samples faster, and so gives a greater > > depth of good freezing. It's also nitrogen, so there's no flammable > > gas to deal with (nor bureaucrats upset about flammable gases in your > > lab). It's easy to produce: just put a beaker full in a small to > > medium size vacuum desiccator, and pull a vacuum with a high capacity > > pump, like a dual-stage rotary pump. One used to rough out a large EM > > column usually works. > > > > Phil > > > >> We're exploring a treatment in which small short-lived (perhaps on > >> the order of seconds) gas bubbles may be generated in tissue. To > >> this end, we're interested in learning the 'instantaneous' state of > >> the tissue immediately after treatment with regard to the presence of > >> any bubbles. Currently, our experiments utilize rabbit muscle. Are > >> there any known histological sample preparation/analysis procedures > >> that have been used to preserve/observe resident gas bubbles? We > >> have considered a flash freezing followed by staining and/or electron > >> microscopy. However, our expertise is not in histology, so we'd very > >> much appreciate any suggestions and/or details regarding possible > >> sample preparation. Thanks. > >> Wayne > >> -- > >> > >> /**********************/ > >> > >> /Wayne Kreider/ > >> > >> /University of Washington/ > >> > >> /Applied Physics Lab / CIMU/ > >> > >> /106 Old Fisheries/ > >> > >> /206-543-1324/ > >> > >> _______________________________________________ > >> Histonet mailing list > >> Histonet@lists.utsouthwestern.edu > >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > -- > > /**********************/ > > /Wayne Kreider/ > > /University of Washington/ > > /Applied Physics Lab / CIMU/ > > /106 Old Fisheries/ > > /206-543-1324/ > > > From MTitford <@t> aol.com Tue Nov 2 11:51:12 2004 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] History of microtechnique Message-ID: <25F82F56.4A42D588.00762DB1@aol.com> David Muskett asks about where to find information about the history of microtechnique. A good place to start is with "A history of microtechnique" by Brian Bracegirdle. The second edition is still available here in the States from Science Heritage Limited, Lincolnwood, IL a second good book is "History of staining" by George Clark and Frederick Kasten. 3ed edition, Williams & Wilkins, Baltimore & London. The second book is probably out of print but available on interlibrary loan. There are many other sources but you can get information overload in this area! Mike Titford USA Pathology Mobile AL From Sandra.Etheridge <@t> gems8.gov.bc.ca Tue Nov 2 11:57:12 2004 From: Sandra.Etheridge <@t> gems8.gov.bc.ca (Etheridge, Sandra AGF:EX) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] AFB Stain Message-ID: <2437188E40DE2947BD3AE43C87F3CB5F118E6E@stand.idir.bcgov> Hello, all, Does anyone have a current method for acid fast bacilli using auramine-rhodamine? Is this a fluorescent stain? Thanks in advance. Sandra Etheridge BC Ministry of Agriculture, Food & Fisheries Animal Health Center, Histology Abbotsford, BC Canada From contact <@t> excaliburpathology.com Tue Nov 2 12:06:53 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Alcian Blue and steamer containers Message-ID: <20041102180653.38389.qmail@web50308.mail.yahoo.com> Hello Andrew, yes I have already voted today :) You can buy a smaller amount of alcian blue at http://www.anatechltdusa.com/Catalog/catalogspecialstains.html I use plastic coplin jars in my B&D steamer for small quantities of slides and the larger tissue-tek when I have more. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From jengirl1014 <@t> yahoo.com Tue Nov 2 12:10:48 2004 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Re: Sta-On Message-ID: <20041102181048.98641.qmail@web60604.mail.yahoo.com> I purchased it through a company called Surgi-Path. They're the ones that make it. 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!? Check out the new Yahoo! Front Page. www.yahoo.com/a From RFORD <@t> HCMHCARES.ORG Tue Nov 2 12:11:38 2004 From: RFORD <@t> HCMHCARES.ORG (Ford, Rhonda) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Epidermal growth factor Message-ID: <684DF8CDE1FB6A428499DA65D753D12201F61789@JUPITER.HCMH.ORG> Does anyone know any institution(s) who perform this testing? From PatPatterson <@t> mhd.com Tue Nov 2 12:26:35 2004 From: PatPatterson <@t> mhd.com (Patterson, Pat) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Job posting - Dallas Message-ID: <293C7C19EFF7D611AE1A0002A53F81140CB43B0D@omega.mhd.com> Named one of the "Best Places to work 2004" by the Dallas Business Journal. Our dedicated team is very outspoken in their appreciation of the excellent clinical environment we foster and the ongoing commitment Methodist has made to patient satisfaction. Our dual focus on out patients and our employees is working - 2002 Methodist Health System earned the Gold Seal of Approval form the Joint Commission on Accreditation of Healthcare Organizations and more than 28 percent of our new hires come from employee referral! If excellent patient care is one of your priorities for career satisfaction, then you belong at Methodist. Join our Histology team at Methodist Dallas Medical Center. We're seeking a full-time Histology Technician on the day shift in our busy Anatomic Pathology Section. Job responsibilities include processing, embedding, sectioning, routine and special staining, assisting in gross room, frozen sectioning and computer entry into the MediTech lab information system. Experience with microwave technology would be an added plus. Requires HT/HTL ASCP registry or eligible. Minimum 2-year experience desired. Contact: Pat Patterson, Manager Phone: (214) 947-3538 Fax: (214) 947-3524 email: PatPatterson@mhd.com *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. From pruegg <@t> ihctech.net Tue Nov 2 12:33:43 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] thanks and aquamount In-Reply-To: Message-ID: <001501c4c10a$7b4e4fa0$83020a0a@IHCTech> Sharon, Aquamount will not harden, you should seal with clear nail polish to keep it permanently. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Cooperman Sent: Thursday, October 28, 2004 12:16 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] thanks and aquamount Dear Histonetters, Thanks to everyone for the great advice on bone decal and oil red o stain. I have one last question related to the oil red o stain - I just bought a bottle of aquamount to use when mounting the oil red o stained slides, but it didn't come with instructions (on the bottle it refers to instructions). Are there any special instructions for aquamount? Do I need to seal the slides with nail polish or does aquamount harden? Or, does anyone know how I can contact Lerner to get a copy of the instructions (I bought it from Fisher - Lerner doesn't answer the phone number I have for them). Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Tue Nov 2 12:58:15 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] AFB Stain In-Reply-To: <2437188E40DE2947BD3AE43C87F3CB5F118E6E@stand.idir.bcgov> Message-ID: Sandra, attached is our procedure for Truant's AFB. We purchase the staining kit from Infolab. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Etheridge, Sandra AGF:EX Sent: Tuesday, November 02, 2004 11:57 AM To: (histonet@lists.utsouthwestern.edu) Subject: [Histonet] AFB Stain Hello, all, Does anyone have a current method for acid fast bacilli using auramine-rhodamine? Is this a fluorescent stain? Thanks in advance. Sandra Etheridge BC Ministry of Agriculture, Food & Fisheries Animal Health Center, Histology Abbotsford, BC Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Tue Nov 2 13:19:49 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] History of microtechnique References: <25F82F56.4A42D588.00762DB1@aol.com> Message-ID: <4187DDD5.6F8C39E1@uwo.ca> There is also a chapter on the History of Staining by F. Kasten in the 10th edition of "Conn's Biological Stains." This book (2002) is still in print. ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 _______________________________ MTitford@aol.com wrote: > > David Muskett asks about where to find information about the history of microtechnique. > A good place to start is with "A history of microtechnique" by Brian Bracegirdle. The second edition is still available here in the States from Science Heritage Limited, Lincolnwood, IL > a second good book is "History of staining" by George Clark and Frederick Kasten. 3ed edition, Williams & Wilkins, Baltimore & London. The second book is probably out of print but available on interlibrary loan. > There are many other sources but you can get information overload in this area! > > Mike Titford > USA Pathology > Mobile AL From caroline.stott <@t> anatomy.otago.ac.nz Tue Nov 2 14:49:23 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Re: GMA sectioning Message-ID: <5.2.1.1.0.20041103094227.02076a10@anatomy.otago.ac.nz> Hi Ben, We have quite regularly cut GMA at 10, 20, 30 and 40 microns. We use a cotton bud to apply a little water to the face of the block, just gently rubbing it in. Also place some water on the glass knife as the section will come off easier. Then take a section and push it to the bottom of the water bowl/dish so it will flatten out. Then slide the sections onto the slide. Heres the tricky part.... We put a piece of filter paper on the bench then the slide on top, then another piece of wet filter paper on top of the slide (and sections) and use a roller (like a paint roller) applying a little pressure while rolling around 20-30 times over the slide and finally place on a hotplate (not flat) that is around 60 degrees C. Haven't had a problem. :) Hope that makes sense. Caroline Caroline Stott Histology Service Unit University of Otago PO Box 913 Dunedin, New Zealand Ph (03) 479 7152 Fax (03) 479 7136 From wkreider <@t> u.washington.edu Tue Nov 2 14:54:20 2004 From: wkreider <@t> u.washington.edu (Wayne Kreider) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] sample preparation/analysis to observe gas bubbles In-Reply-To: References: <4186A1A5.2050506@u.washington.edu> <41874269.5060503@u.washington.edu> Message-ID: <4187F3FC.3060400@u.washington.edu> Phil, Thanks again... all very helpful. I think we can now plan how we'll have the best chance at 'catching' transient bubbles. I'm not sure we'll have our techniques together for sampling and freezing the tissue in the very near term (this week), but I am hopeful we can do/try this. By the way, you mentioned that frozen samples are not useful for EM... how so? Perhaps due to subsequent fixing procedures? I'm relatively unfamiliar with EM although TEM had been suggested for this work. Wayne Philip Oshel wrote: > Wayne, > > You do have a problem ... > 2 X 2 X 2 mm is too big for any proper freezing method. Even > high-pressure freezing, which can in ideal conditions give the > greatest depth of good freezing, only does at best 500 microns. The > other methods do 200 microns or less. Plunging into LN2-cooled > cryogens will do a few 10s of microns, no more. They're fine for light > microscopy (assuming migration of molecules from holed membranes or > desiccation isn't an issue), but not EM. > The major source of freezing damage isn't really mechanical > ice-crystal growth and damage, but dehydration of cells from crystal > growth. > Plunging is literally dropping the sample into the cryogen, but not > really. By then I mean you *rapidly* plunge it in, not just let it > fall. By hand, as fast as possible. This is important in order to get > through the layer of cold nitrogen above the cryogen (whatever it is). > Some commercial plungers are designed to drop the sample into the > cryogen from enough height to pass through the cold gas layer quickly. > These are used for electrophysiology -- stimulate the sample then > drop, or stimulate during the drop, and therefore the sample is frozen > X msec after stimulation. > I can see doing this with ultrasonication, depending on the size, etc. > of the sonciation apparatus. > Cryoprotectants. Um. I don't think you can use these. The tissue has > to soak in the cryoprotectant -- often moving up a gradient -- in > order to get the cryoprotectant completely infiltrated into the > sample. This takes an hour or more -- maybe several hours? You'd > either have to assume nothing happens with the bubbles in the tissue > while sitting around infiltrating with cryoprotectant, or you'd have > to get the cryoprotectant into the tissue first, then sonicate and > pretend the cryoprotectant doesn't change the tissue properties and > therefore resulting bubbles. > I wouldn't believe either of those choices. > I think you'll need to do the experiment, then freeze, and use pieces > of tissue small enough to freeze properly. But! It's not entirely bad > -- only one dimension has to be =<200 microns -- 100 microns, really. > The other dimensions could be 2 mm. > So: 2 mm X 2 mm X 100 microns (even thinner would be better) should > work. Just hold the tissue by a corner, so that the cryogen has > maximal access to both faces of the thin side. And move the tissue > once in the slush LN2, don't hold it in one place -- keep fresh > cryogen contacting the sample. > Let's see -- oh. Insulate the container (and vacuum desiccator) as > best as possible, and work quickly, since you'll only have a few > minutes to freeze before the slush LN2 warms up to regular > almost-boiling LN2. > (I think Bal-Tec makes an apparatus for making slush LN2, and maybe > freezing with it, but you don't really need it, it just maybe makes > things easier.) > > Phil > > Phil, > Thanks for the input. Unfortunately, I haven't had much luck looking > in the archive. > The treatment is actually acoustic (high intensity ultrasound), so no > electrodes are involved. Our treatment volume that we'd ideally like > to freeze is about 2mm x 2mm x 5mm. However, from the little I've > read about plunging, a guideline for the maximum sample thickness is > 0.2mm. Using a slush as you describe would presumably help some, but > I suspect 2mm is still too thick. > If we are able to obtain a suitably thin sample, is plunging literally > just dropping the raw tissue into the cryogen? Or are there some > additional steps that should be taken in handling the sample? > Moreover, if we do have a larger sample, reference books suggest that > it's necessary to use a cryo-protectant. Is a cryo-protectant likely > to distort the tissue, in particular any bubble-type structures? > Also, what time delays are typically associated with using a > protectant? Any suggestions about which cryo-protectant might be best > suited for this application? Thanks again... I'd appreciate any > thoughts you might have. > Wayne > > Philip Oshel wrote: > > Cryofixation is the only way I know to preserve this kind of > structure. There have been discussions about this in the past, so you > might want to poke around in the Histonet archives. > What kind of treatment? Is it something that you stimulate with say an > electrode? > If so, there are plunge freezers designed to allow you to stimulate > the sample while it is falling into the liquid nitrogen. > Rapidity of freezing is essential to insure vitrification of the > water, instead of crystal formation. Although some folks maintain that > the water doesn't vitrify, but instead forms micro-crystallites small > enough to have no effect on structure. Others state that evanescent > microspherules are formed, sort of neither fish nor fowl -- not > crystalline and not glass. Either way, the frozen samples have to be > maintained below the recrystallization temperature (around -80deg C or > so) until after dehydraton or embedding to prevent crystal growth and > negating all the rapid freezing goodness. > But, the 4 basic methods are high-pressure freezing, propane-jet > freezing, slam-freezing, and plunging into cryogen. The first three > will likely distort the air bubbles and tissue around them, however. > Plunging works well, as long as the plunge is made very rapidly, and > no time is spent hanging about in the cold -- well below freezing -- > nitrogen atmosphere above the cryogen. > Many people plunge into some organic fluid like ethane which is held > in a container in liquid nitrogen. This works, but it has a few > issues: the cryogen is usually warmer than the LN2, so the freezing is > not as rapid as it needs to be, and such organic cryogens are > flammable or explosive when they warm up. Especially since, being near > LN2 temperatures, they're enriching themselves in oxygen from the air > (liquid air is warmer than liquid nitrogen). They can be handled > safely, but this does require some thought. > A better way is to plunge into slush nitrogen -- LN2 near the freezing > point, instead of near the boiling point. This is about 14 deg C > colder than LN2, so freezes samples faster, and so gives a greater > depth of good freezing. It's also nitrogen, so there's no flammable > gas to deal with (nor bureaucrats upset about flammable gases in your > lab). It's easy to produce: just put a beaker full in a small to > medium size vacuum desiccator, and pull a vacuum with a high capacity > pump, like a dual-stage rotary pump. One used to rough out a large EM > column usually works. > > Phil > > We're exploring a treatment in which small short-lived (perhaps on the > order of seconds) gas bubbles may be generated in tissue. To this > end, we're interested in learning the 'instantaneous' state of the > tissue immediately after treatment with regard to the presence of any > bubbles. Currently, our experiments utilize rabbit muscle. Are there > any known histological sample preparation/analysis procedures that > have been used to preserve/observe resident gas bubbles? We have > considered a flash freezing followed by staining and/or electron > microscopy. However, our expertise is not in histology, so we'd very > much appreciate any suggestions and/or details regarding possible > sample preparation. Thanks. > Wayne > -- /**********************/ /Wayne Kreider/ /University of Washington/ /Applied Physics Lab / CIMU/ /106 Old Fisheries/ /206-543-1324/ From tdobersztyn <@t> chmca.org Tue Nov 2 11:55:07 2004 From: tdobersztyn <@t> chmca.org (tdobersztyn@chmca.org) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Re: Polyoma Virus Message-ID: Debra- BK, Polyoma virus, can be demonstrated beautifully with Electron Microscopy. That is, if you have access to an EM lab. I am frequently processing renal core biopsies to r/o BK. If you would like assistance- let me know. Date: Mon, 1 Nov 2004 16:16:43 -0500 From: Browning Deb Subject: [Histonet] polyoma virus, SV40 To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <3AADFB88753AD31189C100902786B91C0E27851F@hch_nt_exchange.hhsc.ca> Content-Type: text/plain; charset="iso-8859-1" Is anyone out there using an antibody against polyoma virus, SV40; BK; or JC, against human tissue for demonstrating kidney rejection? If so, could you share the details, thanks. Debra Browning, ART Technical Specialist, Immunohistochemistry Anatomic Pathology Hamilton Health Sciences phone: (905) 527-4322 ext 46131 e-mail: browning@hhs Theresa R Dobersztyn HT ASCP Electron Microscopy Laboratory Department of Pathology Akron Children's Hospital 1 Perkins Square Akron, Ohio 44308 330-543-8279 From lpwenk <@t> sbcglobal.net Tue Nov 2 19:06:49 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] AFB Stain References: <2437188E40DE2947BD3AE43C87F3CB5F118E6E@stand.idir.bcgov> Message-ID: <022801c4c141$65f66ca0$6639d445@domainnotset.invalid> Check with your microbiology department. They are probably doing it already. One difference I've found between doing this procedure on smears vs. fixed/processed tissue - with smears, there is a 5 minute acid-alcohol decolorization. With fixed/processed/paraffin embedded tissue, reduce this to 2-3 seconds. The AFB cells walls have been compromised with the fixative and alcohols and xylene of the processing. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Etheridge, Sandra AGF:EX" To: Sent: Tuesday, November 02, 2004 12:57 PM Subject: [Histonet] AFB Stain > Hello, all, > > Does anyone have a current method for acid fast bacilli using > auramine-rhodamine? Is this a fluorescent stain? > > Thanks in advance. > > Sandra Etheridge > > BC Ministry of Agriculture, Food & Fisheries > Animal Health Center, Histology > Abbotsford, BC > Canada > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From marshall <@t> cormack.uct.ac.za Wed Nov 3 04:33:41 2004 From: marshall <@t> cormack.uct.ac.za (Marshall) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Merkel cell staining Message-ID: <4188B405.A7723577@cormack.uct.ac.za> Hi all I am looking to demonstrate Merkel cells in skin. I can not find any reference in my textbooks to their demonstration although I have not used the internet. Also I have tried silver stains for demoing free nerve endings in skin with not much success. The immunocytochemistry for neuro filament protein was a bit better but not great. Is there anyway I can improve on this? From prippstein <@t> ottawaheart.ca Wed Nov 3 08:41:53 2004 From: prippstein <@t> ottawaheart.ca (Peter Rippstein) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] LR White vs LR Gold Message-ID: Hello Colleagues Have any of you had the opportunity to compare EM immunogold labelling efficiency of LR White vs LR Gold acrylic resins? Protocols would be helpful. Many thanks. Peter Peter Rippstein ART, MLT University of Ottawa Heart Institute 40 Ruskin St., Room H453A Ottawa, Ontario Canada, K1Y 4W7 Tel: (613) 761-5282 Fax: (613) 761-5281 email: prippstein@ottawaheart.ca -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Peter Rippstein EMAIL;WORK;PREF;NGW:PRippstein@ottawaheart.ca N:Rippstein;Peter END:VCARD From SAllen <@t> exchange.hsc.mb.ca Wed Nov 3 08:54:40 2004 From: SAllen <@t> exchange.hsc.mb.ca (Sharon Allen) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] RE: Modified Gomori's Trichrome Message-ID: <304B5A264EC9974E8121B0EA14A9C3936619BA@hsc01mx1.hsc.mb.ca> Hi Bruce, I know the Gomori's Trichrome isn't specific for mitochondria but it should stain red if the test is working properly. I have been doing this stain routinely for 4 different Neuropathologists & am now doing the muscles for another Neuropathologist who is apparently an expert in the field & extremely fussy. They all are of the same opinion. Having the mitochondria stained seems to be imperative for an acceptable stain. Using yet another stain on the muscle bx's is not an option. This is why I am attempting to find the key to the "perfect Gomori's Trichrome stain". Using the celestine blue before the Mayer's has worked well for staining the mitochondria (I have done 40 of them), but I have no explanation why. My theory is very rusty & hoped someone with more knowledge than I could explain the reason to me. I think it may have something to do with Celestine Blue staining DNA & being a oxazine dye. Also chromoptrope 2R being a metachromatic stain. I would appreciate any help you can give me. Thanks for your reply, Sharon -----Original Message----- This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. From alkozer <@t> piadc.ars.usda.gov Wed Nov 3 09:32:03 2004 From: alkozer <@t> piadc.ars.usda.gov (Amy Kozer) Date: Fri Sep 16 15:24:14 2005 Subject: [Histonet] Envision System with Vector Red Background Message-ID: I've tried using Vector Red with Envision system for permanance and have encountered a lot of background and precipitate. I've been staining frozen sections of epithelium fixed with acetone. Has anyone had any experience with this? Remedies? Thank you Amy Kozer PIADC, ARS, USDA From Rcartun <@t> harthosp.org Wed Nov 3 09:52:46 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Merkel cell staining Message-ID: Try IHC for cytokeratin 20. 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 >>> Marshall 11/03/04 05:33AM >>> Hi all I am looking to demonstrate Merkel cells in skin. I can not find any reference in my textbooks to their demonstration although I have not used the internet. Also I have tried silver stains for demoing free nerve endings in skin with not much success. The immunocytochemistry for neuro filament protein was a bit better but not great. Is there anyway I can improve on this? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RITA.ANGEL <@t> UC.EDU Wed Nov 3 10:09:31 2004 From: RITA.ANGEL <@t> UC.EDU (Rita Angel) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] B & D steamer Message-ID: <5.1.0.14.2.20041103110800.00b24e08@ucmail3.uc.edu> Thanks to everyone for their input on the type of containers you use in the steamer. I'll try the plastic coplin jars!! Thanks again, Rita Angel HT (ASCP) From sluhisto <@t> yahoo.com Wed Nov 3 10:52:46 2004 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Polyoma BK virus controls Message-ID: <20041103165246.4952.qmail@web51001.mail.yahoo.com> Does anyone have BK virus controls that you would be willing to trade for something we may have that you would need? Any help in locating control blocks or slides would be greatly appreciated. Thanks. Susan --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com/a From DRG <@t> Stowers-Institute.org Wed Nov 3 11:46:59 2004 From: DRG <@t> Stowers-Institute.org (Grant, Debra) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] BMP7 Message-ID: <1112618780-192831059@pathology.swmed.edu> Has anyone used BMP7 antibody on mouse? If so, could you please send me all the information, company, catalog #, protocol etc. Thanks in advance! Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org From neuroant <@t> hotmail.com Wed Nov 3 11:52:01 2004 From: neuroant <@t> hotmail.com (Ant S.) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Thanks for the Teasing Message-ID: Thanks for the great tips on Nerve Teasing. I have quite a few good ideas to experiment with now. I appreciate the help. This is a very informative community (and mostly cheerful, too!) Thanks, Antoinette Swensson Univeristy of Washington/Harborview Medical Center Neuropathology _________________________________________________________________ [1]Find the music you love on MSN Music. Start downloading now! References 1. http://g.msn.com/8HMBENUS/2731??PS=47575 From Rachael_Emerson <@t> URMC.Rochester.edu Wed Nov 3 13:00:53 2004 From: Rachael_Emerson <@t> URMC.Rochester.edu (Emerson, Rachael) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] CD41, GPV, GP1bBeta Message-ID: Hi. Does anyone have any experience working with CD41, GPV, or GP1bBeta antibodies? I could really use some help. Thanks a lot!! Rachael Emerson 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: histonet-request@lists.utsouthwestern.edu > Reply To: histonet@lists.utsouthwestern.edu > Sent: Wednesday, November 3, 2004 1:03 PM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet Digest, Vol 12, Issue 4 > > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. Alcian Blue and steamer containers (Paula Pierce) > 2. Re: Sta-On (Jennifer Sipes) > 3. Epidermal growth factor (Ford, Rhonda) > 4. Job posting - Dallas (Patterson, Pat) > 5. RE: thanks and aquamount (Patsy Ruegg) > 6. RE: AFB Stain (Joe Nocito) > 7. Re: History of microtechnique (John Kiernan) > 8. Re: GMA sectioning (Caroline Stott) > 9. Re: sample preparation/analysis to observe gas bubbles > (Wayne Kreider) > 10. Re: Polyoma Virus (tdobersztyn@chmca.org) > 11. Re: AFB Stain (lpwenk@sbcglobal.net) > 12. Merkel cell staining (Marshall) > 13. LR White vs LR Gold (Peter Rippstein) > 14. RE: Modified Gomori's Trichrome (Sharon Allen) > 15. Envision System with Vector Red Background (Amy Kozer) > 16. Re: Merkel cell staining (Richard Cartun) > 17. B & D steamer (Rita Angel) > 18. Polyoma BK virus controls (Histology SLU) > 19. BMP7 (Grant, Debra) > 20. Thanks for the Teasing (Ant S.) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Tue, 2 Nov 2004 10:06:53 -0800 (PST) > From: Paula Pierce > Subject: [Histonet] Alcian Blue and steamer containers > To: histonet@lists.utsouthwestern.edu > Message-ID: <20041102180653.38389.qmail@web50308.mail.yahoo.com> > Content-Type: text/plain; charset=us-ascii > > Hello Andrew, > > yes I have already voted today :) > > You can buy a smaller amount of alcian blue at > http://www.anatechltdusa.com/Catalog/catalogspecialstains.html > > I use plastic coplin jars in my B&D steamer for small quantities of slides > and the larger tissue-tek when I have more. > > > 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, 2 Nov 2004 10:10:48 -0800 (PST) > From: Jennifer Sipes > Subject: [Histonet] Re: Sta-On > To: histonet@lists.utsouthwestern.edu > Message-ID: <20041102181048.98641.qmail@web60604.mail.yahoo.com> > Content-Type: text/plain; charset=us-ascii > > I purchased it through a company called Surgi-Path. They're the ones that > make it. > > > 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!? > Check out the new Yahoo! Front Page. www.yahoo.com/a > > ------------------------------ > > Message: 3 > Date: Tue, 2 Nov 2004 13:11:38 -0500 > From: "Ford, Rhonda" > Subject: [Histonet] Epidermal growth factor > To: > Message-ID: > <684DF8CDE1FB6A428499DA65D753D12201F61789@JUPITER.HCMH.ORG> > Content-Type: text/plain; charset="us-ascii" > > Does anyone know any institution(s) who perform this testing? > > > ------------------------------ > > Message: 4 > Date: Tue, 2 Nov 2004 12:26:35 -0600 > From: "Patterson, Pat" > Subject: [Histonet] Job posting - Dallas > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: <293C7C19EFF7D611AE1A0002A53F81140CB43B0D@omega.mhd.com> > Content-Type: text/plain; charset="iso-8859-1" > > > Named one of the "Best Places to work 2004" by the Dallas Business > Journal. > > Our dedicated team is very outspoken in their appreciation of the > excellent > clinical environment we foster and the ongoing commitment Methodist has > made > to patient satisfaction. Our dual focus on out patients and our employees > is working - 2002 Methodist Health System earned the Gold Seal of > Approval > form the Joint Commission on Accreditation of Healthcare Organizations and > more than 28 percent of our new hires come from employee referral! If > excellent patient care is one of your priorities for career satisfaction, > then you belong at Methodist. > > > > Join our Histology team at Methodist Dallas Medical Center. We're seeking > a > full-time Histology Technician on the day shift in our busy Anatomic > Pathology Section. Job responsibilities include processing, embedding, > sectioning, routine and special staining, assisting in gross room, frozen > sectioning and computer entry into the MediTech lab information system. > Experience with microwave technology would be an added plus. Requires > HT/HTL > ASCP registry or eligible. Minimum 2-year experience desired. > > Contact: Pat Patterson, Manager > Phone: (214) 947-3538 > Fax: (214) 947-3524 > email: PatPatterson@mhd.com > > > *********************************************************************** > > This electronic transmission contains information from Methodist Health > System and should be considered confidential and privileged. The > information contained in the above messages is intended only for the > use of the individual(s) and entity(ies) named above. If you are not > the intended recipient, be aware that any disclosure, copying, > distribution, or use of this information is prohibited. If you receive > this transmission in error, please notify the sender immediately by > return e-mail. Methodist Health System, its subsidiaries and > affiliates hereby claim all applicable privileges related to the > transmission of this communication. > > > > ------------------------------ > > Message: 5 > Date: Tue, 2 Nov 2004 11:33:43 -0700 > From: "Patsy Ruegg" > Subject: RE: [Histonet] thanks and aquamount > To: "'Sharon Cooperman'" , > > Message-ID: <001501c4c10a$7b4e4fa0$83020a0a@IHCTech> > Content-Type: text/plain; charset="us-ascii" > > Sharon, > Aquamount will not harden, you should seal with clear nail polish to > keep it permanently. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon > Cooperman > Sent: Thursday, October 28, 2004 12:16 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] thanks and aquamount > > > Dear Histonetters, > > Thanks to everyone for the great advice on bone decal and oil red o > stain. I have one last question related to the oil red o stain - I > just bought a bottle of aquamount to use when mounting the oil red o > stained slides, but it didn't come with instructions (on the bottle > it refers to instructions). Are there any special instructions for > aquamount? Do I need to seal the slides with nail polish or does > aquamount harden? Or, does anyone know how I can contact Lerner to > get a copy of the instructions (I bought it from Fisher - Lerner > doesn't answer the phone number I have for them). > > Thanks, > Sharon > -- > Sharon Cooperman > NIH, NICHD, CBMB 301.435-8417 > Building 18T, room 101 301.402-0078 fax > Bethesda, MD 20892 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 6 > Date: Tue, 2 Nov 2004 12:58:15 -0600 > From: "Joe Nocito" > Subject: RE: [Histonet] AFB Stain > To: "Etheridge, Sandra AGF:EX" , > > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > Sandra, > attached is our procedure for Truant's AFB. We purchase the staining kit > from Infolab. > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of > Etheridge, Sandra AGF:EX > Sent: Tuesday, November 02, 2004 11:57 AM > To: (histonet@lists.utsouthwestern.edu) > Subject: [Histonet] AFB Stain > > > Hello, all, > > Does anyone have a current method for acid fast bacilli using > auramine-rhodamine? Is this a fluorescent stain? > > Thanks in advance. > > Sandra Etheridge > > BC Ministry of Agriculture, Food & Fisheries > Animal Health Center, Histology > Abbotsford, BC > Canada > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ > > Message: 7 > Date: Tue, 02 Nov 2004 14:19:49 -0500 > From: John Kiernan > Subject: Re: [Histonet] History of microtechnique > To: MTitford@aol.com > Cc: Histonet@lists.utsouthwestern.edu > Message-ID: <4187DDD5.6F8C39E1@uwo.ca> > Content-Type: text/plain; charset=us-ascii > > There is also a chapter on the History of Staining > by F. Kasten in the 10th edition of "Conn's > Biological Stains." This book (2002) is > still in print. > ------------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > _______________________________ > MTitford@aol.com wrote: > > > > David Muskett asks about where to find information about the history of > microtechnique. > > A good place to start is with "A history of microtechnique" by Brian > Bracegirdle. The second edition is still available here in the States from > Science Heritage Limited, Lincolnwood, IL > > a second good book is "History of staining" by George Clark and > Frederick Kasten. 3ed edition, Williams & Wilkins, Baltimore & London. The > second book is probably out of print but available on interlibrary loan. > > There are many other sources but you can get information overload in > this area! > > > > Mike Titford > > USA Pathology > > Mobile AL > > > > ------------------------------ > > Message: 8 > Date: Wed, 03 Nov 2004 09:49:23 +1300 > From: Caroline Stott > Subject: [Histonet] Re: GMA sectioning > To: Histonet@lists.utsouthwestern.edu > Message-ID: <5.2.1.1.0.20041103094227.02076a10@anatomy.otago.ac.nz> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Hi Ben, > We have quite regularly cut GMA at 10, 20, 30 and 40 microns. We use a > cotton bud to apply a little water to the face of the block, just gently > rubbing it in. Also place some water on the glass knife as the section > will come off easier. Then take a section and push it to the bottom of > the > water bowl/dish so it will flatten out. Then slide the sections onto the > slide. Heres the tricky part.... We put a piece of filter paper on the > bench then the slide on top, then another piece of wet filter paper on top > > of the slide (and sections) and use a roller (like a paint roller) > applying a little pressure while rolling around 20-30 times over the slide > > and finally place on a hotplate (not flat) that is around 60 degrees > C. Haven't had a problem. :) > Hope that makes sense. > > Caroline > > Caroline Stott > > Histology Service Unit > University of Otago > PO Box 913 > Dunedin, New Zealand > Ph (03) 479 7152 > Fax (03) 479 7136 > > ------------------------------ > > Message: 9 > Date: Tue, 02 Nov 2004 12:54:20 -0800 > From: Wayne Kreider > Subject: Re: [Histonet] sample preparation/analysis to observe gas > bubbles > To: Philip Oshel > Cc: histonet@lists.utsouthwestern.edu > Message-ID: <4187F3FC.3060400@u.washington.edu> > Content-Type: text/plain; charset=ISO-8859-1; format=flowed > > Phil, > Thanks again... all very helpful. I think we can now plan how we'll > have the best chance at 'catching' transient bubbles. I'm not sure > we'll have our techniques together for sampling and freezing the tissue > in the very near term (this week), but I am hopeful we can do/try this. > By the way, you mentioned that frozen samples are not useful for EM... > how so? Perhaps due to subsequent fixing procedures? I'm relatively > unfamiliar with EM although TEM had been suggested for this work. > Wayne > > Philip Oshel wrote: > > > Wayne, > > > > You do have a problem ... > > 2 X 2 X 2 mm is too big for any proper freezing method. Even > > high-pressure freezing, which can in ideal conditions give the > > greatest depth of good freezing, only does at best 500 microns. The > > other methods do 200 microns or less. Plunging into LN2-cooled > > cryogens will do a few 10s of microns, no more. They're fine for light > > microscopy (assuming migration of molecules from holed membranes or > > desiccation isn't an issue), but not EM. > > The major source of freezing damage isn't really mechanical > > ice-crystal growth and damage, but dehydration of cells from crystal > > growth. > > Plunging is literally dropping the sample into the cryogen, but not > > really. By then I mean you *rapidly* plunge it in, not just let it > > fall. By hand, as fast as possible. This is important in order to get > > through the layer of cold nitrogen above the cryogen (whatever it is). > > Some commercial plungers are designed to drop the sample into the > > cryogen from enough height to pass through the cold gas layer quickly. > > These are used for electrophysiology -- stimulate the sample then > > drop, or stimulate during the drop, and therefore the sample is frozen > > X msec after stimulation. > > I can see doing this with ultrasonication, depending on the size, etc. > > of the sonciation apparatus. > > Cryoprotectants. Um. I don't think you can use these. The tissue has > > to soak in the cryoprotectant -- often moving up a gradient -- in > > order to get the cryoprotectant completely infiltrated into the > > sample. This takes an hour or more -- maybe several hours? You'd > > either have to assume nothing happens with the bubbles in the tissue > > while sitting around infiltrating with cryoprotectant, or you'd have > > to get the cryoprotectant into the tissue first, then sonicate and > > pretend the cryoprotectant doesn't change the tissue properties and > > therefore resulting bubbles. > > I wouldn't believe either of those choices. > > I think you'll need to do the experiment, then freeze, and use pieces > > of tissue small enough to freeze properly. But! It's not entirely bad > > -- only one dimension has to be =<200 microns -- 100 microns, really. > > The other dimensions could be 2 mm. > > So: 2 mm X 2 mm X 100 microns (even thinner would be better) should > > work. Just hold the tissue by a corner, so that the cryogen has > > maximal access to both faces of the thin side. And move the tissue > > once in the slush LN2, don't hold it in one place -- keep fresh > > cryogen contacting the sample. > > Let's see -- oh. Insulate the container (and vacuum desiccator) as > > best as possible, and work quickly, since you'll only have a few > > minutes to freeze before the slush LN2 warms up to regular > > almost-boiling LN2. > > (I think Bal-Tec makes an apparatus for making slush LN2, and maybe > > freezing with it, but you don't really need it, it just maybe makes > > things easier.) > > > > Phil > > > > Phil, > > Thanks for the input. Unfortunately, I haven't had much luck looking > > in the archive. > > The treatment is actually acoustic (high intensity ultrasound), so no > > electrodes are involved. Our treatment volume that we'd ideally like > > to freeze is about 2mm x 2mm x 5mm. However, from the little I've > > read about plunging, a guideline for the maximum sample thickness is > > 0.2mm. Using a slush as you describe would presumably help some, but > > I suspect 2mm is still too thick. > > If we are able to obtain a suitably thin sample, is plunging literally > > just dropping the raw tissue into the cryogen? Or are there some > > additional steps that should be taken in handling the sample? > > Moreover, if we do have a larger sample, reference books suggest that > > it's necessary to use a cryo-protectant. Is a cryo-protectant likely > > to distort the tissue, in particular any bubble-type structures? > > Also, what time delays are typically associated with using a > > protectant? Any suggestions about which cryo-protectant might be best > > suited for this application? Thanks again... I'd appreciate any > > thoughts you might have. > > Wayne > > > > Philip Oshel wrote: > > > > Cryofixation is the only way I know to preserve this kind of > > structure. There have been discussions about this in the past, so you > > might want to poke around in the Histonet archives. > > What kind of treatment? Is it something that you stimulate with say an > > electrode? > > If so, there are plunge freezers designed to allow you to stimulate > > the sample while it is falling into the liquid nitrogen. > > Rapidity of freezing is essential to insure vitrification of the > > water, instead of crystal formation. Although some folks maintain that > > the water doesn't vitrify, but instead forms micro-crystallites small > > enough to have no effect on structure. Others state that evanescent > > microspherules are formed, sort of neither fish nor fowl -- not > > crystalline and not glass. Either way, the frozen samples have to be > > maintained below the recrystallization temperature (around -80deg C or > > so) until after dehydraton or embedding to prevent crystal growth and > > negating all the rapid freezing goodness. > > But, the 4 basic methods are high-pressure freezing, propane-jet > > freezing, slam-freezing, and plunging into cryogen. The first three > > will likely distort the air bubbles and tissue around them, however. > > Plunging works well, as long as the plunge is made very rapidly, and > > no time is spent hanging about in the cold -- well below freezing -- > > nitrogen atmosphere above the cryogen. > > Many people plunge into some organic fluid like ethane which is held > > in a container in liquid nitrogen. This works, but it has a few > > issues: the cryogen is usually warmer than the LN2, so the freezing is > > not as rapid as it needs to be, and such organic cryogens are > > flammable or explosive when they warm up. Especially since, being near > > LN2 temperatures, they're enriching themselves in oxygen from the air > > (liquid air is warmer than liquid nitrogen). They can be handled > > safely, but this does require some thought. > > A better way is to plunge into slush nitrogen -- LN2 near the freezing > > point, instead of near the boiling point. This is about 14 deg C > > colder than LN2, so freezes samples faster, and so gives a greater > > depth of good freezing. It's also nitrogen, so there's no flammable > > gas to deal with (nor bureaucrats upset about flammable gases in your > > lab). It's easy to produce: just put a beaker full in a small to > > medium size vacuum desiccator, and pull a vacuum with a high capacity > > pump, like a dual-stage rotary pump. One used to rough out a large EM > > column usually works. > > > > Phil > > > > We're exploring a treatment in which small short-lived (perhaps on the > > order of seconds) gas bubbles may be generated in tissue. To this > > end, we're interested in learning the 'instantaneous' state of the > > tissue immediately after treatment with regard to the presence of any > > bubbles. Currently, our experiments utilize rabbit muscle. Are there > > any known histological sample preparation/analysis procedures that > > have been used to preserve/observe resident gas bubbles? We have > > considered a flash freezing followed by staining and/or electron > > microscopy. However, our expertise is not in histology, so we'd very > > much appreciate any suggestions and/or details regarding possible > > sample preparation. Thanks. > > Wayne > > -- > > > /**********************/ > > /Wayne Kreider/ > > /University of Washington/ > > /Applied Physics Lab / CIMU/ > > /106 Old Fisheries/ > > /206-543-1324/ > > > > ------------------------------ > > Message: 10 > Date: Tue, 2 Nov 2004 12:55:07 -0500 > From: tdobersztyn@chmca.org > Subject: [Histonet] Re: Polyoma Virus > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset=US-ASCII > > > > > > > > > Debra- > BK, Polyoma virus, can be demonstrated beautifully with Electron > Microscopy. > That is, if you have access to an EM lab. > I am frequently processing renal core biopsies to r/o BK. > If you would like assistance- > let me know. > > > > Date: Mon, 1 Nov 2004 16:16:43 -0500 > From: Browning Deb > Subject: [Histonet] polyoma virus, SV40 > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > > <3AADFB88753AD31189C100902786B91C0E27851F@hch_nt_exchange.hhsc.ca> > Content-Type: text/plain; charset="iso-8859-1" > > Is anyone out there using an antibody against polyoma virus, SV40; BK; or > JC, against human tissue for demonstrating kidney rejection? If so, could > you share the details, thanks. > > Debra Browning, ART > Technical Specialist, Immunohistochemistry > Anatomic Pathology > Hamilton Health Sciences > phone: (905) 527-4322 ext 46131 > e-mail: browning@hhs > > > > > > Theresa R Dobersztyn HT ASCP > Electron Microscopy Laboratory > Department of Pathology > > Akron Children's Hospital > 1 Perkins Square > Akron, Ohio 44308 > > 330-543-8279 > > > > > ------------------------------ > > Message: 11 > Date: Tue, 2 Nov 2004 20:06:49 -0500 > From: > Subject: Re: [Histonet] AFB Stain > To: "Etheridge, Sandra AGF:EX" , > > Message-ID: <022801c4c141$65f66ca0$6639d445@domainnotset.invalid> > Content-Type: text/plain; charset="iso-8859-1" > > Check with your microbiology department. They are probably doing it > already. > > One difference I've found between doing this procedure on smears vs. > fixed/processed tissue - with smears, there is a 5 minute acid-alcohol > decolorization. With fixed/processed/paraffin embedded tissue, reduce this > to 2-3 seconds. The AFB cells walls have been compromised with the > fixative > and alcohols and xylene of the processing. > > Peggy A. Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > ----- Original Message ----- > From: "Etheridge, Sandra AGF:EX" > To: > Sent: Tuesday, November 02, 2004 12:57 PM > Subject: [Histonet] AFB Stain > > > > Hello, all, > > > > Does anyone have a current method for acid fast bacilli using > > auramine-rhodamine? Is this a fluorescent stain? > > > > Thanks in advance. > > > > Sandra Etheridge > > > > BC Ministry of Agriculture, Food & Fisheries > > Animal Health Center, Histology > > Abbotsford, BC > > Canada > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 12 > Date: Wed, 03 Nov 2004 12:33:41 +0200 > From: Marshall > Subject: [Histonet] Merkel cell staining > To: histonet@lists.utsouthwestern.edu > Message-ID: <4188B405.A7723577@cormack.uct.ac.za> > Content-Type: text/plain; charset=us-ascii > > Hi all > I am looking to demonstrate Merkel cells in skin. I can not find any > reference in my textbooks to their demonstration although I have not > used the internet. Also I have tried silver stains for demoing free > nerve endings in skin with not much success. The immunocytochemistry for > neuro filament protein was a bit better but not great. Is there anyway I > can improve on this? > > > > ------------------------------ > > Message: 13 > Date: Wed, 03 Nov 2004 09:41:53 -0500 > From: "Peter Rippstein" > Subject: [Histonet] LR White vs LR Gold > To: > Message-ID: > Content-Type: text/plain; charset="us-ascii" > > Hello Colleagues > > Have any of you had the opportunity to compare EM immunogold labelling > efficiency of LR White vs LR Gold acrylic resins? Protocols would be > helpful. > Many thanks. > > Peter > > Peter Rippstein ART, MLT > University of Ottawa Heart Institute > 40 Ruskin St., Room H453A > Ottawa, Ontario > Canada, K1Y 4W7 > > Tel: (613) 761-5282 > Fax: (613) 761-5281 > email: prippstein@ottawaheart.ca > -------------- next part -------------- > BEGIN:VCARD > VERSION:2.1 > X-GWTYPE:USER > FN:Peter Rippstein > EMAIL;WORK;PREF;NGW:PRippstein@ottawaheart.ca > N:Rippstein;Peter > END:VCARD > > > ------------------------------ > > Message: 14 > Date: Wed, 3 Nov 2004 08:54:40 -0600 > From: Sharon Allen > Subject: [Histonet] RE: Modified Gomori's Trichrome > To: 'Bruce Abaloz' , "Histonet (E-mail)" > > Message-ID: > <304B5A264EC9974E8121B0EA14A9C3936619BA@hsc01mx1.hsc.mb.ca> > Content-Type: text/plain; charset="us-ascii" > > Hi Bruce, > I know the Gomori's Trichrome isn't specific for mitochondria but it > should > stain red if the test is working properly. I have been doing this stain > routinely for 4 different Neuropathologists & am now doing the muscles for > another Neuropathologist who is apparently an expert in the field & > extremely fussy. They all are of the same opinion. Having the mitochondria > stained seems to be imperative for an acceptable stain. Using yet another > stain on the muscle bx's is not an option. This is why I am attempting to > find the key to the "perfect Gomori's Trichrome stain". Using the > celestine > blue before the Mayer's has worked well for staining the mitochondria (I > have done 40 of them), but I have no explanation why. My theory is very > rusty & hoped someone with more knowledge than I could explain the reason > to > me. I think it may have something to do with Celestine Blue staining DNA & > being a oxazine dye. Also chromoptrope 2R being a metachromatic stain. > I would appreciate any help you can give me. > Thanks for your reply, > Sharon > > -----Original Message----- > > This e-mail and/or any documents in this transmission is intended for the > address(s) only and may contain legally privileged or confidential > information. Any unauthorized use, disclosure, distribution, copying or > dissemination is strictly prohibited. If you receive this transmission in > error, please notify the sender immediately and return the original. > > ------------------------------ > > Message: 15 > Date: Wed, 03 Nov 2004 10:32:03 -0500 > From: "Amy Kozer" > Subject: [Histonet] Envision System with Vector Red Background > To: > Message-ID: > Content-Type: text/plain; charset=US-ASCII > > I've tried using Vector Red with Envision system for permanance and have > encountered a lot of background and precipitate. > I've been staining frozen sections of epithelium fixed with acetone. > Has anyone had any experience with this? Remedies? > > Thank you > Amy Kozer > PIADC, ARS, USDA > > > > ------------------------------ > > Message: 16 > Date: Wed, 03 Nov 2004 10:52:46 -0500 > From: "Richard Cartun" > Subject: Re: [Histonet] Merkel cell staining > To: , > Message-ID: > Content-Type: text/plain; charset=US-ASCII > > Try IHC for cytokeratin 20. > > 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 > > >>> Marshall 11/03/04 05:33AM >>> > Hi all > I am looking to demonstrate Merkel cells in skin. I can not find any > reference in my textbooks to their demonstration although I have not > used the internet. Also I have tried silver stains for demoing free > nerve endings in skin with not much success. The immunocytochemistry > for > neuro filament protein was a bit better but not great. Is there anyway > I > can improve on this? > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 17 > Date: Wed, 03 Nov 2004 11:09:31 -0500 > From: Rita Angel > Subject: [Histonet] B & D steamer > To: histonet@lists.utsouthwestern.edu > Message-ID: <5.1.0.14.2.20041103110800.00b24e08@ucmail3.uc.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Thanks to everyone for their input on the type of containers you use in > the > steamer. I'll try the plastic coplin jars!! > > Thanks again, > > Rita Angel HT (ASCP) > > > > > ------------------------------ > > Message: 18 > Date: Wed, 3 Nov 2004 08:52:46 -0800 (PST) > From: Histology SLU > Subject: [Histonet] Polyoma BK virus controls > To: histonet@lists.utsouthwestern.edu > Message-ID: <20041103165246.4952.qmail@web51001.mail.yahoo.com> > Content-Type: text/plain; charset=us-ascii > > Does anyone have BK virus controls that you would be willing to trade for > something we may have that you would need? Any help in locating control > blocks or slides would be greatly appreciated. Thanks. > > Susan > > > --------------------------------- > Do you Yahoo!? > Check out the new Yahoo! Front Page. www.yahoo.com/a > > ------------------------------ > > Message: 19 > Date: Wed, 3 Nov 2004 11:46:59 -0600 > From: "Grant, Debra" > Subject: [Histonet] BMP7 > To: > Message-ID: <1112618780-192831059@pathology.swmed.edu> > Content-Type: text/plain; charset="us-ascii" > > > Has anyone used BMP7 antibody on mouse? If so, could you please send me > all the information, company, catalog #, protocol etc. > > Thanks in advance! > > Debby Grant > Research Technician II > Histology Core Facility > Stowers Institute for Medical Research > 1000 E. 50th Street > Kansas City, MO 64110 > drg@stowers-institute.org > > > > > ------------------------------ > > Message: 20 > Date: Wed, 03 Nov 2004 09:52:01 -0800 > From: "Ant S." > Subject: [Histonet] Thanks for the Teasing > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain > > > Thanks for the great tips on Nerve Teasing. I have quite a few good > ideas to experiment with now. I appreciate the help. This is a very > informative community (and mostly cheerful, too!) > > Thanks, > > Antoinette Swensson > Univeristy of Washington/Harborview Medical Center > Neuropathology > _________________________________________________________________ > > [1]Find the music you love on MSN Music. Start downloading now! > > References > > 1. http://g.msn.com/8HMBENUS/2731??PS=47575 > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 12, Issue 4 > *************************************** > > From lizchlipala <@t> premierhistology.com Wed Nov 3 14:04:05 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] TB block Message-ID: <000501c4c1e0$45969cb0$76d48a80@AMY> Hello everyone I was wondering if there is anyone out there that could spare a block that contains mycobacterium tuberculosis, and be willing to send that to me. I would be willing to pay for the cost of shipping. I'm currently trying to characterize an antibody to TB and the control block of mycobacterium avium that I have will not work for my application. Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From haldana <@t> unimoron.edu.ar Wed Nov 3 16:45:19 2004 From: haldana <@t> unimoron.edu.ar (Hernan Aldana) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Fixation for carbohydrates In-Reply-To: <200410301431171.SM01388@swlx162.swmed.edu> Message-ID: <000001c4c1f6$cc542a40$389559c8@b3w6zzmtb6juvbs> Hello all I need the best fixation method for preserve hyaluronic acid in paraffin sections. Formaldehyde, Bouin, Zencker, B5? Thanks in advance Dr. Hern?n J. Aldana Marcos Facultad de Medicina. Universidad de Mor?n Machado 914. B1708JPD. Buenos Aires. Argentina e-mail alternativo hernanjavier@yahoo.com web: http://hjaldanamarcos.bravepages.com http://www.ht.org.ar From JCarpenter764 <@t> aol.com Wed Nov 3 17:43:22 2004 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] practical exam... Message-ID: has anyone heard about the practical and wether they passed? From beingmary53 <@t> ev1.net Wed Nov 3 17:49:34 2004 From: beingmary53 <@t> ev1.net (maryjohnson) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] exam Message-ID: <003501c4c1ff$c64cf510$b248bbd0@ugly> Hi all I to am wating to hear about the practial exam. From amosbrooks <@t> earthlink.net Wed Nov 3 17:51:51 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Using expired reagents in IHC In-Reply-To: <200411020754.1cp0Z5fq3NZFpK0@mx-a065b01.pas.sa.earthlink.n et> References: <200411020754.1cp0Z5fq3NZFpK0@mx-a065b01.pas.sa.earthlink.net> Message-ID: <6.0.0.22.0.20041103182925.01afbe90@pop.earthlink.net> Hi, We always get new antibodies before the old expire. We only have a few expired ones kicking around. The Big High Muckety-Mucks (somewhat understandably) have determined that the cost of the fines and dealing with battling the inspectors would be worse, in the short term, than the cost of purchasing just enough of the antibodies and not dealing with the extra tech time involved in testing the same antibody monthly. I can see the logic of either side of the coin. It may be easy to overlook a reagent quality gradually decreasing. Repeated testing of the reagent costs time & money too. And the risk of the fines looming over your head is enough to make one hesitate. On the other hand the money saved on keeping antibodies around for decades is appealing. I guess you could say we're erring on the side of caution ... bloody expensive caution. On another note, has anyone noticed that the listed shelf life of these reagents is shrinking lately. I just got one in today that has about 9 months until it expires. It is enough to make one wonder if the antibody companies are pushing for these regulations. I mean, really, where do they come up with these dates anyway? Have a great day, Amos Brooks At 10:54 AM 11/2/2004, you wrote: >Message: 3 >Date: Mon, 01 Nov 2004 14:26:32 -0500 >From: "Richard Cartun" >Subject: [Histonet] Using expired reagents in IHC >To: >Message-ID: >Content-Type: text/plain; charset=US-ASCII > >It has been brought to my attention that the CAP Laboratory checklist >has been revised as of 9/30/04 and now includes a question, "Are all >immunohistochemical reagents used within their indicated expiration >dates?". I have always believed that it is acceptable to use expired >regents (mainly primary antibodies) as long as their reactivity is >acceptable and documented. Obviously, the expiration date that >manufactures use is not an accurate reflection of the reagent's >likelihood of poor performance. Does anyone have information on this >change? Thank you. > >Richard > >Richard W. Cartun, Ph.D. >Director, Immunopathology & Histology >Assistant Director, Anatomic Pathology >Hartford Hospital >80 Seymour Street >Hartford, CT 06102 >(860) 545-1596 >(860) 545-0174 Fax From mrsgbd2001 <@t> yahoo.com Wed Nov 3 18:49:22 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Re: Exam Message-ID: <20041104004922.8210.qmail@web52703.mail.yahoo.com> >From what I've heard, the results for the pratical exam will not be out until late November or early December. Gareth --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com/a From dbpiontek <@t> clinomicsbio.com Wed Nov 3 19:30:29 2004 From: dbpiontek <@t> clinomicsbio.com (Denise Bland-Piontek) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] test Message-ID: <145F33B84793CF488578F86355B32E6912FBA4@mail.clinomicsbio.com> From Dndsomi <@t> aol.com Wed Nov 3 19:29:39 2004 From: Dndsomi <@t> aol.com (Dndsomi@aol.com) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Re: Exam Message-ID: <1a7.2a74b0d2.2ebae003@aol.com> I sent my practical in Sept. 2003 and got my results Jan. 15, 2004. From CrochiereSteve <@t> aol.com Wed Nov 3 20:54:23 2004 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] EGFR Message-ID: <1d8.2f65199e.2ebaf3df@aol.com> Pat, USLabs in California Steven M. Crochiere, HT(ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center Springfield, MA 01104 From Chris.Goodall <@t> bristol.ac.uk Thu Nov 4 07:48:12 2004 From: Chris.Goodall <@t> bristol.ac.uk (Chris.Goodall@bristol.ac.uk) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Timm silver sulphide method for light microscopic localisation of heavy metals. Message-ID: <1099576092.418a331c8b602@webmail.bris.ac.uk> Dear All, I have been asked to try the Timm method on some FFPE samples of human brain. The papers I have read give methods for perfusing rat brain with sodium sulphide solution followed by 3% gluteraldehyde in 0.15M phosphate buffer, and further treatment with sodium sulphide solution, or, post mortem brain samples snap frozen and frozen sectioned. However health and safety rules here do not permit frozen sectioning of unfixed human brain, or the use of gluteradehyde in the embalming room, so I am going to attempt this method on formalin fixed samples.My question is, has anyone experience of this method or any suggestions? It is mentioned that autopsy material left in situ for one or two days post mortem with no fixation may have enough endogenous sulphide ions or sulphide groups in the tissue due to autolytic activity and sulphide treatment may not be necessary, or if it is necessary, does anyone have experience of timing in the sulphide solution and is post fixation necessary after the initial formalin fixation. The paper also mentions that to prevent loss of metallosulphides present in the tissue the temperature of the wax should not exceed 50C which means the use of the dreaded low temperature wax, has anyone used conventional paraffin wax and got away with it? Sorry for the long request,and thank you, Chris Goodall From ajennings <@t> unmc.edu Thu Nov 4 10:03:38 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Histonet 2005 Buttons In-Reply-To: <1099576092.418a331c8b602@webmail.bris.ac.uk> Message-ID: This is it...... I am going to let people continue to submit their designs until next Friday and then I will be presenting the designs to our working group to help me decide which one we will use for the Official 2005 Histonet button. If you have been saving your stuff for last minute.....we are in the 60 second count down for submissions. Please include a space for writing a name, the URL and a place for the company who sponsors the buttons. Thanks a bunch for all the ones we have received it is fun to see the talent and the involvement of the community Anita From mprice26 <@t> juno.com Thu Nov 4 10:16:08 2004 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] RE: Job Openings- Texarkana, TX Message-ID: <20041104.081617.26898.15797@webmail28.nyc.untd.com> Looking for HT (ASCP) Registered Histotech at Pathology Services of Texarkana. Great Location to raise a family. Day shift. Also looking for a temporary histotech for a couple of months. Contact Marsha Price @ mprice26@juno.com. Marsha ________________________________________________________________ Juno Platinum $9.95. Juno SpeedBand $14.95. Sign up for Juno Today at http://www.juno.com! Look for special offers at Best Buy stores. From jkiernan <@t> uwo.ca Thu Nov 4 10:21:19 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Timm silver sulphide method for light microscopiclocalisation of heavy metals. References: <1099576092.418a331c8b602@webmail.bris.ac.uk> Message-ID: <418A56FF.69DDC8C6@uwo.ca> The perfusion of sodium sulphide is necessary to precipitate metal ions (principally zinc), which are otherwise soluble. Relying on autolysis to generate sulphide and precipitate the zinc doesn't make sense because the zinc ions would have diffused away from their original synaptic sites, and the tissue would be decomposing anyway. Timm's method simply won't work (or not in any meaningful way) on formaldehyde-fixed paraffin-embedded specimens. Glutaraldehyde is not the only permissible fixative after perfusing the buffered Na2S. An alternative is to use an alcoholic fixative that has been saturated with H2S gas. It's unlikely that your health & safety regulations will allow this if they won't let you take a little jar of glutaraldehyde into the PM room! The best account of modern Timm staining is probably Danscher,G & Zimmer,J (1978) Histochemistry 55: 27-40. Danscher has also published many more recent papers on histochemical detection of zinc, mercury and other metals with sulphide-silver methods. John Kiernan London, Canada. ___________________________ Chris.Goodall@bristol.ac.uk wrote: > > Dear All, > I have been asked to try the Timm method on some FFPE samples of > human brain. The papers I have read give methods for perfusing rat > brain with sodium sulphide solution followed by 3% gluteraldehyde in > 0.15M phosphate buffer, and further treatment with sodium sulphide > solution, or, post mortem brain samples snap frozen and frozen > sectioned. However health and safety rules here do not permit frozen > sectioning of unfixed human brain, or the use of gluteradehyde in the > embalming room, so I am going to attempt this method on formalin fixed > samples.My question is, has anyone experience of this method or any > suggestions? It is mentioned that autopsy material left in situ for one > or two days post mortem with no fixation may have enough endogenous > sulphide ions or sulphide groups in the tissue due to autolytic > activity and sulphide treatment may not be necessary, or if it is > necessary, does anyone have experience of timing in the sulphide > solution and is post fixation necessary after the initial formalin > fixation. The paper also mentions that to prevent loss of > metallosulphides present in the tissue the temperature of the wax > should not exceed 50C which means the use of the dreaded low > temperature wax, has anyone used conventional paraffin wax and got away > with it? > Sorry for the long request,and thank you, > Chris Goodall > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Nov 4 11:20:44 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] exam In-Reply-To: <003501c4c1ff$c64cf510$b248bbd0@ugly> Message-ID: <000f01c4c292$a0e4f5f0$83020a0a@IHCTech> We just did the grading of over 1100 slide sets for the HT exam last weekend in Chicago, I am sure it will take some extra time this go round to process all those. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of maryjohnson Sent: Wednesday, November 03, 2004 4:50 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] exam Hi all I to am wating to hear about the practial exam. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Nov 4 11:32:45 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] BMP7 In-Reply-To: <1112618780-192831059@pathology.swmed.edu> Message-ID: <001101c4c294$4ba9e3f0$83020a0a@IHCTech> Debra, I have used Santa Cruz goat BMP-7 (L-19) sc-9305 on a lot of rat and human samples but have not tried it on ms. SC lists it as working for rat and h, since it is a goat poly it would be easy to try it on your ms, ask SC if anyone has reported it as working on ms although I don't usually find SC being very helpful for anything not listed being tried. For rats I use it at 1:200 for 1 hr. using LSAB+ detection (I wish someone would come up with a labeled polymer that works with goat antibodies) and pepsin digestion (premade from Phoenix) at 37dc for two changes 5 min. ea. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Grant, Debra Sent: Wednesday, November 03, 2004 10:47 AM To: histonet@pathology.swmed.edu Subject: [Histonet] BMP7 Has anyone used BMP7 antibody on mouse? If so, could you please send me all the information, company, catalog #, protocol etc. Thanks in advance! Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Nov 4 11:48:19 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] bone marrow mush Message-ID: <001701c4c296$79712e40$83020a0a@IHCTech> Folks, I have a question for you. One of my investigators harvested bone marrow by just scrapping it out of the bone cavity, it is not in anything (media, anticoagulant, fixative, etc.) they just stored it at 4dc as a mush in a tube. They want me now to prepare it so that they can see what cells they have. They also want to know if there are any stem cells and what the viability is (by viability I mean they are wondering if they could use it to grow some of the cells in culture). I really don't think I can tell them if they can grow the cells or not. Anyway I was thinking of trying to make a cell block from this material and process into paraffin or maybe even do frozen sections on the pellet. How would you go about making a pellet out of that stuff? Would you put some buffer or media or fixative in the tube and spin it down and then put it in HISTOGEL or wrap in tissue paper and then process? Patsy Ruegg From histobabs <@t> hotmail.com Thu Nov 4 12:05:46 2004 From: histobabs <@t> hotmail.com (B T) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] polyoma virus, SV40 Message-ID: Hi Deb, We are usingSV40 T-Ag (Ab-2) cat# DP02-200ug from oncogene research products. We do the stain on paraffin sections. Barb >From: Browning Deb >To: "'histonet@lists.utsouthwestern.edu'" > >Subject: [Histonet] polyoma virus, SV40 >Date: Mon, 1 Nov 2004 16:16:43 -0500 > >Is anyone out there using an antibody against polyoma virus, SV40; BK; or >JC, against human tissue for demonstrating kidney rejection? If so, could >you share the details, thanks. > >Debra Browning, ART >Technical Specialist, Immunohistochemistry >Anatomic Pathology >Hamilton Health Sciences >phone: (905) 527-4322 ext 46131 >e-mail: browning@hhsc.ca >fax: (905) 524-2681 > > >This information is directed in confidence solely to the person named above >and may not otherwise be distributed, copied or disclosed. Therefore, this >information should be considered strictly confidential. If you have >received this email in error, please notify the sender immediately via a >return email for further direction. Thank you for your assistance. > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Thu Nov 4 12:09:44 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Timm silver sulphide method... References: <418A2EEF.21922.1002BFE@localhost> Message-ID: <418A7068.A6A5AD89@uwo.ca> For insoluble metal-containing deposits, yes. But the zinc in the brain is in solution in certain synaptic vesicles. The zinc in leukocytes and Paneth cells stays in place after alcohol fixation and can be stained with dithizone, which is a less sensitive method than Timm's. John Kiernan London, Canada. --------------------------------------------- Greg Dobbin wrote: > > Hi Chris and John, > Chris, you did not say which metals you hoped to stain. > John you state: > "Timm's method simply won't work (or not > in any meaningful way) on formaldehyde-fixed > paraffin-embedded specimens." > While I do not have any experience with the method described by Chris using perfusion, I have on many occasions successfully stained FFPE sections with a modified Timm's Stain for heavy metals in general (ie not distinguishing one metal from another) and for copper specifically. I have offered the method and reference several times over the years here on the Histonet. > > The reference again is: > ?Diagnostic Histochemistry?; Frederick T. Zugibe > Copyright 1970 by The C.V. Mosby Company > (Standard Book Number [ISBN?] 8016-5702-4) > > If either of you wish to receive a copy of the method with a few of my own observations, I would be happy to send it along in the form of an attachment. Just indicate whether you want it in a WordPerfect or Microsoft Word format. > Cheers! > Greg > > Date sent: Thu, 04 Nov 2004 11:21:19 -0500 > From: John Kiernan > To: Chris.Goodall@bristol.ac.uk > Copies to: histonet@lists.utsouthwestern.edu > Send reply to: jkiernan@uwo.ca > related fields > > > Subject: Re: [Histonet] Timm silver sulphide method for light > microscopiclocalisation of heavy metals. > > > The perfusion of sodium sulphide is necessary > > to precipitate metal ions (principally zinc), > > which are otherwise soluble. Relying on autolysis > > to generate sulphide and precipitate the zinc > > doesn't make sense because the zinc ions would > > have diffused away from their original synaptic > > sites, and the tissue would be decomposing > > anyway. Timm's method simply won't work (or not > > in any meaningful way) on formaldehyde-fixed > > paraffin-embedded specimens. > > > > Glutaraldehyde is not the only permissible > > fixative after perfusing the buffered Na2S. > > An alternative is to use an alcoholic fixative > > that has been saturated with H2S gas. It's > > unlikely that your health & safety regulations > > will allow this if they won't let you take a > > little jar of glutaraldehyde into the PM room! > > > > The best account of modern Timm staining is > > probably Danscher,G & Zimmer,J (1978) > > Histochemistry 55: 27-40. Danscher has also > > published many more recent papers on > > histochemical detection of zinc, mercury and > > other metals with sulphide-silver methods. > > > > John Kiernan > > London, Canada. > > ___________________________ > > Chris.Goodall@bristol.ac.uk wrote: > > > > > > Dear All, > > > I have been asked to try the Timm method on some FFPE samples of > > > human brain. The papers I have read give methods for perfusing rat > > > brain with sodium sulphide solution followed by 3% gluteraldehyde in > > > 0.15M phosphate buffer, and further treatment with sodium sulphide > > > solution, or, post mortem brain samples snap frozen and frozen > > > sectioned. However health and safety rules here do not permit frozen > > > sectioning of unfixed human brain, or the use of gluteradehyde in the > > > embalming room, so I am going to attempt this method on formalin fixed > > > samples.My question is, has anyone experience of this method or any > > > suggestions? It is mentioned that autopsy material left in situ for one > > > or two days post mortem with no fixation may have enough endogenous > > > sulphide ions or sulphide groups in the tissue due to autolytic > > > activity and sulphide treatment may not be necessary, or if it is > > > necessary, does anyone have experience of timing in the sulphide > > > solution and is post fixation necessary after the initial formalin > > > fixation. The paper also mentions that to prevent loss of > > > metallosulphides present in the tissue the temperature of the wax > > > should not exceed 50C which means the use of the dreaded low > > > temperature wax, has anyone used conventional paraffin wax and got away > > > with it? > > > Sorry for the long request,and thank you, > > > Chris Goodall > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Research Technologist, Pathology Lab, Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From Rachael_Emerson <@t> URMC.Rochester.edu Thu Nov 4 12:14:07 2004 From: Rachael_Emerson <@t> URMC.Rochester.edu (Emerson, Rachael) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] RE: bonw marrow mush Message-ID: Hello, I do not think that bone marrow frozen at 4dc straight from dissection is going to have any viability. I would refer to Stem Cell Technologies website. The have kits and reagents, as well as, great manuals for this kind of stuff. Rachael Emerson 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: histonet-request@lists.utsouthwestern.edu > Reply To: histonet@lists.utsouthwestern.edu > Sent: Thursday, November 4, 2004 1:04 PM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet Digest, Vol 12, Issue 5 > > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. CD41, GPV, GP1bBeta (Emerson, Rachael) > 2. TB block (Elizabeth Chlipala) > 3. Fixation for carbohydrates (Hernan Aldana) > 4. practical exam... (JCarpenter764@aol.com) > 5. exam (maryjohnson) > 6. Using expired reagents in IHC (Amos Brooks) > 7. Re: Exam (Gareth Davis) > 8. test (Denise Bland-Piontek) > 9. Re: Re: Exam (Dndsomi@aol.com) > 10. EGFR (CrochiereSteve@aol.com) > 11. Timm silver sulphide method for light microscopic > localisation of heavy metals. (Chris.Goodall@bristol.ac.uk) > 12. Histonet 2005 Buttons (ajennings@unmc.edu) > 13. RE: Job Openings- Texarkana, TX (mprice26@juno.com) > 14. Re: Timm silver sulphide method for light > microscopiclocalisation of heavy metals. (John Kiernan) > 15. RE: exam (Patsy Ruegg) > 16. RE: BMP7 (Patsy Ruegg) > 17. bone marrow mush (Patsy Ruegg) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Wed, 3 Nov 2004 14:00:53 -0500 > From: "Emerson, Rachael" > Subject: [Histonet] CD41, GPV, GP1bBeta > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > > Content-Type: text/plain; charset="iso-8859-1" > > Hi. Does anyone have any experience working with CD41, GPV, or GP1bBeta > antibodies? > I could really use some help. > > Thanks a lot!! > Rachael Emerson > > 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: histonet-request@lists.utsouthwestern.edu > > Reply To: histonet@lists.utsouthwestern.edu > > Sent: Wednesday, November 3, 2004 1:03 PM > > To: histonet@lists.utsouthwestern.edu > > Subject: Histonet Digest, Vol 12, Issue 4 > > > > Send Histonet mailing list submissions to > > histonet@lists.utsouthwestern.edu > > > > To subscribe or unsubscribe via the World Wide Web, visit > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > or, via email, send a message with subject or body 'help' to > > histonet-request@lists.utsouthwestern.edu > > > > You can reach the person managing the list at > > histonet-owner@lists.utsouthwestern.edu > > > > When replying, please edit your Subject line so it is more specific > > than "Re: Contents of Histonet digest..." > > > > > > Today's Topics: > > > > 1. Alcian Blue and steamer containers (Paula Pierce) > > 2. Re: Sta-On (Jennifer Sipes) > > 3. Epidermal growth factor (Ford, Rhonda) > > 4. Job posting - Dallas (Patterson, Pat) > > 5. RE: thanks and aquamount (Patsy Ruegg) > > 6. RE: AFB Stain (Joe Nocito) > > 7. Re: History of microtechnique (John Kiernan) > > 8. Re: GMA sectioning (Caroline Stott) > > 9. Re: sample preparation/analysis to observe gas bubbles > > (Wayne Kreider) > > 10. Re: Polyoma Virus (tdobersztyn@chmca.org) > > 11. Re: AFB Stain (lpwenk@sbcglobal.net) > > 12. Merkel cell staining (Marshall) > > 13. LR White vs LR Gold (Peter Rippstein) > > 14. RE: Modified Gomori's Trichrome (Sharon Allen) > > 15. Envision System with Vector Red Background (Amy Kozer) > > 16. Re: Merkel cell staining (Richard Cartun) > > 17. B & D steamer (Rita Angel) > > 18. Polyoma BK virus controls (Histology SLU) > > 19. BMP7 (Grant, Debra) > > 20. Thanks for the Teasing (Ant S.) > > > > > > ---------------------------------------------------------------------- > > > > Message: 1 > > Date: Tue, 2 Nov 2004 10:06:53 -0800 (PST) > > From: Paula Pierce > > Subject: [Histonet] Alcian Blue and steamer containers > > To: histonet@lists.utsouthwestern.edu > > Message-ID: <20041102180653.38389.qmail@web50308.mail.yahoo.com> > > Content-Type: text/plain; charset=us-ascii > > > > Hello Andrew, > > > > yes I have already voted today :) > > > > You can buy a smaller amount of alcian blue at > > http://www.anatechltdusa.com/Catalog/catalogspecialstains.html > > > > I use plastic coplin jars in my B&D steamer for small quantities of > slides > > and the larger tissue-tek when I have more. > > > > > > 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, 2 Nov 2004 10:10:48 -0800 (PST) > > From: Jennifer Sipes > > Subject: [Histonet] Re: Sta-On > > To: histonet@lists.utsouthwestern.edu > > Message-ID: <20041102181048.98641.qmail@web60604.mail.yahoo.com> > > Content-Type: text/plain; charset=us-ascii > > > > I purchased it through a company called Surgi-Path. They're the ones > that > > make it. > > > > > > 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!? > > Check out the new Yahoo! Front Page. www.yahoo.com/a > > > > ------------------------------ > > > > Message: 3 > > Date: Tue, 2 Nov 2004 13:11:38 -0500 > > From: "Ford, Rhonda" > > Subject: [Histonet] Epidermal growth factor > > To: > > Message-ID: > > <684DF8CDE1FB6A428499DA65D753D12201F61789@JUPITER.HCMH.ORG> > > Content-Type: text/plain; charset="us-ascii" > > > > Does anyone know any institution(s) who perform this testing? > > > > > > ------------------------------ > > > > Message: 4 > > Date: Tue, 2 Nov 2004 12:26:35 -0600 > > From: "Patterson, Pat" > > Subject: [Histonet] Job posting - Dallas > > To: "'histonet@lists.utsouthwestern.edu'" > > > > Message-ID: <293C7C19EFF7D611AE1A0002A53F81140CB43B0D@omega.mhd.com> > > Content-Type: text/plain; charset="iso-8859-1" > > > > > > Named one of the "Best Places to work 2004" by the Dallas Business > > Journal. > > > > Our dedicated team is very outspoken in their appreciation of the > > excellent > > clinical environment we foster and the ongoing commitment Methodist has > > made > > to patient satisfaction. Our dual focus on out patients and our > employees > > is working - 2002 Methodist Health System earned the Gold Seal of > > Approval > > form the Joint Commission on Accreditation of Healthcare Organizations > and > > more than 28 percent of our new hires come from employee referral! If > > excellent patient care is one of your priorities for career > satisfaction, > > then you belong at Methodist. > > > > > > > > Join our Histology team at Methodist Dallas Medical Center. We're > seeking > > a > > full-time Histology Technician on the day shift in our busy Anatomic > > Pathology Section. Job responsibilities include processing, embedding, > > sectioning, routine and special staining, assisting in gross room, > frozen > > sectioning and computer entry into the MediTech lab information system. > > Experience with microwave technology would be an added plus. Requires > > HT/HTL > > ASCP registry or eligible. Minimum 2-year experience desired. > > > > Contact: Pat Patterson, Manager > > Phone: (214) 947-3538 > > Fax: (214) 947-3524 > > email: PatPatterson@mhd.com > > > > > > *********************************************************************** > > > > This electronic transmission contains information from Methodist Health > > System and should be considered confidential and privileged. The > > information contained in the above messages is intended only for the > > use of the individual(s) and entity(ies) named above. If you are not > > the intended recipient, be aware that any disclosure, copying, > > distribution, or use of this information is prohibited. If you receive > > this transmission in error, please notify the sender immediately by > > return e-mail. Methodist Health System, its subsidiaries and > > affiliates hereby claim all applicable privileges related to the > > transmission of this communication. > > > > > > > > ------------------------------ > > > > Message: 5 > > Date: Tue, 2 Nov 2004 11:33:43 -0700 > > From: "Patsy Ruegg" > > Subject: RE: [Histonet] thanks and aquamount > > To: "'Sharon Cooperman'" , > > > > Message-ID: <001501c4c10a$7b4e4fa0$83020a0a@IHCTech> > > Content-Type: text/plain; charset="us-ascii" > > > > Sharon, > > Aquamount will not harden, you should seal with clear nail polish to > > keep it permanently. > > Patsy > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon > > Cooperman > > Sent: Thursday, October 28, 2004 12:16 PM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] thanks and aquamount > > > > > > Dear Histonetters, > > > > Thanks to everyone for the great advice on bone decal and oil red o > > stain. I have one last question related to the oil red o stain - I > > just bought a bottle of aquamount to use when mounting the oil red o > > stained slides, but it didn't come with instructions (on the bottle > > it refers to instructions). Are there any special instructions for > > aquamount? Do I need to seal the slides with nail polish or does > > aquamount harden? Or, does anyone know how I can contact Lerner to > > get a copy of the instructions (I bought it from Fisher - Lerner > > doesn't answer the phone number I have for them). > > > > Thanks, > > Sharon > > -- > > Sharon Cooperman > > NIH, NICHD, CBMB 301.435-8417 > > Building 18T, room 101 301.402-0078 fax > > Bethesda, MD 20892 > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > > ------------------------------ > > > > Message: 6 > > Date: Tue, 2 Nov 2004 12:58:15 -0600 > > From: "Joe Nocito" > > Subject: RE: [Histonet] AFB Stain > > To: "Etheridge, Sandra AGF:EX" , > > > > Message-ID: > > Content-Type: text/plain; charset="iso-8859-1" > > > > Sandra, > > attached is our procedure for Truant's AFB. We purchase the staining kit > > from Infolab. > > > > > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of > > Etheridge, Sandra AGF:EX > > Sent: Tuesday, November 02, 2004 11:57 AM > > To: (histonet@lists.utsouthwestern.edu) > > Subject: [Histonet] AFB Stain > > > > > > Hello, all, > > > > Does anyone have a current method for acid fast bacilli using > > auramine-rhodamine? Is this a fluorescent stain? > > > > Thanks in advance. > > > > Sandra Etheridge > > > > BC Ministry of Agriculture, Food & Fisheries > > Animal Health Center, Histology > > Abbotsford, BC > > Canada > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > > > Message: 7 > > Date: Tue, 02 Nov 2004 14:19:49 -0500 > > From: John Kiernan > > Subject: Re: [Histonet] History of microtechnique > > To: MTitford@aol.com > > Cc: Histonet@lists.utsouthwestern.edu > > Message-ID: <4187DDD5.6F8C39E1@uwo.ca> > > Content-Type: text/plain; charset=us-ascii > > > > There is also a chapter on the History of Staining > > by F. Kasten in the 10th edition of "Conn's > > Biological Stains." This book (2002) is > > still in print. > > ------------------------------- > > John A. Kiernan > > Department of Anatomy and Cell Biology > > The University of Western Ontario > > London, Canada N6A 5C1 > > _______________________________ > > MTitford@aol.com wrote: > > > > > > David Muskett asks about where to find information about the history > of > > microtechnique. > > > A good place to start is with "A history of microtechnique" by Brian > > Bracegirdle. The second edition is still available here in the States > from > > Science Heritage Limited, Lincolnwood, IL > > > a second good book is "History of staining" by George Clark and > > Frederick Kasten. 3ed edition, Williams & Wilkins, Baltimore & London. > The > > second book is probably out of print but available on interlibrary loan. > > > There are many other sources but you can get information overload in > > this area! > > > > > > Mike Titford > > > USA Pathology > > > Mobile AL > > > > > > > > ------------------------------ > > > > Message: 8 > > Date: Wed, 03 Nov 2004 09:49:23 +1300 > > From: Caroline Stott > > Subject: [Histonet] Re: GMA sectioning > > To: Histonet@lists.utsouthwestern.edu > > Message-ID: <5.2.1.1.0.20041103094227.02076a10@anatomy.otago.ac.nz> > > Content-Type: text/plain; charset="us-ascii"; format=flowed > > > > Hi Ben, > > We have quite regularly cut GMA at 10, 20, 30 and 40 microns. We use a > > cotton bud to apply a little water to the face of the block, just gently > > > rubbing it in. Also place some water on the glass knife as the section > > will come off easier. Then take a section and push it to the bottom of > > the > > water bowl/dish so it will flatten out. Then slide the sections onto > the > > slide. Heres the tricky part.... We put a piece of filter paper on the > > > bench then the slide on top, then another piece of wet filter paper on > top > > > > of the slide (and sections) and use a roller (like a paint roller) > > applying a little pressure while rolling around 20-30 times over the > slide > > > > and finally place on a hotplate (not flat) that is around 60 degrees > > C. Haven't had a problem. :) > > Hope that makes sense. > > > > Caroline > > > > Caroline Stott > > > > Histology Service Unit > > University of Otago > > PO Box 913 > > Dunedin, New Zealand > > Ph (03) 479 7152 > > Fax (03) 479 7136 > > > > ------------------------------ > > > > Message: 9 > > Date: Tue, 02 Nov 2004 12:54:20 -0800 > > From: Wayne Kreider > > Subject: Re: [Histonet] sample preparation/analysis to observe gas > > bubbles > > To: Philip Oshel > > Cc: histonet@lists.utsouthwestern.edu > > Message-ID: <4187F3FC.3060400@u.washington.edu> > > Content-Type: text/plain; charset=ISO-8859-1; format=flowed > > > > Phil, > > Thanks again... all very helpful. I think we can now plan how we'll > > have the best chance at 'catching' transient bubbles. I'm not sure > > we'll have our techniques together for sampling and freezing the tissue > > in the very near term (this week), but I am hopeful we can do/try this. > > > By the way, you mentioned that frozen samples are not useful for EM... > > how so? Perhaps due to subsequent fixing procedures? I'm relatively > > unfamiliar with EM although TEM had been suggested for this work. > > Wayne > > > > Philip Oshel wrote: > > > > > Wayne, > > > > > > You do have a problem ... > > > 2 X 2 X 2 mm is too big for any proper freezing method. Even > > > high-pressure freezing, which can in ideal conditions give the > > > greatest depth of good freezing, only does at best 500 microns. The > > > other methods do 200 microns or less. Plunging into LN2-cooled > > > cryogens will do a few 10s of microns, no more. They're fine for light > > > > microscopy (assuming migration of molecules from holed membranes or > > > desiccation isn't an issue), but not EM. > > > The major source of freezing damage isn't really mechanical > > > ice-crystal growth and damage, but dehydration of cells from crystal > > > growth. > > > Plunging is literally dropping the sample into the cryogen, but not > > > really. By then I mean you *rapidly* plunge it in, not just let it > > > fall. By hand, as fast as possible. This is important in order to get > > > through the layer of cold nitrogen above the cryogen (whatever it is). > > > > Some commercial plungers are designed to drop the sample into the > > > cryogen from enough height to pass through the cold gas layer quickly. > > > > These are used for electrophysiology -- stimulate the sample then > > > drop, or stimulate during the drop, and therefore the sample is frozen > > > > X msec after stimulation. > > > I can see doing this with ultrasonication, depending on the size, etc. > > > > of the sonciation apparatus. > > > Cryoprotectants. Um. I don't think you can use these. The tissue has > > > to soak in the cryoprotectant -- often moving up a gradient -- in > > > order to get the cryoprotectant completely infiltrated into the > > > sample. This takes an hour or more -- maybe several hours? You'd > > > either have to assume nothing happens with the bubbles in the tissue > > > while sitting around infiltrating with cryoprotectant, or you'd have > > > to get the cryoprotectant into the tissue first, then sonicate and > > > pretend the cryoprotectant doesn't change the tissue properties and > > > therefore resulting bubbles. > > > I wouldn't believe either of those choices. > > > I think you'll need to do the experiment, then freeze, and use pieces > > > of tissue small enough to freeze properly. But! It's not entirely bad > > > -- only one dimension has to be =<200 microns -- 100 microns, really. > > > The other dimensions could be 2 mm. > > > So: 2 mm X 2 mm X 100 microns (even thinner would be better) should > > > work. Just hold the tissue by a corner, so that the cryogen has > > > maximal access to both faces of the thin side. And move the tissue > > > once in the slush LN2, don't hold it in one place -- keep fresh > > > cryogen contacting the sample. > > > Let's see -- oh. Insulate the container (and vacuum desiccator) as > > > best as possible, and work quickly, since you'll only have a few > > > minutes to freeze before the slush LN2 warms up to regular > > > almost-boiling LN2. > > > (I think Bal-Tec makes an apparatus for making slush LN2, and maybe > > > freezing with it, but you don't really need it, it just maybe makes > > > things easier.) > > > > > > Phil > > > > > > Phil, > > > Thanks for the input. Unfortunately, I haven't had much luck looking > > > in the archive. > > > The treatment is actually acoustic (high intensity ultrasound), so no > > > electrodes are involved. Our treatment volume that we'd ideally like > > > to freeze is about 2mm x 2mm x 5mm. However, from the little I've > > > read about plunging, a guideline for the maximum sample thickness is > > > 0.2mm. Using a slush as you describe would presumably help some, but > > > I suspect 2mm is still too thick. > > > If we are able to obtain a suitably thin sample, is plunging literally > > > > just dropping the raw tissue into the cryogen? Or are there some > > > additional steps that should be taken in handling the sample? > > > Moreover, if we do have a larger sample, reference books suggest that > > > it's necessary to use a cryo-protectant. Is a cryo-protectant likely > > > to distort the tissue, in particular any bubble-type structures? > > > Also, what time delays are typically associated with using a > > > protectant? Any suggestions about which cryo-protectant might be best > > > > suited for this application? Thanks again... I'd appreciate any > > > thoughts you might have. > > > Wayne > > > > > > Philip Oshel wrote: > > > > > > Cryofixation is the only way I know to preserve this kind of > > > structure. There have been discussions about this in the past, so you > > > might want to poke around in the Histonet archives. > > > What kind of treatment? Is it something that you stimulate with say an > > > > electrode? > > > If so, there are plunge freezers designed to allow you to stimulate > > > the sample while it is falling into the liquid nitrogen. > > > Rapidity of freezing is essential to insure vitrification of the > > > water, instead of crystal formation. Although some folks maintain that > > > > the water doesn't vitrify, but instead forms micro-crystallites small > > > enough to have no effect on structure. Others state that evanescent > > > microspherules are formed, sort of neither fish nor fowl -- not > > > crystalline and not glass. Either way, the frozen samples have to be > > > maintained below the recrystallization temperature (around -80deg C or > > > > so) until after dehydraton or embedding to prevent crystal growth and > > > negating all the rapid freezing goodness. > > > But, the 4 basic methods are high-pressure freezing, propane-jet > > > freezing, slam-freezing, and plunging into cryogen. The first three > > > will likely distort the air bubbles and tissue around them, however. > > > Plunging works well, as long as the plunge is made very rapidly, and > > > no time is spent hanging about in the cold -- well below freezing -- > > > nitrogen atmosphere above the cryogen. > > > Many people plunge into some organic fluid like ethane which is held > > > in a container in liquid nitrogen. This works, but it has a few > > > issues: the cryogen is usually warmer than the LN2, so the freezing is > > > > not as rapid as it needs to be, and such organic cryogens are > > > flammable or explosive when they warm up. Especially since, being near > > > > LN2 temperatures, they're enriching themselves in oxygen from the air > > > (liquid air is warmer than liquid nitrogen). They can be handled > > > safely, but this does require some thought. > > > A better way is to plunge into slush nitrogen -- LN2 near the freezing > > > > point, instead of near the boiling point. This is about 14 deg C > > > colder than LN2, so freezes samples faster, and so gives a greater > > > depth of good freezing. It's also nitrogen, so there's no flammable > > > gas to deal with (nor bureaucrats upset about flammable gases in your > > > lab). It's easy to produce: just put a beaker full in a small to > > > medium size vacuum desiccator, and pull a vacuum with a high capacity > > > pump, like a dual-stage rotary pump. One used to rough out a large EM > > > column usually works. > > > > > > Phil > > > > > > We're exploring a treatment in which small short-lived (perhaps on the > > > > order of seconds) gas bubbles may be generated in tissue. To this > > > end, we're interested in learning the 'instantaneous' state of the > > > tissue immediately after treatment with regard to the presence of any > > > bubbles. Currently, our experiments utilize rabbit muscle. Are there > > > any known histological sample preparation/analysis procedures that > > > have been used to preserve/observe resident gas bubbles? We have > > > considered a flash freezing followed by staining and/or electron > > > microscopy. However, our expertise is not in histology, so we'd very > > > much appreciate any suggestions and/or details regarding possible > > > sample preparation. Thanks. > > > Wayne > > > -- > > > > > > /**********************/ > > > > /Wayne Kreider/ > > > > /University of Washington/ > > > > /Applied Physics Lab / CIMU/ > > > > /106 Old Fisheries/ > > > > /206-543-1324/ > > > > > > > > ------------------------------ > > > > Message: 10 > > Date: Tue, 2 Nov 2004 12:55:07 -0500 > > From: tdobersztyn@chmca.org > > Subject: [Histonet] Re: Polyoma Virus > > To: histonet@lists.utsouthwestern.edu > > Message-ID: > > Content-Type: text/plain; charset=US-ASCII > > > > > > > > > > > > > > > > > > Debra- > > BK, Polyoma virus, can be demonstrated beautifully with Electron > > Microscopy. > > That is, if you have access to an EM lab. > > I am frequently processing renal core biopsies to r/o BK. > > If you would like assistance- > > let me know. > > > > > > > > Date: Mon, 1 Nov 2004 16:16:43 -0500 > > From: Browning Deb > > Subject: [Histonet] polyoma virus, SV40 > > To: "'histonet@lists.utsouthwestern.edu'" > > > > Message-ID: > > > > <3AADFB88753AD31189C100902786B91C0E27851F@hch_nt_exchange.hhsc.ca> > > Content-Type: text/plain; charset="iso-8859-1" > > > > Is anyone out there using an antibody against polyoma virus, SV40; BK; > or > > JC, against human tissue for demonstrating kidney rejection? If so, > could > > you share the details, thanks. > > > > Debra Browning, ART > > Technical Specialist, Immunohistochemistry > > Anatomic Pathology > > Hamilton Health Sciences > > phone: (905) 527-4322 ext 46131 > > e-mail: browning@hhs > > > > > > > > > > > > Theresa R Dobersztyn HT ASCP > > Electron Microscopy Laboratory > > Department of Pathology > > > > Akron Children's Hospital > > 1 Perkins Square > > Akron, Ohio 44308 > > > > 330-543-8279 > > > > > > > > > > ------------------------------ > > > > Message: 11 > > Date: Tue, 2 Nov 2004 20:06:49 -0500 > > From: > > Subject: Re: [Histonet] AFB Stain > > To: "Etheridge, Sandra AGF:EX" , > > > > Message-ID: <022801c4c141$65f66ca0$6639d445@domainnotset.invalid> > > Content-Type: text/plain; charset="iso-8859-1" > > > > Check with your microbiology department. They are probably doing it > > already. > > > > One difference I've found between doing this procedure on smears vs. > > fixed/processed tissue - with smears, there is a 5 minute acid-alcohol > > decolorization. With fixed/processed/paraffin embedded tissue, reduce > this > > to 2-3 seconds. The AFB cells walls have been compromised with the > > fixative > > and alcohols and xylene of the processing. > > > > Peggy A. Wenk, HTL(ASCP)SLS > > William Beaumont Hospital > > Royal Oak, MI 48073 > > > > ----- Original Message ----- > > From: "Etheridge, Sandra AGF:EX" > > To: > > Sent: Tuesday, November 02, 2004 12:57 PM > > Subject: [Histonet] AFB Stain > > > > > > > Hello, all, > > > > > > Does anyone have a current method for acid fast bacilli using > > > auramine-rhodamine? Is this a fluorescent stain? > > > > > > Thanks in advance. > > > > > > Sandra Etheridge > > > > > > BC Ministry of Agriculture, Food & Fisheries > > > Animal Health Center, Histology > > > Abbotsford, BC > > > Canada > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > > ------------------------------ > > > > Message: 12 > > Date: Wed, 03 Nov 2004 12:33:41 +0200 > > From: Marshall > > Subject: [Histonet] Merkel cell staining > > To: histonet@lists.utsouthwestern.edu > > Message-ID: <4188B405.A7723577@cormack.uct.ac.za> > > Content-Type: text/plain; charset=us-ascii > > > > Hi all > > I am looking to demonstrate Merkel cells in skin. I can not find any > > reference in my textbooks to their demonstration although I have not > > used the internet. Also I have tried silver stains for demoing free > > nerve endings in skin with not much success. The immunocytochemistry for > > neuro filament protein was a bit better but not great. Is there anyway I > > can improve on this? > > > > > > > > ------------------------------ > > > > Message: 13 > > Date: Wed, 03 Nov 2004 09:41:53 -0500 > > From: "Peter Rippstein" > > Subject: [Histonet] LR White vs LR Gold > > To: > > Message-ID: > > Content-Type: text/plain; charset="us-ascii" > > > > Hello Colleagues > > > > Have any of you had the opportunity to compare EM immunogold labelling > > efficiency of LR White vs LR Gold acrylic resins? Protocols would be > > helpful. > > Many thanks. > > > > Peter > > > > Peter Rippstein ART, MLT > > University of Ottawa Heart Institute > > 40 Ruskin St., Room H453A > > Ottawa, Ontario > > Canada, K1Y 4W7 > > > > Tel: (613) 761-5282 > > Fax: (613) 761-5281 > > email: prippstein@ottawaheart.ca > > -------------- next part -------------- > > BEGIN:VCARD > > VERSION:2.1 > > X-GWTYPE:USER > > FN:Peter Rippstein > > EMAIL;WORK;PREF;NGW:PRippstein@ottawaheart.ca > > N:Rippstein;Peter > > END:VCARD > > > > > > ------------------------------ > > > > Message: 14 > > Date: Wed, 3 Nov 2004 08:54:40 -0600 > > From: Sharon Allen > > Subject: [Histonet] RE: Modified Gomori's Trichrome > > To: 'Bruce Abaloz' , "Histonet (E-mail)" > > > > Message-ID: > > <304B5A264EC9974E8121B0EA14A9C3936619BA@hsc01mx1.hsc.mb.ca> > > Content-Type: text/plain; charset="us-ascii" > > > > Hi Bruce, > > I know the Gomori's Trichrome isn't specific for mitochondria but it > > should > > stain red if the test is working properly. I have been doing this stain > > routinely for 4 different Neuropathologists & am now doing the muscles > for > > another Neuropathologist who is apparently an expert in the field & > > extremely fussy. They all are of the same opinion. Having the > mitochondria > > stained seems to be imperative for an acceptable stain. Using yet > another > > stain on the muscle bx's is not an option. This is why I am attempting > to > > find the key to the "perfect Gomori's Trichrome stain". Using the > > celestine > > blue before the Mayer's has worked well for staining the mitochondria (I > > have done 40 of them), but I have no explanation why. My theory is very > > rusty & hoped someone with more knowledge than I could explain the > reason > > to > > me. I think it may have something to do with Celestine Blue staining DNA > & > > being a oxazine dye. Also chromoptrope 2R being a metachromatic stain. > > I would appreciate any help you can give me. > > Thanks for your reply, > > Sharon > > > > -----Original Message----- > > > > This e-mail and/or any documents in this transmission is intended for > the > > address(s) only and may contain legally privileged or confidential > > information. Any unauthorized use, disclosure, distribution, copying or > > dissemination is strictly prohibited. If you receive this transmission > in > > error, please notify the sender immediately and return the original. > > > > ------------------------------ > > > > Message: 15 > > Date: Wed, 03 Nov 2004 10:32:03 -0500 > > From: "Amy Kozer" > > Subject: [Histonet] Envision System with Vector Red Background > > To: > > Message-ID: > > Content-Type: text/plain; charset=US-ASCII > > > > I've tried using Vector Red with Envision system for permanance and have > > encountered a lot of background and precipitate. > > I've been staining frozen sections of epithelium fixed with acetone. > > Has anyone had any experience with this? Remedies? > > > > Thank you > > Amy Kozer > > PIADC, ARS, USDA > > > > > > > > ------------------------------ > > > > Message: 16 > > Date: Wed, 03 Nov 2004 10:52:46 -0500 > > From: "Richard Cartun" > > Subject: Re: [Histonet] Merkel cell staining > > To: , > > Message-ID: > > Content-Type: text/plain; charset=US-ASCII > > > > Try IHC for cytokeratin 20. > > > > 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 > > > > >>> Marshall 11/03/04 05:33AM >>> > > Hi all > > I am looking to demonstrate Merkel cells in skin. I can not find any > > reference in my textbooks to their demonstration although I have not > > used the internet. Also I have tried silver stains for demoing free > > nerve endings in skin with not much success. The immunocytochemistry > > for > > neuro filament protein was a bit better but not great. Is there anyway > > I > > can improve on this? > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > ------------------------------ > > > > Message: 17 > > Date: Wed, 03 Nov 2004 11:09:31 -0500 > > From: Rita Angel > > Subject: [Histonet] B & D steamer > > To: histonet@lists.utsouthwestern.edu > > Message-ID: <5.1.0.14.2.20041103110800.00b24e08@ucmail3.uc.edu> > > Content-Type: text/plain; charset="us-ascii"; format=flowed > > > > Thanks to everyone for their input on the type of containers you use in > > the > > steamer. I'll try the plastic coplin jars!! > > > > Thanks again, > > > > Rita Angel HT (ASCP) > > > > > > > > > > ------------------------------ > > > > Message: 18 > > Date: Wed, 3 Nov 2004 08:52:46 -0800 (PST) > > From: Histology SLU > > Subject: [Histonet] Polyoma BK virus controls > > To: histonet@lists.utsouthwestern.edu > > Message-ID: <20041103165246.4952.qmail@web51001.mail.yahoo.com> > > Content-Type: text/plain; charset=us-ascii > > > > Does anyone have BK virus controls that you would be willing to trade > for > > something we may have that you would need? Any help in locating > control > > blocks or slides would be greatly appreciated. Thanks. > > > > Susan > > > > > > --------------------------------- > > Do you Yahoo!? > > Check out the new Yahoo! Front Page. www.yahoo.com/a > > > > ------------------------------ > > > > Message: 19 > > Date: Wed, 3 Nov 2004 11:46:59 -0600 > > From: "Grant, Debra" > > Subject: [Histonet] BMP7 > > To: > > Message-ID: <1112618780-192831059@pathology.swmed.edu> > > Content-Type: text/plain; charset="us-ascii" > > > > > > Has anyone used BMP7 antibody on mouse? If so, could you please send me > > all the information, company, catalog #, protocol etc. > > > > Thanks in advance! > > > > Debby Grant > > Research Technician II > > Histology Core Facility > > Stowers Institute for Medical Research > > 1000 E. 50th Street > > Kansas City, MO 64110 > > drg@stowers-institute.org > > > > > > > > > > ------------------------------ > > > > Message: 20 > > Date: Wed, 03 Nov 2004 09:52:01 -0800 > > From: "Ant S." > > Subject: [Histonet] Thanks for the Teasing > > To: histonet@lists.utsouthwestern.edu > > Message-ID: > > Content-Type: text/plain > > > > > > Thanks for the great tips on Nerve Teasing. I have quite a few > good > > ideas to experiment with now. I appreciate the help. This is a > very > > informative community (and mostly cheerful, too!) > > > > Thanks, > > > > Antoinette Swensson > > Univeristy of Washington/Harborview Medical Center > > Neuropathology > > _________________________________________________________________ > > > > [1]Find the music you love on MSN Music. Start downloading now! > > > > References > > > > 1. http://g.msn.com/8HMBENUS/2731??PS=47575 > > > > > > ------------------------------ > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > End of Histonet Digest, Vol 12, Issue 4 > > *************************************** > > > > > > > > ------------------------------ > > Message: 2 > Date: Wed, 3 Nov 2004 13:04:05 -0700 > From: "Elizabeth Chlipala" > Subject: [Histonet] TB block > To: > Message-ID: <000501c4c1e0$45969cb0$76d48a80@AMY> > Content-Type: text/plain; charset="us-ascii" > > Hello everyone > > I was wondering if there is anyone out there that could spare a block > that contains mycobacterium tuberculosis, and be willing to send that to > me. I would be willing to pay for the cost of shipping. I'm currently > trying to characterize an antibody to TB and the control block of > mycobacterium avium that I have will not work for my application. > > Thanks > > Liz > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > Premier Laboratory, LLC > P.O. Box 18592 > Boulder, Colorado 80308 > Office: (303) 735-5001 > Fax: (303) 735-3540 > liz@premierlab.com > www.premierlab.com > > Ship to Address: > Premier Laboratory > University of Colorado > MCDB, Room A3B40 > Boulder, Colorado 80309 > > > > ------------------------------ > > Message: 3 > Date: Wed, 3 Nov 2004 19:45:19 -0300 > From: "Hernan Aldana" > Subject: [Histonet] Fixation for carbohydrates > To: > Message-ID: <000001c4c1f6$cc542a40$389559c8@b3w6zzmtb6juvbs> > Content-Type: text/plain; charset="iso-8859-1" > > Hello all > I need the best fixation method for preserve hyaluronic acid in paraffin > sections. Formaldehyde, Bouin, Zencker, B5? Thanks in advance > > > Dr. Hern?n J. Aldana Marcos > Facultad de Medicina. Universidad de Mor?n > Machado 914. B1708JPD. Buenos Aires. Argentina > e-mail alternativo hernanjavier@yahoo.com > web: http://hjaldanamarcos.bravepages.com > http://www.ht.org.ar > > > > > > > ------------------------------ > > Message: 4 > Date: Wed, 3 Nov 2004 18:43:22 EST > From: JCarpenter764@aol.com > Subject: [Histonet] practical exam... > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset="US-ASCII" > > has anyone heard about the practical and wether they passed? > > > ------------------------------ > > Message: 5 > Date: Wed, 3 Nov 2004 17:49:34 -0600 > From: "maryjohnson" > Subject: [Histonet] exam > To: > Message-ID: <003501c4c1ff$c64cf510$b248bbd0@ugly> > Content-Type: text/plain; charset="iso-8859-1" > > Hi all I to am wating to hear about the practial exam. > > > ------------------------------ > > Message: 6 > Date: Wed, 03 Nov 2004 18:51:51 -0500 > From: Amos Brooks > Subject: [Histonet] Using expired reagents in IHC > To: histonet@lists.utsouthwestern.edu > Message-ID: <6.0.0.22.0.20041103182925.01afbe90@pop.earthlink.net> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Hi, > We always get new antibodies before the old expire. We only have > a > few expired ones kicking around. The Big High Muckety-Mucks (somewhat > understandably) have determined that the cost of the fines and dealing > with > battling the inspectors would be worse, in the short term, than the cost > of > purchasing just enough of the antibodies and not dealing with the extra > tech time involved in testing the same antibody monthly. > I can see the logic of either side of the coin. It may be easy to > > overlook a reagent quality gradually decreasing. Repeated testing of the > reagent costs time & money too. And the risk of the fines looming over > your > head is enough to make one hesitate. On the other hand the money saved on > keeping antibodies around for decades is appealing. I guess you could say > we're erring on the side of caution ... bloody expensive caution. > On another note, has anyone noticed that the listed shelf life of > > these reagents is shrinking lately. I just got one in today that has about > > 9 months until it expires. It is enough to make one wonder if the antibody > > companies are pushing for these regulations. I mean, really, where do they > > come up with these dates anyway? > Have a great day, > Amos Brooks > > > > At 10:54 AM 11/2/2004, you wrote: > >Message: 3 > >Date: Mon, 01 Nov 2004 14:26:32 -0500 > >From: "Richard Cartun" > >Subject: [Histonet] Using expired reagents in IHC > >To: > >Message-ID: > >Content-Type: text/plain; charset=US-ASCII > > > >It has been brought to my attention that the CAP Laboratory checklist > >has been revised as of 9/30/04 and now includes a question, "Are all > >immunohistochemical reagents used within their indicated expiration > >dates?". I have always believed that it is acceptable to use expired > >regents (mainly primary antibodies) as long as their reactivity is > >acceptable and documented. Obviously, the expiration date that > >manufactures use is not an accurate reflection of the reagent's > >likelihood of poor performance. Does anyone have information on this > >change? 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 > > > > > > ------------------------------ > > Message: 7 > Date: Wed, 3 Nov 2004 16:49:22 -0800 (PST) > From: Gareth Davis > Subject: [Histonet] Re: Exam > To: Histonet > Message-ID: <20041104004922.8210.qmail@web52703.mail.yahoo.com> > Content-Type: text/plain; charset=us-ascii > > >From what I've heard, the results for the pratical exam will not be out > until late November or early December. > Gareth > > > --------------------------------- > Do you Yahoo!? > Check out the new Yahoo! Front Page. www.yahoo.com/a > > ------------------------------ > > Message: 8 > Date: Wed, 3 Nov 2004 20:30:29 -0500 > From: "Denise Bland-Piontek" > Subject: [Histonet] test > To: > Message-ID: > <145F33B84793CF488578F86355B32E6912FBA4@mail.clinomicsbio.com> > Content-Type: text/plain; charset="iso-8859-1" > > > > > ------------------------------ > > Message: 9 > Date: Wed, 3 Nov 2004 20:29:39 EST > From: Dndsomi@aol.com > Subject: Re: [Histonet] Re: Exam > To: mrsgbd2001@yahoo.com, Histonet@lists.utsouthwestern.edu > Message-ID: <1a7.2a74b0d2.2ebae003@aol.com> > Content-Type: text/plain; charset="US-ASCII" > > I sent my practical in Sept. 2003 and got my results Jan. 15, 2004. > > > ------------------------------ > > Message: 10 > Date: Wed, 3 Nov 2004 21:54:23 EST > From: CrochiereSteve@aol.com > Subject: [Histonet] EGFR > To: PatPatterson@mhd.com, histonet@pathology.swmed.edu > Message-ID: <1d8.2f65199e.2ebaf3df@aol.com> > Content-Type: text/plain; charset="US-ASCII" > > Pat, > USLabs in California > > Steven M. Crochiere, HT(ASCP) > Histology Supervisor > LifePath Partners @ Mercy Medical Center > Springfield, MA 01104 > > > ------------------------------ > > Message: 11 > Date: Thu, 04 Nov 2004 13:48:12 +0000 > From: Chris.Goodall@bristol.ac.uk > Subject: [Histonet] Timm silver sulphide method for light microscopic > localisation of heavy metals. > To: histonet@lists.utsouthwestern.edu > Message-ID: <1099576092.418a331c8b602@webmail.bris.ac.uk> > Content-Type: text/plain > > > Dear All, > I have been asked to try the Timm method on some FFPE samples of > human brain. The papers I have read give methods for perfusing rat > brain with sodium sulphide solution followed by 3% gluteraldehyde in > 0.15M phosphate buffer, and further treatment with sodium sulphide > solution, or, post mortem brain samples snap frozen and frozen > sectioned. However health and safety rules here do not permit frozen > sectioning of unfixed human brain, or the use of gluteradehyde in the > embalming room, so I am going to attempt this method on formalin fixed > samples.My question is, has anyone experience of this method or any > suggestions? It is mentioned that autopsy material left in situ for one > or two days post mortem with no fixation may have enough endogenous > sulphide ions or sulphide groups in the tissue due to autolytic > activity and sulphide treatment may not be necessary, or if it is > necessary, does anyone have experience of timing in the sulphide > solution and is post fixation necessary after the initial formalin > fixation. The paper also mentions that to prevent loss of > metallosulphides present in the tissue the temperature of the wax > should not exceed 50C which means the use of the dreaded low > temperature wax, has anyone used conventional paraffin wax and got away > with it? > Sorry for the long request,and thank you, > Chris Goodall > > > > > ------------------------------ > > Message: 12 > Date: Thu, 4 Nov 2004 10:03:38 -0600 > From: ajennings@unmc.edu > Subject: [Histonet] Histonet 2005 Buttons > Cc: histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset=US-ASCII > > > > > > This is it...... I am going to let people continue to submit their designs > until next Friday and then I will be presenting the designs to our working > group to help me decide which one we will use for the Official 2005 > Histonet button. If you have been saving your stuff for last minute.....we > are in the 60 second count down for submissions. > > Please include a space for writing a name, the URL and a place for the > company who sponsors the buttons. > > Thanks a bunch for all the ones we have received it is fun to see the > talent and the involvement of the community > Anita > > > > > > ------------------------------ > > Message: 13 > Date: Thu, 4 Nov 2004 16:16:08 GMT > From: "mprice26@juno.com" > Subject: [Histonet] RE: Job Openings- Texarkana, TX > To: histonet@lists.utsouthwestern.edu > Message-ID: <20041104.081617.26898.15797@webmail28.nyc.untd.com> > Content-Type: text/plain > > > Looking for HT (ASCP) Registered Histotech at Pathology Services of > Texarkana. Great Location to raise a family. Day shift. > > Also looking for a temporary histotech for a couple of months. > > Contact Marsha Price @ mprice26@juno.com. > > Marsha > > ________________________________________________________________ > Juno Platinum $9.95. Juno SpeedBand $14.95. > Sign up for Juno Today at http://www.juno.com! > Look for special offers at Best Buy stores. > > > > ------------------------------ > > Message: 14 > Date: Thu, 04 Nov 2004 11:21:19 -0500 > From: John Kiernan > Subject: Re: [Histonet] Timm silver sulphide method for light > microscopiclocalisation of heavy metals. > To: Chris.Goodall@bristol.ac.uk > Cc: histonet@lists.utsouthwestern.edu > Message-ID: <418A56FF.69DDC8C6@uwo.ca> > Content-Type: text/plain; charset=us-ascii > > The perfusion of sodium sulphide is necessary > to precipitate metal ions (principally zinc), > which are otherwise soluble. Relying on autolysis > to generate sulphide and precipitate the zinc > doesn't make sense because the zinc ions would > have diffused away from their original synaptic > sites, and the tissue would be decomposing > anyway. Timm's method simply won't work (or not > in any meaningful way) on formaldehyde-fixed > paraffin-embedded specimens. > > Glutaraldehyde is not the only permissible > fixative after perfusing the buffered Na2S. > An alternative is to use an alcoholic fixative > that has been saturated with H2S gas. It's > unlikely that your health & safety regulations > will allow this if they won't let you take a > little jar of glutaraldehyde into the PM room! > > The best account of modern Timm staining is > probably Danscher,G & Zimmer,J (1978) > Histochemistry 55: 27-40. Danscher has also > published many more recent papers on > histochemical detection of zinc, mercury and > other metals with sulphide-silver methods. > > John Kiernan > London, Canada. > ___________________________ > Chris.Goodall@bristol.ac.uk wrote: > > > > Dear All, > > I have been asked to try the Timm method on some FFPE samples of > > human brain. The papers I have read give methods for perfusing rat > > brain with sodium sulphide solution followed by 3% gluteraldehyde in > > 0.15M phosphate buffer, and further treatment with sodium sulphide > > solution, or, post mortem brain samples snap frozen and frozen > > sectioned. However health and safety rules here do not permit frozen > > sectioning of unfixed human brain, or the use of gluteradehyde in the > > embalming room, so I am going to attempt this method on formalin fixed > > samples.My question is, has anyone experience of this method or any > > suggestions? It is mentioned that autopsy material left in situ for one > > or two days post mortem with no fixation may have enough endogenous > > sulphide ions or sulphide groups in the tissue due to autolytic > > activity and sulphide treatment may not be necessary, or if it is > > necessary, does anyone have experience of timing in the sulphide > > solution and is post fixation necessary after the initial formalin > > fixation. The paper also mentions that to prevent loss of > > metallosulphides present in the tissue the temperature of the wax > > should not exceed 50C which means the use of the dreaded low > > temperature wax, has anyone used conventional paraffin wax and got away > > with it? > > Sorry for the long request,and thank you, > > Chris Goodall > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 15 > Date: Thu, 4 Nov 2004 10:20:44 -0700 > From: "Patsy Ruegg" > Subject: RE: [Histonet] exam > To: "'maryjohnson'" , > > Message-ID: <000f01c4c292$a0e4f5f0$83020a0a@IHCTech> > Content-Type: text/plain; charset="us-ascii" > > We just did the grading of over 1100 slide sets for the HT exam last > weekend in Chicago, I am sure it will take some extra time this go round > to process all those. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > maryjohnson > Sent: Wednesday, November 03, 2004 4:50 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] exam > > > Hi all I to am wating to hear about the practial exam. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 16 > Date: Thu, 4 Nov 2004 10:32:45 -0700 > From: "Patsy Ruegg" > Subject: RE: [Histonet] BMP7 > To: "'Grant, Debra'" , > > Message-ID: <001101c4c294$4ba9e3f0$83020a0a@IHCTech> > Content-Type: text/plain; charset="us-ascii" > > Debra, > I have used Santa Cruz goat BMP-7 (L-19) sc-9305 on a lot of rat and > human samples but have not tried it on ms. SC lists it as working for > rat and h, since it is a goat poly it would be easy to try it on your > ms, ask SC if anyone has reported it as working on ms although I don't > usually find SC being very helpful for anything not listed being tried. > For rats I use it at 1:200 for 1 hr. using LSAB+ detection (I wish > someone would come up with a labeled polymer that works with goat > antibodies) and pepsin digestion (premade from Phoenix) at 37dc for two > changes 5 min. ea. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Grant, > Debra > Sent: Wednesday, November 03, 2004 10:47 AM > To: histonet@pathology.swmed.edu > Subject: [Histonet] BMP7 > > > > Has anyone used BMP7 antibody on mouse? If so, could you please send me > all the information, company, catalog #, protocol etc. > > Thanks in advance! > > Debby Grant > Research Technician II > Histology Core Facility > Stowers Institute for Medical Research > 1000 E. 50th Street > Kansas City, MO 64110 > drg@stowers-institute.org > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 17 > Date: Thu, 4 Nov 2004 10:48:19 -0700 > From: "Patsy Ruegg" > Subject: [Histonet] bone marrow mush > To: > Cc: ihcrg@neo.agsci.colostate.edu > Message-ID: <001701c4c296$79712e40$83020a0a@IHCTech> > Content-Type: text/plain; charset="us-ascii" > > Folks, > I have a question for you. One of my investigators harvested bone > marrow by just scrapping it out of the bone cavity, it is not in > anything (media, anticoagulant, fixative, etc.) they just stored it at > 4dc as a mush in a tube. They want me now to prepare it so that they > can see what cells they have. They also want to know if there are any > stem cells and what the viability is (by viability I mean they are > wondering if they could use it to grow some of the cells in culture). I > really don't think I can tell them if they can grow the cells or not. > Anyway I was thinking of trying to make a cell block from this material > and process into paraffin or maybe even do frozen sections on the > pellet. How would you go about making a pellet out of that stuff? > Would you put some buffer or media or fixative in the tube and spin it > down and then put it in HISTOGEL or wrap in tissue paper and then > process? > Patsy Ruegg > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 12, Issue 5 > *************************************** > > From lmormino <@t> surgipath.com Thu Nov 4 13:22:02 2004 From: lmormino <@t> surgipath.com (Linda Mormino) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] THANK YOU Message-ID: <000101c4c2a3$90aab1a0$30c9c08c@surgipath.com> To my friends and associates on the Histonet: My family and I sincerely appreciate all the cards and consolences we've received the past two weeks. This support has helped us through a very difficult period. We thank you from the bottom of our hearts. Ken Urban Surgipath Medical Ind. From joeamateur <@t> hotmail.com Thu Nov 4 13:45:01 2004 From: joeamateur <@t> hotmail.com (Jack England) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Viable radius (need a chemist!) Message-ID: Aloha Histonetters, I'm in a bind and I need some help. I'm trying to find/develop an assay that will determine the radius of viable tissue in a cross-section of FFPE tissue that is composed of living cells around a necrotic core. Standard nuclear stains (DAPI, hematoxylins, Sytox, etc) aren't enough, because they just show nuclei, and most stains I've looked at won't work because I'd have to fix and process the tissue before using them (thus killing all viable cells). I don't have access to a cryostat. My thinking so far is to incubate the tissue with viability reagents before fixation, then fix the tissue with the reagents in place. However, most of the mothods I've seen (acridine orange, calcein, etc.) involve reagents that are soluble in the solutions that are used in tissue processing. I thought about using a tetrazolium-salt metabolic assay (such as MTT), but since the formazan crystals that result are soluble in polar organic solvents (eg, ethanol and isopropanol), I'm pretty sure they'd wash out before I even got to sectioning. What I'd love to see is the insoluble crystal formation of a tetrazolium salt assay combined with the indiscriminate covalent bonding of a tyramide radical, so that my resulting chromagen (which indicates viable cells) won't wash out in ethanol, clearant, or water. Maybe a chomagen-conjugated tyramide would work (using endogenous peroxidases to my advantage, over a long incubation time)? Or maybe go with a MTT assay, but dehydrate in dry air with a dessicant, rather than a polar solvent (clearing in xylene as usual)? Can anyone help? I'm feeling like I'm right on the verge of cracking this, but doomed to fall just short because my organic chemistry isn't up to snuff. Thanks so much for your help! --With thanks and aloha, Jack England Tissue Genesis, Inc. http://www.tissuegenesis.com _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From making <@t> nersp.nerdc.ufl.edu Thu Nov 4 14:35:47 2004 From: making <@t> nersp.nerdc.ufl.edu (Mike King) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] [Fwd: Timm's staining human tissue] Message-ID: <418A92A3.6020001@nersp.nerdc.ufl.edu> -------- Original Message -------- Subject: Timm's staining human tissue Date: Thu, 04 Nov 2004 15:26:35 -0500 From: Mike King To: histonet-request@lists.utsouthwestern.edu Chris, As usual Dr. Kiernan is spot on in explaining why Timm's won't work in FFPE specimens. It is possible to do obtain acceptable Timm's histochemistry after immersion of fresh biopsy material in sulfide solutions, followed by immersion fixation. See our paper Murray KD, Isackson PJ, Eskin TA, King MA, Montesinos SP, Abraham LA, Roper SN. Altered mRNA expression for brain-derived neurotrophic factor and type II calcium/calmodulin-dependent protein kinase in the hippocampus of patients with intractable temporal lobe epilepsy. J Comp Neurol. 2000 Mar 20;418(4):411-22. I'll attach a pdf directly to you. The technique is likely to work foa autopsy tissue too. original post: From: Chris.Goodall@bristol.ac.uk Subject: [Histonet] Timm silver sulphide method for light microscopic localisation of heavy metals. To: histonet@lists.utsouthwestern.edu Message-ID: <1099576092.418a331c8b602@webmail.bris.ac.uk> Content-Type: text/plain Dear All, I have been asked to try the Timm method on some FFPE samples of human brain. The papers I have read give methods for perfusing rat brain with sodium sulphide solution followed by 3% gluteraldehyde in 0.15M phosphate buffer, and further treatment with sodium sulphide solution, or, post mortem brain samples snap frozen and frozen sectioned. However health and safety rules here do not permit frozen sectioning of unfixed human brain, or the use of gluteradehyde in the embalming room, so I am going to attempt this method on formalin fixed samples.My question is, has anyone experience of this method or any suggestions? It is mentioned that autopsy material left in situ for one or two days post mortem with no fixation may have enough endogenous sulphide ions or sulphide groups in the tissue due to autolytic activity and sulphide treatment may not be necessary, or if it is necessary, does anyone have experience of timing in the sulphide solution and is post fixation necessary after the initial formalin fixation. The paper also mentions that to prevent loss of metallosulphides present in the tissue the temperature of the wax should not exceed 50C which means the use of the dreaded low temperature wax, has anyone used conventional paraffin wax and got away with it? Sorry for the long request,and thank you, Chris Goodall -------- From joeamateur <@t> hotmail.com Thu Nov 4 14:36:35 2004 From: joeamateur <@t> hotmail.com (Jack England) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] a much easier chemistry question Message-ID: Aloha all, Here's a much more fun chemistry question for anyone that wants to bite: what is the best method to remove trypan/evans blue from cotton fabric? One of my colleagues got some on her pants the other day and asked what the best way to get it out was. I did not know, so I put some on cotton gauze, and tried: --acid alcohol...faded the stain a little, but didn't get rid of it --DI water...no change --absolute ethanol...no change --xylene...no change --ethanol/non-ionic detergent/DI water rinse...no change --citrus oil clearant...no change Given that her fabric is tan (and thus not white, and thus likely not bleach-able), can anyone out there in histo-land suggest a way of de-staining trypan-blue-stained fabric? I've been wondering about this for a while now and figured I'd pass it around. --Many thanks and aloha to all, Jack England Tissue Genesis, Inc. http://www.tissuegenesis.com _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From histobabs <@t> hotmail.com Thu Nov 4 15:26:14 2004 From: histobabs <@t> hotmail.com (B T) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] HMB50 and D2-40 Message-ID: Hi all, Is anyone out there doing melanoma clone HMB50 on the Ventana Benchmark? If so, what protocol are you using? I am having no success working up this antibody and at this point i'm not sure if there might be a problem eith the antibody itself. Also, is anyone using D2-40 on the Benchmark......I am getting staining but would like to punch it up a notch without having the tissue fall off the slide. Help please! Barb From terribraud <@t> msn.com Thu Nov 4 15:51:34 2004 From: terribraud <@t> msn.com (TERRI BRAUD) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Dumbing down the practical??? Message-ID: I have a question for all those on the list who have taken the HT practical. Why do you think that the practical exam does not cover the material that it used to? Having taken mine in 1981, and then, later as a supervisor, having trained 8 OJT's to sucessful completion of their certification, I have observed that the practical is not near the test that it used to be. There is no CNS tissue, no decalcified tissue, no submission of both a special stain and a H&E from the same block. Just one slide, one block, one stain. Also how is the effect of fully automated special stainers vs prepared kits vs "homemade" solutions being taken into account? There isn't even an informational question pertaining to this so that the ASCP could begin to see the impact of the technology in our HT learning process? Will the technician that can successfully load an autostainer be as effective in troubleshooting a stain as one who has learned every step by hand? Will it even be necessary in the future of Histology? Just curious to hear some opinions - Terri From mcauliff <@t> umdnj.edu Thu Nov 4 19:22:27 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] a much easier chemistry question In-Reply-To: References: Message-ID: <418AD5D3.3050801@umdnj.edu> Soap and water? Geoff Jack England wrote: > Aloha all, > > Here's a much more fun chemistry question for anyone that wants to > bite: what is the best method to remove trypan/evans blue from cotton > fabric? One of my colleagues got some on her pants the other day and > asked what the best way to get it out was. I did not know, so I put > some on cotton gauze, and tried: > > --acid alcohol...faded the stain a little, but didn't get rid of it > --DI water...no change > --absolute ethanol...no change > --xylene...no change > --ethanol/non-ionic detergent/DI water rinse...no change > --citrus oil clearant...no change > > Given that her fabric is tan (and thus not white, and thus likely not > bleach-able), can anyone out there in histo-land suggest a way of > de-staining trypan-blue-stained fabric? I've been wondering about this > for a while now and figured I'd pass it around. > > --Many thanks and aloha to all, > Jack England > Tissue Genesis, Inc. > http://www.tissuegenesis.com > > _________________________________________________________________ > Express yourself instantly with MSN Messenger! Download today - it's > FREE! hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ > > > _______________________________________________ > 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 andrae <@t> u.washington.edu Thu Nov 4 16:21:10 2004 From: andrae <@t> u.washington.edu (A. Erickson) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] a much easier chemistry question In-Reply-To: <418AD5D3.3050801@umdnj.edu> References: <418AD5D3.3050801@umdnj.edu> Message-ID: scissors, or a patch of the HistoNet Logo? andra On Thu, 4 Nov 2004, Geoff McAuliffe wrote: > Soap and water? > > Geoff > > Jack England wrote: > >> Aloha all, >> >> Here's a much more fun chemistry question for anyone that wants to bite: >> what is the best method to remove trypan/evans blue from cotton fabric? >> One of my colleagues got some on her pants the other day and asked what the >> best way to get it out was. I did not know, so I put some on cotton gauze, >> and tried: >> >> --acid alcohol...faded the stain a little, but didn't get rid of it >> --DI water...no change >> --absolute ethanol...no change >> --xylene...no change >> --ethanol/non-ionic detergent/DI water rinse...no change >> --citrus oil clearant...no change >> >> Given that her fabric is tan (and thus not white, and thus likely not >> bleach-able), can anyone out there in histo-land suggest a way of >> de-staining trypan-blue-stained fabric? I've been wondering about this for >> a while now and figured I'd pass it around. >> >> --Many thanks and aloha to all, >> Jack England >> Tissue Genesis, Inc. >> http://www.tissuegenesis.com >> >> _________________________________________________________________ >> Express yourself instantly with MSN Messenger! Download today - it's FREE! >> hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > -- > -- > ********************************************** > Geoff McAuliffe, Ph.D. > Neuroscience and Cell Biology > Robert Wood Johnson Medical School > 675 Hoes Lane, Piscataway, NJ 08854 > voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu > ********************************************** > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From KMB1904 <@t> aol.com Thu Nov 4 16:39:49 2004 From: KMB1904 <@t> aol.com (KMB1904@aol.com) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] CEU's Message-ID: <158.42ea12fd.2ebc09b5@aol.com> Does anyone know where you can get CEU credits for histology? Are we obligated to have so many a year? And if so, how many?? I have never seen this information written anywhere. From bwhitaker <@t> brownpathology.com Thu Nov 4 16:42:23 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] a much easier chemistry question In-Reply-To: Message-ID: <000001c4c2bf$8d56baf0$3601a8c0@brownpathology.net> How about just dying the whole garment? Or doing a "tie-dye". Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of A. Erickson Sent: Thursday, November 04, 2004 4:21 PM To: Geoff McAuliffe Cc: Histonet@lists.utsouthwestern.edu; Jack England Subject: Re: [Histonet] a much easier chemistry question scissors, or a patch of the HistoNet Logo? andra On Thu, 4 Nov 2004, Geoff McAuliffe wrote: > Soap and water? > > Geoff > > Jack England wrote: > >> Aloha all, >> >> Here's a much more fun chemistry question for anyone that wants to >> bite: >> what is the best method to remove trypan/evans blue from cotton fabric? >> One of my colleagues got some on her pants the other day and asked what the >> best way to get it out was. I did not know, so I put some on cotton gauze, >> and tried: >> >> --acid alcohol...faded the stain a little, but didn't get rid of it >> --DI water...no change --absolute ethanol...no change >> --xylene...no change >> --ethanol/non-ionic detergent/DI water rinse...no change >> --citrus oil clearant...no change >> >> Given that her fabric is tan (and thus not white, and thus likely not >> bleach-able), can anyone out there in histo-land suggest a way of >> de-staining trypan-blue-stained fabric? I've been wondering about this for >> a while now and figured I'd pass it around. >> >> --Many thanks and aloha to all, >> Jack England >> Tissue Genesis, Inc. >> http://www.tissuegenesis.com >> >> _________________________________________________________________ >> Express yourself instantly with MSN Messenger! Download today - it's >> FREE! >> hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > -- > -- > ********************************************** > Geoff McAuliffe, Ph.D. > Neuroscience and Cell Biology > Robert Wood Johnson Medical School > 675 Hoes Lane, Piscataway, NJ 08854 > voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu > ********************************************** > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Thu Nov 4 16:57:01 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Dumbing down the practical??? Message-ID: <566FB0B522443D43AF02D2ADBE35A6F001007EE3@UTHEVS3.mail.uthouston.edu> Terri I also share your frustration but also understand why this is probably done in this manner. The ideal situation would be if applicants went to a specific location, all received the same tissues, and then were given a list of slides to produce. It tests all individuals under the same set of circumstances. This would provide the necessary stress that makes a technician able to deal with real life situations. Currently the system compared IHC slides that are done by hand and those done by machine. This places those individuals who work in very small labs that cannot afford an automatic stainer at a disadvantage. The acquiring of tissues is also difficult, working in a lab mostly concerned with Mohs technique probably don't get to see many samples of uterus. It also eliminates the possibility of cheating by individuals sending in slides prepared by another individual. While I think that testing as I outlined above is the most desirable method it brings with it a lot of problems. To set up such a system is possible but in the United States would be a logistic nightmare. To even get the large amounts of material necessary for testing would be very difficult (could keep a herd of cows perhaps?). Considering the difficulties that would be involved I believe that the current system, while not perfect, is the best under the circumstances. Just my opinion. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of TERRI BRAUD Sent: Thursday, November 04, 2004 3:52 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dumbing down the practical??? I have a question for all those on the list who have taken the HT practical. Why do you think that the practical exam does not cover the material that it used to? Having taken mine in 1981, and then, later as a supervisor, having trained 8 OJT's to sucessful completion of their certification, I have observed that the practical is not near the test that it used to be. There is no CNS tissue, no decalcified tissue, no submission of both a special stain and a H&E from the same block. Just one slide, one block, one stain. Also how is the effect of fully automated special stainers vs prepared kits vs "homemade" solutions being taken into account? There isn't even an informational question pertaining to this so that the ASCP could begin to see the impact of the technology in our HT learning process? Will the technician that can successfully load an autostainer be as effective in troubleshooting a stain as one who has learned every step by hand? Will it even be necessary in the future of Histology? Just curious to hear some opinions - Terri _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From scoop <@t> mail.nih.gov Thu Nov 4 17:42:32 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] bone marrow mush In-Reply-To: <001701c4c296$79712e40$83020a0a@IHCTech> References: <001701c4c296$79712e40$83020a0a@IHCTech> Message-ID: Hi Patsy, I think the viability depends on how long the cells were in the tube at 4C. I harvest bone marrow by scraping or flushing it out of a bone and then I manipulate it, etc. for a few hours and the cells maintain morphology and viability. You can also do metabolic labeling on bone marrow (the cells are still viable) for hours after harvesting it this way. But if it's been 24 hours or more I would think the cells would be dead and autolyzing. You should probably tell them to make a smear or put them into fixative after harvest if they're going to be on ice for a long time. Another way to determine what the cell population in the sample is would be to do a western with cell specific markers for the populations of interest or FACS. But again, if the sample has been sitting at 4C for a long time I would think the cells would be dead and the proteins degraded, so no good for FACS or western. If the sample was frozen at -20C it would probably be fine for western, though. Sharon >Folks, >I have a question for you. One of my investigators harvested bone >marrow by just scrapping it out of the bone cavity, it is not in >anything (media, anticoagulant, fixative, etc.) they just stored it at >4dc as a mush in a tube. They want me now to prepare it so that they >can see what cells they have. They also want to know if there are any >stem cells and what the viability is (by viability I mean they are >wondering if they could use it to grow some of the cells in culture). I >really don't think I can tell them if they can grow the cells or not. >Anyway I was thinking of trying to make a cell block from this material >and process into paraffin or maybe even do frozen sections on the >pellet. How would you go about making a pellet out of that stuff? >Would you put some buffer or media or fixative in the tube and spin it >down and then put it in HISTOGEL or wrap in tissue paper and then >process? >Patsy Ruegg >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From jkiernan <@t> uwo.ca Thu Nov 4 17:59:39 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] a much easier chemistry question References: Message-ID: <418AC26B.98C6BB84@uwo.ca> Your unsuccessful efforts were all acidic, neutral or hydrophobic. Trypan and Evans blue are anionic dyes with large, very hydrophilic molecules. Alkaline solutions extract anionic dyes, and urea (high concentration) can disrupt the non-ionic, non-covalent bonding that holds large hydrophilic dye molecules to substrates such as cellulose (cotton). Try a strong urea solution (8M=48%) made alkaline with borax or washing soda. Geoff McAuliffe's simpler suggestion of soap and water may work, because soap is alkaline. Keep us informed. All who use dyes need this sort of information. (I have a grey tie with alcian blue spots dating from the 1960s; wouldn't want to remove them now, though I'm sure it would be impossible without dissolving the fabric.) John Kiernan London, Canada ------------------------------------- Jack England wrote: > > Aloha all, > > Here's a much more fun chemistry question for anyone that wants to bite: > what is the best method to remove trypan/evans blue from cotton fabric? One > of my colleagues got some on her pants the other day and asked what the best > way to get it out was. I did not know, so I put some on cotton gauze, and > tried: > > --acid alcohol...faded the stain a little, but didn't get rid of it > --DI water...no change > --absolute ethanol...no change > --xylene...no change > --ethanol/non-ionic detergent/DI water rinse...no change > --citrus oil clearant...no change > > Given that her fabric is tan (and thus not white, and thus likely not > bleach-able), can anyone out there in histo-land suggest a way of > de-staining trypan-blue-stained fabric? I've been wondering about this for a > while now and figured I'd pass it around. > > --Many thanks and aloha to all, > Jack England > Tissue Genesis, Inc. > http://www.tissuegenesis.com > > _________________________________________________________________ > Express yourself instantly with MSN Messenger! Download today - it's FREE! > hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mprice26 <@t> juno.com Thu Nov 4 19:41:35 2004 From: mprice26 <@t> juno.com (marsha r price) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] RE: 2 Positions - Northeast Texas Message-ID: <20041104.194135.5908.1.mprice26@juno.com> Dear histonetters, We are looking for 2 Histology Technicians on the day shift at Pathology Services of Texarkana. We are also interested in a temp person untill we fill these positions immediately. Please forward your resumes to: mprice26@juno.com Thank you. Marsha Price ________________________________________________________________ Juno Platinum $9.95. Juno SpeedBand $14.95. Sign up for Juno Today at http://www.juno.com! Look for special offers at Best Buy stores. From lpwenk <@t> sbcglobal.net Thu Nov 4 20:15:13 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] CEU's References: <158.42ea12fd.2ebc09b5@aol.com> Message-ID: <003001c4c2dd$4a635000$0ae1d445@domainnotset.invalid> Multiple faceted question. I'll try to answer it as best as I can. 1.There is NO national requirement to have CEU to continue to work in histology (or cytology or med tech, etc.). Most states have NO requirements (Florida and maybe California are exceptions). 2. Hospitals or your place of employment or even you lab itself can set any requirement they want, including no requirement. 3. ASCP has a new requirement for all people passing a certification exam after Jan. 1, 2004. Previously, it was - once certified = certified for life. Now there is a Certification Maintenance Program. (CMP) http://www.ascp.org/bor/cmp/index Now, if you want to maintain your certification status through ASCP, you must have 36 points of continuing education every 3 year, submit a form with this information along with a renewal fee (currently $50). Documentation such as CEU forms are not required, unless you are one of the randomly picked to be audited. Of the 36 points, for HT/HTL, 1 point must be on safety, 2 points in your area of specialty (i.e., histotechnology) and the rest of the points can be in your area of specialty, management, education, or any related lab area of interest. Points can be awarded for many types of CE, and each type can be worth different number of points. For example: - 1 hour workshop = 1 point - 1 hour teleconference = 1 point - 1 semester hour of college = 15 points - competency assessment by your employer = 2 points per year - authoring an article in a peer reviewed journal (such as NSH Journal of Histotechnology or ASCP Laboratory Medicine) = 5 points. 4. Ways to get CE: Check in the ASCP CMP packet (see above link, click on right side). Page 6 has lots of suggestions. - attending state meetings - attending national meetings - in-services held at your institution - teleconferences (available through NSH (301-262-6221) and ASCP (312-738-1336) - college classes (related to labs) - safety classes at your institution - competency assessment - present at state and national meetings - write articles for histotechnology journals - attend vendor workshops 36 points in 3 years really isn't a lot to have to get. And you don't have to spend a lot of money. In-services are free - just make certain you have documentation (i.e. sign in sheet). Most places have you do safety quizzes - fire, electrical, blood borne pathogens, etc. They would count. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Thursday, November 04, 2004 5:39 PM Subject: [Histonet] CEU's > Does anyone know where you can get CEU credits for histology? Are we > obligated to have so many a year? And if so, how many?? I have never seen this > information written anywhere. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Thu Nov 4 20:39:56 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Dumbing down the practical??? References: Message-ID: <003d01c4c2e0$bd883660$0ae1d445@domainnotset.invalid> I took my HTL exam in 1980, and have helped a lot of HT and HTL students since then take (and most) pass their exams. 1. In all those years, there has never been a time that both the H&E and a special stain has been requested on the same tissue. That would be double penalizing someone if they had a bad tissue. 2. There are 25 possible tissues that are listed on the complete HT practical list, with 48 stains on those tissues (some H&E, some specials). http://www.ascp.org/bor/getcertified/index.asp Bone is listed, as is spinal cord. (The HTL have bone, cerebellum and medulla. They also have 28 possible tissues with 61 possible stains.) The Histology Exam Committee randomly picks tissues from the lists, trying for varying degrees of difficulty of sectioning and staining. On this year's HT list of 9 tissues/stains, you are correct, there are no bone or CNS for the HT. There is uterus (which can be difficult to section due to density) and there is tonsil to be cut at 2-3 um, which is also difficult. At least these are the ones that my students had to struggle with this year. Also artery, until they got one without calcified plaque. (This year's HTL has bone and cervix which are dense, and kidney which had to be cut at 3 um.) As for staining, Retic can be tricky, as can carbol fuchsin. My students had to repeat these several times. PASH and Alcian blue were easier. H&E often revealed 2 shades of eosin, not 3 because they didn't pH it correctly. (My HTL students had problems with the PTAH stain on the muscle, the Giemsa on the bone, and the PAMS on the kidney. VVG on artery, Mucicarmine on small intestine and Pan-cytokeratin on cervix seemed to be easier for them.) As for automation, I'm sure this is playing an impact on the staining. Hopefully, someone on the Histology Exam Committee can address this one. Peggy A. Wenk, HTL(ASCP)SLS Program Director Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "TERRI BRAUD" To: Sent: Thursday, November 04, 2004 4:51 PM Subject: [Histonet] Dumbing down the practical??? > I have a question for all those on the list who have taken the HT practical. > Why do you think that the practical exam does not cover the material that > it used to? Having taken mine in 1981, and then, later as a supervisor, > having trained 8 OJT's to sucessful completion of their certification, I > have observed that the practical is not near the test that it used to be. > There is no CNS tissue, no decalcified tissue, no submission of both a > special stain and a H&E from the same block. Just one slide, one block, one > stain. Also how is the effect of fully automated special stainers vs > prepared kits vs "homemade" solutions being taken into account? There isn't > even an informational question pertaining to this so that the ASCP could > begin to see the impact of the technology in our HT learning process? Will > the technician that can successfully load an autostainer be as effective in > troubleshooting a stain as one who has learned every step by hand? Will it > even be necessary in the future of Histology? Just curious to hear some > opinions - Terri > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From wkreider <@t> u.washington.edu Thu Nov 4 20:44:43 2004 From: wkreider <@t> u.washington.edu (Wayne Kreider) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Re: histonet gas bubbles In-Reply-To: References: Message-ID: <418AE91B.7000403@u.washington.edu> Linda, Thanks for the input. I did get a chance to look at this paper briefly. Using these types of markers might be useful if we could see them with a high enough resolution. However, given my understanding of the method, we would have trouble directly differentiating a bubble from dissolved gases, as is our primary goal. I'm not just sure where we're headed with this in the future, but I'll let you know if we have some success. Wayne lkbauer@unmc.edu wrote: > > >Hello Wayne, >I recently read an article that may give you some direction. It explains >how some gases can be detected by use of trapping agents. This article on >nitric oxide( half-life of only a few seconds) summarizes recent methods >for visualizing NO in living tissues. Nitric Oxide 9 (2003) 217-228. Nitric >oxide imaging in living neuronal tissues using fluorscent probes. Author >Oliver von Bohen und Halbach. If you solve this I would be interested in >your method. Good luck! > > >Linda(Lin)Bauer >Department of Genetics, Cell Biology, and Anatomy >985455 Nebraska Medical Center >Omaha, NE 68198-5455 > >Phone: (402) 559-2863 >Fax: (402) 559-4001 >Email: lkbauer@unmc.edu > > > -- /**********************/ /Wayne Kreider/ /University of Washington/ /Applied Physics Lab / CIMU/ /106 Old Fisheries/ /206-543-1324/ From info <@t> ihcworld.com Thu Nov 4 22:22:58 2004 From: info <@t> ihcworld.com (IHC World) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] ImmPress In-Reply-To: Message-ID: Susan, I have been using ImmPRESS lately and did a parallol staining with HRP-Streptavidin (also from Vector). ImmPRESS is about 3-5 times more sensitive than HRP-Streptavidin which is reported more sensitive than Standard ABC method. A recent protocol developed is HIF-1a antibody staining. With LSAB method HIF-1a showed no staining, but with ImmPRESS showed strong nuclear staining pattern. Check image gallery as well for pictures of HIF-1a staining using ImmPRESS. Richard IHC World http://ihcworld.com -----Original Message----- From: Susan Travers [mailto:travers.3@osu.edu] Sent: Friday, October 29, 2004 4:54 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] ImmPress Has anyone used the new "ImmPress" reagent from Vector? If so, do you have a feel for how it compares to ABC in terms of sensitivity? Thanks Susan Travers -- Susan Travers, Ph.D. Professor, Oral Biology The Ohio State University College of Dentistry 305 W. 12th Avenue Columbus, Ohio 43210-1267 (614)292-6366(V) (614)247-6945(F) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bills <@t> icpmr.wsahs.nsw.gov.au Thu Nov 4 22:44:36 2004 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] HMB50 and D2-40 In-Reply-To: Message-ID: <000001c4c2f2$272e33b0$e1ce080a@wsahs.nsw.gov.au> bARB, I have spoken to the lady who does ourIHC on the Benchmark and she uses the HMB45 + 50 antibody from Neomarkers. She uses a mild CC1, the antibody diluted 1:50 and a 32min incubation time. Hope this helps. What is the other D2-40 antibody? Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of B T Sent: Friday, 05 November 2004 8:26 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HMB50 and D2-40 Hi all, Is anyone out there doing melanoma clone HMB50 on the Ventana Benchmark? If so, what protocol are you using? I am having no success working up this antibody and at this point i'm not sure if there might be a problem eith the antibody itself. Also, is anyone using D2-40 on the Benchmark......I am getting staining but would like to punch it up a notch without having the tissue fall off the slide. Help please! Barb _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From JMacDonald <@t> mtsac.edu Fri Nov 5 00:46:08 2004 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] CEU's Message-ID: California has no requirements for Histotechnicians for certifica tion or continuing education. Jennifer MacDonald -----histonet-bounces@lists.utsouthwestern. To: , From: Sent by: histonet-bounces@lists.u Date: 11/04/2004 06:15PM Subject: Re: [Histonet] CE Multiple faceted question. I'll can. 1.There is NO national requiremen work in histology (or cytology or med tech, requirements (Florida and maybe Califo 2. Hospitals or your place of employment or ev can set any requirement they want, including no requir 3. ASCP has a new requirement for all people passing a certif ication exam after Jan. 1, 2004. Previously, it was - once certified = certified for life. Now there is a Certification Maintenance Program. ( [1] Now, if you want to maintain y ASCP, you must have 36 points of contin a form with this information along w Documentation such as CEU forms are n randomly picked to be audited. Of be on safety, 2 points in your histotechnology) and the rest of the points can education, or any related lab Points can be awarded for many types of CE, and e worth different number of points. For example: - 1 - 1 hour teleconference = 1 point - 1 se - competency assessment by your emp - authoring an article in a peer reviewed jo Journal of Histotechnology or ASCP Laboratory Medicin 4. Ways to get CE: Check in the ASCP CMP pac side). Page 6 has lots of suggestion - attending state meetings - attending national meetings - - teleconferences (available throug (312-738-1336) - college classes (relat - safety classes at your institution - competency assessm - present at state and national meetings - write articles for his - attend vendor workshops 36 points in 3 ye don't have to spend a lot you have documentati safety quizzes - fire, Peggy A. W William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Thursday, November 04, 2004 5:39 PM Subjec > Does anyone know where you can ge Are we > obligated to have so man have never seen this &g > > > ______ ______________________ 5F__ > Histonet mailing l > Histonet@lists.utsouthwestern.edu > [2]http://li ___ ______________________ 5F__ Histonet maili Histonet@lists.utsouthwestern.edu [3]http://lists.ut < References 1. 3D"http://www.ascp.org/bor/cmp/index" 2. 3D"http://lis=/ 3. 3D"http://lists.uts=/ From Andrew.Prior <@t> Smith-Nephew.com Fri Nov 5 02:59:55 2004 From: Andrew.Prior <@t> Smith-Nephew.com (Prior, Andrew) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] RE: Alcian Blue 8GX - Thanks and replies Message-ID: As usual, an excellent response from all you Histonetters. Thank you very much for the info. It seems that there is still a big problem getting hold of Alcian Blue, so the price is still sky high! I have included the responses that were sent to me direct for general information. I have also talked to Surgipath (Europe) and they are trying to get their own supplier of the dye at the moment - could be useful in the future. Responses: 1) Try Rowley Chemical or Manufacturing Company. I buy a lot of my dyes from them. They specialize in histology dyes and stains. Frances L. Swain, HT(ASCP) A. A. S. Special Procedures Technician UAMS Center for Orthopaedic Research 4301 West Markham St., Slot 644 Little Rock AR 72205 www.rowleybio.com 2) Hi Andrew, The problem is the stuff is really hard to make safely and the few companies who did make it have stopped. Most of the stuff is made now by China and purchased from them. I know two companies here in the state that have it are Anatech and Polyscientific. Check their web sites. It will still be a lot more expensive than it was 3 years ago. Wish I could help more. Pam Marcum 3) Hello Andrew, A few years ago there was a reported worldwide shortage of AB 8GX. I think that this was because the major users (textile industry?) no longer required it and it is less economically viable to produce in the small quantities (relatively) used by histologists. Some companies such as Cellpath obtained supplies but it was expensive and limited to a certain amount per customer (?175 for 25g 3 years ago) I have not looked for a while but the Sigma price may well be an average price for it. I think there has been some discussion on this in the past on histonet so it may be worth looking at the archives. Dave Mehew Chief Biomedical Scientist Histopathology Department Queen's Hospital Burton Hospitals NHS Trust Burton upon Trent Staffordshire DE13 0RB England 4) Try the American company Anatech. http://www.anatechltdusa.com Their price is $96 for 25g. Still expensive, but it would make 2.5 litres of solution. Anatech's alcian blue is certified by the BSC. -- ------------------------------- 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 5) Dear Andrew, Anatech Ltd. sells Alcian Blue, 25 gram bottle for $95.55 US (estimate $52.01 UK pounds). It is certified by the Biological Stain Commission and has a dye content of 67%. Imperial Chemical Industries (ICI) of the UK was the manufacturer of Alcian Blue 8GX. ICI had trouble with the dye solubility for textile applications and various process changes in manufacturing were made in the 1950's and 1960's. Each different batch was given a different code. The codes 8GS and 8GX were found reliable for histology use. ICI ceased manufacturing in the 1970's and we all have been using for the last decades the leftover stock of ICI's original material. In 2001 ANATECH introduced their US manufactured Alcian Blue. Ordering information for our product is: Cat# 862 Product description: Alcian blue, 25 gram bottle Price: $95.55 US International orders require: Mastercard or VISA account # (to charge the cost of the product) Mastercard or VISA expiration date Name on the Mastercard or VISA card FedEx account (to charge shipping and duty costs) Shipping address Telephone number of recipient (as required by FedEx) Please do not hesitate to contact me if you have any additional questions. You may email me your ordering information and I will direct it though our ordering department. Sincerely, Ada 6) Andrew, Saw your enquiry on Histonet. We at CellPath can supply RHD-1040-25G - ALCIAN BLUE (CERTIFIED) - 25g - 95.00 GBP Plus ?5 carriage Hope this helps. Brian Reid Northern Teritory Manager CellPath PLC Unit 66 Mochdre Enterprise Park Newtown, Powys Mid Wales SY16 4LE Office Tel 01686 611 333 -------------------------------------------------- Hope you find these useful Andrew Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. From jim <@t> hteqa.co.uk Fri Nov 5 03:27:54 2004 From: jim <@t> hteqa.co.uk (Jim Elsam) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] a much easier chemistry question Message-ID: Jack Conn's Biological Stains, while giving no direct instruction on the removal of Evans blue, does conclude with the assertion that "light fastness is poor" so perhaps your colleague should sit at the window; or maybe a holiday in sunny climes is called for! Jim ~~~~~~~~~~~~~~~~~~~~~~ Jim Elsam, HTEQA Services Ltd From JWEEMS <@t> sjha.org Fri Nov 5 06:55:05 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] CEU's Message-ID: <83AACDB0810528418AA106F9AE9B7F7E50749E@sjhaexc02.sjha.org> 3. ASCP has a new requirement for all people passing a certification exam after Jan. 1, 2004. Previously, it was - once certified = certified for life. Now there is a Certification Maintenance Program. (CMP) Hi Peggy, This was news to me. Does this mean that all of us need to do this or just the ones certified after Jan. 1? Thanks, 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 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 n.cragg <@t> epistem.co.uk Fri Nov 5 06:58:57 2004 From: n.cragg <@t> epistem.co.uk (ncragg) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] HPV16 positive control Message-ID: <01C4C337.36D394F0.n.cragg@epistem.co.uk> Does anybody know where we can obtain HPV16 positive control slides for FFPE IHC? I am helping a medic with his research, who wants to stain patient samples with HPV16 - I have found a couple of HPV16 specific antibodies, but have suggested that we need a positive control. Dako sell HPV positive control slides although these are not specifically for HPV16. Would this be OK as a positive control or do we really need HPV16 specific? Thanks in advance, Nicola Cragg Nicola Cragg BSc Epistem Ltd Manchester, UK From GDawson <@t> Milw.Dynacare.com Fri Nov 5 07:57:18 2004 From: GDawson <@t> Milw.Dynacare.com (Dawson, Glen) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Getting out of Digest Format Message-ID: All, I can't handle any more people replying to the digest and having that complete digest attached to their reply!!!!! Sifting through this has become a real chore and it just keeps happening. Is there an easy way to get out of that digest format? I just visited the histonet "homepage" and couldn't find where I could do this. Suggestions would be appreciated. Thanx, Glen Dawson From pruegg <@t> ihctech.net Fri Nov 5 08:27:14 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Dumbing down the practical??? In-Reply-To: <003d01c4c2e0$bd883660$0ae1d445@domainnotset.invalid> Message-ID: These are all good issues raised concerning the HT exams. Seeing a lot of auto stained tissues submitted is probably one reason why starting in 2005 the examinee must pass the written before they submit the practical. Many of us are thinking that some of those who pass the practical may not know the theory and will not be able to pass the written and that requiring the written be passed first will weed out many. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of lpwenk@sbcglobal.net Sent: Thursday, November 04, 2004 7:40 PM To: TERRI BRAUD; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Dumbing down the practical??? I took my HTL exam in 1980, and have helped a lot of HT and HTL students since then take (and most) pass their exams. 1. In all those years, there has never been a time that both the H&E and a special stain has been requested on the same tissue. That would be double penalizing someone if they had a bad tissue. 2. There are 25 possible tissues that are listed on the complete HT practical list, with 48 stains on those tissues (some H&E, some specials). http://www.ascp.org/bor/getcertified/index.asp Bone is listed, as is spinal cord. (The HTL have bone, cerebellum and medulla. They also have 28 possible tissues with 61 possible stains.) The Histology Exam Committee randomly picks tissues from the lists, trying for varying degrees of difficulty of sectioning and staining. On this year's HT list of 9 tissues/stains, you are correct, there are no bone or CNS for the HT. There is uterus (which can be difficult to section due to density) and there is tonsil to be cut at 2-3 um, which is also difficult. At least these are the ones that my students had to struggle with this year. Also artery, until they got one without calcified plaque. (This year's HTL has bone and cervix which are dense, and kidney which had to be cut at 3 um.) As for staining, Retic can be tricky, as can carbol fuchsin. My students had to repeat these several times. PASH and Alcian blue were easier. H&E often revealed 2 shades of eosin, not 3 because they didn't pH it correctly. (My HTL students had problems with the PTAH stain on the muscle, the Giemsa on the bone, and the PAMS on the kidney. VVG on artery, Mucicarmine on small intestine and Pan-cytokeratin on cervix seemed to be easier for them.) As for automation, I'm sure this is playing an impact on the staining. Hopefully, someone on the Histology Exam Committee can address this one. Peggy A. Wenk, HTL(ASCP)SLS Program Director Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "TERRI BRAUD" To: Sent: Thursday, November 04, 2004 4:51 PM Subject: [Histonet] Dumbing down the practical??? > I have a question for all those on the list who have taken the HT practical. > Why do you think that the practical exam does not cover the material that > it used to? Having taken mine in 1981, and then, later as a supervisor, > having trained 8 OJT's to sucessful completion of their certification, I > have observed that the practical is not near the test that it used to be. > There is no CNS tissue, no decalcified tissue, no submission of both a > special stain and a H&E from the same block. Just one slide, one block, one > stain. Also how is the effect of fully automated special stainers vs > prepared kits vs "homemade" solutions being taken into account? There isn't > even an informational question pertaining to this so that the ASCP could > begin to see the impact of the technology in our HT learning process? Will > the technician that can successfully load an autostainer be as effective in > troubleshooting a stain as one who has learned every step by hand? Will it > even be necessary in the future of Histology? Just curious to hear some > opinions - Terri > > > > _______________________________________________ > Histonet mailing list > Histonet@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 ian.montgomery <@t> bio.gla.ac.uk Fri Nov 5 08:54:56 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Zinc. Message-ID: <6.1.2.0.2.20041105145141.02ea08b0@udcf.gla.ac.uk> When choosing an antibody the companies usually give the preferred method of preparation, frozen, formalin, zinc etc. When they say zinc, do they mean a zinc only fixative or a zinc formalin mixture? Ian. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk From LINDA.MARGRAF <@t> childrens.com Fri Nov 5 08:39:57 2004 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Getting out of Digest Format Message-ID: Dear Glen To switch out of digest mode you have to go to http://lists.utsouthwestern.edu/mailman/listinfo/histonet You have to scroll almost to the botttom of the page, under the Histonet subscriber header, where you put in your email address. You will then go to a field that asks for you password (you can request it if you can't remember) then you will go to a field where you can switch the digest function on or off (you will want to go to off) . There are also options to block duplicate messages, unsubscribe or change your address or password. Please let me know if you have problems. Linda M Histonet administrator From Rcartun <@t> harthosp.org Fri Nov 5 09:01:52 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] p34 IHC Message-ID: Is anyone doing IHC for "p34" on formalin-fixed tissue for dysplasia? If so, where do you obtain your antibody? Thank you and have a great day! 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 Jackie.O'Connor <@t> abbott.com Fri Nov 5 09:10:28 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] Zinc. Message-ID: I've been forced to use a zinc only fixative (Streck Tissue Fixative) for a couple of years - BD (Pharmingen) recommends the use of this for many of it's antibodies - i.e. Caspase-3 and CD31 - works great, but the morphology stinks. I'm in the process of 'crossing over' to find antibodies that will work on FFPE as well on my xenografts. Jackie O' Ian Montgomery Sent by: histonet-bounces@lists.utsouthwestern.edu 11/05/2004 08:54 AM To: histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Zinc. When choosing an antibody the companies usually give the preferred method of preparation, frozen, formalin, zinc etc. When they say zinc, do they mean a zinc only fixative or a zinc formalin mixture? Ian. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brett_connolly <@t> merck.com Fri Nov 5 09:36:38 2004 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] GLUT1 antibody Message-ID: All, Looking for antibodies to GLUT 1 for human tissue. Any recommendations? Thanks, Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (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 Janet.Bonner <@t> FLHOSP.ORG Fri Nov 5 09:54:08 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] CEU's Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB40D8@fh2k093.fhmis.net> Well, Florida sure does - I order mine through Anderson Continuing Education ; 3268 Ramos Circle / P.O.Box 276297 ; Sacramento, Ca 95827 Phone: 1-800-532-2332 / FAX 1-916-856-5066 They're really easy to use. You buy the book and they send a test booklet of multiple choice questions and a scantron sheet - which you can fax when you're done. You'll get a certificate in the mail within a week. Each course is mostly 24 CEUs with some exceptions that are 12 or 36. There's also one out of Miami, FL but I don't have the info on that. Give them a call!! Janet -----Original Message----- From: Jennifer MacDonald To: lpwenk@sbcglobal.net Cc: KMB1904@aol.com; histonet@pathology.swmed.edu Sent: 11/5/2004 1:46 AM Subject: Re: [Histonet] CEU's California has no requirements for Histotechnicians for certifica tion or continuing education. Jennifer MacDonald -----histonet-bounces@lists.utsouthwestern.= edu wrote: ----- To: , From: Sent by: histonet-bounces@lists.u= tsouthwestern.edu Date: 11/04/2004 06:15PM Subject: Re: [Histonet] CE= U's Multiple faceted question. I'll = try to answer it as best as I can. 1.There is NO national requiremen= t to have CEU to continue to work in histology (or cytology or med tech,= etc.). Most states have NO requirements (Florida and maybe Califo= rnia are exceptions). 2. Hospitals or your place of employment or ev= en you lab itself can set any requirement they want, including no requir= ement. 3. ASCP has a new requirement for all people passing a certif ication exam after Jan. 1, 2004. Previously, it was - once certified = certified for life. Now there is a Certification Maintenance Program. (= CMP) [1]= http://www.ascp.org/bor/cmp/index Now, if you want to maintain y= our certification status through ASCP, you must have 36 points of contin= uing education every 3 year, submit a form with this information along w= ith a renewal fee (currently $50). Documentation such as CEU forms are n= ot required, unless you are one of the randomly picked to be audited. Of= the 36 points, for HT/HTL, 1 point must be on safety, 2 points in your = area of specialty (i.e., histotechnology) and the rest of the points can= be in your area of specialty, management, education, or any related lab= area of interest. Points can be awarded for many types of CE, and e= ach type can be worth different number of points. For example: - 1= hour workshop =3D 1 point - 1 hour teleconference = 1 point - 1 se= mester hour of college =3D 15 points - competency assessment by your emp= loyer =3D 2 points per year - authoring an article in a peer reviewed jo= urnal (such as NSH Journal of Histotechnology or ASCP Laboratory Medicin= e) =3D 5 points. 4. Ways to get CE: Check in the ASCP CMP pac= ket (see above link, click on right side). Page 6 has lots of suggestion= s. - attending state meetings - attending national meetings - = in-services held at your institution - teleconferences (available throug= h NSH (301-262-6221) and ASCP (312-738-1336) - college classes (relat= ed to labs) - safety classes at your institution - competency assessm= ent - present at state and national meetings - write articles for his= totechnology journals - attend vendor workshops 36 points in 3 ye= ars really isn't a lot to have to get. And you don't have to spend a lot= of money. In-services are free - just make certain you have documentati= on (i.e. sign in sheet). Most places have you do safety quizzes - fire, = electrical, blood borne pathogens, etc. They would count. Peggy A. W= enk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Thursday, November 04, 2004 5:39 PM Subjec= t: [Histonet] CEU's > Does anyone know where you can ge= t CEU credits for histology? Are we > obligated to have so man= y a year? And if so, how many?? I have never seen this &g= t; information written anywhere. > > > ______ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F > Histonet mailing l= ist > Histonet@lists.utsouthwestern.edu > [2]http://li= sts.utsouthwestern.edu/mailman/listinfo/histonet ___ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F Histonet maili= ng list Histonet@lists.utsouthwestern.edu [3]http://lists.ut= southwestern.edu/mailman/listinfo/histonet <= /FONT> References 1. 3D"http://www.ascp.org/bor/cmp/index" 2. 3D"http://lis=/ 3. 3D"http://lists.uts=/ _______________________________________________ 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 bwhitaker <@t> brownpathology.com Fri Nov 5 10:22:44 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] CEU's In-Reply-To: <07AB60D5D7B9754EBF56F360F98D083DEB40D8@fh2k093.fhmis.net> Message-ID: <000201c4c353$aec84980$3601a8c0@brownpathology.net> How many folks out there plan on doing the optional (for those already registered) Certification Maintenance Program? I think it's a good idea, and I plan on doing it, but I just wanted to know what the general feeling is. Will anyone require that of their employees? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: Friday, November 05, 2004 9:54 AM To: 'Jennifer MacDonald '; 'lpwenk@sbcglobal.net ' Cc: 'KMB1904@aol.com '; 'histonet@pathology.swmed.edu ' Subject: RE: [Histonet] CEU's Well, Florida sure does - I order mine through Anderson Continuing Education ; 3268 Ramos Circle / P.O.Box 276297 ; Sacramento, Ca 95827 Phone: 1-800-532-2332 / FAX 1-916-856-5066 They're really easy to use. You buy the book and they send a test booklet of multiple choice questions and a scantron sheet - which you can fax when you're done. You'll get a certificate in the mail within a week. Each course is mostly 24 CEUs with some exceptions that are 12 or 36. There's also one out of Miami, FL but I don't have the info on that. Give them a call!! Janet -----Original Message----- From: Jennifer MacDonald To: lpwenk@sbcglobal.net Cc: KMB1904@aol.com; histonet@pathology.swmed.edu Sent: 11/5/2004 1:46 AM Subject: Re: [Histonet] CEU's California has no requirements for Histotechnicians for certifica tion or continuing education. Jennifer MacDonald -----histonet-bounces@lists.utsouthwestern.= edu wrote: ----- To: , From: Sent by: histonet-bounces@lists.u= tsouthwestern.edu Date: 11/04/2004 06:15PM Subject: Re: [Histonet] CE= U's Multiple faceted question. I'll = try to answer it as best as I can. 1.There is NO national requiremen= t to have CEU to continue to work in histology (or cytology or med tech,= etc.). Most states have NO requirements (Florida and maybe Califo= rnia are exceptions). 2. Hospitals or your place of employment or ev= en you lab itself can set any requirement they want, including no requir= ement. 3. ASCP has a new requirement for all people passing a certif ication exam after Jan. 1, 2004. Previously, it was - once certified = certified for life. Now there is a Certification Maintenance Program. (= CMP) [1]= http://www.ascp.org/bor/cmp/index Now, if you want to maintain y= our certification status through ASCP, you must have 36 points of contin= uing education every 3 year, submit a form with this information along w= ith a renewal fee (currently $50). Documentation such as CEU forms are n= ot required, unless you are one of the randomly picked to be audited. Of= the 36 points, for HT/HTL, 1 point must be on safety, 2 points in your = area of specialty (i.e., histotechnology) and the rest of the points can= be in your area of specialty, management, education, or any related lab= area of interest. Points can be awarded for many types of CE, and e= ach type can be worth different number of points. For example: - 1= hour workshop =3D 1 point - 1 hour teleconference = 1 point - 1 se= mester hour of college =3D 15 points - competency assessment by your emp= loyer =3D 2 points per year - authoring an article in a peer reviewed jo= urnal (such as NSH Journal of Histotechnology or ASCP Laboratory Medicin= e) =3D 5 points. 4. Ways to get CE: Check in the ASCP CMP pac= ket (see above link, click on right side). Page 6 has lots of suggestion= s. - attending state meetings - attending national meetings - = in-services held at your institution - teleconferences (available throug= h NSH (301-262-6221) and ASCP (312-738-1336) - college classes (relat= ed to labs) - safety classes at your institution - competency assessm= ent - present at state and national meetings - write articles for his= totechnology journals - attend vendor workshops 36 points in 3 ye= ars really isn't a lot to have to get. And you don't have to spend a lot= of money. In-services are free - just make certain you have documentati= on (i.e. sign in sheet). Most places have you do safety quizzes - fire, = electrical, blood borne pathogens, etc. They would count. Peggy A. W= enk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Thursday, November 04, 2004 5:39 PM Subjec= t: [Histonet] CEU's > Does anyone know where you can ge= t CEU credits for histology? Are we > obligated to have so man= y a year? And if so, how many?? I have never seen this &g= t; information written anywhere. > > > ______ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F > Histonet mailing l= ist > Histonet@lists.utsouthwestern.edu > [2]http://li= sts.utsouthwestern.edu/mailman/listinfo/histonet ___ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F Histonet maili= ng list Histonet@lists.utsouthwestern.edu [3]http://lists.ut= southwestern.edu/mailman/listinfo/histonet <= /FONT> References 1. 3D"http://www.ascp.org/bor/cmp/index" 2. 3D"http://lis=/ 3. 3D"http://lists.uts=/ _______________________________________________ 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 MAUGER <@t> email.chop.edu Fri Nov 5 10:23:45 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] GLUT1 antibody Message-ID: Brett, I use a good Glut-1 from Dako. Its a rabbit anti human works well in formalin fixed tissue with heat retrieval. Cat # A3536. Jo >>> "Connolly, Brett M" 11/05/04 10:36AM >>> All, Looking for antibodies to GLUT 1 for human tissue. Any recommendations? Thanks, Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From POWELL_SA <@t> Mercer.edu Fri Nov 5 10:38:11 2004 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] CEU's In-Reply-To: <000201c4c353$aec84980$3601a8c0@brownpathology.net> Message-ID: I plan to participate. I too think it is a good idea. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bonnie Whitaker Sent: Friday, November 05, 2004 11:23 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] CEU's How many folks out there plan on doing the optional (for those already registered) Certification Maintenance Program? I think it's a good idea, and I plan on doing it, but I just wanted to know what the general feeling is. Will anyone require that of their employees? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: Friday, November 05, 2004 9:54 AM To: 'Jennifer MacDonald '; 'lpwenk@sbcglobal.net ' Cc: 'KMB1904@aol.com '; 'histonet@pathology.swmed.edu ' Subject: RE: [Histonet] CEU's Well, Florida sure does - I order mine through Anderson Continuing Education ; 3268 Ramos Circle / P.O.Box 276297 ; Sacramento, Ca 95827 Phone: 1-800-532-2332 / FAX 1-916-856-5066 They're really easy to use. You buy the book and they send a test booklet of multiple choice questions and a scantron sheet - which you can fax when you're done. You'll get a certificate in the mail within a week. Each course is mostly 24 CEUs with some exceptions that are 12 or 36. There's also one out of Miami, FL but I don't have the info on that. Give them a call!! Janet -----Original Message----- From: Jennifer MacDonald To: lpwenk@sbcglobal.net Cc: KMB1904@aol.com; histonet@pathology.swmed.edu Sent: 11/5/2004 1:46 AM Subject: Re: [Histonet] CEU's California has no requirements for Histotechnicians for certifica tion or continuing education. Jennifer MacDonald -----histonet-bounces@lists.utsouthwestern.= edu wrote: ----- To: , From: Sent by: histonet-bounces@lists.u= tsouthwestern.edu Date: 11/04/2004 06:15PM Subject: Re: [Histonet] CE= U's Multiple faceted question. I'll = try to answer it as best as I can. 1.There is NO national requiremen= t to have CEU to continue to work in histology (or cytology or med tech,= etc.). Most states have NO requirements (Florida and maybe Califo= rnia are exceptions). 2. Hospitals or your place of employment or ev= en you lab itself can set any requirement they want, including no requir= ement. 3. ASCP has a new requirement for all people passing a certif ication exam after Jan. 1, 2004. Previously, it was - once certified = certified for life. Now there is a Certification Maintenance Program. (= CMP) [1]= http://www.ascp.org/bor/cmp/index Now, if you want to maintain y= our certification status through ASCP, you must have 36 points of contin= uing education every 3 year, submit a form with this information along w= ith a renewal fee (currently $50). Documentation such as CEU forms are n= ot required, unless you are one of the randomly picked to be audited. Of= the 36 points, for HT/HTL, 1 point must be on safety, 2 points in your = area of specialty (i.e., histotechnology) and the rest of the points can= be in your area of specialty, management, education, or any related lab= area of interest. Points can be awarded for many types of CE, and e= ach type can be worth different number of points. For example: - 1= hour workshop =3D 1 point - 1 hour teleconference = 1 point - 1 se= mester hour of college =3D 15 points - competency assessment by your emp= loyer =3D 2 points per year - authoring an article in a peer reviewed jo= urnal (such as NSH Journal of Histotechnology or ASCP Laboratory Medicin= e) =3D 5 points. 4. Ways to get CE: Check in the ASCP CMP pac= ket (see above link, click on right side). Page 6 has lots of suggestion= s. - attending state meetings - attending national meetings - = in-services held at your institution - teleconferences (available throug= h NSH (301-262-6221) and ASCP (312-738-1336) - college classes (relat= ed to labs) - safety classes at your institution - competency assessm= ent - present at state and national meetings - write articles for his= totechnology journals - attend vendor workshops 36 points in 3 ye= ars really isn't a lot to have to get. And you don't have to spend a lot= of money. In-services are free - just make certain you have documentati= on (i.e. sign in sheet). Most places have you do safety quizzes - fire, = electrical, blood borne pathogens, etc. They would count. Peggy A. W= enk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Thursday, November 04, 2004 5:39 PM Subjec= t: [Histonet] CEU's > Does anyone know where you can ge= t CEU credits for histology? Are we > obligated to have so man= y a year? And if so, how many?? I have never seen this &g= t; information written anywhere. > > > ______ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F > Histonet mailing l= ist > Histonet@lists.utsouthwestern.edu > [2]http://li= sts.utsouthwestern.edu/mailman/listinfo/histonet ___ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F Histonet maili= ng list Histonet@lists.utsouthwestern.edu [3]http://lists.ut= southwestern.edu/mailman/listinfo/histonet <= /FONT> References 1. 3D"http://www.ascp.org/bor/cmp/index" 2. 3D"http://lis=/ 3. 3D"http://lists.uts=/ _______________________________________________ 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 From scoop <@t> mail.nih.gov Fri Nov 5 10:45:43 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] erythropoietin receptor Message-ID: Hi, I'm interested in doing IHC for erythropoietin receptor in FFPE tissues, especially in brain. I've tried to do this unsuccessfully in the past. Does anyone know if this can be done, are there any special things I need to do (antigen retrieval, etc.?) and can anyone recommend antibodies for this purpose? (I'm using mouse tissue) Thanks a lot! Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From rbarnhart <@t> summithealth.org Fri Nov 5 10:49:25 2004 From: rbarnhart <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] CEU's Message-ID: I think it is a good idea and also am planning on doing it. I would not be opposed if my employeer requires it for the entire lab staff (MT, MLT, CT and the 1 phelb that we have) >>> "Bonnie Whitaker" 11/5/2004 11:22:44 AM >>> How many folks out there plan on doing the optional (for those already registered) Certification Maintenance Program? I think it's a good idea, and I plan on doing it, but I just wanted to know what the general feeling is. Will anyone require that of their employees? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: Friday, November 05, 2004 9:54 AM To: 'Jennifer MacDonald '; 'lpwenk@sbcglobal.net ' Cc: 'KMB1904@aol.com '; 'histonet@pathology.swmed.edu ' Subject: RE: [Histonet] CEU's Well, Florida sure does - I order mine through Anderson Continuing Education ; 3268 Ramos Circle / P.O.Box 276297 ; Sacramento, Ca 95827 Phone: 1-800-532-2332 / FAX 1-916-856-5066 They're really easy to use. You buy the book and they send a test booklet of multiple choice questions and a scantron sheet - which you can fax when you're done. You'll get a certificate in the mail within a week. Each course is mostly 24 CEUs with some exceptions that are 12 or 36. There's also one out of Miami, FL but I don't have the info on that. Give them a call!! Janet -----Original Message----- From: Jennifer MacDonald To: lpwenk@sbcglobal.net Cc: KMB1904@aol.com; histonet@pathology.swmed.edu Sent: 11/5/2004 1:46 AM Subject: Re: [Histonet] CEU's California has no requirements for Histotechnicians for certifica tion or continuing education. Jennifer MacDonald -----histonet-bounces@lists.utsouthwestern.= edu wrote: ----- To: , From: Sent by: histonet-bounces@lists.u= tsouthwestern.edu Date: 11/04/2004 06:15PM Subject: Re: [Histonet] CE= U's Multiple faceted question. I'll = try to answer it as best as I can. 1.There is NO national requiremen= t to have CEU to continue to work in histology (or cytology or med tech,= etc.). Most states have NO requirements (Florida and maybe Califo= rnia are exceptions). 2. Hospitals or your place of employment or ev= en you lab itself can set any requirement they want, including no requir= ement. 3. ASCP has a new requirement for all people passing a certif ication exam after Jan. 1, 2004. Previously, it was - once certified = certified for life. Now there is a Certification Maintenance Program. (= CMP) [1]= http://www.ascp.org/bor/cmp/index Now, if you want to maintain y= our certification status through ASCP, you must have 36 points of contin= uing education every 3 year, submit a form with this information along w= ith a renewal fee (currently $50). Documentation such as CEU forms are n= ot required, unless you are one of the randomly picked to be audited. Of= the 36 points, for HT/HTL, 1 point must be on safety, 2 points in your = area of specialty (i.e., histotechnology) and the rest of the points can= be in your area of specialty, management, education, or any related lab= area of interest. Points can be awarded for many types of CE, and e= ach type can be worth different number of points. For example: - 1= hour workshop =3D 1 point - 1 hour teleconference = 1 point - 1 se= mester hour of college =3D 15 points - competency assessment by your emp= loyer =3D 2 points per year - authoring an article in a peer reviewed jo= urnal (such as NSH Journal of Histotechnology or ASCP Laboratory Medicin= e) =3D 5 points. 4. Ways to get CE: Check in the ASCP CMP pac= ket (see above link, click on right side). Page 6 has lots of suggestion= s. - attending state meetings - attending national meetings - = in-services held at your institution - teleconferences (available throug= h NSH (301-262-6221) and ASCP (312-738-1336) - college classes (relat= ed to labs) - safety classes at your institution - competency assessm= ent - present at state and national meetings - write articles for his= totechnology journals - attend vendor workshops 36 points in 3 ye= ars really isn't a lot to have to get. And you don't have to spend a lot= of money. In-services are free - just make certain you have documentati= on (i.e. sign in sheet). Most places have you do safety quizzes - fire, = electrical, blood borne pathogens, etc. They would count. Peggy A. W= enk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Thursday, November 04, 2004 5:39 PM Subjec= t: [Histonet] CEU's > Does anyone know where you can ge= t CEU credits for histology? Are we > obligated to have so man= y a year? And if so, how many?? I have never seen this &g= t; information written anywhere. > > > ______ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F > Histonet mailing l= ist > Histonet@lists.utsouthwestern.edu > [2]http://li= sts.utsouthwestern.edu/mailman/listinfo/histonet ___ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F Histonet maili= ng list Histonet@lists.utsouthwestern.edu [3]http://lists.ut= southwestern.edu/mailman/listinfo/histonet <= /FONT> References 1. 3D"http://www.ascp.org/bor/cmp/index" 2. 3D"http://lis=/ 3. 3D"http://lists.uts=/ _______________________________________________ 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 From histobabs <@t> hotmail.com Fri Nov 5 11:04:35 2004 From: histobabs <@t> hotmail.com (B T) Date: Fri Sep 16 15:24:15 2005 Subject: [Histonet] HMB50 and D2-40 Message-ID: thank you.......we have tried that protocol as well as a host of others....I have just retrieved offline and stained by hand as well....all to no avail!! I think the antibody may be a dud.....since we have tried just about everything apart from standing on one foot and singing Jingle Bells while the slides "stain". Thanks for the info....it is very helpful. The D2-40 is supposed to be a vascular marker....this is what the ordering pathologist intends to use it for. Thanks again for your help......this supports my feeling that we got a bad batch of antibody. Barb >From: "Bill Sinai" >To: "'B T'" ,"histonet (E-mail)" > >Subject: RE: [Histonet] HMB50 and D2-40 >Date: Fri, 5 Nov 2004 15:44:36 +1100 > > >bARB, > >I have spoken to the lady who does ourIHC on the Benchmark and she uses the >HMB45 + 50 antibody from Neomarkers. She uses a mild CC1, the antibody >diluted 1:50 and a 32min incubation time. > >Hope this helps. > >What is the other D2-40 antibody? > >Bill Sinai >Laboratory Manager >Tissue Pathology, ICPMR >Westmead NSW 2145 >Australia >Ph 02 9845 7774 > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of B T >Sent: Friday, 05 November 2004 8:26 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] HMB50 and D2-40 > > >Hi all, > >Is anyone out there doing melanoma clone HMB50 on the Ventana Benchmark? If >so, what protocol are you using? I am having no success working up this >antibody and at this point i'm not sure if there might be a problem eith >the >antibody itself. > >Also, is anyone using D2-40 on the Benchmark......I am getting staining but >would like to punch it up a notch without having the tissue fall off the >slide. > >Help please! > >Barb > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >__________________________________________________________________ > >This electronic message and any attachments may be confidential. If you >are not the intended recipient of this message would you please delete the >message and any attachments and advise the sender. Western Sydney >Area Health Services (WSAHS) uses virus scanning software but excludes >any liability for viruses contained in any email or attachment. > >This email may contain privileged and confidential information intended >only for the use of the addressees named above. If you are not the >intended recipient of this email, you are hereby notified that any use, >dissemination, distribution, or reproduction of this email is prohibited. >If >you have received this email in error, please notify WSAHS >immediately. > >Any views expressed in this email are those of the individual sender >except where the sender expressly and with authority states them >to be the views of WSAHS. From Janet.Bonner <@t> FLHOSP.ORG Fri Nov 5 11:07:01 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] CEU's Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB40DA@fh2k093.fhmis.net> It's a good idea, especially if you go state to state, or just need an update. I've been in several states and heard talk of starting the CEU program in all these locals. AND I would not be surprised if us "oldsters" won't be required to do it in the future! I've taken a couple of courses that were good updates - the Histology field is no longer stagnant as it seemed to be from 1940-1970. -----Original Message----- From: Rebecca Barnhart [mailto:rbarnhart@summithealth.org] Sent: Friday, November 05, 2004 11:49 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] CEU's I think it is a good idea and also am planning on doing it. I would not be opposed if my employeer requires it for the entire lab staff (MT, MLT, CT and the 1 phelb that we have) >>> "Bonnie Whitaker" 11/5/2004 11:22:44 AM >>> How many folks out there plan on doing the optional (for those already registered) Certification Maintenance Program? I think it's a good idea, and I plan on doing it, but I just wanted to know what the general feeling is. Will anyone require that of their employees? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: Friday, November 05, 2004 9:54 AM To: 'Jennifer MacDonald '; 'lpwenk@sbcglobal.net ' Cc: 'KMB1904@aol.com '; 'histonet@pathology.swmed.edu ' Subject: RE: [Histonet] CEU's Well, Florida sure does - I order mine through Anderson Continuing Education ; 3268 Ramos Circle / P.O.Box 276297 ; Sacramento, Ca 95827 Phone: 1-800-532-2332 / FAX 1-916-856-5066 They're really easy to use. You buy the book and they send a test booklet of multiple choice questions and a scantron sheet - which you can fax when you're done. You'll get a certificate in the mail within a week. Each course is mostly 24 CEUs with some exceptions that are 12 or 36. There's also one out of Miami, FL but I don't have the info on that. Give them a call!! Janet -----Original Message----- From: Jennifer MacDonald To: lpwenk@sbcglobal.net Cc: KMB1904@aol.com; histonet@pathology.swmed.edu Sent: 11/5/2004 1:46 AM Subject: Re: [Histonet] CEU's California has no requirements for Histotechnicians for certifica tion or continuing education. Jennifer MacDonald -----histonet-bounces@lists.utsouthwestern.= edu wrote: ----- To: , From: Sent by: histonet-bounces@lists.u= tsouthwestern.edu Date: 11/04/2004 06:15PM Subject: Re: [Histonet] CE= U's Multiple faceted question. I'll = try to answer it as best as I can. 1.There is NO national requiremen= t to have CEU to continue to work in histology (or cytology or med tech,= etc.). Most states have NO requirements (Florida and maybe Califo= rnia are exceptions). 2. Hospitals or your place of employment or ev= en you lab itself can set any requirement they want, including no requir= ement. 3. ASCP has a new requirement for all people passing a certif ication exam after Jan. 1, 2004. Previously, it was - once certified = certified for life. Now there is a Certification Maintenance Program. (= CMP) [1]= http://www.ascp.org/bor/cmp/index Now, if you want to maintain y= our certification status through ASCP, you must have 36 points of contin= uing education every 3 year, submit a form with this information along w= ith a renewal fee (currently $50). Documentation such as CEU forms are n= ot required, unless you are one of the randomly picked to be audited. Of= the 36 points, for HT/HTL, 1 point must be on safety, 2 points in your = area of specialty (i.e., histotechnology) and the rest of the points can= be in your area of specialty, management, education, or any related lab= area of interest. Points can be awarded for many types of CE, and e= ach type can be worth different number of points. For example: - 1= hour workshop =3D 1 point - 1 hour teleconference = 1 point - 1 se= mester hour of college =3D 15 points - competency assessment by your emp= loyer =3D 2 points per year - authoring an article in a peer reviewed jo= urnal (such as NSH Journal of Histotechnology or ASCP Laboratory Medicin= e) =3D 5 points. 4. Ways to get CE: Check in the ASCP CMP pac= ket (see above link, click on right side). Page 6 has lots of suggestion= s. - attending state meetings - attending national meetings - = in-services held at your institution - teleconferences (available throug= h NSH (301-262-6221) and ASCP (312-738-1336) - college classes (relat= ed to labs) - safety classes at your institution - competency assessm= ent - present at state and national meetings - write articles for his= totechnology journals - attend vendor workshops 36 points in 3 ye= ars really isn't a lot to have to get. And you don't have to spend a lot= of money. In-services are free - just make certain you have documentati= on (i.e. sign in sheet). Most places have you do safety quizzes - fire, = electrical, blood borne pathogens, etc. They would count. Peggy A. W= enk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Thursday, November 04, 2004 5:39 PM Subjec= t: [Histonet] CEU's > Does anyone know where you can ge= t CEU credits for histology? Are we > obligated to have so man= y a year? And if so, how many?? I have never seen this &g= t; information written anywhere. > > > ______ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F > Histonet mailing l= ist > Histonet@lists.utsouthwestern.edu > [2]http://li= sts.utsouthwestern.edu/mailman/listinfo/histonet ___ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F Histonet maili= ng list Histonet@lists.utsouthwestern.edu [3]http://lists.ut= southwestern.edu/mailman/listinfo/histonet <= /FONT> References 1. 3D"http://www.ascp.org/bor/cmp/index" 2. 3D"http://lis=/ 3. 3D"http://lists.uts=/ _______________________________________________ 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 _______________________________________________ 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 JNocito <@t> Pathreflab.com Fri Nov 5 11:09:54 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Dumbing down the practical??? In-Reply-To: Message-ID: I can do one better. Two techs came through my lab during a clinical rotation. They were so good that I hired them. Both took their practical at the same time using the same tissue from the same autopsy, used the same processor, microtome and stainer. I reviewed all their slides as did my medical director. One passed, one didn't. When I called ASCP to inquire how this could happen, I was told it would be $100 to have the slides reviewed. I told this person that I was the supervisor, not the applicant and I wanted to know how this could happen. I put my name in to become a reviewer, but never received a yes or no. Do I have some issues with ASCP, you bet. When my techs told me that they had to turn in only 9 slides, I was appalled. So I told the same old story "When I went to school, I had to walk 5 miles in waste high snow. When I did my HT, I had to use paraffin molds, process by hand and use a steel knife." Their response was "was that before cell phones and play stations?" They remember seeing a picture of a steel knife in a book once. Geez, am I getting old or what? Joe "The Toe" 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 TERRI BRAUD Sent: Thursday, November 04, 2004 3:52 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Dumbing down the practical??? I have a question for all those on the list who have taken the HT practical. Why do you think that the practical exam does not cover the material that it used to? Having taken mine in 1981, and then, later as a supervisor, having trained 8 OJT's to sucessful completion of their certification, I have observed that the practical is not near the test that it used to be. There is no CNS tissue, no decalcified tissue, no submission of both a special stain and a H&E from the same block. Just one slide, one block, one stain. Also how is the effect of fully automated special stainers vs prepared kits vs "homemade" solutions being taken into account? There isn't even an informational question pertaining to this so that the ASCP could begin to see the impact of the technology in our HT learning process? Will the technician that can successfully load an autostainer be as effective in troubleshooting a stain as one who has learned every step by hand? Will it even be necessary in the future of Histology? Just curious to hear some opinions - Terri _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Fri Nov 5 11:11:05 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] CEU's In-Reply-To: <000201c4c353$aec84980$3601a8c0@brownpathology.net> Message-ID: Bonnie, I'll do it if the same CEUs I take for my QIHC can be used for the CMP 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 Bonnie Whitaker Sent: Friday, November 05, 2004 10:23 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] CEU's How many folks out there plan on doing the optional (for those already registered) Certification Maintenance Program? I think it's a good idea, and I plan on doing it, but I just wanted to know what the general feeling is. Will anyone require that of their employees? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: Friday, November 05, 2004 9:54 AM To: 'Jennifer MacDonald '; 'lpwenk@sbcglobal.net ' Cc: 'KMB1904@aol.com '; 'histonet@pathology.swmed.edu ' Subject: RE: [Histonet] CEU's Well, Florida sure does - I order mine through Anderson Continuing Education ; 3268 Ramos Circle / P.O.Box 276297 ; Sacramento, Ca 95827 Phone: 1-800-532-2332 / FAX 1-916-856-5066 They're really easy to use. You buy the book and they send a test booklet of multiple choice questions and a scantron sheet - which you can fax when you're done. You'll get a certificate in the mail within a week. Each course is mostly 24 CEUs with some exceptions that are 12 or 36. There's also one out of Miami, FL but I don't have the info on that. Give them a call!! Janet -----Original Message----- From: Jennifer MacDonald To: lpwenk@sbcglobal.net Cc: KMB1904@aol.com; histonet@pathology.swmed.edu Sent: 11/5/2004 1:46 AM Subject: Re: [Histonet] CEU's California has no requirements for Histotechnicians for certifica tion or continuing education. Jennifer MacDonald -----histonet-bounces@lists.utsouthwestern.= edu wrote: ----- To: , From: Sent by: histonet-bounces@lists.u= tsouthwestern.edu Date: 11/04/2004 06:15PM Subject: Re: [Histonet] CE= U's Multiple faceted question. I'll = try to answer it as best as I can. 1.There is NO national requiremen= t to have CEU to continue to work in histology (or cytology or med tech,= etc.). Most states have NO requirements (Florida and maybe Califo= rnia are exceptions). 2. Hospitals or your place of employment or ev= en you lab itself can set any requirement they want, including no requir= ement. 3. ASCP has a new requirement for all people passing a certif ication exam after Jan. 1, 2004. Previously, it was - once certified = certified for life. Now there is a Certification Maintenance Program. (= CMP) [1]= http://www.ascp.org/bor/cmp/index Now, if you want to maintain y= our certification status through ASCP, you must have 36 points of contin= uing education every 3 year, submit a form with this information along w= ith a renewal fee (currently $50). Documentation such as CEU forms are n= ot required, unless you are one of the randomly picked to be audited. Of= the 36 points, for HT/HTL, 1 point must be on safety, 2 points in your = area of specialty (i.e., histotechnology) and the rest of the points can= be in your area of specialty, management, education, or any related lab= area of interest. Points can be awarded for many types of CE, and e= ach type can be worth different number of points. For example: - 1= hour workshop =3D 1 point - 1 hour teleconference = 1 point - 1 se= mester hour of college =3D 15 points - competency assessment by your emp= loyer =3D 2 points per year - authoring an article in a peer reviewed jo= urnal (such as NSH Journal of Histotechnology or ASCP Laboratory Medicin= e) =3D 5 points. 4. Ways to get CE: Check in the ASCP CMP pac= ket (see above link, click on right side). Page 6 has lots of suggestion= s. - attending state meetings - attending national meetings - = in-services held at your institution - teleconferences (available throug= h NSH (301-262-6221) and ASCP (312-738-1336) - college classes (relat= ed to labs) - safety classes at your institution - competency assessm= ent - present at state and national meetings - write articles for his= totechnology journals - attend vendor workshops 36 points in 3 ye= ars really isn't a lot to have to get. And you don't have to spend a lot= of money. In-services are free - just make certain you have documentati= on (i.e. sign in sheet). Most places have you do safety quizzes - fire, = electrical, blood borne pathogens, etc. They would count. Peggy A. W= enk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Thursday, November 04, 2004 5:39 PM Subjec= t: [Histonet] CEU's > Does anyone know where you can ge= t CEU credits for histology? Are we > obligated to have so man= y a year? And if so, how many?? I have never seen this &g= t; information written anywhere. > > > ______ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F > Histonet mailing l= ist > Histonet@lists.utsouthwestern.edu > [2]http://li= sts.utsouthwestern.edu/mailman/listinfo/histonet ___ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F Histonet maili= ng list Histonet@lists.utsouthwestern.edu [3]http://lists.ut= southwestern.edu/mailman/listinfo/histonet <= /FONT> References 1. 3D"http://www.ascp.org/bor/cmp/index" 2. 3D"http://lis=/ 3. 3D"http://lists.uts=/ _______________________________________________ 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 From JQB7 <@t> CDC.GOV Fri Nov 5 11:12:26 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] CEU's Message-ID: I think every single histotech out there regardless of when they were certified should be required to do this. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: Friday, November 05, 2004 12:07 PM To: 'Rebecca Barnhart'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] CEU's It's a good idea, especially if you go state to state, or just need an update. I've been in several states and heard talk of starting the CEU program in all these locals. AND I would not be surprised if us "oldsters" won't be required to do it in the future! I've taken a couple of courses that were good updates - the Histology field is no longer stagnant as it seemed to be from 1940-1970. -----Original Message----- From: Rebecca Barnhart [mailto:rbarnhart@summithealth.org] Sent: Friday, November 05, 2004 11:49 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] CEU's I think it is a good idea and also am planning on doing it. I would not be opposed if my employeer requires it for the entire lab staff (MT, MLT, CT and the 1 phelb that we have) >>> "Bonnie Whitaker" 11/5/2004 11:22:44 AM >>> How many folks out there plan on doing the optional (for those already registered) Certification Maintenance Program? I think it's a good idea, and I plan on doing it, but I just wanted to know what the general feeling is. Will anyone require that of their employees? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: Friday, November 05, 2004 9:54 AM To: 'Jennifer MacDonald '; 'lpwenk@sbcglobal.net ' Cc: 'KMB1904@aol.com '; 'histonet@pathology.swmed.edu ' Subject: RE: [Histonet] CEU's Well, Florida sure does - I order mine through Anderson Continuing Education ; 3268 Ramos Circle / P.O.Box 276297 ; Sacramento, Ca 95827 Phone: 1-800-532-2332 / FAX 1-916-856-5066 They're really easy to use. You buy the book and they send a test booklet of multiple choice questions and a scantron sheet - which you can fax when you're done. You'll get a certificate in the mail within a week. Each course is mostly 24 CEUs with some exceptions that are 12 or 36. There's also one out of Miami, FL but I don't have the info on that. Give them a call!! Janet -----Original Message----- From: Jennifer MacDonald To: lpwenk@sbcglobal.net Cc: KMB1904@aol.com; histonet@pathology.swmed.edu Sent: 11/5/2004 1:46 AM Subject: Re: [Histonet] CEU's California has no requirements for Histotechnicians for certifica tion or continuing education. Jennifer MacDonald -----histonet-bounces@lists.utsouthwestern.= edu wrote: ----- To: , From: Sent by: histonet-bounces@lists.u= tsouthwestern.edu Date: 11/04/2004 06:15PM Subject: Re: [Histonet] CE= U's Multiple faceted question. I'll = try to answer it as best as I can. 1.There is NO national requiremen= t to have CEU to continue to work in histology (or cytology or med tech,= etc.). Most states have NO requirements (Florida and maybe Califo= rnia are exceptions). 2. Hospitals or your place of employment or ev= en you lab itself can set any requirement they want, including no requir= ement. 3. ASCP has a new requirement for all people passing a certif ication exam after Jan. 1, 2004. Previously, it was - once certified = certified for life. Now there is a Certification Maintenance Program. (= CMP) [1]= http://www.ascp.org/bor/cmp/index Now, if you want to maintain y= our certification status through ASCP, you must have 36 points of contin= uing education every 3 year, submit a form with this information along w= ith a renewal fee (currently $50). Documentation such as CEU forms are n= ot required, unless you are one of the randomly picked to be audited. Of= the 36 points, for HT/HTL, 1 point must be on safety, 2 points in your = area of specialty (i.e., histotechnology) and the rest of the points can= be in your area of specialty, management, education, or any related lab= area of interest. Points can be awarded for many types of CE, and e= ach type can be worth different number of points. For example: - 1= hour workshop =3D 1 point - 1 hour teleconference = 1 point - 1 se= mester hour of college =3D 15 points - competency assessment by your emp= loyer =3D 2 points per year - authoring an article in a peer reviewed jo= urnal (such as NSH Journal of Histotechnology or ASCP Laboratory Medicin= e) =3D 5 points. 4. Ways to get CE: Check in the ASCP CMP pac= ket (see above link, click on right side). Page 6 has lots of suggestion= s. - attending state meetings - attending national meetings - = in-services held at your institution - teleconferences (available throug= h NSH (301-262-6221) and ASCP (312-738-1336) - college classes (relat= ed to labs) - safety classes at your institution - competency assessm= ent - present at state and national meetings - write articles for his= totechnology journals - attend vendor workshops 36 points in 3 ye= ars really isn't a lot to have to get. And you don't have to spend a lot= of money. In-services are free - just make certain you have documentati= on (i.e. sign in sheet). Most places have you do safety quizzes - fire, = electrical, blood borne pathogens, etc. They would count. Peggy A. W= enk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Thursday, November 04, 2004 5:39 PM Subjec= t: [Histonet] CEU's > Does anyone know where you can ge= t CEU credits for histology? Are we > obligated to have so man= y a year? And if so, how many?? I have never seen this &g= t; information written anywhere. > > > ______ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F > Histonet mailing l= ist > Histonet@lists.utsouthwestern.edu > [2]http://li= sts.utsouthwestern.edu/mailman/listinfo/histonet ___ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F Histonet maili= ng list Histonet@lists.utsouthwestern.edu [3]http://lists.ut= southwestern.edu/mailman/listinfo/histonet <= /FONT> References 1. 3D"http://www.ascp.org/bor/cmp/index" 2. 3D"http://lis=/ 3. 3D"http://lists.uts=/ _______________________________________________ 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 _______________________________________________ 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 GoodwinD <@t> pahosp.com Fri Nov 5 11:20:26 2004 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] question for listserver screener Message-ID: <992899E9EC268548AB8DDE246AF88473055F4B9E@PAHEX01.uphs.upenn.edu> Do you allow postings for job openings, or for membership recruitment for state societies? Thanks, Diana Goodwin Vice President, NewJersey Society for Histotechnology Pennsylvania Hospital 800 Spruce St., Preston 655-C Philadelphia, PA 19107 ph:215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com From GoodwinD <@t> pahosp.com Fri Nov 5 11:21:55 2004 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] question for listserver screener Message-ID: <992899E9EC268548AB8DDE246AF88473055F4B9F@PAHEX01.uphs.upenn.edu> Do you allow postings for job openings, or for membership recruitment for state societies? Thanks, Diana Goodwin Vice President, NewJersey Society for Histotechnology Pennsylvania Hospital 800 Spruce St., Preston 655-C Philadelphia, PA 19107 ph:215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com From Lynne.Bell <@t> hitchcock.org Fri Nov 5 11:22:35 2004 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] CEU's Message-ID: I also plan on doing the CMP and we have talked in our leadership group of requiring it for all registered laboratory personnel. I think it is a great idea to keep all of us current. Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 From froyer <@t> bitstream.net Fri Nov 5 11:36:33 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] a much easier chemistry question In-Reply-To: <418AC26B.98C6BB84@uwo.ca> References: <418AC26B.98C6BB84@uwo.ca> Message-ID: <418BBA21.7040605@bitstream.net> Years ago, Scientific Products made a lanolin-based cream stain remover that worked great in removing biological stains. It worked like magic on stain on hands and lab surfaces. One day, out of frustration, I tried it on my white lab coat (stained with methylene blue) and it worked fine. I rubbed the cream into the stain, and then washed to coat in the washing machine... stain all gone! I check a recent (1997) Allegiance catalog, and they still showed this product. Called "Dye-Sol" and the catalog number was C6345. All the S/P products are now carried by Cardinal so you could contact them to see if they still carry it. They should be able to cross-reference the old Allegiance cat. no. Good Luck! ~ Ford Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN John Kiernan wrote: >Your unsuccessful efforts were all acidic, >neutral or hydrophobic. Trypan and Evans blue are >anionic dyes with large, very hydrophilic molecules. >Alkaline solutions extract anionic dyes, and urea >(high concentration) can disrupt the non-ionic, >non-covalent bonding that holds large hydrophilic >dye molecules to substrates such as cellulose (cotton). > >Try a strong urea solution (8M=48%) made alkaline >with borax or washing soda. Geoff McAuliffe's >simpler suggestion of soap and water may work, >because soap is alkaline. > >Keep us informed. All who use dyes need this sort >of information. (I have a grey tie with alcian >blue spots dating from the 1960s; wouldn't want to >remove them now, though I'm sure it would be >impossible without dissolving the fabric.) > >John Kiernan >London, Canada >------------------------------------- >Jack England wrote: > > >>Aloha all, >> >>Here's a much more fun chemistry question for anyone that wants to bite: >>what is the best method to remove trypan/evans blue from cotton fabric? One >>of my colleagues got some on her pants the other day and asked what the best >>way to get it out was. I did not know, so I put some on cotton gauze, and >>tried: >> >>--acid alcohol...faded the stain a little, but didn't get rid of it >>--DI water...no change >>--absolute ethanol...no change >>--xylene...no change >>--ethanol/non-ionic detergent/DI water rinse...no change >>--citrus oil clearant...no change >> >>Given that her fabric is tan (and thus not white, and thus likely not >>bleach-able), can anyone out there in histo-land suggest a way of >>de-staining trypan-blue-stained fabric? I've been wondering about this for a >>while now and figured I'd pass it around. >> >>--Many thanks and aloha to all, >>Jack England >>Tissue Genesis, Inc. >>http://www.tissuegenesis.com >> >>_________________________________________________________________ >>Express yourself instantly with MSN Messenger! Download today - it's FREE! >>hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ >> >>_______________________________________________ >>Histonet mailing list >>Histonet@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 JEllin <@t> yumaregional.org Fri Nov 5 11:33:51 2004 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] CEU's Message-ID: I also plan on doing my CEU's Jesus Ellin >>> "Bell, Lynne" 11/05/04 10:22AM >>> I also plan on doing the CMP and we have talked in our leadership group of requiring it for all registered laboratory personnel. I think it is a great idea to keep all of us current. 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 JEllin <@t> yumaregional.org Fri Nov 5 11:33:51 2004 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] CEU's Message-ID: I also plan on doing my CEU's Jesus Ellin >>> "Bell, Lynne" 11/05/04 10:22AM >>> I also plan on doing the CMP and we have talked in our leadership group of requiring it for all registered laboratory personnel. I think it is a great idea to keep all of us current. 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 vbaker60 <@t> yahoo.com Fri Nov 5 11:50:41 2004 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Contact information for Belair Instruments Message-ID: <20041105175041.67866.qmail@web50109.mail.yahoo.com> Hi Everyone, Does anyone have the website or e-mail contact information for Belair Instruments? I'll even take a phone number!! I had all this information when I was still at my former place of work, but I haven't unpacked ALL my many, many boxes yet. Thanks in advance and have a nice weekend. Vikki Baker Mt. Sinai School of Medicine 1425 Madison Ave NY, NY 10029 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From MAUGER <@t> email.chop.edu Fri Nov 5 11:56:08 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] CEU's Message-ID: Bonnie, I agree with Joe. I do it for the QIHC also. I wish my co workers had to do it for histo. Jo >>> "Joe Nocito" 11/05/04 12:11PM >>> Bonnie, I'll do it if the same CEUs I take for my QIHC can be used for the CMP 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 Bonnie Whitaker Sent: Friday, November 05, 2004 10:23 AM To: histonet@pathology.swmed.edu Subject: RE: [Histonet] CEU's How many folks out there plan on doing the optional (for those already registered) Certification Maintenance Program? I think it's a good idea, and I plan on doing it, but I just wanted to know what the general feeling is. Will anyone require that of their employees? Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: Friday, November 05, 2004 9:54 AM To: 'Jennifer MacDonald '; 'lpwenk@sbcglobal.net ' Cc: 'KMB1904@aol.com '; 'histonet@pathology.swmed.edu ' Subject: RE: [Histonet] CEU's Well, Florida sure does - I order mine through Anderson Continuing Education ; 3268 Ramos Circle / P.O.Box 276297 ; Sacramento, Ca 95827 Phone: 1-800-532-2332 / FAX 1-916-856-5066 They're really easy to use. You buy the book and they send a test booklet of multiple choice questions and a scantron sheet - which you can fax when you're done. You'll get a certificate in the mail within a week. Each course is mostly 24 CEUs with some exceptions that are 12 or 36. There's also one out of Miami, FL but I don't have the info on that. Give them a call!! Janet -----Original Message----- From: Jennifer MacDonald To: lpwenk@sbcglobal.net Cc: KMB1904@aol.com; histonet@pathology.swmed.edu Sent: 11/5/2004 1:46 AM Subject: Re: [Histonet] CEU's California has no requirements for Histotechnicians for certifica tion or continuing education. Jennifer MacDonald -----histonet-bounces@lists.utsouthwestern.= edu wrote: ----- To: , From: Sent by: histonet-bounces@lists.u= tsouthwestern.edu Date: 11/04/2004 06:15PM Subject: Re: [Histonet] CE= U's Multiple faceted question. I'll = try to answer it as best as I can. 1.There is NO national requiremen= t to have CEU to continue to work in histology (or cytology or med tech,= etc.). Most states have NO requirements (Florida and maybe Califo= rnia are exceptions). 2. Hospitals or your place of employment or ev= en you lab itself can set any requirement they want, including no requir= ement. 3. ASCP has a new requirement for all people passing a certif ication exam after Jan. 1, 2004. Previously, it was - once certified = certified for life. Now there is a Certification Maintenance Program. (= CMP) [1]= http://www.ascp.org/bor/cmp/index Now, if you want to maintain y= our certification status through ASCP, you must have 36 points of contin= uing education every 3 year, submit a form with this information along w= ith a renewal fee (currently $50). Documentation such as CEU forms are n= ot required, unless you are one of the randomly picked to be audited. Of= the 36 points, for HT/HTL, 1 point must be on safety, 2 points in your = area of specialty (i.e., histotechnology) and the rest of the points can= be in your area of specialty, management, education, or any related lab= area of interest. Points can be awarded for many types of CE, and e= ach type can be worth different number of points. For example: - 1= hour workshop =3D 1 point - 1 hour teleconference = 1 point - 1 se= mester hour of college =3D 15 points - competency assessment by your emp= loyer =3D 2 points per year - authoring an article in a peer reviewed jo= urnal (such as NSH Journal of Histotechnology or ASCP Laboratory Medicin= e) =3D 5 points. 4. Ways to get CE: Check in the ASCP CMP pac= ket (see above link, click on right side). Page 6 has lots of suggestion= s. - attending state meetings - attending national meetings - = in-services held at your institution - teleconferences (available throug= h NSH (301-262-6221) and ASCP (312-738-1336) - college classes (relat= ed to labs) - safety classes at your institution - competency assessm= ent - present at state and national meetings - write articles for his= totechnology journals - attend vendor workshops 36 points in 3 ye= ars really isn't a lot to have to get. And you don't have to spend a lot= of money. In-services are free - just make certain you have documentati= on (i.e. sign in sheet). Most places have you do safety quizzes - fire, = electrical, blood borne pathogens, etc. They would count. Peggy A. W= enk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: To: Sent: Thursday, November 04, 2004 5:39 PM Subjec= t: [Histonet] CEU's > Does anyone know where you can ge= t CEU credits for histology? Are we > obligated to have so man= y a year? And if so, how many?? I have never seen this &g= t; information written anywhere. > > > ______ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F > Histonet mailing l= ist > Histonet@lists.utsouthwestern.edu > [2]http://li= sts.utsouthwestern.edu/mailman/listinfo/histonet ___ ______________________ 5F__= =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F Histonet maili= ng list Histonet@lists.utsouthwestern.edu [3]http://lists.ut= southwestern.edu/mailman/listinfo/histonet <= /FONT> References 1. 3D"http://www.ascp.org/bor/cmp/index" 2. 3D"http://lis=/ 3. 3D"http://lists.uts=/ _______________________________________________ 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From japoteete <@t> saintfrancis.com Fri Nov 5 12:06:52 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Dumbing down the practical??? Message-ID: WASTE high snow! What a horrible experience! Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Lab Saint Francis Hospital Tulsa, OK > -----Original Message----- > From: Joe Nocito [SMTP:JNocito@Pathreflab.com] > Sent: Friday, November 05, 2004 11:10 AM > To: TERRI BRAUD; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Dumbing down the practical??? > > I can do one better. > Two techs came through my lab during a clinical rotation. They were so > good > that I hired them. Both took their practical at the same time using the > same > tissue from the same autopsy, used the same processor, microtome and > stainer. I reviewed all their slides as did my medical director. One > passed, > one didn't. When I called ASCP to inquire how this could happen, I was > told > it would be $100 to have the slides reviewed. I told this person that I > was > the supervisor, not the applicant and I wanted to know how this could > happen. > I put my name in to become a reviewer, but never received a yes or > no. > Do I have some issues with ASCP, you bet. When my techs told me that they > had to turn in only 9 slides, I was appalled. So I told the same old story > "When I went to school, I had to walk 5 miles in waste high snow. When I > did > my HT, I had to use paraffin molds, process by hand and use a steel > knife." > Their response was "was that before cell phones and play stations?" They > remember seeing a picture of a steel knife in a book once. Geez, am I > getting old or what? > > Joe "The Toe" 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 TERRI > BRAUD > Sent: Thursday, November 04, 2004 3:52 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Dumbing down the practical??? > > > I have a question for all those on the list who have taken the HT > practical. > Why do you think that the practical exam does not cover the material > that > it used to? Having taken mine in 1981, and then, later as a supervisor, > having trained 8 OJT's to sucessful completion of their certification, I > have observed that the practical is not near the test that it used to be. > There is no CNS tissue, no decalcified tissue, no submission of both a > special stain and a H&E from the same block. Just one slide, one block, > one > stain. Also how is the effect of fully automated special stainers vs > prepared kits vs "homemade" solutions being taken into account? There > isn't > even an informational question pertaining to this so that the ASCP could > begin to see the impact of the technology in our HT learning process? > Will > the technician that can successfully load an autostainer be as effective > in > troubleshooting a stain as one who has learned every step by hand? Will it > even be necessary in the future of Histology? Just curious to hear some > opinions - Terri > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From Jackie.O'Connor <@t> abbott.com Fri Nov 5 12:32:01 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] a much easier chemistry question Message-ID: Hasn't anyone tried Oxy Clean? I even use it on my white show dogs (then a little bit of Downy in the rinse water to make their coats soft). They smell good, too. Jackie O' Ford Royer Sent by: histonet-bounces@lists.utsouthwestern.edu 11/05/2004 11:36 AM To: Histonet@lists.utsouthwestern.edu cc: Subject: Re: [Histonet] a much easier chemistry question Years ago, Scientific Products made a lanolin-based cream stain remover that worked great in removing biological stains. It worked like magic on stain on hands and lab surfaces. One day, out of frustration, I tried it on my white lab coat (stained with methylene blue) and it worked fine. I rubbed the cream into the stain, and then washed to coat in the washing machine... stain all gone! I check a recent (1997) Allegiance catalog, and they still showed this product. Called "Dye-Sol" and the catalog number was C6345. All the S/P products are now carried by Cardinal so you could contact them to see if they still carry it. They should be able to cross-reference the old Allegiance cat. no. Good Luck! ~ Ford Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN John Kiernan wrote: >Your unsuccessful efforts were all acidic, >neutral or hydrophobic. Trypan and Evans blue are >anionic dyes with large, very hydrophilic molecules. >Alkaline solutions extract anionic dyes, and urea >(high concentration) can disrupt the non-ionic, >non-covalent bonding that holds large hydrophilic >dye molecules to substrates such as cellulose (cotton). > >Try a strong urea solution (8M=48%) made alkaline >with borax or washing soda. Geoff McAuliffe's >simpler suggestion of soap and water may work, >because soap is alkaline. > >Keep us informed. All who use dyes need this sort >of information. (I have a grey tie with alcian >blue spots dating from the 1960s; wouldn't want to >remove them now, though I'm sure it would be >impossible without dissolving the fabric.) > >John Kiernan >London, Canada >------------------------------------- >Jack England wrote: > > >>Aloha all, >> >>Here's a much more fun chemistry question for anyone that wants to bite: >>what is the best method to remove trypan/evans blue from cotton fabric? One >>of my colleagues got some on her pants the other day and asked what the best >>way to get it out was. I did not know, so I put some on cotton gauze, and >>tried: >> >>--acid alcohol...faded the stain a little, but didn't get rid of it >>--DI water...no change >>--absolute ethanol...no change >>--xylene...no change >>--ethanol/non-ionic detergent/DI water rinse...no change >>--citrus oil clearant...no change >> >>Given that her fabric is tan (and thus not white, and thus likely not >>bleach-able), can anyone out there in histo-land suggest a way of >>de-staining trypan-blue-stained fabric? I've been wondering about this for a >>while now and figured I'd pass it around. >> >>--Many thanks and aloha to all, >>Jack England >>Tissue Genesis, Inc. >>http://www.tissuegenesis.com >> >>_________________________________________________________________ >>Express yourself instantly with MSN Messenger! Download today - it's FREE! >>hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ >> >>_______________________________________________ >>Histonet mailing list >>Histonet@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 Nov 5 12:37:37 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] a much easier chemistry question Message-ID: <83AACDB0810528418AA106F9AE9B7F7E5074AF@sjhaexc02.sjha.org> And that would be ERADO-SOL C6347 It really does work! 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 Ford Royer Sent: Friday, November 05, 2004 12:37 PM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] a much easier chemistry question Years ago, Scientific Products made a lanolin-based cream stain remover that worked great in removing biological stains. It worked like magic on stain on hands and lab surfaces. One day, out of frustration, I tried it on my white lab coat (stained with methylene blue) and it worked fine. I rubbed the cream into the stain, and then washed to coat in the washing machine... stain all gone! I check a recent (1997) Allegiance catalog, and they still showed this product. Called "Dye-Sol" and the catalog number was C6345. All the S/P products are now carried by Cardinal so you could contact them to see if they still carry it. They should be able to cross-reference the old Allegiance cat. no. Good Luck! ~ Ford Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN John Kiernan wrote: >Your unsuccessful efforts were all acidic, >neutral or hydrophobic. Trypan and Evans blue are >anionic dyes with large, very hydrophilic molecules. >Alkaline solutions extract anionic dyes, and urea >(high concentration) can disrupt the non-ionic, >non-covalent bonding that holds large hydrophilic >dye molecules to substrates such as cellulose (cotton). > >Try a strong urea solution (8M=48%) made alkaline >with borax or washing soda. Geoff McAuliffe's >simpler suggestion of soap and water may work, >because soap is alkaline. > >Keep us informed. All who use dyes need this sort >of information. (I have a grey tie with alcian >blue spots dating from the 1960s; wouldn't want to >remove them now, though I'm sure it would be >impossible without dissolving the fabric.) > >John Kiernan >London, Canada >------------------------------------- >Jack England wrote: > > >>Aloha all, >> >>Here's a much more fun chemistry question for anyone that wants to bite: >>what is the best method to remove trypan/evans blue from cotton fabric? One >>of my colleagues got some on her pants the other day and asked what the best >>way to get it out was. I did not know, so I put some on cotton gauze, and >>tried: >> >>--acid alcohol...faded the stain a little, but didn't get rid of it >>--DI water...no change >>--absolute ethanol...no change >>--xylene...no change >>--ethanol/non-ionic detergent/DI water rinse...no change >>--citrus oil clearant...no change >> >>Given that her fabric is tan (and thus not white, and thus likely not >>bleach-able), can anyone out there in histo-land suggest a way of >>de-staining trypan-blue-stained fabric? I've been wondering about this for a >>while now and figured I'd pass it around. >> >>--Many thanks and aloha to all, >>Jack England >>Tissue Genesis, Inc. >>http://www.tissuegenesis.com >> > 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 Fri Nov 5 12:40:22 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Re: Waist high snow Message-ID: My kids had the benefit of living and going to school in Hawaii. They are already using the following phrase: "Why, when I was a kid, I used to have to take a boat across shark infested Pearl Harbor in the dead of winter to get to school". They DID. We lived on a base on the leeward side of Pearl Harbor, and the Navy provided a 200 man shuttle/launch to get to the Windward side for school kids and military personnel to get to work at Pearl Harbor and Hickam Air Force Base. So, for all you parents who worry when you put your kids on the school BUS every day - Pearl Harbor IS infested with hammer head sharks. I'll never forget yelling "Hurry up! You'll miss the school boat!" "Poteete, Jacquie A." Sent by: histonet-bounces@lists.utsouthwestern.edu 11/05/2004 12:06 PM To: "'Joe Nocito'" , TERRI BRAUD , histonet@lists.utsouthwestern.edu cc: Subject: RE: [Histonet] Dumbing down the practical??? WASTE high snow! What a horrible experience! Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Lab Saint Francis Hospital Tulsa, OK > -----Original Message----- > From: Joe Nocito [SMTP:JNocito@Pathreflab.com] > Sent: Friday, November 05, 2004 11:10 AM > To: TERRI BRAUD; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Dumbing down the practical??? > > I can do one better. > Two techs came through my lab during a clinical rotation. They were so > good > that I hired them. Both took their practical at the same time using the > same > tissue from the same autopsy, used the same processor, microtome and > stainer. I reviewed all their slides as did my medical director. One > passed, > one didn't. When I called ASCP to inquire how this could happen, I was > told > it would be $100 to have the slides reviewed. I told this person that I > was > the supervisor, not the applicant and I wanted to know how this could > happen. > I put my name in to become a reviewer, but never received a yes or > no. > Do I have some issues with ASCP, you bet. When my techs told me that they > had to turn in only 9 slides, I was appalled. So I told the same old story > "When I went to school, I had to walk 5 miles in waste high snow. When I > did > my HT, I had to use paraffin molds, process by hand and use a steel > knife." > Their response was "was that before cell phones and play stations?" They > remember seeing a picture of a steel knife in a book once. Geez, am I > getting old or what? > > Joe "The Toe" 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 TERRI > BRAUD > Sent: Thursday, November 04, 2004 3:52 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Dumbing down the practical??? > > > I have a question for all those on the list who have taken the HT > practical. > Why do you think that the practical exam does not cover the material > that > it used to? Having taken mine in 1981, and then, later as a supervisor, > having trained 8 OJT's to sucessful completion of their certification, I > have observed that the practical is not near the test that it used to be. > There is no CNS tissue, no decalcified tissue, no submission of both a > special stain and a H&E from the same block. Just one slide, one block, > one > stain. Also how is the effect of fully automated special stainers vs > prepared kits vs "homemade" solutions being taken into account? There > isn't > even an informational question pertaining to this so that the ASCP could > begin to see the impact of the technology in our HT learning process? > Will > the technician that can successfully load an autostainer be as effective > in > troubleshooting a stain as one who has learned every step by hand? Will it > even be necessary in the future of Histology? Just curious to hear some > opinions - Terri > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri Nov 5 12:48:42 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] IHC on cell cultures Message-ID: Can anyone help me with a protocol for preparing cell culture samples for IHC? I'm particularly interested in looking at CD45. I'm not quite sure of the best way to prepare the cells. I can do cytospins - and I'll bet I have to fix the cells prior to cytospinning them. If anyone has a protocol they'd like to share for preparing good slides from live cells, I'd appreciate it. I'd like to keep the option of using cytospins or smears as well as making a cell block. I'm experienced with making these preparations from body fluids, but not cell cultures. Thanks. Jackie O'. From Don.Birgerson <@t> leica-microsystems.com Fri Nov 5 12:53:46 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Contact information for Belair Instruments Message-ID: Hi Vikki, The number for Belair Instruments is 973-912-8900. If you have further questions, please call. Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 Victoria Baker To: HistoNet Server Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Contact information for Belair Instruments western.edu 11/05/2004 11:50 AM Hi Everyone, Does anyone have the website or e-mail contact information for Belair Instruments? I'll even take a phone number!! I had all this information when I was still at my former place of work, but I haven't unpacked ALL my many, many boxes yet. Thanks in advance and have a nice weekend. Vikki Baker Mt. Sinai School of Medicine 1425 Madison Ave NY, NY 10029 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From scoop <@t> mail.nih.gov Fri Nov 5 13:08:59 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] IHC on cell cultures In-Reply-To: References: Message-ID: If the cells grow in a monolayer, just throw in a few cover slips (you may or may not have to treat the cover slips to get the cells to stick, depending on the cells) and then fix with whatever you like (4% PF in NBS, ice cold methanol, acetone, etc.). If the cells grow in spinner culture, I think the best method is to do a cytospin. Sharon >Can anyone help me with a protocol for preparing cell culture samples for >IHC? I'm particularly interested in looking at CD45. I'm not quite sure >of the best way to prepare the cells. I can do cytospins - and I'll bet I >have to fix the cells prior to cytospinning them. If anyone has a >protocol they'd like to share for preparing good slides from live cells, >I'd appreciate it. I'd like to keep the option of using cytospins or >smears as well as making a cell block. I'm experienced with making these >preparations from body fluids, but not cell cultures. Thanks. > >Jackie O'. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From ohenry <@t> dfw.net Fri Nov 5 13:12:34 2004 From: ohenry <@t> dfw.net (Susan Owens) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Re: Dumbing down the practical. Message-ID: <004a01c4c36b$6a2a71a0$e0dd3040@Nationwide.net> >1. In all those years, there has never been a time that both the H&E and a >special stain has been requested on the same tissue. That would be double >penalizing someone if they had a bad tissue. Well I don't agree. It's been a few years, but I seem to remember that I did special stains and H&E on select blocks when I took the exam(in 1964).. And this thing about a 'double penalizing someone' for a bad tissue...Well they should not of used 'bad tissue' to begin with. And if they didn't know the tissue was 'bad' then they shouldn't of been taking the exam. I believe there are too many new faces in histoland that think they know it all and in truth they don't have a clue. ex: 1. A year or two ago I asked someone to make up a % solution of something(I don't remember what)..I was told they didn't know how. I asked 'how could you not know,you have your HT?' They answered "that they didn't need to know, where they worked before they purchased everything." 2. This year someone I know was taking the practical. According to them they were having a problem getting one of the special stains to work. SO, after several tries, they just put it on the auto stainer then sent it in with all the other slides. What did this person learn? NOTHING And what did ASCP learn? NOTHING (Do they understand they are handing out HT's to some people who don't know how to do the work?) These are just two examples, I could list allot more....But my blood pressure would go wild. Just keep smiling. Susan From froyer <@t> bitstream.net Fri Nov 5 13:26:14 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Dumbing down the practical???] Message-ID: <418BD3D6.1000305@bitstream.net> ...and I bet it was up-hill BOTH WAYS!!! ;-) ~ Ford ps: You're dating yourself Joe... but then my Medical Technology class was the last class that was required to go through a rotation in Histology, and we had 25 Histo/Cyto questions included on that year's M.T. Registry.... and they were HARD questions! Maybe that's why they dropped Histology from the Med Tech requirements... it was to hard for us . ;-) Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN Poteete, Jacquie A. wrote: >WASTE high snow! What a horrible experience! > >Jacquie Poteete MT(ASCP)QIHC >Lead Technologist, IHC Lab >Saint Francis Hospital >Tulsa, OK > > > >>-----Original Message----- >>From: Joe Nocito [SMTP:JNocito@Pathreflab.com] >>Sent: Friday, November 05, 2004 11:10 AM >>To: TERRI BRAUD; histonet@lists.utsouthwestern.edu >>Subject: RE: [Histonet] Dumbing down the practical??? >> >>I can do one better. >>Two techs came through my lab during a clinical rotation. They were so >>good >>that I hired them. Both took their practical at the same time using the >>same >>tissue from the same autopsy, used the same processor, microtome and >>stainer. I reviewed all their slides as did my medical director. One >>passed, >>one didn't. When I called ASCP to inquire how this could happen, I was >>told >>it would be $100 to have the slides reviewed. I told this person that I >>was >>the supervisor, not the applicant and I wanted to know how this could >>happen. >> I put my name in to become a reviewer, but never received a yes or >>no. >>Do I have some issues with ASCP, you bet. When my techs told me that they >>had to turn in only 9 slides, I was appalled. So I told the same old story >>"When I went to school, I had to walk 5 miles in waste high snow. When I >>did >>my HT, I had to use paraffin molds, process by hand and use a steel >>knife." >>Their response was "was that before cell phones and play stations?" They >>remember seeing a picture of a steel knife in a book once. Geez, am I >>getting old or what? >> >>Joe "The Toe" 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 TERRI >>BRAUD >>Sent: Thursday, November 04, 2004 3:52 PM >>To: histonet@lists.utsouthwestern.edu >>Subject: [Histonet] Dumbing down the practical??? >> >> >>I have a question for all those on the list who have taken the HT >>practical. >> Why do you think that the practical exam does not cover the material >>that >>it used to? Having taken mine in 1981, and then, later as a supervisor, >>having trained 8 OJT's to sucessful completion of their certification, I >>have observed that the practical is not near the test that it used to be. >>There is no CNS tissue, no decalcified tissue, no submission of both a >>special stain and a H&E from the same block. Just one slide, one block, >>one >>stain. Also how is the effect of fully automated special stainers vs >>prepared kits vs "homemade" solutions being taken into account? There >>isn't >>even an informational question pertaining to this so that the ASCP could >>begin to see the impact of the technology in our HT learning process? >>Will >>the technician that can successfully load an autostainer be as effective >>in >>troubleshooting a stain as one who has learned every step by hand? Will it >>even be necessary in the future of Histology? Just curious to hear some >>opinions - Terri >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> >> >> >********* Email Confidentiality Statement ********* >Visit http://www.saintfrancis.com/emailconf.asp > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From ander093 <@t> tc.umn.edu Fri Nov 5 13:52:45 2004 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Re: Dumbing down the practical. In-Reply-To: <004a01c4c36b$6a2a71a0$e0dd3040@Nationwide.net> References: <004a01c4c36b$6a2a71a0$e0dd3040@Nationwide.net> Message-ID: <6.1.2.0.0.20041105134651.028b2780@ander093.email.umn.edu> I also remember being required to do specials and H&E's on the same blocks. If it turned out to be a bad section/sections--or bad tissue--it was back to the drawing board!! Recuts or new tissue were then in order!! AND, not to date myself, but everything was done by hand with the exception of processing!! I too,believe that it is important have that "hands on" know how for troubleshooting purposes. JMHO. LuAnn At 01:12 PM 11/5/04, Susan Owens wrote: > >1. In all those years, there has never been a time that both the H&E and a > >special stain has been requested on the same tissue. That would be double > >penalizing someone if they had a bad tissue. > > > >Well I don't agree. It's been a few years, but I seem to remember that I did >special stains and H&E on select blocks when I took the exam(in 1964).. And >this thing about a 'double penalizing someone' for a bad tissue...Well they >should not of used 'bad tissue' to begin with. And if they didn't know the >tissue was 'bad' then they shouldn't of been taking the exam. >I believe there are too many new faces in histoland that think they know it >all and in truth they don't have a clue. > >ex: 1. A year or two ago I asked someone to make up a % solution of >something(I don't remember what)..I was told they didn't know how. I asked >'how could you not know,you have your HT?' They answered "that they didn't >need to know, where they worked before they purchased everything." > 2. This year someone I know was taking the practical. According to >them they were having a problem getting one of the special stains to work. >SO, after several tries, they just put it on the auto stainer then sent it >in with all the other slides. > >What did this person learn? NOTHING >And what did ASCP learn? NOTHING (Do they understand they are handing out >HT's to some people who don't know how to do the work?) > >These are just two examples, I could list allot more....But my blood >pressure would go wild. > >Just keep smiling. >Susan > > > > > > > > > > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From wkreider <@t> u.washington.edu Fri Nov 5 13:52:07 2004 From: wkreider <@t> u.washington.edu (Wayne Kreider) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Re: histonet gas bubbles In-Reply-To: References: <418AE91B.7000403@u.washington.edu> Message-ID: <418BD9E7.2020407@u.washington.edu> Phil, Acoustic detection is our primary method. With traditional ultrasound methods at 3-5 MHz, we can detect individual bubbles on the order of 3 microns. Our interest in histology is to develop an independent method to correlate identifiable bubbles and associated biological damage with the acoustic data. We hadn't thought much about acoustic microscopy at higher frequencies because of limitations on penetration depth relative to the size of our treatment area. Also, assessing in detail the state of treated tissue is a relatively new direction for us. However, acoustic microscopy just might provide a nice compromise with information on the bubbles in addition to some of the more traditional histological detail. I'll have to explore this further. We'll see where this leads. We may get some help next week to look at a frozen sample with light microscopy. I think the favored approach by our local help here is to use cryogen cooled isopentane to freeze a relatively large chunk of tissue (order of 2 mm per side). I'm not sure, but they may also require use of a cryoprotectant for their processing. Whatever we see will be questionable, but it will be a new data point for us. As you descibed, there are likely better ways to try to catch and view an intact bubble. This stuff is all related to my dissertation work, so at some point in the next year or two I'll be making a push to obtain this information in some rigorous way. When I do make some significant progress on it, I'll drop a note to keep you up to date. I definitely appreciate your input. Wayne Philip Oshel wrote: > Wayne, > > This triggered another thought: > How big are your bubbles? I forget the details of resolution, etc., > but there is acoustic microscopy. The density differences between > tissue and bubbles would make them good targets for AM. And if the > bubbles are within the resolution (I doubt this, but...), ultrasound. > Depending on the strength and frequency of the sonication used to > produce the bubbles. > Seems like it should be possible to both produce and detect or image > the bubbles by the same general method -- acoustics. > > Phil > >> Linda, >> Thanks for the input. I did get a chance to look at this paper >> briefly. Using these types of markers might be useful if we could >> see them with a high enough resolution. However, given my >> understanding of the method, we would have trouble directly >> differentiating a bubble from dissolved gases, as is our primary >> goal. I'm not just sure where we're headed with this in the future, >> but I'll let you know if we have some success. >> Wayne >> >> lkbauer@unmc.edu wrote: >> >>> >>> >>> Hello Wayne, >>> I recently read an article that may give you some direction. It >>> explains >>> how some gases can be detected by use of trapping agents. This >>> article on >>> nitric oxide( half-life of only a few seconds) summarizes recent >>> methods >>> for visualizing NO in living tissues. Nitric Oxide 9 (2003) 217-228. >>> Nitric >>> oxide imaging in living neuronal tissues using fluorscent probes. >>> Author >>> Oliver von Bohen und Halbach. If you solve this I would be >>> interested in >>> your method. Good luck! >>> >>> >>> Linda(Lin)Bauer >>> Department of Genetics, Cell Biology, and Anatomy >>> 985455 Nebraska Medical Center >>> Omaha, NE 68198-5455 >>> >>> Phone: (402) 559-2863 >>> Fax: (402) 559-4001 >>> Email: lkbauer@unmc.edu >>> >>> >> >> -- >> >> /**********************/ >> >> /Wayne Kreider/ >> >> /University of Washington/ >> >> /Applied Physics Lab / CIMU/ >> >> /106 Old Fisheries/ >> >> /206-543-1324/ >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- /**********************/ /Wayne Kreider/ /University of Washington/ /Applied Physics Lab / CIMU/ /106 Old Fisheries/ /206-543-1324/ From Janet.Bonner <@t> FLHOSP.ORG Fri Nov 5 14:50:17 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Dumbing down the practical???] Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB40DD@fh2k093.fhmis.net> OK- But you guys did not have to WEAR A SKIRT in the dead of winter in New York and walk 1 1/2 miles through 3 feet of snow..Yada, Yada, Yada (Yes- I'm old enough to have been required to wear a skirt) BRRRRRR.... @:) -----Original Message----- From: Ford Royer To: Histo Net Sent: 11/5/2004 2:26 PM Subject: [Histonet] Dumbing down the practical???] ...and I bet it was up-hill BOTH WAYS!!! ;-) ~ Ford ps: You're dating yourself Joe... but then my Medical Technology class was the last class that was required to go through a rotation in Histology, and we had 25 Histo/Cyto questions included on that year's M.T. Registry.... and they were HARD questions! Maybe that's why they dropped Histology from the Med Tech requirements... it was to hard for us . ;-) Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN Poteete, Jacquie A. wrote: >WASTE high snow! What a horrible experience! > >Jacquie Poteete MT(ASCP)QIHC >Lead Technologist, IHC Lab >Saint Francis Hospital >Tulsa, OK > > > >>-----Original Message----- >>From: Joe Nocito [SMTP:JNocito@Pathreflab.com] >>Sent: Friday, November 05, 2004 11:10 AM >>To: TERRI BRAUD; histonet@lists.utsouthwestern.edu >>Subject: RE: [Histonet] Dumbing down the practical??? >> >>I can do one better. >>Two techs came through my lab during a clinical rotation. They were so >>good >>that I hired them. Both took their practical at the same time using the >>same >>tissue from the same autopsy, used the same processor, microtome and >>stainer. I reviewed all their slides as did my medical director. One >>passed, >>one didn't. When I called ASCP to inquire how this could happen, I was >>told >>it would be $100 to have the slides reviewed. I told this person that I >>was >>the supervisor, not the applicant and I wanted to know how this could >>happen. >> I put my name in to become a reviewer, but never received a yes or >>no. >>Do I have some issues with ASCP, you bet. When my techs told me that they >>had to turn in only 9 slides, I was appalled. So I told the same old story >>"When I went to school, I had to walk 5 miles in waste high snow. When I >>did >>my HT, I had to use paraffin molds, process by hand and use a steel >>knife." >>Their response was "was that before cell phones and play stations?" They >>remember seeing a picture of a steel knife in a book once. Geez, am I >>getting old or what? >> >>Joe "The Toe" 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 TERRI >>BRAUD >>Sent: Thursday, November 04, 2004 3:52 PM >>To: histonet@lists.utsouthwestern.edu >>Subject: [Histonet] Dumbing down the practical??? >> >> >>I have a question for all those on the list who have taken the HT >>practical. >> Why do you think that the practical exam does not cover the material >>that >>it used to? Having taken mine in 1981, and then, later as a supervisor, >>having trained 8 OJT's to sucessful completion of their certification, I >>have observed that the practical is not near the test that it used to be. >>There is no CNS tissue, no decalcified tissue, no submission of both a >>special stain and a H&E from the same block. Just one slide, one block, >>one >>stain. Also how is the effect of fully automated special stainers vs >>prepared kits vs "homemade" solutions being taken into account? There >>isn't >>even an informational question pertaining to this so that the ASCP could >>begin to see the impact of the technology in our HT learning process? >>Will >>the technician that can successfully load an autostainer be as effective >>in >>troubleshooting a stain as one who has learned every step by hand? Will it >>even be necessary in the future of Histology? Just curious to hear some >>opinions - Terri >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> >> >> >********* Email Confidentiality Statement ********* >Visit http://www.saintfrancis.com/emailconf.asp > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ 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 JNocito <@t> Pathreflab.com Fri Nov 5 15:00:24 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Re: Dumbing down the practical. In-Reply-To: <004a01c4c36b$6a2a71a0$e0dd3040@Nationwide.net> Message-ID: Susan, every student that rotates through here has molar solutions, normal solutions and per cent solutions questions that they have to pass to receive an acceptable grade for the semester. What's even more fun is I have the recently graduated student who work here teach the new rotating students. That gets my employees to repeat the training they have received. You know, the mother of learning. Joe The Toe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Susan Owens Sent: Friday, November 05, 2004 1:13 PM To: Histonet Subject: [Histonet] Re: Dumbing down the practical. >1. In all those years, there has never been a time that both the H&E and a >special stain has been requested on the same tissue. That would be double >penalizing someone if they had a bad tissue. Well I don't agree. It's been a few years, but I seem to remember that I did special stains and H&E on select blocks when I took the exam(in 1964).. And this thing about a 'double penalizing someone' for a bad tissue...Well they should not of used 'bad tissue' to begin with. And if they didn't know the tissue was 'bad' then they shouldn't of been taking the exam. I believe there are too many new faces in histoland that think they know it all and in truth they don't have a clue. ex: 1. A year or two ago I asked someone to make up a % solution of something(I don't remember what)..I was told they didn't know how. I asked 'how could you not know,you have your HT?' They answered "that they didn't need to know, where they worked before they purchased everything." 2. This year someone I know was taking the practical. According to them they were having a problem getting one of the special stains to work. SO, after several tries, they just put it on the auto stainer then sent it in with all the other slides. What did this person learn? NOTHING And what did ASCP learn? NOTHING (Do they understand they are handing out HT's to some people who don't know how to do the work?) These are just two examples, I could list allot more....But my blood pressure would go wild. Just keep smiling. Susan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jstaruk <@t> masshistology.com Fri Nov 5 16:33:29 2004 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Question for those with private histo labs In-Reply-To: Message-ID: <000001c4c387$79c34c60$6501a8c0@yourw04gtxld67> I am trying to obtain general liability insurance to cover my histology practice and my independent insurance agent claims he contacted 17 different insurers and says he cannot get general liability insurance for my building unless I have professional liability insurance first ($30,000 to $40,000 per year). I told my agent I make slides for third parties (mostly research and academic) and I have no "patients" that can possibly sue me and I don't sign my name on reports. He says I own a medical practice and that magical word "medical" scares off any insurance company. Has anyone else had to deal with something like this? Can some private labs offer me the names of their insurers so I can get some insurance before I have to close my lab? Anyone requiring payment for answering my request need not reply. Thanks Jim ____________________ Jim Staruk Mass Histology Service www.masshistology.com From kspencer <@t> utmem.edu Fri Nov 5 16:53:31 2004 From: kspencer <@t> utmem.edu (Kathleen Spencer) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] curling sections Message-ID: <83AC08C3-2F7D-11D9-BBFF-000393967904@utmem.edu> Hi Everyone, Several months ago I sent out a cry for help because my perfused frozen 20u sections had started curling up in tight little rolls in the buffer, which made it impossible to do IHC on them. We tried EVERYTHING and finally bought a new cryostat, a Leica 1850. It worked well for one month and then the curling started again. The Leica rep thought it had to be the environment and although it sounded crazy, we went with that. So it was the humidity!!!!!!!!!!!! We have moved the cryostat to a small room with a dehumidifier running constantly. The water catcher resevoir was full after 24 hrs! We were amazed and dumbfounded. So Memphis is the humidity capital of the world I guess. Who would have thought? My sections are nice and flat and beautiful now. I want to thank everyone that tried to help and as I promised, am letting you all know that it was the HUMIDITY! I also put a large beaker of EtOH in the cryostat, and by the way, I love the Leica. It is now in a small room in a different building with a dehumidifier running constantly. Cheers, Kathleen Spencer HT (ASCP) Lab Manager/LCM Supervisor UTHSC From lizchlipala <@t> premierhistology.com Fri Nov 5 17:07:52 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Question for those with private histo labs In-Reply-To: <000001c4c387$79c34c60$6501a8c0@yourw04gtxld67> Message-ID: <000201c4c38c$4738d080$76d48a80@AMY> Jim I too own a histology lab. I have liability insurance through The St. Paul. My insurance broker is IMA, which is a large national firm. I did struggle a bit before I obtained insurance. I am required to have liability insurance for the clients in which I do animal work for, and because I lease space at the University of Colorado, but I always thought that liability insurance was important to have. I am a member of the Colorado BioScience Association and through them I was able to get a contact for someone who would provide me insurance. Your state should have some Bioscience association which participates in Bio (Biotechnology Industrial Organization). I found that once I joined that it helped me get in contact with the people I needed to. As far as I know of there are only a few insurance providers such as The St. Paul and Chubb who will insure companies like us. Have your agent call give them a call or try to contact a larger National Broker. Also just to tell you my insurance was not that expensive - around $3500.00 per year, but that can change. A new lab across the hallway from me was quoted $10,000.00 from Chubb. 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 Jim Staruk Sent: Friday, November 05, 2004 3:33 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Question for those with private histo labs I am trying to obtain general liability insurance to cover my histology practice and my independent insurance agent claims he contacted 17 different insurers and says he cannot get general liability insurance for my building unless I have professional liability insurance first ($30,000 to $40,000 per year). I told my agent I make slides for third parties (mostly research and academic) and I have no "patients" that can possibly sue me and I don't sign my name on reports. He says I own a medical practice and that magical word "medical" scares off any insurance company. Has anyone else had to deal with something like this? Can some private labs offer me the names of their insurers so I can get some insurance before I have to close my lab? Anyone requiring payment for answering my request need not reply. Thanks Jim ____________________ Jim Staruk Mass Histology Service www.masshistology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From scoop <@t> mail.nih.gov Fri Nov 5 17:42:53 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] antigen for B-cells in bone marrow Message-ID: Dear Histonetters, Does anyone know of an antibody that works well on B-cells in bone marrow? I'm actually trying to use it on a western blot, but I think many Abs that work on IHC work on western. I've tried CD11a, CD11b, and Pax-5, all of which are supposed to work on western but none did (maybe I need to load more protein). Any suggestions would be greatly appreciated. Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From shahin.zangenehpour <@t> mail.mcgill.ca Sat Nov 6 19:14:26 2004 From: shahin.zangenehpour <@t> mail.mcgill.ca (Shahin Zangenehpour) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Embedding medium for whole monkey brain sections Message-ID: <5DE270EC-305A-11D9-9342-000A95B3CB5E@mail.mcgill.ca> Dear HistoNetters, I am trying to characterise the expression of FMRP (fragile X mental retardation protein) across an entire monkey brain. The goal of the project is to provide a 3D map of FMRP expression. Due to technical constraints with the Ab-Ag interactions, the best way to carry out the histology is to do free-floating IHC on whole-brain monkey sections (50 microns thick). The obvious problem, of course, is that it is impossible to maintain the relative spatial information in the tissue that is essential for constructing a map out of the histological data because whole sections of the brain often come in more than one piece. The only solution that comes to mind is to embed the entire brain in a matrix that will keep the spatial information intact throughout the histological processing and maintains that information for subsequent analysis. However, I have absolutely no clue as to how I can achieve that. Does anyone know of a method that will allow me to embed the entire brain such that I can freeze and section the brain-matrix blocks, carry out the histology using established free-floating IHC techniques, and then mount the brain-matrix sections on glass slides for subsequent analysis? Many thanks, SZ ________________________________________________________________________ ______________ Shahin Zangenehpour, PhD Postdoctoral Fellow | P. 514 398 6151 | F. 514 398 3255 Psychology Dept | McGill University | 1205 Dr Penfield Ave | Montr?al QC Canada H3A 1B1 From lpwenk <@t> sbcglobal.net Sat Nov 6 20:40:12 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] CEU's References: <83AACDB0810528418AA106F9AE9B7F7E50749E@sjhaexc02.sjha.org> Message-ID: <004d01c4c473$1b7d5be0$1bd5d445@domainnotset.invalid> It is only required for those who have taken and passed their HT or HTL exam after Jan. 1, 2004. For those who passed their HT or HTL exam previous to Jan. 1, 2004 - ASCP is NOT requiring the CMP. However, anyone can submit documentation to participate in the CMP program, regardless of when they passed their exam (even an oldie but goodie like me!). Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 P.S. Payment of a fee is also required. ----- Original Message ----- From: "Weems, Joyce" To: ; ; Sent: Friday, November 05, 2004 7:55 AM Subject: RE: [Histonet] CEU's > > 3. ASCP has a new requirement for all people passing a certification exam > after Jan. 1, 2004. Previously, it was - once certified = certified for > life. Now there is a Certification Maintenance Program. (CMP) > > Hi Peggy, > > This was news to me. Does this mean that all of us need to do this or just the ones certified after Jan. 1? > > Thanks, 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 > > > 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 lpwenk <@t> sbcglobal.net Sat Nov 6 21:15:06 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Re: Dumbing down the practical. References: <004a01c4c36b$6a2a71a0$e0dd3040@Nationwide.net> <6.1.2.0.0.20041105134651.028b2780@ander093.email.umn.edu> Message-ID: <001001c4c477$fc738d00$1bd5d445@domainnotset.invalid> For those who remember doing both an H&E and a special stain on the same block - any chance this was BEFORE 1980, when I took my registry exam and started helping students? In my original response to histonet, my note about ASCP not requiring both an H&E and a special stain on the same block was in reference to my time frame (post-1980). I have heard from others, who took their exams in say the 1960's, that did have to do both. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "LuAnn Anderson" To: "Susan Owens" ; "Histonet" Sent: Friday, November 05, 2004 2:52 PM Subject: Re: [Histonet] Re: Dumbing down the practical. > I also remember being required to do specials and H&E's on the same blocks. > If it turned out to be a bad section/sections--or bad tissue--it was back > to the drawing board!! Recuts or new tissue were then in order!! AND, not > to date myself, but everything was done by hand with the exception of > processing!! I too,believe that it is important have that "hands on" know > how for troubleshooting purposes. JMHO. > LuAnn > > > > At 01:12 PM 11/5/04, Susan Owens wrote: > > >1. In all those years, there has never been a time that both the H&E and a > > >special stain has been requested on the same tissue. That would be double > > >penalizing someone if they had a bad tissue. > > > > > > > >Well I don't agree. It's been a few years, but I seem to remember that I did > >special stains and H&E on select blocks when I took the exam(in 1964).. And > >this thing about a 'double penalizing someone' for a bad tissue...Well they > >should not of used 'bad tissue' to begin with. And if they didn't know the > >tissue was 'bad' then they shouldn't of been taking the exam. > >I believe there are too many new faces in histoland that think they know it > >all and in truth they don't have a clue. > > > >ex: 1. A year or two ago I asked someone to make up a % solution of > >something(I don't remember what)..I was told they didn't know how. I asked > >'how could you not know,you have your HT?' They answered "that they didn't > >need to know, where they worked before they purchased everything." > > 2. This year someone I know was taking the practical. According to > >them they were having a problem getting one of the special stains to work. > >SO, after several tries, they just put it on the auto stainer then sent it > >in with all the other slides. > > > >What did this person learn? NOTHING > >And what did ASCP learn? NOTHING (Do they understand they are handing out > >HT's to some people who don't know how to do the work?) > > > >These are just two examples, I could list allot more....But my blood > >pressure would go wild. > > > >Just keep smiling. > >Susan > > > > > > > > > > > > > > > > > > > > > > > > > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@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 brian.dias <@t> mail.utexas.edu Sat Nov 6 21:13:22 2004 From: brian.dias <@t> mail.utexas.edu (Brian Dias) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Sections falling off slides Message-ID: <5.2.0.9.2.20041106210425.00bccae8@mail.utexas.edu> hi i'm trying to optimize my immunohistochemistry protocol on slides and am runnning into a few problems. 1. i'd like to know if anyone has any strong opinions (either positive/negative) about conducting such protocols on slides v/s free-floating. i'd like to be able to do this on slides because mounting my sections after performing a free-floating technique is virutally impossible due to the fragile nature and small size of of my sections. 2. i run into the problem of my sectiions falling off the slide whilst doing my ICC the brain is dissected form the skull post-perfusion with saline and 4% paraformaldehyde it is then stored in 4% paraformaldehyde for 3 days at 4 C then transferred to 20%sucrose/1X PBS and stored at 4 C for 3 days before sectioning on the cryostat at a 30-50um thickness and mounted on Fisher Probe-On Superfrost Plus slides. slides are stored at -80C after they've dried (approx. 20 mins). just before (the night before/2 hours before) performing the ICC, i remove the slides from the freezer and allow to air-dry. immerse slides in 4% paraformaldehyde for 10 mins at RT in a dish 1X PBS wash at RT in a dish then normal ICC steps by overlaying solution on slides and here's where sections come peeling off. i'd appreciate any input on the matter thanks a ton regards brian ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ Brian George Dias Graduate Student (Crews lab) Office Phone #: 512-475-6738 The University of Texas at Austin E-mail: brian.dias@mail.utexas.edu Institute for Neuroscience Website: https://webspace.utexas.edu/bgd85/b.htm Patterson 56 1 University Station Stop C0930 Austin, TX 78712 ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ From lpwenk <@t> sbcglobal.net Sat Nov 6 21:44:12 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] CEU's References: Message-ID: <002501c4c47c$0f0fd0a0$1bd5d445@domainnotset.invalid> I don't see why not. Obtaining a QIHC is 12 points. (Of course, you already HAVE the QIHC, BEFORE signing up for the CMP. So I don't think it would count for you. Only if you obtained advanced certification AFTER signing up for the CMP. I'm guessing.) But remember, I'm not the authority. ASCP Board of Registry is. If anyone is curious or wants information specific to their situation, call 312-738-1336, press "O" for operator, and ask for the BOR. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Joe Nocito" To: "Bonnie Whitaker" ; Sent: Friday, November 05, 2004 12:11 PM Subject: RE: [Histonet] CEU's > Bonnie, > I'll do it if the same CEUs I take for my QIHC can be used for the CMP > > 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 Bonnie > Whitaker > Sent: Friday, November 05, 2004 10:23 AM > To: histonet@pathology.swmed.edu > Subject: RE: [Histonet] CEU's > > > How many folks out there plan on doing the optional (for those already > registered) Certification Maintenance Program? I think it's a good idea, > and I plan on doing it, but I just wanted to know what the general feeling > is. Will anyone require that of their employees? > > Bonnie Whitaker > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, > Janet > Sent: Friday, November 05, 2004 9:54 AM > To: 'Jennifer MacDonald '; 'lpwenk@sbcglobal.net ' > Cc: 'KMB1904@aol.com '; 'histonet@pathology.swmed.edu ' > Subject: RE: [Histonet] CEU's > > > Well, Florida sure does - I order mine through Anderson Continuing > Education ; 3268 Ramos Circle / P.O.Box 276297 ; Sacramento, Ca 95827 > Phone: 1-800-532-2332 / FAX 1-916-856-5066 > They're really easy to use. You buy the book and they send a test booklet > of multiple choice questions and a scantron sheet - which you can fax when > you're done. You'll get a certificate in the mail within a week. Each > course is mostly 24 CEUs with some exceptions that are 12 or 36. There's > also one out of Miami, FL but I don't have the info on that. > Give them a call!! Janet > > > -----Original Message----- > From: Jennifer MacDonald > To: lpwenk@sbcglobal.net > Cc: KMB1904@aol.com; histonet@pathology.swmed.edu > Sent: 11/5/2004 1:46 AM > Subject: Re: [Histonet] CEU's > > > California has no requirements for Histotechnicians for certifica > tion or continuing education. > > > > Jennifer MacDonald > -----histonet-bounces@lists.utsouthwestern.= edu wrote: ----- > > To: , From: > > Sent by: histonet-bounces@lists.u= tsouthwestern.edu > Date: 11/04/2004 06:15PM > Subject: Re: [Histonet] CE= U's > Multiple faceted question. I'll = try to answer it as best as I > can. > 1.There is NO national requiremen= t to have CEU to continue to > work in > histology (or cytology or med tech,= etc.). Most states have NO > requirements > (Florida and maybe Califo= rnia are exceptions). > 2. Hospitals or your place of employment or ev= en you lab itself > can set any > requirement they want, including no requir= ement. > 3. ASCP has a new requirement for all people passing a certif > ication exam > after Jan. 1, 2004. Previously, it was - once certified = > certified for > life. Now there is a Certification Maintenance Program. (= CMP) > [1]= http://www.ascp.org/bor/cmp/index > Now, if you want to maintain y= our certification status through > ASCP, you > must have 36 points of contin= uing education every 3 year, submit > a form with > this information along w= ith a renewal fee (currently $50). > Documentation > such as CEU forms are n= ot required, unless you are one of the > randomly > picked to be audited. Of= the 36 points, for HT/HTL, 1 point must > be on > safety, 2 points in your = area of specialty (i.e., > histotechnology) and the > rest of the points can= be in your area of specialty, management, > education, > or any related lab= area of interest. > Points can be awarded for many types of CE, and e= ach type can be > worth > different number of points. > For example: > - 1= hour workshop =3D 1 point > - 1 hour teleconference = 1 point > - 1 se= mester hour of college =3D 15 points > - competency assessment by your emp= loyer =3D 2 points per year > - authoring an article in a peer reviewed jo= urnal (such as NSH > Journal of > Histotechnology or ASCP Laboratory Medicin= e) =3D 5 points. > 4. Ways to get CE: > Check in the ASCP CMP pac= ket (see above link, click on right > side). Page 6 > has lots of suggestion= s. > - attending state meetings > - attending national meetings > - = in-services held at your institution > - teleconferences (available throug= h NSH (301-262-6221) and ASCP > (312-738-1336) > - college classes (relat= ed to labs) > - safety classes at your institution > - competency assessm= ent > - present at state and national meetings > - write articles for his= totechnology journals > - attend vendor workshops > 36 points in 3 ye= ars really isn't a lot to have to get. And you > don't have > to spend a lot= of money. In-services are free - just make certain > you have > documentati= on (i.e. sign in sheet). Most places have you do > safety quizzes - > fire, = electrical, blood borne pathogens, etc. They would count. > Peggy A. W= enk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > ----- Original Message ----- > From: > To: > Sent: Thursday, November 04, 2004 5:39 PM > Subjec= t: [Histonet] CEU's > > Does anyone know where you can ge= t CEU credits for histology? > Are we > > obligated to have so man= y a year? And if so, how many?? I > have never > seen this > &g= t; information written anywhere. > > > > > > ______ ______________________ 5F__= > =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F > > Histonet mailing l= ist > > Histonet@lists.utsouthwestern.edu > > [2]http://li= sts.utsouthwestern.edu/mailman/listinfo/histonet > ___ ______________________ 5F__= > =5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F=5F > Histonet maili= ng list > Histonet@lists.utsouthwestern.edu > [3]http://lists.ut= southwestern.edu/mailman/listinfo/histonet > > <= /FONT> > References > > 1. 3D"http://www.ascp.org/bor/cmp/index" > 2. 3D"http://lis=/ > 3. 3D"http://lists.uts=/ _______________________________________________ > 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 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mprice26 <@t> juno.com Sun Nov 7 06:20:27 2004 From: mprice26 <@t> juno.com (marsha r price) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] RE:Resubscribe?? Message-ID: <20041107.062028.4980.0.mprice26@juno.com> I would like to resubscribe to histonet. I sent a job announcement out over the histonet. Someone e-mailed me back and said that my e-mail message contained several viruses. I sent the message from computer at work. I reported problem to office manager and they apparently fixed the problem. I have virus protection software at my home computer. Please tell me how I can start receiving messages from Histonet again. Marsha Price ________________________________________________________________ Juno Platinum $9.95. Juno SpeedBand $14.95. Sign up for Juno Today at http://www.juno.com! Look for special offers at Best Buy stores. From jo-ann-e.bader <@t> staff.mcgill.ca Mon Nov 8 08:18:35 2004 From: jo-ann-e.bader <@t> staff.mcgill.ca (jo-ann-e.bader@staff.mcgill.ca) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Xylene substitute problems Message-ID: <1099923515.418f803b0e85a@webmail.mcgill.ca> Good morning to all histonetters, At around May of this year I had to change from xylene to a xylene substitute and have been hhaving problems eversince. I am processing all kinds of mouse tissue and I find that the tissue seems to be softer, produces more wrinkles and is generally more difficult to cut. Currently I am using 3 baths at 1 hour each. I have been using diffrent times throughout the summer. If anyone has any pointers I would appreciate it. Thanks Jo-Ann From JNocito <@t> Pathreflab.com Mon Nov 8 08:57:07 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Dumbing down the practical???] In-Reply-To: <07AB60D5D7B9754EBF56F360F98D083DEB40DD@fh2k093.fhmis.net> Message-ID: Janet, was that in up state New York or on the Island? Joe the Toe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bonner, Janet Sent: Friday, November 05, 2004 2:50 PM To: 'Ford Royer '; 'Histo Net ' Subject: RE: [Histonet] Dumbing down the practical???] OK- But you guys did not have to WEAR A SKIRT in the dead of winter in New York and walk 1 1/2 miles through 3 feet of snow..Yada, Yada, Yada (Yes- I'm old enough to have been required to wear a skirt) BRRRRRR.... @:) -----Original Message----- From: Ford Royer To: Histo Net Sent: 11/5/2004 2:26 PM Subject: [Histonet] Dumbing down the practical???] ...and I bet it was up-hill BOTH WAYS!!! ;-) ~ Ford ps: You're dating yourself Joe... but then my Medical Technology class was the last class that was required to go through a rotation in Histology, and we had 25 Histo/Cyto questions included on that year's M.T. Registry.... and they were HARD questions! Maybe that's why they dropped Histology from the Med Tech requirements... it was to hard for us . ;-) Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN Poteete, Jacquie A. wrote: >WASTE high snow! What a horrible experience! > >Jacquie Poteete MT(ASCP)QIHC >Lead Technologist, IHC Lab >Saint Francis Hospital >Tulsa, OK > > > >>-----Original Message----- >>From: Joe Nocito [SMTP:JNocito@Pathreflab.com] >>Sent: Friday, November 05, 2004 11:10 AM >>To: TERRI BRAUD; histonet@lists.utsouthwestern.edu >>Subject: RE: [Histonet] Dumbing down the practical??? >> >>I can do one better. >>Two techs came through my lab during a clinical rotation. They were so >>good >>that I hired them. Both took their practical at the same time using the >>same >>tissue from the same autopsy, used the same processor, microtome and >>stainer. I reviewed all their slides as did my medical director. One >>passed, >>one didn't. When I called ASCP to inquire how this could happen, I was >>told >>it would be $100 to have the slides reviewed. I told this person that I >>was >>the supervisor, not the applicant and I wanted to know how this could >>happen. >> I put my name in to become a reviewer, but never received a yes or >>no. >>Do I have some issues with ASCP, you bet. When my techs told me that they >>had to turn in only 9 slides, I was appalled. So I told the same old story >>"When I went to school, I had to walk 5 miles in waste high snow. When I >>did >>my HT, I had to use paraffin molds, process by hand and use a steel >>knife." >>Their response was "was that before cell phones and play stations?" They >>remember seeing a picture of a steel knife in a book once. Geez, am I >>getting old or what? >> >>Joe "The Toe" 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 TERRI >>BRAUD >>Sent: Thursday, November 04, 2004 3:52 PM >>To: histonet@lists.utsouthwestern.edu >>Subject: [Histonet] Dumbing down the practical??? >> >> >>I have a question for all those on the list who have taken the HT >>practical. >> Why do you think that the practical exam does not cover the material >>that >>it used to? Having taken mine in 1981, and then, later as a supervisor, >>having trained 8 OJT's to sucessful completion of their certification, I >>have observed that the practical is not near the test that it used to be. >>There is no CNS tissue, no decalcified tissue, no submission of both a >>special stain and a H&E from the same block. Just one slide, one block, >>one >>stain. Also how is the effect of fully automated special stainers vs >>prepared kits vs "homemade" solutions being taken into account? There >>isn't >>even an informational question pertaining to this so that the ASCP could >>begin to see the impact of the technology in our HT learning process? >>Will >>the technician that can successfully load an autostainer be as effective >>in >>troubleshooting a stain as one who has learned every step by hand? Will it >>even be necessary in the future of Histology? Just curious to hear some >>opinions - Terri >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> >> >> >********* Email Confidentiality Statement ********* >Visit http://www.saintfrancis.com/emailconf.asp > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ 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 gentras <@t> vetmed.auburn.edu Mon Nov 8 09:02:44 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:16 2005 Subject: Fwd: { SPAM 2 }::[Histonet] Xylene substitute problems Message-ID: <6.0.1.1.0.20041108085611.01b018b8@mailhost.vetmed.auburn.edu> Good morning! Please, I'm wondering which substitute are you using and what type tissues (CNS/extraneural or both) are you processing? For the past 20+ years I have been using Fisher Scientific's Tissue Clear III and have not experienced any problems. Although, the majority of my tissues are CNS, all or my processing is done using it: even the mouse brains & extraneurals. Best wishes. Atoska >From: jo-ann-e.bader@staff.mcgill.ca >To: Histonet >Subject: { SPAM 2 }::[Histonet] Xylene substitute problems > >Good morning to all histonetters, > >At around May of this year I had to change from xylene to a xylene substitute >and have been hhaving problems eversince. I am processing all kinds of mouse >tissue and I find that the tissue seems to be softer, produces more wrinkles >and is generally more difficult to cut. Currently I am using 3 baths at 1 >hour >each. I have been using diffrent times throughout the summer. If anyone has >any pointers I would appreciate it. > >Thanks > >Jo-Ann From Terry.Coaker <@t> nuth.northy.nhs.uk Mon Nov 8 09:30:11 2004 From: Terry.Coaker <@t> nuth.northy.nhs.uk (Coaker, Terry) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Histological dissection of bone Message-ID: For many years we have "trimmed" bone samples on a band-saw to obtain slices for decalcification. The equipment needs replacing. A risk assessment indicated that putting a long bone through a band-saw in this way could lead to someone getting hurt! Is there a safer way? Diamond saws are fine for trimming small pieces but what about femur and humerus? How do people handle them safely? Thankyou Terry Terry Coaker Histopathology Operations Manager Cellular Pathology Royal Victoria Infirmary Queen Victoria Road Newcastle upon Tyne NE1 4LP 0191 282 9120 From MDiCarlo <@t> KaleidaHealth.Org Mon Nov 8 10:46:26 2004 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Histological dissection of bone Message-ID: Terry, I also use a band saw for cutting bones of all different sizes, such as femur, tibia, etc. People have raved about the Mar-Med Band Saw that costs $810 but I don't think it is large enough for large sections of bone such as a whole tibia or fibula that we have to cut up. It is an 18-pound tabletop band saw with a diamond blade and is about the size of a microscope. I will be interested to hear what everyone has to say. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: Coaker, Terry [mailto:Terry.Coaker@nuth.northy.nhs.uk] Sent: Monday, November 08, 2004 10:30 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Histological dissection of bone For many years we have "trimmed" bone samples on a band-saw to obtain slices for decalcification. The equipment needs replacing. A risk assessment indicated that putting a long bone through a band-saw in this way could lead to someone getting hurt! Is there a safer way? Diamond saws are fine for trimming small pieces but what about femur and humerus? How do people handle them safely? Thankyou Terry Terry Coaker Histopathology Operations Manager Cellular Pathology Royal Victoria Infirmary Queen Victoria Road Newcastle upon Tyne NE1 4LP 0191 282 9120 _______________________________________________ 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 kosmicdog <@t> hotmail.com Mon Nov 8 10:47:18 2004 From: kosmicdog <@t> hotmail.com (jason m) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Immuno Artifact? Message-ID: Can anyone identify the cause of this black circle artifact? It is small black circles that look sort of like air bubbles only much too small which have been turning up on my immuno's this week. I have changed all solutions but still have the problem. Our lab uses toluene instead of xylenes, I am wondering if that is the problem. Also I use Tween-20 in my citrate buffer and I wonder if that could be the problem. At any rate I will be checking both of these but if anyone can help It will be appreciated immensely. I have sent a photograph of the artifact to the the list serv image archive (see link below) under the filename "artifact.jpg". http://www.histonet.org/site_images_frame.asp Thanks again, Jason From vbaker60 <@t> yahoo.com Mon Nov 8 11:12:08 2004 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Looking for Tissue Tek manual staining set up Message-ID: <20041108171208.74667.qmail@web50110.mail.yahoo.com> Hi! Thanks to everyone who responded to my Belair request, it was a big help. I was wondering if anyone had a vendor that they could recommend for this staining set up. So far I've only been able to identify 1 vendor and I need at least one more quote. Thanks in advance. Vikki Baker Mt. Sinai School of Medicine NY, NY 10029 __________________________________ Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com From Terry.Marshall <@t> rothgen.nhs.uk Mon Nov 8 11:12:10 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Immuno Artifact? Message-ID: That surely is entrapped air under the coverslip. 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: jason m [mailto:kosmicdog@hotmail.com] Sent: 08 November 2004 16:47 To: histonet@pathology.swmed.edu Subject: [Histonet] Immuno Artifact? Can anyone identify the cause of this black circle artifact? It is small black circles that look sort of like air bubbles only much too small which have been turning up on my immuno's this week. I have changed all solutions but still have the problem. Our lab uses toluene instead of xylenes, I am wondering if that is the problem. Also I use Tween-20 in my citrate buffer and I wonder if that could be the problem. At any rate I will be checking both of these but if anyone can help It will be appreciated immensely. I have sent a photograph of the artifact to the the list serv image archive (see link below) under the filename "artifact.jpg". http://www.histonet.org/site_images_frame.asp Thanks again, Jason _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bhewlett <@t> cogeco.ca Mon Nov 8 11:36:53 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Immuno Artifact? References: Message-ID: <001f01c4c5b9$8a1ec330$6400a8c0@mainbox> I agree, but it could also be water since the Becke lines do not appear quite wide enough for air. Bryan Hewlett, ART, MLT Consultant Technologist Quality Management Program-Laboratory Services Toronto, Ontario, Canada. ----- Original Message ----- From: "Marshall Terry Dr, Consultant Histopathologist" To: "jason m" ; Sent: Monday, November 08, 2004 12:12 PM Subject: RE: [Histonet] Immuno Artifact? That surely is entrapped air under the coverslip. 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: jason m [mailto:kosmicdog@hotmail.com] Sent: 08 November 2004 16:47 To: histonet@pathology.swmed.edu Subject: [Histonet] Immuno Artifact? Can anyone identify the cause of this black circle artifact? It is small black circles that look sort of like air bubbles only much too small which have been turning up on my immuno's this week. I have changed all solutions but still have the problem. Our lab uses toluene instead of xylenes, I am wondering if that is the problem. Also I use Tween-20 in my citrate buffer and I wonder if that could be the problem. At any rate I will be checking both of these but if anyone can help It will be appreciated immensely. I have sent a photograph of the artifact to the the list serv image archive (see link below) under the filename "artifact.jpg". http://www.histonet.org/site_images_frame.asp Thanks again, Jason _______________________________________________ Histonet mailing list Histonet@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 mrsgbd2001 <@t> yahoo.com Mon Nov 8 12:21:33 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Immuno Artifact? In-Reply-To: Message-ID: <20041108182134.84211.qmail@web52701.mail.yahoo.com> Looks to me like it wasn't probably dehydrated, and those are water bubbles. Gareth jason m wrote: Can anyone identify the cause of this black circle artifact? It is small black circles that look sort of like air bubbles only much too small which have been turning up on my immuno's this week. I have changed all solutions but still have the problem. Our lab uses toluene instead of xylenes, I am wondering if that is the problem. Also I use Tween-20 in my citrate buffer and I wonder if that could be the problem. At any rate I will be checking both of these but if anyone can help It will be appreciated immensely. I have sent a photograph of the artifact to the the list serv image archive (see link below) under the filename "artifact.jpg". http://www.histonet.org/site_images_frame.asp Thanks again, Jason _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com From ander093 <@t> tc.umn.edu Mon Nov 8 12:49:50 2004 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Immuno Artifact? In-Reply-To: <001f01c4c5b9$8a1ec330$6400a8c0@mainbox> References: <001f01c4c5b9$8a1ec330$6400a8c0@mainbox> Message-ID: <6.1.2.0.0.20041108124416.028bb730@ander093.email.umn.edu> My guess is water in the toluene/xylene from the dehydration and clearing run down. He can test this by changing the alcohols and xylenes--removing the coverslips--taking slides back through xylenes, and alcohols to 95% and then re-running them through fresh alcohols and xylenes/toluenes and coverslipping. If it is water, it will disappear. LuAnn At 11:36 AM 11/8/04, Bryan Hewlett wrote: >I agree, but it could also be water since the Becke lines do not appear >quite wide enough for air. > >Bryan Hewlett, ART, MLT >Consultant Technologist >Quality Management Program-Laboratory Services >Toronto, Ontario, >Canada. > >----- Original Message ----- >From: "Marshall Terry Dr, Consultant Histopathologist" > >To: "jason m" ; >Sent: Monday, November 08, 2004 12:12 PM >Subject: RE: [Histonet] Immuno Artifact? > > >That surely is entrapped air under the coverslip. > >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: jason m [mailto:kosmicdog@hotmail.com] >Sent: 08 November 2004 16:47 >To: histonet@pathology.swmed.edu >Subject: [Histonet] Immuno Artifact? > > >Can anyone identify the cause of this black circle artifact? It is small >black circles that look sort of like air bubbles only much too small which >have been turning up on my immuno's this week. I have changed all solutions >but still have the problem. Our lab uses toluene instead of xylenes, I am >wondering if that is the problem. Also I use Tween-20 in my citrate buffer >and I wonder if that could be the problem. At any rate I will be checking >both of these but if anyone can help It will be appreciated immensely. I >have sent a photograph of the artifact to the the list serv image archive >(see link below) under the filename "artifact.jpg". > >http://www.histonet.org/site_images_frame.asp > >Thanks again, >Jason > > > >_______________________________________________ >Histonet mailing list >Histonet@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 Patty.Lott <@t> ORTHO.UAB.EDU Mon Nov 8 13:04:12 2004 From: Patty.Lott <@t> ORTHO.UAB.EDU (Patty Lott) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Shandon Autosharp Message-ID: <85F6C7A1330E794DB8540AFD001CC77E05328BA0@rosco.ortho.uab.edu> Is anyone still using a Shandon Autosharp for sharpening tungsten carbide knives? I need a copy of the handbook, and I need to see what reagents (pastes, etc.) I would need to buy to put an old one which we found into service again! Patty Lott, Laboratory Supervisor Orthopaedic Research Laboratory Center for Metabolic Bone Disease Laboratory University of Alabama at Birmingham LHRB B37 0007 1919 7th Ave. South Birmingham, AL 35294 (205) 934-2007 From JCarpenter764 <@t> aol.com Mon Nov 8 13:24:17 2004 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Thank you!!! Message-ID: <156.437c0f00.2ec121e1@aol.com> I just wanted to thank everyone for their help with all my questions. I took my written exam nov. 1 and passed!! I couldn't believe it...thanks again Jennell From darrenj <@t> medica.co.nz Mon Nov 8 13:46:57 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Shandon Autosharp In-Reply-To: <85F6C7A1330E794DB8540AFD001CC77E05328BA0@rosco.ortho.uab.edu> Message-ID: <000601c4c5cb$b52e4d40$c864a8c0@medica.co.nz> Hi Patty, I suggest you email Thermo Electron (formerly Thermo Shandon) at clinical.diagnostics@thermo.com for a catalogue.Also check their website thermo.com/shandon. The Shandon Autosharp is still available as are all the associated consumables. Thanks Darren James Technical Representative Medica Pacifica Ltd Office Address: 1A 153 Stoddard Road, Mt Roskill, Auckland Postal Address: PO Box 24-421, Royal Oak, Auckland 1030 Ph: +64 9 629 0823 Fax: +64 9 629 0542 Mob: + 64 21 633 820 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patty Lott Sent: Tuesday, 9 November 2004 8:04 a.m. To: histonet@pathology.swmed.edu Subject: [Histonet] Shandon Autosharp Is anyone still using a Shandon Autosharp for sharpening tungsten carbide knives? I need a copy of the handbook, and I need to see what reagents (pastes, etc.) I would need to buy to put an old one which we found into service again! Patty Lott, Laboratory Supervisor Orthopaedic Research Laboratory Center for Metabolic Bone Disease Laboratory University of Alabama at Birmingham LHRB B37 0007 1919 7th Ave. South Birmingham, AL 35294 (205) 934-2007 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jengirl1014 <@t> yahoo.com Mon Nov 8 14:13:38 2004 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] slide stainers and lablers Message-ID: <20041108201338.83244.qmail@web60610.mail.yahoo.com> I'm looking for any info on slide stainers and slide lablers. Our lab is looking to purchase one of each. All responses will be greatly appreciated. Thanks to everyone for helping me out whether it is now or in the past. You guys ar egreat and I love this forum! --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com From MDiCarlo <@t> KaleidaHealth.Org Mon Nov 8 13:42:20 2004 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Shandon Autosharp Message-ID: Patty, I use the Shandon Autosharp V for sharpening C profile blades, haven't used it for tungsten yet. I am sure you can call Thermo-Shandon and ask them to fax you a manual (Operator Guide) for sharpening. Thermo-Shandon's number is 1-800-547-7429. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 -----Original Message----- From: Patty Lott [mailto:Patty.Lott@ORTHO.UAB.EDU] Sent: Monday, November 08, 2004 14:04 To: histonet@pathology.swmed.edu Subject: [Histonet] Shandon Autosharp Is anyone still using a Shandon Autosharp for sharpening tungsten carbide knives? I need a copy of the handbook, and I need to see what reagents (pastes, etc.) I would need to buy to put an old one which we found into service again! Patty Lott, Laboratory Supervisor Orthopaedic Research Laboratory Center for Metabolic Bone Disease Laboratory University of Alabama at Birmingham LHRB B37 0007 1919 7th Ave. South Birmingham, AL 35294 (205) 934-2007 _______________________________________________ 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 lange <@t> kennedykrieger.org Mon Nov 8 14:48:43 2004 From: lange <@t> kennedykrieger.org (Mollie Lange) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] block storage and eradastain Message-ID: Hello Everyone, I have two questions. First, I need to store approximately 2000 (yes, 2000!) paraffin blocks, each measuring approx 24X40X20mm (not cassette size). I am looking for sturdy cardboard boxes a little smaller than shoe box size. Readily available freezer boxes are too small, and other conventional boxes seem too flimsy. Does anyone have a good source for boxes? Maybe a vendor who makes boxes to spec? Second- are Eradastain and Eradosol still available? We used to use this in the Dark Ages to remove stain residue from glassware and fingers. Is there a good substitute, homemade or commercially available? I am in the research side of neurohistology and neuropathology and have gotten tons of useful info from the Histonet. Thanks for any suggestions, Mollie Lange Neuroscience Research Laboratory Kennedy Krieger Institute Baltimore, MD From bills <@t> icpmr.wsahs.nsw.gov.au Mon Nov 8 14:49:17 2004 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Histological dissection of bone In-Reply-To: Message-ID: <000001c4c5d4$6a5301e0$83a7080a@wsahs.nsw.gov.au> Terry, For the last 15 years we have been using a "gem" saw just like they use to cut rocks etc prior to polishing. It has a diamond impregated thin blade 1mm thick and 30cm diameter and a clamping mechanism to hold the material (needs some modification for the shape of bone). The specimen is automatically driven towards the blade. No need for fingers anywhere near it. It will cut specimens up to 12cm in height. We replace the blade about every two years. It gives a beautiful clean cut with little heat being generated during the cutting. Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Coaker, Terry Sent: Tuesday, 09 November 2004 2:30 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Histological dissection of bone For many years we have "trimmed" bone samples on a band-saw to obtain slices for decalcification. The equipment needs replacing. A risk assessment indicated that putting a long bone through a band-saw in this way could lead to someone getting hurt! Is there a safer way? Diamond saws are fine for trimming small pieces but what about femur and humerus? How do people handle them safely? Thankyou Terry Terry Coaker Histopathology Operations Manager Cellular Pathology Royal Victoria Infirmary Queen Victoria Road Newcastle upon Tyne NE1 4LP 0191 282 9120 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From liz <@t> premierlab.com Mon Nov 8 15:06:24 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] block storage and eradastain In-Reply-To: Message-ID: <000c01c4c5d6$ce500380$76d48a80@AMY> Mollie Try Reliable Office Products, that's where I buy my boxes, they have a bunch of different sizes. Look under corrugated mailers or shipping boxes. 1-800-735-5000 or www.reliable.com 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 Mollie Lange Sent: Monday, November 08, 2004 1:49 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] block storage and eradastain Hello Everyone, I have two questions. First, I need to store approximately 2000 (yes, 2000!) paraffin blocks, each measuring approx 24X40X20mm (not cassette size). I am looking for sturdy cardboard boxes a little smaller than shoe box size. Readily available freezer boxes are too small, and other conventional boxes seem too flimsy. Does anyone have a good source for boxes? Maybe a vendor who makes boxes to spec? Second- are Eradastain and Eradosol still available? We used to use this in the Dark Ages to remove stain residue from glassware and fingers. Is there a good substitute, homemade or commercially available? I am in the research side of neurohistology and neuropathology and have gotten tons of useful info from the Histonet. Thanks for any suggestions, Mollie Lange Neuroscience Research Laboratory Kennedy Krieger Institute Baltimore, MD _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From weneng2004 <@t> yahoo.com Mon Nov 8 16:08:29 2004 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] PCNA Ab Message-ID: <20041108220829.83994.qmail@web53403.mail.yahoo.com> Hello, I was asked to do PCNA IHC on formalin fixed paraffin embedded mice skin sections. Does anyone know which antibody works great? Thanks in advance, Wen __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From weneng2004 <@t> yahoo.com Mon Nov 8 16:08:42 2004 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] PCNA Ab Message-ID: <20041108220842.11519.qmail@web53401.mail.yahoo.com> Hello, I was asked to do PCNA IHC on formalin fixed paraffin embedded mice skin sections. Does anyone know which antibody works great? Thanks in advance, Wen --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com From Janet.Bonner <@t> FLHOSP.ORG Mon Nov 8 16:45:21 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Dumbing down the practical???] Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB40E0@fh2k093.fhmis.net> This was in Rochester where the Canadian winds blow off of Lake Ontario - AND - that's why I'm in Florida now.(Thinking N.Carolina might not be bad since this past summer. @:) -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Monday, November 08, 2004 9:57 AM To: Bonner, Janet; 'Ford Royer '; 'Histo Net ' Subject: RE: [Histonet] Dumbing down the practical???] Janet, was that in up state New York or on the Island? Joe the Toe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bonner, Janet Sent: Friday, November 05, 2004 2:50 PM To: 'Ford Royer '; 'Histo Net ' Subject: RE: [Histonet] Dumbing down the practical???] OK- But you guys did not have to WEAR A SKIRT in the dead of winter in New York and walk 1 1/2 miles through 3 feet of snow..Yada, Yada, Yada (Yes- I'm old enough to have been required to wear a skirt) BRRRRRR.... @:) -----Original Message----- From: Ford Royer To: Histo Net Sent: 11/5/2004 2:26 PM Subject: [Histonet] Dumbing down the practical???] ...and I bet it was up-hill BOTH WAYS!!! ;-) ~ Ford ps: You're dating yourself Joe... but then my Medical Technology class was the last class that was required to go through a rotation in Histology, and we had 25 Histo/Cyto questions included on that year's M.T. Registry.... and they were HARD questions! Maybe that's why they dropped Histology from the Med Tech requirements... it was to hard for us . ;-) Ford M. Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN Poteete, Jacquie A. wrote: >WASTE high snow! What a horrible experience! > >Jacquie Poteete MT(ASCP)QIHC >Lead Technologist, IHC Lab >Saint Francis Hospital >Tulsa, OK > > > >>-----Original Message----- >>From: Joe Nocito [SMTP:JNocito@Pathreflab.com] >>Sent: Friday, November 05, 2004 11:10 AM >>To: TERRI BRAUD; histonet@lists.utsouthwestern.edu >>Subject: RE: [Histonet] Dumbing down the practical??? >> >>I can do one better. >>Two techs came through my lab during a clinical rotation. They were so >>good >>that I hired them. Both took their practical at the same time using the >>same >>tissue from the same autopsy, used the same processor, microtome and >>stainer. I reviewed all their slides as did my medical director. One >>passed, >>one didn't. When I called ASCP to inquire how this could happen, I was >>told >>it would be $100 to have the slides reviewed. I told this person that I >>was >>the supervisor, not the applicant and I wanted to know how this could >>happen. >> I put my name in to become a reviewer, but never received a yes or >>no. >>Do I have some issues with ASCP, you bet. When my techs told me that they >>had to turn in only 9 slides, I was appalled. So I told the same old story >>"When I went to school, I had to walk 5 miles in waste high snow. When I >>did >>my HT, I had to use paraffin molds, process by hand and use a steel >>knife." >>Their response was "was that before cell phones and play stations?" They >>remember seeing a picture of a steel knife in a book once. Geez, am I >>getting old or what? >> >>Joe "The Toe" 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 TERRI >>BRAUD >>Sent: Thursday, November 04, 2004 3:52 PM >>To: histonet@lists.utsouthwestern.edu >>Subject: [Histonet] Dumbing down the practical??? >> >> >>I have a question for all those on the list who have taken the HT >>practical. >> Why do you think that the practical exam does not cover the material >>that >>it used to? Having taken mine in 1981, and then, later as a supervisor, >>having trained 8 OJT's to sucessful completion of their certification, I >>have observed that the practical is not near the test that it used to be. >>There is no CNS tissue, no decalcified tissue, no submission of both a >>special stain and a H&E from the same block. Just one slide, one block, >>one >>stain. Also how is the effect of fully automated special stainers vs >>prepared kits vs "homemade" solutions being taken into account? There >>isn't >>even an informational question pertaining to this so that the ASCP could >>begin to see the impact of the technology in our HT learning process? >>Will >>the technician that can successfully load an autostainer be as effective >>in >>troubleshooting a stain as one who has learned every step by hand? Will it >>even be necessary in the future of Histology? Just curious to hear some >>opinions - Terri >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> >> >> >********* Email Confidentiality Statement ********* >Visit http://www.saintfrancis.com/emailconf.asp > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ 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 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 mprice26 <@t> juno.com Mon Nov 8 17:41:37 2004 From: mprice26 <@t> juno.com (marsha r price) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] RE: 2 positions in Texarkana Message-ID: <20041108.174137.3692.0.mprice26@juno.com> Histonetters, We have 2 histotech positions to fill in Texarkana (Northeast,Texas) at Pathology Services of Texarkana. We also need a temporary histotech while we are looking. Please forward your resume via e-mail to me if you are interested. Thank you. Marsha Price ________________________________________________________________ Juno Platinum $9.95. Juno SpeedBand $14.95. Sign up for Juno Today at http://www.juno.com! Look for special offers at Best Buy stores. From tahseen <@t> brain.net.pk Mon Nov 8 19:57:20 2004 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Employ of the year Message-ID: <018501c4c5ff$73abfc80$972bfea9@m7c0y4> Dear All I am currently selecting employ of the year 2004 out of 50. If anyone has any ideas or experience or recommendations or can guide me to any appropriate software, literature I would be grateful. Thanks in advance Regards Muhammad Tahseen Laboratory Supervisor SKMCH & RC Lahore Pakistan. From lam-helen <@t> ctimail3.com Mon Nov 8 21:01:55 2004 From: lam-helen <@t> ctimail3.com (Helen Lam) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] ready-to-use commercial staining solutions Message-ID: <000801c4c608$79401f60$595bc3cb@computer> Dear all, Our lab is thinking about switching to ready-to-use commercial stains. = And I think there is no better place than the Histonet to gather = opinions form Histo.people, right? 1.. What are the pros and cons of using commercial staining solutions? 2.. Are there anybody out there already using it? Is it possible to = replace most if not all of the 'home-made' reagents you regularly use in = your stains by commercial products? If yes/no, why? 3.. Even if you are using commercial reagents, is it still necessary = to keep a small amount of dyes and chemicals in stock as a 'back-up', = just in case the supply of commercial reagents may become uncertain? 4.. How do you test the commerical reagents and prove that it works = satisfactorily on your sections / in your applications? 5.. Are the price , quality and shelf-life acceptable? (It would be = apprecaited if you could also indicate what brand of what staining = solution you are using.) 6.. Considering the price, quality and shelf-life of commerical = reagents, do you think they can satisfactorily replace 'home-made' = stains in our setting? We have 20-30 slides to do everyday by hand = (mainly alcian blue, PAS, a few renal and liver penals every week, some = requests for micro-organisms at times). By switching to commercial = reagents, we hope that we could reduce the amount or eliminate = altogether the need to keep a stock of dangerous chemicals and dye = powder. This in turn should save us some storage space, = money(especially for keeping stock of rarely used dyes or chemicals that = may go bad after years of standing) and man-hours in stain preparation. So what do you think? Any response to any of the above questions are = welcome. I look forward to hearing from you. Thanks in advance! Helen Lam Tuen Mun Hospital Hong Kong From zumbor <@t> email.cs.nsw.gov.au Mon Nov 8 23:58:49 2004 From: zumbor <@t> email.cs.nsw.gov.au (Rosalba) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Erado-sol Message-ID: <001801c4c621$2ed6dc70$1e7b4c98@dofm.cs.nsw.gov.au> Erado-sol is still available Try contacting the manufacturer for distributors Cambridge Diagnostic Products Inc 6880 N.W 17th Avenue Fort Lauderdale, FL 33309 PH : (954) 971-4040 FAX: (954) 975-5609 Rosalba Zumbo Laboratory Manager Histology Dept. Department of Forensic Medicine 42-50 Parramatta rd Glebe, 2037 Australia Ph: 61 2 85847842 Fax: 61 2 95664573 email: zumbor@email.cs.nsw.gov.au From tahseen <@t> brain.net.pk Tue Nov 9 03:25:41 2004 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Recommendations frozen section Message-ID: <011801c4c63e$168e19e0$972bfea9@m7c0y4> Dear All We are going to prepare a tutorial about the frozen section for our management staff. Does anybody have any documents or recommendations that they would be willing to share. Thanks in advance Tahseen Laboratory Supervisor SKMCH & RC Lahore, Pakistan. From lpwenk <@t> sbcglobal.net Tue Nov 9 04:08:18 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Employ of the year References: <018501c4c5ff$73abfc80$972bfea9@m7c0y4> Message-ID: <006601c4c644$09943480$0028d445@domainnotset.invalid> Maybe the National Society for Histotechnology awards web page has some ideas for you. http://www.nsh.org/membership/awardscholar.html Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Muhammad Tahseen" To: Sent: Monday, November 08, 2004 8:57 PM Subject: [Histonet] Employ of the year Dear All I am currently selecting employ of the year 2004 out of 50. If anyone has any ideas or experience or recommendations or can guide me to any appropriate software, literature I would be grateful. Thanks in advance Regards Muhammad Tahseen Laboratory Supervisor SKMCH & RC Lahore Pakistan. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Tue Nov 9 04:09:46 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Histological dissection of bone References: Message-ID: <006f01c4c644$3e0e0b00$0028d445@domainnotset.invalid> They have an "extender" which makes the clearance opening size larger, so bigger specimens can be passed through. Ask the company about it. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "DiCarlo, Margaret" To: "'Coaker, Terry'" ; Sent: Monday, November 08, 2004 11:46 AM Subject: RE: [Histonet] Histological dissection of bone > Terry, > > I also use a band saw for cutting bones of all different sizes, such as > femur, tibia, etc. People have raved about the Mar-Med Band Saw that costs > $810 but I don't think it is large enough for large sections of bone such as > a whole tibia or fibula that we have to cut up. It is an 18-pound tabletop > band saw with a diamond blade and is about the size of a microscope. I will > be interested to hear what everyone has to say. > > Peggy DiCarlo HT (ASCP) > Orthopedics Bone Lab > Buffalo General Hospital > 100 High St. > Buffalo, NY 14203 > 716-859-1293 > > > > -----Original Message----- > From: Coaker, Terry [mailto:Terry.Coaker@nuth.northy.nhs.uk] > Sent: Monday, November 08, 2004 10:30 > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] Histological dissection of bone > > > For many years we have "trimmed" bone samples on a band-saw to obtain slices > for decalcification. The equipment needs replacing. A risk assessment > indicated that putting a long bone through a band-saw in this way could lead > to someone getting hurt! Is there a safer way? Diamond saws are fine for > trimming small pieces but what about femur and humerus? How do people > handle them safely? > > > Thankyou > > Terry > > Terry Coaker > Histopathology Operations Manager > Cellular Pathology > Royal Victoria Infirmary > Queen Victoria Road > Newcastle upon Tyne > NE1 4LP > 0191 282 9120 > > _______________________________________________ > 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. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk Tue Nov 9 04:09:59 2004 From: JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk (JOHN PHILLIPS) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Immuno Artifact? Message-ID: <166A1E642B5B644DA694C08FD29D0ADC3E12B5@ztroy.new-tr.wales.nhs.uk> Make sure the sections are properly dehydrated and are wet before mountant is placed on them. If the sections are partially dried out then an artefact like this does appear. Good luck, John,Wrexham, Wales -----Original Message----- From: Bryan Hewlett [mailto:bhewlett@cogeco.ca] Sent: 08 November 2004 17:37 To: Marshall Terry Dr, Consultant Histopathologist; jason m; histonet@pathology.swmed.edu Subject: Re: [Histonet] Immuno Artifact? I agree, but it could also be water since the Becke lines do not appear quite wide enough for air. Bryan Hewlett, ART, MLT Consultant Technologist Quality Management Program-Laboratory Services Toronto, Ontario, Canada. ----- Original Message ----- From: "Marshall Terry Dr, Consultant Histopathologist" To: "jason m" ; Sent: Monday, November 08, 2004 12:12 PM Subject: RE: [Histonet] Immuno Artifact? That surely is entrapped air under the coverslip. 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: jason m [mailto:kosmicdog@hotmail.com] Sent: 08 November 2004 16:47 To: histonet@pathology.swmed.edu Subject: [Histonet] Immuno Artifact? Can anyone identify the cause of this black circle artifact? It is small black circles that look sort of like air bubbles only much too small which have been turning up on my immuno's this week. I have changed all solutions but still have the problem. Our lab uses toluene instead of xylenes, I am wondering if that is the problem. Also I use Tween-20 in my citrate buffer and I wonder if that could be the problem. At any rate I will be checking both of these but if anyone can help It will be appreciated immensely. I have sent a photograph of the artifact to the the list serv image archive (see link below) under the filename "artifact.jpg". http://www.histonet.org/site_images_frame.asp Thanks again, Jason _______________________________________________ Histonet mailing list Histonet@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 Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r rheolwr systemau wybod drwy ddefnyddio'r manylion isod. Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain Cymru. Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau???r Ddeddf Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru ar www.newalesnhstrust.org.uk This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the system manager using the details below. The contents of this email represent the views of the individual(s) named above and do not necessarily represent the views of the North East Wales NHS Trust. Please be aware that, under the terms of the Freedom of Information Act 2000, the North East Wales NHS Trust may be required to make public the content of any emails or correspondence received. For futher information on Freedom of Information, please refer to the North East Wales NHS Trust website at www.newalesnhstrust.org.uk For further assistance, please contact system.administrator@new-tr.wales.nhs.uk. From n.cragg <@t> epistem.co.uk Tue Nov 9 04:48:04 2004 From: n.cragg <@t> epistem.co.uk (ncragg) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Re: Immuno Artifact? Message-ID: <01C4C649.97DA12A0.n.cragg@epistem.co.uk> Is it 2 coverslips stuck together ? Done this myself!! Nicola Nicola Cragg BSc Epistem Ltd Manchester, UK From abright <@t> brightinstruments.com Tue Nov 9 05:49:54 2004 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Shandon Autosharp Message-ID: Yes we use an Autosharp 5 for sharpening tungsten carbide tipped knives, we use Kemet diamond compound Grade: 1-KD-C2 which is 1?, we purchase this, the lubricating oil, cleaning fluid & plates from http://www.kemet.co.uk and they have a distributor in the USA http://www.lapmaster.com Sorry I do not have a handbook for the Autosharp 5, but you can always email or phone me for advice. Best Regards Alan Bright Bright Instrument Co.Ltd. Cryostat & Microtome Divn. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Patty Lott [mailto:Patty.Lott@ORTHO.UAB.EDU] Sent: 08 November 2004 19:04 To: histonet@pathology.swmed.edu Subject: [Histonet] Shandon Autosharp Is anyone still using a Shandon Autosharp for sharpening tungsten carbide knives? I need a copy of the handbook, and I need to see what reagents (pastes, etc.) I would need to buy to put an old one which we found into service again! Patty Lott, Laboratory Supervisor Orthopaedic Research Laboratory Center for Metabolic Bone Disease Laboratory University of Alabama at Birmingham LHRB B37 0007 1919 7th Ave. South Birmingham, AL 35294 (205) 934-2007 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Luis.Chiriboga <@t> med.nyu.edu Tue Nov 9 07:52:49 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Histotechs on the net website Message-ID: Morning all, There used to be a website/page that listed (for those who registered) histotech's around the country. I believe it was called the "histotech's homepage" or the "histotech's portal". All the links that I had are dead and they send you to http://www.histology.to . Does anyone know what happened to the site? Does anyone know who the webmaster or administrator was? Thanks in advance L From DDittus787 <@t> aol.com Tue Nov 9 08:36:15 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Dumbing down the practical???] Message-ID: <61106667.262E00FC.0A1F969F@aol.com> I have been watching these conversations and have to say that it looks like the consesus is that things just ain't what they used to be. I agree no account is taken in when judging slides for automation versus manual, I agree that slides do not tell the whole story, I even take exception that the qihc exam is all slides and no written so does that mean you understand what a secondary is or quenching, or that many companies with automation have many people who passed hmmm? I take offense that almost all company people made up the IHC check list. I would love to be a reviewer and would love to see change.How about the rest of you???? dana From Michael.Rice <@t> holy-cross.com Tue Nov 9 08:32:37 2004 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] ready-to-use commercial staining solutions Message-ID: <3BC92F29BE821745AB15E04C98EE028D693643@HCH2KMAIL.holy-cross.com> Helen, For many years, (more than I care to remember)I prepared all of my own special stain reagents,but as time went on and companies began producing quality reagents I switched over and have never looked back. The company that I have used for many years is Poly Scientific in New York. They can be reached at (631)5860400 and will send you a catalogue. They will also produce custom reagents if you desire. Mike Rice Holy Cross Hospital Ft Lauderdale Florida -----Original Message----- From: Helen Lam [mailto:lam-helen@ctimail3.com] Sent: Monday, November 08, 2004 10:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] ready-to-use commercial staining solutions Dear all, Our lab is thinking about switching to ready-to-use commercial stains. And I think there is no better place than the Histonet to gather opinions form Histo.people, right? 1.. What are the pros and cons of using commercial staining solutions? 2.. Are there anybody out there already using it? Is it possible to replace most if not all of the 'home-made' reagents you regularly use in your stains by commercial products? If yes/no, why? 3.. Even if you are using commercial reagents, is it still necessary to keep a small amount of dyes and chemicals in stock as a 'back-up', just in case the supply of commercial reagents may become uncertain? 4.. How do you test the commerical reagents and prove that it works satisfactorily on your sections / in your applications? 5.. Are the price , quality and shelf-life acceptable? (It would be apprecaited if you could also indicate what brand of what staining solution you are using.) 6.. Considering the price, quality and shelf-life of commerical reagents, do you think they can satisfactorily replace 'home-made' stains in our setting? We have 20-30 slides to do everyday by hand (mainly alcian blue, PAS, a few renal and liver penals every week, some requests for micro-organisms at times). By switching to commercial reagents, we hope that we could reduce the amount or eliminate altogether the need to keep a stock of dangerous chemicals and dye powder. This in turn should save us some storage space, money(especially for keeping stock of rarely used dyes or chemicals that may go bad after years of standing) and man-hours in stain preparation. So what do you think? Any response to any of the above questions are welcome. I look forward to hearing from you. Thanks in advance! Helen Lam Tuen Mun Hospital Hong Kong _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. 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 jo-ann-e.bader <@t> staff.mcgill.ca Tue Nov 9 08:50:31 2004 From: jo-ann-e.bader <@t> staff.mcgill.ca (jo-ann-e.bader@staff.mcgill.ca) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] NFB vs. Paraformaldehyde Message-ID: <1100011831.4190d937e55d2@webmail.mcgill.ca> Good Morning Histonetters, I would like to get an idea from those histotechs that work with mouse tissue if they prefer NFB or Paraformaldehyde as a fixative for routine histology. I work for a group of 6 researchers, most of who do mouse work. Their students do the necropsies, remove and block the tissue, make up their own paraformaldehyde, fix and submit the cassettes to me from processing and staining. I would like everyone to change to NFB to standarize the fixation. I can guarentee that no one makes Paraformaldehyde the same. Some make it the same day, some the day before, 2-3 days before etc. I am meeting with the big bosses this afternoon and would like some ammunition if possible. Thanks for the assistance. Jo-Ann From jim <@t> hteqa.co.uk Tue Nov 9 09:39:49 2004 From: jim <@t> hteqa.co.uk (Jim Elsam) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] Histological dissection of bone Message-ID: Hi Terry In the days when I had a proper job at the Christie, we too used a diamond saw for bone. We got ours from Manchester Minerals www.manchesterminerals.co.uk and I found them very helpful and not overly expensive. Diamond saws, while great for hard stuff, are useless for soft tissue, fingers included, so they are very safe. Indeed they are so safe that you may need to incise any cartilage around the bone before you can get started. Best wishes ~~~~~~~~~~~~~~~~~~~~~~ Jim Elsam, HTEQA Services Ltd From cfavara <@t> niaid.nih.gov Tue Nov 9 09:36:22 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] IP-10,D18 & D13 Message-ID: Anyone with experience using these antibodies on formalin fixed paraffin embedded tissue? IP-10 = Interferon Inducible Protein 10 CXC chemokine D18 and D13 are antibodies prion protein epitopes. 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 ploykasek <@t> phenopath.com Tue Nov 9 09:59:43 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:16 2005 Subject: [Histonet] slide stainers and labelers In-Reply-To: <20041108201338.83244.qmail@web60610.mail.yahoo.com> Message-ID: Jennifer - We are currently doing an evaluation of the Sakura slide printer. I should give you some background on how we operate. We are not a routine histology lab - we're not cutting only 1-3 H&E's on blocks. We are a reference IHC/ISH laboratory, typically we cut 6-15 unstained slides & 1 H&E on blocks. We receive the majority of our blocks by Fed EX, so they arrive en masse mid-morning. We were having 3-4 people write slides for at least an hour. The slide printer needs just 1 person, and is much faster doing all the slides. Legibility is no longer a problem. Even the doctors have remarked on how easy & quick it is to read the slides with the uniform printing. It took us awhile to get the software configured with our programs the way we wanted it, but this was time well spent. We're still trying to get it to print out paper reports of what we're inputting (so we don't have to hand write logs). We have not tried hooking it up to our LIS yet. Of course, this machine does not prevent errors. You can type in the incorrect number, pick up tissue on the wrong slides, etc... I definitely have not seen any increase in errors. I'm truly sold on the machine due to less tech time spent writing slides, improved legibility, quickness of printing. Now I've just got to write new SOPs!!! Let me know if you'd like more info. Patti Loykasek PhenoPath Laboratories Seattle, WA > I'm looking for any info on slide stainers and slide lablers. Our lab is > looking to purchase one of each. All responses will be greatly appreciated. > > Thanks to everyone for helping me out whether it is now or in the past. You > guys ar egreat and I love this forum! > > > --------------------------------- > Do you Yahoo!? > Check out the new Yahoo! Front Page. www.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PatPatterson <@t> mhd.com Tue Nov 9 10:01:04 2004 From: PatPatterson <@t> mhd.com (Patterson, Pat) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] DiffQuik Question again Message-ID: <293C7C19EFF7D611AE1A0002A53F81140CB43B48@omega.mhd.com> Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. From Luis.Chiriboga <@t> med.nyu.edu Tue Nov 9 10:09:37 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] IP-10,D18 & D13 In-Reply-To: Message-ID: Hi Cynthia I have been playing around with IP10 from R&D systems (AF-266-NA). It's goat anti-human antibody that have used for ELI spot. As with all of the anti-chemokine antibodies, it's difficult to get consistent and reproducible results in FF-PE. I haven't gotten some results but these are preliminary. Let me know if you need more info... Hope this helps Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Favara, Cynthia (NIH/NIAID) Sent: Tuesday, November 09, 2004 10:36 AM To: HistoNet Server (HistoNet Server) Subject: [Histonet] IP-10,D18 & D13 Anyone with experience using these antibodies on formalin fixed paraffin embedded tissue? IP-10 = Interferon Inducible Protein 10 CXC chemokine D18 and D13 are antibodies prion protein epitopes. 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 mcauliff <@t> umdnj.edu Tue Nov 9 13:20:13 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] NFB vs. Paraformaldehyde In-Reply-To: <1100011831.4190d937e55d2@webmail.mcgill.ca> References: <1100011831.4190d937e55d2@webmail.mcgill.ca> Message-ID: <4191186D.3070407@umdnj.edu> Hi Jo-Ann: It may not matter that the paraformaldehyde is a few days or even a few weeks old, seems that the "paraformaldehyde must be made fresh " dogma is a myth (this was published recently, I just wish I knew where). NBF from a bottle is a little easier and the little bit of methanol used as a perservative does not make a difference at the LM level. Getting people to believe that is another matter! On the other hand, the students do need to get their hands wet (so to speak) in the lab so that when they have their own lab they can train others. And it probably does not matter if each student makes his/her fix a bit differently. If they make a big mistake, then it is their problem. By the way, I think they should be doing their own processing, sectioning and staining. It really is part of their training. How else will they learn? Geoff jo-ann-e.bader@staff.mcgill.ca wrote: >Good Morning Histonetters, > >I would like to get an idea from those histotechs that work with mouse tissue if >they prefer NFB or Paraformaldehyde as a fixative for routine histology. > >I work for a group of 6 researchers, most of who do mouse work. Their students >do the necropsies, remove and block the tissue, make up their own >paraformaldehyde, fix and submit the cassettes to me from processing and >staining. > >I would like everyone to change to NFB to standarize the fixation. I can >guarentee that no one makes Paraformaldehyde the same. Some make it the same >day, some the day before, 2-3 days before etc. > >I am meeting with the big bosses this afternoon and would like some ammunition >if possible. > >Thanks for the assistance. > >Jo-Ann > >_______________________________________________ >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 juan.gutierrez <@t> christushealth.org Tue Nov 9 10:33:12 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] slide stainers and lablers Message-ID: We do not have labelers, but we recently purchased a stainer. I'm assuming you are asking about H&E stainers. We purchased the Microm HMS760X. It is a very good instrument with the ability to run different programs at the same time with continuous loading. It comes with a computer that shows you where every rack is and all sorts of information about time, next step and age of reagents. We also tested the new Surgipath stainer and found it to be very good also, providing some of the same information as the Microm in a smaller package (the Microm stainer is quite large, so space is an issue). I have also used the Thermo Gemini and thought it to be a nice system except for some software glitches and structural defects. The last time I talked to a sales rep. he assured me that they had been fixed. Last, I have also used the Sakura stainer and found it to be very good, but also quite large and not as informative as the three previous ones. My recommendation is try as many as you can and don't rush your decision because once you spend that kind of money on a piece of equipment you're expected to keep it for many years to come. If you were looking for immuno stainer my one choice is the Ventana Benchmark XT. 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: Jennifer Sipes [mailto:jengirl1014@yahoo.com] Sent: Monday, November 08, 2004 2:14 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] slide stainers and lablers I'm looking for any info on slide stainers and slide lablers. Our lab is looking to purchase one of each. All responses will be greatly appreciated. Thanks to everyone for helping me out whether it is now or in the past. You guys ar egreat and I love this forum! --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Tue Nov 9 10:38:41 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] NFB vs. Paraformaldehyde References: <1100011831.4190d937e55d2@webmail.mcgill.ca> Message-ID: <4190F291.E2D88F00@uwo.ca> This question comes up frequently on Histonet. The simple fact is that paraformaldehyde is an insoluble solid polymer that depolymerizes when heated in buffered water. The solution you end up with contains no paraformaldehyde at all, and is almost identical to a buffered formaldehyde solution made from formalin (which is a 37% solution of formaldehyde). Formalin consists largely of smaller polymers, which break down on dilution with water or buffer. A fixative made from formalin contains about 1% methanol, which is not present when paraformaldehyde is the source of the formaldehyde. If a formalin stock is very old (years) there will be more methanol, and also some formate ions. Small amounts of methanol and formate do not affect the fixative properties. A properly formulated buffered formaldehyde solution, made from formalin, is good even for subsequent electron microscopy. It is more trouble to make the fixative from paraformaldehyde. Remember, nobody has ever fixed a specimen in paraformaldehyde; this substance exists only in the solid state and is insoluble. Old bottles of formalin often contain a white precipitate in the bottom; this is paraformaldehyde. Methanol (about 10%) is added to formalin to retard the formation of this high polymer. For longer essays on this topic, look up: http://publish.uwo.ca/~jkiernan/formglut.htm Researchers these days frequently do not know the basics of fixation, staining or microscopy. For them, there is: http://publish.uwo.ca/~jkiernan/bookfind.htm#MOLB -- ------------------------------- 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 _______________________________ jo-ann-e.bader@staff.mcgill.ca wrote: > > Good Morning Histonetters, > > I would like to get an idea from those histotechs that work with mouse tissue if > they prefer NFB or Paraformaldehyde as a fixative for routine histology. > > I work for a group of 6 researchers, most of who do mouse work. Their students > do the necropsies, remove and block the tissue, make up their own > paraformaldehyde, fix and submit the cassettes to me from processing and > staining. > > I would like everyone to change to NFB to standarize the fixation. I can > guarentee that no one makes Paraformaldehyde the same. Some make it the same > day, some the day before, 2-3 days before etc. > > I am meeting with the big bosses this afternoon and would like some ammunition > if possible. > > Thanks for the assistance. > > Jo-Ann > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Tue Nov 9 10:41:41 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] NFB vs. Paraformaldehyde Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA257F6B0@usca0082k08.labvision.apogent.com> Jo-Ann, Way back in 1973 (ref below) Frieda Carson showed that when comparing quality of fixation between formaldehyde and paraformaldehyde, the buffer is the critical factor, not the source of the formalin. Paraformaldehyde is the solid form of formaldehyde. Once dissolved in water there is no difference between formalin from formaldehyde gas dissolved in water (what most of us use) and formalin made from paraformaldehyde. While commercial formalin has methanol included to prevent polymerization of the formaldehyde, that does not affect the fixative properties. So, in short, it would be much better to have the students pay attention to the buffers that are used than the source of the formalin. Of course, making the formalin will be much easier with concentrate than having to dissolve paraformaldehyde. In fact, a good class experiment would be to compare the two with the same buffers. Carson, FL, Lynn JA, Martin JN: Formalin fixation for electron microscopy: A re-evaluation. Am J Clin Pathol 59:365, 1973. Carson FL, Lynn JA, Martin JN: Ultrastuctural effect of various buffers, osmolality, and temperature on paraformaldehyde fixation of the formed elements of blood and bone marrow. Texas Rep Miol Med 30: 125, 1972 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: jo-ann-e.bader@staff.mcgill.ca [mailto:jo-ann-e.bader@staff.mcgill.ca] Sent: Tuesday, November 09, 2004 6:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] NFB vs. Paraformaldehyde Good Morning Histonetters, I would like to get an idea from those histotechs that work with mouse tissue if they prefer NFB or Paraformaldehyde as a fixative for routine histology. I work for a group of 6 researchers, most of who do mouse work. Their students do the necropsies, remove and block the tissue, make up their own paraformaldehyde, fix and submit the cassettes to me from processing and staining. I would like everyone to change to NFB to standarize the fixation. I can guarentee that no one makes Paraformaldehyde the same. Some make it the same day, some the day before, 2-3 days before etc. I am meeting with the big bosses this afternoon and would like some ammunition if possible. Thanks for the assistance. Jo-Ann _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Tue Nov 9 10:56:31 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Merkel cell staining References: <4188B405.A7723577@cormack.uct.ac.za> Message-ID: <4190F6BF.BAB39CB9@uwo.ca> I don't know if this helps, but there is a Merkel's fixative dating from 1870. Mix equal volumes of 1:400 chromium trioxide and 1:400 platinic chloride (presumably = H2PtCl6). Immerse objects (presumably very small or thin) for several hours to a few days. "After washing out with alcohol, objects stain excellently notwithstanding the admixture of chromic acid." (A. Bolles Lee 1885. The Microtomist's Vade Mecum. London: Churchill. pp 23 and 158-159. [Facsimile edition published in 1987 by Science Heritage Ltd, Lincolnwood, IL] Quinacrine has been used as a vital fluorochrome for Merkel cells in amphibian and rat epidermis. Crowe & Whitear 1978. Cell Tiss. Res. 190:273-283. Nurse et al 1983. Cell Tiss. Res. 228:511-524. I've not tried any of this myself! -- ------------------------------- 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 _______________________________ Marshall wrote: > > Hi all > I am looking to demonstrate Merkel cells in skin. I can not find any > reference in my textbooks to their demonstration although I have not > used the internet. Also I have tried silver stains for demoing free > nerve endings in skin with not much success. The immunocytochemistry for > neuro filament protein was a bit better but not great. Is there anyway I > can improve on this? > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ From sluhisto <@t> yahoo.com Tue Nov 9 10:56:14 2004 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Necrotic tissue stain Message-ID: <20041109165614.62701.qmail@web51010.mail.yahoo.com> Hello All: I have a research investigator who is inquiring about a procedure to identify necrotic cells. Apparently these cells are not undergoing apoptosis but rather are appearing swollen, with swollen nuclei and mitochondria. He believes that they are necrotic but is looking for some kind of stain or procedure that will distinguish with some specificity these cells from the normal ones. Any thoughts or help would be greatly appreciated. Susan --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com From pruegg <@t> ihctech.net Tue Nov 9 11:35:07 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Histotechs on the net website In-Reply-To: Message-ID: <000e01c4c682$747d18a0$83020a0a@IHCTech> That was Steve Slap who is no longer with us. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Luis Chiriboga Sent: Tuesday, November 09, 2004 6:53 AM To: Histonet Subject: [Histonet] Histotechs on the net website Morning all, There used to be a website/page that listed (for those who registered) histotech's around the country. I believe it was called the "histotech's homepage" or the "histotech's portal". All the links that I had are dead and they send you to http://www.histology.to . Does anyone know what happened to the site? Does anyone know who the webmaster or administrator was? Thanks in advance L _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Tue Nov 9 11:41:07 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] NFB vs. Paraformaldehyde Message-ID: <566FB0B522443D43AF02D2ADBE35A6F001008053@UTHEVS3.mail.uthouston.edu> Actually not to detract from the references below but a lot of the work in this area was carried out by Karlsson and Schultze in 1965. They were working with different fixatives for studying the electron microscopy of the brain and found that indeed the buffer made a significant difference to the final image. They also compared immersion via perfusion fixation. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Tuesday, November 09, 2004 10:42 AM To: 'jo-ann-e.bader@staff.mcgill.ca'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] NFB vs. Paraformaldehyde Jo-Ann, Way back in 1973 (ref below) Frieda Carson showed that when comparing quality of fixation between formaldehyde and paraformaldehyde, the buffer is the critical factor, not the source of the formalin. Paraformaldehyde is the solid form of formaldehyde. Once dissolved in water there is no difference between formalin from formaldehyde gas dissolved in water (what most of us use) and formalin made from paraformaldehyde. While commercial formalin has methanol included to prevent polymerization of the formaldehyde, that does not affect the fixative properties. So, in short, it would be much better to have the students pay attention to the buffers that are used than the source of the formalin. Of course, making the formalin will be much easier with concentrate than having to dissolve paraformaldehyde. In fact, a good class experiment would be to compare the two with the same buffers. Carson, FL, Lynn JA, Martin JN: Formalin fixation for electron microscopy: A re-evaluation. Am J Clin Pathol 59:365, 1973. Carson FL, Lynn JA, Martin JN: Ultrastuctural effect of various buffers, osmolality, and temperature on paraformaldehyde fixation of the formed elements of blood and bone marrow. Texas Rep Miol Med 30: 125, 1972 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: jo-ann-e.bader@staff.mcgill.ca [mailto:jo-ann-e.bader@staff.mcgill.ca] Sent: Tuesday, November 09, 2004 6:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] NFB vs. Paraformaldehyde Good Morning Histonetters, I would like to get an idea from those histotechs that work with mouse tissue if they prefer NFB or Paraformaldehyde as a fixative for routine histology. I work for a group of 6 researchers, most of who do mouse work. Their students do the necropsies, remove and block the tissue, make up their own paraformaldehyde, fix and submit the cassettes to me from processing and staining. I would like everyone to change to NFB to standarize the fixation. I can guarentee that no one makes Paraformaldehyde the same. Some make it the same day, some the day before, 2-3 days before etc. I am meeting with the big bosses this afternoon and would like some ammunition if possible. Thanks for the assistance. Jo-Ann _______________________________________________ Histonet mailing list Histonet@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 billingconsultants <@t> yahoo.com Tue Nov 9 11:58:07 2004 From: billingconsultants <@t> yahoo.com (Caldwell) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Antibody Help microphthalmia and CD34 Message-ID: <20041109175807.49675.qmail@web54209.mail.yahoo.com> Hi Everyone, I am looking for a good antibody for CD34 and microphthalmia-associated transcription factor. We have tried a couple of vendors for the CD34, and the pathologist hasn't been happy with them. Any suggestions would be greatly appreciated. Thank you in advance for your help. Louri Roberts-Caldwell --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com From Luis.Chiriboga <@t> med.nyu.edu Tue Nov 9 12:11:54 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Histotechs on the net website In-Reply-To: <000e01c4c682$747d18a0$83020a0a@IHCTech> Message-ID: my apologies to all, for some reason I never made the association, I guess because I had never met him. perhaps a downside to the internet world...... Thank you patsy, Will the info still be available through some other site, it would be even a greater shame to lose that great resource as well? Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Tuesday, November 09, 2004 12:35 PM To: 'Luis Chiriboga'; 'Histonet' Subject: RE: [Histonet] Histotechs on the net website That was Steve Slap who is no longer with us. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Luis Chiriboga Sent: Tuesday, November 09, 2004 6:53 AM To: Histonet Subject: [Histonet] Histotechs on the net website Morning all, There used to be a website/page that listed (for those who registered) histotech's around the country. I believe it was called the "histotech's homepage" or the "histotech's portal". All the links that I had are dead and they send you to http://www.histology.to . Does anyone know what happened to the site? Does anyone know who the webmaster or administrator was? Thanks in advance L _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Tue Nov 9 12:06:37 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Re: PCNA Ab Message-ID: Wen, DakoCytomation Monoclonal Mouse PCNA, Clone PC10 (M 0879) works well in mouse tissues @ 1:200 with antigen unmasking using Citrate buffer pH 6.0. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From cfavara <@t> niaid.nih.gov Tue Nov 9 12:07:51 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] NFB vs. Paraformaldehyde Message-ID: To All, I also work with many different researchers and they all have their own idea as to what fixative should be used. My solution: I provide NBF freshly made from 37% formaldehyde if requested. I make small quantities as that seems to be to most researchers liking. I have a SOP for paraformaldehyde if a researcher chooses to go to the trouble. I know very few of them take the time to make this properly and you can only imagine the variation. I do not make paraformaldehyde because I have never been able to convince myself it makes any difference for the stains I do. Most of the staining variability I see is a result of a researcher insisting on their own brand of fixative without any normal controls fixed and processed the same way. Just some ramblings! Creative procrastination! 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: Barry R Rittman [mailto:Barry.R.Rittman@uth.tmc.edu] Sent: Tuesday, November 09, 2004 10:41 AM To: histonet Subject: RE: [Histonet] NFB vs. Paraformaldehyde Actually not to detract from the references below but a lot of the work in this area was carried out by Karlsson and Schultze in 1965. They were working with different fixatives for studying the electron microscopy of the brain and found that indeed the buffer made a significant difference to the final image. They also compared immersion via perfusion fixation. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Tuesday, November 09, 2004 10:42 AM To: 'jo-ann-e.bader@staff.mcgill.ca'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] NFB vs. Paraformaldehyde Jo-Ann, Way back in 1973 (ref below) Frieda Carson showed that when comparing quality of fixation between formaldehyde and paraformaldehyde, the buffer is the critical factor, not the source of the formalin. Paraformaldehyde is the solid form of formaldehyde. Once dissolved in water there is no difference between formalin from formaldehyde gas dissolved in water (what most of us use) and formalin made from paraformaldehyde. While commercial formalin has methanol included to prevent polymerization of the formaldehyde, that does not affect the fixative properties. So, in short, it would be much better to have the students pay attention to the buffers that are used than the source of the formalin. Of course, making the formalin will be much easier with concentrate than having to dissolve paraformaldehyde. In fact, a good class experiment would be to compare the two with the same buffers. Carson, FL, Lynn JA, Martin JN: Formalin fixation for electron microscopy: A re-evaluation. Am J Clin Pathol 59:365, 1973. Carson FL, Lynn JA, Martin JN: Ultrastuctural effect of various buffers, osmolality, and temperature on paraformaldehyde fixation of the formed elements of blood and bone marrow. Texas Rep Miol Med 30: 125, 1972 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: jo-ann-e.bader@staff.mcgill.ca [mailto:jo-ann-e.bader@staff.mcgill.ca] Sent: Tuesday, November 09, 2004 6:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] NFB vs. Paraformaldehyde Good Morning Histonetters, I would like to get an idea from those histotechs that work with mouse tissue if they prefer NFB or Paraformaldehyde as a fixative for routine histology. I work for a group of 6 researchers, most of who do mouse work. Their students do the necropsies, remove and block the tissue, make up their own paraformaldehyde, fix and submit the cassettes to me from processing and staining. I would like everyone to change to NFB to standarize the fixation. I can guarentee that no one makes Paraformaldehyde the same. Some make it the same day, some the day before, 2-3 days before etc. I am meeting with the big bosses this afternoon and would like some ammunition if possible. Thanks for the assistance. Jo-Ann _______________________________________________ Histonet mailing list Histonet@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 juan.gutierrez <@t> christushealth.org Tue Nov 9 12:29:38 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] DiffQuik Question again Message-ID: We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Tue Nov 9 12:33:37 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] slide stainers and labelers References: Message-ID: <005b01c4c68a$a0fdf180$eeeea8c0@server> Dear Patti I would like to hear, if you have to mark the slides with the histotechs initials, who cuts. We label the slides by hand just before cutting. With the handwriting we allways know the originator of the slide. Our laboratory is going to certificate and there is the request to follow the probe's way. I would appreciate a slide labeler, because my handwriting is awful .... Gudrun ----- Original Message ----- From: "Patti Loykasek" To: "Jennifer Sipes" ; "histonet" Sent: Tuesday, November 09, 2004 4:59 PM Subject: Re: [Histonet] slide stainers and labelers > Jennifer - We are currently doing an evaluation of the Sakura slide printer. > I should give you some background on how we operate. We are not a routine > histology lab - we're not cutting only 1-3 H&E's on blocks. We are a > reference IHC/ISH laboratory, typically we cut 6-15 unstained slides & 1 H&E > on blocks. We receive the majority of our blocks by Fed EX, so they arrive > en masse mid-morning. We were having 3-4 people write slides for at least an > hour. The slide printer needs just 1 person, and is much faster doing all > the slides. Legibility is no longer a problem. Even the doctors have > remarked on how easy & quick it is to read the slides with the uniform > printing. It took us awhile to get the software configured with our programs > the way we wanted it, but this was time well spent. We're still trying to > get it to print out paper reports of what we're inputting (so we don't have > to hand write logs). We have not tried hooking it up to our LIS yet. > Of course, this machine does not prevent errors. You can type in the > incorrect number, pick up tissue on the wrong slides, etc... I definitely > have not seen any increase in errors. > I'm truly sold on the machine due to less tech time spent writing slides, > improved legibility, quickness of printing. > Now I've just got to write new SOPs!!! > Let me know if you'd like more info. > > Patti Loykasek > PhenoPath Laboratories > Seattle, WA > > > > > > I'm looking for any info on slide stainers and slide lablers. Our lab is > > looking to purchase one of each. All responses will be greatly appreciated. > > > > Thanks to everyone for helping me out whether it is now or in the past. You > > guys ar egreat and I love this forum! > > > > > > --------------------------------- > > Do you Yahoo!? > > Check out the new Yahoo! Front Page. www.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 sharon.osborn <@t> dnax.org Tue Nov 9 12:45:56 2004 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] RE: cassette and slide labelers Message-ID: <29B25753F6B1D51196110002A589D44401A178AB@PALMSG30.us.schp.com> Jennifer, This lab have been using the TBS cassette and slide labelers for several years. I first used them in 1994. They etch the slides and burn w/black ink onto the cassettes. There are problems with them that require a lot of attention. In addition, the glass and paint dust on the slides must be washed off. A lot of ours go to IHC and that is the main reason for washing off the dust. Yes, we have the 'dust catcher' attachment on it. If service is needed, we must make arrangements with TBS and rent a loaner, return ours for repair, etc. Recently, we requested demonstrations of the Shandon, Surgipath, Leica, Sakura instruments. Our requirements were it was to hook into our LIS system. We spoke with TBS concerning their newly released totally different labeler. It is still too new to have all the problems worked out and since there is not a service rep on the West Coast, service would be a huge issue. 1. Surgipath--it is a small one slide at a time instrument; not suited for mass production nor will it integrate with an LIS. Can rub off the ink. 2. Shandon--slide labeler is exactly the same as the TBS. The cassette labeler has a carousel configuration rather than the slide configuration TBS has. Both still operate same as the TBS. Ink will rub, scratch off with some effort. In practise, it takes only certain types of slides (both TBS and Shandon) 3. Sakura--it turned out to be same as the Leica. It was developed with some type of joint agreement that is no longer in effect. (ac/c to Sakura representative) 4. Leica--Both the cassette labeler and slide write take up more bench top space than the TBS or Shandon. However, it is space well worth taking. It will take casettes with lids on it or without. It will take any type of slides. It will more easily hook into our LIS system such that researchers can program from their desks the cassettes to print. We can remotely program it to print slides. Bottom line--We ordered the Leica because we received a better price quote from the Leica representative than from the Sakura representative. Plus, we have had excellent service on our other Leica products. I do agree that you want to demo as many of these big ticket items as you can due to the longevity of relationship with the piece of equipment. Sharon Osborn DNAX Palo Alto, CA ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From MadaryJ <@t> MedImmune.com Tue Nov 9 13:00:49 2004 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] slide labelers Message-ID: <83899F0EC7671543B305FB5694024DE60A1B458A@medimmune4.medimmune.com> We purchased a Lamb Microwriter 1-866-289-5262. It seems to do very well for the slides and they will also give you a decent proce on the slides as well. From MadaryJ <@t> MedImmune.com Tue Nov 9 13:09:18 2004 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] NBF for Mouse tissues Message-ID: <83899F0EC7671543B305FB5694024DE60A1B458C@medimmune4.medimmune.com> Paraformaldehyde would not really make it routine for you since it is not as easy as NBF. Paraformaldehyde after it is made up is almost the same as the 10% NBF as far as routine fixation is concerned from my experience. We have been using 10% NBF and it works great. From Rcartun <@t> harthosp.org Tue Nov 9 13:17:16 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] DiffQuik Question again Message-ID: Dear Juan: Do all these patients get billed for the Giemsa histochemical stain? 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 >>> "GUTIERREZ, JUAN" 11/09/04 01:29PM >>> We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Tue Nov 9 13:34:12 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Histotechs on the net website In-Reply-To: Message-ID: <000401c4c693$175a42e0$83020a0a@IHCTech> I don't know, Peggy Wenk was helping with that I thought??? Patsy -----Original Message----- From: Luis Chiriboga [mailto:Luis.Chiriboga@med.nyu.edu] Sent: Tuesday, November 09, 2004 11:12 AM To: Patsy Ruegg; 'Histonet' Subject: RE: [Histonet] Histotechs on the net website my apologies to all, for some reason I never made the association, I guess because I had never met him. perhaps a downside to the internet world...... Thank you patsy, Will the info still be available through some other site, it would be even a greater shame to lose that great resource as well? Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Tuesday, November 09, 2004 12:35 PM To: 'Luis Chiriboga'; 'Histonet' Subject: RE: [Histonet] Histotechs on the net website That was Steve Slap who is no longer with us. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Luis Chiriboga Sent: Tuesday, November 09, 2004 6:53 AM To: Histonet Subject: [Histonet] Histotechs on the net website Morning all, There used to be a website/page that listed (for those who registered) histotech's around the country. I believe it was called the "histotech's homepage" or the "histotech's portal". All the links that I had are dead and they send you to http://www.histology.to . Does anyone know what happened to the site? Does anyone know who the webmaster or administrator was? Thanks in advance L _______________________________________________ Histonet mailing list Histonet@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 Tue Nov 9 13:37:31 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] DiffQuik Question again Message-ID: <83AACDB0810528418AA106F9AE9B7F7E5074D7@sjhaexc02.sjha.org> We do Genta on all gastric bxs and then if they are not necessary we credit the charge. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Tuesday, November 09, 2004 2:17 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again Dear Juan: Do all these patients get billed for the Giemsa histochemical stain? 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 >>> "GUTIERREZ, JUAN" 11/09/04 01:29PM >>> We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@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 ** 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 Tue Nov 9 13:51:49 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] DiffQuik Question again Message-ID: Yes, my gross room tech bills for it automatically. 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: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Tuesday, November 09, 2004 1:17 PM To: GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again Dear Juan: Do all these patients get billed for the Giemsa histochemical stain? 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 >>> "GUTIERREZ, JUAN" 11/09/04 01:29PM >>> We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@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 Tue Nov 9 13:56:30 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] DiffQuik Question again Message-ID: Who determines when it's not necessary? If a stain is negative it does not necessarily mean it was not warranted. The stain is done to figure out if the bugs are there or not. The clinician needs to know either way. Because even if it's a tumor, did the bugs have a role in the process? 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: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Tuesday, November 09, 2004 1:38 PM To: Richard Cartun; GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again We do Genta on all gastric bxs and then if they are not necessary we credit the charge. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Tuesday, November 09, 2004 2:17 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again Dear Juan: Do all these patients get billed for the Giemsa histochemical stain? 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 >>> "GUTIERREZ, JUAN" 11/09/04 01:29PM >>> We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@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 ** 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 darrenj <@t> medica.co.nz Tue Nov 9 14:08:15 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] slide stainers and labellers In-Reply-To: <20041108201338.83244.qmail@web60610.mail.yahoo.com> Message-ID: <000401c4c697$d974d350$c864a8c0@medica.co.nz> Hi Jennifer, In regards to slide labellers the Shandon model is very good. The writing is etched on rather than printed. Shandon also are in the process of releasing a laser slide writer. This is a beautiful machine which produces slides of fantastic quality. Again the information is etched on using laser rather than being printed, the clarity and detail is something to behold. (It even prints 2D barcodes). Personally I would be hesitant to use anything which uses ink. I am sure the developers of the ink printers have done their homework on the ink properties but I just cringe when it comes to ink, alcohol, xylene and other solvents. As a side issue the Shandon cassette writers burn the data onto the cassette so that isn't going anywhere either (unless you particularly want to scratch it off). As for stainers the Autostainer RSt is great - see link http://www.vision-bio.com/product_autostainer.html I have used this for many years in various labs and it is bullet-proof. It can store up to 15 programmes and can stain up to 200 slides per hour(protocol dependant). Hope this is useful. Thanks Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jennifer Sipes Sent: Tuesday, 9 November 2004 9:14 a.m. To: histonet@lists.utsouthwestern.edu Subject: [Histonet] slide stainers and lablers I'm looking for any info on slide stainers and slide lablers. Our lab is looking to purchase one of each. All responses will be greatly appreciated. Thanks to everyone for helping me out whether it is now or in the past. You guys ar egreat and I love this forum! --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BWinters <@t> NCH.ORG Tue Nov 9 14:04:17 2004 From: BWinters <@t> NCH.ORG (Winters, Bert) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] SAKURA XPRESS Message-ID: <270614B321ACB44D8C1D91F4F921FDC3627786@NCH01EX02.nch.org> For all sakura xpress users: We are trying to justify the cost of this equipment. If anyone can help with data showing reduction in length of stay or any other justification facts that will help me sell this piece of equipment to administration I would be greatly appreciate it. Bert Winters Northwest community hospital Arlington heights, Ill. 60005 E-mail: BWINTERS@NCH.ORG ******************* PLEASE NOTE ******************* This E-Mail/telefax message and any documents accompanying this transmission may contain information that is privileged, confidential, and/or exempt from disclosure under applicable law and is intended solely for the addressee(s) named above. If you are not the intended addressee/recipient, you are hereby notified that any use of, disclosure, copying, distribution, or reliance on the contents of this E-Mail/telefax information is strictly prohibited and may result in legal action against you. Please reply to the sender advising of the error in transmission and immediately delete/destroy the message and any accompanying documents. Thank you. From GoodwinD <@t> pahosp.com Tue Nov 9 14:22:35 2004 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Full time position available, Pennsylvania Hospital, Phila, PA. Message-ID: <992899E9EC268548AB8DDE246AF88473055F4BBE@PAHEX01.uphs.upenn.edu> Histotechnologist, Full Time As the nation's first hospital, our legacy of quality healthcare started over 250 years ago. Recognized as a leader by U.S. News & World Report, Pennsylvania Hospital is a 534-bed acute care facility located in historic Society Hill. We offer an EXCELLENT salary and benefits package. For a complete position description and to apply online, please visit our Web site at http:// www.pennhealth.com/jobs. AA/EOE M/F/D/V, or e-mail resume with cover letter to goodwind@pahosp.com . University of Pennsylvania Health System From sluhisto <@t> yahoo.com Tue Nov 9 14:46:10 2004 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Histotechnologist position Message-ID: <20041109204610.45932.qmail@web51010.mail.yahoo.com> St. Louis University School of Medicine, St. Louis, Mo. has a full time day opening for a registered histotechnologist. If interested, please send CV to Susan Hay-Antoff, 1402 So. Grand Blvd, Room 464, St. Louis, Mo. 63104. --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com From asmith <@t> mail.barry.edu Tue Nov 9 14:49:47 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Immuno Artifact? Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3C33@exchsrv01.barrynet.barry.edu> It looks to me like the sort of air bubbles one gets when the tissue starts to dry before the coverslip is lowered into place. I suspect that toluene is the source of your problem: toluene dries much faster than xylene. 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 jason m Sent: Monday, November 08, 2004 11:47 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Immuno Artifact? Can anyone identify the cause of this black circle artifact? It is small black circles that look sort of like air bubbles only much too small which have been turning up on my immuno's this week. I have changed all solutions but still have the problem. Our lab uses toluene instead of xylenes, I am wondering if that is the problem. Also I use Tween-20 in my citrate buffer and I wonder if that could be the problem. At any rate I will be checking both of these but if anyone can help It will be appreciated immensely. I have sent a photograph of the artifact to the the list serv image archive (see link below) under the filename "artifact.jpg". http://www.histonet.org/site_images_frame.asp Thanks again, Jason _______________________________________________ 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 fjones <@t> namsa.com Tue Nov 9 14:50:25 2004 From: fjones <@t> namsa.com (Fawn Jones) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] H&E procedures Message-ID: <915E55B02E236E4D95258B181EEF6317075D5C@namsams01.namsa.int> Does anybody know of a good H&E procedure for plastics that they are willing to share? We are not staining slides we are staining the blocks themselves. We are told these blocks are a polyester plastic, but we are willing to try any procedure available for any type of plastic such as GMA, MMA, & Technovit 7200 if no one knows of a procedure for polyester plastic. Any help is appreciated, we need to do this stain ASAP, and we are clueless on what we should do. Thank you Fawn Jones From failm <@t> musc.edu Tue Nov 9 15:36:29 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Antibody Help microphthalmia and CD34 Message-ID: We use CD34 from Becton-Dickinson, at 1:100, Hier with Citrate buffer and MITF from Novocastra, 1:10, HIER with Citrate Buffer Rena Fail Medical University of SC Charleston, SC >>> Caldwell 11/09/04 12:58PM >>> Hi Everyone, I am looking for a good antibody for CD34 and microphthalmia-associated transcription factor. We have tried a couple of vendors for the CD34, and the pathologist hasn't been happy with them. Any suggestions would be greatly appreciated. Thank you in advance for your help. Louri Roberts-Caldwell --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From failm <@t> musc.edu Tue Nov 9 15:47:21 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] antigen for B-cells in bone marrow Message-ID: CD20- clone L-26,DAKO, 1:2000 HIER with Citrate Buffer. We do it on BMs almost every day, beautiful every time Rena Fail >>> Sharon Cooperman 11/05/04 06:42PM >>> Dear Histonetters, Does anyone know of an antibody that works well on B-cells in bone marrow? I'm actually trying to use it on a western blot, but I think many Abs that work on IHC work on western. I've tried CD11a, CD11b, and Pax-5, all of which are supposed to work on western but none did (maybe I need to load more protein). Any suggestions would be greatly appreciated. Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From failm <@t> musc.edu Tue Nov 9 15:44:11 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Re: Dumbing down the practical. Message-ID: '77 Peggy, had to do both, Rena Fail >>> 11/06/04 10:15PM >>> For those who remember doing both an H&E and a special stain on the same block - any chance this was BEFORE 1980, when I took my registry exam and started helping students? In my original response to histonet, my note about ASCP not requiring both an H&E and a special stain on the same block was in reference to my time frame (post-1980). I have heard from others, who took their exams in say the 1960's, that did have to do both. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "LuAnn Anderson" To: "Susan Owens" ; "Histonet" Sent: Friday, November 05, 2004 2:52 PM Subject: Re: [Histonet] Re: Dumbing down the practical. > I also remember being required to do specials and H&E's on the same blocks. > If it turned out to be a bad section/sections--or bad tissue--it was back > to the drawing board!! Recuts or new tissue were then in order!! AND, not > to date myself, but everything was done by hand with the exception of > processing!! I too,believe that it is important have that "hands on" know > how for troubleshooting purposes. JMHO. > LuAnn > > > > At 01:12 PM 11/5/04, Susan Owens wrote: > > >1. In all those years, there has never been a time that both the H&E and a > > >special stain has been requested on the same tissue. That would be double > > >penalizing someone if they had a bad tissue. > > > > > > > >Well I don't agree. It's been a few years, but I seem to remember that I did > >special stains and H&E on select blocks when I took the exam(in 1964).. And > >this thing about a 'double penalizing someone' for a bad tissue...Well they > >should not of used 'bad tissue' to begin with. And if they didn't know the > >tissue was 'bad' then they shouldn't of been taking the exam. > >I believe there are too many new faces in histoland that think they know it > >all and in truth they don't have a clue. > > > >ex: 1. A year or two ago I asked someone to make up a % solution of > >something(I don't remember what)..I was told they didn't know how. I asked > >'how could you not know,you have your HT?' They answered "that they didn't > >need to know, where they worked before they purchased everything." > > 2. This year someone I know was taking the practical. According to > >them they were having a problem getting one of the special stains to work. > >SO, after several tries, they just put it on the auto stainer then sent it > >in with all the other slides. > > > >What did this person learn? NOTHING > >And what did ASCP learn? NOTHING (Do they understand they are handing out > >HT's to some people who don't know how to do the work?) > > > >These are just two examples, I could list allot more....But my blood > >pressure would go wild. > > > >Just keep smiling. > >Susan > > > > > > > > > > > > > > > > > > > > > > > > > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@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 Rcartun <@t> harthosp.org Tue Nov 9 15:52:12 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] DiffQuik Question again Message-ID: Some pathologists would argue that if inflammation is not present, there is no reason to look for H. pylori. Also, third party payors may question the policy of doing a histochemical stain for bugs without first examining the H&E-stained slide. 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 >>> "GUTIERREZ, JUAN" 11/09/04 02:56PM >>> Who determines when it's not necessary? If a stain is negative it does not necessarily mean it was not warranted. The stain is done to figure out if the bugs are there or not. The clinician needs to know either way. Because even if it's a tumor, did the bugs have a role in the process? 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: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Tuesday, November 09, 2004 1:38 PM To: Richard Cartun; GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again We do Genta on all gastric bxs and then if they are not necessary we credit the charge. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Tuesday, November 09, 2004 2:17 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again Dear Juan: Do all these patients get billed for the Giemsa histochemical stain? 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 >>> "GUTIERREZ, JUAN" 11/09/04 01:29PM >>> We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@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 ** 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 From Rcartun <@t> harthosp.org Tue Nov 9 15:56:30 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Antibody Help microphthalmia and CD34 Message-ID: A 1:10 dilution of an antibody that probably costs a few hundred dollars doesn't sound very economical. How long are you incubating? 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 >>> "Mildred Fail" 11/09/04 04:36PM >>> We use CD34 from Becton-Dickinson, at 1:100, Hier with Citrate buffer and MITF from Novocastra, 1:10, HIER with Citrate Buffer Rena Fail Medical University of SC Charleston, SC >>> Caldwell 11/09/04 12:58PM >>> Hi Everyone, I am looking for a good antibody for CD34 and microphthalmia-associated transcription factor. We have tried a couple of vendors for the CD34, and the pathologist hasn't been happy with them. Any suggestions would be greatly appreciated. Thank you in advance for your help. Louri Roberts-Caldwell --------------------------------- Do you Yahoo!? Check out the new Yahoo! Front Page. www.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 LuckG <@t> empirehealth.org Tue Nov 9 16:15:20 2004 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Recommendations frozen section Message-ID: Muhammad, I obtained a short VHS format Histology recruiting video from the NSH (www.nsh.org) several years ago. I use it, a 35mm slide show I put together and brochures (the brochures I got from www.ascp.org) when I make presentations to school classes or other interested groups. It's not exactly a "PowerPoint" presentation but still keeps them inviting me back. The intro into the video begins with a simple reenactment of a frozen section process with the specimen from the OR suite to the cryostat to the microscope and final reporting of the results to the surgeon. If it's still available it could be a starting point for you and get your creative juices flowing. Good luck, 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: Muhammad Tahseen [mailto:tahseen@brain.net.pk] Sent: Tuesday, November 09, 2004 1:26 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Recommendations frozen section Dear All We are going to prepare a tutorial about the frozen section for our management staff. Does anybody have any documents or recommendations that they would be willing to share. Thanks in advance Tahseen Laboratory Supervisor SKMCH & RC Lahore, Pakistan. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bills <@t> icpmr.wsahs.nsw.gov.au Tue Nov 9 16:05:50 2004 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Histological dissection of bone In-Reply-To: Message-ID: <000001c4c6ac$12476580$83a7080a@wsahs.nsw.gov.au> RE: [Histonet] Histological dissection of boneJim and Terry, The saw we have is manufactured in Australia by Gemmmasta, 205 Richmond Rd, Richmond, South Australia. Unfortunately this company is no longer around. However, a fellow I spoke to from a lapidary club says that any saw which has the blade lubricated by water is fine and any tool manufacturer can supply these. Similar to brick cutters. All the best Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: Smith, Jim [mailto:Jim.Smith@nuth.northy.nhs.uk] Sent: Wednesday, 10 November 2004 3:04 AM To: 'bills@icpmr.wsahs.nsw.gov.au' Subject: RE: [Histonet] Histological dissection of bone -----Original Message----- From: Coaker, Terry Bill I was interested to read you e-mail to Terry about the use of diamond impregnated "gem" saws; could you possibly let me have some product details, so that I can do some follow-up ? Our greatest need is to have a cutting device that will safely cope with large resections such as the various long bones, sternum, scapula and so on. Cheers Jim Sent: 09 November 2004 09:45 To: Dildey, Petra; Smith, Jim Subject: FW: [Histonet] Histological dissection of bone -----Original Message----- From: Bill Sinai [mailto:bills@icpmr.wsahs.nsw.gov.au] Sent: 08 November 2004 20:49 To: 'Coaker, Terry'; histonet (E-mail) Subject: RE: [Histonet] Histological dissection of bone Terry, For the last 15 years we have been using a "gem" saw just like they use to cut rocks etc prior to polishing. It has a diamond impregated thin blade 1mm thick and 30cm diameter and a clamping mechanism to hold the material (needs some modification for the shape of bone). The specimen is automatically driven towards the blade. No need for fingers anywhere near it. It will cut specimens up to 12cm in height. We replace the blade about every two years. It gives a beautiful clean cut with little heat being generated during the cutting. Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Coaker, Terry Sent: Tuesday, 09 November 2004 2:30 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Histological dissection of bone For many years we have "trimmed" bone samples on a band-saw to obtain slices for decalcification. The equipment needs replacing. A risk assessment indicated that putting a long bone through a band-saw in this way could lead to someone getting hurt! Is there a safer way? Diamond saws are fine for trimming small pieces but what about femur and humerus? How do people handle them safely? Thankyou Terry Terry Coaker Histopathology Operations Manager Cellular Pathology Royal Victoria Infirmary Queen Victoria Road Newcastle upon Tyne NE1 4LP 0191 282 9120 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From c.gorrie <@t> unsw.edu.au Tue Nov 9 16:43:46 2004 From: c.gorrie <@t> unsw.edu.au (Cath Gorrie) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] anti-fluorescein antibody Message-ID: G'Day, I was wondering if anyone has experience with anti-fluorescein antibodies. We are hoping to target cells that were labelled with CDFA-SE some time ago but that are no longer fluorescing. These are in slide adhered frozen tissue sections. I'm interested in dilutions, background, specificity. Any comments would be appreciated. In theory this might work, in practice I have no idea........ Cheers, 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 ---------------------------------------------------------------------------- From Linresearch <@t> aol.com Tue Nov 9 20:42:49 2004 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] LacZ Fixative Message-ID: <36.6653a04e.2ec2da29@aol.com> Hello, Can anyone with experience recommend the best fixative for the B-Gal procedure for the LacZ reporter gene in rodent tissues? Thanks. Lin From JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk Wed Nov 10 03:47:40 2004 From: JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk (JOHN PHILLIPS) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] antigen for B-cells in bone marrow Message-ID: <166A1E642B5B644DA694C08FD29D0ADC603438@ztroy.new-tr.wales.nhs.uk> I note someone has advocated CD20 staining for Bcell in bone marrow: I do not profess to know much about these matters but from my limited experience in performing immuno work on trephine biopsies i think that early Bcell forms may be missed if CD20, is investigated on its own. CD79 antibody will label early B cell precursor cells and so should be performed in conjunction with or perhaps instead of CD20. That is where my knowedge and understanding fails me. I have chosen to contact the "Histonetters" who i am sure will sort this out and as a consequence i will learn from sticking my oar in. That's what this is all about, ain't it. John, Wrexham, Wales. -----Original Message----- From: Mildred Fail [mailto:failm@musc.edu] Sent: 09 November 2004 21:47 To: scoop@mail.nih.gov; histonet@pathology.swmed.edu Subject: Re: [Histonet] antigen for B-cells in bone marrow CD20- clone L-26,DAKO, 1:2000 HIER with Citrate Buffer. We do it on BMs almost every day, beautiful every time Rena Fail >>> Sharon Cooperman 11/05/04 06:42PM >>> Dear Histonetters, Does anyone know of an antibody that works well on B-cells in bone marrow? I'm actually trying to use it on a western blot, but I think many Abs that work on IHC work on western. I've tried CD11a, CD11b, and Pax-5, all of which are supposed to work on western but none did (maybe I need to load more protein). Any suggestions would be greatly appreciated. Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r rheolwr systemau wybod drwy ddefnyddio'r manylion isod. Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain Cymru. Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau???r Ddeddf Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru ar www.newalesnhstrust.org.uk This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the system manager using the details below. The contents of this email represent the views of the individual(s) named above and do not necessarily represent the views of the North East Wales NHS Trust. Please be aware that, under the terms of the Freedom of Information Act 2000, the North East Wales NHS Trust may be required to make public the content of any emails or correspondence received. For futher information on Freedom of Information, please refer to the North East Wales NHS Trust website at www.newalesnhstrust.org.uk For further assistance, please contact system.administrator@new-tr.wales.nhs.uk. From lpwenk <@t> sbcglobal.net Wed Nov 10 03:57:36 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Histotechs on the net website References: <000401c4c693$175a42e0$83020a0a@IHCTech> Message-ID: <007d01c4c70b$b5eea4a0$492cd445@domainnotset.invalid> This is Peggy Wenk. The "Histotech's Home Page" is no longer no line due to the passing of Steven Slap. I helped Steven by finding new sites and checking the links once a year to make certain they still worked and suggesting changes and advising him on the "histology" content of the page. Steven did all the work of typing and maintaining the page. We communicated via email a couple of weeks before his death. He was looking for corporate sponsors to help cover the monthly cost of the site (this had been going on for a couple of years). As of that date, no companies had come forward. I never had access to the site, nor knew who the ISP was, other than some little country (Tonga??? Togo???) where he had vacationed. So much of the material is lost. I have some pages that I had printed out, to check the links. I'm not certain what, if anything, I will do with this material. I've never done a web page and right now don't have the time, with new students. I might think about it after March. I miss Steven, miss our page, and miss the fun we had discovering new sites of interest/help to histotechs. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Patsy Ruegg" To: "'Luis Chiriboga'" ; "'Histonet'" Sent: Tuesday, November 09, 2004 2:34 PM Subject: RE: [Histonet] Histotechs on the net website > I don't know, Peggy Wenk was helping with that I thought??? > Patsy > > -----Original Message----- > From: Luis Chiriboga [mailto:Luis.Chiriboga@med.nyu.edu] > Sent: Tuesday, November 09, 2004 11:12 AM > To: Patsy Ruegg; 'Histonet' > Subject: RE: [Histonet] Histotechs on the net website > > > my apologies to all, for some reason I never made the association, I > guess because I had never met him. perhaps a downside to the internet > world...... Thank you patsy, Will the info still be available through > some other site, it would be even a greater shame to lose that great > resource as well? Luis > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy > Ruegg > Sent: Tuesday, November 09, 2004 12:35 PM > To: 'Luis Chiriboga'; 'Histonet' > Subject: RE: [Histonet] Histotechs on the net website > > > That was Steve Slap who is no longer with us. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Luis > Chiriboga > Sent: Tuesday, November 09, 2004 6:53 AM > To: Histonet > Subject: [Histonet] Histotechs on the net website > > > Morning all, > There used to be a website/page that listed (for those who registered) > histotech's around the country. I believe it was called the > "histotech's homepage" or the "histotech's portal". All the links that > I had are dead and they send you to http://www.histology.to . Does > anyone know what happened to the site? Does anyone know who the > webmaster or administrator was? Thanks in advance L > _______________________________________________ > Histonet mailing list > Histonet@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 lpwenk <@t> sbcglobal.net Wed Nov 10 04:10:54 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] NSH Teleconference plug Message-ID: <009101c4c70d$9142e880$492cd445@domainnotset.invalid> OK - blatant (hopefully mild) selling of the 2005 NSH teleconferences. So click "delete" now if not interested. The 2005 Schedule is available on the web for the NSH Teleconferences. http://www.nsh.org/education/teleconf2002.html For $90 for each teleconference, you can have as many people attend as you wish, each earning 1 CE for each teleconference. (Invite non-histotechs, too, like med techs, cytotechs, lab assistants, pathologists, etc.) Received is a handout for you to duplicate, usually 35-mm slides, plus a sign up sheet and comments sheets. Send in the sign up sheets and comment sheets to NSH (mail or fax) after the meeting. For an extra $10 for each teleconference, you will receive an audio tape about 1 month later, along with a 4 question multiple choice test. For up to 2 years after the teleconference, you can have people listen to the tape, read the handouts, view the slides, and then complete the quiz. Turn in the quiz to NSH, and they earn 1 CE. Good for those who had to stay in the lab working during the teleconference, or those on vacation or medical leave, or the person you hire 4 months from now. Topics for the next couple of months: Dec. 15 - Blood Borne Pathogens - Maureen Doran, BA, HTL(ASCP) Jan 19 - Mohs Surgery for Histotechs - Cliff Chapman, HTL(ASCP)QIHC Feb. 16 - QC, QA, QI - Sandra Dolar, BA, CT(ASCP) Mar. 16 - Grossing for the Histotech - Michael LaFriniere, PA, HT(ASCP) OK - selling job is over. You may now go back to your regularly scheduled Histonet. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 From Julie.Sanders <@t> med.va.gov Wed Nov 10 06:27:12 2004 From: Julie.Sanders <@t> med.va.gov (Julie.Sanders@med.va.gov) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Kappa and Lambda Probes on the Ventana Benchmark Message-ID: <457381D92B01BD44B21CF37CC02EBDFD28E9B7@vhacinexc2.v10.med.va.gov> Is anyone doing Kappa and Lambda ISH on the Ventana Benchmark? If so would it be possible to share the protocols. I have the probes and the product inserts but no hints on a protocol. I was told to skip the signal clarifier, but since it's part of the kit, am not sure how this is done. Any help/info would be greatly appreciated. Julie Julie Sanders, Supervisor, Anatomic Pathology VAMC Cincinnati, Oh. From gu.lang <@t> gmx.at Wed Nov 10 07:45:41 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] ihc-stainer Message-ID: <000a01c4c72b$91fb1950$eeeea8c0@server> In the dako-handbook is a notice about immunstainers with a capacity of 500 slides per run. I wasn't able to find them in the internet. Do you know the name of such a stainer? (just for interest) Gudrun From HornHV <@t> archildrens.org Wed Nov 10 09:07:54 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Re: Dumbing down the practical. Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B990@EMAIL.archildrens.org> I too had to do both in 1973...... 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 Mildred Fail Sent: Tuesday, November 09, 2004 3:44 PM To: ohenry@dfw.net; histonet@lists.utsouthwestern.edu; lpwenk@sbcglobal.net; ander093@tc.umn.edu Subject: Re: [Histonet] Re: Dumbing down the practical. '77 Peggy, had to do both, Rena Fail >>> 11/06/04 10:15PM >>> For those who remember doing both an H&E and a special stain on the same block - any chance this was BEFORE 1980, when I took my registry exam and started helping students? In my original response to histonet, my note about ASCP not requiring both an H&E and a special stain on the same block was in reference to my time frame (post-1980). I have heard from others, who took their exams in say the 1960's, that did have to do both. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "LuAnn Anderson" To: "Susan Owens" ; "Histonet" Sent: Friday, November 05, 2004 2:52 PM Subject: Re: [Histonet] Re: Dumbing down the practical. > I also remember being required to do specials and H&E's on the same blocks. > If it turned out to be a bad section/sections--or bad tissue--it was > back to the drawing board!! Recuts or new tissue were then in order!! > AND, not to date myself, but everything was done by hand with the > exception of processing!! I too,believe that it is important have that > "hands on" know how for troubleshooting purposes. JMHO. LuAnn > > > > At 01:12 PM 11/5/04, Susan Owens wrote: > > >1. In all those years, there has never been a time that both the > > >H&E and a > > >special stain has been requested on the same tissue. That would be double > > >penalizing someone if they had a bad tissue. > > > > > > > >Well I don't agree. It's been a few years, but I seem to remember > >that I did > >special stains and H&E on select blocks when I took the exam(in > >1964).. And > >this thing about a 'double penalizing someone' for a bad > >tissue...Well they > >should not of used 'bad tissue' to begin with. And if they didn't > >know the > >tissue was 'bad' then they shouldn't of been taking the exam. I > >believe there are too many new faces in histoland that think they > >know it > >all and in truth they don't have a clue. > > > >ex: 1. A year or two ago I asked someone to make up a % solution of > >something(I don't remember what)..I was told they didn't know how. I asked > >'how could you not know,you have your HT?' They answered "that they didn't > >need to know, where they worked before they purchased everything." > > 2. This year someone I know was taking the practical. > >According to them they were having a problem getting one of the > >special stains to work. > >SO, after several tries, they just put it on the auto stainer then > >sent it > >in with all the other slides. > > > >What did this person learn? NOTHING > >And what did ASCP learn? NOTHING (Do they understand they are > >handing out > >HT's to some people who don't know how to do the work?) > > > >These are just two examples, I could list allot more....But my blood > >pressure would go wild. > > > >Just keep smiling. > >Susan > > > > > > > > > > > > > > > > > > > > > > > > > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ 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 HornHV <@t> archildrens.org Wed Nov 10 09:10:39 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] DiffQuik Question again Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B991@EMAIL.archildrens.org> There has to be a medical indication for ordering any tests. You can't just do a stain because it is a gastric biopsy. Our hospital does not allow that. We wait on all stains until the pathologist has looked at the H&E. 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 Richard Cartun Sent: Tuesday, November 09, 2004 3:52 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again Some pathologists would argue that if inflammation is not present, there is no reason to look for H. pylori. Also, third party payors may question the policy of doing a histochemical stain for bugs without first examining the H&E-stained slide. 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 >>> "GUTIERREZ, JUAN" 11/09/04 02:56PM >>> Who determines when it's not necessary? If a stain is negative it does not necessarily mean it was not warranted. The stain is done to figure out if the bugs are there or not. The clinician needs to know either way. Because even if it's a tumor, did the bugs have a role in the process? 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: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Tuesday, November 09, 2004 1:38 PM To: Richard Cartun; GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again We do Genta on all gastric bxs and then if they are not necessary we credit the charge. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Tuesday, November 09, 2004 2:17 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again Dear Juan: Do all these patients get billed for the Giemsa histochemical stain? 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 >>> "GUTIERREZ, JUAN" 11/09/04 01:29PM >>> We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@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 ** 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 _______________________________________________ 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 Terry.Marshall <@t> rothgen.nhs.uk Wed Nov 10 09:30:46 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] DiffQuik Question again Message-ID: Really? You don't do specials on kidney or liver until the H&E is examined? 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: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: 10 November 2004 15:11 To: Richard Cartun; juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again There has to be a medical indication for ordering any tests. You can't just do a stain because it is a gastric biopsy. Our hospital does not allow that. We wait on all stains until the pathologist has looked at the H&E. 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 Richard Cartun Sent: Tuesday, November 09, 2004 3:52 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again Some pathologists would argue that if inflammation is not present, there is no reason to look for H. pylori. Also, third party payors may question the policy of doing a histochemical stain for bugs without first examining the H&E-stained slide. 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 >>> "GUTIERREZ, JUAN" 11/09/04 02:56PM >>> Who determines when it's not necessary? If a stain is negative it does not necessarily mean it was not warranted. The stain is done to figure out if the bugs are there or not. The clinician needs to know either way. Because even if it's a tumor, did the bugs have a role in the process? 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: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Tuesday, November 09, 2004 1:38 PM To: Richard Cartun; GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again We do Genta on all gastric bxs and then if they are not necessary we credit the charge. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Tuesday, November 09, 2004 2:17 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again Dear Juan: Do all these patients get billed for the Giemsa histochemical stain? 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 >>> "GUTIERREZ, JUAN" 11/09/04 01:29PM >>> We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@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 ** 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 _______________________________________________ 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 gentras <@t> vetmed.auburn.edu Wed Nov 10 09:35:33 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:17 2005 Subject: Fwd: { SPAM 1 }::[Histonet] Necrotic tissue stain Message-ID: <6.0.1.1.0.20041110093204.01b5f2d0@mailhost.vetmed.auburn.edu> Hello, what about Oncor's, Apop Tag, In Situ Apoptosis Detection Kit (Peroxidase), # S7100-Kit? Best wishes! Atoska >Hello All: > >I have a research investigator who is inquiring about a procedure to >identify necrotic cells. Apparently these cells are not undergoing >apoptosis but rather are appearing swollen, with swollen nuclei and >mitochondria. He believes that they are necrotic but is looking for some >kind of stain or procedure that will distinguish with some specificity >these cells from the normal ones. Any thoughts or help would be greatly >appreciated. > >Susan > > >--------------------------------- From juan.gutierrez <@t> christushealth.org Wed Nov 10 09:49:49 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] DiffQuik Question again Message-ID: We do. Besides, was there not a reason to do the bx in the first place? 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: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Wednesday, November 10, 2004 9:31 AM To: Horn, Hazel V; Richard Cartun; GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again Really? You don't do specials on kidney or liver until the H&E is examined? 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: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: 10 November 2004 15:11 To: Richard Cartun; juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again There has to be a medical indication for ordering any tests. You can't just do a stain because it is a gastric biopsy. Our hospital does not allow that. We wait on all stains until the pathologist has looked at the H&E. 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 Richard Cartun Sent: Tuesday, November 09, 2004 3:52 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again Some pathologists would argue that if inflammation is not present, there is no reason to look for H. pylori. Also, third party payors may question the policy of doing a histochemical stain for bugs without first examining the H&E-stained slide. 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 >>> "GUTIERREZ, JUAN" 11/09/04 02:56PM >>> Who determines when it's not necessary? If a stain is negative it does not necessarily mean it was not warranted. The stain is done to figure out if the bugs are there or not. The clinician needs to know either way. Because even if it's a tumor, did the bugs have a role in the process? 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: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Tuesday, November 09, 2004 1:38 PM To: Richard Cartun; GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again We do Genta on all gastric bxs and then if they are not necessary we credit the charge. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Tuesday, November 09, 2004 2:17 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again Dear Juan: Do all these patients get billed for the Giemsa histochemical stain? 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 >>> "GUTIERREZ, JUAN" 11/09/04 01:29PM >>> We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. 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Thank you. ============================================================================== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bhewlett <@t> cogeco.ca Wed Nov 10 10:02:09 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] antigen for B-cells in bone marrow References: <166A1E642B5B644DA694C08FD29D0ADC603438@ztroy.new-tr.wales.nhs.uk> Message-ID: <001101c4c73e$af269e60$6400a8c0@mainbox> John, You are correct! CD79a exhibits a wider range of B cell staining than CD20, labelling both earlier and later forms. Regards, Bryan ----- Original Message ----- From: "JOHN PHILLIPS" To: "Mildred Fail" ; ; Sent: Wednesday, November 10, 2004 4:47 AM Subject: RE: [Histonet] antigen for B-cells in bone marrow I note someone has advocated CD20 staining for Bcell in bone marrow: I do not profess to know much about these matters but from my limited experience in performing immuno work on trephine biopsies i think that early Bcell forms may be missed if CD20, is investigated on its own. CD79 antibody will label early B cell precursor cells and so should be performed in conjunction with or perhaps instead of CD20. That is where my knowedge and understanding fails me. I have chosen to contact the "Histonetters" who i am sure will sort this out and as a consequence i will learn from sticking my oar in. That's what this is all about, ain't it. John, Wrexham, Wales. -----Original Message----- From: Mildred Fail [mailto:failm@musc.edu] Sent: 09 November 2004 21:47 To: scoop@mail.nih.gov; histonet@pathology.swmed.edu Subject: Re: [Histonet] antigen for B-cells in bone marrow CD20- clone L-26,DAKO, 1:2000 HIER with Citrate Buffer. We do it on BMs almost every day, beautiful every time Rena Fail >>> Sharon Cooperman 11/05/04 06:42PM >>> Dear Histonetters, Does anyone know of an antibody that works well on B-cells in bone marrow? I'm actually trying to use it on a western blot, but I think many Abs that work on IHC work on western. I've tried CD11a, CD11b, and Pax-5, all of which are supposed to work on western but none did (maybe I need to load more protein). Any suggestions would be greatly appreciated. Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mae'r e-bost hwn ac unrhyw ffeiliau a drosglwyddir gydag ef yn gyfrinachol ac wedi'u bwriadu ar gyfer pwy bynnag y cyfeirir ef ato neu atynt. Os ydych wedi ei dderbyn drwy gamgymeriad yna gadewch i'r rheolwr systemau wybod drwy ddefnyddio'r manylion isod. Mae cynnwys yr e-bost hwn yn cynrychioli barn y sawl a enwir uchod, felly nid ydyw'n dilyn ei fod yn cynrychioli barn GIG Gogledd Ddwrain Cymru. Cofiwch fod yn ymwybodol ei bod yn bosibl y bydd disgwyl i Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru roi cyhoeddusrwydd i gynnwys unrhyw ebost neu ohebiaeth a dderbynnir, yn unol ag amodau??Tr Ddeddf Rhyddid Gwybodaeth 2000. I gael mwy o wybodaeth am Ryddid Gwybodaeth, cofiwch gyfeirio at wefan Ymddiriedolaeth GIG Gogledd Ddwyrain Cymru ar www.newalesnhstrust.org.uk This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the system manager using the details below. The contents of this email represent the views of the individual(s) named above and do not necessarily represent the views of the North East Wales NHS Trust. Please be aware that, under the terms of the Freedom of Information Act 2000, the North East Wales NHS Trust may be required to make public the content of any emails or correspondence received. For futher information on Freedom of Information, please refer to the North East Wales NHS Trust website at www.newalesnhstrust.org.uk For further assistance, please contact system.administrator@new-tr.wales.nhs.uk. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From algranth <@t> u.arizona.edu Wed Nov 10 11:25:17 2004 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Oil Red O question Message-ID: <4.3.2.7.2.20041110101921.00c874a8@algranth.inbox.email.arizona.edu> This may be a crazy question. I'm doing mouse livers for a lab here and they requested Oil Red O. Did the stain - looked great. They are asking if it was possible that some of the fat washed away during the washing steps. The experiment requires that they quantitate the fat found in these sections. Is this a possibility? Is there a better way to quantitate fat? For the record - my washing steps are done under a gentle stream of tap water running into a corner of the stain dish or coplin jar - not a full force gush of water from the faucet directly on the slides. Thanks. Andi Grantham ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From tpmorken <@t> labvision.com Wed Nov 10 11:37:12 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Oil Red O question Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA257F6C6@usca0082k08.labvision.apogent.com> Andrea, in short, yes, fat can wash away in this stain. If you want to quantitate, I suggest fixing in formalin and then in osmium BEFORE processing to paraffin ( as you would for EM). Then the fat is immoblized and appears dark grey or greenish on the paraffin H&E section. The morphology is good and quanititation is much easier and more reliable. 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: Andrea Grantham [mailto:algranth@u.arizona.edu] Sent: Wednesday, November 10, 2004 9:25 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Oil Red O question This may be a crazy question. I'm doing mouse livers for a lab here and they requested Oil Red O. Did the stain - looked great. They are asking if it was possible that some of the fat washed away during the washing steps. The experiment requires that they quantitate the fat found in these sections. Is this a possibility? Is there a better way to quantitate fat? For the record - my washing steps are done under a gentle stream of tap water running into a corner of the stain dish or coplin jar - not a full force gush of water from the faucet directly on the slides. Thanks. Andi Grantham ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carrie <@t> ethossearch.com Wed Nov 10 11:40:19 2004 From: carrie <@t> ethossearch.com (Carrie Roy) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Job Opportunity in the Rocky Mountain area Message-ID: Ethos Search Consultants specializes in placing sales, marketing and technical applications candidates, who are scientifically trained, into positions in the life and analytical sciences. We are working on an entry-level field-based technical support position located in the Rocky Mountain area. Our client is a leading supplier of automated diagnostic systems to the anatomical pathology market. Their instrument and reagent systems are used in clinical histology, cytology, and drug discovery laboratories around the world. The Technical Marketing Representative is an entry-level technical support position that is key to the installation and support of company instruments & reagents at customer locations. We are looking for candidates with the following qualifications: ? Associates degree in Applied Sciences or an accredited Medical / Histology Technology course of study with certification. ? 0 ? 2 years histology experience that includes tissue processing, special stains and immunohistochemistry, including work with automated staining instrumentation. ? In-situ hybridization experience is highly desired. ? Training experience is desired but not required. ? Will travel heavily within the region and sometimes throughout the country. If you or anyone you know is interested in this, or any other biotechnology/scientific related position, please feel free to contact me at: Carrie Roy Ethos Search Consultants 4717 Vanderhill Road Torrance, CA 90505 carrie@ethossearch.com 310-791-4396 From Jackie.O'Connor <@t> abbott.com Wed Nov 10 11:53:36 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Oil Red O question Message-ID: I have never done Oil Red O on anything but frozen sections, since the alcohol steps in processing removes the fat. But, it's been awhile - have I missed something new? Jackie "Morken, Tim - Labvision" Sent by: histonet-bounces@lists.utsouthwestern.edu 11/10/2004 11:37 AM To: "'Andrea Grantham'" cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Oil Red O question Andrea, in short, yes, fat can wash away in this stain. If you want to quantitate, I suggest fixing in formalin and then in osmium BEFORE processing to paraffin ( as you would for EM). Then the fat is immoblized and appears dark grey or greenish on the paraffin H&E section. The morphology is good and quanititation is much easier and more reliable. 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: Andrea Grantham [mailto:algranth@u.arizona.edu] Sent: Wednesday, November 10, 2004 9:25 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Oil Red O question This may be a crazy question. I'm doing mouse livers for a lab here and they requested Oil Red O. Did the stain - looked great. They are asking if it was possible that some of the fat washed away during the washing steps. The experiment requires that they quantitate the fat found in these sections. Is this a possibility? Is there a better way to quantitate fat? For the record - my washing steps are done under a gentle stream of tap water running into a corner of the stain dish or coplin jar - not a full force gush of water from the faucet directly on the slides. Thanks. Andi Grantham ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@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 jryan <@t> sleh.com Wed Nov 10 11:56:03 2004 From: jryan <@t> sleh.com (John Ryan) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] I'm sorry I can not reply immediately because I am out of the office, returning on Monday November 2 Message-ID: I'm sorry I can not reply immediately because I am out of the office, returning on Monday November 22, 2004. +++++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 sheriblair1 <@t> netzero.net Wed Nov 10 12:08:22 2004 From: sheriblair1 <@t> netzero.net (sheriblair1@netzero.net) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Gastric Bx staining Message-ID: <20041110.100911.15007.157246@webmail05.lax.untd.com> Where I used to work we would automatically cut a slide for a possible Diff Quick stain. We would wait for the pathologist to give us a list of the cases they wanted stained that day. The unstained slides were discarded after a week. Sheri Blair From tpmorken <@t> labvision.com Wed Nov 10 12:12:31 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Oil Red O question Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA257F6C8@usca0082k08.labvision.apogent.com> Phil, Point taken; you are right that some lipid is lost. What you would have to quantitate in my method is the area of the fat droplets. In my experience, the fat cells retain enough lipid to show the area quite well (we used it to demonstrate fat storage diseases). I suppose real quantitation could be done on Oil-red-O with area and density measurements. However, since the fat moves around a bit during staining and coverslipping, it would not be good for morphometric analysis. If all that is needed is total quantitiation, then non-histology methods would probably be a better choice. Tim Morken -----Original Message----- From: Philip Oshel [mailto:peoshel@wisc.edu] Sent: Wednesday, November 10, 2004 9:59 AM To: Morken, Tim - Labvision Subject: RE: [Histonet] Oil Red O question Tim, I wouldn't trust the quantitation this way. Osmium binds to unsaturated bonds in the lipids, so it doesn't fix saturated lipids all that well -- there's still significant loss of lipids during dehyration. Phil >Andrea, in short, yes, fat can wash away in this stain. If you want to >quantitate, I suggest fixing in formalin and then in osmium BEFORE >processing to paraffin ( as you would for EM). Then the fat is >immoblized and appears dark grey or greenish on the paraffin H&E >section. The morphology is good and quanititation is much easier and >more reliable. > >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: Andrea Grantham [mailto:algranth@u.arizona.edu] >Sent: Wednesday, November 10, 2004 9:25 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Oil Red O question > > >This may be a crazy question. I'm doing mouse livers for a lab here and >they requested Oil Red O. Did the stain - looked great. They are asking >if it was possible that some of the fat washed away during the washing >steps. The experiment requires that they quantitate the fat found in >these sections. Is this a possibility? Is there a better way to >quantitate fat? For the record - my washing steps are done under a >gentle stream of tap water running into a corner of the stain dish or >coplin jar - not a full force gush of water from the faucet directly on >the slides. Thanks. Andi Grantham >..................................................................... >: Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : >: Sr. Research Specialist University of Arizona : >: (office: AHSC 4212) P.O. Box 245044 : >: (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : >: (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : >:...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > >_______________________________________________ >Histonet mailing list >Histonet@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 Marjorie.Lehman <@t> unilever.com Wed Nov 10 12:22:07 2004 From: Marjorie.Lehman <@t> unilever.com (marjorie lehman) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Oil Red O question Message-ID: There are 2 methods, one for frozens and 1 for paraffin, in the 3rd Ed of the AFIP Manual -----Original Message----- From: Morken, Tim - Labvision [SMTP:tpmorken@labvision.com] Sent: Wednesday, November 10, 2004 12:37 PM To: 'Andrea Grantham' Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Oil Red O question Andrea, in short, yes, fat can wash away in this stain. If you want to quantitate, I suggest fixing in formalin and then in osmium BEFORE processing to paraffin ( as you would for EM). Then the fat is immoblized and appears dark grey or greenish on the paraffin H&E section. The morphology is good and quanititation is much easier and more reliable. 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: Andrea Grantham [mailto:algranth@u.arizona.edu] Sent: Wednesday, November 10, 2004 9:25 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Oil Red O question This may be a crazy question. I'm doing mouse livers for a lab here and they requested Oil Red O. Did the stain - looked great. They are asking if it was possible that some of the fat washed away during the washing steps. The experiment requires that they quantitate the fat found in these sections. Is this a possibility? Is there a better way to quantitate fat? For the record - my washing steps are done under a gentle stream of tap water running into a corner of the stain dish or coplin jar - not a full force gush of water from the faucet directly on the slides. Thanks. Andi Grantham ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@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 Marjorie.Lehman <@t> unilever.com Wed Nov 10 12:25:49 2004 From: Marjorie.Lehman <@t> unilever.com (marjorie lehman) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Osmium fat stain Message-ID: The 3rd Ed of the AFIP Manual has a method for paraffin and for frozens. Marge From david.kinsley <@t> spcorp.com Wed Nov 10 12:34:47 2004 From: david.kinsley <@t> spcorp.com (Kinsley, David) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Oil Red O question Message-ID: Hi Andi, I had always performed oil red O on fresh frozen sections. I fixed in 10% NBF after I cut the slides. I did the oil red o stain, counterstained in hematoxylin, rinsed in water and let the slides air dry overnight, then cover slipped with an aqueous mounting medium. Once the aqueous medium is dry, the slides can be placed in xylene and permanently mounted. This way the lipids which dissolve easily in xylene and to some degree in alcohols are intact. If you liver samples are arriving already fixed, you can cryoprotect overnight in 30% sucrose, then snap freeze in isopentane/dry ice. Hope this helps. Dave -----Original Message----- From: Andrea Grantham [mailto:algranth@u.arizona.edu] Sent: Wednesday, November 10, 2004 12:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Oil Red O question This may be a crazy question. I'm doing mouse livers for a lab here and they requested Oil Red O. Did the stain - looked great. They are asking if it was possible that some of the fat washed away during the washing steps. The experiment requires that they quantitate the fat found in these sections. Is this a possibility? Is there a better way to quantitate fat? For the record - my washing steps are done under a gentle stream of tap water running into a corner of the stain dish or coplin jar - not a full force gush of water from the faucet directly on the slides. Thanks. Andi Grantham ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From Lynne.Bell <@t> hitchcock.org Wed Nov 10 12:59:12 2004 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] 100% ethanol shortage Message-ID: Good afternoon all, I have been notified by Pharmco Products that there is a severe shortage of 100% ethanol. The reason has something to do with ethanol being blended into gas as a fuel additive (and probably some other political reasons, as well). My question to my esteemed colleagues is this: do any of you use and combination of ethanol and methanol for 100% and 95% alcohol? One of the sales reps told my supervisor of this concoction. Due to the fact that that the methanol is undrinkable, this "concoction" is not regulated. Do any of you have any information regarding this ethanol shortage or regarding the use of methanol/ethanol in staining and tissue processing. Thanks for your help. Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 From HornHV <@t> archildrens.org Wed Nov 10 13:11:15 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] DiffQuik Question again Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B992@EMAIL.archildrens.org> Yes that is correct. We wait until the H&E's have been looked at. The docs don't always need every stain we used to routinely do. We do go ahead and cut unstained slides so they are ready when we receive orders. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Wednesday, November 10, 2004 9:31 AM To: Horn, Hazel V; Richard Cartun; juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again Really? You don't do specials on kidney or liver until the H&E is examined? 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: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: 10 November 2004 15:11 To: Richard Cartun; juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again There has to be a medical indication for ordering any tests. You can't just do a stain because it is a gastric biopsy. Our hospital does not allow that. We wait on all stains until the pathologist has looked at the H&E. 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 Richard Cartun Sent: Tuesday, November 09, 2004 3:52 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again Some pathologists would argue that if inflammation is not present, there is no reason to look for H. pylori. Also, third party payors may question the policy of doing a histochemical stain for bugs without first examining the H&E-stained slide. 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 >>> "GUTIERREZ, JUAN" 11/09/04 02:56PM >>> Who determines when it's not necessary? If a stain is negative it does not necessarily mean it was not warranted. The stain is done to figure out if the bugs are there or not. The clinician needs to know either way. Because even if it's a tumor, did the bugs have a role in the process? 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: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Tuesday, November 09, 2004 1:38 PM To: Richard Cartun; GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again We do Genta on all gastric bxs and then if they are not necessary we credit the charge. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Tuesday, November 09, 2004 2:17 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again Dear Juan: Do all these patients get billed for the Giemsa histochemical stain? 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 >>> "GUTIERREZ, JUAN" 11/09/04 01:29PM >>> We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@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 ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. 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Saint Josephs Health System, Inc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------ ------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ======================================================================== ====== _______________________________________________ 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 caroline.stott <@t> anatomy.otago.ac.nz Wed Nov 10 13:18:50 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] H&E procedures In-Reply-To: <915E55B02E236E4D95258B181EEF6317075D5C@namsams01.namsa.int > Message-ID: <5.2.1.1.0.20041111081253.020d8ec0@anatomy.otago.ac.nz> Hi Fawn, We use GMA, and this protocol works well. 1. Immerse slides in water 2. Stain in Harris Haematoxylin 30 mins 3. Wash well in running water, with enough time to allow the slides to blue (via the running water) 4. Blot to remove surplus water, and place slides in absolute alcohol 5. Stain in alcoholic eosin (0.1% eosin Y) for 3 mins 6. Complete dehydration (3 more changes absolute alc 1 min each) 7. Clear in xylene and mount with DPX NB if the haematoxylin has stained heavily, leave slides in absolute alcohol at stage 4 fro several minutes to displace some of the excess. Caroline At 15:50 9/11/04 -0500, you wrote: >Does anybody know of a good H&E procedure for plastics that they are >willing to share? We are not staining slides we are staining the blocks >themselves. We are told these blocks are a polyester plastic, but we >are willing to try any procedure available for any type of plastic such >as GMA, MMA, & Technovit 7200 if no one knows of a procedure for >polyester plastic. Any help is appreciated, we need to do this stain >ASAP, and we are clueless on what we should do. >Thank you >Fawn Jones >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Caroline Stott Histology Service Unit University of Otago PO Box 913 Dunedin, New Zealand Ph (03) 479 7152 Fax (03) 479 7136 From haldana <@t> unimoron.edu.ar Wed Nov 10 13:31:40 2004 From: haldana <@t> unimoron.edu.ar (Hernan Aldana Marcos) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] vital stains Message-ID: <002701c4c75b$e70da900$a904a8c0@um.edu> Dear all I am looking for vital stains that could be applied for detection and characterization of lymphatic vesels and ganglia in live animals. I try to undestand the lymphatic systems in armadillos. I like to find betters stains that methilene blue. Thanks in advance Dr. Hern?n J. Aldana Marcos Facultad de Medicina. Universidad de Mor?n Machado 914. B1708JPD. Buenos Aires. Argentina e-mail alternativo hernanjavier@yahoo.com web: http://hjaldanamarcos.bravepages.com http://histologia.bigthicketdirectory.net/main.html From hymclab <@t> hyhc.com Wed Nov 10 14:28:05 2004 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] DiffQuik Question again Message-ID: I agree with Juan. We do Diff-Quiks on all stomach biopsies (not on stomach polyps though). This is not only by the request of our Pathologists, but by the request of the surgeons. In fact they usually write "Gastric biopsy, check for H. pylori" right on the requisition. Dawn -----Original Message----- From: GUTIERREZ, JUAN [mailto:juan.gutierrez@christushealth.org] Sent: Wednesday, November 10, 2004 9:50 AM To: Marshall Terry Dr, Consultant Histopathologist; Horn, Hazel V; Richard Cartun; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again We do. Besides, was there not a reason to do the bx in the first place? 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: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Wednesday, November 10, 2004 9:31 AM To: Horn, Hazel V; Richard Cartun; GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again Really? You don't do specials on kidney or liver until the H&E is examined? 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: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: 10 November 2004 15:11 To: Richard Cartun; juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again There has to be a medical indication for ordering any tests. You can't just do a stain because it is a gastric biopsy. Our hospital does not allow that. We wait on all stains until the pathologist has looked at the H&E. 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 Richard Cartun Sent: Tuesday, November 09, 2004 3:52 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again Some pathologists would argue that if inflammation is not present, there is no reason to look for H. pylori. Also, third party payors may question the policy of doing a histochemical stain for bugs without first examining the H&E-stained slide. 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 >>> "GUTIERREZ, JUAN" 11/09/04 02:56PM >>> Who determines when it's not necessary? If a stain is negative it does not necessarily mean it was not warranted. The stain is done to figure out if the bugs are there or not. The clinician needs to know either way. Because even if it's a tumor, did the bugs have a role in the process? 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: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Tuesday, November 09, 2004 1:38 PM To: Richard Cartun; GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again We do Genta on all gastric bxs and then if they are not necessary we credit the charge. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Tuesday, November 09, 2004 2:17 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again Dear Juan: Do all these patients get billed for the Giemsa histochemical stain? 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 >>> "GUTIERREZ, JUAN" 11/09/04 01:29PM >>> We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@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 ** 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 _______________________________________________ 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Wed Nov 10 15:02:35 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Molecular Biology Lab Message-ID: Greetings fellow 'netters, We are in the process of building our new lab. Does any one have any ideas on building a molecular biology lab? Air requirements, exhaust requirements, etc. Thanks Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX From tahseen <@t> brain.net.pk Wed Nov 10 15:28:40 2004 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] 100% ethanol shortage References: Message-ID: <001801c4c76c$49934480$972bfea9@m7c0y4> Lynne A. Bell, In days of severe shortage of 100% ethanol we used mathanol or denatured alcohol ( 99BP Spirit) in processing and staining. Muhammad Tahseen SKMCH & RC Lahore,Pakistan. ----- Original Message ----- From: Bell, Lynne To: Sent: Wednesday, November 10, 2004 11:59 PM Subject: [Histonet] 100% ethanol shortage > Good afternoon all, > I have been notified by Pharmco Products that there is a severe shortage > of 100% ethanol. The reason has something to do with ethanol being > blended into gas as a fuel additive (and probably some other political > reasons, as well). My question to my esteemed colleagues is this: do > any of you use and combination of ethanol and methanol for 100% and 95% > alcohol? One of the sales reps told my supervisor of this concoction. > Due to the fact that that the methanol is undrinkable, this "concoction" > is not regulated. > > Do any of you have any information regarding this ethanol shortage or > regarding the use of methanol/ethanol in staining and tissue processing. > > Thanks for your help. > > 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 Rcartun <@t> harthosp.org Wed Nov 10 15:38:41 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Re: Polyoma Virus Message-ID: I agree, but the problem with EM in this situation is 1.) polyomavirus infection in the kidney can be extremely focal so it may be missed on EM and 2.) clinicians may need a "same-day" result in order to adjust the patient's immunosuppression. How fast can you turn around your EM? Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> 11/02/04 12:55PM >>> Debra- BK, Polyoma virus, can be demonstrated beautifully with Electron Microscopy. That is, if you have access to an EM lab. I am frequently processing renal core biopsies to r/o BK. If you would like assistance- let me know. Date: Mon, 1 Nov 2004 16:16:43 -0500 From: Browning Deb Subject: [Histonet] polyoma virus, SV40 To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <3AADFB88753AD31189C100902786B91C0E27851F@hch_nt_exchange.hhsc.ca> Content-Type: text/plain; charset="iso-8859-1" Is anyone out there using an antibody against polyoma virus, SV40; BK; or JC, against human tissue for demonstrating kidney rejection? If so, could you share the details, thanks. Debra Browning, ART Technical Specialist, Immunohistochemistry Anatomic Pathology Hamilton Health Sciences phone: (905) 527-4322 ext 46131 e-mail: browning@hhs Theresa R Dobersztyn HT ASCP Electron Microscopy Laboratory Department of Pathology Akron Children's Hospital 1 Perkins Square Akron, Ohio 44308 330-543-8279 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Wed Nov 10 15:39:37 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] 100% ethanol shortage Message-ID: <83AACDB0810528418AA106F9AE9B7F7E5074F1@sjhaexc02.sjha.org> We successfully use Steven's Blends - purchased from Cardinal. There are different brands on the market. Much cheaper than ethanol! Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bell, Lynne Sent: Wednesday, November 10, 2004 1:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] 100% ethanol shortage Good afternoon all, I have been notified by Pharmco Products that there is a severe shortage of 100% ethanol. The reason has something to do with ethanol being blended into gas as a fuel additive (and probably some other political reasons, as well). My question to my esteemed colleagues is this: do any of you use and combination of ethanol and methanol for 100% and 95% alcohol? One of the sales reps told my supervisor of this concoction. Due to the fact that that the methanol is undrinkable, this "concoction" is not regulated. Do any of you have any information regarding this ethanol shortage or regarding the use of methanol/ethanol in staining and tissue processing. Thanks for your help. 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 ** 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 dmccaig <@t> ckha.on.ca Wed Nov 10 15:36:38 2004 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] 100% ethanol shortage Message-ID: <3E5A3F039F0BD8118B4700C00D002024043358@CKHA9> We used to use isopropyl alcohol for the dilute alcohols (75% and 95%) and ethanol for the absolute stations with no problems. -----Original Message----- From: Muhammad Tahseen [SMTP:tahseen@brain.net.pk] Sent: Wednesday, November 10, 2004 4:29 PM To: Bell, Lynne; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] 100% ethanol shortage Lynne A. Bell, In days of severe shortage of 100% ethanol we used mathanol or denatured alcohol ( 99BP Spirit) in processing and staining. Muhammad Tahseen SKMCH & RC Lahore,Pakistan. ----- Original Message ----- From: Bell, Lynne To: Sent: Wednesday, November 10, 2004 11:59 PM Subject: [Histonet] 100% ethanol shortage > Good afternoon all, > I have been notified by Pharmco Products that there is a severe shortage > of 100% ethanol. The reason has something to do with ethanol being > blended into gas as a fuel additive (and probably some other political > reasons, as well). My question to my esteemed colleagues is this: do > any of you use and combination of ethanol and methanol for 100% and 95% > alcohol? One of the sales reps told my supervisor of this concoction. > Due to the fact that that the methanol is undrinkable, this "concoction" > is not regulated. > > Do any of you have any information regarding this ethanol shortage or > regarding the use of methanol/ethanol in staining and tissue processing. > > Thanks for your help. > > 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 From froyer <@t> bitstream.net Wed Nov 10 16:50:38 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] 100% ethanol shortage In-Reply-To: References: Message-ID: <41929B3E.9010208@bitstream.net> HORRORS!! ...say it isn't so! An ethanol shortage so close to the Holidays!! That egg nog is going to be pretty bland! In a pinch, stop by your local liquor store and pick-up a few bottles of "Everclear" (95% ETOH). That's what we use to do to get us through, and there never seemed to be a shortage... ~ Ford Ford Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN Bell, Lynne wrote: >Good afternoon all, >I have been notified by Pharmco Products that there is a severe shortage >of 100% ethanol. The reason has something to do with ethanol being >blended into gas as a fuel additive (and probably some other political >reasons, as well). My question to my esteemed colleagues is this: do >any of you use and combination of ethanol and methanol for 100% and 95% >alcohol? One of the sales reps told my supervisor of this concoction. >Due to the fact that that the methanol is undrinkable, this "concoction" >is not regulated. > >Do any of you have any information regarding this ethanol shortage or >regarding the use of methanol/ethanol in staining and tissue processing. > >Thanks for your help. > >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 slarkham <@t> qltinc.com Wed Nov 10 17:59:32 2004 From: slarkham <@t> qltinc.com (slarkham@qltinc.com) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Re the Oil red O dilemma Message-ID: Hey Andi We cut cryosections and fix in 10% buffered formalin, the sections go into 70% Ethanol quickly and then into the Oil red o, to rinse we go back into 70 for one dip and then into water and mounted with Dako Faramount. Hope that helps Sam From Linresearch <@t> aol.com Wed Nov 10 18:18:22 2004 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] CD Antibodies & Protocols Message-ID: Hi, Would anyone with information on sources for T & B suppressor cells Abs, TNF, IL1, ED1 & ED2 and protocols be willing to share that information? Thanks, Lion From janer <@t> pathology.usyd.edu.au Wed Nov 10 19:12:45 2004 From: janer <@t> pathology.usyd.edu.au (Jane Radford) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Job Opportunity, Australia, Sydney Message-ID: <6.0.1.1.2.20041111115814.01cb57f0@imap.med.usyd.edu.au> There is currently a full time technical officer position available within the Department of Pathology, University of Sydney . The position is with the Tissue Resource Centre a facilitator of neuroscience research. The primary role is to assist with collection and preparation of human brain tissue for tissue banking. Sound knowledge of basic histological sectioning and staining procedures, and a current driver's licence are essential. Please see the following web site for more details of the position: http://bull.ucc.usyd.edu.au/personnel/FMPro Go to "Technical Officer School of Medical Sciences (Pathology)" For any enquires, please contact: Donna Sheedy Research Data Manager NSW Tissue Resource Centre Department of Pathology,D06 University of Sydney,2006 Ph: 61-2-9351 6143 Fax:61-2-9351 3429 donnas@med.usyd.edu.au Jane Radford PhD Histopathology Laboratory Manager Department of Pathology Blackburn Building D06 University of Sydney NSW 2006 Australia Ph: 61 2 9351 6152 Fax: 61 2 9351 3429 From pam <@t> ategra.com Wed Nov 10 20:31:17 2004 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:24:17 2005 Subject: [Histonet] Job Opportunities in Histology Latest Update 11/11/04 Message-ID: Hi Histonetters, Here is the latest update on opportunities with some of my best clients throughout the US who are seeking Histology Supervisors, Histo Technologists and Histo Technicians. These are positions as direct employees of our client. There are fulltime 40 hour per week positions.. As a direct employee of one of our clients you will be provided with full benefits including Health Insurance, Vacation, Sick Pay, Relocation money and a lucrative sign-on bonus. I have part time and temporary positions as well. I have supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are some of my HOTTEST Histology Supervisory positions: 1. Colorado - Histology Manager/PA (I also need a cytology manager/supervisor for this client if you know anyone) 2. Massachusetts - Histology Team Leader 3. California - Immunohistochemistry Supervisor 4. Texas - Pathologists' Assistant 5. NYC, NY - Core Lab Supervisor - Pathology 6. NYC, NY - Pathologists' Assistant Here are some of my HOTTEST Histo Tech positions: 1. Massachusetts - Histo Tech 2. N. California - Histo Tech 3. Texas - Histo Tech 4. SW Florida - Histo Tech 5. South Florida - MOHS Tech 6. South Florida - Histo Tech 7. S. California - Tech Support Histo Tech 8. NYC NY - Histo Tech If you are interested in these jobs, please CALL ME ASAP at 800 466 9919 x234. To speed things up, please also send me a copy of your resume, (if you haven't already done so). If you are interested in jobs outside the above-mentioned areas, please send me your resume as well. I have clients throughout the US. I will keep your resume confidential and will not release it to anyone without your permission (This is Ategra policy as well as my own). My services are at no charge to you. Of course, you may be happy in your present job, but it never hurts to to keep an eye open. Also, if you have friends/peers who might be interested as well, if you could pass my query & name on to them I'd be very grateful. However, if you are interested in any of the jobs above, please call me. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: Pam Barker Senior Lab Recruiter Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd. Winter Park, FL 32792 VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@ategra.com To Learn More About Ategra: http://www.ategra.com -------------------------------------------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. -------------------------------------------------------------------------------------------------- From jkiernan <@t> uwo.ca Thu Nov 11 00:28:01 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Oil Red O question References: <4.3.2.7.2.20041110101921.00c874a8@algranth.inbox.email.arizona.edu> Message-ID: <41930671.3CB773A2@uwo.ca> Ask your client if fat can be washed out by water? You and I know it can't. Persuade your client to buy a textbook and join the Histochemical Society ($50 per year, with a journal arriving every month!) John Kiernan London, Canada. ------------------------------------ Andrea Grantham wrote: > > This may be a crazy question. I'm doing mouse livers for a lab here and > they requested Oil Red O. Did the stain - looked great. They are asking if > it was possible that some of the fat washed away during the washing steps. > The experiment requires that they quantitate the fat found in these > sections. Is this a possibility? Is there a better way to quantitate fat? > For the record - my washing steps are done under a gentle stream of tap > water running into a corner of the stain dish or coplin jar - not a full > force gush of water from the faucet directly on the slides. > Thanks. > Andi Grantham > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lh2 <@t> sanger.ac.uk Thu Nov 11 04:00:44 2004 From: lh2 <@t> sanger.ac.uk (Lindsay Hall) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] NALT fixation, decalcification and staining. Message-ID: <131DF14ECE564A4D8F35376FD3465172043ECCFD@EXCHSRV1.internal.sanger.ac.uk> Hi, I want to look for mouse cell antigens in the NALT from decalcified mouse heads, I know most antibodies to mouse cell surface markers do not work in FFPE tissues. Any suggestions on how to fix the mouse heads and decalcify and still be able to stain for CD4, CD8 etc...? How about Zinc fixation with EDTA decalcification - or will the Zn ions be chelated? Cheers Lindsay From bruyntjes <@t> voeding.tno.nl Thu Nov 11 05:23:32 2004 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] NALT fixation, decalcification and staining. Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079D45@ntexch1.voeding.tno.nl> Linday Ad far as I know it will not be possible to use CD4 on fixed tissues. We have worked some time on NALT too. W have removed the NALT from the nasal cavity and prepared cryosections. Joost Bruijntjes -----Oorspronkelijk bericht----- Van: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Namens Lindsay Hall Verzonden: donderdag 11 november 2004 11:01 Aan: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] NALT fixation, decalcification and staining. Hi, I want to look for mouse cell antigens in the NALT from decalcified mouse heads, I know most antibodies to mouse cell surface markers do not work in FFPE tissues. Any suggestions on how to fix the mouse heads and decalcify and still be able to stain for CD4, CD8 etc...? How about Zinc fixation with EDTA decalcification - or will the Zn ions be chelated? Cheers Lindsay _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From hodges420 <@t> msn.com Thu Nov 11 06:29:05 2004 From: hodges420 <@t> msn.com (MARY T HODGES) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] DiffQuik Question again References: Message-ID: Good morning all, Over the past 7 years, I have seen drs in three states do QS on gastric and liver specials on biospys( only on Hep C or elts for livers). If it not necessary credit to patient account was done before bill and case was signed out. It all had to do with turnaround time for cases. Tere Hodges St Mary's Hospital Tucson, Az ----- Original Message ----- From: Marshall Terry Dr,Consultant Histopathologist To: Horn, Hazel V ; Richard Cartun ; juan.gutierrez@christushealth.org ; histonet@lists.utsouthwestern.edu ; PatPatterson@mhd.com ; JWEEMS@sjha.org Sent: Wednesday, November 10, 2004 7:30 AM Subject: RE: [Histonet] DiffQuik Question again Really? You don't do specials on kidney or liver until the H&E is examined? 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: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: 10 November 2004 15:11 To: Richard Cartun; juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again There has to be a medical indication for ordering any tests. You can't just do a stain because it is a gastric biopsy. Our hospital does not allow that. We wait on all stains until the pathologist has looked at the H&E. 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 Richard Cartun Sent: Tuesday, November 09, 2004 3:52 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again Some pathologists would argue that if inflammation is not present, there is no reason to look for H. pylori. Also, third party payors may question the policy of doing a histochemical stain for bugs without first examining the H&E-stained slide. 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 >>> "GUTIERREZ, JUAN" > 11/09/04 02:56PM >>> Who determines when it's not necessary? If a stain is negative it does not necessarily mean it was not warranted. The stain is done to figure out if the bugs are there or not. The clinician needs to know either way. Because even if it's a tumor, did the bugs have a role in the process? 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: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Tuesday, November 09, 2004 1:38 PM To: Richard Cartun; GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again We do Genta on all gastric bxs and then if they are not necessary we credit the charge. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Tuesday, November 09, 2004 2:17 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again Dear Juan: Do all these patients get billed for the Giemsa histochemical stain? 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 >>> "GUTIERREZ, JUAN" > 11/09/04 01:29PM >>> We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@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 ** 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 _______________________________________________ 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hodges420 <@t> msn.com Thu Nov 11 06:29:48 2004 From: hodges420 <@t> msn.com (MARY T HODGES) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] DiffQuik Question again References: Message-ID: PS they also do the kidneys the same way. Tere Hodges ----- Original Message ----- From: Marshall Terry Dr,Consultant Histopathologist To: Horn, Hazel V ; Richard Cartun ; juan.gutierrez@christushealth.org ; histonet@lists.utsouthwestern.edu ; PatPatterson@mhd.com ; JWEEMS@sjha.org Sent: Wednesday, November 10, 2004 7:30 AM Subject: RE: [Histonet] DiffQuik Question again Really? You don't do specials on kidney or liver until the H&E is examined? 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: Horn, Hazel V [mailto:HornHV@archildrens.org] Sent: 10 November 2004 15:11 To: Richard Cartun; juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again There has to be a medical indication for ordering any tests. You can't just do a stain because it is a gastric biopsy. Our hospital does not allow that. We wait on all stains until the pathologist has looked at the H&E. 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 Richard Cartun Sent: Tuesday, November 09, 2004 3:52 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com; JWEEMS@sjha.org Subject: RE: [Histonet] DiffQuik Question again Some pathologists would argue that if inflammation is not present, there is no reason to look for H. pylori. Also, third party payors may question the policy of doing a histochemical stain for bugs without first examining the H&E-stained slide. 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 >>> "GUTIERREZ, JUAN" > 11/09/04 02:56PM >>> Who determines when it's not necessary? If a stain is negative it does not necessarily mean it was not warranted. The stain is done to figure out if the bugs are there or not. The clinician needs to know either way. Because even if it's a tumor, did the bugs have a role in the process? 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: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Tuesday, November 09, 2004 1:38 PM To: Richard Cartun; GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again We do Genta on all gastric bxs and then if they are not necessary we credit the charge. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Tuesday, November 09, 2004 2:17 PM To: juan.gutierrez@christushealth.org; histonet@lists.utsouthwestern.edu; PatPatterson@mhd.com Subject: RE: [Histonet] DiffQuik Question again Dear Juan: Do all these patients get billed for the Giemsa histochemical stain? 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 >>> "GUTIERREZ, JUAN" > 11/09/04 01:29PM >>> We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@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 ** 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 _______________________________________________ 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mkundu <@t> purdue.edu Thu Nov 11 07:22:45 2004 From: mkundu <@t> purdue.edu (mkundu@purdue.edu) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Sections falling off slides References: <5.2.0.9.2.20041106210425.00bccae8@mail.utexas.edu> Message-ID: <001a01c4c7f1$883b4d10$5296d280@agriculture.purdue.edu> Hi , I am trying to save the thin section of biomaterial from falling off in antigen retival steamer technique. I am using positive charged superfrost slides and I am doing regular microtomy with paraffin blocks . Is there any thing else I can do in addition to prevent from falling? The sections are very small. Any suggestions to improve are welcome. Thank You MK ----- Original Message ----- From: "Brian Dias" To: Sent: Saturday, November 06, 2004 10:13 PM Subject: [Histonet] Sections falling off slides > hi > i'm trying to optimize my immunohistochemistry protocol on slides and am > runnning into a few problems. > 1. i'd like to know if anyone has any strong opinions (either > positive/negative) about conducting such protocols on slides v/s > free-floating. i'd like to be able to do this on slides because mounting > my sections after performing a free-floating technique is virutally > impossible due to the fragile nature and small size of of my sections. > > 2. i run into the problem of my sectiions falling off the slide whilst > doing my ICC > > the brain is dissected form the skull post-perfusion with saline and 4% > paraformaldehyde > it is then stored in 4% paraformaldehyde for 3 days at 4 C > then transferred to 20%sucrose/1X PBS and stored at 4 C for 3 days > before sectioning on the cryostat at a 30-50um thickness and mounted on > Fisher Probe-On Superfrost Plus slides. slides are stored at -80C after > they've dried (approx. 20 mins). > just before (the night before/2 hours before) performing the ICC, i > remove the slides from the freezer and allow to air-dry. > immerse slides in 4% paraformaldehyde for 10 mins at RT in a dish > 1X PBS wash at RT in a dish > then normal ICC steps by overlaying solution on slides and here's where > sections come peeling off. > > i'd appreciate any input on the matter > > thanks a ton > regards > brian > > > > > ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ > ~ ~ ~ ~ ~ ~ ~ ~ > Brian George Dias > > Graduate Student (Crews lab) Office Phone #: 512-475-6738 > The University of Texas at Austin E-mail: > brian.dias@mail.utexas.edu > Institute for Neuroscience Website: > https://webspace.utexas.edu/bgd85/b.htm > Patterson 56 > 1 University Station Stop C0930 > Austin, TX 78712 > ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ > ~ ~ ~ ~ ~ ~ ~ ~ > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ian.montgomery <@t> bio.gla.ac.uk Thu Nov 11 08:10:48 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Golgi-Cox Message-ID: <6.1.2.0.2.20041111140503.02eb3b40@udcf.gla.ac.uk> Camillo Golgi, I know all about him, but who was Cox? In a histology class this morning a student asked me and while I waxed lyrically about old Camillo it was a knuckle dragger as regards Cox. Although smoothly I just replied that he was a co-worker in Golgi's lab. Ian. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk From twebster <@t> nmcinc.org Thu Nov 11 09:58:10 2004 From: twebster <@t> nmcinc.org (Tim Webster) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Free Shandon Varistain 24:4 Rotary Stainer Message-ID: Hi All, We have a Varistain 24:4 going free to the first person who wants it. It is a clean decon'd instrument, that was functioning when removed from service in June 2004. We tried to sell it through one of the 2nd hand equipment companies, but they weren't interested. If nobody takes it, it will be broken up and disposed of. Pay the freight, and it's yours. Give me a call or e-me if you are interested. - Or if you have creative uses for it. - I always thought that in the right environment, it would make a nice planter. :) If nobody calls by Thanksgiving, we will have to offer it as a sacrifice to the Histology gods. Have a great day everybody. Tim Webster Histology Specialist Northwestern Medical Center 133 Fairfield Street St. Albans, VT 05478 (802) 524 1070 twebster@nmcinc.org From cfavara <@t> niaid.nih.gov Thu Nov 11 10:02:13 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Sections falling off slides Message-ID: If you are not drying over night gland do not have time constraints give that a try! 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: mkundu@purdue.edu [mailto:mkundu@purdue.edu] Sent: Thursday, November 11, 2004 6:23 AM To: Brian Dias; Histology Net List Server (E-Mail) Cc: Ramos-Vara, Jose A.; Van Alstine, William G. Subject: Re: [Histonet] Sections falling off slides Hi , I am trying to save the thin section of biomaterial from falling off in antigen retival steamer technique. I am using positive charged superfrost slides and I am doing regular microtomy with paraffin blocks . Is there any thing else I can do in addition to prevent from falling? The sections are very small. Any suggestions to improve are welcome. Thank You MK ----- Original Message ----- From: "Brian Dias" To: Sent: Saturday, November 06, 2004 10:13 PM Subject: [Histonet] Sections falling off slides > hi > i'm trying to optimize my immunohistochemistry protocol on slides and am > runnning into a few problems. > 1. i'd like to know if anyone has any strong opinions (either > positive/negative) about conducting such protocols on slides v/s > free-floating. i'd like to be able to do this on slides because mounting > my sections after performing a free-floating technique is virutally > impossible due to the fragile nature and small size of of my sections. > > 2. i run into the problem of my sectiions falling off the slide whilst > doing my ICC > > the brain is dissected form the skull post-perfusion with saline and 4% > paraformaldehyde > it is then stored in 4% paraformaldehyde for 3 days at 4 C > then transferred to 20%sucrose/1X PBS and stored at 4 C for 3 days > before sectioning on the cryostat at a 30-50um thickness and mounted on > Fisher Probe-On Superfrost Plus slides. slides are stored at -80C after > they've dried (approx. 20 mins). > just before (the night before/2 hours before) performing the ICC, i > remove the slides from the freezer and allow to air-dry. > immerse slides in 4% paraformaldehyde for 10 mins at RT in a dish > 1X PBS wash at RT in a dish > then normal ICC steps by overlaying solution on slides and here's where > sections come peeling off. > > i'd appreciate any input on the matter > > thanks a ton > regards > brian > > > > > ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ > ~ ~ ~ ~ ~ ~ ~ ~ > Brian George Dias > > Graduate Student (Crews lab) Office Phone #: 512-475-6738 > The University of Texas at Austin E-mail: > brian.dias@mail.utexas.edu > Institute for Neuroscience Website: > https://webspace.utexas.edu/bgd85/b .htm > Patterson 56 > 1 University Station Stop C0930 > Austin, TX 78712 > ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ > ~ ~ ~ ~ ~ ~ ~ ~ > > > > > _______________________________________________ > Histonet mailing list > Histonet@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 ajennings <@t> unmc.edu Thu Nov 11 10:19:58 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] LacZ Fixative In-Reply-To: <36.6653a04e.2ec2da29@aol.com> Message-ID: Years of experience with rodent tissues expressing B-Gal. I have had no problems with formalin fixed samples and looking at B-Gal. There was an article in JHC that compared a few fixatives where time was considered one of the most important factors, and glutaraldehyde was the chosen fixative. Journal of Histochemistry and Cytochemistry, Vol. 50, 1421-1424, October 2002, Copyright ? 2002, The Histochemical Society, Inc. Hello, Can anyone with experience recommend the best fixative for the B-Gal procedure for the LacZ reporter gene in rodent tissues? Thanks. Lin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ajennings <@t> unmc.edu Thu Nov 11 10:32:23 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Molecular Biology Lab In-Reply-To: Message-ID: This is no small task Joe. When do you have to turn in your plans? I was an intricate part of the molecular histology lab that was built in the research department at Mayo Clinic in Scottsdale AZ and the Molecular Phenotyping Core at the University of Nebraska Medical Center and would love to share my experiences with you. I am research oriented so if you need JaCHO regulations you may need someone else's input. anita Greetings fellow 'netters, We are in the process of building our new lab. Does any one have any ideas on building a molecular biology lab? Air requirements, exhaust requirements, etc. Thanks Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From twebster <@t> nmcinc.org Thu Nov 11 10:34:53 2004 From: twebster <@t> nmcinc.org (Tim Webster) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Free Varistain has gone. Message-ID: Wow! Who knew! Thanks for the DELUGE of e-mails and calls. We have a winner, as promised the first call back. Thanks to all for your interest. Tim Tim Webster Histology Specialist Northwestern Medical Center 133 Fairfield Street St Albans, VT 05478 802 524 1070 twebster@nmcinc.org From ajennings <@t> unmc.edu Thu Nov 11 10:42:12 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] ready-to-use commercial staining solutions In-Reply-To: <3BC92F29BE821745AB15E04C98EE028D693643@HCH2KMAIL.holy-cross.com> Message-ID: Totally agree with M Rice Poly Scientific not only has quality staining kits, they have great service and sales representation. My volume of special stains is not high, so it saves a lot of tech time having someone else do the QA/QC to make sure that the stained slides I deliver tomorrow are comparable to the ones I did for the same person two years ago. I have had the chance to ask for a special stain that I did not find in the book and was more than pleased that I did not have to carry out the chloroform extraction and the stain worked perfectly. to answer your questions from my lab point of view (research not clinical) The way we purchase our staining solutions, I see no cons to purchasing from a commercial vendor so long as you find one that will work with you on your specific needs. Pros are as stated, save on tech time therefore money saved. The mess and safety issues involved with producing your own staining solutions is alleviated. Not just chemicals but also the glassware and equipment required to produce your own. (I never did find a good way to store a separatory funnel) I use purchased staining solutions but make up the solutions like 3% Acetic Acid etc myself. I keep stains on hand but mostly because I use to make up my own and refuse to throw out powdered stains. I use the control slides from the company initially to set up my own stock of control slides because we use a variety of fixatives and it makes a difference in some of the stains we do. My belief is that it is important for your techs to learn how to make up solutions including chloroform extraction. Not all places they may work will have the option of purchasing their staining solutions and it also gives them an appreciation of what goes into the process. anita Helen, For many years, (more than I care to remember)I prepared all of my own special stain reagents,but as time went on and companies began producing quality reagents I switched over and have never looked back. The company that I have used for many years is Poly Scientific in New York. They can be reached at (631)5860400 and will send you a catalogue. They will also produce custom reagents if you desire. Mike Rice Holy Cross Hospital Ft Lauderdale Florida -----Original Message----- From: Helen Lam [mailto:lam-helen@ctimail3.com] Sent: Monday, November 08, 2004 10:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] ready-to-use commercial staining solutions Dear all, Our lab is thinking about switching to ready-to-use commercial stains. And I think there is no better place than the Histonet to gather opinions form Histo.people, right? 1.. What are the pros and cons of using commercial staining solutions? 2.. Are there anybody out there already using it? Is it possible to replace most if not all of the 'home-made' reagents you regularly use in your stains by commercial products? If yes/no, why? 3.. Even if you are using commercial reagents, is it still necessary to keep a small amount of dyes and chemicals in stock as a 'back-up', just in case the supply of commercial reagents may become uncertain? 4.. How do you test the commerical reagents and prove that it works satisfactorily on your sections / in your applications? 5.. Are the price , quality and shelf-life acceptable? (It would be apprecaited if you could also indicate what brand of what staining solution you are using.) 6.. Considering the price, quality and shelf-life of commerical reagents, do you think they can satisfactorily replace 'home-made' stains in our setting? We have 20-30 slides to do everyday by hand (mainly alcian blue, PAS, a few renal and liver penals every week, some requests for micro-organisms at times). By switching to commercial reagents, we hope that we could reduce the amount or eliminate altogether the need to keep a stock of dangerous chemicals and dye powder. This in turn should save us some storage space, money(especially for keeping stock of rarely used dyes or chemicals that may go bad after years of standing) and man-hours in stain preparation. So what do you think? Any response to any of the above questions are welcome. I look forward to hearing from you. Thanks in advance! Helen Lam Tuen Mun Hospital Hong Kong From akbitting <@t> geisinger.edu Thu Nov 11 11:00:46 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Victoria blue Message-ID: One of our Drs found a lovely photo demonstrating copper binding protein with Victoria blue and asked if this would be a nice alternative to our Orcein. Does anyone use this stain or have a procedure for it? 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 RSRICHMOND <@t> aol.com Thu Nov 11 12:25:34 2004 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Re: DiffQuik Question again Message-ID: <62135811.7ECF2F27.0BF1BB09@aol.com> Several people comment on the appropriateness of doing a stain (such as Diff-Quik) for Helicobacter pylori before the pathologist has seen the H & E slides. In quite a number of pathology services I've worked on in the last few years, it's been customary (assuming the service has any special stain for H. pylori - not all do) to do the stain along with the H & E, on every gastric biopsy specimen submitted. For one thing, manygastroenterologists want the stain done on all their gastric biopsy specimens. And every once in a while, one sees definite Helicobacter in a specimen where no chronic active gastritis (inflammation that includes neutrophils) is to be found. As for liver biopsy specimens (in benign disease), one always needs a trichrome stain to assess fibrosis. I can often do without an iron stain, but once again the clinicians often want it done, for very good reasons - there is more and more concern about hemochromatosis in our patient populations. For billing purposes, it's absolutely necessary that the pathologist's report makes clear that the stain was done and what the results were. Some regulatory agencies also want the control slide results reported. A pathologist can use a canned "boilerplate" text for much of this stain reporting. Bob Richmond Samurai Pathologist From flemons <@t> bhset.org Thu Nov 11 14:19:53 2004 From: flemons <@t> bhset.org (Fran Lemons) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Agitation of cassettes Message-ID: 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 From dbpiontek <@t> clinomicsbio.com Thu Nov 11 14:33:30 2004 From: dbpiontek <@t> clinomicsbio.com (Denise Bland-Piontek) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Pathologists Assistant Opportunity in New York with Biotech Company Message-ID: <145F33B84793CF488578F86355B32E6912FC05@mail.clinomicsbio.com> POSITION: PATHOLOGISTS ASSISTANT SUMMARY: Independently performs gross dissection, description, and histologic sampling of a full range of surgical specimens. Responsible for the supervision and training of histopathology staff in gross room pathology and tissue banking. Will fill the role of Laboratory Manager and work one on one with technical consultant. Will be expected to review slides of normal tissue types and common tumors for quality assurance. Laser Capture Microdissection duties performed as needed. DUTIES AND RESPONSIBILITIES: 1. Performs dissection, description, and histologic sampling of a full range of anatomical specimens. 2. Supervises and trains histopathology staff in gross room pathology methodology and procedures and tissue banking. 3. Performs frozen sections and Laser Capture Microdissection duties as needed, within established protocol and procedural parameters. 4. Coordinates the acquisition of tissue specimens used for research purposes. 5. Maintains a file of image samples using CoolScope, ensuring a high standard of specimen photography. 6. Participates in the acquisition and labeling of specimens, specimen storage and disposal, and the maintenance of reagents and equipment. 7. Performs miscellaneous job-related duties as assigned. MINIMUM JOB REQUIREMENTS: Bachelor's degree with 1 to 3 years experience directly related to the duties and responsibilities specified. Prior knowledge or experience in histopathology laboratory a plus, especially in the areas of antibody characterization and tissue microarray. KNOWLEDGE, SKILLS, AND ABILITIES REQUIRED: * Knowledge of tissue banking, dissection, anatomical description, and histological sampling methods and techniques. * Ability to maintain quality, safety, and/or infection control standards. * Knowledge of gross room pathology procedures, protocols, and techniques. * Records maintenance skills. * Ability to perform frozen sections and Laser Capture Microdissection. * Knowledge of procedures, regulations, and standards for the acquisition of human tissue specimens for research. * Knowledge of accreditation and certification requirements and standards per New York State, CAP and/or CLIA. * Knowledge of quality standards and requirements for pathology specimen photographs utilizing CoolScope. * Ability to supervise, advise, and train staff in area of expertise. * Knowledge of biological specimen disposition, maintenance and storage procedures and techniques. * Knowledge of chemical reagents and equipment used in pathology research. WORKING CONDITIONS AND PHYSICAL EFFORT: * Substantial amount of time will be dedicated to slide review / microscopy. * Moderate physical activity. * Work environment involves exposure to potentially dangerous materials and situations that require following extensive safety precautions and may include the use of protective equipment. Competitive Salary and Benefits *Will have the opportunity to learn state-of-the-art methods, including characterization of novel antibodies, product development and design including tissue array development, photographing routine and IHC slides used in biology database development as well as independent opportunities. Biotech company located on RPI Campus in Watervliet, New York. www.clinomicsbio.com Contact: hr@clinomicsbio.com or fax resume/ CV to (518) 276-3687 Attn: Denise Bland-Piontek / Dr. Patrick Muraca. From sa.drew <@t> hosp.wisc.edu Thu Nov 11 14:42:41 2004 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] CD61 problems Message-ID: I was wondering if anyone else has had any problems with getting CD61(Y2/51) to work on Z-5 fixed bone marrow biopsies. Only 1 out of 8 cases I have tried gave positive staining in the megakaryocytes. The histology lab uses RDO for decal'ing... I believe the process the specimens go through involves the bone marrow lab putting the biopsy in Z-5 for...before transferring them to surgical pathology. The biopsies are then decal'ed for 30 min., rinsed and then put into neutral buffered formalin. Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 From terribraud <@t> msn.com Thu Nov 11 17:07:37 2004 From: terribraud <@t> msn.com (TERRI BRAUD) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] RE: Re: DiffQuik Question again Message-ID: In regards to doing a special stain BEFORE the H&E: A stain is NOT a test. The interrpretation and report of the stained slide/s (and conformation of appropriate stained control/s) by the pathologist is the legal definition of the test by CPT code and is the ONLY basis for billing. Therefore, if a special stain is performed before the H&E is read, it is perfectly acceptable UNLESS the billed portion for either the technical or professional component is issued. In the vast majority of pathology billing, it is not. Even if the bill is generated at stain order, it usually goes to a type of holding file until the case is completed. Even if the stain is ordered, performed and read by the pathologist before OR after the H&E, it must be reported within the context of the report or it still cannot be legally billed. So stain away, all you Histotechs! but don't bill before its reported!!!! Terri Terri L Braud, HT(ASCP) Surgical Pathology Manager University of Virginia Health Systems From Susan.BORTOLOTTO <@t> svhm.org.au Thu Nov 11 19:23:03 2004 From: Susan.BORTOLOTTO <@t> svhm.org.au (BORTOLOTTO Susan (SVHM)) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] mast cell antibody Message-ID: I was hoping someone could help me with information on the BD antibody AA4, that supposedly labels immature mast cells in rat. Does this work in paraffin sections, does it label mature mast cells, is it tissue dependant. Any help would be appreciated. Thanks Sue ---------------------------- Susan Bortolotto, PhD Research Officer Bernard O'Brien Institute of Microsurgery 42 Fitzroy St. Fitzroy, 3056 VIC, AUSTRALIA Ph: +61 3 9288 4018 Fax: +61 3 9416 0926 Email: Susan.BORTOLOTTO@svhm.org.au -------------- 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 Chewy71874 <@t> aol.com Fri Nov 12 02:07:41 2004 From: Chewy71874 <@t> aol.com (Chewy71874@aol.com) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Sections falling off slides Message-ID: Are you putting the positive control tissue on the same slide? When we have tissue that is washing, we have the cutters put the control on a separate slide. We use DAKO silanized slides and most tissue stays on very well, but occasionally tissue will wash. We found that what I call "double dipping" when cutting is a problem (dipping once to put control on slide, and again to put patient tissue on slide). When using silanized or Plus slides, it is important not to touch the surface of the slide. Use only deionized or distilled water in the waterbath, and no additives. Also, if putting the control on the same slide as the unknown, dip only the end of the slide that will contain the tissue, drain it with that side down, then pick up the other tissue by dipping the other end, and drain it with that side down. We also blot our silanized slides after picking up the tissue to remove the water that collects under the tissue, allow to dry, then dry in 60 degree oven for 1 hour. Hope this helps. Ellen Yee, HT Immuno Dept. Central Histology Facility Diagnostic Pathology Medical Group 2420 J Street Sacramento, CA 95816 (916) 689-2885 From Pawel.Pasierbek <@t> imp.univie.ac.at Fri Nov 12 02:18:39 2004 From: Pawel.Pasierbek <@t> imp.univie.ac.at (Pawel,Pasierbek) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] HELP!!! - Autofluorescence in the mouse brain - HELP!!!! Message-ID: <1E6148FED8920445AC79AEB51579E5237C3365@EXCHSRV.imp.univie.ac.at> Dear All, I need you help. One of the users of our microscopy facility is doing immunoassaying on mouse brain slices. After PFA fixation he sees autofluorescence from certain regions of the brain. It looks like rather big grains in the cytoplasm. We see it in the FITC and Cy3 channel! Unfortunately it is also visible in the unfixed preps. I would be very grateful for any hint how to get rid of it. It is important that he still is able to do immunostainigs on the process samples. Thank you very much and have a nice day. Best regards from Vienna, Pawel --------------------------------------------------------------- Pawel PASIERBEK, PhD Institute of Molecular Pathology (IMP) Biooptics Department Dr. Bohr-Gasse 7 A-1030 Vienna AUSTRIA Tel. +43 1 797 30 591 Fax +43 1 798 71 53 e-mail: pawel.pasierbek@imp.univie.ac.at From king_of_karaoke <@t> web.de Fri Nov 12 03:49:08 2004 From: king_of_karaoke <@t> web.de (=?iso-8859-1?Q? Tobias=20M=E4tzig ?=) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] ISH on cryosections of adult mouse heads Message-ID: <931889853@web.de> Hello everyone, I'd like to performe ISH an mouse-head cryosections. My problem is that I have to decalcify the head before I can cut it. Has anyone a good suggestion how to fix and decalcify the adult mouse head without destroying the RNA ? Thanks -Tobias- __________________________________________________________ Mit WEB.DE FreePhone mit hoechster Qualitaet ab 0 Ct./Min. weltweit telefonieren! http://freephone.web.de/?mc=021201 From je22r <@t> udcf.gla.ac.uk Fri Nov 12 04:23:54 2004 From: je22r <@t> udcf.gla.ac.uk (Julia Edgar) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] immunostaining for cytosolic GFP Message-ID: <000e01c4c8a1$b686e480$40e9d182@vet> Dear All GFP is expressed in our cells as a free cytosolic protein i.e. it is not tagged onto another protein. I will be grateful for protocols for the detection of cytosolic GFP by immunocytochemistry. Thanks in anticipation. J Edgar University of Glasgow From BMolinari <@t> heart.thi.tmc.edu Fri Nov 12 07:07:57 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Agitation of cassettes Message-ID: 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 From Susan.Walzer <@t> HCAHealthcare.com Fri Nov 12 08:31:16 2004 From: Susan.Walzer <@t> HCAHealthcare.com (Walzer Susan) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] DiffQuik Question again Message-ID: <492A62ABF11DA240B063BC8278A88A34611087@orlex03.orl.medcity.net> Same here, we do it on all gastrics. -----Original Message----- From: GUTIERREZ, JUAN [mailto:juan.gutierrez@christushealth.org] Sent: Tuesday, November 09, 2004 1:30 PM To: Patterson, Pat; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] DiffQuik Question again We do a Giemsa on all gastric bx. And never get questioned. The pathologist does not have to review the H&E before it's done. We usually turn it in with the H&E. 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: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: Tuesday, November 09, 2004 10:01 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] DiffQuik Question again Hi all - Thanks for response previously for request for DiffQuik stain protocols. I'm trying to get a sense from all about prodecure/policy -- do the clinical sites routinely perform a DiffQuik on all Gastric Bxs? Or do your Pathologists order after initial review of H&E? And if done on all Bxs - have there been any questions from your Billing offices about necessity? Thanks for any information. Pat Patterson Methodist Dallas Medical Center *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Fri Nov 12 09:06:27 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] HELP!!! - Autofluorescence in the mouse brain - HELP!!!! Message-ID: Red Blood cells autofluresce. Sounds like the brains are not perfused to wash out the red blood cells and ensure rapid penetration of fixative. You need to perfuse the whole body at sacrifice to get out all red blood cells. See 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 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 Pawel,Pasierbek Sent: Friday, November 12, 2004 2:19 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HELP!!! - Autofluorescence in the mouse brain - HELP!!!! Dear All, I need you help. One of the users of our microscopy facility is doing immunoassaying on mouse brain slices. After PFA fixation he sees autofluorescence from certain regions of the brain. It looks like rather big grains in the cytoplasm. We see it in the FITC and Cy3 channel! Unfortunately it is also visible in the unfixed preps. I would be very grateful for any hint how to get rid of it. It is important that he still is able to do immunostainigs on the process samples. Thank you very much and have a nice day. Best regards from Vienna, Pawel --------------------------------------------------------------- Pawel PASIERBEK, PhD Institute of Molecular Pathology (IMP) Biooptics Department Dr. Bohr-Gasse 7 A-1030 Vienna AUSTRIA Tel. +43 1 797 30 591 Fax +43 1 798 71 53 e-mail: pawel.pasierbek@imp.univie.ac.at _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jrutherford <@t> sfbr.org Fri Nov 12 09:52:52 2004 From: jrutherford <@t> sfbr.org (Julienne Rutherford) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] restoring dried tissue Message-ID: I work with marmoset monkeys. I have some dried placental tissue - nobody to observe the birth and collect the placenta, so tissue dried out overnight. Is there a way rehydrate dried tissue prior to fixation in formalin, or should I just write it off? I also have some tissue that was collected some years ago that was found in the same opportunistic way, dessicated but placed in formalin as is. Placenta jerky. What can be done with tissue like this? I'm planning to do H & E on my placental samples and am interested in doing IHC for IGF. Thanks! Julienne Rutherford, M.A. Ph.D. candidate Research Assistant Southwest Foundation for Biomedical Research San Antonio, TX From jrutherford <@t> sfbr.org Fri Nov 12 10:02:53 2004 From: jrutherford <@t> sfbr.org (Julienne Rutherford) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] FFPE vs. frozen for IHC Message-ID: Hello histonetters. I have a bank of placental tissue that's been collected over the years and fixed and then stored (long-term) in 10% formalin. I want to make H&E slides of these samples. In addition to basic morphological information, I'm interested in IGF production by the placenta so am considering the best route for IHC on these older tissues. I'm also in a position to collect fresh placentas. I have a series of questions and would appreciate any suggestions: 1. Has anyone worked with placenta, esp. monkey placenta before, specifically looking at IGF system? 2. Given the state of the older tissue (stored in formalin for up to 7 years in some cases) what can I expect from both H&E and FFPE-based IHC? 3. Should I begin snap-freezing the new tissue for IHC or is FFPE a better/more convenient route? I had always been under the impression that IHC had to be done on frozen tissue, but this is apparently not the case. Thank you! Julienne Rutherford Research Assistant (Adjunct) Southwest Foundation for Biomedical Research Southwest National Primate Research Center P.O. Box 760549 San Antonio, Texas 78245-0549 phone (210) 258-9864 fax (210) 258-9883 jrutherford@icarus.sfbr.org Ph.D. candidate Indiana University Bloomington, Indiana jnruther@indiana.edu From Michael.Rice <@t> holy-cross.com Fri Nov 12 10:17:57 2004 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] restoring dried tissue Message-ID: <3BC92F29BE821745AB15E04C98EE028D693659@HCH2KMAIL.holy-cross.com> I have always used Ruffers solution, and it works, there are many vaiants of the formula, but your tissue can be restored. Ruffer worked with Egyptian mummies tissue, so if it was good enough for them it should be fine for your tissue.i am attaching my recipe mike rice pathology holy cross hospital ft lauderdale, fl -----Original Message----- From: Julienne Rutherford [mailto:jrutherford@sfbr.org] Sent: Friday, November 12, 2004 10:53 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] restoring dried tissue I work with marmoset monkeys. I have some dried placental tissue - nobody to observe the birth and collect the placenta, so tissue dried out overnight. Is there a way rehydrate dried tissue prior to fixation in formalin, or should I just write it off? I also have some tissue that was collected some years ago that was found in the same opportunistic way, dessicated but placed in formalin as is. Placenta jerky. What can be done with tissue like this? I'm planning to do H & E on my placental samples and am interested in doing IHC for IGF. Thanks! Julienne Rutherford, M.A. Ph.D. candidate Research Assistant Southwest Foundation for Biomedical Research San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. 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 jkiernan <@t> uwo.ca Fri Nov 12 10:58:33 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Golgi-Cox References: <6.1.2.0.2.20041111140503.02eb3b40@udcf.gla.ac.uk> Message-ID: <4194EBB9.459BE3A4@uwo.ca> W. H. Cox (1891) Arch. mikr. Anat. 37:16. (Ref culled from Carlton's Histol. Tech. 4th edn) Invented a Golgi-type method that didn't use a silver salt (unlike Golgi's). I don't know anything else about him. He's not on file at www.whonamedit.com -- ------------------------------- 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 _______________________________ Ian Montgomery wrote: > > Camillo Golgi, I know all about him, but who was Cox? In a > histology class this morning a student asked me and while I waxed lyrically > about old Camillo it was a knuckle dragger as regards Cox. Although > smoothly I just replied that he was a co-worker in Golgi's lab. > Ian. > > Dr. Ian Montgomery, > Histotechnology, > Graham Kerr Building, > Institute of Biomedical & Life Sciences, > University of Glasgow, > Glasgow, > G12 8QQ. > Tel: 0141 339 8855 > Office: 4652 > Lab: 6644. > Pager: 07625 702883 > e-mail: ian.montgomery@bio.gla.ac.uk > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Nov 12 11:36:16 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Agitation of cassettes In-Reply-To: Message-ID: <002101c4c8de$1cd62e30$83020a0a@IHCTech> 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 From gcallis <@t> montana.edu Fri Nov 12 11:56:13 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Murine NALT and CD markers Message-ID: <6.0.0.22.1.20041112104030.01b14ff0@gemini.msu.montana.edu> Dear Lindsay, You will not be able to fix OR decalcify NALT (nasal associated lymphoid tissue) from a mouse and have successful immunostaining. Two options are remove NALT carefully, and do frozen sections - something we have done many times, OR cryosection nasal turbinates in undecalcified state, using CRYOJANE, from Instrumedics. Murine CD4 and CD8 will never work with FFPE nor decalcification, decalcification does NOT work well after zinc TRIS buffer fixation as it will cause excessive swelling of tissue. Fresh tissue frozen sections are best. Removing NALT is not difficult. I can provide more details on this if you need it - privately. Since NALT is on both sides of nasal passages, you have two that will be there when you dissect these out. We snap freeze inside a very small Tissue Tek plastic mold (10 x 10 mm) with NALT on bottom of mold, and palate side facing up. Palate tissue is mostly connective tissue and tends to curl after removal so a tiny, thin layer of OCT in bottom of mold, then embed NALT in thin OCTlayer to flatten it out, add more OCT and quickly freeze by placing mold into a petri dish floating in liquid nitrogen. DO NOT let liquid nitrogen get into petri dish, and support the dish to keep it from tipping over. You will have a very flat face on block, and with care can obtain 50 or more serial sections of NALT. Air dry frozens overnight at RT. We have great success with murine CD markers, particularly with NALT and improved morphology using 75% acetone/25% absolute ethanol mixture to fix dried FS for 5 min at RT but go directly to buffer after fixation, do not air dry again. You wrote: I want to look for mouse cell antigens in the NALT from decalcified mouse heads, I know most antibodies to mouse cell surface markers do not work in FFPE tissues. Any suggestions on how to fix the mouse heads and decalcify and still be able to stain for CD4, CD8 etc...? How about Zinc fixation with EDTA decalcification - or will the Zn ions be chelated? Lindsay 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 GoodwinD <@t> pahosp.com Fri Nov 12 12:05:22 2004 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Attn: Histology Professionals in NJ and Surrounding Areas Message-ID: <992899E9EC268548AB8DDE246AF88473055F4BD2@PAHEX01.uphs.upenn.edu> Do you live or work in New Jersey or neighboring states? The New Jersey Society for Histotechnology needs you! Annual Membership Dues are only $20 (Students $10). The New Jersey Society for Histotechnology functions to encourage the professional growth and advancement of histotechnologists, and to promote the exchange of knowledge pertinent to the field of histotechnology and related areas. Membership in the NJSH entitles you to receive discounts to society meetings, publications of our newsletter, and the annual membership directory which contains useful information and resources for histotechnologists. To join the New Jersey Society for Histotechnology: Please visit the website at http:// www.njsh.org , click on "Membership", scroll down to the bottom of the page and click on "Apply Now". Fill out the Membership Application form, print it and mail with your check to the following address: New Jersey Society for Histotechnology, P.O. Box 31, Beachwood, New Jersey 08722 Join now and help support our profession in the New Jersey area! From lesley <@t> vancouverbc.net Fri Nov 12 12:27:48 2004 From: lesley <@t> vancouverbc.net (Lesley Weston) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Agitation of cassettes In-Reply-To: Message-ID: When decalcifying in EDTA, I used to keep the cassettes stirring in a beaker of EDTA on a magnetic stirrer in the cold room. It worked pretty well, but I can't cite a reference - that is just the way we do it in that lab. Lesley Weston. on 11/11/2004 12:19 PM, Fran Lemons at flemons@bhset.org wrote: > 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 From making <@t> nersp.nerdc.ufl.edu Fri Nov 12 12:35:37 2004 From: making <@t> nersp.nerdc.ufl.edu (Mike King) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] re: HELP!!! - Autofluorescence in the mouse brain Message-ID: <41950279.2060300@nersp.nerdc.ufl.edu> Pawel, Sounds like you might be seeing lipofuschin granules, common autofluorescent intracellular structures in brain. To reduce autofluroescence see Baschong W, Suetterlin R, Laeng RH. Control of autofluorescence of archival formaldehyde-fixed, paraffin-embedded tissue in confocal laser scanning microscopy (CLSM). J Histochem Cytochem. 2001 Dec;49(12):1565-72. They describe studies using ammonia-ethanol, borohydride, and sudan Black B alone or in combination, with a variety of fixation and section prep methods compatible with immunolabeling. You can find other literature regarding the application of these reagents to autofluorescence problems. Expect some trial-and-error to figure out what works best for your particular specimens. From jkiernan <@t> uwo.ca Fri Nov 12 12:50:06 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] ISH on cryosections of adult mouse heads References: <931889853@web.de> Message-ID: <419505DE.A6EB1BBC@uwo.ca> Yes. Disodium EDTA. It's in all the techniques books more recent than about 1955. John Kiernan London, Canada. ----------------------------------- Tobias M?tzig wrote: > > Hello everyone, > > I'd like to performe ISH an mouse-head cryosections. > My problem is that I have to decalcify the head before I can cut it. > Has anyone a good suggestion how to fix and decalcify the adult mouse head without destroying the RNA ? > > Thanks > > -Tobias- > __________________________________________________________ > Mit WEB.DE FreePhone mit hoechster Qualitaet ab 0 Ct./Min. > weltweit telefonieren! http://freephone.web.de/?mc=021201 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Nov 12 12:58:38 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Reply on HELP!!! - Autofluorescence in the mouse brain In-Reply-To: <41950279.2060300@nersp.nerdc.ufl.edu> References: <41950279.2060300@nersp.nerdc.ufl.edu> Message-ID: <6.0.0.22.1.20041112115646.01b33238@gemini.msu.montana.edu> If you want, I have an excellent review of autofluorescence, the causes and what can or cannot be done to reduce the problem. Although in conjunction with GFP, it is still pertinent to other fluorescent work. I will be happy to attach it if you want, just let me know. At 11:35 AM 11/12/2004, you wrote: >Pawel, >Sounds like you might be seeing lipofuschin granules, common >autofluorescent intracellular structures in brain. To reduce >autofluroescence see >Baschong W, Suetterlin R, Laeng RH. Control of autofluorescence of >archival formaldehyde-fixed, paraffin-embedded tissue in confocal laser >scanning microscopy (CLSM). J Histochem Cytochem. 2001 Dec;49(12):1565-72. >They describe studies using ammonia-ethanol, borohydride, and >sudan Black B alone or in combination, with a variety of fixation and >section prep methods compatible with immunolabeling. You can find other >literature regarding the application of these reagents to autofluorescence >problems. Expect some trial-and-error to figure out what works best for >your particular specimens. > >_______________________________________________ >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 eca9 <@t> georgetown.edu Fri Nov 12 13:00:58 2004 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] fixation with ethanol? Message-ID: <4195086A.5090507@georgetown.edu> Hi, I have a question about fixation. I heard someone taking about using ethanol for fixation? Could someone please provide me with some info on how this can be done? What can it be used for? Just curious.... Eva Georgetown University From emry <@t> u.washington.edu Fri Nov 12 13:19:09 2004 From: emry <@t> u.washington.edu (P. Emry) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] paraffin embedder Message-ID: I would welcome information on new and used paraffin embedders with quotes including shipping to the U of Washington in Seattle. Thank you, Trisha Emry From gcallis <@t> montana.edu Fri Nov 12 13:28:52 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] fixation with ethanol? In-Reply-To: <4195086A.5090507@georgetown.edu> References: <4195086A.5090507@georgetown.edu> Message-ID: <6.0.0.22.1.20041112122138.01b2de80@gemini.msu.montana.edu> Ethanol fixation is can be used for fixation of glycogen, commonly lost in aqueous fixatives. Some use ethanol fixation with some success for fixation of frozen sections for immunohistochemistry. We enjoy that success when it is combined with acetone, but it must be used with a word of caution, ethanol will not work for all immunostaining needs - other fixatives must be tested or optimized for any given antigen. Often it is used in conjunction with other chemicals, such as acetic acid. One famous and popular alcoholic fixative is Carnoys that has ethanol and acetic acid. I suggest you access a good histotechnology textbook - there are a bunch out there and read up on fixation. This fixative is used for larger pieces of tissue destined for paraffin processing. Sheehan and Hrapchak Theory and Practice of Histotechnology although an older text has a excellent discussion on chemicals used for fixation with alcohol included. At 12:00 PM 11/12/2004, you wrote: >Hi, >I have a question about fixation. I heard someone taking about using >ethanol for fixation? Could someone please provide me with some info on >how this can be done? What can it be used for? >Just curious.... >Eva >Georgetown University > >_______________________________________________ >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 liz <@t> premierlab.com Fri Nov 12 17:51:49 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] IFN-y and IL-6 Message-ID: <000101c4c912$935f09d0$76d48a80@AMY> Hello I might need to perform IFN-y and IL-6 immunohistochemistry on decalcifed ankles and knees from a rat model of rheumatoid arthritis. I have searched the literature and have determined that it looks like I can run the IL-6 on decalcifed joint sections. One of the articles used EDTA for decal and the other that I found did not specify the decal agent that was used. Is any one out there familiar with these antibodies and know if they will work in formic acid decalcifed joints? Any help will be appreciated. Thanks in advance 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 jkiernan <@t> uwo.ca Sat Nov 13 00:53:24 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] fixation with ethanol? References: <4195086A.5090507@georgetown.edu> Message-ID: <4195AF64.8EA5E159@uwo.ca> Ethyl alcohol has been used as a fixative from the earliest days of microscopy - before 1700 AD. When microscopes improved (1800s) better fixatives were developed. Many still include ethanol as a major ingredient. Ethanol alone is still useful for fixing smears of cells. There are many books that cover this subject, and Georgetown University has libraries that contain them. Go there and read. -- ------------------------------- 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/ --------------------------------- Eva C Andersson wrote: > > Hi, > I have a question about fixation. I heard someone taking about using > ethanol for fixation? Could someone please provide me with some info on > how this can be done? What can it be used for? > Just curious.... > Eva > Georgetown University > From jnocito <@t> satx.rr.com Sat Nov 13 08:47:32 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Thanks again Molecular Biology Lab References: Message-ID: <018c01c4c98f$b52eb560$286ece44@yourxhtr8hvc4p> Sorry I've been late responding to thank y'all for the information about the molecular biology lab. I have some solid ideas that I can give to the architect and mechanical engineer. Joe the Toe ----- Original Message ----- From: To: "Joe Nocito" Cc: ; "Histonet" Sent: Thursday, November 11, 2004 10:32 AM Subject: Re: [Histonet] Molecular Biology Lab > > > > > This is no small task Joe. When do you have to turn in your plans? I was > an intricate part of the molecular histology lab that was built in the > research department at Mayo Clinic in Scottsdale AZ and the Molecular > Phenotyping Core at the University of Nebraska Medical Center and would > love to share my experiences with you. > I am research oriented so if you need JaCHO regulations you may need > someone else's input. > anita > > > > Greetings fellow 'netters, > We are in the process of building our new lab. Does any one > have any ideas > on building a molecular biology lab? Air requirements, exhaust > requirements, > etc. > Thanks > > > Joe Nocito, BS, HT(ASCP) QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From pruegg <@t> ihctech.net Sat Nov 13 13:20:00 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] 100% ethanol shortage In-Reply-To: <41929B3E.9010208@bitstream.net> Message-ID: I use something called "Reagent Alcohol" which is a mix of ethanol and other (not sure which), from StatLabs and they have not informed me of any shortage, it works fine for all my tissue processing and slide prep needs. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Ford Royer Sent: Wednesday, November 10, 2004 3:51 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] 100% ethanol shortage HORRORS!! ...say it isn't so! An ethanol shortage so close to the Holidays!! That egg nog is going to be pretty bland! In a pinch, stop by your local liquor store and pick-up a few bottles of "Everclear" (95% ETOH). That's what we use to do to get us through, and there never seemed to be a shortage... ~ Ford Ford Royer, MT(ASCP) Midwest Science Biocenter Minneapolis, MN Bell, Lynne wrote: >Good afternoon all, >I have been notified by Pharmco Products that there is a severe shortage >of 100% ethanol. The reason has something to do with ethanol being >blended into gas as a fuel additive (and probably some other political >reasons, as well). My question to my esteemed colleagues is this: do >any of you use and combination of ethanol and methanol for 100% and 95% >alcohol? One of the sales reps told my supervisor of this concoction. >Due to the fact that that the methanol is undrinkable, this "concoction" >is not regulated. > >Do any of you have any information regarding this ethanol shortage or >regarding the use of methanol/ethanol in staining and tissue processing. > >Thanks for your help. > >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 From pruegg <@t> ihctech.net Sat Nov 13 13:30:06 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Sections falling off slides In-Reply-To: <5.2.0.9.2.20041106210425.00bccae8@mail.utexas.edu> Message-ID: Brian if you can cut thinner the sections will stay on slides better, also airdrying over night instead of just 20 minutes would help, if you could fix the sections in something with alcohol in it that might help too, we use a formalin/methanol/acetone mixture, this is a tuff order for sections as thick as you are doing and brain is also hard to keep on the slide. I have a fan that I leave difficult sections on over night to dry, the fan lays flat and slides sit in a rack on their side and the fan blows air on the slides. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Brian Dias Sent: Saturday, November 06, 2004 8:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sections falling off slides hi i'm trying to optimize my immunohistochemistry protocol on slides and am runnning into a few problems. 1. i'd like to know if anyone has any strong opinions (either positive/negative) about conducting such protocols on slides v/s free-floating. i'd like to be able to do this on slides because mounting my sections after performing a free-floating technique is virutally impossible due to the fragile nature and small size of of my sections. 2. i run into the problem of my sectiions falling off the slide whilst doing my ICC the brain is dissected form the skull post-perfusion with saline and 4% paraformaldehyde it is then stored in 4% paraformaldehyde for 3 days at 4 C then transferred to 20%sucrose/1X PBS and stored at 4 C for 3 days before sectioning on the cryostat at a 30-50um thickness and mounted on Fisher Probe-On Superfrost Plus slides. slides are stored at -80C after they've dried (approx. 20 mins). just before (the night before/2 hours before) performing the ICC, i remove the slides from the freezer and allow to air-dry. immerse slides in 4% paraformaldehyde for 10 mins at RT in a dish 1X PBS wash at RT in a dish then normal ICC steps by overlaying solution on slides and here's where sections come peeling off. i'd appreciate any input on the matter thanks a ton regards brian ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ Brian George Dias Graduate Student (Crews lab) Office Phone #: 512-475-6738 The University of Texas at Austin E-mail: brian.dias@mail.utexas.edu Institute for Neuroscience Website: https://webspace.utexas.edu/bgd85/b .htm Patterson 56 1 University Station Stop C0930 Austin, TX 78712 ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Sat Nov 13 13:30:10 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Embedding medium for whole monkey brain sections In-Reply-To: <5DE270EC-305A-11D9-9342-000A95B3CB5E@mail.mcgill.ca> Message-ID: You could use a 6% agar, warm it to get to liquid and then cool it at 4dc to get it to set, you could then freeze the whole sample. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Shahin Zangenehpour Sent: Saturday, November 06, 2004 6:14 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Embedding medium for whole monkey brain sections Dear HistoNetters, I am trying to characterise the expression of FMRP (fragile X mental retardation protein) across an entire monkey brain. The goal of the project is to provide a 3D map of FMRP expression. Due to technical constraints with the Ab-Ag interactions, the best way to carry out the histology is to do free-floating IHC on whole-brain monkey sections (50 microns thick). The obvious problem, of course, is that it is impossible to maintain the relative spatial information in the tissue that is essential for constructing a map out of the histological data because whole sections of the brain often come in more than one piece. The only solution that comes to mind is to embed the entire brain in a matrix that will keep the spatial information intact throughout the histological processing and maintains that information for subsequent analysis. However, I have absolutely no clue as to how I can achieve that. Does anyone know of a method that will allow me to embed the entire brain such that I can freeze and section the brain-matrix blocks, carry out the histology using established free-floating IHC techniques, and then mount the brain-matrix sections on glass slides for subsequent analysis? Many thanks, SZ ________________________________________________________________________ ______________ Shahin Zangenehpour, PhD Postdoctoral Fellow | P. 514 398 6151 | F. 514 398 3255 Psychology Dept | McGill University | 1205 Dr Penfield Ave | Montr?al QC Canada H3A 1B1_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Sat Nov 13 13:40:17 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Zinc. In-Reply-To: <6.1.2.0.2.20041105145141.02ea08b0@udcf.gla.ac.uk> Message-ID: they mean zinc only not zinc formalin unfortunately Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Ian Montgomery Sent: Friday, November 05, 2004 7:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Zinc. When choosing an antibody the companies usually give the preferred method of preparation, frozen, formalin, zinc etc. When they say zinc, do they mean a zinc only fixative or a zinc formalin mixture? Ian. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From aep10 <@t> cornell.edu Sun Nov 14 08:15:18 2004 From: aep10 <@t> cornell.edu (Anna Elisse Beaudin) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] embryo cryosection problem Message-ID: <1333.132.236.104.213.1100441718.squirrel@132.236.104.213> hi all, I recently tried cryosectioning embryos taken at dpc13-17, and had a horrible time because sections were crumbling to pieces, due to what I believe was some really 'crusty' bone or cartiledge sticking out and ripping the tissue as the blade passed over it (this is in the head). I am new to working with embryos, and I am trying to figure out what I did wrong! I fixed embryos 24-48 hours in 4% paraformaldehyde, and then snap froze in OCT. should I have decalcified these specimens? or cryo-protected them? I would greatly appreciate anybody's advice on this! thanks so much in advance! Ana Beaudin Division of Nutritional Sciences Cornell University Ithaca, NY From vanmetmj <@t> pbrc.edu Sun Nov 14 20:10:34 2004 From: vanmetmj <@t> pbrc.edu (Montina Van Meter) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Sections falling off slides Message-ID: Brian, I perfuse rat and mouse brains with 4% paraformaldehyde, post-fix for ~2 hours and then place tissue in 30% sucrose overnight. I routinely section the brains at 40-60 microns and mount them onto chrome-alum subbed slides. I allow the slides to air dry overnight and then place them in the -80 C. freezer. I don't have any problems with the tissue falling off. Using the plus slides should work the same way as long as you let them dry overnight - 20 min. just isn't long enough. (I also wipe away any excess moisture around the sections with kimwipes.) You will not get the same amount of stain penetration as free floating, but it does save on chemicals. Hope this helps... Tina Van Meter Montina J. Van Meter Lab Coordinator Autonomic Neuroscience Pennington Biomedical Research Center Baton Rouge, LA 70808 225-603-0953 vanmetmj@pbrc.edu >>> "Patsy Ruegg" 11/13/04 1:30 PM >>> Brian if you can cut thinner the sections will stay on slides better, also airdrying over night instead of just 20 minutes would help, if you could fix the sections in something with alcohol in it that might help too, we use a formalin/methanol/acetone mixture, this is a tuff order for sections as thick as you are doing and brain is also hard to keep on the slide. I have a fan that I leave difficult sections on over night to dry, the fan lays flat and slides sit in a rack on their side and the fan blows air on the slides. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Brian Dias Sent: Saturday, November 06, 2004 8:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Sections falling off slides hi i'm trying to optimize my immunohistochemistry protocol on slides and am runnning into a few problems. 1. i'd like to know if anyone has any strong opinions (either positive/negative) about conducting such protocols on slides v/s free-floating. i'd like to be able to do this on slides because mounting my sections after performing a free-floating technique is virutally impossible due to the fragile nature and small size of of my sections. 2. i run into the problem of my sectiions falling off the slide whilst doing my ICC the brain is dissected form the skull post-perfusion with saline and 4% paraformaldehyde it is then stored in 4% paraformaldehyde for 3 days at 4 C then transferred to 20%sucrose/1X PBS and stored at 4 C for 3 days before sectioning on the cryostat at a 30-50um thickness and mounted on Fisher Probe-On Superfrost Plus slides. slides are stored at -80C after they've dried (approx. 20 mins). just before (the night before/2 hours before) performing the ICC, i remove the slides from the freezer and allow to air-dry. immerse slides in 4% paraformaldehyde for 10 mins at RT in a dish 1X PBS wash at RT in a dish then normal ICC steps by overlaying solution on slides and here's where sections come peeling off. i'd appreciate any input on the matter thanks a ton regards brian ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ Brian George Dias Graduate Student (Crews lab) Office Phone #: 512-475-6738 The University of Texas at Austin E-mail: brian.dias@mail.utexas.edu Institute for Neuroscience Website: https://webspace.utexas.edu/bgd85/b .htm Patterson 56 1 University Station Stop C0930 Austin, TX 78712 ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ _______________________________________________ Histonet mailing list Histonet@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 John.Auld <@t> whnt.nhs.uk Mon Nov 15 02:50:11 2004 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Vacancies in Melbourne, Australia Message-ID: Dear all (particularly in Oz) A very good young Histotech (British qualified) has moved to Melbourne and is looking for work. Do any susbscribers know of vacancies in the area that I can pass on to him? He trained in this Department and we had high hopes for his future development, but he met a woman and followed her back to Australia, so a department there will get the benefit of him. Many thanks in advance John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral External Tel 0151 604 7025 From jim <@t> hteqa.co.uk Mon Nov 15 03:59:08 2004 From: jim <@t> hteqa.co.uk (Jim Elsam) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Golgi-Cox Message-ID: Hi Ian Harry Cook gives the following reference: Cox WH, (1891) Arch. Mikr.Anat. 37,16 A challenge for your librarian! Golgi published 13 years earlier Best wishes ~~~~~~~~~~~~~~~~~~~~~~ Jim Elsam, HTEQA Services Ltd From nlouisea <@t> gis.net Mon Nov 15 04:30:41 2004 From: nlouisea <@t> gis.net (Nancy Adams) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] non mercury thermometer Message-ID: <001e01c4cafe$29b4b870$e9cb07cf@rutledge> Our lab is eliminating mercury thermometers. The only one we have left is the NBS(National Bureau of Standards) thermometer that is used to calibrate the others. Does anyone know if there is a non mercury NBS thermometer available? Thank you. Nancy Adams From jnocito <@t> satx.rr.com Mon Nov 15 06:30:59 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] non mercury thermometer References: <001e01c4cafe$29b4b870$e9cb07cf@rutledge> Message-ID: <007901c4cb0e$f68b1ff0$286ece44@yourxhtr8hvc4p> Nancy, yes, we purchased ours from VWR. It is black spirit-filled on a yellow background. I'm sorry I don't have the information with now. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Nancy Adams" To: Sent: Monday, November 15, 2004 4:30 AM Subject: [Histonet] non mercury thermometer Our lab is eliminating mercury thermometers. The only one we have left is the NBS(National Bureau of Standards) thermometer that is used to calibrate the others. Does anyone know if there is a non mercury NBS thermometer available? Thank you. Nancy Adams _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From abright <@t> brightinstruments.com Mon Nov 15 09:45:06 2004 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] curling sections Message-ID: I would like to explain a few points that maybe useful to those experiencing problems on the Histonet recently with 'Curling sections' and those who may in the future. There have been quite a number of you mentioning this problem on the Histonet recently, with a range of cryostats from different manufacturers. One of these was manufactured by us, where our user had no problems for years, then the problem started, the outcome was that a new cryostat from another manufacturer was purchased and installed. After 1 month the same problem re-occurred. The sales representative from the company that supplied the new cryostat thought it had to be in the environment (I assume it was not the first time this sales rep had encountered this problem), and the cryostat was moved to a small room with a dehumidifier running constantly. The outcome was that now the sections are fine and not curling. The purpose of this email is to save others from falling into this predicament and saving them from a lot of wasted time/effort and great expense. The cryostat microtome chamber is in fact a dehumidifier, but if routine automatic defrosting is not carried out and the cooling fins become blocked then it becomes inefficient in dehumidifying and it looks like this could cause the above symptoms. I had no answer to this problem when I first looked into it as this was the first reported case of this problem on one of our cryostats in almost 50 years. This was probably due to the highly efficient refrigeration system we install also coupled with our independent specimen temperature control that is supplied to most of our brain sectioning users, allowing them to maintain a much lower chamber, knife and anti-roll plate temperature while controlling the specimen independently at the correct temperature. With the added benefit that the colder chamber temperature acts as an even more efficient dehumidifier. But even with or without these features it is most important to carry out a regular automatic defrost regime, more so in high humidity locations. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Kathleen Spencer [mailto:kspencer@utmem.edu] Sent: 05 November 2004 22:54 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] curling sections Hi Everyone, Several months ago I sent out a cry for help because my perfused frozen 20u sections had started curling up in tight little rolls in the buffer, which made it impossible to do IHC on them. We tried EVERYTHING and finally bought a new cryostat, a Leica 1850. It worked well for one month and then the curling started again. The Leica rep thought it had to be the environment and although it sounded crazy, we went with that. So it was the humidity!!!!!!!!!!!! We have moved the cryostat to a small room with a dehumidifier running constantly. The water catcher resevoir was full after 24 hrs! We were amazed and dumbfounded. So Memphis is the humidity capital of the world I guess. Who would have thought? My sections are nice and flat and beautiful now. I want to thank everyone that tried to help and as I promised, am letting you all know that it was the HUMIDITY! I also put a large beaker of EtOH in the cryostat, and by the way, I love the Leica. It is now in a small room in a different building with a dehumidifier running constantly. Cheers, Kathleen Spencer HT (ASCP) Lab Manager/LCM Supervisor UTHSC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Nov 15 09:53:00 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] IFN-y and IL-6 In-Reply-To: <000101c4c912$935f09d0$76d48a80@AMY> References: <000101c4c912$935f09d0$76d48a80@AMY> Message-ID: <6.0.0.22.1.20041115084852.01b50390@gemini.msu.montana.edu> Liz, You will NOT be able to do IFN gamma on any tissue sections. You need to read Chris van der Loos's publication on this, impossible on tissue sections. As long as IL-6 is membrane bound, you should be ok. Don't even bother with IFN, for that matter IL4 either. I will attach his article to you privately, so you can see why IFN will not work. He gave the NSH International speaker talk on this in Charlotte. If I were to do the IL6, EDTA would be the choice. At 04:51 PM 11/12/2004, you wrote: >Hello > >I might need to perform IFN-y and IL-6 immunohistochemistry on >decalcifed ankles and knees from a rat model of rheumatoid arthritis. I >have searched the literature and have determined that it looks like I >can run the IL-6 on decalcifed joint sections. One of the articles used >EDTA for decal and the other that I found did not specify the decal >agent that was used. Is any one out there familiar with these >antibodies and know if they will work in formic acid decalcifed joints? >Any help will be appreciated. > >Thanks in advance > >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 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Nov 15 10:22:16 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] RE: embryo cryosection problem In-Reply-To: <1333.132.236.104.213.1100441718.squirrel@132.236.104.213> References: <1333.132.236.104.213.1100441718.squirrel@132.236.104.213> Message-ID: <6.0.0.22.1.20041115091732.01b4ce98@gemini.msu.montana.edu> Cryoprotection in 25% - 30% sucrose should improve your sectioning. We make ours up in Dulbeccos PBS, and let the fixed embryos sit in these overnight at 4C, but yours are older at dpc 13 - 17, they could sit longer. Depending on what you are doing (you did not say) decalcification with EDTA is an alternative but you must still cryoprotect after decalcification if you still see minute boney parts ripping out. You could use a tungsten carbide c profile blade to help deal with these minute fragments IF you didn't want to decalcify. Good luck At 07:15 AM 11/14/2004, you wrote: >hi all, > > I recently tried cryosectioning embryos taken at dpc13-17, and had a >horrible time because sections were crumbling to pieces, due to what I >believe was some really 'crusty' bone or cartiledge sticking out and >ripping the tissue as the blade passed over it (this is in the head). >I am new to working with embryos, and I am trying to figure out what I >did wrong! I fixed embryos 24-48 hours in 4% paraformaldehyde, and >then snap froze in OCT. should I have decalcified these specimens? or >cryo-protected them? I would greatly appreciate anybody's advice on >this! thanks so much in advance! > >Ana Beaudin >Division of Nutritional Sciences >Cornell University >Ithaca, NY > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Nov 15 11:21:00 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] SafeClear and SafeClear II xylene substitutes Message-ID: <6.0.0.22.1.20041115101708.01b16db8@gemini.msu.montana.edu> Dear All, I am curious to know if labs are liking these two xylene substitute clearing agents - SafeClear and SafeClear II from Fisher? Have you compared them to Clearite 3 or Propar? If so, did they compare favorably? What do the MSDS call these substitutes? i.e. is there any chemical information included on MSDS for these to indicate they are single aliphatic hydrocarbons, comparable to Clearite 3? Also do you use them for staining? For processing? Thanks in advance 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 TMcNemar <@t> lmhealth.org Mon Nov 15 11:51:10 2004 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Formalin spill cleanup... Message-ID: <90092A4ED388D7119575006008F7112049CBFB@NT_EXCHANGE> Does anyone know of a product for formalin cleanup that also neutralizes in one step? We have one product that solidifies but does not neutralize. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 From David.Edmondson <@t> christie-tr.nwest.nhs.uk Mon Nov 15 12:00:16 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Frozens Message-ID: Someone asked about Frozens last week and I came accross an obituary for Arnold Levene from a copy of The Daily Telegraph in 1998. So I have sent it to histonet.org and its called "Levene". There are a fair few words of interest. Not just a column inch. My apologies that I have mislaid the original message, and can not remember who to thank for giving me the copy in the first place either. Dave Edmondson Christie Hospital Manchester UK From RBARNHART <@t> summithealth.org Mon Nov 15 12:13:29 2004 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Formalin spill cleanup... Message-ID: We use FanPad from American Master Tech (www.americanhistology.com) They come in three sizes #CWFAN55 (8" x 11"), CWFAN614 (6" x14") and CWFAN825 (8" x 25") Becky >>> Tom McNemar 11/15/2004 12:51:10 PM >>> Does anyone know of a product for formalin cleanup that also neutralizes in one step? We have one product that solidifies but does not neutralize. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dobbin <@t> upei.ca Mon Nov 15 12:37:47 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Hepatocytes and CD79a Message-ID: <4198CD4A.28469.490568@localhost> Hello All, Please help a colleague out by answering the following: Are normal hepatocytes positive for CD79a? Cheers! Greg ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Research Technologist, Pathology Lab, Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From la.sebree <@t> hosp.wisc.edu Mon Nov 15 13:57:01 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Hepatocytes and CD79a Message-ID: I wouldn't think so Greg. I'll bet what you're seeing is endogenous biotin; liver is loaded with it and even blocking steps aren't always adequate. 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 Greg Dobbin Sent: Monday, November 15, 2004 9:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Hepatocytes and CD79a Hello All, Please help a colleague out by answering the following: Are normal hepatocytes positive for CD79a? Cheers! Greg ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Research Technologist, Pathology Lab, Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gentras <@t> vetmed.auburn.edu Mon Nov 15 15:07:20 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:18 2005 Subject: Fwd: Re: { SPAM 1 }::[Histonet] non mercury thermometer Message-ID: <6.0.1.1.0.20041115150719.01ba4958@mailhost.vetmed.auburn.edu> >Date: Mon, 15 Nov 2004 15:06:59 -0600 >To: "Nancy Adams" >From: "Atoska S. Gentry" >Subject: Re: { SPAM 1 }::[Histonet] non mercury thermometer > > >Hello, I'm not familiar with NBS thermometers, but my mercury free >thermometers are Fisher Scientific # 15-041-4D. Best wishes! Atoska > > >At 04:30 AM 11/15/2004, you wrote: >>Our lab is eliminating mercury thermometers. The only one we have left >>is the NBS(National Bureau of Standards) thermometer that is used to >>calibrate the others. Does anyone know if there is a non mercury NBS >>thermometer available? >> >>Thank you. >>Nancy Adams >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mark.bliek <@t> bolton-works.com Mon Nov 15 20:52:29 2004 From: mark.bliek <@t> bolton-works.com (Mark Bliek) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Info needed on LKB Historange 2218 and LKB Macrotome Message-ID: <01C4CB5D.67F45780.mark.bliek@bolton-works.com> Dear List, Last week I became the proud owner of 2 decommissioned LKB microtomes. One is a LKB Historange 2218, the other a LKB Macrotome (no misspelling). The 2218 is an automatic, rotating microtome and seems to work fine, but I need a knifeholder. The LKB Macrotome is a computer controlled sliding Microtome. The LKB Macrotome is a beast: 3 man were needed to lift it. The way it has been designed makes me believe that it must be a very accurate microtome, which, despite its 20+ years, I want to use because of it heavy duty construction. The controller is however missing. LKB has disappeared form the market, and a search on the Internet did not yield any useful information on either microtomes. If anyone has information, parts, manuals, technical drawings of these machines, I would be grateful to learn about it. Mark Bliek Bolton Works LLC Phone (860) 646-3948 From bruyntjes <@t> voeding.tno.nl Tue Nov 16 03:44:18 2004 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] (no subject) Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079D4D@ntexch1.voeding.tno.nl> Hi all We have done immunocytochemistry on some slides. An antigen-retrieval (citrate pH 6.0) was needed. But it would be nice if we could use the same slides for another antigen (AR-EDTA pH 10 is recommended). Has anyone of you done this before, and is it possible? Joost Bruijntjes Zeist The Netherlands This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From ccrowder <@t> mail.vetmed.lsu.edu Tue Nov 16 09:05:41 2004 From: ccrowder <@t> mail.vetmed.lsu.edu (Cheryl Crowder) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Neutrophil staining Message-ID: Hello - I have a researcher who wants to stain neutrophils in horse tissue. I have found a fluorescent method but am looking for others. Do any of you know of a stain for neutrophils, either special stain or IHC? Supposedly there is an antibody but I have yet to find it. Thank you in advance for your help. Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA 70803 225-578-9734 FAX: 225-578-9720 From cfavara <@t> niaid.nih.gov Tue Nov 16 09:09:44 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] (no subject) Message-ID: I have done some of this and will share what I have learned. Some questions to ask yourself are: Do you want to maintain the staining already present? Can the second antibody be optimized for the AR that has already been done? Do you know the location of the targets? Different cellular component, cells, infectious agents, etc? Heat will break the bonds that you have for the first reaction and unless the reporter is stable - DAB or ALK PHOS RED in my experience- you will lose any signal that you have currently demonstrated. In my experience a well fixed and dried specimen can withstand 2-3 AR with heat. Sometimes it is easier to just recut and stain separately. Depends on what you are trying to see. Great resource is Chris Van der Loos' book on multiple staining. Good Luck, 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: Bruijntjes, J.P. [mailto:bruyntjes@voeding.tno.nl] Sent: Tuesday, November 16, 2004 2:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Hi all We have done immunocytochemistry on some slides. An antigen-retrieval (citrate pH 6.0) was needed. But it would be nice if we could use the same slides for another antigen (AR-EDTA pH 10 is recommended). Has anyone of you done this before, and is it possible? Joost Bruijntjes Zeist The Netherlands This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Tue Nov 16 09:49:12 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Tbx2 IHC Message-ID: If anyone in the whole wide world has any experience doing IHC with a Tbx2 antibody - I'd love to hear from you. I'm going to be staining human breast, but I need to know more before I proceed. I haven't found any references to Tbx2 staining. Anybody? Jackie O'Connor From pzeitlow <@t> bbpllab.com Tue Nov 16 10:01:56 2004 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] MUC6 antibody Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A0BEAE9@bbplsrv1.bbpl> Does anyone know a laboratory doing this stain on ffpe tissues? Pat Z Department of Molecular Pathology Boyce and Bynum Pathology Labs, P.C. From Jenny-Oblander <@t> omrf.ouhsc.edu Tue Nov 16 10:49:11 2004 From: Jenny-Oblander <@t> omrf.ouhsc.edu (Jenny Oblander) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Creatine Kinase MB Message-ID: Hi All, I am trying to find an antibody to detect CKMB in mouse tissue. Has anyone done this before? Thanks Jenny J.Oblander, HT (A.S.C.P.) Comparative Medicine Oklahoma Medical Research Foundation MS#32 825 NE 13th St. Okc,Ok 73104 jenny-oblander@omrf.ouhsc.edu 405-271-7083 From pruegg <@t> ihctech.net Tue Nov 16 11:06:46 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Neutrophil staining In-Reply-To: Message-ID: <000201c4cbfe$a7a051d0$83020a0a@IHCTech> Cheryl, A good hemato stain like Giemsa would surely show the granules in neutrophils also myleoperoxidase will label the endogenous peroxidase in myleoid cells. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cheryl Crowder Sent: Tuesday, November 16, 2004 8:06 AM To: Histonet Subject: [Histonet] Neutrophil staining Hello - I have a researcher who wants to stain neutrophils in horse tissue. I have found a fluorescent method but am looking for others. Do any of you know of a stain for neutrophils, either special stain or IHC? Supposedly there is an antibody but I have yet to find it. Thank you in advance for your help. Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA 70803 225-578-9734 FAX: 225-578-9720 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Nov 16 11:43:03 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] MUC6 antibody In-Reply-To: <813FB33DA405334F947F8BFC6EBD0B2A0BEAE9@bbplsrv1.bbpl> References: <813FB33DA405334F947F8BFC6EBD0B2A0BEAE9@bbplsrv1.bbpl> Message-ID: <6.0.0.22.1.20041116104023.01b1e200@gemini.msu.montana.edu> Contact Dr. Chris van der Loos at "C.M. vander Loos" - he has done another MUC antibody, and should have some hints on doing this. He did MUC5 (if I remember correctly) as double stain with another antibody. At 09:01 AM 11/16/2004, you wrote: >Does anyone know a laboratory doing this stain on ffpe tissues? > >Pat Z >Department of Molecular Pathology >Boyce and Bynum Pathology Labs, P.C. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Tue Nov 16 11:59:08 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Old style embedding rings, plastic Message-ID: <6.0.0.22.1.20041116105614.01b25560@gemini.msu.montana.edu> I have two boxes of old style white plastic embedding rings for whoever wants them, a freebie. First come, first serve, and please provide a US postal, slowmail address, as these will NOT be shipped via UPS or any other shipping service. Sorry, but these will only be mailed to a laboratory inside the USA, no foreign addresses. 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 Nov 16 12:44:11 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] plastic embedding rings are gone Message-ID: <6.0.0.22.1.20041116114324.01b284e0@gemini.msu.montana.edu> Dear All, Plastic embedding rings are gone 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 TJJ <@t> Stowers-Institute.org Tue Nov 16 13:12:44 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Neutrophil staining Message-ID: Cheryl, CD 15 stains neutrophils, I think. Also using ASD Chloroacetate Esterase-aka Leder stain (Sigma kit) will stain them as well. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From cchiu <@t> itsa.ucsf.edu Tue Nov 16 13:20:40 2004 From: cchiu <@t> itsa.ucsf.edu (Charles Chiu) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Can you use RNAlater or RNAlaterice to extract RNA from OCT-embedded tissues? Message-ID: <002501c4cc11$5c406190$6a0de6a9@LINUS> Hello, I am currently working with OCT-embedded tissues and am wondering if anyone has had success extracting RNA from OCT-embedded tissues or has a protocol for doing so. I am considering RNAlater-ICE by Ambion but specifically -- (1) is OCT fully water soluble and not infiltrative? (2) Should I use RNAlater-ICE as a preservative upon thawing the OCT-embedded tissues or just proceed with TRIzol extraction? (3) Do I need to get rid of the OCT embedding medium and if so, how do I do this? Any help would be greatly appreciated! Thanks, Charles ------------------------------------------------------------------- Dr. Charles Chiu, M.D./Ph.D. Clinical Fellow, Infectious Diseases UCSF School of Medicine 521 Parnassus Ave., Room C443, Box 654 San Francisco, CA 94143-0654 Cell: (415) 420-4463 Pager: (415) 719-0088 cchiu@itsa.ucsf.edu ------------------------------------------------------------------- From MinHan.Tan <@t> vai.org Tue Nov 16 13:41:54 2004 From: MinHan.Tan <@t> vai.org (Tan, MinHan) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Can you use RNAlater or RNAlaterice to extract RNA fromOCT-embedded tissues? Message-ID: Hello, I approached the company for an RNA protocol one year ago; the response was that this 'application' wasn't supported. However, I have had no problems with extraction of RNA from OCT blocks. One can shave off multiple thick sections and then homogenize these sections in Trizol. Alternatively, I usually trim the block by cutting off large pieces of OCT without tissue with a razor. I subsequently homogenize the remaining chunk of tissue mixed with OCT in Trizol. Works great for Affymetrix microarray analysis, and there's generally no need to slice and dice in a cryotome. :) Guess you'll just have to check to make sure that it works for your application. RNA Later-ICE has worked very badly for me. The majority of tissue samples I thawed in this product yielded degraded RNA, though I admit that it was possible that the tissue was degraded prior to that. We had to use it as we wanted to stick a needle into the tissue at that time. So just proceed with Trizol. Min-Han -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles Chiu Sent: Tuesday, November 16, 2004 2:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Can you use RNAlater or RNAlaterice to extract RNA fromOCT-embedded tissues? Hello, I am currently working with OCT-embedded tissues and am wondering if anyone has had success extracting RNA from OCT-embedded tissues or has a protocol for doing so. I am considering RNAlater-ICE by Ambion but specifically -- (1) is OCT fully water soluble and not infiltrative? (2) Should I use RNAlater-ICE as a preservative upon thawing the OCT-embedded tissues or just proceed with TRIzol extraction? (3) Do I need to get rid of the OCT embedding medium and if so, how do I do this? Any help would be greatly appreciated! Thanks, Charles ------------------------------------------------------------------- Dr. Charles Chiu, M.D./Ph.D. Clinical Fellow, Infectious Diseases UCSF School of Medicine 521 Parnassus Ave., Room C443, Box 654 San Francisco, CA 94143-0654 Cell: (415) 420-4463 Pager: (415) 719-0088 cchiu@itsa.ucsf.edu ------------------------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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. From vazquezr <@t> ohsu.edu Tue Nov 16 14:42:40 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Position needed Message-ID: Would anyone know of a Mohs/histotech be interested in a job in San Jose, CA Very new. Would have to set lab up. Not sure pay. Or if willing to go there and train someone to do Mohs. Doctor is a great guy to work for. I will have more info later if anyone is interested. Robyn Vazquez OHSU Portland, Or From Jenny-Oblander <@t> omrf.ouhsc.edu Tue Nov 16 15:16:59 2004 From: Jenny-Oblander <@t> omrf.ouhsc.edu (Jenny Oblander) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] supravital stains Message-ID: Hi again, Does anyone have a procedure for Rhodanile blue.I'm looking for Heinz bodies. Thanks Jenny J.Oblander, HT (A.S.C.P.) Comparative Medicine Oklahoma Medical Research Foundation MS#32 825 NE 13th St. Okc,Ok 73104 jenny-oblander@omrf.ouhsc.edu 405-271-7083 From Linresearch <@t> aol.com Tue Nov 16 16:13:30 2004 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] FFPE In Frog Tissue: Message-ID: <1f3.2701b33.2ecbd58a@aol.com> Hello everyone: Does anyone know of an antibody that will work in FFPE frog tissue for IRIDO Virus. Please respond. Thanks in advance, Lin From t-kuzniar <@t> md.northwestern.edu Tue Nov 16 16:15:23 2004 From: t-kuzniar <@t> md.northwestern.edu (t-kuzniar@md.northwestern.edu) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Luciferase antibody Message-ID: <200411162215.iAGMFiw9007616@casbah.it.northwestern.edu> Hi everyone, I am looking for a nice anti-luciferase antibody for immunohistochemistry for formalin-fixed, paraffin-embedded sections of the lung. Tom Kuzniar t-kuzniar@md.northwestern.edu From gcallis <@t> montana.edu Tue Nov 16 16:54:38 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Luciferase antibody In-Reply-To: <200411162215.iAGMFiw9007616@casbah.it.northwestern.edu> References: <200411162215.iAGMFiw9007616@casbah.it.northwestern.edu> Message-ID: <6.0.0.22.1.20041116155354.01b089f0@gemini.msu.montana.edu> You might want to try Molecular Probes, or contact their tech services for information. At 03:15 PM 11/16/2004, you wrote: >Hi everyone, > >I am looking for a nice anti-luciferase antibody for immunohistochemistry >for formalin-fixed, >paraffin-embedded sections of the lung. > >Tom Kuzniar >t-kuzniar@md.northwestern.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 liz <@t> premierlab.com Tue Nov 16 17:03:44 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Luciferase antibody In-Reply-To: <200411162215.iAGMFiw9007616@casbah.it.northwestern.edu> Message-ID: <000001c4cc30$85e720f0$76d48a80@AMY> Tom I did some work in the past with RDI's goat-anti-Luciferase. But from what I remember it was a bit difficult to work with and I was not that pleased with the results. I believe I had some issues with non-specific staining. I do have a protocol that you can start with and some images if you would like them. The sections I used were paraformaldehyde fixed paraffin embedded mouse lung, kidney and liver. Good Luck Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of t-kuzniar@md.northwestern.edu Sent: Tuesday, November 16, 2004 3:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Luciferase antibody Hi everyone, I am looking for a nice anti-luciferase antibody for immunohistochemistry for formalin-fixed, paraffin-embedded sections of the lung. Tom Kuzniar t-kuzniar@md.northwestern.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From conlonb <@t> comcast.net Tue Nov 16 17:41:04 2004 From: conlonb <@t> comcast.net (Brian D. Conlon) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] coverslipping with tape Message-ID: <001601c4cc35$bf41a5a0$6401a8c0@gateway> hi everyone,does anyone know how to save slides that have been coverslipped with tape on a sakura coverslipper? we pulled some slides that had been filed for a couple of years and the tape is loose and the sections have adhered to the tape instead of the slide. any suggestions? thank you in advance. From jkiernan <@t> uwo.ca Tue Nov 16 23:46:28 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] supravital stains References: Message-ID: <419AE5B4.D0941B55@uwo.ca> Dear Jenny Oblander, You may be way out on the wrong track, looking for a dye that does not really exist. Rhodanile blue was a product made and sold by Edward Gurr in the 1960s. It was supposedly formed by chemical combination of rhodamine B with nile blue A. For an intelligent but inconclusive discussion of this product, see the 9th edition (1977) of Conn's Biological Stains, page 411. Rhodanile blue is not mentioned in the 10th edition of Conn's because there's probably no such dye, even if some suppliers sell something with that name. Anyone intending to use a product called rhodanile blue should find and read all the references in the 9th edition of Conn's as well as Gurr's books. These books are in public and university libraries, and on the shelves of labs that contain stainthusiasts. The reference for staining Heinz bodies (here cited from Lillie's 9th edn of Conn's) is Simpson GF et al 1970. Stain Technol. 45:221-223. Look up and evaluate this paper. Did Simpson et al know what they were doing? Querky dye methods may not be the best way to stain Heinz bodies. If you can explain what they contain, you will probably get an abundance of replies from histonetters. -- ------------------------------- 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 _______________________________ Jenny Oblander wrote: > > Hi again, > Does anyone have a procedure for Rhodanile blue.I'm looking for Heinz > bodies. Thanks Jenny > > J.Oblander, HT (A.S.C.P.) > > Comparative Medicine > Oklahoma Medical Research Foundation MS#32 > 825 NE 13th St. > Okc,Ok 73104 > jenny-oblander@omrf.ouhsc.edu > 405-271-7083 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Wed Nov 17 02:36:51 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] RE: MUC6 antibody Message-ID: <263bf02630f4.2630f4263bf0@amc.uva.nl> Dear Pat, Indeed, as Gayle Callis said we did even triple staining with MUC6, MUC5AC and Helicobacter Pylori on FFPE's (see van den Brink et al, Gut (2000) 46:601-607). At that time the anti-MUC6 antibody (rabbit) was a gift, but I have seen anti-MUC6 (mouse) now available from Novocastra. Perhaps you should give it a try. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "Pat Zeitlow" Date Tue, 16 Nov 2004 10:01:56 -0600 To Subject [Histonet] MUC6 antibody Does anyone know a laboratory doing this stain on ffpe tissues? Pat Z Department of Molecular Pathology Boyce and Bynum Pathology Labs, P.C. From dmikita <@t> wmcnet.org Wed Nov 17 06:25:26 2004 From: dmikita <@t> wmcnet.org (Daryl Mikita) Date: Fri Sep 16 15:24:18 2005 Subject: [Histonet] Formalin clean-up Message-ID: We have a product that we use that solidifies, neutralize, and deodorizes 10% formalin. It is called Transform FSA from American Master*Tech. Daryl Mikita, HT(ASCP) Wyoming Medical Center 1233 E. 2nd St. Casper, WY 82601 From MAUGER <@t> email.chop.edu Wed Nov 17 07:00:56 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Tissue Transglutaminase Message-ID: Dear histonettors, Has anyone had any luck with this antibody on FFPE tissue sections? I have tried two differentAbs- one from Dako and one from chemicon with no luck yet. I have tried every retrieval method I know of. Any suggestions? Thanks to all, Jo >>> "Daryl Mikita" 11/17/04 07:25AM >>> We have a product that we use that solidifies, neutralize, and deodorizes 10% formalin. It is called Transform FSA from American Master*Tech. Daryl Mikita, HT(ASCP) Wyoming Medical Center 1233 E. 2nd St. Casper, WY 82601 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akbitting <@t> geisinger.edu Wed Nov 17 07:41:45 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Luciferase antibody Message-ID: Have you tried contacting the Vatican?............sorry that was a bad joke. :) 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 >>> 11/16/04 05:15PM >>> Hi everyone, I am looking for a nice anti-luciferase antibody for immunohistochemistry for formalin-fixed, paraffin-embedded sections of the lung. Tom Kuzniar t-kuzniar@md.northwestern.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 POWELL_SA <@t> Mercer.edu Wed Nov 17 07:59:51 2004 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] coverslipping with tape In-Reply-To: <001601c4cc35$bf41a5a0$6401a8c0@gateway> Message-ID: When that happens here, I dip both into Xylene and use a resin media to stick the cover slip back to the slide or you can use a clean slide but it works. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Brian D. Conlon Sent: Tuesday, November 16, 2004 6:41 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] coverslipping with tape hi everyone,does anyone know how to save slides that have been coverslipped with tape on a sakura coverslipper? we pulled some slides that had been filed for a couple of years and the tape is loose and the sections have adhered to the tape instead of the slide. any suggestions? thank you in advance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DOOLEEO <@t> shands.ufl.edu Wed Nov 17 08:45:51 2004 From: DOOLEEO <@t> shands.ufl.edu (Elaine Dooley) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] TFE3 Message-ID: Dear Histonet subscribers and vendors, Does anyone know of any companies that supple the antibody to "TFE3"? Also does it work on formalin fixed paraffin embedded material. Thanks in advance Elaine Dooley Shands Teaching Hospital Gainesville FL 352-265-0111 ext 72114 From eca9 <@t> georgetown.edu Wed Nov 17 09:03:21 2004 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] staining of rat gut tissue? Message-ID: <419B6839.4030107@georgetown.edu> Hi, I have a small question. Does anybody know of an antibody that will show good staining in Rat gut tissue? I just need to prove that my tissue will stain. Thanks, Eva Georgetown University From eca9 <@t> georgetown.edu Wed Nov 17 09:59:04 2004 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] staining of rat gut tissue? :explanation Message-ID: <419B7548.4070702@georgetown.edu> I don't have any particular type of staining in mind. This is the beginning of a small project and my boss just wants to make sure I can do a simple staining of the gut tissue that I fixed and had paraffin embedded before he lets me loose on the more important samples. He asked me to look into an antibody that will show good staining in the rat gut tissue on top of an H& E staining. My H& E staining looks fine. I suppose it could be seen as a control staining. Is there any particular structure (protein)in rat gut tissue that always shows up with a particular antibody? Hope this clears up some of the confusion. Thank you all for your help. Eva Georgetown University From rschoon <@t> email.unc.edu Wed Nov 17 10:19:51 2004 From: rschoon <@t> email.unc.edu (rschoon) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] staining of rat gut tissue? :explanation In-Reply-To: <419B7548.4070702@georgetown.edu> References: <419B7548.4070702@georgetown.edu> Message-ID: <419B7A27.1000506@email.unc.edu> PCNA would work. There is a high cell turnover rate in the gut. Eva C Andersson wrote: > I don't have any particular type of staining in mind. This is the > beginning of a small project and my boss just wants to make sure I can > do a simple staining of the gut tissue that I fixed and had paraffin > embedded before he lets me loose on the more important samples. He > asked me to look into an antibody that will show good staining in the > rat gut tissue on top of an H& E staining. My H& E staining looks > fine. I suppose it could be seen as a control staining. Is there any > particular structure (protein)in rat gut tissue that always shows up > with a particular antibody? Hope this clears up some of the confusion. > Thank you all for your help. > Eva > Georgetown University > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MAUGER <@t> email.chop.edu Wed Nov 17 11:12:03 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] staining of rat gut tissue? :explanation Message-ID: Hi Histonetters, Please ignore earlier question about tTG. I was confused. I meant Inhibin (beta)B. I have been trying the C-18 and G-19 from Santa Cruz with no luck on FFPE tissue. Anyone familiar with this please reply.Thanks for understanding my "senior"though I hate to admit it, moment. Jo >>> rschoon 11/17/04 11:19AM >>> PCNA would work. There is a high cell turnover rate in the gut. Eva C Andersson wrote: > I don't have any particular type of staining in mind. This is the > beginning of a small project and my boss just wants to make sure I can > do a simple staining of the gut tissue that I fixed and had paraffin > embedded before he lets me loose on the more important samples. He > asked me to look into an antibody that will show good staining in the > rat gut tissue on top of an H& E staining. My H& E staining looks > fine. I suppose it could be seen as a control staining. Is there any > particular structure (protein)in rat gut tissue that always shows up > with a particular antibody? Hope this clears up some of the confusion. > Thank you all for your help. > Eva > Georgetown University > > _______________________________________________ > Histonet mailing list > Histonet@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 jlucas <@t> hsc.wvu.edu Wed Nov 17 11:36:21 2004 From: jlucas <@t> hsc.wvu.edu (Jenny Lucas) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] To all Histotechs: Message-ID: To all Histotechs: Can someone provide me with the name of the indicator used to determine whether or not the primary staining solution used in the PAS procedure is still viable? Thanks, Jen Jenny Lucas B.S., B.A., CTBS (AATB) West Virginia University Tissue Bank Tissue Bank Associate From susan.wells <@t> bms.com Wed Nov 17 11:53:07 2004 From: susan.wells <@t> bms.com (Susan Q Wells) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] CD11b paraffin IHC Message-ID: <419B9003.5060200@bms.com> Has anyone used CD11b for formalin fixed IHC ? Thanks in advance. Sue Wells From northma <@t> ohsu.edu Wed Nov 17 11:58:47 2004 From: northma <@t> ohsu.edu (Mary North) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] To all Histotechs: Message-ID: >From Dezna Sheehan's book: Tests for Schiff reagent: Chemical(AFIP). Pour 10 ml of 37-40% formalin into a watch glass. To this add a few drops of the Schiff reagent to be tested. A good Schiff reagent will rapidly turn a red-purple color. A deteriorating Schiff reagent will give a delayed reaction and the color produced will be a deep blue-purple." Mary North OHSU, Portland, OR >>> "Jenny Lucas" 11/17/2004 9:36:21 AM >>> To all Histotechs: Can someone provide me with the name of the indicator used to determine whether or not the primary staining solution used in the PAS procedure is still viable? Thanks, Jen Jenny Lucas B.S., B.A., CTBS (AATB) West Virginia University Tissue Bank Tissue Bank Associate _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Nov 17 12:11:08 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Schiffs reagent viability test, qualitative In-Reply-To: References: Message-ID: <6.0.0.22.1.20041117105831.01b2e6a0@gemini.msu.montana.edu> 10 mls formaldehyde, I often use neutral buffered formalin and add a few drops of Schiffs reagent to this with a pasteur pipette. Use a clear glass beaker so you can see color development easily. The Schiffs reacts with the -aldehyde, and fresh, new Schiffs will turn a very bright dark, red-pink color VERY rapidly. I suggest you try this with fresh newly opened reagent to see what color you get - this will help you calibrate your eye to the color that develops. One caveat: do NOT pour used Schiffs back into the Stock solution bottle, we store used Schiffs that is contaminated with water carryover from slides (dilution factor) in a separately labeled bottle and if we reuse this, do the test first or toss used Schiffs if only using a few mls i.e. 20 mls or so. Also, if Schiffs develops a precipitate, we toss it and we never allow it to freeze. 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 Wed Nov 17 12:12:19 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] CD11b paraffin IHC In-Reply-To: <419B9003.5060200@bms.com> References: <419B9003.5060200@bms.com> Message-ID: <6.0.0.22.1.20041117111129.01b03a60@gemini.msu.montana.edu> What species? At 10:53 AM 11/17/2004, you wrote: >Has anyone used CD11b for formalin fixed IHC ? >Thanks in advance. >Sue Wells > > >_______________________________________________ >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 pruegg <@t> ihctech.net Wed Nov 17 12:13:42 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] question post for Laura Message-ID: <000c01c4ccd1$2b90fd50$83020a0a@IHCTech> Please respond directly to Laura on this bliven.laura@marshfieldclinic.org From: bliven.laura@marshfieldclinic.org Subject: Histonet Question Date: Tue, 16 Nov 2004 16:18:53 -0600 Hi Patsy, Since our IS dept. keeps deleting me from the Histonet, (I need to talk with them), could you please forward a question to the group? The question for today is: Because of new regulations, our current incinerator will be torn down in the near future. I understand that there are other ways to dispose of tissue and we'd like to know the possibilities and systems. I understand that there are expensive crematory-type and pressure cooker types of systems. Any ideas and company info will be greatly appreciated. Thanks, Laura Bliven Marshfield Laboratories Marshfield, WI 54449 bliven.laura@marshfieldclinic.org (715)387-7810 From liz <@t> premierlab.com Wed Nov 17 12:15:44 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] CD11b paraffin IHC In-Reply-To: <419B9003.5060200@bms.com> Message-ID: <000001c4ccd1$74302540$76d48a80@AMY> Sue CD11b will not work in paraffin sections. It will work in paraformaldehyde fixed frozen sections. 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 Susan Q Wells Sent: Wednesday, November 17, 2004 10:53 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] CD11b paraffin IHC Has anyone used CD11b for formalin fixed IHC ? Thanks in advance. Sue Wells _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From billingconsultants <@t> yahoo.com Wed Nov 17 13:05:44 2004 From: billingconsultants <@t> yahoo.com (Caldwell) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Cytology Technicians Message-ID: <20041117190544.75556.qmail@web54210.mail.yahoo.com> Hi everyone, Would any of you be aware of a list serv or news group for cytology technicians? Thank you in advance for your assistance. Louri Roberts Caldwell ------------------------------- Billing Consultants, LLC www.billingconsultants.net --------------------------------- Do you Yahoo!? The all-new My Yahoo! – Get yours free! From ajennings <@t> unmc.edu Wed Nov 17 13:25:24 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] subject line is important In-Reply-To: <6.0.0.22.1.20041117105831.01b2e6a0@gemini.msu.montana.edu> Message-ID: I know this comes up every once in a while...and I have seen different people, through the years, make this plea......guess it is my turn...... please make sure your 'subject' or 'reference' line in your emails reflects your question or topic. I know I miss many good questions or replies because it will say "question" or "can you help?" and god forbid the inevitable question that is sent via the "digest" mailing, it is hard to track the replies, much less find the question. I don't believe any of us have the time to cycle through all the Histonet messages all of the time so if it has something familiar to our field in the subject line it is more apt to catch our attention and get a quick reply. thank you anita From Jacqueline.Miller <@t> UTSouthwestern.edu Wed Nov 17 13:33:41 2004 From: Jacqueline.Miller <@t> UTSouthwestern.edu (Jacqueline Miller) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] weird prostate Message-ID: Hi everyone, Does anyone know what would cause the glandula of the prostate to open up wide so that they're like huge holes. My sections look like I've cut swiss cheese. Under the microscope, the basal layer is usually still present, but the epithelium is all disintegrated in the middle of what's left of the gland. Not all of my prostate samples do this--some look fine. It mostly seems to happen with larger BPH samples. My samples usually sit in formalin for at least 24 hours, but not more than 72 hours (over a weekend). Then, they sit in 70% EtOH until they are processed (for up to a week). The processing is as follows: 70% EtOH - 2 hrs 70% EtOH - 2 hrs 80% EtOH - 2 hrs 95% EtOH - 2 hrs 95% EtOH - 2 hrs 100% EtOH - 2 hrs 100% EtOH - 2 hrs 50:50 - 2 hrs xylene - 2 hrs xylene - 2 hrs paraffin - 2 hrs paraffin - 2 hrs I don't think it's a sectioning problem because if you look at the block at the right angle, you can see that the sample itself looks like swiss cheese. I appreciate any thoughts. Thanks in advance, Jacqueline Miller UT Southwestern Medical Center Dallas, TX From micro <@t> formatex.org Wed Nov 17 14:05:56 2004 From: micro <@t> formatex.org (Advanced Materials) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Invitation to submit abstract(s) to Applied Microbiology 2005 In-Reply-To: Message-ID: <000d01c4cce0$da94c520$0d001aac@PUESTO4> 1st International Conference on Environmental, Industrial and Applied Microbiology (BioMicroWorld-2005) March 15-18th 2005, Badajoz, Spain http://www.formatex.org/biomicroworld2005 Dear Colleague, further to our recent invitation to you to take part of the forthcoming BioMicroWorld-2005 International Conference, we would like to remind that if you intend to take part of it, deadline for submission of abstracts is November 22nd (next Monday). While we will be able to accept some late abstract during the whole next week, this will be possible only for POSTERS presentations. Next monday is the absolute deadline for submission of proposals for oral presentations. Below please find some important informations about the conference. All the information is being continously updated at the website http://www.formatex.org/biomicroworld2005 MAIN TOPICS Main topics of the Conference are - Environmental Microbiology, Marine Microbiology, Water/Aquatic Microbiology, Geomicrobiology - Industrial Microbiology - Future Bioindustries - Food Microbiology - Cell Engineering - Pharmaceutical Microbiology - Agriculture, Soil, Forest Microbiology - Structure and Morphogenesis - Analytical Techniques, Imaging Techniques, Microscopy - Microbial Physiology, Metabolism and Gene Expression - Microbial Biotechnology - Aerospace Microbiology, Astro(micro)biology - Quantitative Models and Bioinformatics in Microbiology - Methods in Basic and Applied Microbiology - Medical Microbiology - Microbiology Education IMPORTANT DEADLINES Submission of abstracts: November 22th, 2004 (abstracts for posters presentations will be accepted until November 27th) Submission of Full Papers for publication: On site CONFERENCE PUBLICATIONS - PROCEEDINGS 1. Book A multi-volume book entitled "Recent Advances in Multidisciplinary Applied Microbiology. Biological, Physical, Chemical and Engineering Aspects" will be published including Full papers of works (oral, posters) presented at the conference. The book will be published by an international publisher (now in negotiations with Elsevier Science), in order to give it a broad international distribution and is intended to serve as a good reference book of the state of the art of Modern Applied Microbiology as of 2005. Submission of Full Papers is optional. 2. Abstracts Book A separate Abstracts Booklet will be published with abstracts of works presented at the Conference. It will be distributed at the beginning of the conference. 3. International Journals special issues Agreements have been arranged with several international journals, in order to produce special issues based on very good papers presented at the Conference. Manuscripts must be delivered during the Conference, according to the instructions we will give in due course for each journal. All papers will be strictly reviewed and treated as regular papers. Our goal is to produce high quality and high impact special issues based on some of the best conference papers. Papers submitted for a journal special issue and rejected (for scientific/editorial/space reasons), can be considered for inclusion in the Book. Please refer to the conference website for details. Journals considering publication of special issues include Antonie van Leeuvenhoek (Springer,confirmed), Enzyme and Microbial Technology (Elsevier,confirmed), International Biodeterioration and Biodegradation (Elsevier), Colloids and Surfaces B: Biointerfaces (Elsevier,confirmed), Geomicrobiology Journal (Elsevier), FEMS Microbiology Ecology (Elsevier), Microscopy and Microanalysis (American Microscopy Society, confirmed), Micron (Elsevier). SPECIAL SESSIONS - WORKSHOPS - Workshop on Modern Microscopy Techniques in Applied Microbiology - MICROFACTORIES - Microbial Production of Chemicals and Pharmaceuticals - Workshop on Biotechnologically Relevant Enzymes and Proteins - Workshop on Studies on Extracellular Matrix: Biology and Physico/chemistry - Workshop on Biosurfactants: Purification, Mass production, Applications - Workshop on Yeast and Bacterial Flocculation: Fundamentals and Industrial interest - Worskhop on Microbial Biosensors - Methods in Cell, Proteins, Enzymes and other Biomolecules Immobilisation - Workshop on Biohydrogen - Hydrogen production by Microorganisms, as a Novel Source of Renewable Energy - Workshop on Bioremediation - Workshop on Microbial Biopolymers: Synthesis, Characterization and Applications - Methods in Cell Adhesion: Classical and Novel Methods, from Macroscopic to Nanometer scale, from Biochemistry to Nano(bio)technology PLENARY LECTURES Plenary talks include: "Biomarkers to Define Interactions in the Environment and Health" David C. White, Director of the Center for Biomarker Analysis, University of Tennessee, USA "Novel Time-resolved Fluoresecence Based Immunoassays and Real-time PCR Assays in Microbiological Applications" Timo Lovgren, Department of Biochemistry and Food Chemistry/Biotechnology, University of Turku, Finland "The Genetics and Biochemistry of Malonate and 2,4-D Biodegradation by Burkholderia cepacia Strain 2a" Ian James Bruce, Nanobiotechnology Research Group, Istituto di Scienze Chimiche, Universita degli Studi di Urbino, ITALY / School of Science, University of Greenwich, UK Alexander Steinbuchel, Institut fur Molekulare Mikrobiologie und Biotechnologie, Munster, GERMANY [title to be announced] We hope you find it interesting to present your work(s) at the Conference, and we certainly hope to receive your abstract(s) in due course. Of course every suggestion or collaboration proposals will be appreciated to make the conference reach its goals. Best wishes, Fatima Penya BioMicroWorld-2005 Secretariat Formatex Research Centre C/Zurbaran, 1 2a Planta, Office 1 06001 Badajoz, SPAIN Phone/Fax: +34 924258615 E-mail: applmicro@formatex.org From convmcm <@t> cc.usu.edu Wed Nov 17 14:07:06 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Schiffs reagent viability test, qualitative In-Reply-To: <6.0.0.22.1.20041117105831.01b2e6a0@gemini.msu.montana.edu> Message-ID: <000001c4cce1$037c4440$4a737b81@Cygnus> I use warm tap water because that's where the slides go in the staining procedure. 10% BNF is the recommended, but I've never noticed any difference with the warm tap water. Food for thought... Connie McManus Utah Veterinary Diagnostics Laboratory Utah State University Logan, UT Phone: 435/797-1891 fax: 435/797-2805 email: convmcm@cc.usu.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Wednesday, November 17, 2004 11:11 AM To: Jenny Lucas; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Schiffs reagent viability test, qualitative 10 mls formaldehyde, I often use neutral buffered formalin and add a few drops of Schiffs reagent to this with a pasteur pipette. Use a clear glass beaker so you can see color development easily. The Schiffs reacts with the -aldehyde, and fresh, new Schiffs will turn a very bright dark, red-pink color VERY rapidly. I suggest you try this with fresh newly opened reagent to see what color you get - this will help you calibrate your eye to the color that develops. One caveat: do NOT pour used Schiffs back into the Stock solution bottle, we store used Schiffs that is contaminated with water carryover from slides (dilution factor) in a separately labeled bottle and if we reuse this, do the test first or toss used Schiffs if only using a few mls i.e. 20 mls or so. Also, if Schiffs develops a precipitate, we toss it and we never allow it to freeze. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Wed Nov 17 14:12:12 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] FW: [CSH-Officers] Channel 2 news video Message-ID: <001c01c4cce1$b9799f40$83020a0a@IHCTech> Check this out one of our histotechs made TV news. Patsy -----Original Message----- From: csh-officers-bounces@neo.agsci.colostate.edu [mailto:csh-officers-bounces@neo.agsci.colostate.edu] On Behalf Of John McGinley Sent: Wednesday, November 17, 2004 11:33 AM To: csh officers Subject: [CSH-Officers] Channel 2 news video Hi, Channel 2 News shot some footage of our lab last Friday and I posted the video our website this morning (see link below). The video talks about a collaborative project looking at antioxidants in different bean varieties to see which compounds may prove helpful in preventing cancer. New strains of beans could then be genetically engineered that have a combination of helpful antioxidants. The only reason I posted this message is because there are several shots of sections being cut and how often do you get to see histotechs you know on TV. http://www.cpl.colostate.edu/press.htm (click on link labeled "WB2 News Interview") Regards, John ----------------------------------- John N. McGinley 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 _______________________________________________ csh-officers mailing list csh-officers@neo.agsci.colostate.edu http://neo.agsci.colostate.edu/mailman/listinfo/csh-officers From cottamb <@t> ohsu.edu Wed Nov 17 14:15:47 2004 From: cottamb <@t> ohsu.edu (Benjamin Cottam) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] ELF 97 kit Message-ID: does anyone have experience using the ELF 97 kit from molecular probes on central nervous system tissue? Ben Cottam OHSU From cforster <@t> umn.edu Wed Nov 17 16:09:18 2004 From: cforster <@t> umn.edu (Colleen Forster) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] double stain question Message-ID: <419BCC0E.9010106@umn.edu> Histonetters, I was asked today if it was possible to do a Trichrome and Retic on the same slide. What is anyones opinion? If someone has dared to try would you share....... Thanks, Colleen Forster U of Mn 612-626-0436 -- Outgoing mail is certified Virus Free. Checked by AVG Anti-Virus (http://www.grisoft.com). Version: 7.0.279 / Virus Database: 265.3.1 - Release Date: 11/15/2004 From e.golder-novoselsky <@t> biogenex.com Wed Nov 17 16:27:26 2004 From: e.golder-novoselsky <@t> biogenex.com (Elina Golder-Novoselsky) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] To all Histotechs: Message-ID: Elina Golder-Novoselsky, Ph.D. Manager, Inside Sales BioGenex Laboratories, Inc. 4600 Norris Canyon Road San Ramon, CA 94583 phone: (925) 866-2518 (direct) e-mail: e.golder-novoselsky@biogenex.com From jstaruk <@t> masshistology.com Wed Nov 17 17:40:54 2004 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Manual for the Fisher Histomatic Slide Stainer 172 In-Reply-To: <349C15307203B445A6B06AA6BA3D425F04AD5150@exchange.caus.dako.net> Message-ID: <000001c4ccfe$e2069c20$6501a8c0@yourw04gtxld67> Does anyone have a copy of the manual for the Fisher Histomatic Slide Stainer 172? Anyone who plans on invoicing me for this information need not reply. Thanks Jim From ernestinemiddleton <@t> yahoo.ca Wed Nov 17 18:16:11 2004 From: ernestinemiddleton <@t> yahoo.ca (Ernestine Middleton) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Aristan Stainer Message-ID: <20041118001611.17626.qmail@web51506.mail.yahoo.com> Hi; This question is primary for the aristan stainer user. Are you using warthin starry kit to stain you stomach bxs? Do you have any problem with staining the bacteria if you are using eosin for highlighting your bxs? I found that eosin or Mercurochrome prevent the bacteria from stain. Any suggestion will be appreciated. Maybe someone maybe using something different to high-light their bxs. Ernestine Middleton, Manager Montefiore Medical Center Bronx, NY 718-920-4157 --------------------------------- Post your free ad now! Yahoo! Canada Personals From JWEEMS <@t> sjha.org Wed Nov 17 19:56:30 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Aristan Stainer Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0B8E87@sjhaexc02.sjha.org> We are using it - and modifying it to stain like the Genta - alcian blue then H&E = its beautiful... j ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ernestine Middleton Sent: Wed 11/17/2004 7:16 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] Aristan Stainer Hi; This question is primary for the aristan stainer user. Are you using warthin starry kit to stain you stomach bxs? Do you have any problem with staining the bacteria if you are using eosin for highlighting your bxs? I found that eosin or Mercurochrome prevent the bacteria from stain. Any suggestion will be appreciated. Maybe someone maybe using something different to high-light their bxs. Ernestine Middleton, Manager Montefiore Medical Center Bronx, NY 718-920-4157 --------------------------------- Post your free ad now! Yahoo! Canada Personals _______________________________________________ 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 n.cragg <@t> epistem.co.uk Thu Nov 18 07:03:26 2004 From: n.cragg <@t> epistem.co.uk (Nicola) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] IHC on rat gut tissue Message-ID: <000101c4cd6e$feb3a820$72a7a8c0@epistem.local> I would try MIB5, anti-rat Ki67on your rat gut tissue - should get some nice staining. Nicola From ian.montgomery <@t> bio.gla.ac.uk Thu Nov 18 09:52:42 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Envision. Message-ID: <6.1.2.0.2.20041118154949.02d33770@udcf.gla.ac.uk> I wonder if anyone can offer any hints and tips for Dako Envision. Localisation is fine, HRP is clean but alkaline phosphatase, despite my efforts, is still giving me quite a lot of background. This manifests itself as a fine red ppt. over the specimen. Ian. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk From MAUGER <@t> email.chop.edu Thu Nov 18 10:10:28 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Envision. Message-ID: Dr. Montgomery, Have you tried adding levamisole to your chromogen? It blocks endogenous alk-phos in tissue.Dako sells it seperately from kit. I use 1 drop per ml. of chromogen, and it works quite well. Jo >>> "Ian Montgomery" 11/18/04 10:52AM >>> I wonder if anyone can offer any hints and tips for Dako Envision. Localisation is fine, HRP is clean but alkaline phosphatase, despite my efforts, is still giving me quite a lot of background. This manifests itself as a fine red ppt. over the specimen. Ian. Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Nov 18 11:20:21 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Re: fine red ppt on sections after AP Envision. In-Reply-To: References: Message-ID: <6.0.0.22.1.20041118101330.01afc110@gemini.msu.montana.edu> In addition to Joanne's reply, using PBS rinsing at chromogen steps can provide phosphate ions that will react with the AP. Either use TRIS buffered saline throughout whole staining protocol or at least do a TBS rinse x 3 before the AP chromogen step to get rid of phosphate ions. Another nice block for endogenous Alk phos is KPL's ready to use universal blocker, done at beginning of staining protocol ( just like peroxidase blocking, up front). Nice thing is you don't have to add anything to the chromogen at the end. This block nicely replaces levamisole. At 09:10 AM 11/18/2004, you wrote: >Dr. Montgomery, >Have you tried adding levamisole to your chromogen? It blocks endogenous >alk-phos in tissue.Dako sells it seperately from kit. I use 1 drop per ml. >of chromogen, and it works quite well. >Jo > > >>> "Ian Montgomery" 11/18/04 10:52AM >>> > I wonder if anyone can offer any hints and tips for Dako Envision. >Localisation is fine, HRP is clean but alkaline phosphatase, despite my >efforts, is still giving me quite a lot of background. This manifests >itself as a fine red ppt. over the specimen. >Ian. > >Dr. Ian Montgomery, >Histotechnology, >Graham Kerr Building, >Institute of Biomedical & Life Sciences, >University of Glasgow, >Glasgow, >G12 8QQ. >Tel: 0141 339 8855 >Office: 4652 >Lab: 6644. >Pager: 07625 702883 >e-mail: ian.montgomery@bio.gla.ac.uk > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis 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 ian.montgomery <@t> bio.gla.ac.uk Thu Nov 18 11:22:06 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:19 2005 Subject: Fwd: Re: [Histonet] Envision. Message-ID: <6.1.2.0.2.20041118171919.02d42310@udcf.gla.ac.uk> > >Question: what buffer are you using? If you use PBS, then you will have >phosphate ions available for AP, TRIS buffered saline is preferred with AP >protocols. Gayle, I'm using TBS as suggested by Dako. With and without Tween, but still the same result. >At 08:52 AM 11/18/2004, you wrote: >> I wonder if anyone can offer any hints and tips for Dako >> Envision. Localisation is fine, HRP is clean but alkaline phosphatase, >> despite my efforts, is still giving me quite a lot of background. This >> manifests itself as a fine red ppt. over the specimen. >>Ian. >> >>Dr. Ian Montgomery, >>Histotechnology, >>Graham Kerr Building, >>Institute of Biomedical & Life Sciences, >>University of Glasgow, >>Glasgow, >>G12 8QQ. >>Tel: 0141 339 8855 >>Office: 4652 >>Lab: 6644. >>Pager: 07625 702883 >>e-mail: ian.montgomery@bio.gla.ac.uk >> >>_______________________________________________ >>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) > Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk From ian.montgomery <@t> bio.gla.ac.uk Thu Nov 18 12:13:20 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:19 2005 Subject: Fwd: [Histonet] Re: fine red ppt on sections after AP Envision. Message-ID: <6.1.2.0.2.20041118181137.02d3f1f0@udcf.gla.ac.uk> > >In addition to Joanne's reply, using PBS rinsing at chromogen steps can >provide phosphate ions that will react with the AP. Either use TRIS >buffered saline throughout whole staining protocol or at least do a TBS >rinse x 3 before the AP chromogen step to get rid of phosphate ions. > >Another nice block for endogenous Alk phos is KPL's ready to use universal >blocker, done at beginning of staining protocol ( just like peroxidase >blocking, up front). Nice thing is you don't have to add anything to >the chromogen at the end. This block nicely replaces levamisole. KPL, you've got me there. Where do I buy this universal blocker. Ian >At 09:10 AM 11/18/2004, you wrote: >>Dr. Montgomery, >>Have you tried adding levamisole to your chromogen? It blocks endogenous >>alk-phos in tissue.Dako sells it seperately from kit. I use 1 drop per >>ml. of chromogen, and it works quite well. >>Jo Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07625 702883 e-mail: ian.montgomery@bio.gla.ac.uk From pruegg <@t> ihctech.net Thu Nov 18 13:12:01 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] antibodies Message-ID: <001701c4cda2$7b6ccb20$83020a0a@IHCTech> I am looking for antibody recommendations that work on ffpe mouse tissues for: macrophages dendritic cells From kerry.l.crabb <@t> gsk.com Thu Nov 18 13:50:40 2004 From: kerry.l.crabb <@t> gsk.com (kerry.l.crabb@gsk.com) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] 2005 NSH S/C Speaker Proposal Message-ID: The deadline (December 1st) is drawing near for submitting a speaker proposal for the 2005 NSH S/C to be held in Fort Lauderdale, FL. If you are interested, go to the NSH web site (www.nsh.org). You'll see the convention logo for the meeting in Florida on the home page. Click in the logo area where it says to "click here to submit a your proposal". This is a new program that is more user friendly. From ashelynne2 <@t> yahoo.com Thu Nov 18 14:15:51 2004 From: ashelynne2 <@t> yahoo.com (linda cobb) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] concerning special stain controls (not immuno;s) Message-ID: <20041118201551.12703.qmail@web51707.mail.yahoo.com> Note: forwarded message attached. __________________________________ Do you Yahoo!? The all-new My Yahoo! - Get yours free! http://my.yahoo.com From liz <@t> premierlab.com Thu Nov 18 15:34:22 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] antibodies In-Reply-To: <001701c4cda2$7b6ccb20$83020a0a@IHCTech> Message-ID: <000c01c4cdb6$5ec7a260$76d48a80@AMY> Patsy F4/80 will stain mouse macrophages, both fixed (kupfer) and mobile macrophages, I would suspect they would also stain the macrophage dendritic cells. The antibody from Serotec will work on formalin fixed tissue with proteinase K digestion. There might be an antibody out there that is more specific than this one, but I'm not aware of any. 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 Patsy Ruegg Sent: Thursday, November 18, 2004 12:12 PM To: 'Histonet' Subject: [Histonet] antibodies I am looking for antibody recommendations that work on ffpe mouse tissues for: macrophages dendritic cells _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Thu Nov 18 15:51:01 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] frozen section staining Message-ID: Hello, I have a Mohs surgeon that is doing frozens at a different facility and the stain is not looking like mine. She brought a slide to me and it appears to look like the collagen is "cob webs." Medium/ bright pink. And the tumor is very dark purple...any ideas what could be causing this? Robyn OHSU Portland, Or From DWilliams <@t> ciit.org Thu Nov 18 15:55:28 2004 From: DWilliams <@t> ciit.org (Delorise Williams) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Bodian silver stain Message-ID: <12816CB59E68184F9F5F28063E68B04701BF4262@xsrvr.ciit.org> Hi Everyone, Would like to please talk to someone that has had good results with the Bodian stain. Also would like to know the most common stain or method being used in the labs today to demonstrate nerve fibers, endings and neurofibrils? I work in research primarily with animal tissue but presently am working with human nasal turbinate tissue. Delorise Williams CIIT Centers for Health Research PO Box 12137 Research Triangle Park, NC 27709 (919) 558-1200 Voice Mail-(919) 558-1252 Fax-(919) 558-1300 From sagitalcuts <@t> yahoo.com Thu Nov 18 15:58:07 2004 From: sagitalcuts <@t> yahoo.com (Yolanda Maldonado) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Average cutting of blocks Message-ID: <20041118215807.39479.qmail@web53502.mail.yahoo.com> Hello everyone: This is a question for lab managers and supervisors I would like to know what would be an average number of blocks that a Histotechnician can handle on a 7 hour working day; from embedding to coverslipping (only coverslipping is done on machine) Has anyone done a study on this? I should say that my lab works mostly with animal tissue of which, 50% is complete mice necropsies (i.e. small pieces of tissue -sometimes up to 6 organs in one cassette), the rest is usually only one piece of tissue per cassette. One H&E/cassette I would appreciate any input on this. Thanks in advance Patricia Maldonado HT (ASCP) MSKCC __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From vazquezr <@t> ohsu.edu Thu Nov 18 17:08:39 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Average cutting of blocks Message-ID: Yolanda, I am not a lab manager or supervisor, but I can tell you from experience as a technician. I could probably do 150 myself or more depending on the tissue. Robyn OHSU Portland, Or >>> Yolanda Maldonado 11/18/2004 1:58:07 PM >>> Hello everyone: This is a question for lab managers and supervisors I would like to know what would be an average number of blocks that a Histotechnician can handle on a 7 hour working day; from embedding to coverslipping (only coverslipping is done on machine) Has anyone done a study on this? I should say that my lab works mostly with animal tissue of which, 50% is complete mice necropsies (i.e. small pieces of tissue -sometimes up to 6 organs in one cassette), the rest is usually only one piece of tissue per cassette. One H&E/cassette I would appreciate any input on this. Thanks in advance Patricia Maldonado HT (ASCP) MSKCC __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emry <@t> u.washington.edu Thu Nov 18 17:21:48 2004 From: emry <@t> u.washington.edu (Trisha Emry) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] picro-indigo-carmen Message-ID: Hi, I did this in the past, but can't find a protocol on it now. If any of you still have this in your files, please, send me a copy. Thanks, Trisha U of WA From asmith <@t> mail.barry.edu Thu Nov 18 17:24:15 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Bodian silver stain Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3C42@exchsrv01.barrynet.barry.edu> Personally, I don't like the Bodian stain. For most nerves, the Holmes technique works well on the first try (Kiernan, HISTOLOGICAL AND HISTOCHEMICAL METHODS, 3rd ed., p. 366). You cannot beat Kiernan's physical developer method for really tiny nerves (Kiernan, HISTOLOGICAL AND HISTOCHEMICAL METHODS, 3rd ed., p. 371). It can detect single axons. Kiernan's method takes practice, but it is worth learning. After 3 tries, you should have mastered it. 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 Delorise Williams Sent: Thursday, November 18, 2004 4:55 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bodian silver stain Hi Everyone, Would like to please talk to someone that has had good results with the Bodian stain. Also would like to know the most common stain or method being used in the labs today to demonstrate nerve fibers, endings and neurofibrils? I work in research primarily with animal tissue but presently am working with human nasal turbinate tissue. Delorise Williams CIIT Centers for Health Research PO Box 12137 Research Triangle Park, NC 27709 (919) 558-1200 Voice Mail-(919) 558-1252 Fax-(919) 558-1300 _______________________________________________ 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 mprice26 <@t> juno.com Thu Nov 18 18:22:49 2004 From: mprice26 <@t> juno.com (marsha r price) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] RE: 2 questions Message-ID: <20041118.182249.2880.1.mprice26@juno.com> Histonetters, I have 2 different questions for you. #1. Is anyone using the CDX2 antibody and whose are you using? #2. Is there a published document on the cost of routine histology per block from start to finish? Thank you. Marsha Price ________________________________________________________________ Juno Platinum $9.95. Juno SpeedBand $14.95. Sign up for Juno Today at http://www.juno.com! Look for special offers at Best Buy stores. From jstaruk <@t> masshistology.com Thu Nov 18 18:32:55 2004 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] antibodies In-Reply-To: <000c01c4cdb6$5ec7a260$76d48a80@AMY> Message-ID: <000601c4cdcf$50c18280$6501a8c0@yourw04gtxld67> We've also used the same antibody, same pre-treatment at a 1:50 dilution with very nice results. Jim ___________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Thursday, November 18, 2004 4:34 PM To: 'Patsy Ruegg'; 'Histonet' Subject: RE: [Histonet] antibodies Patsy F4/80 will stain mouse macrophages, both fixed (kupfer) and mobile macrophages, I would suspect they would also stain the macrophage dendritic cells. The antibody from Serotec will work on formalin fixed tissue with proteinase K digestion. There might be an antibody out there that is more specific than this one, but I'm not aware of any. 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 Patsy Ruegg Sent: Thursday, November 18, 2004 12:12 PM To: 'Histonet' Subject: [Histonet] antibodies I am looking for antibody recommendations that work on ffpe mouse tissues for: macrophages dendritic cells _______________________________________________ Histonet mailing list Histonet@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 jstaruk <@t> masshistology.com Thu Nov 18 18:50:22 2004 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Average cutting of blocks In-Reply-To: <20041118215807.39479.qmail@web53502.mail.yahoo.com> Message-ID: <000001c4cdd1$c3904f10$6501a8c0@yourw04gtxld67> I did a time study on myself recently over a period of 1 week (approximately 600 specimens). This included cassettes with single tissues (surgicals) and multiple tissues (necropsies). Here are the averages: To embed and "clean" each block, 56 seconds To "face" each block, 34 seconds To write out the slide and cut one block (mostly one section), 113 seconds To manually coverslip each stained slide, 26 seconds This averaged out to 15.75 slides per hour. This did not include staining time. Jim ___________________ Jim Staruk Mass Histology Service www.masshistology.com Subject: [Histonet] Average cutting of blocks Hello everyone: This is a question for lab managers and supervisors I would like to know what would be an average number of blocks that a Histotechnician can handle on a 7 hour working day; from embedding to coverslipping (only coverslipping is done on machine) Has anyone done a study on this? I should say that my lab works mostly with animal tissue of which, 50% is complete mice necropsies (i.e. small pieces of tissue -sometimes up to 6 organs in one cassette), the rest is usually only one piece of tissue per cassette. One H&E/cassette I would appreciate any input on this. Thanks in advance Patricia Maldonado HT (ASCP) MSKCC From tahseen <@t> brain.net.pk Thu Nov 18 15:01:06 2004 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] To all Histotechs: References: Message-ID: <006901c4cdb1$ba8c9f60$972bfea9@m7c0y4> Jenny Lucas What is your actual problem? There are two type of Schiff reagent in the market one of them ( 9033 Merck, 500ml ) will use for PAS Staining, and other one ( lit pack cat # I forget ) will use for Electrophoreses. Muhammad Tahseen Histology Supervisor SKMCH & RC Lahore Pakistan. ----- Original Message ----- From: Jenny Lucas To: Sent: Thursday, November 18, 2004 2:36 AM Subject: [Histonet] To all Histotechs: > To all Histotechs: > > Can someone provide me with the name of the indicator used to determine > whether or not the primary staining solution used in the PAS procedure > is still viable? > > Thanks, > Jen > > Jenny Lucas B.S., B.A., CTBS (AATB) > West Virginia University Tissue Bank > Tissue Bank Associate > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From c.m.vanderloos <@t> amc.uva.nl Fri Nov 19 02:33:43 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] RE: Envision-AP Message-ID: <1f4b001f51d8.1f51d81f4b00@amc.uva.nl> Ian, I was thinking the same way as Gayle did. But apparently this wasn't the problem. I wondered what AP chromogen you are using? I have very good results using the newest Dako Liquid Permanent Red substrate kit. The KPL universal blocker Gayle suggested I don't know. However, I am familiar with a similar Dako product called Dual Endogenous Enzyme Block. This blocker also applied before the IHC procedure also effectively destroyed some of my epitopes resulting in a bit too clean staining result indeed.... So be careful with this stuff. Furthermore, I am using PowerVision-AP with great results. Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 ----- Original Message ----- >From Ian Montgomery Date Thu, 18 Nov 2004 17:22:06 +0000 To histonet@lists.utsouthwestern.edu Subject Fwd: Re: [Histonet] Envision. > >Question: what buffer are you using? If you use PBS, then you will have >phosphate ions available for AP, TRIS buffered saline is preferred with AP >protocols. Gayle, I'm using TBS as suggested by Dako. With and without Tween, but still the same result. >At 08:52 AM 11/18/2004, you wrote: >> I wonder if anyone can offer any hints and tips for Dako >> Envision. Localisation is fine, HRP is clean but alkaline phosphatase, >> despite my efforts, is still giving me quite a lot of background. This >> manifests itself as a fine red ppt. over the specimen. >>Ian. >> >>Dr. Ian Montgomery, >>Histotechnology, >>Graham Kerr Building, >>Institute of Biomedical & Life Sciences, >>University of Glasgow, >>Glasgow, From rschoon <@t> email.unc.edu Fri Nov 19 05:48:19 2004 From: rschoon <@t> email.unc.edu (rschoon) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] RE: Envision-AP In-Reply-To: <1f4b001f51d8.1f51d81f4b00@amc.uva.nl> References: <1f4b001f51d8.1f51d81f4b00@amc.uva.nl> Message-ID: <419DDD83.40005@email.unc.edu> Ian, I've been using the Dako Envision AP kit for years but (after much trial and error), I have use the New Fuchsin Kit from BioGenex as my label. Works great when used properly and it is permanent. Avoid xylene after dehydration, instead usa a non-aliphatic clearing agent (I use Richard Allan's Clear Rite 3). Coverslip with your choice of media. The results are a nice bright red which doesn't seam to fade at least not in the last 7-8 years that I've used it. Ive not used the "PowerVision that Chris mentioned and so can't comment on it. I would like some information on it (better yet a trial). Robeert Schoonhoven UNC-CH C.M. van der Loos wrote: >Ian, >I was thinking the same way as Gayle did. But apparently this wasn't the problem. I wondered what AP chromogen you are using? I have very good results using the newest Dako Liquid Permanent Red substrate kit. >The KPL universal blocker Gayle suggested I don't know. However, I am familiar with a similar Dako product called Dual Endogenous Enzyme Block. This blocker also applied before the IHC procedure also effectively destroyed some of my epitopes resulting in a bit too clean staining result indeed.... So be careful with this stuff. >Furthermore, I am using PowerVision-AP with great results. >Hope this helps. > >Chris van der Loos, PhD >Dept. of Pathology >Academical Medical Center >Meibergdreef 9 >NL-1105 AZ Amsterdam >The Netherlands >phone: +31 20 5665631 > >----- Original Message ----- >>From Ian Montgomery >Date Thu, 18 Nov 2004 17:22:06 +0000 >To histonet@lists.utsouthwestern.edu >Subject Fwd: Re: [Histonet] Envision. > > > >>Question: what buffer are you using? If you use PBS, then you will have >>phosphate ions available for AP, TRIS buffered saline is preferred with AP >>protocols. >> >> >Gayle, > I'm using TBS as suggested by Dako. With and without Tween, but >still the same result. > > > > > >>At 08:52 AM 11/18/2004, you wrote: >> >> >>> I wonder if anyone can offer any hints and tips for Dako >>>Envision. Localisation is fine, HRP is clean but alkaline phosphatase, >>>despite my efforts, is still giving me quite a lot of background. This >>>manifests itself as a fine red ppt. over the specimen. >>>Ian. >>> >>>Dr. Ian Montgomery, >>>Histotechnology, >>>Graham Kerr Building, >>>Institute of Biomedical & Life Sciences, >>>University of Glasgow, >>>Glasgow, >>> >>> > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From jnocito <@t> satx.rr.com Fri Nov 19 06:26:13 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Jean Mitchell Message-ID: <00de01c4ce32$f5cd0fc0$286ece44@yourxhtr8hvc4p> Jean, please contact me. I have a brainy question. Joe the Toe From JosefaNava <@t> texashealth.org Fri Nov 19 07:24:02 2004 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] good clone for TTF1 Message-ID: <2C515C1049EAF5459EFD8C9B929078A4194455@phdex03.txhealth.org> Hello Everyone, Can someone tell me what TTF-1 clone are you using and the protocol that works best for Ventana. I thank you for your help. Josie Nava Presbyterian Hospital- Dallas The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From CBWright <@t> nmcsd.med.navy.mil Fri Nov 19 09:24:13 2004 From: CBWright <@t> nmcsd.med.navy.mil (Wright, Clarissa B CIV) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] good clone for TTF1 Message-ID: <4D840F2F36912841BD8B2484E8C683930AAEB7A7@nmc-sdca-exch02.med.navy.mil> Hi Josie, The clone I am using for TTF-1 is (8G7G3/1) from Cell Marque. Protocol for benchmark: Deparaffinization CC1-mild Primary antibody 32 min hematoxylin 4min Clarissa Wright Naval Medical Center San Diego -----Original Message----- From: Nava, Josefa [mailto:JosefaNava@texashealth.org] Sent: Friday, November 19, 2004 5:24 AM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] good clone for TTF1 Hello Everyone, Can someone tell me what TTF-1 clone are you using and the protocol that works best for Ventana. I thank you for your help. Josie Nava Presbyterian Hospital- Dallas The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This document may contain information covered under the Privacy Act, 5 USC 552(a), Health Insurance Portability and Accountability Act, Public Law 104-191, and DoD Directive 6025.18. It must be protected in accordance with those provisions. From juan.gutierrez <@t> christushealth.org Fri Nov 19 09:28:50 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] good clone for TTF1 Message-ID: We use Biocare's clone 8G7G3/1 at 1:50 with iVIEW. Extended CC1 Primary for 32min A/B Block Comes out beautiful every time. Good luck. By the way, we use the old Benchmarks, not the XT. 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: Nava, Josefa [mailto:JosefaNava@texashealth.org] Sent: Friday, November 19, 2004 7:24 AM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] good clone for TTF1 Hello Everyone, Can someone tell me what TTF-1 clone are you using and the protocol that works best for Ventana. I thank you for your help. Josie Nava Presbyterian Hospital- Dallas The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ccrowder <@t> mail.vetmed.lsu.edu Fri Nov 19 10:45:05 2004 From: ccrowder <@t> mail.vetmed.lsu.edu (Cheryl Crowder) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Thanks for the info Message-ID: I want to thank all of you who gave me replies about neutrophils. I have begun the project - just have to wait for results. However, my biggest thanks comes to all of you. The researcher involved in this project was so impressed because of the group we have, who actually talk to each other and share information. He made the comment that in the field of research rarely to people talk for fear of "stealing" an idea. So thanks to the Histonet! Cheryl Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA 70803 225-578-9734 FAX: 225-578-9720 From Nancy.Lowen <@t> med.va.gov Fri Nov 19 11:08:39 2004 From: Nancy.Lowen <@t> med.va.gov (Nancy.Lowen@med.va.gov) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] unsubscribe Message-ID: From onep00 <@t> hotmail.com Fri Nov 19 12:42:58 2004 From: onep00 <@t> hotmail.com (Pam O'Neill) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] acetic zinc formalin(AZF) and immunohistochemistry Message-ID: Help! Our hospital is phasing out b-5 fixative because of it's mercury content and hazard to the environment. We have started testing substitutes, one of them the fixing agent, AZF(see subject). It works really well on hematoxylin and eosin(H&E) stained sections of lymph node and bone marrow biopsies. However, we are just starting to test on immunohistochemical reagents with much less success. Our bone marrows which are decaled in RDO decal solution and fixed with AZF light up everything with nonspecific staining (Kappa and Lambda). We have a Dako autostainer and a Ventana LT and XT. The same problem exists on both these machines. The only time we tested a lymph node fixed in AZF we had great success with CD-30(Ki-1) but negative staining on the T and B cell antibodies tested on the Ventana. I am continuing to test this product as I write this. Anyone who is currently using this fixative, available from Newcomer Supply, and has worked out the details on the Ventana or Dako stainers please let me know. Pam O'Neill , St Vincent Mercy Medical Center, Toledo, Ohio USA ; ; From juan.gutierrez <@t> christushealth.org Fri Nov 19 13:58:24 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] acetic zinc formalin(AZF) and immunohistochemistry Message-ID: Hi Pam. We are using B-Plus fixative from BBC with great results, are you able to try something different? The only adjustment we had to do was reducing the time in the hematoxylin by almost half. Our immunos look great on the Benchmarks. 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: Pam O'Neill [mailto:onep00@hotmail.com] Sent: Friday, November 19, 2004 12:43 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] acetic zinc formalin(AZF) and immunohistochemistry Help! Our hospital is phasing out b-5 fixative because of it's mercury content and hazard to the environment. We have started testing substitutes, one of them the fixing agent, AZF(see subject). It works really well on hematoxylin and eosin(H&E) stained sections of lymph node and bone marrow biopsies. However, we are just starting to test on immunohistochemical reagents with much less success. Our bone marrows which are decaled in RDO decal solution and fixed with AZF light up everything with nonspecific staining (Kappa and Lambda). We have a Dako autostainer and a Ventana LT and XT. The same problem exists on both these machines. The only time we tested a lymph node fixed in AZF we had great success with CD-30(Ki-1) but negative staining on the T and B cell antibodies tested on the Ventana. I am continuing to test this product as I write this. Anyone who is currently using this fixative, available from Newcomer Supply, and has worked out the details on the Ventana or Dako stainers please let me know. Pam O'Neill , St Vincent Mercy Medical Center, Toledo, Ohio USA ; ; _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Fri Nov 19 14:01:10 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Parvovirus Message-ID: Is there a reference lab out there doing this marker? I will be developing it in my lab, but I need to run a case fairly quick before I can get my antibody in. Thanks netters. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! From mprice26 <@t> juno.com Sat Nov 20 13:14:12 2004 From: mprice26 <@t> juno.com (marsha r price) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Re: Thanks and what about Question #2? Message-ID: <20041120.131412.3316.4.mprice26@juno.com> Dear histonetters, Thanks to all who replied to my question about CDX2 antibody. Does anyone know of a survey or article that gives the breakdown of how much it costs to make an H&E slide from start to finish? Thank you. Marsha On Fri, 19 Nov 2004 12:52:02 -0700 "Johnson, Sandra J. (MCS)" writes: > We use the Biogenex CDX2 concentrate diluted to 1:100 (with citrate > HIER). It looks fabulous in colon carcinoma(nuclear stain) and > doesn't stain mesothelioma, breast, prostate, or lung cancers. > > Sandy Johnson, HTL > IHC Lead Tech, Histology > Mayo Clinic Scottsdale > > -----Original Message----- > From: marsha r price [mailto:mprice26@juno.com] > Sent: Thursday, November 18, 2004 17:23 PM > To: histonet@pathology.swmed.edu > Subject: [Histonet] RE: 2 questions > > > Histonetters, > I have 2 different questions for you. > > #1. Is anyone using the CDX2 antibody and whose are you using? > > #2. Is there a published document on the cost of routine histology > per > block from start to finish? > > Thank you. > > Marsha Price > > ________________________________________________________________ > Juno Platinum $9.95. Juno SpeedBand $14.95. > Sign up for Juno Today at http://www.juno.com! > Look for special offers at Best Buy stores. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ________________________________________________________________ Juno Platinum $9.95. Juno SpeedBand $14.95. Sign up for Juno Today at http://www.juno.com! Look for special offers at Best Buy stores. From lpwenk <@t> sbcglobal.net Sat Nov 20 19:42:26 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] HT/HTL EXAM RESULTS Message-ID: <000d01c4cf6b$5c0e2e20$13f7ff44@domainnotset.invalid> If you are wondering what the statistics/pass rate for the Jan-June 2004 HT and HTL exams - read further. If bored already, delete quickly. Information taken from the ASCP Board of Registry newsletter "BOR Newsletter" Fall 2004. Minimum pass score is 400, for both the written (MCQ = multiple choice question) and the practical. Reminder - both the written and the practical must be passed, to be certified. HT-MCQ # taking = 460 Mean score = 396 Range of scores = 100-788 Total pass = 230 (50%) HT-PRACTICAL # taking = 375 Mean score = 469 Range of scores = 189-889 Total pass = 283 (75%) HT-COMBINED (MCQ and Practical) # taking = 480 Total pass = 234 (49%) - - - - - HTL-MCQ # taking = 80 Mean score = 429 Range of scores = 231-725 Total pass = 47 (59%) HTL-PRACTICAL # taking = 65 Mean score = 498 Range of scores = 328-712 Total pass = 58 (89%) HTL-COMBINED (MCQ and Practical) # taking = 78 Total pass = 50 (64%) - - - - - - HT - first year certification offered = 1948 - total certified to date = 19,085 HTL - first year certification offered = 1980 - total certified to date = 2,178 - - - - Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 From contact <@t> excaliburpathology.com Sun Nov 21 11:10:28 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] 2 Ventanas for sale Message-ID: <20041121171028.30278.qmail@web50302.mail.yahoo.com> Hello, I have 2 Ventana automatic immunostainers for sale. One is a Nexus and the other is an ES. Both are in very good working order and are offered at a very reasonable price. Please feel free to contact me with any questions you may have. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From skippy_ette <@t> sbcglobal.net Sun Nov 21 18:12:41 2004 From: skippy_ette <@t> sbcglobal.net (Lori Cummings) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Fwd: Looking for Kim Peterson, MN Message-ID: <20041122001241.35021.qmail@web80409.mail.yahoo.com> Note: forwarded message attached. ===== Lori A. Cummings BS, HT(ASCP), QIHC Pathology Equipment Xchange, Inc. 1643 N. Alpine Rd., Suite 104, PMB 123 Rockford, IL 61107 (815)395-8311 office (630)417-8208 cellular lori@pathx.com From dcrippen <@t> buckinstitute.org Mon Nov 22 12:23:54 2004 From: dcrippen <@t> buckinstitute.org (Danielle Crippen) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] freezing mouse eyes Message-ID: <4AA34A707932424EBE2D973764D226A901D92FBD@inverness.buckcenter.org> Dear all, I have several sets of mouse eyes from perfused mice which have been in 4% PFA over the weekend. The investigator has just informed me this morning that she needs these eyes frozen. Can anyone with experience please elucidate what you would do in this scenario?? Should I cryoprotect? If so, should I use a sucrose gradient: 10%, 20%, 30%?? Also, how long should each of these steps be?? At our institution, the most common freezing method is by floating the tissue in a foil boat in liquid nitrogen. Any better suggestions?? Many many thanks to all the experts who I'm hoping will respond:-) Cheers, Danielle Crippen From pruegg <@t> ihctech.net Mon Nov 22 12:40:54 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] how to identify what cell type is caspase3+ Message-ID: <000901c4d0c2$cfaada10$83020a0a@IHCTech> I have a question from an investigator who is looking at rat lungs for apoptosis by using cleaved caspase 3 IHC and then wants to know what cell type is under going apoptosis (are they endothelial cells or a mix of different cells? how can they demonstrate the cell type that is positive for CC3 since the morphology changes when this happens) they do not appear to be smooth muscle cells since there is no positivity around the arteries. Should be be doing double IHC labeling say CC3 and CD31 or F8 together in the same section? Will the cell undergoing apoptosis still be expressing these other markers? Patsy From Luis.Chiriboga <@t> med.nyu.edu Mon Nov 22 13:08:20 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] RE: [IHCRG] how to identify what cell type is caspase3+ In-Reply-To: <000901c4d0c2$cfaada10$83020a0a@IHCTech> Message-ID: Hi Patsy Yes, That's exactly the approach we use.....double labeling should catch all CC3/Surface Marker+ cells except for those that have lost significant fractions of their cell membrane. Luis ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 -----Original Message----- From: ihcrg-bounces@neo.agsci.colostate.edu [mailto:ihcrg-bounces@neo.agsci.colostate.edu]On Behalf Of Patsy Ruegg Sent: Monday, November 22, 2004 1:41 PM To: 'Histonet' Cc: IHCRG@neo.agsci.colostate.edu; jenrueyt@hotmail.com Subject: [IHCRG] how to identify what cell type is caspase3+ I have a question from an investigator who is looking at rat lungs for apoptosis by using cleaved caspase 3 IHC and then wants to know what cell type is under going apoptosis (are they endothelial cells or a mix of different cells? how can they demonstrate the cell type that is positive for CC3 since the morphology changes when this happens) they do not appear to be smooth muscle cells since there is no positivity around the arteries. Should be be doing double IHC labeling say CC3 and CD31 or F8 together in the same section? Will the cell undergoing apoptosis still be expressing these other markers? Patsy _______________________________________________ IHCRG mailing list IHCRG@neo.agsci.colostate.edu http://neo.agsci.colostate.edu/mailman/listinfo/ihcrg From eca9 <@t> georgetown.edu Mon Nov 22 13:18:48 2004 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] sizes of coverslipping glass Message-ID: <41A23B98.20002@georgetown.edu> Hi, Have a small question about glass coverslips. Right now I am using a coverslip that is 24mm x 50mm. We would however need a coverslip that is large enough to cover the entire clear glass part of the slide. Is there anywhere I can get a hold of a coverslip that is 24mm x about 55mm? Thank you all in advance, Eva Andersson Georgetown University From gcallis <@t> montana.edu Mon Nov 22 13:37:56 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] sizes of coverslipping glass In-Reply-To: <41A23B98.20002@georgetown.edu> References: <41A23B98.20002@georgetown.edu> Message-ID: <6.0.0.22.1.20041122123656.01b27410@gemini.msu.montana.edu> Surgipath has these and they are superb!! I strongly suggest getting 1 1/2 thickness, correct for doing photomic work. At 12:18 PM 11/22/2004, you wrote: >Hi, >Have a small question about glass coverslips. Right now I am using a >coverslip that is 24mm x 50mm. >We would however need a coverslip that is large enough to cover the entire >clear glass part of the slide. Is there anywhere I can get a hold of a >coverslip that is 24mm x about 55mm? >Thank you all in advance, >Eva Andersson >Georgetown University > >_______________________________________________ >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 akbitting <@t> geisinger.edu Mon Nov 22 13:53:11 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Whole mount prostate specimens Message-ID: I have been asked to research what type microtome, slides, coverslips, knives I will need to begin cutting whole mount prostates. If anyone out there is doing this in the clinical hospital setting, could you help me out. Any procedures, hints, resources,....anything will help. What I'd really like to do is see someone actually cutting these type specimens. Get some one-on-one training. I'm in central Pennsylvania if anyone in the area wants to offer me some personal instruction. Thanks folks! Angie 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 contact <@t> excaliburpathology.com Mon Nov 22 14:24:57 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Freezing mouse eyes Message-ID: <20041122202457.90161.qmail@web50310.mail.yahoo.com> Have you ever sectioned eyes before? What part of the eye is being investigated? Why are they needing to do frozens, if they have been fixed? Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From kzhong888 <@t> yahoo.com Mon Nov 22 14:40:17 2004 From: kzhong888 <@t> yahoo.com (dfs dsaf) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Looking for Mark Tarango Message-ID: <20041122204017.95501.qmail@web54409.mail.yahoo.com> Hi Mark Tarango. please call me at 626 432 5032. Kirk Zhong __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From gcallis <@t> montana.edu Mon Nov 22 14:54:55 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Coverslips 24 X 55 size also available from Fisher and VWR Message-ID: <6.0.0.22.1.20041122135232.01b2acf0@gemini.msu.montana.edu> My Erie rep gave me the catalog numbers for Fisher 24 x 55 1 1/2 thickness #22291815 OR 24 X 44 1 thickness #1254418 VWR also will be able to supply Erie Scientific coverglass Nice to have choices 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 cfavara <@t> niaid.nih.gov Mon Nov 22 15:13:37 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] freezing mouse eyes Message-ID: I would ask what the investigator hopes to learn from the frozen specimens, and how they would like that accomplished. I have done staining both chemical and immuno on fresh frozen, fixed cryoprotected frozen, and paraffin embedded mouse eyes. I find the most difficult area to be the orientation for embedding. 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: Danielle Crippen [mailto:dcrippen@buckinstitute.org] Sent: Monday, November 22, 2004 11:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] freezing mouse eyes Dear all, I have several sets of mouse eyes from perfused mice which have been in 4% PFA over the weekend. The investigator has just informed me this morning that she needs these eyes frozen. Can anyone with experience please elucidate what you would do in this scenario?? Should I cryoprotect? If so, should I use a sucrose gradient: 10%, 20%, 30%?? Also, how long should each of these steps be?? At our institution, the most common freezing method is by floating the tissue in a foil boat in liquid nitrogen. Any better suggestions?? Many many thanks to all the experts who I'm hoping will respond:-) Cheers, Danielle Crippen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Nov 22 16:02:30 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:19 2005 Subject: Marking ink for maintaining orientation RE: [Histonet] freezing mouse eyes In-Reply-To: References: Message-ID: <6.0.0.22.1.20041122144525.01b20670@gemini.msu.montana.edu> I agree that orientation is always a problem with small samples. We have used green (color is easy to see) marking ink to maintain orientation of samples for snap freezing while using the petri dish floating in liquid nitrogen snap freezing method. Care must be taken NOT to be heavy handed with the dye as too much will defeat orientation purpose when a tiny sample is completely smeared with green! An extremely fine brush helps with dye application along with some type of magnifier - we use eye glass loupes attached to our spectacles. Ink can be used on any of the type of samples you mention. Using very fine forceps, one can orient the eye to bottom of plastic Tissue tek cryomold as the OCT begins to freeze (turns white) - with practice, it can be done. We do this with cross sections of very tiny murine spinal cord and maintain orientation without problems. Eyes are larger in diameter than some of the cord cross sections. With practice, one should be able to maintain orientation with any type sample using careful inking. At 02:13 PM 11/22/2004, you wrote: >I would ask what the investigator hopes to learn from the frozen specimens, >and how they would like that accomplished. I have done staining both >chemical and immuno on fresh frozen, fixed cryoprotected frozen, and >paraffin embedded mouse eyes. I find the most difficult area to be the >orientation for embedding. > >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: Danielle Crippen [mailto:dcrippen@buckinstitute.org] >Sent: Monday, November 22, 2004 11:24 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] freezing mouse eyes > >Dear all, > >I have several sets of mouse eyes from perfused mice which have been in 4% >PFA over the weekend. The investigator has just informed me this morning >that she needs these eyes frozen. Can anyone with experience please >elucidate what you would do in this scenario?? Should I cryoprotect? If >so, should I use a sucrose gradient: 10%, 20%, 30%?? Also, how long should >each of these steps be?? At our institution, the most common freezing >method is by floating the tissue in a foil boat in liquid nitrogen. Any >better suggestions?? > >Many many thanks to all the experts who I'm hoping will respond:-) > >Cheers, > >Danielle Crippen > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis 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 yej2 <@t> leicester.ac.uk Tue Nov 23 05:55:10 2004 From: yej2 <@t> leicester.ac.uk (El Jai, Y.) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Offtopic - Microscope for kids Message-ID: <1F2CE8D4B0195E488213E8B8CCF71486034C96B9@Saffron.cfs.le.ac.uk> ---------------------------- Yasmine El Jai Lab 205 Dept of Biochemistry University Road University of Leicester UK +44.1162523475 ---------------------------- From c.johnson <@t> qub.ac.uk Tue Nov 23 06:17:43 2004 From: c.johnson <@t> qub.ac.uk (Chris Johnson) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] catecholamine/glyoxylic acid fluoresence Message-ID: <000001c4d156$6f7fb6d0$b53f758f@cjohnson> Dear All, I have recently been trying to show up sympathetic innervation of blood vessels (noradrenaline/norepinephrine. I have used rat tail artery (i.e., good sympathetic innervation) incubated in phosphate buffer and 2 % glyoxylic acid pH?d to 7.4 for 90 minutes followed 4 minutes at 100 ?C. At best I have seen very faint fluorescence with the very slightest hint of a nerve plexus. I don?t think it is the processing as I have used it successfully before and it seems to be fairly robust. I suspect it is our filtering, but on looking at the literature, I?m not sure. I am using Excitation 380 ? 400 nm Dichromatic 440 nm high pass Emission 455 ? 485 nm What are the emission characteristics of the fluorophore from this reaction? What am I doing wrong?!!!! Any suggestions would be greatly appreciated!! Best wishes, Chris Dr Christopher Johnson Department of Physiology, Medical Biology Centre, Queen's University of Belfast, 97 Lisburn Rd Belfast BT7 9BL 02890 272092 From gu.lang <@t> gmx.at Tue Nov 23 06:24:11 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Whole mount prostate specimens References: Message-ID: <003501c4d157$56919d40$eeeea8c0@server> Hi Angie, In our lab we do not perform whole mounting, but use the macro-cassettes and slides, that are 4 times larger than the standard ones. The doctor cuts the protstata in mostly 6 slices, after fixation for at least two days. Tissue processing requires double times than normal. We cut them on routine sliding microtomes (Microm) with a special blockholder. The cutting is not difficult. hope this helps Gudrun Akh Linz, Austria ----- Original Message ----- From: "Angela Bitting" To: Sent: Monday, November 22, 2004 8:53 PM Subject: [Histonet] Whole mount prostate specimens > I have been asked to research what type microtome, slides, coverslips, > knives I will need to begin cutting whole mount prostates. > If anyone out there is doing this in the clinical hospital setting, > could you help me out. Any procedures, hints, resources,....anything > will help. > What I'd really like to do is see someone actually cutting these type > specimens. Get some one-on-one training. I'm in central Pennsylvania if > anyone in the area wants to offer me some personal instruction. > Thanks folks! > Angie > > 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 Kemlo.Rogerson <@t> elht.nhs.uk Tue Nov 23 06:48:30 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] T2000 Non-gynae[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B516@bhrv-nt-11.bhrv.nwest.nhs.uk> We have a T2000 at East Lancs Cell Path and the young BMS in there is having problems. If my memory serves me there were threads in the past concerning Non-Gynae and the T2000. Her problem is that using the property cell transportation fluid, Cytolyt, she (and her Consultants), maintain cells are lost. To overcome that she is using the 'Pap TPAP' fluid that is used for cervical specimens but has difficulty lysing the rbc's; I suppose as they are fixed. I used to be a good 'technician' and I wonder if anyone could tell me how they receive FNAC, fluids, etc. I hope to revisit being a good 'technician' again, but frailty of my cerebral cortex prevents intuitive thought, so plagiarism is the only option. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals From michele.french <@t> bms.com Tue Nov 23 07:02:24 2004 From: michele.french <@t> bms.com (Michele French) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] IHC on Sections Processed for EM? Message-ID: <41A334E0.2060308@bms.com> Good Morning Histonet! A colleague of mine did some EM work and found something interesting. Our pathologist wanted me to try to do some IHC to further characterize what is present in the section. Our EM person said it would never work. I did not think it was possible either, but I thought I would ask anyway. Has anyone tried doing an immunostain on plastic (Epon) sections from tissue fixed and processed for EM? I am always up for a challenge, but I am really busy right now and don't want to waste my time if there is really no hope! Thanks in advance, Michele French From portera203 <@t> yahoo.com Tue Nov 23 08:33:25 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Elution reagent for Double Staining Message-ID: <20041123143325.59563.qmail@web40908.mail.yahoo.com> Anyone out there willing to share what they use for elution reagent during double immunostaining using peroxidase and phosphatase systems together? I have looked and gotten a variety of answers, I would like to know what actually works for people. Thanks in advance - 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!? Meet the all-new My Yahoo! – Try it today! From lloyd.3 <@t> osu.edu Tue Nov 23 09:21:24 2004 From: lloyd.3 <@t> osu.edu (Mary Lloyd) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] mucicarmine Message-ID: <5.1.0.14.2.20041123101518.00a86b10@pop.service.ohio-state.edu> I am having a problem with my mucicarmine stains. I always use Polyscientific kits but the last kit I received barely stained at all. I ordered mucicarmine from Fisher (Accustain) but it didn't work either. I am using salivary gland for the control so it should light up really well. Any suggestions to where I can either find a good mucin stain or technical hints. Thanks From gcallis <@t> montana.edu Tue Nov 23 09:38:59 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Elution reagent for Double Staining In-Reply-To: <20041123143325.59563.qmail@web40908.mail.yahoo.com> References: <20041123143325.59563.qmail@web40908.mail.yahoo.com> Message-ID: <6.0.0.22.1.20041123083306.01b407d0@gemini.msu.montana.edu> Talk to Chris van der Loos - he has done this and actually feels this does NOT work very well. There are other ways to get excellent results with double IHC and not use elution. He can be reached at "C.M. vander Loos" . There are problems with elution. One is the antibody is bound so strongly to antigen, it will not elute. Another is damage to antigens with elution reagents. Chris is a master at double staining and taught this in Toronto this past year - look for him to do this again in Fort Lauderdale, NSH symposium 2005 - he did discuss elution and how to get around it. A good visit with him may help you with your double staining. At 07:33 AM 11/23/2004, you wrote: >Anyone out there willing to share what they use for elution reagent during >double immunostaining using peroxidase and phosphatase systems >together? I have looked and gotten a variety of answers, I would like to >know what actually works for people. Thanks in advance - > > >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!? > Meet the all-new My Yahoo! ? Try it today! >_______________________________________________ >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 mcauliff <@t> umdnj.edu Tue Nov 23 13:11:25 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] IHC on Sections Processed for EM? In-Reply-To: <41A334E0.2060308@bms.com> References: <41A334E0.2060308@bms.com> Message-ID: <41A38B5D.6030505@umdnj.edu> Hi Michael: As usual, it "depends on the antigen". There are a few antigens that survive glut and osmium, I know that GFAP can be made to work on some astrocytes if the osmium is removed. Other than that, I would think a literature search for the antigen in question is the best first step. Maybe someone has made it work? Of course, the Epon will have to be etched. Geoff Michele French wrote: > Good Morning Histonet! A colleague of mine did some EM work and found > something interesting. Our pathologist wanted me to try to do some IHC > to further characterize what is present in the section. Our EM person > said it would never work. I did not think it was possible either, but > I thought I would ask anyway. Has anyone tried doing an immunostain on > plastic (Epon) sections from tissue fixed and processed for EM? I am > always up for a challenge, but I am really busy right now and don't > want to waste my time if there is really no hope! Thanks in advance, > Michele French > > _______________________________________________ > 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 slarkham <@t> qltinc.com Tue Nov 23 12:50:19 2004 From: slarkham <@t> qltinc.com (slarkham@qltinc.com) Date: Fri Sep 16 15:24:19 2005 Subject: Marking ink for maintaining orientation RE: [Histonet] freezing mouse eyes Message-ID: Yup i am with you on the marking with ink Gail, it works extremely well for us and then orientation isnt an issue, we use a dissecting microscope to put the ink on and then freeze using neg 50 (better than OCT in my eyes...heh no pun intended) in a cryomould and freeze on the surface of liquid nitrogen From michele.french <@t> bms.com Tue Nov 23 14:33:40 2004 From: michele.french <@t> bms.com (Michele French) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] IHC on EM Sections... Message-ID: <41A39EA4.5080009@bms.com> Thank you to everyone for all your comments and advice about doing IHC on sections from tissues procesed for EM. It really sounds challenging yet would be fun to try! I am going to compile all your thoughts as well as my own comments and forward them to my pathologist. Only he can decide whether the information gained would be worth the time and effort spent. If I do pursue the project, I will be happy to share the outcome at a later date. Michele From hymclab <@t> hyhc.com Tue Nov 23 15:23:16 2004 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] mucicarmine Message-ID: I recently had that same problem. Only at first we had Newcomer mucicarmine and then Poly Scientific. What finally solved it was making new Weigert's Hematoxylin. Someone must have made it up with too much HCl in solution B. It didn't occur to me that it was the Weigert's, I just thought it was the muci soltuion. When I made up fresh Weigert's being very careful on my measurements it turned out beautifully once again. When I looked back at the ones that didn't work I then noticed that the Weigert's did indeed look pale. I guess the extra acid in the Weigert's blocks it from picking up the mucicarmine solution. Hope this helps, I know how frustrating it is when a simple basic stain doesn't work. Also, the technical person at Newcomer had me try to de-gas my tap water by pouring some in a beaker and letting it stand overnight before I used it. I guess some water treatments facitlities sometimes puts too much chemical in and de-gassing helps, but that wasn't our problem. Dawn -----Original Message----- From: Mary Lloyd [mailto:lloyd.3@osu.edu] Sent: Tuesday, November 23, 2004 9:21 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] mucicarmine I am having a problem with my mucicarmine stains. I always use Polyscientific kits but the last kit I received barely stained at all. I ordered mucicarmine from Fisher (Accustain) but it didn't work either. I am using salivary gland for the control so it should light up really well. Any suggestions to where I can either find a good mucin stain or technical hints. Thanks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kwuny <@t> email.cs.nsw.gov.au Tue Nov 23 19:40:55 2004 From: kwuny <@t> email.cs.nsw.gov.au (Young Kwun) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] catecholamine/glyoxylic acid fluoresence In-Reply-To: <000001c4d156$6f7fb6d0$b53f758f@cjohnson> Message-ID: <200411241239604.SM01440@crgcsls814> Dear Dr Chris, Your fluorescence microscope setting seems OK. I used Olympus BH2 equipped with epi-illumination, and violet excitation and a Y-475 barrier filter. According to my old thesis, I used 2% glyoxylic acid and 15% sucrose in 0.1M phosphate buffer, pH adjusted to 5.0 with NaOH for 12 minutes. Magnesium chloride (0.5%) was added to the solution, immediately before use. After drying the slides I put into a gassing chamber for 5 min. Best wishes, Young Young Kwun, PhD Senior Hospital Scientist Dept of Anatomical Pathology Concord Hospital Concord NSW 2139 Australia Phone:61-2-9767 6075 Fax:61-2-9767 8427 Email:kwuny@email.cs.nsw.gov.au -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Chris Johnson Sent: Tuesday, 23 November 2004 11:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] catecholamine/glyoxylic acid fluoresence Dear All, I have recently been trying to show up sympathetic innervation of blood vessels (noradrenaline/norepinephrine. I have used rat tail artery (i.e., good sympathetic innervation) incubated in phosphate buffer and 2 % glyoxylic acid pH?d to 7.4 for 90 minutes followed 4 minutes at 100 ?C. At best I have seen very faint fluorescence with the very slightest hint of a nerve plexus. I don?t think it is the processing as I have used it successfully before and it seems to be fairly robust. I suspect it is our filtering, but on looking at the literature, I?m not sure. I am using Excitation 380 ? 400 nm Dichromatic 440 nm high pass Emission 455 ? 485 nm What are the emission characteristics of the fluorophore from this reaction? What am I doing wrong?!!!! Any suggestions would be greatly appreciated!! Best wishes, Chris Dr Christopher Johnson Department of Physiology, Medical Biology Centre, Queen's University of Belfast, 97 Lisburn Rd Belfast BT7 9BL 02890 272092 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please destroy it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of the Central Sydney Area Health Service." From moss <@t> cut.net Tue Nov 23 21:25:53 2004 From: moss <@t> cut.net (MOSS) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] A New Life -- Anybody want my job?? Message-ID: Hi everyone! This is a very exciting time for us in the McManus family. After 2 years of hard work, my husband?s business is finally ready to open our doors for business in January. We are doing electron microscopy, TEM to start out with and SEM will come on board when we are able to acquire the scope. I handed in my notice that I?m leaving in January, so an excellent job with great benefits and an awesome environment, cool boss and fun co-workers is open and up for grabs to anyone who qualifies. Contact the Employement office at Utah State University (www.usu.edu) and my boss, Dr. Tom Baldwin (ask me privately for his e-mail). Our new company is Mt Ogden Scientific Services (MOSS) and we are located in Ogden UT. We have an innovative method of getting final images out -- from fresh tissue to fixed, processed -- within 24 -72 hours. If there are any pathologists who are interested in our services, or if there are any EM labs that have more work than they can handle, please contact us. In the mean-time, have a great Thanksgiving everyone. Y?all outside the US, have a great week !! Connie McManus Mt Ogden Scientific Services Ogden UT 84401 Tel: 801/334-6677 website is forthcoming. From c.m.vanderloos <@t> amc.uva.nl Wed Nov 24 02:07:23 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] RE: how to identify what cell type is caspase3+ Message-ID: <84f5e87d38.87d3884f5e@amc.uva.nl> Hi Patsy, Like Luis already answered, double staining is the best way to identify caspase-3 positive cells. I have done this recently and indeed it worked very well. You are lucky in the first place that caspase-3 is a rabbit antibody that is easily combined with any mouse primary. Whether or not the cellular make-up during apoptosis is changed you never know before. This is just a matter of testing. I can assure you from my own experiments that double staining of caspase-3 combined with CD31 or SMA works for sure. Since caspase-3 shows both nuclear and cytoplasmic staining I stained CD31 or SMA with beta-galactosidase in turquoise and caspase-3 with alk. phosphatase in red (DakoCyto Liquid Permanent Red). If caspase shows up in the cytoplasm of a b-gal positive cell, co- localization is observed (blue/purple mixed-color). Optional is a very faint hematoxilin counterstain (use 1/10 diluted hematoxilin only for 15 sec or so) or Nuclear Red (15 sec). A very colorful experiment I can tell you! Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Patsy Ruegg Date Mon, 22 Nov 2004 11:40:54 -0700 To 'Histonet' Cc IHCRG@neo.agsci.colostate.edu, jenrueyt@hotmail.com Subject [Histonet] how to identify what cell type is caspase3+ I have a question from an investigator who is looking at rat lungs for apoptosis by using cleaved caspase 3 IHC and then wants to know what cell type is under going apoptosis (are they endothelial cells or a mix of different cells? how can they demonstrate the cell type that is positive for CC3 since the morphology changes when this happens) they do not appear to be smooth muscle cells since there is no positivity around the arteries. Should be be doing double IHC labeling say CC3 and CD31 or F8 together in the same section? Will the cell undergoing apoptosis still be expressing these other markers? Patsy ----- Original Message ----- >From Luis Chiriboga Date Mon, 22 Nov 2004 14:08:20 -0500 To Patsy Ruegg , 'Histonet' Cc IHCRG@neo.agsci.colostate.edu, jenrueyt@hotmail.com Subject [Histonet] RE: [IHCRG] how to identify what cell type is caspase3+ Hi Patsy Yes, That's exactly the approach we use.....double labeling should catch all CC3/Surface Marker+ cells except for those that have lost significant fractions of their cell membrane. Luis From naveedafahim <@t> hotmail.com Wed Nov 24 02:37:04 2004 From: naveedafahim <@t> hotmail.com (naveeda arshad) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] (no subject) Message-ID: _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From Kemlo.Rogerson <@t> elht.nhs.uk Wed Nov 24 02:38:22 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] A New Life -- Anybody want my job??[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B51F@bhrv-nt-11.bhrv.nwest.nhs.uk> Our thanks to our sponsors, MOSS Inc. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: MOSS [mailto:moss@cut.net] Sent: 24 November 2004 03:26 To: HISTONET Subject: [Histonet] A New Life -- Anybody want my job??[Scanned] Hi everyone! This is a very exciting time for us in the McManus family. After 2 years of hard work, my husband?s business is finally ready to open our doors for business in January. We are doing electron microscopy, TEM to start out with and SEM will come on board when we are able to acquire the scope. I handed in my notice that I?m leaving in January, so an excellent job with great benefits and an awesome environment, cool boss and fun co-workers is open and up for grabs to anyone who qualifies. Contact the Employement office at Utah State University (www.usu.edu) and my boss, Dr. Tom Baldwin (ask me privately for his e-mail). Our new company is Mt Ogden Scientific Services (MOSS) and we are located in Ogden UT. We have an innovative method of getting final images out -- from fresh tissue to fixed, processed -- within 24 -72 hours. If there are any pathologists who are interested in our services, or if there are any EM labs that have more work than they can handle, please contact us. In the mean-time, have a great Thanksgiving everyone. Y?all outside the US, have a great week !! Connie McManus Mt Ogden Scientific Services Ogden UT 84401 Tel: 801/334-6677 website is forthcoming. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bettina.hutz <@t> orionpharma.com Wed Nov 24 04:01:20 2004 From: bettina.hutz <@t> orionpharma.com (bettina.hutz@orionpharma.com) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] PSR Message-ID: <243E1A79AF5D89438C70513DBADAA4F0019E3FC9@sfies-exchange1.orionnet.org> ello, we have some difficulties with PSR staining and so far no trouble = shooting advice helped us finding the reason for our problem. We use PSR as routine staining in CV reserch studies in order to = measure the quantitive amount of connective tissue in heart slides. Therefore the staining has to be proper with a big contrast between yellow and red = areas. In the beginning all went well, but now the staining quality has = decreased, the connective trissue is not as bright stained as for example one year = ago. I use the same batch of picric acid and Pirco Siriusred and have not = changed anything in the staining procedure.=20 Does anyone has any idea what can be the reason? My staining instruction here: PSR (Picro-Sirius Red) staining of paraffin slides (modified by Bettina Hutz and Ritva Inkinen) 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 Weigert=B4s Hematoxylin 10 minutes Running tap water 10 minutes Rinse with aqua-dest. Picro-Sirius Red 30-60 minutes Dehydration & mounting 3x ethanol 100% short 2x xylene short Mount with xylene based medium 2. Solutions 2.1. Weigert=B4s Hematoxylin 2.1.1 Stock solution A Hematoxylin (Natural Black 1, C.I. 75290) 10,0 g Ethanol 95% 1000 ml Produce app. 2 weeks before use 2.1.2. Stock solution B Ferric chloride, 29% aqueous 40 ml Aqua-dest. 950 ml HCl conc. 10 ml 2.1.3 Usage solution Equal parts of solution A and solution B, use immediately. 2.2 Picro-Sirius Red 2.2.1 Stock solution Sirius Red F3BA 5,0 g Picric acid, saturated, aqueous 500 ml Allow to stand for 48 hours before producing the usage solution. Thanks for your help:) Bettina Hutz 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 From c.m.vanderloos <@t> amc.uva.nl Wed Nov 24 04:12:40 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] RE: Elution reagent for Double Staining Message-ID: Dear Amy, Gayle is right: I don't regard sequential double staining with elution in between as a very safe way of double staining. Only in case you wish to stain to different cell populations, and you are sure there is no co- localization involved, you may try sequential double staining. The DakoCyto EnVision Doublestain kit is worthwhile to test. The so called "doublestain block" is equal to the low pH glycin-HCl buffer and isn't working either. Some remarks: 1. Sequential double staining only works because of unique effective sheltering of DAB chromogen. As far as I know there are no other enzymatic chromogens with this sheltering capacity. Always apply DAB after the 1st staining sequence!!!!! 2. You may try to remove reagents of the 1st staining sequence using glycing-HCl buffer (0.1 M, pH2.2) however high affinity primaries will stay at your tissue section anyway. In the DakoCyto EnVision Doublestain kit the "doublestain block" is equal to the low pH glycin- HCl buffer and isn't working either for removing high affinity primaries. Always test your double staining with/without the elution step. 3. Glycin-HCl buffer may damage the tissue morphology of an acetone- fixed cryostat section. Don't do this. 4. Again: sequential double staining technique is not for visualizing co-localization by mixed-colors! Furthermore, the brown-red (or brown- blue if you wish) color combi is very poor for visualizing a distinguishable mixed-color. Don't do this. 5. If you are immunostaining two antigens that are at very closely together you may end up in trouble: DAB after the 1st staining sequence will effectively shelter the 2nd antigen too. Hope these do-s and don't-s work for you. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Gayle Callis Date Tue, 23 Nov 2004 08:38:59 -0700 To Amy Porter , Histonet@lists.utsouthwestern.edu Subject Re: [Histonet] Elution reagent for Double Staining Talk to Chris van der Loos - he has done this and actually feels this does NOT work very well. There are other ways to get excellent results with double IHC and not use elution. He can be reached at "C.M. vander Loos" . There are problems with elution. One is the antibody is bound so strongly to antigen, it will not elute. Another is damage to antigens with elution reagents. Chris is a master at double staining and taught this in Toronto this past year - look for him to do this again in Fort Lauderdale, NSH symposium 2005 - he did discuss elution and how to get around it. A good visit with him may help you with your double staining. At 07:33 AM 11/23/2004, you wrote: >Anyone out there willing to share what they use for elution reagent during >double immunostaining using peroxidase and phosphatase systems >together? I have looked and gotten a variety of answers, I would like to >know what actually works for people. Thanks in advance - > >Amy S.Porter, HT(ASCP) >Michigan State University >Department of Physiology >Division of Human Pathology >College of Human Medicine >portera203@yahoo.com From yej2 <@t> leicester.ac.uk Wed Nov 24 06:13:49 2004 From: yej2 <@t> leicester.ac.uk (El Jai, Y.) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Beta gal staining. Message-ID: <1F2CE8D4B0195E488213E8B8CCF71486034C96C1@Saffron.cfs.le.ac.uk> I have a mouse expressing a fusion between a protein of interest and beta-gal. I want to look at expression in different embryonic stages and tissues. I have tried cryosections followed by x-gal staining but the morphology is not great. Has anyone got a protocol to do the staining and the sectioning in a different way. Thanks for your help Yasmine. PS: thanks for transmitting this message to the board. ---------------------------- Yasmine El Jai Lab 205 Dept of Biochemistry University Road University of Leicester UK +44.1162523475 ---------------------------- From jkiernan <@t> uwo.ca Sat Nov 20 00:18:25 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] Re: Methods Mol Med Publication is out References: Message-ID: <419EE1B1.82448FA@uwo.ca> Dear G. Steven Bova, The PubMed link in your email shows me as a co-author of a multi-author chapter. I never consented to any such co-authorship. Why have you done this? Who is your publisher? I am not one of your 7 "coauthors". Your email mentions Humana Press and NIH. What are you doing? You could be digging yourself deeply into all sorts of legal troubles. - ------------------------------- 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 _______________________________ "G. Steven Bova" wrote: > > Dear Isam, John, Gene, Andra, Carolyn, John, and Mike: > > I just wanted you to know that the book-version of our chapter has been > published: > > http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15542897 > > Thanks so much for all your help on it. It was arduous to put > together. It has a lot of good basic information in it that I think > people will find helpful. > > Snce this book is targeted mainly to the Pancreatic Cancer research > audience, Humana agreed to publish it also in a reformatted version in > the journal Molecular Biotechnology---but I haven't been given a > specific date of publication. > > Have a good weekend, > > Steve > > G. Steven Bova M.D. > Asst Prof. of Pathology, Health Sci Informatics, Genetic Medicine, > Oncology, Urology > Department of Pathology > Johns Hopkins University School of Medicine > Carnegie Building Rm 628 > 600 North Wolfe St. > Baltimore, MD 21287-6417 > > Email: gbov@jhmi.edu > Fax: 410-614-7620 > Voicemail: 410-614-5957 ------------------------------------------- From GHAWKINS <@t> NORTHERNHEALTH.ORG Wed Nov 24 09:08:02 2004 From: GHAWKINS <@t> NORTHERNHEALTH.ORG (Gerald Hawkins) Date: Fri Sep 16 15:24:19 2005 Subject: [Histonet] TISSUE TEK VIP Message-ID: I know that there is a wide range ambient temp for operating the instrument, however, is there preferred optimal range and how can it be expected to be maintained reasonably??What is the temp that each station should be expected to be at??? Thank you for your time...Jerry From jkiernan <@t> uwo.ca Wed Nov 24 10:15:50 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] PSR References: <243E1A79AF5D89438C70513DBADAA4F0019E3FC9@sfies-exchange1.orionnet.org> Message-ID: <41A4B3B6.46E6E7F3@uwo.ca> Two details are not clear from your method. 1. Does the picro-sirius red solution contain some undissolved picric acid (deposit in bottom of bottle)? It should, to ensure saturation. 2. Is there an intervening wash between dyeing in the picro-sirius solution and the first of the 3 changes of 100% alcohol? You should not wash in water, but slightly acidified water (eg approx 0.1% acetic acid) is desirable to rinse away only unbound dyes. Water that is not acidified will remove some sirius red, and you may lose some finer fibres, basement membranes etc. As you have probably noticed, nuclear staining by Weigert's iron-haematoxylin is weakened by subsequently applied picro-sirius red (in contrast to Van Gieson's picro-acid fuchsine). This is because of the longer time in an acidic medium (30 min in contrast to less than 5 with van G). For your purposes the nuclear stain seems pointless. With your rapid dehydration you are conserving the yellow cytoplasmic (muscle) stain, so you should be able to measure the total area and the red (collagen) area. Picric acid is rather easily removed by alcohol; most users of the picro-sirius red method do not mind this, or even prefer an unstained cytoplasmic background. If you are using picro-sirius red for research purposes you should be familiar with the classic papers of Puchtler and of Junqueira from the 1960s and 1970s. A Google search for Puchtler and Junqueira brings up some 25 items; several are Histonet replies with technical instructions and lists of references to get you started in the library. -- ------------------------------- 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 _______________________________ bettina.hutz@orionpharma.com wrote: > > ello, > > we have some difficulties with PSR staining and so far no trouble = shooting > advice helped us finding the reason for our problem. We use PSR as routine > staining in CV reserch studies in order to = measure the quantitive amount > of connective tissue in heart slides. Therefore the staining has to be > proper with a big contrast between yellow and red = areas. In the beginning > all went well, but now the staining quality has = decreased, the connective > trissue is not as bright stained as for example one year = ago. I use the > same batch of picric acid and Pirco Siriusred and have not = changed > anything in the staining procedure.=20 Does anyone has any idea what can be > the reason? My staining instruction here: > > PSR (Picro-Sirius Red) staining of paraffin slides > (modified by Bettina Hutz and Ritva Inkinen) > 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 > Weigert=B4s Hematoxylin 10 minutes > Running tap water 10 minutes > Rinse with aqua-dest. > Picro-Sirius Red 30-60 minutes > Dehydration & mounting > 3x ethanol 100% short > 2x xylene short > Mount with xylene based medium > 2. Solutions > 2.1. Weigert=B4s Hematoxylin > > 2.1.1 Stock solution A > Hematoxylin (Natural Black 1, C.I. 75290) 10,0 g > Ethanol 95% 1000 ml > Produce app. 2 weeks before use > > 2.1.2. Stock solution B > Ferric chloride, 29% aqueous 40 ml > Aqua-dest. 950 ml > HCl conc. 10 ml > > 2.1.3 Usage solution > Equal parts of solution A and solution B, use immediately. > 2.2 Picro-Sirius Red > > 2.2.1 Stock solution > Sirius Red F3BA 5,0 g > Picric acid, saturated, aqueous 500 ml > Allow to stand for 48 hours before producing the usage solution. > > Thanks for your help:) > > Bettina Hutz > > 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 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BriggsK <@t> drmc.drhsi.org Wed Nov 24 11:13:33 2004 From: BriggsK <@t> drmc.drhsi.org (Kevin Briggs) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Cryostat Repair Message-ID: <7C1D5BE1ADF06E488AEB7A6247214A860D250D@HORNET.drmc.drhsi.org> Hello, does anyone know of a good repair center or person in Virginia or North Carolina who services cryostats? Thanks, Kevin D. Briggs, MS, CT(ASCP) Team Leader- Cytopathology/ Histopathology Services Danville Regional Medical Center 142 South Main Street Danville, VA 24541 Telephone: (434) 799-4470 ext.5451 Fax: (434) 799-2118 E-mail: briggsk@drhsi.org From mlgiebel <@t> mail2.vcu.edu Wed Nov 24 12:02:14 2004 From: mlgiebel <@t> mail2.vcu.edu (Mary Lee Giebel) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Cryostat Repair In-Reply-To: <7C1D5BE1ADF06E488AEB7A6247214A860D250D@HORNET.drmc.drhsi.org> Message-ID: <200411241802.NAA24075@arrakis.vcu.edu> Kevin, The company that we have used is: Southern Air, Inc. 2655 Lakeside Drive Lynchburg, VA 24501 (804) 285-6528 They have offices all over the state. I have also used another company, H&M Sales and Service (804)276-4668, but they do not cover the Danville area. We have been very pleased with both companies. Good Luck with your repair. Regards, Mary Lee Giebel Manager and HTL Massey Cancer Center Research Histopathology Core Virginia Commonwealth University Sanger Hall Room 4-036 1101 East Marshall Street Richmond, Virginia 23298 phone (804) 828-5092 fax (804) 828-9749 ------------------- > Hello, > > does anyone know of a good repair center or person in Virginia or North Carolina who services cryostats? > > Thanks, > > Kevin D. Briggs, MS, CT(ASCP) > Team Leader- Cytopathology/ Histopathology Services > Danville Regional Medical Center > 142 South Main Street > Danville, VA 24541 > Telephone: (434) 799-4470 ext.5451 > Fax: (434) 799-2118 > E-mail: briggsk@drhsi.org > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From eearle <@t> abrazohealth.com Wed Nov 24 12:13:32 2004 From: eearle <@t> abrazohealth.com (Earle, Elizabeth) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Voice recognition software Message-ID: <4727B24D9A8575479222635B57E3B3DB8CF728@mail.vhswest.local> To all who are using voice recognition, some help, please: Which system do you use? How long have you used it? What do you like/not like about it? If any vendors want to respond to me directly, that is fine Thanks E Earle From DNolan <@t> evanhospital.com Wed Nov 24 12:12:59 2004 From: DNolan <@t> evanhospital.com (Donna M. Nolan) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] CBG recyler Message-ID: <214BD65A9D60934BA8EA577AE5AD280928ECB7@EVANXSCL.evanhospital.net> We have a CBG solvent recycler that we are using for Xylene, Shandon Xylene Substitue and ethanol alcohols. The problem is the alcohol is contaminated with the solvent. We did a cleaning and the alcohol was fine but the first time we ran Xylene through, did the flush and ran alcohol the alcohol was once again contaminated. Anyone have this happen? Does anyone else try to run solvents and alcohol on the same recycler? CBG says we can and they have been helpful in trying to solve this but so far no luck. Donna Nolan HT(ASCP) Evangelical Community Hospital From DNolan <@t> evanhospital.com Wed Nov 24 12:17:11 2004 From: DNolan <@t> evanhospital.com (Donna M. Nolan) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] processor - alcohol contamination Message-ID: <214BD65A9D60934BA8EA577AE5AD280928ECB8@EVANXSCL.evanhospital.net> The absolute alcohol stations on our processor have been getting contaminated with our clearing reagent. The processor is approx. 9 years old. We have had service out but they have not been able to find the problem. We probably would not have noticed if we didn't recycle our alcohols and started getting contaminated alcohol which we traced back to the processor. I also have a post about our recycler and contamination. I would appreciate input on these problems. Donna Nolan HT(ASCP) Evangelical Community Hospital. From JQB7 <@t> CDC.GOV Wed Nov 24 12:27:00 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] processor - alcohol contamination Message-ID: What brand/model of processor? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Donna M. Nolan Sent: Wed 11/24/2004 1:17 PM To: Histonet Cc: Subject: [Histonet] processor - alcohol contamination The absolute alcohol stations on our processor have been getting contaminated with our clearing reagent. The processor is approx. 9 years old. We have had service out but they have not been able to find the problem. We probably would not have noticed if we didn't recycle our alcohols and started getting contaminated alcohol which we traced back to the processor. I also have a post about our recycler and contamination. I would appreciate input on these problems. Donna Nolan HT(ASCP) Evangelical Community Hospital. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From scoop <@t> mail.nih.gov Wed Nov 24 12:40:40 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] F4/80 on formalin fixed frozen spleen Message-ID: Does anyone know if F4/80 can work on formalin fixed frozen mouse spleen for IHC without antigen retrieval? Also, thanks to everyone for their suggestions re. mouse B-cell markers. Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From gcallis <@t> montana.edu Wed Nov 24 12:50:52 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] F4/80 on formalin fixed frozen spleen In-Reply-To: References: Message-ID: <6.0.0.22.1.20041124114859.01b34470@gemini.msu.montana.edu> If you have to do retrieval on frozen section fixed with formalin, then you may lose section. Why do just do paraffin, and then retrieval for F4/80. Other labs have gotten this to work, and has been posted on Histonet - check out archives for information. At 11:40 AM 11/24/2004, you wrote: >Does anyone know if F4/80 can work on formalin fixed frozen mouse spleen >for IHC without antigen retrieval? > >Also, thanks to everyone for their suggestions re. mouse B-cell markers. > >Thanks, >Sharon >-- >Sharon Cooperman >NIH, NICHD, CBMB 301.435-8417 >Building 18T, room 101 301.402-0078 fax >Bethesda, MD 20892 > >_______________________________________________ >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 komuves <@t> corgentech.com Wed Nov 24 13:44:28 2004 From: komuves <@t> corgentech.com (Laszlo Komuves) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] RE: IHC on tissues processed for EM Message-ID: There are hundreds, if not thousands of papers demonstrating successful localization of intracellular antigens at EM or LM level using tissue samples processed conventionally for EM (i.e. PFA/GA fixation, Os postfixation, epoxy resin embedding). Look for the now classical papers by Roth and Bendayan or their early reviews. Also the EM textbooks published in the 80's have plenty of information. And let me also add a personal comment of frustration: Histonet used to be about sharing experience and wisdom, mentoring and teaching those have were (are they still?) eager to learn or perfect their skills. I personally learnt/benefited from many messages posted. Now people are asking advice about coverslip sizes? So thanks to the curse of the Internet, journal articles and text books are used only by a dedicated few, electronic databases (libraries, PubMed, journal archives, reagent search sites, even Google) are not searched, and now even the vendor catalogs are not opened, because on the Histonet somebody will respond? How will users differentiate between solid information (based on scientific knowledge, practical experience) or just plain ignorance? Where is critical thinking? What happened to a once honorable guild of skilled craftsmanship, when members of the community are lacking elementary knowledge (just a few recent hair-raising topics: H&E staining, pH, and the list goes on and on)? And of course we/they are constantly offended/complain about the lack of recognition by our/their peers. Feel free to get angry with me. I certainly welcome arguments and criticism.. As for me, I go back to find something useful in one of my textbooks. Laszlo Komuves FROM: L?szl? G. Komuves PhD Senior Principal Scientist, Manager, Microscopy Core Laboratory CORGENTECH, Inc. 650 Gateway Blvd., South San Francisco, CA 94080 Phone: (650) 246-6905, Fax: (650) 624-7540 E-mail: komuves@corgentech.com Date: Tue, 23 Nov 2004 08:02:24 -0500 From: Michele French Subject: [Histonet] IHC on Sections Processed for EM? To: histonet@lists.utsouthwestern.edu Message-ID: <41A334E0.2060308@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Good Morning Histonet! A colleague of mine did some EM work and found something interesting. Our pathologist wanted me to try to do some IHC to further characterize what is present in the section. Our EM person said it would never work. I did not think it was possible either, but I thought I would ask anyway. Has anyone tried doing an immunostain on plastic (Epon) sections from tissue fixed and processed for EM? I am always up for a challenge, but I am really busy right now and don't want to waste my time if there is really no hope! Thanks in advance, Michele French From victor <@t> pathology.washington.edu Wed Nov 24 13:48:26 2004 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] A New Life -- Anybody want my job?? In-Reply-To: References: Message-ID: <41A4E58A.6060500@pathology.washington.edu> I can vouch that her boss is coooooooooool. I worked with Dr. Baldwin back when he was a part of Washington Animal Disease Diagnostic Laboratory. Brings back some fond memories. Say hi to Tom for me. In case he forgets who I am, I supervised the Histology lab. Victor MOSS wrote: >Hi everyone! > >This is a very exciting time for us in the McManus family. After 2 years of >hard work, my husband?s business is finally ready to open our doors for >business in January. We are doing electron microscopy, TEM to start out >with and SEM will come on board when we are able to acquire the scope. > > I handed in my notice that I?m leaving in January, so an excellent job with >great benefits and an awesome environment, cool boss and fun co-workers is >open and up for grabs to anyone who qualifies. Contact the Employement >office at Utah State University (www.usu.edu) and my boss, Dr. Tom Baldwin >(ask me privately for his e-mail). > >Our new company is Mt Ogden Scientific Services (MOSS) and we are located in >Ogden UT. We have an innovative method of getting final images out -- from >fresh tissue to fixed, processed -- within 24 -72 hours. If there are any >pathologists who are interested in our services, or if there are any EM labs >that have more work than they can handle, please contact us. > >In the mean-time, have a great Thanksgiving everyone. Y?all outside the US, >have a great week !! > > >Connie McManus >Mt Ogden Scientific Services >Ogden UT 84401 >Tel: 801/334-6677 >website is forthcoming. > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax From koellinr <@t> amgen.com Wed Nov 24 13:57:38 2004 From: koellinr <@t> amgen.com (Koelling, Ray) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] RE: IHC on tissues processed for EM Message-ID: <16834C6DFFA6004C88DE4507FB8AE544018CC306@wa-mb4-sea.amgen.com> Not only would I not get angry but more than happy to meet you and buy you a Seattle seafood meal for saying it if you ever venture up this way. And I'm not kidding. Raymond Koelling Resaerch Scientist, Pathology Amgen Corp. Seattle, WA 98119 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Laszlo Komuves Sent: Wednesday, November 24, 2004 11:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: IHC on tissues processed for EM There are hundreds, if not thousands of papers demonstrating successful localization of intracellular antigens at EM or LM level using tissue samples processed conventionally for EM (i.e. PFA/GA fixation, Os postfixation, epoxy resin embedding). Look for the now classical papers by Roth and Bendayan or their early reviews. Also the EM textbooks published in the 80's have plenty of information. And let me also add a personal comment of frustration: Histonet used to be about sharing experience and wisdom, mentoring and teaching those have were (are they still?) eager to learn or perfect their skills. I personally learnt/benefited from many messages posted. Now people are asking advice about coverslip sizes? So thanks to the curse of the Internet, journal articles and text books are used only by a dedicated few, electronic databases (libraries, PubMed, journal archives, reagent search sites, even Google) are not searched, and now even the vendor catalogs are not opened, because on the Histonet somebody will respond? How will users differentiate between solid information (based on scientific knowledge, practical experience) or just plain ignorance? Where is critical thinking? What happened to a once honorable guild of skilled craftsmanship, when members of the community are lacking elementary knowledge (just a few recent hair-raising topics: H&E staining, pH, and the list goes on and on)? And of course we/they are constantly offended/complain about the lack of recognition by our/their peers. Feel free to get angry with me. I certainly welcome arguments and criticism.. As for me, I go back to find something useful in one of my textbooks. Laszlo Komuves FROM: L?szl? G. Komuves PhD Senior Principal Scientist, Manager, Microscopy Core Laboratory CORGENTECH, Inc. 650 Gateway Blvd., South San Francisco, CA 94080 Phone: (650) 246-6905, Fax: (650) 624-7540 E-mail: komuves@corgentech.com Date: Tue, 23 Nov 2004 08:02:24 -0500 From: Michele French Subject: [Histonet] IHC on Sections Processed for EM? To: histonet@lists.utsouthwestern.edu Message-ID: <41A334E0.2060308@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Good Morning Histonet! A colleague of mine did some EM work and found something interesting. Our pathologist wanted me to try to do some IHC to further characterize what is present in the section. Our EM person said it would never work. I did not think it was possible either, but I thought I would ask anyway. Has anyone tried doing an immunostain on plastic (Epon) sections from tissue fixed and processed for EM? I am always up for a challenge, but I am really busy right now and don't want to waste my time if there is really no hope! Thanks in advance, Michele French _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Thu Nov 25 03:08:31 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] RE: IHC on tissues processed for EM References: Message-ID: <003a01c4d2ce$55afff70$eeeea8c0@server> Laszlo, I feel about saying some words about your message. You complain, that members of the histonetlist have no basic knowledgment. But what is basic? For me, in a routine surgery histolab, EM is very, very special for example. I belong to those people, that work in a small corner of the whole histological techniques. But I am interested and want to learn. You say, people have to use databases, text books etc. In my case I have no access through my lab to those things and they are'nt cheap (and first you have to know where to look). I am very happy about the histonetlist. Here are specialists, experts from every field and also "workmates". Please be tolerant with "easy" questions and problems, perhaps the person considers the problem more difficult than you know. friendly greetings Gudrun Lang ----- Original Message ----- From: "Laszlo Komuves" To: Sent: Wednesday, November 24, 2004 8:44 PM Subject: [Histonet] RE: IHC on tissues processed for EM There are hundreds, if not thousands of papers demonstrating successful localization of intracellular antigens at EM or LM level using tissue samples processed conventionally for EM (i.e. PFA/GA fixation, Os postfixation, epoxy resin embedding). Look for the now classical papers by Roth and Bendayan or their early reviews. Also the EM textbooks published in the 80's have plenty of information. And let me also add a personal comment of frustration: Histonet used to be about sharing experience and wisdom, mentoring and teaching those have were (are they still?) eager to learn or perfect their skills. I personally learnt/benefited from many messages posted. Now people are asking advice about coverslip sizes? So thanks to the curse of the Internet, journal articles and text books are used only by a dedicated few, electronic databases (libraries, PubMed, journal archives, reagent search sites, even Google) are not searched, and now even the vendor catalogs are not opened, because on the Histonet somebody will respond? How will users differentiate between solid information (based on scientific knowledge, practical experience) or just plain ignorance? Where is critical thinking? What happened to a once honorable guild of skilled craftsmanship, when members of the community are lacking elementary knowledge (just a few recent hair-raising topics: H&E staining, pH, and the list goes on and on)? And of course we/they are constantly offended/complain about the lack of recognition by our/their peers. Feel free to get angry with me. I certainly welcome arguments and criticism.. As for me, I go back to find something useful in one of my textbooks. Laszlo Komuves FROM: L?szl? G. Komuves PhD Senior Principal Scientist, Manager, Microscopy Core Laboratory CORGENTECH, Inc. 650 Gateway Blvd., South San Francisco, CA 94080 Phone: (650) 246-6905, Fax: (650) 624-7540 E-mail: komuves@corgentech.com Date: Tue, 23 Nov 2004 08:02:24 -0500 From: Michele French Subject: [Histonet] IHC on Sections Processed for EM? To: histonet@lists.utsouthwestern.edu Message-ID: <41A334E0.2060308@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Good Morning Histonet! A colleague of mine did some EM work and found something interesting. Our pathologist wanted me to try to do some IHC to further characterize what is present in the section. Our EM person said it would never work. I did not think it was possible either, but I thought I would ask anyway. Has anyone tried doing an immunostain on plastic (Epon) sections from tissue fixed and processed for EM? I am always up for a challenge, but I am really busy right now and don't want to waste my time if there is really no hope! Thanks in advance, Michele French _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mpool <@t> ohri.ca Thu Nov 25 13:15:08 2004 From: mpool <@t> ohri.ca (Pool, Madeline) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] blotches in sections Message-ID: <0133EAC2EDC7D74D944267D639E49EF203EBD6@ohexch04.civic1.ottawahospital.on.ca> I'm cutting 10 um sections from paraffin embedded mouse brain, spinal cord, and optic nerve and staining them with Luxol Fast Blue. In the brain sections, I'm seeing some splotchy darker background. I'm wondering if maybe I didn't dry the slides enough after sectioning or maybe heated it too much or something else entirely. If anyone has seen things like this before and can tell me what I should do to fix it, please let me know. I've put a picture of a section up at: http://members.rogers.com/madeline.pool/brain.jpg (Don't worry about the tearing, I was testing out bad sections.) Thanks in advance, Madeline Madeline Pool Ottawa Health Research Institute 501 Smyth Road Ottawa, Ontario K1H 8L6 (613)739-6634 fax: (613)737-8023 From AnthonyH <@t> chw.edu.au Thu Nov 25 15:49:33 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] blotches in sections Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E121@simba.kids> Madeline, My first impression is that there is water on the sections. You may possibly have to prolong your dehydration and clearing prior to coverslipping. You could decoverslip, remove the mountant in xylene and complete the dehydration in ethanol and then (via xylene or its substitute) recoverslip. 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: Pool, Madeline [mailto:mpool@ohri.ca] Sent: Friday, 26 November 2004 6:15 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] blotches in sections I'm cutting 10 um sections from paraffin embedded mouse brain, spinal cord, and optic nerve and staining them with Luxol Fast Blue. In the brain sections, I'm seeing some splotchy darker background. I'm wondering if maybe I didn't dry the slides enough after sectioning or maybe heated it too much or something else entirely. If anyone has seen things like this before and can tell me what I should do to fix it, please let me know. I've put a picture of a section up at: http://members.rogers.com/madeline.pool/brain.jpg (Don't worry about the tearing, I was testing out bad sections.) Thanks in advance, Madeline Madeline Pool Ottawa Health Research Institute 501 Smyth Road Ottawa, Ontario K1H 8L6 (613)739-6634 fax: (613)737-8023 _______________________________________________ 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 Kemlo.Rogerson <@t> elht.nhs.uk Fri Nov 26 02:25:45 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] blotches in sections[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B536@bhrv-nt-11.bhrv.nwest.nhs.uk> Seen it once with poor processing, there is an area around the artefact that appears ragged as if it didn't cut properly. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 From Kemlo.Rogerson <@t> elht.nhs.uk Fri Nov 26 02:27:01 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] blotches in sections[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B537@bhrv-nt-11.bhrv.nwest.nhs.uk> Ooops sorry, didn't read Tony's reply. I concur. 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: 25 November 2004 21:50 To: 'Pool, Madeline'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] blotches in sections[Scanned] Madeline, My first impression is that there is water on the sections. You may possibly have to prolong your dehydration and clearing prior to coverslipping. You could decoverslip, remove the mountant in xylene and complete the dehydration in ethanol and then (via xylene or its substitute) recoverslip. 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: Pool, Madeline [mailto:mpool@ohri.ca] Sent: Friday, 26 November 2004 6:15 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] blotches in sections I'm cutting 10 um sections from paraffin embedded mouse brain, spinal cord, and optic nerve and staining them with Luxol Fast Blue. In the brain sections, I'm seeing some splotchy darker background. I'm wondering if maybe I didn't dry the slides enough after sectioning or maybe heated it too much or something else entirely. If anyone has seen things like this before and can tell me what I should do to fix it, please let me know. I've put a picture of a section up at: http://members.rogers.com/madeline.pool/brain.jpg (Don't worry about the tearing, I was testing out bad sections.) Thanks in advance, Madeline Madeline Pool Ottawa Health Research Institute 501 Smyth Road Ottawa, Ontario K1H 8L6 (613)739-6634 fax: (613)737-8023 _______________________________________________ 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 corporate_21 <@t> hotmail.com Fri Nov 26 04:29:43 2004 From: corporate_21 <@t> hotmail.com (corporate headquarters) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Equipment from Biogenex, Vision Bio, Dako and Ventana In-Reply-To: Message-ID: Congratulations Connie ! Sounds like a wonderful time for you. My best wishes for success at your new Company. If you are in need of some instrumentation as you grow, I hear Vision Biosystems has a pretty good Tissue processor on the market. The Tissue Tek is a pretty good standard as well. If you are in need of other equipment, their are several excellent sources on the internet, which I'm sure you know. Is your job still available? If so, I would like to make contact and send a resume. I have 10 years in the IHC market, and am looking to make a move soon as our Company has filed for bankruptcy. If your Company in Utah is in the market for other Histology products, I would look into their other IHC systems, and get more info. Dako has had a good quarter, as has Biogenex. Ventana is for the BIG labs and Vison Biosystem is embroiled in litigation with Ventana and Dako. Re Ventana, It appears from the websites that the Court ruled that Vision infringed upon Ventana bar code patents, which prevents them from being able to distribute their Bond-X and MAX autostainers. Re-work could take up to a year, and they are not allowed by law to sell them while the Court ruling is in effect. A Court appeal may take less time. Dako is going after Vision's Director of Service and Support for actively trying to 'steal' Field Service Engineers from them, and it may be the case that the Director of Sales is passing on Confidential Dako info - just a rumor. I overheard at NSH Dako's Corporate strategy for 2003-2007 from people speaking at the Vision booth. Oh, well. This is an interesting business. Very best of luck in the new endeavors. Sincerely, Edgar Renton, HTL (ASCP) Mt. Sinai Medical Systems Chicago, IL >From: MOSS >To: HISTONET >Subject: [Histonet] A New Life -- Anybody want my job?? >Date: Tue, 23 Nov 2004 20:25:53 -0700 > >Hi everyone! > >This is a very exciting time for us in the McManus family. After 2 years >of >hard work, my husband¹s business is finally ready to open our doors for >business in January. We are doing electron microscopy, TEM to start out >with and SEM will come on board when we are able to acquire the scope. > > I handed in my notice that I¹m leaving in January, so an excellent job >with >great benefits and an awesome environment, cool boss and fun co-workers is >open and up for grabs to anyone who qualifies. Contact the Employement >office at Utah State University (www.usu.edu) and my boss, Dr. Tom Baldwin >(ask me privately for his e-mail). > >Our new company is Mt Ogden Scientific Services (MOSS) and we are located >in >Ogden UT. We have an innovative method of getting final images out -- from >fresh tissue to fixed, processed -- within 24 -72 hours. If there are >any >pathologists who are interested in our services, or if there are any EM >labs >that have more work than they can handle, please contact us. > >In the mean-time, have a great Thanksgiving everyone. Y¹all outside the >US, >have a great week !! > > >Connie McManus >Mt Ogden Scientific Services >Ogden UT 84401 >Tel: 801/334-6677 >website is forthcoming. > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From bruyntjes <@t> voeding.tno.nl Fri Nov 26 05:12:15 2004 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] CD23 and/or Fc epsilon receptor I alpha Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079D57@ntexch1.voeding.tno.nl> Hi all Is anyone of you aware of the cross-reactivity, or existence of an antibody that labels cd23 (low affinity IgE receptor) or FC epsilon receptor 1 alpha (high affinity IgE receptor) in rat tissue? Joost Bruijntjes TNO Nutrition and Food Research PO Box 360 3700 AJ Zeist The Netherlands This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From Andrew.Wong <@t> ed.ac.uk Fri Nov 26 07:55:05 2004 From: Andrew.Wong <@t> ed.ac.uk (Andrew Wong) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] F4/80 on formalin fixed frozen spleen In-Reply-To: <200411251809.iAPI9Quf021211@haymarket.ed.ac.uk> References: <200411251809.iAPI9Quf021211@haymarket.ed.ac.uk> Message-ID: <1101477305.41a735b986b03@sms.ed.ac.uk> I've had good results with F4/80 staining on formaldehyde fixed cryostat sections of mouse brain. However, it did require microwave/citric acid antigen retrieval. Trypsinisation pre-treatment also works, but I found it to be harsher on the sections than microwaving. Regards, Andrew -- Andrew Wong Division of Neuroscience University of Edinburgh 1 George Square Edinburgh EH8 9JZ Andrew.Wong@ed.ac.uk T: 0131 6511 875 > > Message: 6 > Date: Wed, 24 Nov 2004 13:40:40 -0500 > From: Sharon Cooperman > Subject: [Histonet] F4/80 on formalin fixed frozen spleen > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset="us-ascii" ; format="flowed" > > Does anyone know if F4/80 can work on formalin fixed frozen mouse > spleen for IHC without antigen retrieval? > > Also, thanks to everyone for their suggestions re. mouse B-cell > markers. > > Thanks, > Sharon > -- > Sharon Cooperman > NIH, NICHD, CBMB 301.435-8417 > Building 18T, room 101 301.402-0078 fax > Bethesda, MD 20892 > > > > From pruegg <@t> ihctech.net Fri Nov 26 08:54:47 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] F4/80 on formalin fixed frozen spleen In-Reply-To: <1101477305.41a735b986b03@sms.ed.ac.uk> Message-ID: I use this antibody on ffpe ms tissues using proteinase K digestion for 5 min. no HIER. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Andrew Wong Sent: Friday, November 26, 2004 6:55 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] F4/80 on formalin fixed frozen spleen I've had good results with F4/80 staining on formaldehyde fixed cryostat sections of mouse brain. However, it did require microwave/citric acid antigen retrieval. Trypsinisation pre-treatment also works, but I found it to be harsher on the sections than microwaving. Regards, Andrew -- Andrew Wong Division of Neuroscience University of Edinburgh 1 George Square Edinburgh EH8 9JZ Andrew.Wong@ed.ac.uk T: 0131 6511 875 > > Message: 6 > Date: Wed, 24 Nov 2004 13:40:40 -0500 > From: Sharon Cooperman > Subject: [Histonet] F4/80 on formalin fixed frozen spleen > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset="us-ascii" ; format="flowed" > > Does anyone know if F4/80 can work on formalin fixed frozen mouse > spleen for IHC without antigen retrieval? > > Also, thanks to everyone for their suggestions re. mouse B-cell > markers. > > Thanks, > Sharon > -- > Sharon Cooperman > NIH, NICHD, CBMB 301.435-8417 > Building 18T, room 101 301.402-0078 fax > Bethesda, MD 20892 > > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lsb <@t> jax.org Fri Nov 26 09:27:33 2004 From: lsb <@t> jax.org (Lesley S. Bechtold) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Necropsies Message-ID: <6.0.3.0.0.20041126102509.02615e40@aretha.jax.org> Hi Everyone, Do any of you have a good job description for the people who do animal necropsies in your organization? Also, are there any professional societies or accreditations or certifications that they can (or should) obtain? Thank you very much! I hope everyone ate enough and then some yesterday! Happy Holidays! Lesley Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191 From tigersnake <@t> ecybermind.net Fri Nov 26 11:25:41 2004 From: tigersnake <@t> ecybermind.net (Paul Howard Lockwood) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Necropsies In-Reply-To: <6.0.3.0.0.20041126102509.02615e40@aretha.jax.org> References: <6.0.3.0.0.20041126102509.02615e40@aretha.jax.org> Message-ID: Dear Lesley The best certificatio program I can direct you to is The American Association For Laboratory Animal Science. There are three levels o certificatio, but these are broad range certifications, and not for Necrospy alone. I don't think is there is such a specific certification. Here's the web site. Http://www.aalas.org/ As for job descriptions, we had three classifications fo employees working in the necropsy lab. Pathologist: Obvisously a DVM with appropriate verterinarian training and certification through the governoring boards. A prosector bs. preferred, but people could be promoted through time and grade. They were the people who actually did the harvesting of organs, blood, and bone marrow, showing any lesions to the doctor, and reporting all findings verbally to the doctor and the technician. The Technicians could be BS or lower. The placed organs in formalin, collected weights of organs, trimmed all unwanted tissue from organ as directed, prepared all slides and other sameple displays or containers. This is a pretty rough idea, and I hope is helps some. If there is any further need for information, please do not hesitate to ask. Sincerely, Paul Lockwood, HT (ASCP) LaTG (AALAS0 Fr 26 Nov 2004 10:27:33 -0500, Lesley S. Bechtold wrote: > Hi Everyone, > > Do any of you have a good job description for the people who do > animal necropsies in your organization? Also, are there any > professional societies or accreditations or certifications that they can > (or should) obtain? Thank you very much! > > I hope everyone ate enough and then some yesterday! Happy > Holidays! > > Lesley > > > > Lesley S. Bechtold > Supervisor, Biological Imaging > The Jackson Laboratory > 600 Main St. > Bar Harbor, ME 04609 > 207-288-6191 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Using Opera's revolutionary e-mail client: http://www.opera.com/m2/ From scoop <@t> mail.nih.gov Fri Nov 26 11:35:49 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] cell markers for mouse kidney cells? Message-ID: Dear Histonetters, I did immunohistochemistry in mouse kidney and found increased levels of a particular protein in certain cells, but I can't figure out what type of cell the staining is in. Previously, another person in my lab looking at the same protein even showed the sections to a renal pathologist and was told that the cell type couldn't be determined in the sections they had (which were just IHC for the specific protein and counterstain). Is there a way to identify specific cell types in kidney using cell type specific markers and does anyone know what they are or have suggestions on what might be useful to try? Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From Ronnie_Houston <@t> bshsi.com Fri Nov 26 11:56:26 2004 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Transcription productivity data Message-ID: <530361BF03351B4CAE5270A05D3037B5036735F6@bsrexms01.BSHSIR.COM> For those of you who have transcription services under you, can you share some productivity data? I am particularly interested in CoPath users, but would appreciate any and all information available. Thanks Ronnie Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 2877972 (804) 2877906 - fax ronnie_houston@bshsi.com ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone, and permanently delete the original e-mail, attachment(s), and any copies. From pruegg <@t> ihctech.net Fri Nov 26 13:44:34 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] cell markers for mouse kidney cells? In-Reply-To: Message-ID: Sharon, An H&E viewed by a good mouse pathologist should be enough to determine what cell type your protein is expressed in, the other option would be to do double IHC labeling but you would have to know what possible cell type it could be to choose the marker. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Sharon Cooperman Sent: Friday, November 26, 2004 10:36 AM To: histonet@Pathology.swmed.edu Subject: [Histonet] cell markers for mouse kidney cells? Dear Histonetters, I did immunohistochemistry in mouse kidney and found increased levels of a particular protein in certain cells, but I can't figure out what type of cell the staining is in. Previously, another person in my lab looking at the same protein even showed the sections to a renal pathologist and was told that the cell type couldn't be determined in the sections they had (which were just IHC for the specific protein and counterstain). Is there a way to identify specific cell types in kidney using cell type specific markers and does anyone know what they are or have suggestions on what might be useful to try? Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From 9msm8 <@t> qlink.queensu.ca Fri Nov 26 16:12:03 2004 From: 9msm8 <@t> qlink.queensu.ca (9msm8@qlink.queensu.ca) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] H&E water contamination Message-ID: <43526.128.100.240.244.1101507123.squirrel@128.100.240.244> Hi histonet I have been performing H&E stains on sliced parraffin embedded mouse hippocampal slices. I am consistently getting water contamination in my slides and have been unable to rectify the problem. My procedure for staining is: Xylene I for 7.5 minutes; Xylene II for 7.5 minutes; 100% ethanol I for 2 minutes; 100% ethanol II for 2 minutes; 95% ethanol I and 70% ethanol I for 2 minutes each; Water I for a second; then "H" for 20 minutes; Water II for 10 minutes; "E" for about 13 seconds, Water III for about 1 minute; 70% ethanol II for 50 seconds; 95% ethanol II for 5 minutes; 100% ethanol III and IV for 5 minutes each and then Xylene III and IV for 7.5 minutes each. I beleive the "70% ethanol II" is the key step for removing water after the staining but increasing the amount of time spent in the solution hasn't worked yet (though I have yet to leave it in for longer then a minute). Any tips would be greatly appreciated, Mike From contact <@t> excaliburpathology.com Sat Nov 27 12:32:07 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Water contamination Message-ID: <20041127183207.76948.qmail@web50306.mail.yahoo.com> First, you SLICE meat. Histotechs SECTION tissue. 70% EtOH is 30% water. You are probably getting too much water carryover into your 100%. Make sure the 100% EtOH is 100%. If the solutions are fresh, 10 to 20 dips each is enough time for dehydration before xylene and coverslipping. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From AnthonyH <@t> chw.edu.au Sun Nov 28 17:00:02 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] H&E water contamination Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E123@simba.kids> A few ideas. Check your xylene for water. Keep the alcohol and xylene containers covered when not in use. Alcohol will absorb water from the air (possibly xylene as well - though of this I am not certain). 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: 9msm8@qlink.queensu.ca [mailto:9msm8@qlink.queensu.ca] Sent: Saturday, 27 November 2004 9:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] H&E water contamination Hi histonet I have been performing H&E stains on sliced parraffin embedded mouse hippocampal slices. I am consistently getting water contamination in my slides and have been unable to rectify the problem. My procedure for staining is: Xylene I for 7.5 minutes; Xylene II for 7.5 minutes; 100% ethanol I for 2 minutes; 100% ethanol II for 2 minutes; 95% ethanol I and 70% ethanol I for 2 minutes each; Water I for a second; then "H" for 20 minutes; Water II for 10 minutes; "E" for about 13 seconds, Water III for about 1 minute; 70% ethanol II for 50 seconds; 95% ethanol II for 5 minutes; 100% ethanol III and IV for 5 minutes each and then Xylene III and IV for 7.5 minutes each. I beleive the "70% ethanol II" is the key step for removing water after the staining but increasing the amount of time spent in the solution hasn't worked yet (though I have yet to leave it in for longer then a minute). Any tips would be greatly appreciated, Mike _______________________________________________ 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 Kemlo.Rogerson <@t> elht.nhs.uk Mon Nov 29 01:48:46 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] H&E water contamination[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B53C@bhrv-nt-11.bhrv.nwest.nhs.uk> How does the water contamination manifest itself? Do you see water droplets under the scope or do the mounting media just go cloudy? I'm surprised with the length of time you leave them in the alcohols that you have any problems, but personally I'd replace the 70% with pure alcohol, as it would quickly get contaminated with water and I'm not sure that gradations of alcohol of ascending purity make any difference. I assume the slides are not cramped into the racks and that there is agitation. If you use some of the propriety mounting media some are hydroscopic and go cloudy with time, in my experience. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: 9msm8@qlink.queensu.ca [mailto:9msm8@qlink.queensu.ca] Sent: 26 November 2004 22:12 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] H&E water contamination[Scanned] Hi histonet I have been performing H&E stains on sliced parraffin embedded mouse hippocampal slices. I am consistently getting water contamination in my slides and have been unable to rectify the problem. My procedure for staining is: Xylene I for 7.5 minutes; Xylene II for 7.5 minutes; 100% ethanol I for 2 minutes; 100% ethanol II for 2 minutes; 95% ethanol I and 70% ethanol I for 2 minutes each; Water I for a second; then "H" for 20 minutes; Water II for 10 minutes; "E" for about 13 seconds, Water III for about 1 minute; 70% ethanol II for 50 seconds; 95% ethanol II for 5 minutes; 100% ethanol III and IV for 5 minutes each and then Xylene III and IV for 7.5 minutes each. I beleive the "70% ethanol II" is the key step for removing water after the staining but increasing the amount of time spent in the solution hasn't worked yet (though I have yet to leave it in for longer then a minute). Any tips would be greatly appreciated, Mike _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Nov 29 02:02:31 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] H&E water contamination[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B53D@bhrv-nt-11.bhrv.nwest.nhs.uk> Plus xylene will tolerate low levels of water (very low) without going cloudy; the contamination only is seen microscopically as a fine 'mist' of droplets. Knew a Chief Technician that kept anhydrous copper sulphate in his last alcohols to absorb any water carry over. If you try staining in very cold and damp conditions then a mist can form from one alcohol to another. It's something to do with the eutectic point of water or I may just be making that up. All I know is, even with fresh alcohols, staining in cold damp conditions can result in water from the atmosphere contamination the xylenes. The only way I could get round it was to use a hair dryer to raise the temperature a bit, and hope I didn't catch fire. I hasten to add I did that before CPA, etc., and not in my present employ. Those were the good old days of sharpening knives with diamond paste and a bit of wood, and making up your own stains and ripening the haematoxylin in the window......... I must be old. I AM old!!!! 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: 28 November 2004 23:00 To: 9msm8@qlink.queensu.ca; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E water contamination[Scanned] A few ideas. Check your xylene for water. Keep the alcohol and xylene containers covered when not in use. Alcohol will absorb water from the air (possibly xylene as well - though of this I am not certain). 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: 9msm8@qlink.queensu.ca [mailto:9msm8@qlink.queensu.ca] Sent: Saturday, 27 November 2004 9:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] H&E water contamination Hi histonet I have been performing H&E stains on sliced parraffin embedded mouse hippocampal slices. I am consistently getting water contamination in my slides and have been unable to rectify the problem. My procedure for staining is: Xylene I for 7.5 minutes; Xylene II for 7.5 minutes; 100% ethanol I for 2 minutes; 100% ethanol II for 2 minutes; 95% ethanol I and 70% ethanol I for 2 minutes each; Water I for a second; then "H" for 20 minutes; Water II for 10 minutes; "E" for about 13 seconds, Water III for about 1 minute; 70% ethanol II for 50 seconds; 95% ethanol II for 5 minutes; 100% ethanol III and IV for 5 minutes each and then Xylene III and IV for 7.5 minutes each. I beleive the "70% ethanol II" is the key step for removing water after the staining but increasing the amount of time spent in the solution hasn't worked yet (though I have yet to leave it in for longer then a minute). Any tips would be greatly appreciated, Mike _______________________________________________ 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 mcauliff <@t> umdnj.edu Mon Nov 29 12:10:42 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] cell markers for mouse kidney cells? In-Reply-To: References: Message-ID: <41AB6622.3010509@umdnj.edu> Hi Sharon: Kidney histology is not that difficult so I don't know why identifying cells should be a problem. Mouse kidney does not look all that different from human kidney. Are we talking about tubules in the cortex or the medulla, blood vessels, glomeruli or interstitial tissue? Someone else suggested comparing an adjacent H&E section to the IHC section, that seems like the first place to start. If you go to your library and get a copy of "The Kidney" by Brenner and Rector it has a big chapter on kidney anatomy in vol. 1. Also, the "Handbook of Physiology" has a chapter on kidney anatomy. I did my dissertation on kidney and I teach histology so if you will send me a jpeg image I will see if I can figure it out. Geoff Sharon Cooperman wrote: > Dear Histonetters, > > I did immunohistochemistry in mouse kidney and found increased levels > of a particular protein in certain cells, but I can't figure out what > type of cell the staining is in. Previously, another person in my lab > looking at the same protein even showed the sections to a renal > pathologist and was told that the cell type couldn't be determined in > the sections they had (which were just IHC for the specific protein > and counterstain). Is there a way to identify specific cell types in > kidney using cell type specific markers and does anyone know what they > are or have suggestions on what might be useful to try? > > Thanks, > Sharon -- -- ********************************************** 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 jluis.palazon <@t> icman.csic.es Mon Nov 29 10:58:43 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Water contamination Message-ID: <20041129165843.5AF4D4339DE@perceval.uca.es> In my lab we normally use 95 % EtOH before absolute alcohols, instead of the 70 % you use, without any problem. Maybe your contaminating agent is 70 % EtOH, try changing this step with a 95 % one Jos? Luis El dia 27/11/2004 19:32 usted envio el siguiente mensaje: >Date: 27 de Noviembre de 2004 19:32:07 >From: Paula Pierce >Subject: [Histonet] Water contamination >To: histonet@lists.utsouthwestern.edu > > First, you SLICE meat. Histotechs SECTION tissue. > > 70% EtOH is 30% water. You are probably getting too much water carryover into your 100%. Make sure the 100% EtOH is 100%. If the solutions are fresh, 10 to 20 dips each is enough time for dehydration before xylene and coverslipping. > > > 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 > Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From fredrik <@t> u.washington.edu Mon Nov 29 11:52:03 2004 From: fredrik <@t> u.washington.edu (Fredrik Johansson) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Counterstain for Von Kossa Message-ID: <6090EE6D-422F-11D9-B10A-000A95F0ED90@u.washington.edu> Hi, ? I am going to stain 5 um paraffin sections of mouse arteries for calcium (Von Kossa) and need a counterstain. I?m new in this area and would appreciate if some one could tell me were to find a protocol for this. I have looked in the archive and found a protocol for bone sections (toluidine blue) can I use the same protocol for my mouse arteries? ? Thanks ? Fredrik Johansson Department of Pathology UW Seattle From liz <@t> premierlab.com Mon Nov 29 12:24:27 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Counterstain for Von Kossa In-Reply-To: <6090EE6D-422F-11D9-B10A-000A95F0ED90@u.washington.edu> Message-ID: <000401c4d640$a973dc10$76d48a80@AMY> Frederik Nuclear Fast Red is the counterstain that I use when I run a Von Kossa. 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 Fredrik Johansson Sent: Monday, November 29, 2004 10:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Counterstain for Von Kossa Hi, ? I am going to stain 5 um paraffin sections of mouse arteries for calcium (Von Kossa) and need a counterstain. I?m new in this area and would appreciate if some one could tell me were to find a protocol for this. I have looked in the archive and found a protocol for bone sections (toluidine blue) can I use the same protocol for my mouse arteries? ? Thanks ? Fredrik Johansson Department of Pathology UW Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From STapper <@t> slhduluth.com Mon Nov 29 12:33:12 2004 From: STapper <@t> slhduluth.com (Tapper, Sheila) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] automated glass coverslippers Message-ID: <3BFBBD68413CB443A7125A63EC0ACD1D33A62D@slhw2smail01.slhdomain.com> I am wondering if anyone has the Sakura Glass coverslipper, and if they are experiencing any significant troubles? What is everyone else using (for glass) out there? You may want to answer directly to me so that we don't clog the 'net with my silly question!! Thanks! Sheila CONFIDENTIAL: The confidential information accompanying this transmission may contain protected health information and is legally privileged under state and federal law. This information is intended only for the use of the individual to which it is addressed. The recipient or person responsible for delivering this information is prohibited by law from disclosing this information without proper authorization to any other party, unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any review, 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. No response indicates that the information was received by the appropriate authorized party. Warning: Although St. Luke's has taken reasonable precautions to ensure no viruses are present in this email, St. Luke's cannot accept responsibility for any loss or damage arising from the use of this email or attachments. From contact <@t> excaliburpathology.com Mon Nov 29 12:34:59 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Counterstain for Von Kossa Message-ID: <20041129183459.61709.qmail@web50310.mail.yahoo.com> Any Histology staining book will have a procedure for this stain. Lots of counterstains to choose from, but nuclear fast red is the standard. I am going to rant here, so please be patient with me. How many places out there are opening up labs or doing research with no trained people? And if these new people have internet access to find the histonet, they should be finding websites with actual procedures, AND most colleges and universities have libraries with REAL books on histology procedures. Go there and read. Since becoming registered 25 years ago, when I was asked to do something new I would go to my own library I have accumulated or the university library if it was really unusual and read, because we did not have the internet. From JMacDonald <@t> mtsac.edu Mon Nov 29 12:52:02 2004 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] H&E water contamination[Scanned] In-Reply-To: <1030B679AD69D6119C3F00080210DD9D01B0B53D@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: We recently discovered that all of our "absolute" alcohol contains water. You could have a contaminated lot. Jennifer MacDonald Kemlo Rogerson Sent by: histonet-bounces@lists.utsouthwestern.edu 11/29/2004 12:02 AM To histonet@lists.utsouthwestern.edu cc Subject RE: [Histonet] H&E water contamination[Scanned] Plus xylene will tolerate low levels of water (very low) without going cloudy; the contamination only is seen microscopically as a fine 'mist' of droplets. Knew a Chief Technician that kept anhydrous copper sulphate in his last alcohols to absorb any water carry over. If you try staining in very cold and damp conditions then a mist can form from one alcohol to another. It's something to do with the eutectic point of water or I may just be making that up. All I know is, even with fresh alcohols, staining in cold damp conditions can result in water from the atmosphere contamination the xylenes. The only way I could get round it was to use a hair dryer to raise the temperature a bit, and hope I didn't catch fire. I hasten to add I did that before CPA, etc., and not in my present employ. Those were the good old days of sharpening knives with diamond paste and a bit of wood, and making up your own stains and ripening the haematoxylin in the window......... I must be old. I AM old!!!! 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: 28 November 2004 23:00 To: 9msm8@qlink.queensu.ca; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] H&E water contamination[Scanned] A few ideas. Check your xylene for water. Keep the alcohol and xylene containers covered when not in use. Alcohol will absorb water from the air (possibly xylene as well - though of this I am not certain). 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: 9msm8@qlink.queensu.ca [mailto:9msm8@qlink.queensu.ca] Sent: Saturday, 27 November 2004 9:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] H&E water contamination Hi histonet I have been performing H&E stains on sliced parraffin embedded mouse hippocampal slices. I am consistently getting water contamination in my slides and have been unable to rectify the problem. My procedure for staining is: Xylene I for 7.5 minutes; Xylene II for 7.5 minutes; 100% ethanol I for 2 minutes; 100% ethanol II for 2 minutes; 95% ethanol I and 70% ethanol I for 2 minutes each; Water I for a second; then "H" for 20 minutes; Water II for 10 minutes; "E" for about 13 seconds, Water III for about 1 minute; 70% ethanol II for 50 seconds; 95% ethanol II for 5 minutes; 100% ethanol III and IV for 5 minutes each and then Xylene III and IV for 7.5 minutes each. I beleive the "70% ethanol II" is the key step for removing water after the staining but increasing the amount of time spent in the solution hasn't worked yet (though I have yet to leave it in for longer then a minute). Any tips would be greatly appreciated, Mike _______________________________________________ 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Linke_Noelle <@t> Allergan.com Mon Nov 29 13:00:44 2004 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] automated glass coverslippers Message-ID: This is not a silly question. The Sakura Glas coverslipper, in my honest and humble opinion is a giant hunk of junk. It gets goop all over the back of the slides unless you clean the dispensing tip with Xylene every 5 seconds, the belt jams, and the very expensive German coverslips that Sakura recommends do not help with any of those problems! I would not cry a tear if it broke and needed to be replaced! Noelle Linke, BS, HTL(ASCP)QIHC Allergan, Inc 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5568 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tapper, Sheila Sent: Monday, November 29, 2004 10:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] automated glass coverslippers I am wondering if anyone has the Sakura Glass coverslipper, and if they are experiencing any significant troubles? What is everyone else using (for glass) out there? You may want to answer directly to me so that we don't clog the 'net with my silly question!! Thanks! Sheila CONFIDENTIAL: The confidential information accompanying this transmission may contain protected health information and is legally privileged under state and federal law. This information is intended only for the use of the individual to which it is addressed. The recipient or person responsible for delivering this information is prohibited by law from disclosing this information without proper authorization to any other party, unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any review, 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. No response indicates that the information was received by the appropriate authorized party. Warning: Although St. Luke's has taken reasonable precautions to ensure no viruses are present in this email, St. Luke's cannot accept responsibility for any loss or damage arising from the use of this email or attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From convmcm <@t> cc.usu.edu Mon Nov 29 13:24:47 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Counterstain for Von Kossa In-Reply-To: <20041129183459.61709.qmail@web50310.mail.yahoo.com> Message-ID: <000401c4d649$169fe1f0$4a737b81@Cygnus> I think there is a really valuable learning experience in going to libraries and doing a web search before asking question on this list. I have always found that digging out info on my own helps me learn thing quicker and faster than having it spoon fed. Having said that, I also believe that this list provides a valuable reference for those whose time may not allow them the luxury of going to the library or making an extensive internet search. I, for one, am happy to provide info to anyone who asks. My view is... if anyone feels the requested info is too basic to bother with, don't bother replying. Connie McManus email: convmcm@cc.usu.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Pierce Sent: Monday, November 29, 2004 11:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Counterstain for Von Kossa Any Histology staining book will have a procedure for this stain. Lots of counterstains to choose from, but nuclear fast red is the standard. I am going to rant here, so please be patient with me. How many places out there are opening up labs or doing research with no trained people? And if these new people have internet access to find the histonet, they should be finding websites with actual procedures, AND most colleges and universities have libraries with REAL books on histology procedures. Go there and read. Since becoming registered 25 years ago, when I was asked to do something new I would go to my own library I have accumulated or the university library if it was really unusual and read, because we did not have the internet. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Mon Nov 29 13:38:24 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] serum amyloid a - saa1 immunohistochemistry in mouse Message-ID: <000001c4d64a$fdc86830$76d48a80@AMY> Hello Everyone I have a question regarding detection of serum amyloid a (saa1) via immunohistochemistry in mouse tissues. I have searched the histonet archives and have seen one response that Dako's Amyloid A antibody will work in mouse tissues. This is a mouse monoclonal antibody. When I searched pub med there were several references to a rabbit anti-mouse SAA1 antibody. I have been not been able to locate a vendor for this rabbit anti-mouse SAA1. Is anybody aware of a vendor that sells that antibody or any other antibody that will react with mouse saa1. Thanks in advance. 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 vazquezr <@t> ohsu.edu Mon Nov 29 13:54:43 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Counterstain for Von Kossa Message-ID: I agree with Connie, there is no stupid question. People will stop asking questions, if someone/themselves gets chastised every time someone/themselves asks a insignificant question. This forum is not just all about antibodies. Robyn OHSU >>> "Connie McManus" 11/29/2004 11:24:47 AM >>> I think there is a really valuable learning experience in going to libraries and doing a web search before asking question on this list. I have always found that digging out info on my own helps me learn thing quicker and faster than having it spoon fed. Having said that, I also believe that this list provides a valuable reference for those whose time may not allow them the luxury of going to the library or making an extensive internet search. I, for one, am happy to provide info to anyone who asks. My view is... if anyone feels the requested info is too basic to bother with, don't bother replying. Connie McManus email: convmcm@cc.usu.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Pierce Sent: Monday, November 29, 2004 11:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Counterstain for Von Kossa Any Histology staining book will have a procedure for this stain. Lots of counterstains to choose from, but nuclear fast red is the standard. I am going to rant here, so please be patient with me. How many places out there are opening up labs or doing research with no trained people? And if these new people have internet access to find the histonet, they should be finding websites with actual procedures, AND most colleges and universities have libraries with REAL books on histology procedures. Go there and read. Since becoming registered 25 years ago, when I was asked to do something new I would go to my own library I have accumulated or the university library if it was really unusual and read, because we did not have the internet. _______________________________________________ Histonet mailing list Histonet@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 darrenj <@t> medica.co.nz Mon Nov 29 14:57:57 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Counterstain for Von Kossa In-Reply-To: <000401c4d649$169fe1f0$4a737b81@Cygnus> Message-ID: <000101c4d656$1b737720$c864a8c0@medica.co.nz> I agree with Connie. I personally would research something myself prior to asking in this forum and would always suggest to co-workers (particularly trainees) that they do the same. If you take the time to look something up and research it I think you do learn more. However I think that recent comments implying that people who ask "easy" questions are either lazy or not appropriately qualified are perhaps a little unnecessary. One person's "easy" question may cause a melt down for another. Surely the whole aim of his forum is sharing knowledge and experience, not judging others. Some people may not have a reference library or have the luxury of being able to sit on the net for hours trying to find out information related to a stain or procedure. Also those who work in a high pressure environment may not have the time or resources to test and trial new methods but would prefer to use an already "perfected" method from another institution. Perhaps we should show a little professional courtesy to our colleagues and give them the benefit of the doubt. I will get down from my soapbox now before I fall off (or am pushed). In regards to the original counterstain question, I always used Tol Blue when I performed Von Kossa on undecalcified bone sections. Have a great day Darren James -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Connie McManus Sent: Tuesday, 30 November 2004 8:25 a.m. To: 'Paula Pierce'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Counterstain for Von Kossa I think there is a really valuable learning experience in going to libraries and doing a web search before asking question on this list. I have always found that digging out info on my own helps me learn thing quicker and faster than having it spoon fed. Having said that, I also believe that this list provides a valuable reference for those whose time may not allow them the luxury of going to the library or making an extensive internet search. I, for one, am happy to provide info to anyone who asks. My view is... if anyone feels the requested info is too basic to bother with, don't bother replying. Connie McManus email: convmcm@cc.usu.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Pierce Sent: Monday, November 29, 2004 11:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Counterstain for Von Kossa Any Histology staining book will have a procedure for this stain. Lots of counterstains to choose from, but nuclear fast red is the standard. I am going to rant here, so please be patient with me. How many places out there are opening up labs or doing research with no trained people? And if these new people have internet access to find the histonet, they should be finding websites with actual procedures, AND most colleges and universities have libraries with REAL books on histology procedures. Go there and read. Since becoming registered 25 years ago, when I was asked to do something new I would go to my own library I have accumulated or the university library if it was really unusual and read, because we did not have the internet. _______________________________________________ Histonet mailing list Histonet@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 hymclab <@t> hyhc.com Mon Nov 29 15:30:50 2004 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] RE: IHC on tissues processed for EM Message-ID: I too think that no question should be lableled as "easy". There are no stupid questions. I recently went through an issue about a stain that I have been doing for over 20 years. Couldn't figure out what was wrong. Technical services at two companies were stumped until someone came up with a possible answer. I tested it and they were correct. Someone else on the Histonetlist just had the same problem and I responded with what we found to be our problem. Then I got slammed by someone who couldn't have even read the whole message. I was answering someone'e problem about their mucicarmine stain not working. In my response to this person I used the terms Mayer's mucicarmine and Weigert's Hematoxylin. This other person that slammed accused me in living in the middle ages for using Mayer's Hematoxlyin and Weigert's hematoxlyin and said I should try Gill's hematoxylin. I didn't say I used Mayer's hematoxylin, I was referring to the name of the Mucicarmine stain. Secondly I had never heard of using Gill's in place of Weigert's in the Mucicarmine stain. I e-mailed him back I thought maybe he was confused---he said he wasn't and he said that he'd never even done a Mucicarmine stain. I rest my case. Things like that make you want to be a lurker instead of trying to help someone. Dawn -----Original Message----- From: Gudrun Lang [mailto:gu.lang@gmx.at] Sent: Thursday, November 25, 2004 3:09 AM To: Histonetliste Subject: Re: [Histonet] RE: IHC on tissues processed for EM Laszlo, I feel about saying some words about your message. You complain, that members of the histonetlist have no basic knowledgment. But what is basic? For me, in a routine surgery histolab, EM is very, very special for example. I belong to those people, that work in a small corner of the whole histological techniques. But I am interested and want to learn. You say, people have to use databases, text books etc. In my case I have no access through my lab to those things and they are'nt cheap (and first you have to know where to look). I am very happy about the histonetlist. Here are specialists, experts from every field and also "workmates". Please be tolerant with "easy" questions and problems, perhaps the person considers the problem more difficult than you know. friendly greetings Gudrun Lang ----- Original Message ----- From: "Laszlo Komuves" To: Sent: Wednesday, November 24, 2004 8:44 PM Subject: [Histonet] RE: IHC on tissues processed for EM There are hundreds, if not thousands of papers demonstrating successful localization of intracellular antigens at EM or LM level using tissue samples processed conventionally for EM (i.e. PFA/GA fixation, Os postfixation, epoxy resin embedding). Look for the now classical papers by Roth and Bendayan or their early reviews. Also the EM textbooks published in the 80's have plenty of information. And let me also add a personal comment of frustration: Histonet used to be about sharing experience and wisdom, mentoring and teaching those have were (are they still?) eager to learn or perfect their skills. I personally learnt/benefited from many messages posted. Now people are asking advice about coverslip sizes? So thanks to the curse of the Internet, journal articles and text books are used only by a dedicated few, electronic databases (libraries, PubMed, journal archives, reagent search sites, even Google) are not searched, and now even the vendor catalogs are not opened, because on the Histonet somebody will respond? How will users differentiate between solid information (based on scientific knowledge, practical experience) or just plain ignorance? Where is critical thinking? What happened to a once honorable guild of skilled craftsmanship, when members of the community are lacking elementary knowledge (just a few recent hair-raising topics: H&E staining, pH, and the list goes on and on)? And of course we/they are constantly offended/complain about the lack of recognition by our/their peers. Feel free to get angry with me. I certainly welcome arguments and criticism.. As for me, I go back to find something useful in one of my textbooks. Laszlo Komuves FROM: L?szl? G. Komuves PhD Senior Principal Scientist, Manager, Microscopy Core Laboratory CORGENTECH, Inc. 650 Gateway Blvd., South San Francisco, CA 94080 Phone: (650) 246-6905, Fax: (650) 624-7540 E-mail: komuves@corgentech.com Date: Tue, 23 Nov 2004 08:02:24 -0500 From: Michele French Subject: [Histonet] IHC on Sections Processed for EM? To: histonet@lists.utsouthwestern.edu Message-ID: <41A334E0.2060308@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Good Morning Histonet! A colleague of mine did some EM work and found something interesting. Our pathologist wanted me to try to do some IHC to further characterize what is present in the section. Our EM person said it would never work. I did not think it was possible either, but I thought I would ask anyway. Has anyone tried doing an immunostain on plastic (Epon) sections from tissue fixed and processed for EM? I am always up for a challenge, but I am really busy right now and don't want to waste my time if there is really no hope! Thanks in advance, Michele French _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Ronnie_Houston <@t> bshsi.com Mon Nov 29 15:37:44 2004 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] RE: IHC on tissues processed for EM Message-ID: <530361BF03351B4CAE5270A05D3037B503673600@bsrexms01.BSHSIR.COM> Now I remember why I sign off the list for extended periods Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 2877972 (804) 2877906 - fax ronnie_houston@bshsi.com -----Original Message----- From: hymclab [mailto:hymclab@hyhc.com] Sent: Monday, November 29, 2004 4:31 PM To: 'Gudrun Lang'; Histonetliste Subject: RE: [Histonet] RE: IHC on tissues processed for EM I too think that no question should be lableled as "easy". There are no stupid questions. I recently went through an issue about a stain that I have been doing for over 20 years. Couldn't figure out what was wrong. Technical services at two companies were stumped until someone came up with a possible answer. I tested it and they were correct. Someone else on the Histonetlist just had the same problem and I responded with what we found to be our problem. Then I got slammed by someone who couldn't have even read the whole message. I was answering someone'e problem about their mucicarmine stain not working. In my response to this person I used the terms Mayer's mucicarmine and Weigert's Hematoxylin. This other person that slammed accused me in living in the middle ages for using Mayer's Hematoxlyin and Weigert's hematoxlyin and said I should try Gill's hematoxylin. I didn't say I used Mayer's hematoxylin, I was referring to the name of the Mucicarmine stain. Secondly I had never heard of using Gill's in place of Weigert's in the Mucicarmine stain. I e-mailed him back I thought maybe he was confused---he said he wasn't and he said that he'd never even done a Mucicarmine stain. I rest my case. Things like that make you want to be a lurker instead of trying to help someone. Dawn -----Original Message----- From: Gudrun Lang [mailto:gu.lang@gmx.at] Sent: Thursday, November 25, 2004 3:09 AM To: Histonetliste Subject: Re: [Histonet] RE: IHC on tissues processed for EM Laszlo, I feel about saying some words about your message. You complain, that members of the histonetlist have no basic knowledgment. But what is basic? For me, in a routine surgery histolab, EM is very, very special for example. I belong to those people, that work in a small corner of the whole histological techniques. But I am interested and want to learn. You say, people have to use databases, text books etc. In my case I have no access through my lab to those things and they are'nt cheap (and first you have to know where to look). I am very happy about the histonetlist. Here are specialists, experts from every field and also "workmates". Please be tolerant with "easy" questions and problems, perhaps the person considers the problem more difficult than you know. friendly greetings Gudrun Lang ----- Original Message ----- From: "Laszlo Komuves" To: Sent: Wednesday, November 24, 2004 8:44 PM Subject: [Histonet] RE: IHC on tissues processed for EM There are hundreds, if not thousands of papers demonstrating successful localization of intracellular antigens at EM or LM level using tissue samples processed conventionally for EM (i.e. PFA/GA fixation, Os postfixation, epoxy resin embedding). Look for the now classical papers by Roth and Bendayan or their early reviews. Also the EM textbooks published in the 80's have plenty of information. And let me also add a personal comment of frustration: Histonet used to be about sharing experience and wisdom, mentoring and teaching those have were (are they still?) eager to learn or perfect their skills. I personally learnt/benefited from many messages posted. Now people are asking advice about coverslip sizes? So thanks to the curse of the Internet, journal articles and text books are used only by a dedicated few, electronic databases (libraries, PubMed, journal archives, reagent search sites, even Google) are not searched, and now even the vendor catalogs are not opened, because on the Histonet somebody will respond? How will users differentiate between solid information (based on scientific knowledge, practical experience) or just plain ignorance? Where is critical thinking? What happened to a once honorable guild of skilled craftsmanship, when members of the community are lacking elementary knowledge (just a few recent hair-raising topics: H&E staining, pH, and the list goes on and on)? And of course we/they are constantly offended/complain about the lack of recognition by our/their peers. Feel free to get angry with me. I certainly welcome arguments and criticism.. As for me, I go back to find something useful in one of my textbooks. Laszlo Komuves FROM: L?szl? G. Komuves PhD Senior Principal Scientist, Manager, Microscopy Core Laboratory CORGENTECH, Inc. 650 Gateway Blvd., South San Francisco, CA 94080 Phone: (650) 246-6905, Fax: (650) 624-7540 E-mail: komuves@corgentech.com Date: Tue, 23 Nov 2004 08:02:24 -0500 From: Michele French Subject: [Histonet] IHC on Sections Processed for EM? To: histonet@lists.utsouthwestern.edu Message-ID: <41A334E0.2060308@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Good Morning Histonet! A colleague of mine did some EM work and found something interesting. Our pathologist wanted me to try to do some IHC to further characterize what is present in the section. Our EM person said it would never work. I did not think it was possible either, but I thought I would ask anyway. Has anyone tried doing an immunostain on plastic (Epon) sections from tissue fixed and processed for EM? I am always up for a challenge, but I am really busy right now and don't want to waste my time if there is really no hope! Thanks in advance, Michele French _______________________________________________ Histonet mailing list Histonet@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 ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone, and permanently delete the original e-mail, attachment(s), and any copies. From AnthonyH <@t> chw.edu.au Mon Nov 29 15:52:51 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Counterstain for Von Kossa Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E125@simba.kids> For Von Kossa, there are many counterstains you can use: Nuclear Fast Red, Ethyl (was Methyl) Green (pH 4.2), even a light eosin. Take your pick. What other structures do you want to co-observe (Is that a word?)? 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: Fredrik Johansson [mailto:fredrik@u.washington.edu] Sent: Tuesday, 30 November 2004 4:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Counterstain for Von Kossa Hi, ? I am going to stain 5 um paraffin sections of mouse arteries for calcium (Von Kossa) and need a counterstain. I'm new in this area and would appreciate if some one could tell me were to find a protocol for this. I have looked in the archive and found a protocol for bone sections (toluidine blue) can I use the same protocol for my mouse arteries? ? Thanks ? Fredrik Johansson Department of Pathology UW Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From AnthonyH <@t> chw.edu.au Mon Nov 29 16:01:41 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] RE: IHC on tissues processed for EM Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E126@simba.kids> I agree with you. If you don't use Gill's - why are you in the Middle Ages? I personally have used Gill's (a commercial formulation) and was not impressed. In bowel sections, I had the mucin staining and after only a few days both haematoxylin and alum precipitate. Performance was definitly not up to scratch compared to the Modified Harris's we were using at the time. This was some 10 years ago, so maybe the commercial formulations have improved (I hope). My recommendation is to use the techniques and stains that suit your laboratory and meet your QC requirements. Remember we must always be on the look-out for something better. Amen!! 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: hymclab [mailto:hymclab@hyhc.com] Sent: Tuesday, 30 November 2004 8:31 AM To: 'Gudrun Lang'; Histonetliste Subject: RE: [Histonet] RE: IHC on tissues processed for EM I too think that no question should be lableled as "easy". There are no stupid questions. I recently went through an issue about a stain that I have been doing for over 20 years. Couldn't figure out what was wrong. Technical services at two companies were stumped until someone came up with a possible answer. I tested it and they were correct. Someone else on the Histonetlist just had the same problem and I responded with what we found to be our problem. Then I got slammed by someone who couldn't have even read the whole message. I was answering someone'e problem about their mucicarmine stain not working. In my response to this person I used the terms Mayer's mucicarmine and Weigert's Hematoxylin. This other person that slammed accused me in living in the middle ages for using Mayer's Hematoxlyin and Weigert's hematoxlyin and said I should try Gill's hematoxylin. I didn't say I used Mayer's hematoxylin, I was referring to the name of the Mucicarmine stain. Secondly I had never heard of using Gill's in place of Weigert's in the Mucicarmine stain. I e-mailed him back I thought maybe he was confused---he said he wasn't and he said that he'd never even done a Mucicarmine stain. I rest my case. Things like that make you want to be a lurker instead of trying to help someone. Dawn -----Original Message----- From: Gudrun Lang [mailto:gu.lang@gmx.at] Sent: Thursday, November 25, 2004 3:09 AM To: Histonetliste Subject: Re: [Histonet] RE: IHC on tissues processed for EM Laszlo, I feel about saying some words about your message. You complain, that members of the histonetlist have no basic knowledgment. But what is basic? For me, in a routine surgery histolab, EM is very, very special for example. I belong to those people, that work in a small corner of the whole histological techniques. But I am interested and want to learn. You say, people have to use databases, text books etc. In my case I have no access through my lab to those things and they are'nt cheap (and first you have to know where to look). I am very happy about the histonetlist. Here are specialists, experts from every field and also "workmates". Please be tolerant with "easy" questions and problems, perhaps the person considers the problem more difficult than you know. friendly greetings Gudrun Lang ----- Original Message ----- From: "Laszlo Komuves" To: Sent: Wednesday, November 24, 2004 8:44 PM Subject: [Histonet] RE: IHC on tissues processed for EM There are hundreds, if not thousands of papers demonstrating successful localization of intracellular antigens at EM or LM level using tissue samples processed conventionally for EM (i.e. PFA/GA fixation, Os postfixation, epoxy resin embedding). Look for the now classical papers by Roth and Bendayan or their early reviews. Also the EM textbooks published in the 80's have plenty of information. And let me also add a personal comment of frustration: Histonet used to be about sharing experience and wisdom, mentoring and teaching those have were (are they still?) eager to learn or perfect their skills. I personally learnt/benefited from many messages posted. Now people are asking advice about coverslip sizes? So thanks to the curse of the Internet, journal articles and text books are used only by a dedicated few, electronic databases (libraries, PubMed, journal archives, reagent search sites, even Google) are not searched, and now even the vendor catalogs are not opened, because on the Histonet somebody will respond? How will users differentiate between solid information (based on scientific knowledge, practical experience) or just plain ignorance? Where is critical thinking? What happened to a once honorable guild of skilled craftsmanship, when members of the community are lacking elementary knowledge (just a few recent hair-raising topics: H&E staining, pH, and the list goes on and on)? And of course we/they are constantly offended/complain about the lack of recognition by our/their peers. Feel free to get angry with me. I certainly welcome arguments and criticism.. As for me, I go back to find something useful in one of my textbooks. Laszlo Komuves FROM: L?szl? G. Komuves PhD Senior Principal Scientist, Manager, Microscopy Core Laboratory CORGENTECH, Inc. 650 Gateway Blvd., South San Francisco, CA 94080 Phone: (650) 246-6905, Fax: (650) 624-7540 E-mail: komuves@corgentech.com Date: Tue, 23 Nov 2004 08:02:24 -0500 From: Michele French Subject: [Histonet] IHC on Sections Processed for EM? To: histonet@lists.utsouthwestern.edu Message-ID: <41A334E0.2060308@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Good Morning Histonet! A colleague of mine did some EM work and found something interesting. Our pathologist wanted me to try to do some IHC to further characterize what is present in the section. Our EM person said it would never work. I did not think it was possible either, but I thought I would ask anyway. Has anyone tried doing an immunostain on plastic (Epon) sections from tissue fixed and processed for EM? I am always up for a challenge, but I am really busy right now and don't want to waste my time if there is really no hope! Thanks in advance, Michele French _______________________________________________ Histonet mailing list Histonet@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 ********************************************************************** 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 Barry.R.Rittman <@t> uth.tmc.edu Mon Nov 29 16:10:03 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] RE: IHC on tissues processed for EM Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0010DF141@UTHEVS3.mail.uthouston.edu> Dawn et al. I do not think that this is a totally black and white problem. I think that it is entirely appropriate to ask people on Histonet for their opinion on a specific technique as there are many experts in the various fields. While I agree that there are no stupid questions, I believe that the intent of the writer was not to deride the individual but to suggest that they first try to search out the information before asking the question. I have taught students for many years and feel that more and more I am being asked to spoon feed individuals rather than to get them to learn the material. It is easier to provide the correct answer and considerably more difficult to motivate people to work out the solution for themselves (if they can) i.e. I believe that a ready made answer is usually not in the best interests of learning for that individual. When I was training (and I feel that I still am) while I could go and ask an "expert" next door for the answer, I always tried to find the answer out by myself first. This was, I believe, to my benefit as I was able not only to ask a more pointed question of the expert but also ask some important follow up questions. I believe that it is a much more satisfying approach. On the other side of the coin you have to realize that some of the individuals asking these questions may not have the resources available to them or even know where to begin to find out the information. I would suggest to those who wish to ask questions that you first try to research the material if you can as you will then be able to ask questions specific to your needs. You will benefit both ways. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of hymclab Sent: Monday, November 29, 2004 3:31 PM To: 'Gudrun Lang'; Histonetliste Subject: RE: [Histonet] RE: IHC on tissues processed for EM I too think that no question should be lableled as "easy". There are no stupid questions. I recently went through an issue about a stain that I have been doing for over 20 years. Couldn't figure out what was wrong. Technical services at two companies were stumped until someone came up with a possible answer. I tested it and they were correct. Someone else on the Histonetlist just had the same problem and I responded with what we found to be our problem. Then I got slammed by someone who couldn't have even read the whole message. I was answering someone'e problem about their mucicarmine stain not working. In my response to this person I used the terms Mayer's mucicarmine and Weigert's Hematoxylin. This other person that slammed accused me in living in the middle ages for using Mayer's Hematoxlyin and Weigert's hematoxlyin and said I should try Gill's hematoxylin. I didn't say I used Mayer's hematoxylin, I was referring to the name of the Mucicarmine stain. Secondly I had never heard of using Gill's in place of Weigert's in the Mucicarmine stain. I e-mailed him back I thought maybe he was confused---he said he wasn't and he said that he'd never even done a Mucicarmine stain. I rest my case. Things like that make you want to be a lurker instead of trying to help someone. Dawn -----Original Message----- From: Gudrun Lang [mailto:gu.lang@gmx.at] Sent: Thursday, November 25, 2004 3:09 AM To: Histonetliste Subject: Re: [Histonet] RE: IHC on tissues processed for EM Laszlo, I feel about saying some words about your message. You complain, that members of the histonetlist have no basic knowledgment. But what is basic? For me, in a routine surgery histolab, EM is very, very special for example. I belong to those people, that work in a small corner of the whole histological techniques. But I am interested and want to learn. You say, people have to use databases, text books etc. In my case I have no access through my lab to those things and they are'nt cheap (and first you have to know where to look). I am very happy about the histonetlist. Here are specialists, experts from every field and also "workmates". Please be tolerant with "easy" questions and problems, perhaps the person considers the problem more difficult than you know. friendly greetings Gudrun Lang ----- Original Message ----- From: "Laszlo Komuves" To: Sent: Wednesday, November 24, 2004 8:44 PM Subject: [Histonet] RE: IHC on tissues processed for EM There are hundreds, if not thousands of papers demonstrating successful localization of intracellular antigens at EM or LM level using tissue samples processed conventionally for EM (i.e. PFA/GA fixation, Os postfixation, epoxy resin embedding). Look for the now classical papers by Roth and Bendayan or their early reviews. Also the EM textbooks published in the 80's have plenty of information. And let me also add a personal comment of frustration: Histonet used to be about sharing experience and wisdom, mentoring and teaching those have were (are they still?) eager to learn or perfect their skills. I personally learnt/benefited from many messages posted. Now people are asking advice about coverslip sizes? So thanks to the curse of the Internet, journal articles and text books are used only by a dedicated few, electronic databases (libraries, PubMed, journal archives, reagent search sites, even Google) are not searched, and now even the vendor catalogs are not opened, because on the Histonet somebody will respond? How will users differentiate between solid information (based on scientific knowledge, practical experience) or just plain ignorance? Where is critical thinking? What happened to a once honorable guild of skilled craftsmanship, when members of the community are lacking elementary knowledge (just a few recent hair-raising topics: H&E staining, pH, and the list goes on and on)? And of course we/they are constantly offended/complain about the lack of recognition by our/their peers. Feel free to get angry with me. I certainly welcome arguments and criticism.. As for me, I go back to find something useful in one of my textbooks. Laszlo Komuves FROM: L?szl? G. Komuves PhD Senior Principal Scientist, Manager, Microscopy Core Laboratory CORGENTECH, Inc. 650 Gateway Blvd., South San Francisco, CA 94080 Phone: (650) 246-6905, Fax: (650) 624-7540 E-mail: komuves@corgentech.com Date: Tue, 23 Nov 2004 08:02:24 -0500 From: Michele French Subject: [Histonet] IHC on Sections Processed for EM? To: histonet@lists.utsouthwestern.edu Message-ID: <41A334E0.2060308@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Good Morning Histonet! A colleague of mine did some EM work and found something interesting. Our pathologist wanted me to try to do some IHC to further characterize what is present in the section. Our EM person said it would never work. I did not think it was possible either, but I thought I would ask anyway. Has anyone tried doing an immunostain on plastic (Epon) sections from tissue fixed and processed for EM? I am always up for a challenge, but I am really busy right now and don't want to waste my time if there is really no hope! Thanks in advance, Michele French _______________________________________________ Histonet mailing list Histonet@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 Mon Nov 29 16:18:18 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] RE: IHC on tissues processed for EM Message-ID: Bravo for speaking up!!! Robyn >>> hymclab 11/29/2004 1:30:50 PM >>> I too think that no question should be lableled as "easy". There are no stupid questions. I recently went through an issue about a stain that I have been doing for over 20 years. Couldn't figure out what was wrong. Technical services at two companies were stumped until someone came up with a possible answer. I tested it and they were correct. Someone else on the Histonetlist just had the same problem and I responded with what we found to be our problem. Then I got slammed by someone who couldn't have even read the whole message. I was answering someone'e problem about their mucicarmine stain not working. In my response to this person I used the terms Mayer's mucicarmine and Weigert's Hematoxylin. This other person that slammed accused me in living in the middle ages for using Mayer's Hematoxlyin and Weigert's hematoxlyin and said I should try Gill's hematoxylin. I didn't say I used Mayer's hematoxylin, I was referring to the name of the Mucicarmine stain. Secondly I had never heard of using Gill's in place of Weigert's in the Mucicarmine stain. I e-mailed him back I thought maybe he was confused---he said he wasn't and he said that he'd never even done a Mucicarmine stain. I rest my case. Things like that make you want to be a lurker instead of trying to help someone. Dawn -----Original Message----- From: Gudrun Lang [mailto:gu.lang@gmx.at] Sent: Thursday, November 25, 2004 3:09 AM To: Histonetliste Subject: Re: [Histonet] RE: IHC on tissues processed for EM Laszlo, I feel about saying some words about your message. You complain, that members of the histonetlist have no basic knowledgment. But what is basic? For me, in a routine surgery histolab, EM is very, very special for example. I belong to those people, that work in a small corner of the whole histological techniques. But I am interested and want to learn. You say, people have to use databases, text books etc. In my case I have no access through my lab to those things and they are'nt cheap (and first you have to know where to look). I am very happy about the histonetlist. Here are specialists, experts from every field and also "workmates". Please be tolerant with "easy" questions and problems, perhaps the person considers the problem more difficult than you know. friendly greetings Gudrun Lang ----- Original Message ----- From: "Laszlo Komuves" To: Sent: Wednesday, November 24, 2004 8:44 PM Subject: [Histonet] RE: IHC on tissues processed for EM There are hundreds, if not thousands of papers demonstrating successful localization of intracellular antigens at EM or LM level using tissue samples processed conventionally for EM (i.e. PFA/GA fixation, Os postfixation, epoxy resin embedding). Look for the now classical papers by Roth and Bendayan or their early reviews. Also the EM textbooks published in the 80's have plenty of information. And let me also add a personal comment of frustration: Histonet used to be about sharing experience and wisdom, mentoring and teaching those have were (are they still?) eager to learn or perfect their skills. I personally learnt/benefited from many messages posted. Now people are asking advice about coverslip sizes? So thanks to the curse of the Internet, journal articles and text books are used only by a dedicated few, electronic databases (libraries, PubMed, journal archives, reagent search sites, even Google) are not searched, and now even the vendor catalogs are not opened, because on the Histonet somebody will respond? How will users differentiate between solid information (based on scientific knowledge, practical experience) or just plain ignorance? Where is critical thinking? What happened to a once honorable guild of skilled craftsmanship, when members of the community are lacking elementary knowledge (just a few recent hair-raising topics: H&E staining, pH, and the list goes on and on)? And of course we/they are constantly offended/complain about the lack of recognition by our/their peers. Feel free to get angry with me. I certainly welcome arguments and criticism.. As for me, I go back to find something useful in one of my textbooks. Laszlo Komuves FROM: L?szl? G. Komuves PhD Senior Principal Scientist, Manager, Microscopy Core Laboratory CORGENTECH, Inc. 650 Gateway Blvd., South San Francisco, CA 94080 Phone: (650) 246-6905, Fax: (650) 624-7540 E-mail: komuves@corgentech.com Date: Tue, 23 Nov 2004 08:02:24 -0500 From: Michele French Subject: [Histonet] IHC on Sections Processed for EM? To: histonet@lists.utsouthwestern.edu Message-ID: <41A334E0.2060308@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Good Morning Histonet! A colleague of mine did some EM work and found something interesting. Our pathologist wanted me to try to do some IHC to further characterize what is present in the section. Our EM person said it would never work. I did not think it was possible either, but I thought I would ask anyway. Has anyone tried doing an immunostain on plastic (Epon) sections from tissue fixed and processed for EM? I am always up for a challenge, but I am really busy right now and don't want to waste my time if there is really no hope! Thanks in advance, Michele French _______________________________________________ Histonet mailing list Histonet@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 james.zimmerman <@t> pharma.novartis.com Mon Nov 29 18:28:28 2004 From: james.zimmerman <@t> pharma.novartis.com (james.zimmerman@pharma.novartis.com) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] CD34 Message-ID: Hello, Does anyone have a procedure and or vendor for CD34 in Cyno monkeys in formalin fixed ,parrafin embedded tissues. Thanks, JPZ From Linresearch <@t> aol.com Mon Nov 29 18:42:45 2004 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Microarrayer Message-ID: <6a.497e6d96.2edd1c05@aol.com> Hello, Is there any recommendation for choosing a tissue microarrayer? Linda From mucram11 <@t> comcast.net Mon Nov 29 18:57:32 2004 From: mucram11 <@t> comcast.net (pam marcum) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] RE: IHC on tissues processed for EM References: <566FB0B522443D43AF02D2ADBE35A6F0010DF141@UTHEVS3.mail.uthouston.edu> Message-ID: <41ABC57C.000001.03944@YOUR-4DI1S53IME> Thank you, Barry. We all have days when even the simplest answers to common questions just aren't there and we ask the community of histologists we all belong to for help. Other times looking up the answer with the sources available can be best. We have to determine what we need and use those resources well. I think sometimes we use the easiest way to avoid taking the time to learn and everyone can be guilty of this at one time or another. It is only an issue when we take the easy way out every time. Often I see questions or requests for help when it is clear the person is in trouble, needs help and isn't even sure how to ask or what to ask. In that position they may not even know where to look as they are given a project and told do it with no help. Until we can all wiggle our noses and make it happen we should look at the community as place for help and not abuse it. I only have a problem when it is a student who clearly is asking us as a group to make it too easy for them to ask and not learn. The dangers of only knowing our field by rote or to repeat what they have read or heard is just not acceptable when the patient is at risk for not getting the best care. I feel the same way about research projects as the responsibility of the technician to make the effort to learn and grow everyday. Pam Marcum -------Original Message------- From: Rittman, Barry Date: 11/29/04 17:11:42 To: histonet Subject: RE: [Histonet] RE: IHC on tissues processed for EM Dawn et al. I do not think that this is a totally black and white problem. I think that it is entirely appropriate to ask people on Histonet for their opinion on a specific technique as there are many experts in the various fields. While I agree that there are no stupid questions, I believe that the intent of the writer was not to deride the individual but to suggest that they first try to search out the information before asking the question. I have taught students for many years and feel that more and more I am being asked to spoon feed individuals rather than to get them to learn the material. It is easier to provide the correct answer and considerably more difficult to motivate people to work out the solution for themselves (if they can) i.e. I believe that a ready made answer is usually not in the best interests of learning for that individual. When I was training (and I feel that I still am) while I could go and ask an "expert" next door for the answer, I always tried to find the answer out by myself first. This was, I believe, to my benefit as I was able not only to ask a more pointed question of the expert but also ask some important follow up questions. I believe that it is a much more satisfying approach. On the other side of the coin you have to realize that some of the individuals asking these questions may not have the resources available to them or even know where to begin to find out the information. I would suggest to those who wish to ask questions that you first try to research the material if you can as you will then be able to ask questions specific to your needs. You will benefit both ways. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of hymclab Sent: Monday, November 29, 2004 3:31 PM To: 'Gudrun Lang'; Histonetliste Subject: RE: [Histonet] RE: IHC on tissues processed for EM I too think that no question should be lableled as "easy". There are no stupid questions. I recently went through an issue about a stain that I have been doing for over 20 years. Couldn't figure out what was wrong. Technical services at two companies were stumped until someone came up with a possible answer. I tested it and they were correct. Someone else on the Histonetlist just had the same problem and I responded with what we found to be our problem. Then I got slammed by someone who couldn't have even read the whole message. I was answering someone'e problem about their mucicarmine stain not working. In my response to this person I used the terms Mayer's mucicarmine and Weigert's Hematoxylin. This other person that slammed accused me in living in the middle ages for using Mayer's Hematoxlyin and Weigert's hematoxlyin and said I should try Gill's hematoxylin. I didn't say I used Mayer's hematoxylin, I was referring to the name of the Mucicarmine stain. Secondly I had never heard of using Gill's in place of Weigert's in the Mucicarmine stain. I e-mailed him back I thought maybe he was confused---he said he wasn't and he said that he'd never even done a Mucicarmine stain. I rest my case. Things like that make you want to be a lurker instead of trying to help someone. Dawn -----Original Message----- From: Gudrun Lang [mailto:gu.lang@gmx.at] Sent: Thursday, November 25, 2004 3:09 AM To: Histonetliste Subject: Re: [Histonet] RE: IHC on tissues processed for EM Laszlo, I feel about saying some words about your message. You complain, that members of the histonetlist have no basic knowledgment. But what is basic? For me, in a routine surgery histolab, EM is very, very special for example. I belong to those people, that work in a small corner of the whole histological techniques. But I am interested and want to learn. You say, people have to use databases, text books etc. In my case I have no access through my lab to those things and they are'nt cheap (and first you have to know where to look). I am very happy about the histonetlist. Here are specialists, experts from every field and also "workmates". Please be tolerant with "easy" questions and problems, perhaps the person considers the problem more difficult than you know. friendly greetings Gudrun Lang ----- Original Message ----- From: "Laszlo Komuves" To: Sent: Wednesday, November 24, 2004 8:44 PM Subject: [Histonet] RE: IHC on tissues processed for EM There are hundreds, if not thousands of papers demonstrating successful localization of intracellular antigens at EM or LM level using tissue samples processed conventionally for EM (i.e. PFA/GA fixation, Os postfixation, epoxy resin embedding). Look for the now classical papers by Roth and Bendayan or their early reviews. Also the EM textbooks published in the 80's have plenty of information. And let me also add a personal comment of frustration: Histonet used to be about sharing experience and wisdom, mentoring and teaching those have were (are they still?) eager to learn or perfect their skills. I personally learnt/benefited from many messages posted. Now people are asking advice about coverslip sizes? So thanks to the curse of the Internet, journal articles and text books are used only by a dedicated few, electronic databases (libraries, PubMed, journal archives, reagent search sites, even Google) are not searched, and now even the vendor catalogs are not opened, because on the Histonet somebody will respond? How will users differentiate between solid information (based on scientific knowledge, practical experience) or just plain ignorance? Where is critical thinking? What happened to a once honorable guild of skilled craftsmanship, when members of the community are lacking elementary knowledge (just a few recent hair-raising topics: H&E staining, pH, and the list goes on and on)? And of course we/they are constantly offended/complain about the lack of recognition by our/their peers. Feel free to get angry with me. I certainly welcome arguments and criticism.. As for me, I go back to find something useful in one of my textbooks. Laszlo Komuves FROM: L?szl? G. Komuves PhD Senior Principal Scientist, Manager, Microscopy Core Laboratory CORGENTECH, Inc. 650 Gateway Blvd., South San Francisco, CA 94080 Phone: (650) 246-6905, Fax: (650) 624-7540 E-mail: komuves@corgentech.com Date: Tue, 23 Nov 2004 08:02:24 -0500 From: Michele French Subject: [Histonet] IHC on Sections Processed for EM? To: histonet@lists.utsouthwestern.edu Message-ID: <41A334E0.2060308@bms.com> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Good Morning Histonet! A colleague of mine did some EM work and found something interesting. Our pathologist wanted me to try to do some IHC to further characterize what is present in the section. Our EM person said it would never work. I did not think it was possible either, but I thought I would ask anyway. Has anyone tried doing an immunostain on plastic (Epon) sections from tissue fixed and processed for EM? I am always up for a challenge, but I am really busy right now and don't want to waste my time if there is really no hope! Thanks in advance, Michele French _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CrochiereSteve <@t> aol.com Mon Nov 29 20:24:01 2004 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Von kossa counterstain Message-ID: <1d9.30ecfd98.2edd33c1@aol.com> fred, In the past i've used nuclear fast red (kernechtrot) Steven M. Crochiere, HT(ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center Springfield, MA 01104 From dbpiontek <@t> clinomicsbio.com Mon Nov 29 20:56:45 2004 From: dbpiontek <@t> clinomicsbio.com (Denise Bland-Piontek) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Counterstain for Von Kossa Message-ID: <145F33B84793CF488578F86355B32E6912FCBD@mail.clinomicsbio.com> Just a note....... there are research companies that hire certified / trained staff. I have a staff of six that all take a great deal of pride in there work. Careful not to judge the masses by the actions of a few. I haven't been in this field as long as some, but I have several books and resources, one of which is the Histonet. There is, nonetheless, a reason why we have all subscribed (isn't there)? I have turned several "new techs" to the Histonet and their questions are significant to them. We were all there once upon a time and on our best day may still rely on the Histonet for help. I fear new techs are being driven away from a respectable community. Denise Bland-Piontek, HTL(ASCP)CTBS(AATB) Clinomics Biosciences, Inc. Histopathology Business Development Manager ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Paula Pierce Sent: Mon 11/29/2004 1:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Counterstain for Von Kossa Any Histology staining book will have a procedure for this stain. Lots of counterstains to choose from, but nuclear fast red is the standard. I am going to rant here, so please be patient with me. How many places out there are opening up labs or doing research with no trained people? And if these new people have internet access to find the histonet, they should be finding websites with actual procedures, AND most colleges and universities have libraries with REAL books on histology procedures. Go there and read. Since becoming registered 25 years ago, when I was asked to do something new I would go to my own library I have accumulated or the university library if it was really unusual and read, because we did not have the internet. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Tue Nov 30 02:34:41 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Water contamination [Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B545@bhrv-nt-11.bhrv.nwest.nhs.uk> I don't think fiddling with a few percentages of alcohol in the initial steps of dehydration makes any difference, bung it straight in 100%, it'll get diluted after a few batches. Change your last alcohol more regularly or have a 'drip bottle' of alcohol and, if you are manual staining, give the slides a quick wash of fresh alcohol before entering the first xylene. That's a point, did you say you were manual staining or not? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Jose Luis Palazon Fernandez [mailto:jluis.palazon@icman.csic.es] Sent: 29 November 2004 16:59 To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Water contamination [Scanned] In my lab we normally use 95 % EtOH before absolute alcohols, instead of the 70 % you use, without any problem. Maybe your contaminating agent is 70 % EtOH, try changing this step with a 95 % one Jos? Luis El dia 27/11/2004 19:32 usted envio el siguiente mensaje: >Date: 27 de Noviembre de 2004 19:32:07 >From: Paula Pierce >Subject: [Histonet] Water contamination >To: histonet@lists.utsouthwestern.edu > > First, you SLICE meat. Histotechs SECTION tissue. > > 70% EtOH is 30% water. You are probably getting too much water carryover into your 100%. Make sure the 100% EtOH is 100%. If the solutions are fresh, 10 to 20 dips each is enough time for dehydration before xylene and coverslipping. > > > 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 > Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Mandy <@t> serotec.co.uk Tue Nov 30 04:54:03 2004 From: Mandy <@t> serotec.co.uk (Mandy Townsend) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] CD34 Message-ID: Serotec are able to supply the anti CD34 antibody clone QBEND/10 (product code MCA547 or sample size MCA547T) that is suitable for use in paraffin sections and is cross reactive with cynomolgus monkeys. Please contact me or your local office, or visit the Serotec web site if you require further information. Mandy Mandy Townsend MSc Technical Services Advisor Serotec Ltd 22 Bankside Station Approach Kidlington Oxfordshire OX5 1JE Tel: +44 1865 852733 Fax: +44 1865 852739 email: mandy@serotec.co.uk URL: http://www.serotec.com/ Serotec-Your first choice for antibodies! Join our free email update service 4 antibodies for the price of 3 during October & November. More details IMPORTANT NOTICE: This message and any attachments may be confidential. If this has been sent to you in error, please contact the sender as soon as possible. Serotec Ltd. Registered in England No.1604642 Registered Office: Boswell House, 1-5 Broad Street, Oxford, OX1 SAW. UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of james.zimmerman@pharma.novartis.com Sent: Tuesday, November 30, 2004 12:28 AM To: Histonet@Pathology.swmed.edu Subject: [Histonet] CD34 Hello, Does anyone have a procedure and or vendor for CD34 in Cyno monkeys in formalin fixed ,parrafin embedded tissues. Thanks, JPZ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________ This e-mail has been scanned for all viruses by Star. The service is powered by MessageLabs. For more information on a proactive anti-virus service working around the clock, around the globe, visit: http://www.star.net.uk ________________________________________________________________________ From tony.j.savage <@t> gsk.com Tue Nov 30 05:16:33 2004 From: tony.j.savage <@t> gsk.com (tony.j.savage@gsk.com) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] H&E water contamination In-Reply-To: <200411291800.iATI0WWr000464@stvsfimr002.ggr.co.uk> Message-ID: Another source of water contamination in H&E sections can also occur in atmospheric conditions of high humidity (even in the UK), if there is the slightest delay in mounting the slides after their removal from xylene. This water contamination manifests itself as small collections of water micro droplets that seem to orientate themselves over nuclei, and look like a pigment lying over some of the nuclei. This artifact is also exhibits birefringence. It took me a while to track down and prove the source of this contamination. In the end I took slides out of xylene and breathed on them several times prior to mounting, and these slides showed the this same nuclear water artifact. The artifact is obviously caused by water vapour condensing on the slide prior to mounting but I cannot say what level of atmospheric humidity was required for this to manifest itself because I did not measure the humidity levels. Regards, Tony Histopathology Group Asthma Biology Department. RIRP CEDD. GlaxoSmithKline Medicines Research Centre, Gunnelswood Road, STEVENAGE, Hertfordshire. SG1 2NY tel. +44 (0)1438 764117 fax. +44 (0)1438 764782 email. Tony.J.Savage@gsk.com Message: 1 Date: Mon, 29 Nov 2004 10:00:02 +1100 From: Tony Henwood Subject: RE: [Histonet] H&E water contamination To: "'9msm8@qlink.queensu.ca'" <9msm8@qlink.queensu.ca>, histonet@lists.utsouthwestern.edu Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E123@simba.kids> Content-Type: text/plain A few ideas. Check your xylene for water. Keep the alcohol and xylene containers covered when not in use. Alcohol will absorb water from the air (possibly xylene as well - though of this I am not certain). 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: 9msm8@qlink.queensu.ca [mailto:9msm8@qlink.queensu.ca] Sent: Saturday, 27 November 2004 9:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] H&E water contamination Hi histonet I have been performing H&E stains on sliced parraffin embedded mouse hippocampal slices. I am consistently getting water contamination in my slides and have been unable to rectify the problem. My procedure for staining is: Xylene I for 7.5 minutes; Xylene II for 7.5 minutes; 100% ethanol I for 2 minutes; 100% ethanol II for 2 minutes; 95% ethanol I and 70% ethanol I for 2 minutes each; Water I for a second; then "H" for 20 minutes; Water II for 10 minutes; "E" for about 13 seconds, Water III for about 1 minute; 70% ethanol II for 50 seconds; 95% ethanol II for 5 minutes; 100% ethanol III and IV for 5 minutes each and then Xylene III and IV for 7.5 minutes each. I beleive the "70% ethanol II" is the key step for removing water after the staining but increasing the amount of time spent in the solution hasn't worked yet (though I have yet to leave it in for longer then a minute). Any tips would be greatly appreciated, Mike From jengirl1014 <@t> yahoo.com Tue Nov 30 06:52:04 2004 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Re: Counterstain for Von Kossa. Message-ID: <20041130125204.98288.qmail@web60601.mail.yahoo.com> I can give you the protocol that I have. Deparrafinize and rehydrate to dH2O. 5% Silver Nitrate solution, place in bright sunlight or in front of a 60 watt light bulb (place foil or mirror behind jar to reflect light. Leave for one hour or until calcium turns black. Rinse in dH2O, 3 changes.(I usually just use running dH2O for about 10-20 minutes) 5% NaThioSulfate for 5 min. Wash in tap H2O, rinse in distilled. Nuclear fast red for 5 minutes. Wash in water. Dehydrate and coverslip. Crystals will be black and cellular structures will be red/pink. If you have any more questions, feel free to write me. 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!? All your favorites on one personal page – Try My Yahoo! From ajennings <@t> unmc.edu Tue Nov 30 07:59:14 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Histolist helping "easy questions" In-Reply-To: <530361BF03351B4CAE5270A05D3037B503673600@bsrexms01.BSHSIR.COM> Message-ID: I never mind answering any question that I have the answer to. I enjoy sharing the knowledge I have and will never claim to be the definitive answer, so I encourage all to use whatever resources are available to them. I have to admit some of us don't realize that computer access is not always available for searching....example I told one of my techs that if she couldn't get in touch with me by phone to please email me that she couldn't come into work......her reply was....I would have to come in to email you that I couldn't come in because I don't have a computer at home. bottom line....I just wanted to write on this topic so I could change the subject line because there were more non topic replies than replies to the original post :-) anita From RMaliniak <@t> SBHCS.com Tue Nov 30 09:16:08 2004 From: RMaliniak <@t> SBHCS.com (Maliniak, Richard) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Autoimmunostainers DAKO vs Ventanna Message-ID: <69223CA47975154E90C986BC075F1EB3FAFE62@SBCEXCHANGE01.sbhcs.com> I am looking for info Ventanna to DAKO and DAKO to V did you like or dislike abo Rick ________________________________________________________________ Important news about our email communications Saint Barnabas Health Care System has implemented secure messaging servic To learn more about SBHCS Secure Messaging, go to: [1]http://www.zixcorp.c If you need assistance with retrieving a secure email, please email sbhcs [2]http://www.zixcorp.com/evangelism/sbhcs/par References 1. 3D"http://www.zixcorp.com/evangelism/sbhcs/" 2. 3D"http://www.zixcorp.com/evangelism/ From twebster <@t> nmcinc.org Tue Nov 30 09:30:23 2004 From: twebster <@t> nmcinc.org (Tim Webster) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Ammonia water pH Message-ID: 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. > > From cgfields <@t> lexhealth.org Tue Nov 30 09:32:46 2004 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Cryostat Repair Message-ID: We use.... Southeast Pathology Instrument Service Phone 843-588-2559 cell-843-708-7090 Michael Dietrich is the person that does the repairs. He does a very good job and is trained for Sakura, Lieca, Microm and most any equipment. He has worked on our cryostats for several years and the new ones I purchased. Good luck Carole Fields Lex. Med Ctn. W.Columbia, SC -----Original Message----- From: Kevin Briggs [mailto:BriggsK@drmc.drhsi.org] Sent: Wednesday, November 24, 2004 12:14 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cryostat Repair Hello, does anyone know of a good repair center or person in Virginia or North Carolina who services cryostats? Thanks, Kevin D. Briggs, MS, CT(ASCP) Team Leader- Cytopathology/ Histopathology Services Danville Regional Medical Center 142 South Main Street Danville, VA 24541 Telephone: (434) 799-4470 ext.5451 Fax: (434) 799-2118 E-mail: briggsk@drhsi.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From Kemlo.Rogerson <@t> elht.nhs.uk Tue Nov 30 09:36:03 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B549@bhrv-nt-11.bhrv.nwest.nhs.uk> 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 From Jackie.O'Connor <@t> abbott.com Tue Nov 30 09:35:55 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Ammonia water pH Message-ID: 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 From Kemlo.Rogerson <@t> elht.nhs.uk Tue Nov 30 10:08:53 2004 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Ammonia water pH[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D01B0B54A@bhrv-nt-11.bhrv.nwest.nhs.uk> Bang on! Weak is 0.2 to 0.3% NH40H, anymore and the sections disappear. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: 30 November 2004 15:36 To: Tim Webster Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Ammonia water pH[Scanned] 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 From Rcartun <@t> harthosp.org Tue Nov 30 10:12:28 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods? Message-ID: 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 From gcallis <@t> montana.edu Tue Nov 30 10:59:12 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Re: Ammonia water pH and other bluing solutions In-Reply-To: References: Message-ID: <6.0.0.22.1.20041130094056.01b3d3e0@gemini.msu.montana.edu> You are correct in your concern about losing sections/wrinkling. Although we use a commercial solution, the ideal pH is 8. We no longer use dilute ammonia water, as with higher pH, it is likely the alkaline conditions can cause section to fall off the slide. (hmm isn't this called alkaline protein hydrolysis that affects alkaline sensitive protein bonds. Been there and had it happen particularly with large, precious difficult to obtain decalcified bone sections. We change the bluing solution DAILY in order to maintain our pH at 8. Since water carryover can dilute the bluing solution further after many slides are run through, the first of the week fresh change may have correct pH but end of week may be entirely different after the watercarry over into bluing solution. Bluing solution is cheap. This also goes for dilute acetic acid solution after hematoxylin., particularly progressive hematoxylins such as Gills, Richard Allen Hematoxylin 1, etc. An excellent weak alkaline solution is Scotts tap water substitute made with salts that are more alkaline in nature. You can make this up in your lab just as easily as ammonia water, but is is much gentler to the section. Daily changes would not be a huge expense. It is also available as a concentrate from various sources, Sigma and Surgipath come to mind. Never lost a section when using this reagent, but ammonia water has caused grief in the past. 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 Jackie.O'Connor <@t> abbott.com Tue Nov 30 11:14:25 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods? Message-ID: 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 From contact <@t> excaliburpathology.com Tue Nov 30 12:17:12 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Ammonia water pH Message-ID: <20041130181712.32329.qmail@web50303.mail.yahoo.com> 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 From susan.wells <@t> bms.com Tue Nov 30 12:21:51 2004 From: susan.wells <@t> bms.com (Susan Q Wells) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Frozen cell pellet Message-ID: <41ACBA3F.9030602@bms.com> 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 From cfavara <@t> niaid.nih.gov Tue Nov 30 12:38:40 2004 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Frozen cell pellet Message-ID: 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 From TJJ <@t> Stowers-Institute.org Tue Nov 30 12:51:22 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Re: Ammonia water pH and other bluing solutions Message-ID: 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 From BMolinari <@t> heart.thi.tmc.edu Tue Nov 30 13:07:43 2004 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] gliut/isopropyl alcohol Message-ID: 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 From Jackie.O'Connor <@t> abbott.com Tue Nov 30 13:08:41 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Re: Ammonia water pH and other bluing solutions Message-ID: 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 From vazquezr <@t> ohsu.edu Tue Nov 30 13:38:08 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Re: Ammonia water pH and other bluing solutions Message-ID: 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 From darrenj <@t> medica.co.nz Tue Nov 30 13:49:29 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] In-Reply-To: <1030B679AD69D6119C3F00080210DD9D01B0B549@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: <000001c4d715$b49168d0$c864a8c0@medica.co.nz> 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 From gcallis <@t> montana.edu Tue Nov 30 13:45:41 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Frozen cell pellet In-Reply-To: <41ACBA3F.9030602@bms.com> References: <41ACBA3F.9030602@bms.com> Message-ID: <6.0.0.22.1.20041130123759.01b57008@gemini.msu.montana.edu> 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) From gcallis <@t> montana.edu Tue Nov 30 13:58:26 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Saturated lithium carbonate bluing In-Reply-To: References: Message-ID: <6.0.0.22.1.20041130124819.01b66e58@gemini.msu.montana.edu> 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) From darrenj <@t> medica.co.nz Tue Nov 30 14:28:05 2004 From: darrenj <@t> medica.co.nz (Darren James) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Autoimmunostainers DAKO vs Ventanna[Scanned] In-Reply-To: <000001c4d717$4ed973f0$3601a8c0@brownpathology.net> Message-ID: <000801c4d71b$198aeb80$c864a8c0@medica.co.nz> 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 From TJJ <@t> Stowers-Institute.org Tue Nov 30 15:09:05 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Re: Ammonia water pH and other bluing solutions Message-ID: 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 From TJJ <@t> Stowers-Institute.org Tue Nov 30 15:13:43 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Paraffin processing explant cultures Message-ID: 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 From gu.lang <@t> gmx.at Tue Nov 30 15:59:33 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:20 2005 Subject: [Histonet] Re: Ammonia water pH and other bluing solutions References: Message-ID: <003a01c4d727$e077a3d0$eeeea8c0@server> 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 > From LuckG <@t> empirehealth.org Tue Nov 30 15:59:19 2004 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods? Message-ID: 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 From fredrik <@t> u.washington.edu Tue Nov 30 15:59:42 2004 From: fredrik <@t> u.washington.edu (Fredrik Johansson) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Counterstain for Von Kossa Message-ID: <239E252E-431B-11D9-94FE-000A95F0ED90@u.washington.edu> 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 From MinHan.Tan <@t> vai.org Tue Nov 30 16:02:35 2004 From: MinHan.Tan <@t> vai.org (Tan, MinHan) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Phosphorylated-met antibodies Message-ID: 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. From liz <@t> premierlab.com Tue Nov 30 16:02:30 2004 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin processing explant cultures Message-ID: <000001c4d728$49c9c480$76d48a80@AMY> 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 From nperson211 <@t> comcast.net Tue Nov 30 16:24:50 2004 From: nperson211 <@t> comcast.net (nperson211@comcast.net) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] CD 133 Message-ID: <113020042224.12979.41ACF331000E7125000032B32207001641CECECD02019C9D0A9F02@comcast.net> 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 From TJJ <@t> Stowers-Institute.org Tue Nov 30 17:01:52 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin processing explant cultures Message-ID: 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 From vazquezr <@t> ohsu.edu Tue Nov 30 17:29:32 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods? Message-ID: 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 From rockbeki <@t> ufl.edu Tue Nov 30 18:13:50 2004 From: rockbeki <@t> ufl.edu (SMITH,REBEKAH FELICIA) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Ammonia water pH Message-ID: <934672373.1101860030245.JavaMail.osg@osgjas02.cns.ufl.edu> 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" From JMacDonald <@t> mtsac.edu Tue Nov 30 18:17:46 2004 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Paraffin blocks - Dangerous goods? In-Reply-To: Message-ID: 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 From RCHIOVETTI <@t> aol.com Tue Nov 30 18:26:06 2004 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] gliut/isopropyl alcohol Message-ID: <1cc.2d22ffb1.2ede699e@aol.com> 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 jwatson <@t> gnf.org Tue Nov 30 19:28:20 2004 From: jwatson <@t> gnf.org (James Watson) Date: Fri Sep 16 15:24:21 2005 Subject: [Histonet] Part time Histology position in San Diego Ca. Message-ID: 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