From louise.marks <@t> syngenta.com Wed Sep 1 05:33:42 2004 From: louise.marks <@t> syngenta.com (louise.marks@syngenta.com) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] drying out of sections Message-ID: <7D28784DD2EBD511AC4900508B6219C103F5F948@ukapagmbx01.ukct.zeneca.com> Dear All, I am having problems finding a suitable mountant to use on quite thick (40 micron) free floating stained sections mounted onto slides. The mountant I currently use, DPX, appears to be OK for the first few months but then sections appear to 'dry out'- air forms round the edge of the section then moves over the surface. I am after some advice on a good mountant to use on sections for long term storage and any other tips to prevent this drying out..best storage conditions etc. I use the slides for stereology at 100x magnification so I would need a mountant that would be compatible with microsopy, Any tips / info much appreciated, Regards Louise Dr Louise Marks Syngenta CTL UK From David.Edmondson <@t> christie-tr.nwest.nhs.uk Wed Sep 1 06:28:12 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Xam and Eosin Message-ID: Hi folks, Yesterday I was phoning in an order with VWR and find that they can no longer supply Xam or Spirit soluble Eosin any more. Do you know of other suppliers ? Sigma and VWR have DPX Is the problem international? Hoarding is not on the agenda. David Christie Hospital Manchester UK From lpwenk <@t> sbcglobal.net Wed Sep 1 07:11:03 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] registry tissue Message-ID: <004601c4901c$c2297f40$ea36d445@domainnotset.invalid> To those writing in asking for help getting their registry tissue - please include additional information about the size and content of the tissue. This information was included in your packet, and the rest of the Histonet community does not have this information (And be aware that the sizes listed are AFTER processing, so if tissue in fixative is sent then it must be larger.) For example: - Artery that was requested (HT or HTL) must be 0.5 cm in diameter and a complete cross-section. So any size will not do, and a longitudinal will not do. - Lung - HT needs lung that is 1 x 1 cm square, while HTL needs lung that is 1.5 x 1.5 cm square and must have cartilage/bronchus in it. - Uterus for HT or HTL must be 1.5 x 1.5 cm square, with myometrium and endometrium, and the endometrium must cover one entire surface. This means the uterus had to be big to begin with, not from a post-menopausal 80 year old. - Small intestine for both HT and HTL must be 2 cm in length, contain all layers, not be autolyzed, and the epithelium must cover 1 entire surface. - Liver for both HT and HTL needs to be 1 x 1 cm square. No other criteria. I know these criteria, because I help my HT and my HTL students get their tissues every year. Most people on Histonet would not know these criteria. These criteria are VERY important to know, before someone volunteers to help collect tissue. If these criteria are not followed, points will be deducted, and the person doing the exam will in most likelihood fail. Plus, it has been a waste of valuable time for the kind person trying to obtain the tissue. Please help us to help you. Peggy A. Wenk, HTL(ASCP)SLS Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 From mark.lewis <@t> thermo.com Wed Sep 1 08:04:26 2004 From: mark.lewis <@t> thermo.com (mark.lewis@thermo.com) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Equipment Validation Message-ID: Good morning everyone. I have a question concerning equipment validation. We recently had a customer that said they were cited during an inspection for not having a record of their new equipment being validated. I am assume that the validation of the equipment means that they should have run a parallel test with the new and "old" piece of equipment to verify that it indeed performs as it should. The customer felt it was our responsibility to do the validation testing in their lab for the equipment. Does anyone have some more insight or protocols as to what equipment validation documentation is required of inspecting agencies like Joint Commission ? I appreciate your input. Have a Blessed Day ! Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com From BSylinda <@t> aol.com Wed Sep 1 08:05:13 2004 From: BSylinda <@t> aol.com (BSylinda@aol.com) Date: Fri Sep 16 15:23:58 2005 Subject: [Histonet] Updated tissue for registry Message-ID: <324608D8.2821B119.00698496@aol.com> Hello, I am writing again, hoping that someone may have access to some of this tissue: artery- complete cross section (0.5 cm outside diameter) liver- 1.0 x 1.0 cm square lung- 1.0 x 1.0 cm square uterus- 1.5 x 1.5 cm square -to include endometrium and myometrium -endometrium to cover one entire surface small intestine- 2.0 cm in length -not autolyzed -to include all layers -epithelium to cover one entire surface Thanks, Sylinda Battle Pathology Services 1002 Tx. Blvd. suite 500 Texarkana, TX 75501 From Joyce.Rush <@t> sjmcmn.org Wed Sep 1 08:36:16 2004 From: Joyce.Rush <@t> sjmcmn.org (Rush, Joyce) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Equipment Validation Message-ID: <494BA9CA5AA74C48B644FCA2B8BECEC844D9AF@sjw3smail.SJMCMN.ORG> Mark, In the clinical lab our expectation is that the instrument vendor will perform the validation on our site. They run our specimens on their machine using reagents at no cost to us and crunch the data. For a processor it was not our expectation that the vendor run the parallels since the pathologists judge the final outcome. We compared every special stain, immuno, etc on tissue run on both old and new processors. I guess we could have required that the vendor pay for reagents for the crossover but I guess we did not think of it, darn it! It surely is painful but required as far as I know. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of mark.lewis@thermo.com Sent: Wednesday, September 01, 2004 08:04 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Equipment Validation Good morning everyone. I have a question concerning equipment validation. We recently had a customer that said they were cited during an inspection for not having a record of their new equipment being validated. I am assume that the validation of the equipment means that they should have run a parallel test with the new and "old" piece of equipment to verify that it indeed performs as it should. The customer felt it was our responsibility to do the validation testing in their lab for the equipment. Does anyone have some more insight or protocols as to what equipment validation documentation is required of inspecting agencies like Joint Commission ? I appreciate your input. Have a Blessed Day ! Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: 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 sender thereof and destroy/delete the message. Please note that any views or opinions presented in this email are solely those of the Author and do not necessarily represent those of the company. Finally, the recipient should check this email and any attachments for the presence of viruses. The Medical Center accepts no liability for any damage caused by any virus transmitted by this email. St. Joseph's Medical Center, 523 N. 3rd street, Brainerd, MN 56401 From garygill <@t> dcla.com Wed Sep 1 08:58:15 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] drying out of sections Message-ID: <0B11620AEE5EFB4FA7A7558339972A8307058F@HALIBUT.dcla.com> Thick sections challenge mounting media to remain intact, without retraction (i.e., air under the cover glass), because the thick edge seal along the perimeter allows solvent to evaporate more rapidly than it would with thin sections. The plastic resin cannot form a barrier, so air creeps in. There are three obvious potential solutions: 1) Change to a mounting medium that has a lower proportion of solute. Unfortunately, without doing weight loss experiments on your own, you probably won't get the desired weight/volume information from the label on the bottle of mounting medium for comparison. You can always try asking the vendor for the information. Whoever makes it knows the answer. 2) Use less mounting medium. 3) "Cook" the covered slides. Cooking slides originated in the lab of the late Dr. Ruth Graham in the 1950s. Freshly covered slides are set on a hot plate, covered with aluminum foil for easy subsequent clean-up, set to a temperature slightly above the boiling point of xylene (the usual solvent). As I vaguely recall, that's about 117 degree C. As soon as the mounting medium begins to boil in a few seconds, remove the slide and set it on a work bench. Use forceps to tap out the bubbles, square the cover glass on the slide, and clean with xylene. It's messy, but it solidifies the mounting medium immediately, reduces its thickness so microscopic examination with high numerical aperture objectives is optimized, and promotes long-term dye stability. Gary Gill -----Original Message----- From: louise.marks@syngenta.com [mailto:louise.marks@syngenta.com] Sent: Wednesday, September 01, 2004 5:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] drying out of sections Dear All, I am having problems finding a suitable mountant to use on quite thick (40 micron) free floating stained sections mounted onto slides. The mountant I currently use, DPX, appears to be OK for the first few months but then sections appear to 'dry out'- air forms round the edge of the section then moves over the surface. I am after some advice on a good mountant to use on sections for long term storage and any other tips to prevent this drying out..best storage conditions etc. I use the slides for stereology at 100x magnification so I would need a mountant that would be compatible with microsopy, Any tips / info much appreciated, Regards Louise Dr Louise Marks Syngenta CTL UK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Wed Sep 1 09:03:06 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] re Muscle regeneration Message-ID: <003c01c4902c$6799f400$83509b51@home> CD56 ( NCAM) is highly upregulated, if I recall , in regen fibres. --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.726 / Virus Database: 481 - Release Date: 22/07/2004 From mprice26 <@t> juno.com Wed Sep 1 09:25:17 2004 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] RE: Biocare's Info Message-ID: <20040901.072523.1142.491671@webmail26.nyc.untd.com> I need the contact information for Biocare Medical. Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From pruegg <@t> ihctech.net Wed Sep 1 09:45:11 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Updated tissue for registry In-Reply-To: <324608D8.2821B119.00698496@aol.com> Message-ID: Sylinda, NSH maintains a tissue bank but they are paraffin blocks we do not have wet tissue which is what I think you will need for the HT exam as you are suppose to process your own. Contact coroner's or University in your area and ask for autopsy tissue. If you do autopsy at your place ask your morgue deaner or the pathologist for help with this. You can also try to access surgical specimens from where ever they go to be grossed. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of BSylinda@aol.com Sent: Wednesday, September 01, 2004 6:05 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Updated tissue for registry Hello, I am writing again, hoping that someone may have access to some of this tissue: artery- complete cross section (0.5 cm outside diameter) liver- 1.0 x 1.0 cm square lung- 1.0 x 1.0 cm square uterus- 1.5 x 1.5 cm square -to include endometrium and myometrium -endometrium to cover one entire surface small intestine- 2.0 cm in length -not autolyzed -to include all layers -epithelium to cover one entire surface Thanks, Sylinda Battle Pathology Services 1002 Tx. Blvd. suite 500 Texarkana, TX 75501 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Wed Sep 1 09:45:08 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] NSH S/C in Toronto Message-ID: I am writing to invite those attending the NSH S/C in Toronto to attend the IHC Resource Group committee meeting held Sat. Sept. 18 at 5:30 PM. We have some exciting IHC issues on the agenda to discuss and we are asking for agenda items now to be included. If there is an IHC issue you would like to have discussed at this meeting, let me know and I will include it on the agenda. If you are not a member of this IHC Resource Group and would like to be, go to www.ihcrg.org and register online. This is a committee of NSH and membership with NSH is required. Patsy Ruegg, Chair NSH IHCRG From TJJ <@t> Stowers-Institute.org Wed Sep 1 09:49:38 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] CD4 and CD8 in paraffin? Message-ID: In this latest issue of Applied Immunohistochemistry and Molecular Morphology (AIMM 2004; 12:198-204) is an article in which CD4 and CD8 are used in paraffin sections with antigen unmasking. CD4 (clone 1F6, 1:20 from Novocastra, Malvern, Australia) and CD8 (clone C8/144B, 1:200; Dako, Sydney, Australia) were done on routinely processed duodenal biopsies. Maybe we're in the wrong hemisphere and that explains why it doesn't work for us? Cheers! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From akbitting <@t> geisinger.edu Wed Sep 1 10:00:46 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Factor XIIIa Message-ID: Thanks for all the replys to my query about finding a new supplier for Factor XIIIa. I called DAKO and they told me that they DO NOT have Factor XIIIa. Someone yesterday emailed me and said they have a polyclonal Factor XIIIa that they think works very well. If youre out there, email me again with the product info please! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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 juan.gutierrez <@t> christushealth.org Wed Sep 1 10:01:40 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] CD4 and CD8 in paraffin? Message-ID: We run them all the time on the Benchmarks. Ventana sells the primaries, probably from NCL, and we've gotten excellent results on GI bx. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My views are my own and do not reflect those of my employer. Long live free speech! "We have nothing to fear but fear itself". -----Original Message----- From: Johnson, Teri [mailto:TJJ@Stowers-Institute.org] Sent: Wednesday, September 01, 2004 9:50 AM To: Histonet Subject: [Histonet] CD4 and CD8 in paraffin? In this latest issue of Applied Immunohistochemistry and Molecular Morphology (AIMM 2004; 12:198-204) is an article in which CD4 and CD8 are used in paraffin sections with antigen unmasking. CD4 (clone 1F6, 1:20 from Novocastra, Malvern, Australia) and CD8 (clone C8/144B, 1:200; Dako, Sydney, Australia) were done on routinely processed duodenal biopsies. Maybe we're in the wrong hemisphere and that explains why it doesn't work for us? Cheers! 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 lucyb <@t> biocare.net Wed Sep 1 10:23:11 2004 From: lucyb <@t> biocare.net (Lucy Brooks) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] I have a funny References: Message-ID: <003201c49037$9950d8f0$0a01a8c0@LUCYSALES> I personally hope the histonet isn't used to "intimidate companies" , but I do feel that if you have a problem and the company who you have that problem with doesn't deal with it in a reasonable amount of time, it is nice to warn other people out their of what they can expect when dealing with these said companies. ----- Original Message ----- From: "Sarah Jones" To: ; ; Sent: Tuesday, August 31, 2004 6:56 PM Subject: Re: [Histonet] I have a funny > Ah, this begs an ethical question.... should the histonet be used as a > threat to intimidate companies? > Is this an Us against Them mentality? > Discuss amongst yourselves. (a little "Coffee Talk" humor there for > you Saturday Night Live fans) > Oye Vey..................... Sarah > > > Sarah Jones, HT(ASCP) > Histology Lab > Dept. of Veterinary Integrative Biosciences > College of Veterinary Medicine > Texas A&M University > College Station, TX 77843-4458 > phone: 979-845-3177 > fax: 979-458-3499 > > > >>> "Lucy Brooks" 8/31/2004 6:10:52 PM >>> > Joe!! > > Speak your mind!!! > The Histonet is a powerful thing!! > > ----- Original Message ----- > From: "Joe Nocito" > To: "Histonet" > Sent: Tuesday, August 31, 2004 4:01 PM > Subject: [Histonet] I have a funny > > > > Hello all, > > for once, I have, what I think is funny, is a short story. A couple > of > > months ago, I purchased a piece of equipment from a vendor (who will > remain > > anonymous, at this time at least). I was told it was 6 weeks for > delivery. > > It's been over 8 weeks. I called the rep and left a voice mail asking > when > I > > can expect my equipment. S/he called me right back. I asked when I > can > > expect the equipment. Then I said "Don't make me go on the > Histonet". > > My techs started laughing and I couldn't keep a straight face. The > rep was > > all apologetic and promised me I'll have my equipment in 2 weeks. > Gee, I > > didn't think I was getting that bad. Maybe I should settle down a > little > > bit. > > > > 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 cjohnston <@t> mdanderson.org Wed Sep 1 11:05:36 2004 From: cjohnston <@t> mdanderson.org (cjohnston@mdanderson.org) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] 5-Fam Message-ID: Hi All, I am looking for information on 5-Fam = 5 Carboxyflouresceine. This is to be attached to a peptide and then cut and stained. Thank you Carol Carol M. Johnston HT(ASCP) M.D. Anderson Cancer Center 1515 Holcombe Blvd. Unit 443 Houston, Texas 77030 713-745-4625 From kappeler <@t> patho.unibe.ch Wed Sep 1 11:05:44 2004 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Factor XIIIa References: Message-ID: <00d301c4903d$896f15e0$27955c82@patho.unibe.ch> Angela, once you have sorted out all those that have difficulties in making the difference between VIII (8, eight) and XIII (13, thirteen) - could you let us know who finally was able to supply the antibody? Thanks a lot! Andi Kappeler Institute of Pathology, University of Bern, Switzerland ----- Original Message ----- From: "Angela Bitting" To: Sent: Wednesday, September 01, 2004 5:00 PM Subject: [Histonet] Factor XIIIa > Thanks for all the replys to my query about finding a new supplier for Factor XIIIa. I called DAKO and they told me that they DO NOT have Factor XIIIa. > Someone yesterday emailed me and said they have a polyclonal Factor XIIIa that they think works very well. If youre out there, email me again with the product info please! > > Angela Bitting, HT(ASCP) > Technical Specialist, Histology > Geisinger Medical Center > 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 mark.lewis <@t> thermo.com Wed Sep 1 11:10:09 2004 From: mark.lewis <@t> thermo.com (mark.lewis@thermo.com) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Equipment Validation Message-ID: Joyce, Thanks for your reply. I think it is a good idea for the vendor to supply start-up reagents. I was wondering if you had a copy of the specific questions that the inspectors ask for with regard to validation. If so could you fax or e-mail a copy to me? Sure does sound like a lot of work especially with regards to immuno staining. Thank You Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation Phone (412) 747-4013 Fax (412) 788-6557 E-mail: mark.lewis@thermo.com "Rush, Joyce" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] Equipment Validation western.edu 09/01/2004 09:36 AM Mark, In the clinical lab our expectation is that the instrument vendor will perform the validation on our site. They run our specimens on their machine using reagents at no cost to us and crunch the data. For a processor it was not our expectation that the vendor run the parallels since the pathologists judge the final outcome. We compared every special stain, immuno, etc on tissue run on both old and new processors. I guess we could have required that the vendor pay for reagents for the crossover but I guess we did not think of it, darn it! It surely is painful but required as far as I know. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of mark.lewis@thermo.com Sent: Wednesday, September 01, 2004 08:04 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Equipment Validation Good morning everyone. I have a question concerning equipment validation. We recently had a customer that said they were cited during an inspection for not having a record of their new equipment being validated. I am assume that the validation of the equipment means that they should have run a parallel test with the new and "old" piece of equipment to verify that it indeed performs as it should. The customer felt it was our responsibility to do the validation testing in their lab for the equipment. Does anyone have some more insight or protocols as to what equipment validation documentation is required of inspecting agencies like Joint Commission ? I appreciate your input. Have a Blessed Day ! Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: 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 sender thereof and destroy/delete the message. Please note that any views or opinions presented in this email are solely those of the Author and do not necessarily represent those of the company. Finally, the recipient should check this email and any attachments for the presence of viruses. The Medical Center accepts no liability for any damage caused by any virus transmitted by this email. St. Joseph's Medical Center, 523 N. 3rd street, Brainerd, MN 56401 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Wed Sep 1 11:09:47 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] RE: Biocare's Info Message-ID: Biocare's phone #: (800)799-9499. 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: mprice26@juno.com [mailto:mprice26@juno.com] Sent: Wednesday, September 01, 2004 9:25 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Biocare's Info I need the contact information for Biocare Medical. Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Sep 1 11:32:52 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] CD4 and CD8 in paraffin? In-Reply-To: <200409011457.i81EvTmo022224@mail1.msu.montana.edu> References: <200409011457.i81EvTmo022224@mail1.msu.montana.edu> Message-ID: <6.0.0.22.1.20040901102833.01b05090@gemini.msu.montana.edu> Teri et al, You did not mention the species. If the staining was on human duodenal biopsies (tough thing to do on a mouse!) rather than mouse tissue then these markers work. The Mouse remains a total bust! Boo Hoo! Must be the critter aka species differences rather than living Down Under, eh!?? At 08:49 AM 9/1/2004, you wrote: >In this latest issue of Applied Immunohistochemistry and Molecular >Morphology (AIMM 2004; 12:198-204) is an article in which CD4 and CD8 >are used in paraffin sections with antigen unmasking. CD4 (clone 1F6, >1:20 from Novocastra, Malvern, Australia) and CD8 (clone C8/144B, 1:200; >Dako, Sydney, Australia) were done on routinely processed duodenal >biopsies. Maybe we're in the wrong hemisphere and that explains why it >doesn't work for us? > >Cheers! > > >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 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 juan.gutierrez <@t> christushealth.org Wed Sep 1 11:54:33 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] I have a funny Message-ID: Been there, done that, and got my a.. chewed out by my boss when they threatened legal action. Maybe if those companies spent less time on the histonet and more time taking care of the customer this thread will go away. NOW I DID NOT MENTION ANY NAMES SO DON'T CALL ME OR MY BOSS, OR YOUR LAWYERS, OR MY LAWYERS, OR YOUR LAWYERS' LAWYER, OR YOUR COUSIN THE LAWYER. ALSO DON'T HAVE YOUR PEOPLE CALL MY PEOPLE EITHER, I DON'T WANT TO DO LUNCH! Of course I'm referring to the companies and not to you Lucy. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My views are my own and do not reflect those of my employer. Long live free speech! "We have nothing to fear but fear itself". -----Original Message----- From: Lucy Brooks [mailto:lucyb@biocare.net] Sent: Wednesday, September 01, 2004 10:23 AM To: Sarah Jones; histonet@pathology.swmed.edu; JNocito@Pathreflab.com Subject: Re: [Histonet] I have a funny I personally hope the histonet isn't used to "intimidate companies" , but I do feel that if you have a problem and the company who you have that problem with doesn't deal with it in a reasonable amount of time, it is nice to warn other people out their of what they can expect when dealing with these said companies. ----- Original Message ----- From: "Sarah Jones" To: ; ; Sent: Tuesday, August 31, 2004 6:56 PM Subject: Re: [Histonet] I have a funny > Ah, this begs an ethical question.... should the histonet be used as a > threat to intimidate companies? > Is this an Us against Them mentality? > Discuss amongst yourselves. (a little "Coffee Talk" humor there for > you Saturday Night Live fans) > Oye Vey..................... Sarah > > > Sarah Jones, HT(ASCP) > Histology Lab > Dept. of Veterinary Integrative Biosciences > College of Veterinary Medicine > Texas A&M University > College Station, TX 77843-4458 > phone: 979-845-3177 > fax: 979-458-3499 > > > >>> "Lucy Brooks" 8/31/2004 6:10:52 PM >>> > Joe!! > > Speak your mind!!! > The Histonet is a powerful thing!! > > ----- Original Message ----- > From: "Joe Nocito" > To: "Histonet" > Sent: Tuesday, August 31, 2004 4:01 PM > Subject: [Histonet] I have a funny > > > > Hello all, > > for once, I have, what I think is funny, is a short story. A couple > of > > months ago, I purchased a piece of equipment from a vendor (who will > remain > > anonymous, at this time at least). I was told it was 6 weeks for > delivery. > > It's been over 8 weeks. I called the rep and left a voice mail asking > when > I > > can expect my equipment. S/he called me right back. I asked when I > can > > expect the equipment. Then I said "Don't make me go on the > Histonet". > > My techs started laughing and I couldn't keep a straight face. The > rep was > > all apologetic and promised me I'll have my equipment in 2 weeks. > Gee, I > > didn't think I was getting that bad. Maybe I should settle down a > little > > bit. > > > > 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Wed Sep 1 12:00:24 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Factor XIIIa Message-ID: Cell Marque has both. I have not used them, but I have used some of their other products with good results. They'll even send you a sample. 1-800-665-7284 Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My views are my own and do not reflect those of my employer. Long live free speech! "We have nothing to fear but fear itself". -----Original Message----- From: Andi Kappeler [mailto:kappeler@patho.unibe.ch] Sent: Wednesday, September 01, 2004 11:06 AM To: Angela Bitting; Histonet Subject: Re: [Histonet] Factor XIIIa Angela, once you have sorted out all those that have difficulties in making the difference between VIII (8, eight) and XIII (13, thirteen) - could you let us know who finally was able to supply the antibody? Thanks a lot! Andi Kappeler Institute of Pathology, University of Bern, Switzerland ----- Original Message ----- From: "Angela Bitting" To: Sent: Wednesday, September 01, 2004 5:00 PM Subject: [Histonet] Factor XIIIa > Thanks for all the replys to my query about finding a new supplier for Factor XIIIa. I called DAKO and they told me that they DO NOT have Factor XIIIa. > Someone yesterday emailed me and said they have a polyclonal Factor XIIIa that they think works very well. If youre out there, email me again with the product info please! > > Angela Bitting, HT(ASCP) > Technical Specialist, Histology > Geisinger Medical Center > 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 > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From daniel.eberhard <@t> uni-bielefeld.de Wed Sep 1 12:00:04 2004 From: daniel.eberhard <@t> uni-bielefeld.de (DANIEL EBERHARD) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] re-staining of "exhumed" sections Message-ID: <41360014.2070800@UNI-BIELEFELD.DE> Dear Histonetters, I would like to re-stain old pFA fixed sections, which developed high auto-fluorescence levels after some month. Thus, I decoversliped them, removed (washed) the mounting media and performed normal antibody staining using sec HRP conj. AB and DAB. Unfortunately, some AB which recognize their epitopes in "fresh" pFA fixed sections do not work on the "exhumed" sections. I wonder if someone can assist me in solving this problem. Might antigen retrieval methods help in this case? Any hints may be helpful, thanks in advance Daniel From amosbrooks <@t> earthlink.net Wed Sep 1 12:03:27 2004 From: amosbrooks <@t> earthlink.net (amosbrooks@earthlink.net) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] equipment validation Message-ID: <30583862.1094058207640.JavaMail.root@ernie.psp.pas.earthlink.net> Hi, Just to be on the safe side it would be wise if you knew what equipment requires validation and advise your customers that it needs to be done. Some customers may be perfectly capable and willing to do it themselves. Others will probably expect you to do it for them. This puts you in a good position to perform the validation but charge for the service. The catch is that inspections and validations are somewhat subjective. Make sure that if you do a validation that it is rock solid for an inspection. I really see no reason why a company should be expected to do this for all situations though. Just make sure the customer knows that it needs to be done. Amos Brooks ============================================= Good morning everyone. I have a question concerning equipment validation. We recently had a customer that said they were cited during an inspection for not having a record of their new equipment being validated. I am assume that the validation of the equipment means that they should have run a parallel test with the new and "old" piece of equipment to verify that it indeed performs as it should. The customer felt it was our responsibility to do the validation testing in their lab for the equipment. Does anyone have some more insight or protocols as to what equipment validation documentation is required of inspecting agencies like Joint Commission ? I appreciate your input. Have a Blessed Day ! Best regards, From sa.drew <@t> hosp.wisc.edu Wed Sep 1 12:20:26 2004 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Factor XIIIa Message-ID: We have been using BioCare Medical's polyclonal FXIIIa very successfully on the Ventana immunostainers. If you would be interested in more info on this feel free to contact me or BioCare... -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Wednesday, September 01, 2004 10:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Factor XIIIa Thanks for all the replys to my query about finding a new supplier for Factor XIIIa. I called DAKO and they told me that they DO NOT have Factor XIIIa. Someone yesterday emailed me and said they have a polyclonal Factor XIIIa that they think works very well. If youre out there, email me again with the product info please! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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 Xudong_Cao <@t> brown.edu Wed Sep 1 12:39:09 2004 From: Xudong_Cao <@t> brown.edu (Cao, Xudong) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] vector ABC Message-ID: <1DA88DDC48CCC245A777599FDAEED6D0A39092@MAIL1.AD.Brown.Edu> Dear all, does anybody know how long a ready-to-use ABC solution is good for? I am thinking about re-use the mixed solution if it is good for more than 48 hours... thanks. Xudong Cao post-doc fellow Brown University From ploykasek <@t> phenopath.com Wed Sep 1 13:21:32 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Equipment Validation In-Reply-To: Message-ID: I'm not familiar with the JCAO requirements, but I know a little about the GLP requirements. Usually these requirements are mainly intended for instruments that are reporting data. In anatomic path, the pathlogists are the "instrument". I do think it is smart to test any new equipment that comes into the lab to ensure it does what it is supposed to do. Patti Loykasek PhenoPath Laboratories Seattle, WA > Good morning everyone. > > I have a question concerning equipment validation. > > We recently had a customer that said they were cited during an inspection > for not having a record of their new equipment being validated. > I am assume that the validation of the equipment means that they should > have run a parallel test with the new and "old" piece of equipment to > verify that it indeed performs as it should. The customer felt it was our > responsibility to do the validation testing in their lab for the equipment. > Does anyone have some more insight or protocols as to what equipment > validation documentation is required of inspecting agencies like Joint > Commission ? > > I appreciate your input. > > Have a Blessed Day ! > > > Best regards, > > Mark > > Mark Lewis > Product Specialist > Anatomical Pathology > Clinical Diagnostics > Thermo Electron Corporation > (412) 747-4013 > (412) 788-1097 > E-mail: mark.lewis@thermo.com > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rpw4 <@t> psu.edu Wed Sep 1 13:24:34 2004 From: rpw4 <@t> psu.edu (Ronald P. Wilson) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Temperatures for IHC Message-ID: We are in the process of optimizing our IHC procedures. We have switched to using some of the reagents from Phoenix Biotechnologies. Their recommended protocols suggest primary and secondary antibody as well as strepavidin and chromagen incubations be done at 55?C for much shorter time periods (4-10 minutes). I have also heard that some labs do their incubations at 37?C. Is it common practice to use temperatures higher than room temperature? Are there advantages other than just shortening the incubation time? If anyone is using the higher temperature, how do you do this for bench-top procedures? I know that some of the staining stations such as Microprobe have built-in incubation chambers to do this. Ronald P. Wilson, V.M.D., M.S. Associate Professor Department of Comparative Medicine, HO54 Penn State University College of Medicine M. S. Hershey Medical Center 500 University Drive Hershey, PA 17033 phone: 717-531-8460 fax: 717-531-5001 e-mail: rpw4@psu.edu From Sandra.Etheridge <@t> gems8.gov.bc.ca Wed Sep 1 13:45:11 2004 From: Sandra.Etheridge <@t> gems8.gov.bc.ca (Etheridge, Sandra AGF:EX) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Alcohol/Xylene Recycling Message-ID: <424FAC08A8CDFE4BADD6497AE75B5E2F0E0A871D@atlas.gov.bc.ca> Hello, all, Does anyone have any info on alcohol and/or xylene recycling systems? We are currently having these two reagents treated off site and we are wondering if it is feasible to purchase a distiller, some sort of filter system, etc. What was your initial capital cost and your subsequent operating cost? Any problems with your current system? Any feedback, as always, is appreciated. Thanks in advance. Sandra Etheridge BC Ministry of Agriculture, Food & Fisheries Animal Health Center, Histology Abbotsford, BC Canada From mcauliff <@t> umdnj.edu Wed Sep 1 17:49:37 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] vector ABC In-Reply-To: <1DA88DDC48CCC245A777599FDAEED6D0A39092@MAIL1.AD.Brown.Edu> References: <1DA88DDC48CCC245A777599FDAEED6D0A39092@MAIL1.AD.Brown.Edu> Message-ID: <41365201.7070300@umdnj.edu> Why not call Vector and ask them? It is their product and they could give you the most accurate advice. For myself, I would mix fresh. Geoff Cao, Xudong wrote: >Dear all, > >does anybody know how long a ready-to-use ABC solution is good for? I am thinking about re-use the mixed solution if it is good for more than 48 hours... > >thanks. > >Xudong Cao >post-doc fellow >Brown University > >_______________________________________________ >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 gu.lang <@t> gmx.at Wed Sep 1 15:02:21 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] IF on FFPE renal biopsies Message-ID: <001a01c4905e$97a62470$eeeea8c0@server> Hi, after further looking for if-information I have seen, that performing immunofluorescence on paraffinslides is not really usual. Is there anybody in histoland, who has experience with this and can share a protocol? thanks in advance Gudrun Lang From AFeatherstone <@t> KaleidaHealth.Org Wed Sep 1 15:02:20 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] ZAP-70 Message-ID: Anyone using immunohistochemical methods with ZAP-70 antibody? What antibody are you using? Vendor , ETC? Thanks Annette Featherstone HT/MLT Kalieda Health -----Original Message----- From: Patti Loykasek [mailto:ploykasek@phenopath.com] Sent: Wednesday, September 01, 2004 14:22 To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Equipment Validation I'm not familiar with the JCAO requirements, but I know a little about the GLP requirements. Usually these requirements are mainly intended for instruments that are reporting data. In anatomic path, the pathlogists are the "instrument". I do think it is smart to test any new equipment that comes into the lab to ensure it does what it is supposed to do. Patti Loykasek PhenoPath Laboratories Seattle, WA > Good morning everyone. > > I have a question concerning equipment validation. > > We recently had a customer that said they were cited during an inspection > for not having a record of their new equipment being validated. > I am assume that the validation of the equipment means that they should > have run a parallel test with the new and "old" piece of equipment to > verify that it indeed performs as it should. The customer felt it was our > responsibility to do the validation testing in their lab for the equipment. > Does anyone have some more insight or protocols as to what equipment > validation documentation is required of inspecting agencies like Joint > Commission ? > > I appreciate your input. > > Have a Blessed Day ! > > > Best regards, > > Mark > > Mark Lewis > Product Specialist > Anatomical Pathology > Clinical Diagnostics > Thermo Electron Corporation > (412) 747-4013 > (412) 788-1097 > E-mail: mark.lewis@thermo.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 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 gu.lang <@t> gmx.at Wed Sep 1 15:40:58 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] IF on FFPE renal biopsies References: <001a01c4905e$97a62470$eeeea8c0@server> <6.0.0.22.1.20040901142551.01b33b78@gemini.msu.montana.edu> Message-ID: <002101c49063$fcd6bcb0$eeeea8c0@server> Gayle, I think performing IF on paraffin sections is not usual. Gudrun ----- Original Message ----- From: "Gayle Callis" To: "Gudrun Lang" Sent: Wednesday, September 01, 2004 10:28 PM Subject: Re: [Histonet] IF on FFPE renal biopsies > For what is not usual - do you mean frozen sections or for FFPE? Just > trying to sort out what you mean > > > > At 02:02 PM 9/1/2004, you wrote: > >Hi, > >after further looking for if-information I have seen, that performing > >immunofluorescence on paraffinslides is not really usual. > >Is there anybody in histoland, who has experience with this and can share > >a protocol? > >thanks in advance > > > >Gudrun Lang > >_______________________________________________ > >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 brett_connolly <@t> merck.com Wed Sep 1 15:51:09 2004 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Large section cryostat Message-ID: Is anyone using a Bright 8000 series or a Bright 9400 cryostat? I might be in the market for one in the near future and would like recommendations for these and/or any other manufacturer of large section cryos. Thx, 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 HornHV <@t> archildrens.org Wed Sep 1 16:02:37 2004 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] ZAP-70 Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322B93C@EMAIL.archildrens.org> I haven't used it but the Cell Marque rep came by today and they now have this antibody. If you want to call them their number is: 1-800-665-7284 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 Featherstone, Annette Sent: Wednesday, September 01, 2004 3:02 PM To: 'Patti Loykasek'; Histonet@lists.utsouthwestern.edu Subject: [Histonet] ZAP-70 Anyone using immunohistochemical methods with ZAP-70 antibody? What antibody are you using? Vendor , ETC? Thanks Annette Featherstone HT/MLT Kalieda Health -----Original Message----- From: Patti Loykasek [mailto:ploykasek@phenopath.com] Sent: Wednesday, September 01, 2004 14:22 To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Equipment Validation I'm not familiar with the JCAO requirements, but I know a little about the GLP requirements. Usually these requirements are mainly intended for instruments that are reporting data. In anatomic path, the pathlogists are the "instrument". I do think it is smart to test any new equipment that comes into the lab to ensure it does what it is supposed to do. Patti Loykasek PhenoPath Laboratories Seattle, WA > Good morning everyone. > > I have a question concerning equipment validation. > > We recently had a customer that said they were cited during an > inspection for not having a record of their new equipment being > validated. I am assume that the validation of the equipment means that > they should have run a parallel test with the new and "old" piece of > equipment to verify that it indeed performs as it should. The customer > felt it was our responsibility to do the validation testing in their > lab for the equipment. > Does anyone have some more insight or protocols as to what equipment > validation documentation is required of inspecting agencies like Joint > Commission ? > > I appreciate your input. > > Have a Blessed Day ! > > > Best regards, > > Mark > > Mark Lewis > Product Specialist > Anatomical Pathology > Clinical Diagnostics > Thermo Electron Corporation > (412) 747-4013 > (412) 788-1097 > E-mail: mark.lewis@thermo.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 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 ------------------------------------------------------------------------------ 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 JGordon <@t> cellmarque.com Wed Sep 1 16:31:00 2004 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Factor XIIIa Message-ID: Hi Angela, you may have gotten my e-mail. Factor 13a polyclonal seems to be in short supply, so we have had many labs turning to our monoclonal Factor 13a as a preferred alternative. The feedback that we are getting is that it has performed even better than the previous polyclonal antibody! I am including the data sheet and a photo of the stain for you to look over and share with your pathologist. Cell Marque's Factor 13a is 100% money back guaranteed to work. If you have any further questions, please contact me at 1-800-665-7284, Ext. 12. If you need any references that may not already frequent the Histonet (such as Johns Hopkins or LabCorp or Jackson Madison), please contact me and I can provide you with any information that you may need. Jeff Gordon Cell Marque Corp. 1-800-665-7284, Ext. 12 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Angela Bitting Sent: Wednesday, September 01, 2004 10:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Factor XIIIa Thanks for all the replys to my query about finding a new supplier for Factor XIIIa. I called DAKO and they told me that they DO NOT have Factor XIIIa. Someone yesterday emailed me and said they have a polyclonal Factor XIIIa that they think works very well. If youre out there, email me again with the product info please! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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 ihc <@t> unipathllc.com Wed Sep 1 16:55:45 2004 From: ihc <@t> unipathllc.com (UniPath IHC) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] NIST Certified Message-ID: <000001c4906e$6f19ca60$4500a8c0@unipathllc.corp> Hello Histoland, Does anybody know how to tell if your thermometric devices are "NIST certified" (CAP questions ANP23090, ANP23095)? Our thermometers (non-mercury, of course) don't mention this. Does this also apply to incubators/ovens, etc. with internal thermometers? Thanks, Brianna Jackson, BS QIHC UniPath, LLC Denver, CO From aostrand <@t> seattlecca.org Wed Sep 1 17:01:48 2004 From: aostrand <@t> seattlecca.org (Ostrander, Anita B) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Toxoplasma Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B94801D30ACE@wala01.seattlecca.org> I am looking for a vendor/company to purchase toxoplasma antibody from. Any suggestions? Thank you 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 joshua71 <@t> gmx.de Wed Sep 1 17:27:22 2004 From: joshua71 <@t> gmx.de (Joshua Tree) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] I have a funny References: Message-ID: <31472.1094077642@www26.gmx.net> Juan: YOU ARE A GREAT AMERICAN! > Been there, done that, and got my a.. chewed out by my boss when they > threatened legal action. Maybe if those companies spent less time on the > histonet and more time taking care of the customer this thread will go away. > NOW I DID NOT MENTION ANY NAMES SO DON'T CALL ME OR MY BOSS, OR YOUR LAWYERS, > OR MY LAWYERS, OR YOUR LAWYERS' LAWYER, OR YOUR COUSIN THE LAWYER. ALSO > DON'T HAVE YOUR PEOPLE CALL MY PEOPLE EITHER, I DON'T WANT TO DO LUNCH! Of > course I'm referring to the companies and not to you Lucy. > > Juan C. Gutierrez, HT(ASCP) > Histology Laboratory Supervisor > (210)704-2533 > > My views are my own and do not reflect those of my employer. > Long live free speech! "We have nothing to fear but fear itself". > > > -----Original Message----- > From: Lucy Brooks [mailto:lucyb@biocare.net] > Sent: Wednesday, September 01, 2004 10:23 AM > To: Sarah Jones; histonet@pathology.swmed.edu; JNocito@Pathreflab.com > Subject: Re: [Histonet] I have a funny > > > I personally hope the histonet isn't used to "intimidate companies" , but > I > do feel that if you have a problem and the company who you have that > problem > with doesn't deal with it in a reasonable amount of time, it is nice to > warn > other people out their of what they can expect when dealing with these > said > companies. > ----- Original Message ----- > From: "Sarah Jones" > To: ; ; > > Sent: Tuesday, August 31, 2004 6:56 PM > Subject: Re: [Histonet] I have a funny > > > > Ah, this begs an ethical question.... should the histonet be used as a > > threat to intimidate companies? > > Is this an Us against Them mentality? > > Discuss amongst yourselves. (a little "Coffee Talk" humor there for > > you Saturday Night Live fans) > > Oye Vey..................... Sarah > > > > > > Sarah Jones, HT(ASCP) > > Histology Lab > > Dept. of Veterinary Integrative Biosciences > > College of Veterinary Medicine > > Texas A&M University > > College Station, TX 77843-4458 > > phone: 979-845-3177 > > fax: 979-458-3499 > > > > > > >>> "Lucy Brooks" 8/31/2004 6:10:52 PM >>> > > Joe!! > > > > Speak your mind!!! > > The Histonet is a powerful thing!! > > > > ----- Original Message ----- > > From: "Joe Nocito" > > To: "Histonet" > > Sent: Tuesday, August 31, 2004 4:01 PM > > Subject: [Histonet] I have a funny > > > > > > > Hello all, > > > for once, I have, what I think is funny, is a short story. A couple > > of > > > months ago, I purchased a piece of equipment from a vendor (who will > > remain > > > anonymous, at this time at least). I was told it was 6 weeks for > > delivery. > > > It's been over 8 weeks. I called the rep and left a voice mail asking > > when > > I > > > can expect my equipment. S/he called me right back. I asked when I > > can > > > expect the equipment. Then I said "Don't make me go on the > > Histonet". > > > My techs started laughing and I couldn't keep a straight face. The > > rep was > > > all apologetic and promised me I'll have my equipment in 2 weeks. > > Gee, I > > > didn't think I was getting that bad. Maybe I should settle down a > > little > > > bit. > > > > > > 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 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- NEU: Bis zu 10 GB Speicher für e-mails & Dateien! 1 GB bereits bei GMX FreeMail http://www.gmx.net/de/go/mail From Rcartun <@t> harthosp.org Wed Sep 1 18:05:53 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Toxoplasma Message-ID: BioGenex Laboratories San Ramon, CA 94583 (800) 421-4149 RWC Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Ostrander, Anita B" 09/01/04 06:01PM >>> I am looking for a vendor/company to purchase toxoplasma antibody from. Any suggestions? Thank you 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rd <@t> medlabsouth.co.nz Wed Sep 1 21:06:43 2004 From: rd <@t> medlabsouth.co.nz (Roger Davies) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Re: Overuse injury from Microtomy Message-ID: <000e01c49091$979fb3a0$168524ca@rd> Hi All, I am writing to ask if anyone can provide me with information on RSI, OOS or Work related Musculo-skeletal injury associated with the microtome. I think we in histopathology all know of someone, or have personal knowledge of such injury, but perhaps we work on and say nothing! I have some staff members who suffer badly form epicondylitis and carpel tunnel syndrome which is obviously related to microtomy, however, our national ACC will not accept it as a work related injury as they say there is no official documented evidence to support it. Does anyone know of websites or organisations, countries where this is accepted as a work related condition where I obtain "official" documentation !! Thanks in anticipation Roger From Gervaip <@t> aol.com Wed Sep 1 21:59:47 2004 From: Gervaip <@t> aol.com (Gervaip@aol.com) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Re: Overuse injury from Microtomy Message-ID: <1c1.1dacdac2.2e67e6a3@aol.com> Rodger, try site http://www.niehs.nih.gov/odhsb/ergoguid/chapii.htm And you are correct, many of us in the field suffer from repetitive motion disorders. To get the laboratory managers to acknowledge this and release monies for the proper equipment, chairs and workspaces is another thing. Good luck to you. Pearl Gervais New Orleans From Gervaip <@t> aol.com Wed Sep 1 22:05:44 2004 From: Gervaip <@t> aol.com (Gervaip@aol.com) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Re: Overuse injury from Microtomy Message-ID: <104.4fbcdcba.2e67e808@aol.com> Roger, I failed to mention that many of these problems can be avoided with the use of proper posture and movements (ergonomics), adjustments to the work area and microtome , work arrangement and use of a good chair. I have some information you might find useful. Let me know if you want me to mail it to you. Pearl Gervais From beyer_janina <@t> yahoo.com.au Thu Sep 2 00:44:25 2004 From: beyer_janina <@t> yahoo.com.au (=?iso-8859-1?q?Janina=20Beyer?=) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Histology Resin Message-ID: <20040902054425.2096.qmail@web14526.mail.yahoo.com> Hi I am a newcomer and have just discovered Histonet. I have started an honours thesis in Tasmania, Australia, and will be looking at sex differentiation in the common carp. I have been told that paraffin is not the best embedding method for fish embryos and problems with sectioning the yolk part occurs. It was recommended to me to use methyl methacrylate monomer and Benzoyl peroxide as a medium. I was wondering if anyone had used this for embedding embryos, and weather it is recommended? I am also looking at undertaking some immunohistochemistry with these samples, will that be a problem? I would be grateful for any advice, or even references that you can suggest that may be helpful. Best wishes, Janina Janina Beyer Honours Student University of Tasmania CSIRO Marine Research Hobart, Tasmania Scientists were rated as great heretics by the church, but they were truly religious men because of their faith in the orderliness of the universe. --Albert Einstein --------------------------------- Find local movie times and trailers on Yahoo! Movies. From Sarah.Blachford <@t> ngh.nhs.uk Thu Sep 2 04:27:33 2004 From: Sarah.Blachford <@t> ngh.nhs.uk (Blachford, Sarah - Histopath Main) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Warthin Starey Message-ID: <854FA06D8BE2D611ABA30030052D1DF6FA58F2@NGH_EXCHANGE1> Can anyone help us. Has anyone got a reliable warthin starey method. We have tried many methods but cannot get them to work. Also what is the use of gelatin in the method Sarah Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD This e-mail may contain confidential information and/or copyright material and is intended for the use of the addressee only. Any unauthorised use may be unlawful. The contents of this e-mail may be subject to public disclosure under the NHS Code of Openness or the Freedom of Information Act 2000. Unless legally exempt, the confidentiality of the message and your reply cannot be guaranteed. If you receive this e-mail by mistake, please advise the sender immediately. Thank you. From Krat18 <@t> aol.com Thu Sep 2 05:31:19 2004 From: Krat18 <@t> aol.com (Krat18@aol.com) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Warthin Starey Message-ID: <55.60836828.2e685077@aol.com> Our lab has gone to the Steiner method because we get more consistent results. Karen Raterman From lpwenk <@t> sbcglobal.net Thu Sep 2 06:34:06 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Re: Overuse injury from Microtomy References: <000e01c49091$979fb3a0$168524ca@rd> Message-ID: <001501c490e0$c283ad20$3039d445@domainnotset.invalid> Lab Ergonomic sites: http://www.prevencionintegral.com/Articulos/@Datos/02_104.htm Australia study of histotechs and RMS http://www.niehs.nih.gov/odhsb/ergoguid/home.htm Lab ergonomics - National Institute of Health (NIH) http://www.cdc.gov/od/ohs/Ergonomics/labergo.htm Lab ergonomics - Center for Disease Control (CDC) Good luck. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Roger Davies" To: Sent: Wednesday, September 01, 2004 10:06 PM Subject: [Histonet] Re: Overuse injury from Microtomy Hi All, I am writing to ask if anyone can provide me with information on RSI, OOS or Work related Musculo-skeletal injury associated with the microtome. I think we in histopathology all know of someone, or have personal knowledge of such injury, but perhaps we work on and say nothing! I have some staff members who suffer badly form epicondylitis and carpel tunnel syndrome which is obviously related to microtomy, however, our national ACC will not accept it as a work related injury as they say there is no official documented evidence to support it. Does anyone know of websites or organisations, countries where this is accepted as a work related condition where I obtain "official" documentation !! Thanks in anticipation Roger _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From failm <@t> musc.edu Thu Sep 2 06:47:43 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Warthin Starey Message-ID: Sarah, We use the following method with good results though the Steiner is used more often some of our pathologists prefer the Warthin-Starry. Solutions needed 1% Silver Nitrate 2% Silver Nitrate 0.15% Hydroquinone 5% Gelatin (use gentle heat to mix) All of the above are stored at 2-8 degrees Celsius for up to 6 months. !. Preheat a coplin jar with 1% silver nitrate in a 54-60 degree water bath for 30 minutes. Place your gelatin bottle in the water bath at the same time. 2. Deparaffinize and hydrate slides to distilled water 3. Place slides in the preheated 1% Silver Nitrate and incubate for 30 minutes in a 54-60 degree water bath. At the same time place a vial of 1.5 mls of 2% silver nitrate, a vial of 3.75 5% gelatin, and a vial of 2 mls of 0.15% hydroquinone into the 54-60 degree waterbath to preheat.for 30 minutes. 4. Prepare the developer solution by mixing the solutions in the following order. 1.5 mls of 2% silver Nitrate 3.75mls of 5 % gelatin and 2.0 mls of 0.15% hydroquinone 5. Remove slides from heated silver (DO NOT RINSE) Place slides horizontally on a paper towel and cover with developer. Allow sections to develop until they are a light golden brown. Check microscopically for end point 6. Rinse thoroughly in hot tap water 7. Dehydrate, clear and mount Rena Fail Medical University of SC >>> "Blachford, Sarah - Histopath Main" 09/02/04 05:27AM >>> Can anyone help us. Has anyone got a reliable warthin starey method. We have tried many methods but cannot get them to work. Also what is the use of gelatin in the method Sarah Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD This e-mail may contain confidential information and/or copyright material and is intended for the use of the addressee only. Any unauthorised use may be unlawful. The contents of this e-mail may be subject to public disclosure under the NHS Code of Openness or the Freedom of Information Act 2000. Unless legally exempt, the confidentiality of the message and your reply cannot be guaranteed. If you receive this e-mail by mistake, please advise the sender immediately. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Marie-Helene.Lavergne <@t> mdsps.com Thu Sep 2 08:22:00 2004 From: Marie-Helene.Lavergne <@t> mdsps.com (Marie-Helene.Lavergne@mdsps.com) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Blade holder and disposable blade for Cryostat Message-ID: I'm looking for blade holder with disposable blades that can fit on a Bright 8000 cryostat. Does someone know where I can find these? Thank you Marie-Helene Lavergne DMPK MDSPS Canada From funderwood <@t> mcohio.org Thu Sep 2 08:38:51 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:23:59 2005 Subject: [BULK] - Re: [Histonet] I have a funny Message-ID: I wonder if the threat of going to histonet will work help in disagreements with my wife? >>> "Sarah Jones" 08/31/04 09:56PM >>> Ah, this begs an ethical question.... should the histonet be used as a threat to intimidate companies? Is this an Us against Them mentality? Discuss amongst yourselves. (a little "Coffee Talk" humor there for you Saturday Night Live fans) Oye Vey..................... Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Lucy Brooks" 8/31/2004 6:10:52 PM >>> Joe!! Speak your mind!!! The Histonet is a powerful thing!! ----- Original Message ----- From: "Joe Nocito" To: "Histonet" Sent: Tuesday, August 31, 2004 4:01 PM Subject: [Histonet] I have a funny > Hello all, > for once, I have, what I think is funny, is a short story. A couple of > months ago, I purchased a piece of equipment from a vendor (who will remain > anonymous, at this time at least). I was told it was 6 weeks for delivery. > It's been over 8 weeks. I called the rep and left a voice mail asking when I > can expect my equipment. S/he called me right back. I asked when I can > expect the equipment. Then I said "Don't make me go on the Histonet". > My techs started laughing and I couldn't keep a straight face. The rep was > all apologetic and promised me I'll have my equipment in 2 weeks. Gee, I > didn't think I was getting that bad. Maybe I should settle down a little > bit. > > 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Thu Sep 2 08:48:54 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:59 2005 Subject: [BULK] - Re: [Histonet] I have a funny Message-ID: Disagreements with your spouse? What are those? My husband knows I'm always right - I'm the one who is an expert with knives. (ASCP) "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/02/2004 08:38 AM To: cc: Subject: [BULK] - Re: [Histonet] I have a funny I wonder if the threat of going to histonet will work help in disagreements with my wife? >>> "Sarah Jones" 08/31/04 09:56PM >>> Ah, this begs an ethical question.... should the histonet be used as a threat to intimidate companies? Is this an Us against Them mentality? Discuss amongst yourselves. (a little "Coffee Talk" humor there for you Saturday Night Live fans) Oye Vey..................... Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Lucy Brooks" 8/31/2004 6:10:52 PM >>> Joe!! Speak your mind!!! The Histonet is a powerful thing!! ----- Original Message ----- From: "Joe Nocito" To: "Histonet" Sent: Tuesday, August 31, 2004 4:01 PM Subject: [Histonet] I have a funny > Hello all, > for once, I have, what I think is funny, is a short story. A couple of > months ago, I purchased a piece of equipment from a vendor (who will remain > anonymous, at this time at least). I was told it was 6 weeks for delivery. > It's been over 8 weeks. I called the rep and left a voice mail asking when I > can expect my equipment. S/he called me right back. I asked when I can > expect the equipment. Then I said "Don't make me go on the Histonet". > My techs started laughing and I couldn't keep a straight face. The rep was > all apologetic and promised me I'll have my equipment in 2 weeks. Gee, I > didn't think I was getting that bad. Maybe I should settle down a little > bit. > > 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 _______________________________________________ Histonet mailing list Histonet@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 Thu Sep 2 09:03:46 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:23:59 2005 Subject: [BULK] - Re: [Histonet] I have a funny Message-ID: And my knows that I have access to red barrels, and that I'm in charge of the morgue. :) -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: Thursday, September 02, 2004 8:49 AM To: Fred Underwood Cc: histonet@pathology.swmed.edu Subject: Re: [BULK] - Re: [Histonet] I have a funny Disagreements with your spouse? What are those? My husband knows I'm always right - I'm the one who is an expert with knives. (ASCP) "Fred Underwood" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/02/2004 08:38 AM To: cc: Subject: [BULK] - Re: [Histonet] I have a funny I wonder if the threat of going to histonet will work help in disagreements with my wife? >>> "Sarah Jones" 08/31/04 09:56PM >>> Ah, this begs an ethical question.... should the histonet be used as a threat to intimidate companies? Is this an Us against Them mentality? Discuss amongst yourselves. (a little "Coffee Talk" humor there for you Saturday Night Live fans) Oye Vey..................... Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Lucy Brooks" 8/31/2004 6:10:52 PM >>> Joe!! Speak your mind!!! The Histonet is a powerful thing!! ----- Original Message ----- From: "Joe Nocito" To: "Histonet" Sent: Tuesday, August 31, 2004 4:01 PM Subject: [Histonet] I have a funny > Hello all, > for once, I have, what I think is funny, is a short story. A couple of > months ago, I purchased a piece of equipment from a vendor (who will remain > anonymous, at this time at least). I was told it was 6 weeks for delivery. > It's been over 8 weeks. I called the rep and left a voice mail asking when I > can expect my equipment. S/he called me right back. I asked when I can > expect the equipment. Then I said "Don't make me go on the Histonet". > My techs started laughing and I couldn't keep a straight face. The rep was > all apologetic and promised me I'll have my equipment in 2 weeks. Gee, I > didn't think I was getting that bad. Maybe I should settle down a little > bit. > > 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 _______________________________________________ Histonet mailing list Histonet@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 dennijc <@t> vetmed.auburn.edu Thu Sep 2 09:54:04 2004 From: dennijc <@t> vetmed.auburn.edu (John C. Dennis) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] vector ABC In-Reply-To: <1DA88DDC48CCC245A777599FDAEED6D0A39092@MAIL1.AD.Brown.Edu> References: <1DA88DDC48CCC245A777599FDAEED6D0A39092@MAIL1.AD.Brown.Edu> Message-ID: The Vector tech folk once told me that they'd used it up to several hours (8-12). I've kept it up to 4 hours but after that I dump it. John Carroll Dennis Anatomy, Physiology, and Pharmacology 109 Greene Hall Auburn University, AL 36849 On Wed, 1 Sep 2004, Cao, Xudong wrote: > Dear all, > > does anybody know how long a ready-to-use ABC solution is good for? I am thinking about re-use the mixed solution if it is good for more than 48 hours... > > thanks. > > Xudong Cao > post-doc fellow > Brown University > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ritta69 <@t> iwon.com Thu Sep 2 10:03:30 2004 From: ritta69 <@t> iwon.com (ritta69@iwon.com) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Timm stain Message-ID: <20040902150330.9F79F124B8@email.iwon.com> Has anyone out there done a timm stain for mossy fiber sprouting? _______________________________________________ From pmarcum <@t> polysciences.com Thu Sep 2 10:39:40 2004 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Timm stain In-Reply-To: <20040902150330.9F79F124B8@email.iwon.com> Message-ID: <001101c49103$0fad94e0$4f00a8c0@PMARCUM2K> Yes Years ago and I still remember it painfully!! Thanks, Pam Marcum? > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of > ritta69@iwon.com > Sent: Thursday, September 02, 2004 11:04 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Timm stain > > > > > > > > > > Has anyone out there done a timm stain for mossy fiber sprouting? > > _______________________________________________ > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From dmccaig <@t> ckha.on.ca Thu Sep 2 10:58:39 2004 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] sSILVER STAIN FOR CAT SCRATCH FEVER Message-ID: <3E5A3F039F0BD8118B4700C00D002024043282@CKHA9> Hi I was wondering if anyone could suggest what is the recommended special stain to demonstrate Cat Scratch Fever (coccibacilli) in a lymph node? Diana McCaig, R.T. From gcallis <@t> montana.edu Thu Sep 2 11:05:13 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] TImm stain Message-ID: <6.0.0.22.1.20040902100429.01afa818@gemini.msu.montana.edu> Go to Histonet Archives, this protocol has been posted before. www.histosearch.org 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 JQB7 <@t> CDC.GOV Thu Sep 2 11:07:06 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] sSILVER STAIN FOR CAT SCRATCH FEVER Message-ID: We use the Modified Steiner with slightly adjusted times for this organism. We also use IHC. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Diana McCaig Sent: Thursday, September 02, 2004 11:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] sSILVER STAIN FOR CAT SCRATCH FEVER Hi I was wondering if anyone could suggest what is the recommended special stain to demonstrate Cat Scratch Fever (coccibacilli) in a lymph node? Diana McCaig, R.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From failm <@t> musc.edu Thu Sep 2 11:10:14 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] sSILVER STAIN FOR CAT SCRATCH FEVER Message-ID: Warthin-Starry or Steiner Rena Fail >>> Diana McCaig 09/02/04 11:58AM >>> Hi I was wondering if anyone could suggest what is the recommended special stain to demonstrate Cat Scratch Fever (coccibacilli) in a lymph node? Diana McCaig, R.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcokie <@t> yahoo.com Thu Sep 2 11:13:44 2004 From: jcokie <@t> yahoo.com (Jeanenne Duffy) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Potomac Horse Fever antibody? Message-ID: <20040902161344.97660.qmail@web53510.mail.yahoo.com> Dear Histonet: Is anyone running IHC for Potomac Horse Fever? I am looking for an antibody to use on FFPE tissues. If there are any sources out there, please post. Thank-you very much! Jeanenne Duffy Oklahoma Animal Disease Diagnostic Lab Stillwater, OK ph 405-744-6623 _______________________________ Do you Yahoo!? Win 1 of 4,000 free domain names from Yahoo! Enter now. http://promotions.yahoo.com/goldrush From sharon.willman <@t> bms.com Thu Sep 2 11:15:53 2004 From: sharon.willman <@t> bms.com (Sharon E Willman) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] BrdU Injections Message-ID: <41374739.3050500@bms.com> Hi, I was wondering if anyone has had issues with rats injected with BrdU and 1 hour later were necropsied. Should the incubation time of Brdu be extended. The positive control on the stainer does have results of being positive. We use duodenum as an internal control from each rat. We are seeing a small percentage of rats that are not staining positive on the internal control. Anyone with any suggestions would be most appreciated. Thanks in advance! Sharon Willman From shive003 <@t> umn.edu Thu Sep 2 11:44:06 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] IHC for Canine Distemper Virus Message-ID: <005801c4910c$100589d0$78065486@vdl220FAC> I would greatly appreciate any vendor source suggestions for Canine Distemper Virus antibody (for use in IHC on FFPE tissue). Thank you very much in advance. Jan Shivers IHC Supervisor Univ. of Minnesota Veterinary Diagnostic Lab 1333 Gortner Ave. St. Paul, MN 55108 shive003@umn.edu From David.Edmondson <@t> christie-tr.nwest.nhs.uk Thu Sep 2 11:47:56 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] vector ABC to Cao Xudong Message-ID: To Cao Xudong/co Histonet The DAKO ABC kit we used after several days, I thought they suggested a week by way of a use-by time. But that is a few years back now. Dave Christie Manchester UK -----Original Message----- From: John C. Dennis [mailto:dennijc@vetmed.auburn.edu] Sent: 02 September 2004 15:54 To: Cao, Xudong Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] vector ABC The Vector tech folk once told me that they'd used it up to several hours (8-12). I've kept it up to 4 hours but after that I dump it. John Carroll Dennis Anatomy, Physiology, and Pharmacology 109 Greene Hall Auburn University, AL 36849 On Wed, 1 Sep 2004, Cao, Xudong wrote: > Dear all, > > does anybody know how long a ready-to-use ABC solution is good for? I am thinking about re-use the mixed solution if it is good for more than 48 hours... > > thanks. > > Xudong Cao > post-doc fellow > Brown 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 jtsonger <@t> vt.edu Thu Sep 2 12:42:12 2004 From: jtsonger <@t> vt.edu (Jill Songer) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Potomac Horse Fever antibody? In-Reply-To: <20040902161344.97660.qmail@web53510.mail.yahoo.com> References: <20040902161344.97660.qmail@web53510.mail.yahoo.com> Message-ID: <6.0.0.22.0.20040902134007.01b1e100@pop.vt.edu> If you find out if any body does this, please let me know. We had a case a few weeks ago and I tried to find someone who does this, as we don't run IHC. Ended up doing a Modified Steiner which turned out OK, but I think the pathologists would have preferred IHC. Thanks At 12:13 PM 9/2/2004, Jeanenne Duffy wrote: >Dear Histonet: >Is anyone running IHC for Potomac Horse Fever? I am >looking for an antibody to use on FFPE tissues. If >there are any sources out there, please post. >Thank-you very much! > >Jeanenne Duffy >Oklahoma Animal Disease Diagnostic Lab >Stillwater, OK > >ph 405-744-6623 > > > >_______________________________ >Do you Yahoo!? >Win 1 of 4,000 free domain names from Yahoo! Enter now. >http://promotions.yahoo.com/goldrush > >_______________________________________________ >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 kosmicdog <@t> hotmail.com Thu Sep 2 13:34:40 2004 From: kosmicdog <@t> hotmail.com (jason m) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] blade sign Message-ID: anyone have tips for minimizing blade sign? i start at one end of fresh blade and work to the other end before throwing out the disposable blade. sometimes however, blade sign appears even with a new blade. is it just that some blocks have hard spots that nick the blade? thanks. J. _________________________________________________________________ Take advantage of powerful junk e-mail filters built on patented Microsoft® SmartScreen Technology. http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines Start enjoying all the benefits of MSN® Premium right now and get the first two months FREE*. From Jackie.O'Connor <@t> abbott.com Thu Sep 2 13:39:53 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Cerebral bleeds in mice - please hurry! Message-ID: I've been asked to section mice brains to look for possible cerebral bleeds - They're harvesting the brains intact - can anyone suggest the best way to bisect or trisect them after fixation to look for small hemorrhages? I usually trisect transversely through the cerebellum, cerebral hemisphere, and olfactory bulb for routine looks at the brain, but is there a better method for looking for bleeds? They should see evidence at necropsy, I hope - but if not, it's all me me me. Suggestions PLEASE. Jackie O' P.S. Good wishes to all you Floridians who may be impacted by Frances in the next 48 hours. I hope your're not at work anymore. From jcokie <@t> yahoo.com Thu Sep 2 13:56:00 2004 From: jcokie <@t> yahoo.com (Jeanenne Duffy) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Canine Distemper antibody source Message-ID: <20040902185600.49171.qmail@web53510.mail.yahoo.com> Our lab uses a monoclonal for Canine Distemper Virus from Custom Monoclonals International. Their ph # is 916-375-1139. The catalog # is DV2-12. It works very well with HIER at a 1:2000 dilution. Jeanenne Duffy Oklahoma Animal Disease Diagnostic Lab Stillwater, OK ph 405-744-6623 _______________________________ Do you Yahoo!? Win 1 of 4,000 free domain names from Yahoo! Enter now. http://promotions.yahoo.com/goldrush From gcallis <@t> montana.edu Thu Sep 2 15:06:14 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Cerebral bleeds in mice - please hurry! In-Reply-To: References: Message-ID: <6.0.0.22.1.20040902140201.01b296d8@gemini.msu.montana.edu> Buy a brain matrix so you can do precise slides of the whole brain. These are available at www.Myneurolab.com. Very tidy - we use high profile microtome blades - the ones we do not like! or a teflon coated razor blade from Ted Pella gives smooth, clean slicing. These devices allow you to bisect either direction and at different thicknesses. You may be able to do the slices (since the brain is held in place) while the brain is fresh as long as you Slice and do not PUSH down on top of brain. Good luck At 12:39 PM 9/2/2004, you wrote: >I've been asked to section mice brains to look for possible cerebral >bleeds - >They're harvesting the brains intact - can anyone suggest the best way to >bisect or trisect them after fixation to look for small hemorrhages? > >I usually trisect transversely through the cerebellum, cerebral >hemisphere, and olfactory bulb for routine looks at the brain, but is >there a better method for looking for bleeds? They should see evidence at >necropsy, I hope - but if not, it's all me me me. > >Suggestions PLEASE. > >Jackie O' > >P.S. Good wishes to all you Floridians who may be impacted by Frances in >the next 48 hours. I hope your're not at work anymore. >_______________________________________________ >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 Thu Sep 2 16:36:26 2004 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] muscle actin Message-ID: Has anyone used Dako's Muscle Actin HHF35 Code no. M0635 on mouse tissue? I realize that I will have to use m on m detection, I am just curious about this working on mouse or not. If you have this optimized please share your procedure with me including pretreatment if any for ffpe mouse lungs. Thank you, Patsy Ruegg From pruegg <@t> ihctech.net Thu Sep 2 16:36:29 2004 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] alkaline phosphatase staining In-Reply-To: <1E0CC447E59C974CA5C7160D2A2854EC03C40E75@SJMEMXMB04.stjude.sjcrh.local> Message-ID: the enzyme alk.phos. stain for osteoblasts will not work on decalcified ffpe tissues in my experience. it will work limitedly on bone processed into GMA without decal and cold methanol fixed. I hear that there are antibodies to alk.phos. but I have not used them yet, has anyone else? Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Garlits, John Sent: Monday, August 30, 2004 11:04 AM To: HistoNet Server Subject: [Histonet] alkaline phosphatase staining Hello, I am interested in doing some alkaline phosphatase staining in decalcified, paraffin-fixed mouse long bone sections. Does anyone have any experience with alk phos staining, who could recommend a protocol or even a commercial kit? Thank you! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 Phone: 901-495-3961 Fax: 901-495-2176 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Sep 2 16:36:55 2004 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] injecting tracers In-Reply-To: <6.1.0.6.2.20040830110655.024a0a08@tigger.uic.edu> Message-ID: Try a comany called Molecular Probes. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Lu Leach Sent: Monday, August 30, 2004 9:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] injecting tracers Dear Histonetters, I need advice on a fluorescent tracer to use for mouse injections that will hopefully survive tissue processing. If anyone has experience and wisdom in this area, please contact me. Thank you, Lu Leach University of Illinois at Chicago lleach@uic.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From northma <@t> ohsu.edu Thu Sep 2 17:03:53 2004 From: northma <@t> ohsu.edu (Mary North) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Modified muscle stain Message-ID: There is a modification of the succinic dehydrogenase stain that adds phenazine methosulfate in order to demonstrate abnormal mitochondria. I would appreciate receiving the technique if any of you have access to it. Mary North, HT(ASCP),HTL Neuromuscular Laboratory Oregon Health & Science University Portland, OR From David.Edmondson <@t> christie-tr.nwest.nhs.uk Fri Sep 3 04:39:01 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] RE: DAKO or VECTOR Message-ID: K0377, is a kit where a drop of HRP and a drop of Streptavidin get mixed together in buffer, The data sheet that I have from 1997 makes no mention of lifespan, now that I look Dave -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 02 September 2004 20:57 To: Edmondson David (RBV) NHS Christie Tr Subject: DAKO or VECTOR Does DAKO have an avidin/bioin complex kit, I thought they has Labelled Strepavidin aka LSAB? The ABC kit I am aware of is VECTOR's. At 10:47 AM 9/2/2004, you wrote: >To Cao Xudong/co Histonet >The DAKO ABC kit we used after several days, I thought they suggested a week >by way of a use-by time. But that is a few years back now. > >Dave >Christie >Manchester UK >-----Original Message----- >From: John C. Dennis [mailto:dennijc@vetmed.auburn.edu] >Sent: 02 September 2004 15:54 >To: Cao, Xudong >Cc: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] vector ABC > > > >The Vector tech folk once told me that they'd used it up to several hours >(8-12). I've kept it up to 4 hours but after that I dump it. > >John Carroll Dennis >Anatomy, Physiology, and Pharmacology >109 Greene Hall >Auburn University, AL 36849 > > >On Wed, 1 Sep 2004, Cao, Xudong wrote: > > > Dear all, > > > > does anybody know how long a ready-to-use ABC solution is good for? I am >thinking about re-use the mixed solution if it is good for more than 48 >hours... > > > > thanks. > > > > Xudong Cao > > post-doc fellow > > Brown 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 > >_______________________________________________ >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 flemons <@t> bhset.org Fri Sep 3 09:15:13 2004 From: flemons <@t> bhset.org (Fran Lemons) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Pre-cut controls Message-ID: Does anyone know why IHC control slides cannot be cut ahead of time? I've done it at other facilities, but I have been told it is not permissible here, without any explanation. Any ideas? Also, has anyone ever heard of refrigerating pre-cut IHC control slides? One person suggested to me that they stopped cutting them ahead of time because they were getting condensation on them & the stains wouldn't work. Then she informed me that they were stored in the fridge. Seems to me the condensation would have formed upon removing the slides from a cold environment to a room temp. Your thoughts/theories are appreciated. Fran Walker Histology Technical Specialist ETBH Knoxville From DonnaWillis <@t> texashealth.org Fri Sep 3 09:38:42 2004 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Pre-cut controls Message-ID: We precut our controls and place them in the refrigerator. We do not dry them until they are taken out for use. The only issue we have found is with Her-2-neu and what we do is only cut enough for 2 weeks use. Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fran Lemons Sent: Friday, September 03, 2004 9:15 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Pre-cut controls Does anyone know why IHC control slides cannot be cut ahead of time? I've done it at other facilities, but I have been told it is not permissible here, without any explanation. Any ideas? Also, has anyone ever heard of refrigerating pre-cut IHC control slides? One person suggested to me that they stopped cutting them ahead of time because they were getting condensation on them & the stains wouldn't work. Then she informed me that they were stored in the fridge. Seems to me the condensation would have formed upon removing the slides from a cold environment to a room temp. Your thoughts/theories are appreciated. Fran Walker Histology Technical Specialist ETBH Knoxville _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From MAUGER <@t> email.chop.edu Fri Sep 3 09:40:29 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Pre-cut controls Message-ID: Fran, Some antigenicity can be lost when cut slides are stored for a long time. Time varies for different antigens, and some are not effected at all. Some people think refrigeration, or even freezing controls slides lengthens storage time. I personally think that it's best to cut what you will use in a few months time. Trial and error. Jo >>> "Fran Lemons" 09/03/04 10:15AM >>> Does anyone know why IHC control slides cannot be cut ahead of time? I've done it at other facilities, but I have been told it is not permissible here, without any explanation. Any ideas? Also, has anyone ever heard of refrigerating pre-cut IHC control slides? One person suggested to me that they stopped cutting them ahead of time because they were getting condensation on them & the stains wouldn't work. Then she informed me that they were stored in the fridge. Seems to me the condensation would have formed upon removing the slides from a cold environment to a room temp. Your thoughts/theories are appreciated. Fran Walker Histology Technical Specialist ETBH Knoxville _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Fri Sep 3 09:44:38 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Pre-cut controls Message-ID: Fran, Some epitopes tend to lose their antigenicity due to oxidation in cut paraffin sections. We precut all our control slides and some, i.e. ER, PR, Ki67, etc. are stored in a -20 degree freezer until needed. Others, that seem to be stable, are stored at R.T. Once we figured out which antibody controls were stable and which weren't and stored them accordingly, we had no issues regarding loss of staining intensity. 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: Fran Lemons [mailto:flemons@bhset.org] Sent: Friday, September 03, 2004 9:15 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Pre-cut controls Does anyone know why IHC control slides cannot be cut ahead of time? I've done it at other facilities, but I have been told it is not permissible here, without any explanation. Any ideas? Also, has anyone ever heard of refrigerating pre-cut IHC control slides? One person suggested to me that they stopped cutting them ahead of time because they were getting condensation on them & the stains wouldn't work. Then she informed me that they were stored in the fridge. Seems to me the condensation would have formed upon removing the slides from a cold environment to a room temp. Your thoughts/theories are appreciated. Fran Walker Histology Technical Specialist ETBH Knoxville _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akbitting <@t> geisinger.edu Fri Sep 3 09:46:01 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Paraffin all over my floor....... Message-ID: We use a wax scraper with a long handle from MarketLab to clean up the paraffin stuck to the floor. It comes with replaceable blades. BUT someone said they saw a wider scraper somewhere in a catalog and we can't remember where we saw it. Does anyone out there have any ideas. The scraper we're looking for is about 6 inches wide. Thanx Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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 akbitting <@t> geisinger.edu Fri Sep 3 09:48:47 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Magnification Message-ID: Our Derm Path doc wants me to poll the Histonet subscribers about what everyone uses when they're embedding tiny tissues such as derms. Any creative magnifiers out there? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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 Bauer.Karen <@t> mayo.edu Fri Sep 3 09:55:03 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Pre-cut controls Message-ID: <8C6E05FA69571948B461F1327CBB893E0629F127@lmmail2.ad.lmmhs.org> We cut all of our controls ahead of time and have had no problems. Controls are tested before we use them and then of course are monitored when we actually use them. As far as I know, the Pathologists are happy with what we have and I have not had to change our way of handling controls. After we cut the controls, we put them in slide boxes and leave them open to air dry for a few hours or even overnight if cut late in the afternoon. We then place them in the cupboard and then use them when we need them. Good Luck to you... Karen Bauer HT(ASCP) Histology Supervisor Department of Pathology Luther Hospital Eau Claire, WI -----Original Message----- From: Joanne Mauger [mailto:MAUGER@email.chop.edu] Sent: Friday, September 03, 2004 9:40 AM To: flemons@bhset.org; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Pre-cut controls Fran, Some antigenicity can be lost when cut slides are stored for a long time. Time varies for different antigens, and some are not effected at all. Some people think refrigeration, or even freezing controls slides lengthens storage time. I personally think that it's best to cut what you will use in a few months time. Trial and error. Jo >>> "Fran Lemons" 09/03/04 10:15AM >>> Does anyone know why IHC control slides cannot be cut ahead of time? I've done it at other facilities, but I have been told it is not permissible here, without any explanation. Any ideas? Also, has anyone ever heard of refrigerating pre-cut IHC control slides? One person suggested to me that they stopped cutting them ahead of time because they were getting condensation on them & the stains wouldn't work. Then she informed me that they were stored in the fridge. Seems to me the condensation would have formed upon removing the slides from a cold environment to a room temp. Your thoughts/theories are appreciated. Fran Walker Histology Technical Specialist ETBH Knoxville _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From settembr <@t> umdnj.edu Fri Sep 3 10:53:38 2004 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Pre-cut controls Message-ID: I am with Karen. We pre-cut our controls. All of them and they always work well. We once had an inspector ask how long we keep them, which prompted me to keep the amount down to a months worth. Dana Settembre University Hospital - UMDNJ Newark, NJ USA >>> "Bauer, Karen" 9/3/2004 10:55:03 AM >>> We cut all of our controls ahead of time and have had no problems. Controls are tested before we use them and then of course are monitored when we actually use them. As far as I know, the Pathologists are happy with what we have and I have not had to change our way of handling controls. After we cut the controls, we put them in slide boxes and leave them open to air dry for a few hours or even overnight if cut late in the afternoon. We then place them in the cupboard and then use them when we need them. Good Luck to you... Karen Bauer HT(ASCP) Histology Supervisor Department of Pathology Luther Hospital Eau Claire, WI -----Original Message----- From: Joanne Mauger [mailto:MAUGER@email.chop.edu] Sent: Friday, September 03, 2004 9:40 AM To: flemons@bhset.org; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Pre-cut controls Fran, Some antigenicity can be lost when cut slides are stored for a long time. Time varies for different antigens, and some are not effected at all. Some people think refrigeration, or even freezing controls slides lengthens storage time. I personally think that it's best to cut what you will use in a few months time. Trial and error. Jo >>> "Fran Lemons" 09/03/04 10:15AM >>> Does anyone know why IHC control slides cannot be cut ahead of time? I've done it at other facilities, but I have been told it is not permissible here, without any explanation. Any ideas? Also, has anyone ever heard of refrigerating pre-cut IHC control slides? One person suggested to me that they stopped cutting them ahead of time because they were getting condensation on them & the stains wouldn't work. Then she informed me that they were stored in the fridge. Seems to me the condensation would have formed upon removing the slides from a cold environment to a room temp. Your thoughts/theories are appreciated. Fran Walker Histology Technical Specialist ETBH Knoxville _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Fri Sep 3 11:10:10 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Pre-cut controls Message-ID: We pre-cut all of our control tissue slides and store them at RT. We cut tonsil every week. Most others are used up in a month's time, but some do sit for a year or longer. Obviously, every laboratory is different, but you should do a study to determine how long your control slides remain reactive. 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 >>> "Fran Lemons" 09/03/04 10:15AM >>> Does anyone know why IHC control slides cannot be cut ahead of time? I've done it at other facilities, but I have been told it is not permissible here, without any explanation. Any ideas? Also, has anyone ever heard of refrigerating pre-cut IHC control slides? One person suggested to me that they stopped cutting them ahead of time because they were getting condensation on them & the stains wouldn't work. Then she informed me that they were stored in the fridge. Seems to me the condensation would have formed upon removing the slides from a cold environment to a room temp. Your thoughts/theories are appreciated. Fran Walker Histology Technical Specialist ETBH Knoxville _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Fri Sep 3 11:22:32 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Pre-cut controls Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA2D94C45@usca0082k08.labvision.apogent.com> Frank, Below is an abstract from an recent interesting paper on preservation of microarrays. Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm Lab Invest. 2004 Aug;84(8):1071-8. Long-term preservation of antigenicity on tissue microarrays. DiVito KA, Charette LA, Rimm DL, Camp RL. Department of Pathology, Yale University, New Haven, CT, USA. Tissue microarrays have facilitated the evaluation of large cohort studies; however, there is little data on the best method for preserving sections once they are cut. We assessed three methods of storing precut breast cancer microarray slides: paraffin coating and storage in a nitrogen desiccator, either alone or in combination. We tested the durability of three antigens, cytokeratin, estrogen receptor, and Ki-67 on microarrays stored under these conditions for 3 months at room temperature. Staining was assessed with both manual scoring using traditional brown stain (0-3+) as well as automated scoring using fluorescently stained sections. Staining intensity was compared to that obtained from freshly cut slides. Slides stored under ambient conditions (room temperature and air) for 3 months exhibited marked degradation of all target antigens, in some cases resulting in slides that were virtually unreadable. We found that combined paraffin coating and nitrogen storage resulted in the best preservation of antigenicity, with retention of 72-99% of the antigenicity of a freshly cut slide, depending upon the marker and detection system used. The use of either paraffin coating or nitrogen storage alone protected slides to a lesser degree. PMID: 15195116 [PubMed - indexed for MEDLINE] -----Original Message----- From: Fran Lemons [mailto:flemons@bhset.org] Sent: Friday, September 03, 2004 7:15 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Pre-cut controls Does anyone know why IHC control slides cannot be cut ahead of time? I've done it at other facilities, but I have been told it is not permissible here, without any explanation. Any ideas? Also, has anyone ever heard of refrigerating pre-cut IHC control slides? One person suggested to me that they stopped cutting them ahead of time because they were getting condensation on them & the stains wouldn't work. Then she informed me that they were stored in the fridge. Seems to me the condensation would have formed upon removing the slides from a cold environment to a room temp. Your thoughts/theories are appreciated. Fran Walker Histology Technical Specialist ETBH Knoxville _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Fri Sep 3 11:45:24 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Chromogranin B Message-ID: Happy Friday netters, my pathologists have me on a search for chromogranin B. I found clone PE-11 from Austria, but does anyone know of another source? Thanks Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX From David.Edmondson <@t> christie-tr.nwest.nhs.uk Fri Sep 3 11:44:36 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] RE: DAKO or VECTOR Message-ID: Think we have tried theirs, but ages ago and no advantage found. so we kept with what we knew best. Enjoy the weekend Dave I hear that there is an archeological dig on nearby . And open day over the weekend. So I shall maybe have a look for histological remains. ( Did they not use dyes and pigments back then.) www.marple-uk.com -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 03 September 2004 15:15 To: Edmondson David (RBV) NHS Christie Tr Subject: RE: DAKO or VECTOR Thank you, I am better informed now. Has this worked as well as VECTORS ABC kit? or have you tried the latter? Gayle Callis At 03:39 AM 9/3/2004, you wrote: >K0377, is a kit where a drop of HRP and a drop of Streptavidin get mixed >together in buffer, The data sheet that I have from 1997 makes no mention >of lifespan, now that I look >Dave > >-----Original Message----- >From: Gayle Callis [mailto:gcallis@montana.edu] >Sent: 02 September 2004 20:57 >To: Edmondson David (RBV) NHS Christie Tr >Subject: DAKO or VECTOR > > >Does DAKO have an avidin/bioin complex kit, I thought they has Labelled >Strepavidin aka LSAB? The ABC kit I am aware of is VECTOR's. > >At 10:47 AM 9/2/2004, you wrote: > >To Cao Xudong/co Histonet > >The DAKO ABC kit we used after several days, I thought they suggested a >week > >by way of a use-by time. But that is a few years back now. > > > >Dave > >Christie > >Manchester UK > >-----Original Message----- > >From: John C. Dennis [mailto:dennijc@vetmed.auburn.edu] > >Sent: 02 September 2004 15:54 > >To: Cao, Xudong > >Cc: histonet@lists.utsouthwestern.edu > >Subject: Re: [Histonet] vector ABC > > > > > > > >The Vector tech folk once told me that they'd used it up to several hours > >(8-12). I've kept it up to 4 hours but after that I dump it. > > > >John Carroll Dennis > >Anatomy, Physiology, and Pharmacology > >109 Greene Hall > >Auburn University, AL 36849 > > > > > >On Wed, 1 Sep 2004, Cao, Xudong wrote: > > > > > Dear all, > > > > > > does anybody know how long a ready-to-use ABC solution is good for? I am > >thinking about re-use the mixed solution if it is good for more than 48 > >hours... > > > > > > thanks. > > > > > > Xudong Cao > > > post-doc fellow > > > Brown 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 > > > >_______________________________________________ > >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 Fri Sep 3 11:59:18 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Magnification In-Reply-To: References: Message-ID: <6.0.0.22.1.20040903102840.01b0da88@gemini.msu.montana.edu> There are magnifiers that attach to your safety glasses or directly to eyeglasses. - the latter clip onto top of my prescription glasses. There are magnifier pasties you can attach directly to outside of safety glasses. Henry Schein company (they have a website) in dental products, has clip on eye loupes. I have also purchased them from knitting stores (Patternworks website, and sewing stores). The magnifiers that clip onto top of frames are more versatile and fit onto all eyeglasses - the ones with wires only fit onto large frames. These will run about $20/pair. There is also the possibilty that cheap magnifying eyewear, funky colors, etc can be worn under safety glasses - each technician can have their own eyeglasses for their favorite style and magnification. Cheap reading glasses have magnifications from 1, 1.25, 1.5, 1.75, 2 and 3X. They range in price from $10 to 20$ - found at WalMart, KMart, Target, supermarkets and drugstores, Pearle Vision . There are pencil thin reading glasses in mall kiosks, etc - up to 1.75 mag. These are totally funky, and coolest of all!! or At 08:48 AM 9/3/2004, you wrote: >Our Derm Path doc wants me to poll the Histonet subscribers about what >everyone uses when they're embedding tiny tissues such as derms. Any >creative magnifiers out there? > >Angela Bitting, HT(ASCP) >Technical Specialist, Histology >Geisinger Medical Center >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 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 convmcm <@t> cc.usu.edu Fri Sep 3 12:12:54 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Warthin Starey In-Reply-To: Message-ID: <000e01c491d9$40b37510$4a737b81@Cygnus> We use the same procedure listed below except that I dilute ALL of the reagents (including the Silver solutions) in acidulated water. I use Citric acid to make the DI water acidic. This is not done very scientifically --- I put a few grains of citric acid in about 1 liter of water. Check the pH and add citric acid until I get pH 4.0. After deparaffinizing, sections are well rinsed in DI water straight from the tap, then held in the acidulated water for 5 - 10 minutes. If I'm too busy to do the stain right away, I like to hold them in the acidulated water until I'm ready. I have found that using acidulated water is the key to a good WS stain... at least in this lab it is. Also, keep your solutions fresh. I like to prepare mine every 6 months. Happy staining *g* 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 Mildred Fail Sent: Thursday, September 02, 2004 4:48 AM To: histonet@lists.utsouthwestern.edu; Sarah.Blachford@ngh.nhs.uk Subject: Re: [Histonet] Warthin Starey Sarah, We use the following method with good results though the Steiner is used more often some of our pathologists prefer the Warthin-Starry. Solutions needed 1% Silver Nitrate 2% Silver Nitrate 0.15% Hydroquinone 5% Gelatin (use gentle heat to mix) All of the above are stored at 2-8 degrees Celsius for up to 6 months. !. Preheat a coplin jar with 1% silver nitrate in a 54-60 degree water bath for 30 minutes. Place your gelatin bottle in the water bath at the same time. 2. Deparaffinize and hydrate slides to distilled water 3. Place slides in the preheated 1% Silver Nitrate and incubate for 30 minutes in a 54-60 degree water bath. At the same time place a vial of 1.5 mls of 2% silver nitrate, a vial of 3.75 5% gelatin, and a vial of 2 mls of 0.15% hydroquinone into the 54-60 degree waterbath to preheat.for 30 minutes. 4. Prepare the developer solution by mixing the solutions in the following order. 1.5 mls of 2% silver Nitrate 3.75mls of 5 % gelatin and 2.0 mls of 0.15% hydroquinone 5. Remove slides from heated silver (DO NOT RINSE) Place slides horizontally on a paper towel and cover with developer. Allow sections to develop until they are a light golden brown. Check microscopically for end point 6. Rinse thoroughly in hot tap water 7. Dehydrate, clear and mount Rena Fail Medical University of SC >>> "Blachford, Sarah - Histopath Main" 09/02/04 05:27AM >>> Can anyone help us. Has anyone got a reliable warthin starey method. We have tried many methods but cannot get them to work. Also what is the use of gelatin in the method Sarah Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD This e-mail may contain confidential information and/or copyright material and is intended for the use of the addressee only. Any unauthorised use may be unlawful. The contents of this e-mail may be subject to public disclosure under the NHS Code of Openness or the Freedom of Information Act 2000. Unless legally exempt, the confidentiality of the message and your reply cannot be guaranteed. If you receive this e-mail by mistake, please advise the sender immediately. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From info <@t> instrumedics.com Fri Sep 3 12:14:21 2004 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] Magnification References: Message-ID: <005c01c491d9$74470a40$6401a8c0@INSTRUMEDICS22> Angela, Many dermatology laboratories use CryoGel embedding medium with the Gentle Jane Snap-Freezer(SAGJ) for preparing a skin block for frozen sectioning. Please visit our web site to see the details.www.instrumedics.com or contact me Bernice ----- Original Message ----- From: "Angela Bitting" To: Sent: Friday, September 03, 2004 10:48 AM Subject: [Histonet] Magnification Our Derm Path doc wants me to poll the Histonet subscribers about what everyone uses when they're embedding tiny tissues such as derms. Any creative magnifiers out there? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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 RITA.ANGEL <@t> UC.EDU Fri Sep 3 13:14:34 2004 From: RITA.ANGEL <@t> UC.EDU (Rita Angel) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] freezing mouse embryos Message-ID: <5.1.0.14.2.20040903140702.00b4be78@ucmail3.uc.edu> Hi all, I was wondering what is the preferred method for freezing mouse embryos. I have a researcher here that said he has previously fixed his embryos (13.5d) in 4% paraformaldehyde o/n then placed in 30% sucrose until the tissue sank. He then embedding in a mold with oct in liquid nitrogen. He said the morphology on these were very good. However, he brought some today that were not fixed, just frozen fresh in oct in liquid nitrogen. The morphology was very poor, and we were unable to see tissue structure very well. He thought it was because the tissue was not fixed before freezing. I'm wondering if perhaps the embryo was necrotic. I asked him if this was possible and he said he didn't think so. Has anyone done a comparison before? And has anyone had any problems with morphology when the embryos were just snap-frozen? Thank you for your help. I did find a few responses in the archives, but not very many. Rita Angel From Sue.Kapoor <@t> uhsi.org Fri Sep 3 14:24:46 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] HPV testing - hot/cold water Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F5659@khmcexch.uhsi.org> Hello histonet, I just found out that a pathologist can not run HPV testing unless the physician specifically requests it....can anyone tell why??? Also, I have a pathologist and a tech that use warm to hot water for the rinse steps when H&E staining a frozen section slide. I've always used cold water, I'm worried warm/hot water might encourage tissue to float off the slide. What does everyone else do? thanks, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From BlazekL <@t> childrensdayton.org Fri Sep 3 14:38:42 2004 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:23:59 2005 Subject: [Histonet] HPV testing - hot/cold water Message-ID: I've always used warm water when staining H&E on frozens. I've never had a problem with them floating off. I do use polylysine slides though. Linda Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 1 Children's Plaza Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org From Krat18 <@t> aol.com Fri Sep 3 15:47:41 2004 From: Krat18 <@t> aol.com (Krat18@aol.com) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Paraffin all over my floor....... Message-ID: <7e.578aef05.2e6a326d@aol.com> I found a long metal scraper, straight wooden handle, (not expensive) at Home Depot ..... the blade is just about 6" wide. We've used it for years. Karen_Raterman@ssmhc.com St. Mary's Health Center From pruegg <@t> ihctech.net Fri Sep 3 17:05:34 2004 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] muscle actin Message-ID: Has anyone used Dako's Muscle Actin HHF35 Code no. M0635 on mouse tissue? I realize that I will have to use m on m detection, I am just curious about this working on mouse or not. If you have this optimized please share your procedure with me including pretreatment if any for ffpe mouse lungs. Thank you, Patsy Ruegg From pruegg <@t> ihctech.net Fri Sep 3 17:06:22 2004 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] injecting tracers In-Reply-To: <6.1.0.6.2.20040830110655.024a0a08@tigger.uic.edu> Message-ID: Try a comany called Molecular Probes. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Lu Leach Sent: Monday, August 30, 2004 9:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] injecting tracers Dear Histonetters, I need advice on a fluorescent tracer to use for mouse injections that will hopefully survive tissue processing. If anyone has experience and wisdom in this area, please contact me. Thank you, Lu Leach University of Illinois at Chicago lleach@uic.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Sep 3 17:06:30 2004 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] alkaline phosphatase staining In-Reply-To: <1E0CC447E59C974CA5C7160D2A2854EC03C40E75@SJMEMXMB04.stjude.sjcrh.local> Message-ID: the enzyme alk.phos. stain for osteoblasts will not work on decalcified ffpe tissues in my experience. it will work limitedly on bone processed into GMA without decal and cold methanol fixed. I hear that there are antibodies to alk.phos. but I have not used them yet, has anyone else? Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Garlits, John Sent: Monday, August 30, 2004 11:04 AM To: HistoNet Server Subject: [Histonet] alkaline phosphatase staining Hello, I am interested in doing some alkaline phosphatase staining in decalcified, paraffin-fixed mouse long bone sections. Does anyone have any experience with alk phos staining, who could recommend a protocol or even a commercial kit? Thank you! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 Phone: 901-495-3961 Fax: 901-495-2176 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Sep 3 17:06:45 2004 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Pre-cut controls In-Reply-To: Message-ID: I store all my precious blocks for IHC controls at 4dc and any pre-cut slides are also stored dessicated at 4dc. There are some antibodies where it doesn't matter and some that are compromised if pre-cut and left at rt for long periods of time so I don't take a chance, but I am in research and have the space to store my tissues at 4dc. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Fran Lemons Sent: Friday, September 03, 2004 7:15 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Pre-cut controls Does anyone know why IHC control slides cannot be cut ahead of time? I've done it at other facilities, but I have been told it is not permissible here, without any explanation. Any ideas? Also, has anyone ever heard of refrigerating pre-cut IHC control slides? One person suggested to me that they stopped cutting them ahead of time because they were getting condensation on them & the stains wouldn't work. Then she informed me that they were stored in the fridge. Seems to me the condensation would have formed upon removing the slides from a cold environment to a room temp. Your thoughts/theories are appreciated. Fran Walker Histology Technical Specialist ETBH Knoxville _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jbaez <@t> interscopepath.com Fri Sep 3 17:35:55 2004 From: jbaez <@t> interscopepath.com (Baez, Janet) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] cryostats Message-ID: <9E956D8FEB06C2408B08AC16498325E9013994@scopemx1.interscope.com> Any feed back on the new cryostats from Sakura, Lecia and Thermo Shandon? Any comments would be greatly appreciated . Thanks. From Gervaip <@t> aol.com Fri Sep 3 18:10:23 2004 From: Gervaip <@t> aol.com (Gervaip@aol.com) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] HPV testing - hot/cold water Message-ID: <9b.4cb8ddb1.2e6a53df@aol.com> Sue, it is against the federal law (Medicaid and Medicare) to perform any tests, without a physician's written request in the pathology lab (clinical and anatomical). All the original request slips are kept for about 7 years. And we use cold tap water on frozens. Just never thought of using hot water. Do they use hot water to rinse the freezing compound off easier? Why do they say they use hot water? From Gervaip <@t> aol.com Fri Sep 3 18:12:58 2004 From: Gervaip <@t> aol.com (Gervaip@aol.com) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Gwen Deville of Alexandria, Louisiana ???? Message-ID: Does anyone know how I can get in touch with Gwen Deville. Must talk with her. From DayDawning <@t> wideopenwest.com Sat Sep 4 17:11:13 2004 From: DayDawning <@t> wideopenwest.com (Dawn Truscott) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] (no subject) Message-ID: <001001c492cc$17e0ab00$6401a8c0@wowway.com> Don't leave out the new Microm Cryostats from Richard-Allan!! Look for them in Toronto or at: www.rallansci.com Dawn Truscott, HT(ASCP) Sr. Territory Account Manager Richard-Allan Scientific From carl.hobbs <@t> kcl.ac.uk Mon Sep 6 03:00:26 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] re freezing mouse embryos Message-ID: <000d01c493e7$9351da40$e8345c9f@Carlos> IMHO, the morphology/histology of sections of unfixed, frozen embryos ( even after fixing the sections after cutting)is inferior to that of fixed, then frozen, tisssues, but should still be easily recognisable. Either the tissue is necrotic or that the freezing process is inadequate, leading to ice-crystal artefact. I do fixed and unfixed embryos by the processes you describe. What age are your embryos? Do you post-fix the sections? What in? Best wishes From carl.hobbs <@t> kcl.ac.uk Mon Sep 6 03:22:01 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] re muscle actin Message-ID: <001701c493ea$95de2310$e8345c9f@Carlos> You shouldn't have any problems.....provided that they are not inflammed. I've used that AB successfully on mouse etc ( pwax HEIR with citric pH6 or TRIS pH10). Dilution factor around 1/5000, when I use Goat anti mouse, biotinylated and StreptABC- Px , Dako. Using their recommended dilutions . Std immuno procedure using BSA solution to preblock and dilute the primary and secondary Abs. It also works on the pwax sections without HEIR, but not as strong, IMHO. I'll dig out a pic, if you wish From Mandy <@t> serotec.co.uk Mon Sep 6 03:25:11 2004 From: Mandy <@t> serotec.co.uk (Mandy Townsend) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] muscle actin Message-ID: Serotec are able to supply this antibody clone and the following reference uses the antibody in mouse tissues. Tsubura, A. et al. (1991). Immunophenotypic difference of Keratin expression in normal mammary glandular cells from five different species. Acta Anat. 140: 287 - 293. 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: www.serotec.com Serotec-Your first choice for antibodies! 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: pruegg@ihctech.net [mailto:pruegg@ihctech.net] Sent: Friday, September 03, 2004 11:06 PM To: histonet@pathology.swmed.edu Subject: [Histonet] muscle actin Has anyone used Dako's Muscle Actin HHF35 Code no. M0635 on mouse tissue? I realize that I will have to use m on m detection, I am just curious about this working on mouse or not. If you have this optimized please share your procedure with me including pretreatment if any for ffpe mouse lungs. Thank you, Patsy Ruegg _______________________________________________ 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 Internet. 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 rentonlf <@t> bru.wits.ac.za Mon Sep 6 03:24:20 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] blade sign Message-ID: <1094459060.911b91a0rentonlf@bru.wits.ac.za> Jason, bad batches of blades DO occur, although it would seem not all that frequently. It is hard to believe that ALL blocks at your facility contain hard inclusions (unless you are specifically dealing with bone & calcified tissue). Other things that might contribute are the way you treat your blade - wiping it with a Kimwipe before cutting for example, or touching it with forceps when picking up sections. Lastly, it may be that there is a contaminant in the wax, dust from nearby renovations might become incorporated in the embedding wax, thus giving rise to scratched & torn sections. Best regards -----Original Message----- From: "jason m" To: histonet@pathology.swmed.edu Date: Thu, 02 Sep 2004 11:34:40 -0700 Subject: [Histonet] blade sign anyone have tips for minimizing blade sign? i start at one end of fresh blade and work to the other end before throwing out the disposable blade. sometimes however, blade sign appears even with a new blade. is it just that some blocks have hard spots that nick the blade? thanks. J. _________________________________________________________________ Take advantage of powerful junk e-mail filters built on patented Microsoft SmartScreen Technology. http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines Start enjoying all the benefits of MSN Premium right now and get the first two months FREE*. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From abright <@t> brightinstruments.com Mon Sep 6 04:01:46 2004 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] blade sign Message-ID: Dear Jason, The problem you are encountering with disposable blades may have some similarities with this scenario of a story I heard. A large screw manufacturer in the UK had a problem on their production line with missing the slot on the heads of thousands of screws, they informed the work force that there would be a prize for the best idea for using these screws with no loss to the company. The person who won this prize suggested placing 2 unslotted screws in each pack of 1000 screws, as when the customers found the faulty screws they would just throw them away. As a manufacturer using many screws I know we throw out duff screws on a regular basis. 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: jason m [mailto:kosmicdog@hotmail.com] Sent: 02 September 2004 19:35 To: histonet@pathology.swmed.edu Subject: [Histonet] blade sign anyone have tips for minimizing blade sign? i start at one end of fresh blade and work to the other end before throwing out the disposable blade. sometimes however, blade sign appears even with a new blade. is it just that some blocks have hard spots that nick the blade? thanks. J. _________________________________________________________________ Take advantage of powerful junk e-mail filters built on patented Microsoft(r) SmartScreen Technology. http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU =http://hotmail.com/enca&HL=Market_MSNIS_Taglines Start enjoying all the benefits of MSN(r) Premium right now and get the first two months FREE*. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ctsblack <@t> capeheart.uct.ac.za Mon Sep 6 05:31:42 2004 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Anti-Rat Vimentin/Granulocyte/CD 3 Message-ID: Hi There Has anybody established Vimentin/Granulocyte and CD 3 markers on Rat tissue yet, and if so, can you share the adaptions etc please??? Many Thanks Melanie. -- Cardiovascular Research Unit Div. of Cardiothoracic Surgery Chris Barnard Building University of Cape Town Anzio Road Observatory 7925 Republic of South Africa Tel +27 21 406-6589 Cel +27 82 469-3352 Fax +27 21 448-5935 From Mandy <@t> serotec.co.uk Mon Sep 6 05:47:10 2004 From: Mandy <@t> serotec.co.uk (Mandy Townsend) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Anti-Rat Vimentin/Granulocyte/CD 3 Message-ID: Serotec are able to supply antibodies to rat CD3 and Vimentin cross reactive with rat, which are suitable for use on both cryostat and paraffin sections (using HIER). I have attached a link to the datasheets should this be of assistance. http://www.serotec.com/asp/datasheet.asp?code=MCA772 http://www.serotec.com/asp/datasheet.asp?code=MCA862 http://www.serotec.com/asp/datasheet.asp?code=MCA862HT (sample size) We also are able to supply antibodies to rat granulocytes, but unfortunately, these have not been tested in paraffin sections only cryostat. http://www.serotec.com/asp/datasheet.asp?code=MCA967 http://www.serotec.com/asp/datasheet.asp?code=MCA149 If you require any further information, please do not hesitate to contact me. 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: www.serotec.com Serotec-Your first choice for antibodies! 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: Melanie Black [mailto:ctsblack@capeheart.uct.ac.za] Sent: Monday, September 06, 2004 11:32 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Anti-Rat Vimentin/Granulocyte/CD 3 Hi There Has anybody established Vimentin/Granulocyte and CD 3 markers on Rat tissue yet, and if so, can you share the adaptions etc please??? Many Thanks Melanie. -- Cardiovascular Research Unit Div. of Cardiothoracic Surgery Chris Barnard Building University of Cape Town Anzio Road Observatory 7925 Republic of South Africa Tel +27 21 406-6589 Cel +27 82 469-3352 Fax +27 21 448-5935 _______________________________________________ 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 Joanne.Hosker <@t> mail.bhrv.nwest.nhs.uk Mon Sep 6 08:08:17 2004 From: Joanne.Hosker <@t> mail.bhrv.nwest.nhs.uk (Joanne Hosker) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] working in canada[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D0428C3A4@BHRV_NT_11> Does anyone out there have any information on temporary vacancies in Vancouver or Toronto?? Or info on any agencies I can contact in these cities?? I am going to be looking for a job in histology from Dec 2004. Thanks Joanne Hosker From John.Auld <@t> whnt.nhs.uk Mon Sep 6 12:02:45 2004 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Re: Temperatures for IHC Message-ID: Ronald Be careful using higher temperatures. 2 - 3 years ago I did a study using 6 Abs at different temperatures as well as asking on Histonet. Some Abs, ER & PR particularly, give weaker staining as well as staining fewer cells at temperatures above 25 C. If you want to stain at a higher temp I suggest validating every Ab before hand. Cheers John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral Internal extn 2560 External Tel 0151 604 7025 Message: 7 Date: Wed, 1 Sep 2004 14:24:34 -0400 From: "Ronald P. Wilson" Subject: [Histonet] Temperatures for IHC To: "Histonet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" We are in the process of optimizing our IHC procedures. We have switched to using some of the reagents from Phoenix Biotechnologies. Their recommended protocols suggest primary and secondary antibody as well as strepavidin and chromagen incubations be done at 55?C for much shorter time periods (4-10 minutes). I have also heard that some labs do their incubations at 37?C. Is it common practice to use temperatures higher than room temperature? Are there advantages other than just shortening the incubation time? If anyone is using the higher temperature, how do you do this for bench-top procedures? I know that some of the staining stations such as Microprobe have built-in incubation chambers to do this. Ronald P. Wilson, V.M.D., M.S. Associate Professor Department of Comparative Medicine, HO54 Penn State University College of Medicine M. S. Hershey Medical Center 500 University Drive Hershey, PA 17033 phone: 717-531-8460 fax: 717-531-5001 e-mail: rpw4@psu.edu From laurie.reilly <@t> jcu.edu.au Mon Sep 6 19:10:30 2004 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] p53 staining Message-ID: <5.1.0.14.0.20040907092200.00a39ec0@mail.jcu.edu.au> Dear All, Does anyone have a protocol for doing p53 staining on Formalin Fixed Paraffin Embedded tissues. We are trying various methods of antigen retrieval and incubation times on canine mammary tumour, and hope to be able to apply the technique to canine mast cell tumours. Thanks for your consideration. Regards, Laurie. Mr.Laurie Reilly Ph 07 4781 4468 Physiology & Pharmacology Fax 07 4779 1526 Aust.Inst.of Tropical Vet.& Animal Sc. James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. From conniegrubaugh <@t> hotmail.com Mon Sep 6 19:10:11 2004 From: conniegrubaugh <@t> hotmail.com (connie grubaugh) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Nevada Society of Histotechnology Message-ID: The program for the NVSH upcoming symposium in Las Vegas October 29 and 30th. Friday: at 6pm Dr. Steve Ruhoy- Cat Scratch Disease to follow Dr. Steve Kolker- Leprosy the clinical and histological diagnosis. Saturday 8am Dr. La Vent Keskintepe The technical process of invitro fertilization of the human embryo. 10am Ms Lindsay Sinn AAPA - The Silver Bullet Interesting forensic cases. 1pm Dr. Gary Telgenhoff-Clark County Coroner's Office Forensic Pathologist 3 pm Dr. Stacey Garry - African histomorphology If anyone would like more information email me and I will send you a program by mail or fax. Connie Grubaugh _________________________________________________________________ [1]Express yourself instantly with MSN Messenger! Download today - it's FREE! References 1. http://g.msn.com/8HMAENUS/2734??PS=47575 From Sarah.Blachford <@t> ngh.nhs.uk Tue Sep 7 04:08:34 2004 From: Sarah.Blachford <@t> ngh.nhs.uk (Blachford, Sarah - Histopath Main) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] gold standard Message-ID: <854FA06D8BE2D611ABA30030052D1DF6FA58F3@NGH_EXCHANGE1> can any one tell me why the gold standard is IMF for the blistering skin conditions such as Dermatitis herpetiformis, pemphigoid Sarah Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD This e-mail may contain confidential information and/or copyright material and is intended for the use of the addressee only. Any unauthorised use may be unlawful. The contents of this e-mail may be subject to public disclosure under the NHS Code of Openness or the Freedom of Information Act 2000. Unless legally exempt, the confidentiality of the message and your reply cannot be guaranteed. If you receive this e-mail by mistake, please advise the sender immediately. Thank you. From pstefanova <@t> sten.sunnybrook.utoronto.ca Tue Sep 7 08:11:54 2004 From: pstefanova <@t> sten.sunnybrook.utoronto.ca (Petia P Stefanova) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] working in canada[Scanned] References: <1030B679AD69D6119C3F00080210DD9D0428C3A4@BHRV_NT_11> Message-ID: <004301c494dc$3f2d5cf0$9d194c8e@WS21203> Hi Joanne, See this link http://www.workopolis.com/index.html then on the upper left side and you 'll see different categories. Go to Healthcare and you'll find positions and employment agencies as well. This is an employment agency http://www.kellyscientificresources.com Good luck! Petia ----- Original Message ----- From: "Joanne Hosker" To: Sent: Monday, September 06, 2004 9:08 AM Subject: [Histonet] working in canada[Scanned] > Does anyone out there have any information on temporary vacancies in > Vancouver or Toronto?? Or info on any agencies I can contact in these > cities?? I am going to be looking for a job in histology from Dec 2004. > > > > Thanks > > Joanne Hosker > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gcallis <@t> montana.edu Tue Sep 7 09:32:43 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Paraffin on floors In-Reply-To: <7e.578aef05.2e6a326d@aol.com> References: <7e.578aef05.2e6a326d@aol.com> Message-ID: <6.0.0.22.1.20040907082932.01b1b2f8@gemini.msu.montana.edu> We have used the industrial type rugs , seen at entrances of buildings, and place these at microtomes and embedding areas. They are vacuumed to remove paraffin trimmings. We had to go to rugs as our floors became so slippery, safety was an issue - particularly after a visiting researcher slipped, went backwards and if he hadn't caught a doorknob on way down, would have cracked his skull on a concrete floor At 02:47 PM 9/3/2004, you wrote: >I found a long metal scraper, straight wooden handle, (not expensive) at Home >Depot ..... the blade is just about 6" wide. We've used it for years. > >Karen_Raterman@ssmhc.com >St. Mary's Health Center >_______________________________________________ >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 Sep 7 09:41:38 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] blade sign In-Reply-To: <1094459060.911b91a0rentonlf@bru.wits.ac.za> References: <1094459060.911b91a0rentonlf@bru.wits.ac.za> Message-ID: <6.0.0.22.1.20040907083636.01adf4d0@gemini.msu.montana.edu> Another source of dirt, etc is from the paraffin dispenser. We have experienced tissues infiltrated and embedded in paraffin from another lab (of which there is nothing wrong with paraffin from a reliable manufacturer) but tissues seemed to have "sand" and sections were a nightmare. Ribbon looked like a spaghetti factory production. After reembedding in another clean paraffin, the sand syndrome disappeared - suspect was a dirty dispenser and failure to stir the paraffin BEFORE embedding to redistribute plastic polymers and additives evenly in the melted paraffin mixture. At 02:24 AM 9/6/2004, you wrote: >Jason, > >bad batches of blades DO occur, although it would seem not all that >frequently. It is hard to believe that ALL blocks at your >facility contain hard inclusions (unless you are specifically dealing >with bone & calcified tissue). >Other things that might contribute are the way you treat your blade - >wiping it with a Kimwipe before cutting for example, or touching it with >forceps when picking up sections. Lastly, it may be that there is a >contaminant in the wax, dust from nearby renovations might become >incorporated in the embedding wax, thus giving rise to scratched & torn >sections. > >Best regards > > > > >-----Original Message----- >From: "jason m" >To: histonet@pathology.swmed.edu >Date: Thu, 02 Sep 2004 11:34:40 -0700 >Subject: [Histonet] blade sign > >anyone have tips for minimizing blade sign? i start at one end of fresh >blade and work to the other end before throwing out the disposable blade. >sometimes however, blade sign appears even with a new blade. is it just that >some blocks have hard spots that nick the blade? > >thanks. >J. > >_________________________________________________________________ >Take advantage of powerful junk e-mail filters built on patented Microsoft >SmartScreen Technology. >http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines > > Start enjoying all the benefits of MSN Premium right now and get the >first two months FREE*. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >Louise Renton >Bone Research Unit >University of the Witwatersrand >Johannesburg >South Africa >.......so what IS the speed of dark? > >_______________________________________________ >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 gliuygao <@t> hotmail.com Tue Sep 7 09:42:32 2004 From: gliuygao <@t> hotmail.com (yan gao) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] (no subject) Message-ID: Can someone tell me how to automatically reply an E-mail when I am not in my office? Such as I am not in my office, I shall be return to my office at 15 September 2004. I know this is not a histology question but can someone nicely tell me. I am very appreciated. Yan _________________________________________________________________ [1]FREE pop-up blocking with the new MSN Toolbar get it now! References 1. http://g.msn.com/8HMBENUS/2752??PS=47575 From STEGTM <@t> samcstl.org Tue Sep 7 09:42:53 2004 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] I have a funny Message-ID: I agree. As we are all aware, this site is monitored by vendors/salespeople, and what better reason for them to monitor such than being able to get feedback from working histotechs about the products they represent. This does not necessarily mean that the "net" would be used to 'intimidate', but since we're aware that these reps are privy to discussions of goods and services, good and bad, between histotechs, why not let them know the truth? Peace, Terre >>> "Lucy Brooks" 9/1/2004 10:23:11 AM >>> I personally hope the histonet isn't used to "intimidate companies" , but I do feel that if you have a problem and the company who you have that problem with doesn't deal with it in a reasonable amount of time, it is nice to warn other people out their of what they can expect when dealing with these said companies. ----- Original Message ----- From: "Sarah Jones" To: ; ; Sent: Tuesday, August 31, 2004 6:56 PM Subject: Re: [Histonet] I have a funny > Ah, this begs an ethical question.... should the histonet be used as a > threat to intimidate companies? > Is this an Us against Them mentality? > Discuss amongst yourselves. (a little "Coffee Talk" humor there for > you Saturday Night Live fans) > Oye Vey..................... Sarah > > > Sarah Jones, HT(ASCP) > Histology Lab > Dept. of Veterinary Integrative Biosciences > College of Veterinary Medicine > Texas A&M University > College Station, TX 77843-4458 > phone: 979-845-3177 > fax: 979-458-3499 > > > >>> "Lucy Brooks" 8/31/2004 6:10:52 PM >>> > Joe!! > > Speak your mind!!! > The Histonet is a powerful thing!! > > ----- Original Message ----- > From: "Joe Nocito" > To: "Histonet" > Sent: Tuesday, August 31, 2004 4:01 PM > Subject: [Histonet] I have a funny > > > > Hello all, > > for once, I have, what I think is funny, is a short story. A couple > of > > months ago, I purchased a piece of equipment from a vendor (who will > remain > > anonymous, at this time at least). I was told it was 6 weeks for > delivery. > > It's been over 8 weeks. I called the rep and left a voice mail asking > when > I > > can expect my equipment. S/he called me right back. I asked when I > can > > expect the equipment. Then I said "Don't make me go on the > Histonet". > > My techs started laughing and I couldn't keep a straight face. The > rep was > > all apologetic and promised me I'll have my equipment in 2 weeks. > Gee, I > > didn't think I was getting that bad. Maybe I should settle down a > little > > bit. > > > > 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From algranth <@t> u.arizona.edu Tue Sep 7 10:13:21 2004 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Paraffin on floors In-Reply-To: <6.0.0.22.1.20040907082932.01b1b2f8@gemini.msu.montana.edu> References: <7e.578aef05.2e6a326d@aol.com> <7e.578aef05.2e6a326d@aol.com> Message-ID: <4.3.2.7.2.20040907080517.00c65968@algranth.inbox.email.arizona.edu> We just got floor mats from MarketLab that were designed for histology labs. Paraffin will fall into grooves in the mats and we can vacuum it up with the shop vac I also just got. I keep one in the embedding area and one near the processor and on the way out the door. The mats come in two sizes but you can special order any size you need. They work great. We also use those sticky mats (human fly paper) to catch paraffin that may stick to your shoes. Andi At 08:32 AM 9/7/2004 -0600, you wrote: >We have used the industrial type rugs , seen at entrances of buildings, >and place these at microtomes and embedding areas. They are vacuumed to >remove paraffin trimmings. We had to go to rugs as our floors became so >slippery, safety was an issue - particularly after a visiting researcher >slipped, went backwards and if he hadn't caught a doorknob on way down, >would have cracked his skull on a concrete floor > > > >At 02:47 PM 9/3/2004, you wrote: > >>I found a long metal scraper, straight wooden handle, (not expensive) at Home >>Depot ..... the blade is just about 6" wide. We've used it for years. >> >>Karen_Raterman@ssmhc.com >>St. Mary's Health Center >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : 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 JEllin <@t> yumaregional.org Tue Sep 7 10:54:47 2004 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] P-63 help please!! Message-ID: Currently we are in a dilema we are looking for a P-63 antibody that works well with thet Ventanna stainers. We use to buy it fomr cell marque and they are now discontinuing thier product. If any one can help with this it would be greatly appreciated. The clone that we are currently using is a mouse monoclonal clone (4A4). Thanks Jesus Ellin Yuma Regional Medical Center From David.Edmondson <@t> christie-tr.nwest.nhs.uk Tue Sep 7 11:08:34 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] P-63 help please!! Message-ID: -----Original Message----- From: Edmondson David (RBV) NHS Christie Tr Sent: 07 September 2004 17:08 To: 'Jesus Ellin' Subject: RE: [Histonet] P-63 help please!! DAKO Cat M7247 is the same clone, We are using it diluted on our Techmate 500 Dave Christie Manchester UK -----Original Message----- From: Jesus Ellin [mailto:JEllin@yumaregional.org] Sent: 07 September 2004 16:55 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] P-63 help please!! Currently we are in a dilema we are looking for a P-63 antibody that works well with thet Ventanna stainers. We use to buy it fomr cell marque and they are now discontinuing thier product. If any one can help with this it would be greatly appreciated. The clone that we are currently using is a mouse monoclonal clone (4A4). Thanks Jesus Ellin Yuma Regional Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DDittus787 <@t> aol.com Tue Sep 7 11:28:39 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Paraffin on floors Message-ID: <294973E3.1EE626C8.0A1F969F@aol.com> we just recently renovated histology and acquired an anti-parrafin floor, has rough texture for non slippage and wax beads up. dana From sa.drew <@t> hosp.wisc.edu Tue Sep 7 11:33:22 2004 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] P-63 help please!! Message-ID: We are currently using BioCare's p63 ( BC4A4) on our Ventana immunostainer's. I'm not sure if this is the same clone, but feel free to contact us on protocol details if you'd like... -----Original Message----- From: Jesus Ellin [mailto:JEllin@yumaregional.org] Sent: Tuesday, September 07, 2004 10:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] P-63 help please!! Currently we are in a dilema we are looking for a P-63 antibody that works well with thet Ventanna stainers. We use to buy it fomr cell marque and they are now discontinuing thier product. If any one can help with this it would be greatly appreciated. The clone that we are currently using is a mouse monoclonal clone (4A4). Thanks Jesus Ellin Yuma Regional Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From donna <@t> phxbio.com Tue Sep 7 12:00:59 2004 From: donna <@t> phxbio.com (Donna Brown) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] I have a funny In-Reply-To: References: Message-ID: <6.1.0.6.0.20040907115053.029eed28@phxbio.com> Thanks for stating this so well, Terre. My work includes tech support our company and it's extremely useful to me to know when one of our items is working well or if it's difficult to use. Most customers simply don't take the time to provide any feedback but it's important for the financial health of each company to provide the highest quality products and to fix problems promptly. The bottom line is that we're all in this business together- whether you're the person in the lab providing the services your hospital and/or doctor need, or if you're associated with a company that provides supplies, equipment, etc. We rely on one another; everyone benefits from the truth. Donna >I agree. As we are all aware, this site is monitored by >vendors/salespeople, and what better reason for them to monitor such >than being able to get feedback from working histotechs about the >products they represent. This does not necessarily mean that the "net" >would be used to 'intimidate', but since we're aware that these reps are >privy to discussions of goods and services, good and bad, between >histotechs, why not let them know the truth? Peace, Terre > > >>> "Lucy Brooks" 9/1/2004 10:23:11 AM >>> >I personally hope the histonet isn't used to "intimidate companies" , >but I >do feel that if you have a problem and the company who you have that >problem >with doesn't deal with it in a reasonable amount of time, it is nice to >warn >other people out their of what they can expect when dealing with these >said >companies. >----- Original Message ----- >From: "Sarah Jones" >To: ; ; > >Sent: Tuesday, August 31, 2004 6:56 PM >Subject: Re: [Histonet] I have a funny > > > > Ah, this begs an ethical question.... should the histonet be used as >a > > threat to intimidate companies? > > Is this an Us against Them mentality? > > Discuss amongst yourselves. (a little "Coffee Talk" humor there >for > > you Saturday Night Live fans) > > Oye Vey..................... Sarah > > > > > > Sarah Jones, HT(ASCP) > > Histology Lab > > Dept. of Veterinary Integrative Biosciences > > College of Veterinary Medicine > > Texas A&M University > > College Station, TX 77843-4458 > > phone: 979-845-3177 > > fax: 979-458-3499 > > > > > > >>> "Lucy Brooks" 8/31/2004 6:10:52 PM >>> > > Joe!! > > > > Speak your mind!!! > > The Histonet is a powerful thing!! > > > > ----- Original Message ----- > > From: "Joe Nocito" > > To: "Histonet" > > Sent: Tuesday, August 31, 2004 4:01 PM > > Subject: [Histonet] I have a funny > > > > > > > Hello all, > > > for once, I have, what I think is funny, is a short story. A >couple > > of > > > months ago, I purchased a piece of equipment from a vendor (who >will > > remain > > > anonymous, at this time at least). I was told it was 6 weeks for > > delivery. > > > It's been over 8 weeks. I called the rep and left a voice mail >asking > > when > > I > > > can expect my equipment. S/he called me right back. I asked when I > > can > > > expect the equipment. Then I said "Don't make me go on the > > Histonet". > > > My techs started laughing and I couldn't keep a straight face. The > > rep was > > > all apologetic and promised me I'll have my equipment in 2 weeks. > > Gee, I > > > didn't think I was getting that bad. Maybe I should settle down a > > little > > > bit. > > > > > > 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 > > >_______________________________________________ >Histonet mailing list >Histonet@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 lucyb <@t> biocare.net Tue Sep 7 12:06:46 2004 From: lucyb <@t> biocare.net (Lucy Brooks) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Chromogranin B References: Message-ID: <004801c494fd$0eaafc60$0a01a8c0@LUCYSALES> Joe, Go to this site, B-D Biosciences sells it. :) Hope this helps!!!! http://www.bdbiosciences.com/ptProduct.jsp?prodId=27014 ----- Original Message ----- From: "Joe Nocito" To: "Histonet" Sent: Friday, September 03, 2004 9:45 AM Subject: [Histonet] Chromogranin B > Happy Friday netters, > my pathologists have me on a search for chromogranin B. I found clone PE-11 > from Austria, but does anyone know of another source? 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 settembr <@t> umdnj.edu Tue Sep 7 13:37:45 2004 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] p53 staining Message-ID: Hello Laurie, I use p53 on human tissue. The antibody is from DakoCytomation used at a 1 :150 dilution with a colon cancer as a control. I use DakoCytomations Target Retrieval Solution in a steamer for 40 minutes Antibody is incubated at Room Temperature for 15 minutes then Dako's LSAB2 kit for detection. Hope this helps. Dana Settembre University Hospital - UMDNJ Newark, NJ USA >>> Laurie Reilly 9/6/2004 8:10:30 PM >>> Dear All, Does anyone have a protocol for doing p53 staining on Formalin Fixed Paraffin Embedded tissues. We are trying various methods of antigen retrieval and incubation times on canine mammary tumour, and hope to be able to apply the technique to canine mast cell tumours. Thanks for your consideration. Regards, Laurie. Mr.Laurie Reilly Ph 07 4781 4468 Physiology & Pharmacology Fax 07 4779 1526 Aust.Inst.of Tropical Vet.& Animal Sc. James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From weneng2004 <@t> yahoo.com Tue Sep 7 13:54:39 2004 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] plastic in paraffin block Message-ID: <20040907185439.815.qmail@web53405.mail.yahoo.com> Hi, I got some skin tissue in formalin that covered with plastic film. If I peel off plastic it will damaged the wound. Looks like I have to process them with the plastic on. But I worried that the plastic will give me big headache when I section them. Anybody has good idea? Thanks in advance! Wen --------------------------------- Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. From akbitting <@t> geisinger.edu Tue Sep 7 14:36:55 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Hp controls Message-ID: I'm looking for a supplier of good quality H. pylori controls. Any suggestions? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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 pruegg <@t> ihctech.net Tue Sep 7 15:37:49 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] (no subject) In-Reply-To: Message-ID: Please unsubscribe to histonet before you do that or we will have a bounce loop which will crash the list. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of yan gao Sent: Tuesday, September 07, 2004 7:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Can someone tell me how to automatically reply an E-mail when I am not in my office? Such as I am not in my office, I shall be return to my office at 15 September 2004. I know this is not a histology question but can someone nicely tell me. I am very appreciated. Yan _________________________________________________________________ [1]FREE pop-up blocking with the new MSN Toolbar get it now! References 1. http://g.msn.com/8HMBENUS/2752??PS=47575 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Tue Sep 7 15:54:22 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] (no subject) Message-ID: <83AACDB0810528418AA106F9AE9B7F7E278231@sjhaexc02.sjha.org> HOW THE LIST WORKS: To subscribe to the list or change your subscription to the daily digest mode etc. you need to go to http://lists.utsouthwestern.edu/mailman/listinfo/histonet You will need to request a password if you do not have one. You can go to the website above - where you subscribed to the list and unsubscribe while you are out of the office. Then, when you return, go back to this web site again and subscribe. Good luck! j:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of yan gao Sent: Tuesday, September 07, 2004 7:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Can someone tell me how to automatically reply an E-mail when I am not in my office? Such as I am not in my office, I shall be return to my office at 15 September 2004. I know this is not a histology question but can someone nicely tell me. I am very appreciated. Yan _________________________________________________________________ [1]FREE pop-up blocking with the new MSN Toolbar get it now! References 1. http://g.msn.com/8HMBENUS/2752??PS=47575 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From JNocito <@t> Pathreflab.com Tue Sep 7 16:14:14 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] P-63 help please!! In-Reply-To: Message-ID: Jesus, we have Biocare's p63 working on our XTs. We also have their PIN cocktail which includes p63 & p504s working. Their number is 1-800-799-9499 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 Jesus Ellin Sent: Tuesday, September 07, 2004 10:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] P-63 help please!! Currently we are in a dilema we are looking for a P-63 antibody that works well with thet Ventanna stainers. We use to buy it fomr cell marque and they are now discontinuing thier product. If any one can help with this it would be greatly appreciated. The clone that we are currently using is a mouse monoclonal clone (4A4). Thanks Jesus Ellin Yuma Regional Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From karrie.langdon <@t> sympatico.ca Tue Sep 7 20:18:54 2004 From: karrie.langdon <@t> sympatico.ca (Walker/Langdon) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Specificity of Masson's trichrome Message-ID: <005d01c49541$cf1ddc30$8a19fea9@walkercvzdlx4z> Does anyone have details regarding the specificity of the collagen staining when tissues(especially lung) are stained with Masson's trichrome? Is it possible that fibronectin and/or alpha smooth muscle actin can also be stained? We are trying to reconcile some IHC and in vitro data. Thanks so much! Carrie From rentonlf <@t> bru.wits.ac.za Wed Sep 8 03:11:43 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] plastic in paraffin block Message-ID: <1094631103.92068480rentonlf@bru.wits.ac.za> Hi Wen, You'll probably find that the wound dressing (sounds like an OpSite dressing) will peel away when exposed to solvent in processing. Fixation alone might be enough to harden the tissue and allow the dressing to be peeled away more easily without damaging the underlying skin. You could also try to get a sample from the surgeon/hospital/doctor where the tissue originated and do a little test in the lab first. After having tested numerous tape & dressing samples for totally another reason, I have found that most are more or less removed in either alcohol or xylene. Good luck -----Original Message----- From: wen eng To: histonet@lists.utsouthwestern.edu Date: Tue, 7 Sep 2004 11:54:39 -0700 (PDT) Subject: [Histonet] plastic in paraffin block Hi, I got some skin tissue in formalin that covered with plastic film. If I peel off plastic it will damaged the wound. Looks like I have to process them with the plastic on. But I worried that the plastic will give me big headache when I section them. Anybody has good idea? Thanks in advance! Wen --------------------------------- 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 Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From ctsblack <@t> capeheart.uct.ac.za Wed Sep 8 04:14:35 2004 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Double/Tripple Labelling - Fluorescent Message-ID: Hi There I know this topic has been around before, so forgive me!! I am attempting doulble and then tripple fluorescent labelling. Do I need to add a fixation step between Primary antibodies, to secure the first Ag/Ab complex??? Any help with this proceedure will be greatly appreciated. Many Thanks Melanie. -- Cardiovascular Research Unit Div. of Cardiothoracic Surgery Chris Barnard Building University of Cape Town Anzio Road Observatory 7925 Republic of South Africa Tel +27 21 406-6589 Cel +27 82 469-3352 Fax +27 21 448-5935 From andromeda_tm <@t> libero.it Wed Sep 8 05:48:05 2004 From: andromeda_tm <@t> libero.it (andromeda_tm) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Land snail dissection. Message-ID: Torino 08 September 2004 (ITALY) Hi all, I am an amateur naturalist. I like to study the histology of animal tissues by an optical microscope in transmitted light. I am interested to land (terrestrial) snails. I know for relaxing the snail to use a solution of 50mM of MgCl2 by an injection (2 ml.) into the foot. Could someone give me some detailed information how to proceed to the snail dissection? Thank you. With my Best Regards, Massimo From celebrej <@t> HHSC.CA Wed Sep 8 06:12:35 2004 From: celebrej <@t> HHSC.CA (Celebre Julia) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Tissue dye Message-ID: <3AADFB88753AD31189C100902786B91C0E41CEB2@hch_nt_exchange.hhsc.ca> Morning all. Can anyone tell me what they use to ink their tissues prior to processing? We need to find a second and third tissue dye marker for our Pathologist Assistant since our supplier has discontinued the ones we currently use. Any suggestions are greatly appreciated. Thanks Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca 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 akbitting <@t> geisinger.edu Wed Sep 8 07:52:51 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Modified acid fast stain for actinomyces Message-ID: Anyone have a recipe for this stain? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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 Terry.Marshall <@t> rothgen.nhs.uk Wed Sep 8 08:19:44 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Modified acid fast stain for actinomyces Message-ID: No modification is needed. Neither does it stain the Actinomyces. Rather, the club shaped ends of the filaments, being an antigen-antibody complex - known as Splendore-Hoeppli phenomenon - are stained red. They are not necessarily present. 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: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: 08 September 2004 13:53 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Modified acid fast stain for actinomyces Anyone have a recipe for this stain? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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 lab <@t> mcpathology.com Wed Sep 8 07:20:29 2004 From: lab <@t> mcpathology.com (Lab) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] GIARDIA CONTROL TISSUE Message-ID: <006f01c4959e$3ac4bd80$70cfa8c0@LAB> If anyone out there has a block or two of tissue for giardia we would be greatful if you could send us some.Maybe we can return the favor one day. Thanks, Amy Boan HT Marlboro-Chesterfield Pathology 672 Hwy 9 West Bennettsville, SC 29512 From gcallis <@t> montana.edu Wed Sep 8 09:56:07 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Land snail dissection. In-Reply-To: References: Message-ID: <6.0.0.22.1.20040908084729.01b378c0@gemini.msu.montana.edu> Becaue of the hard calcified shell, dissection was still difficult. The snail didn't always relax all the way out of the shell. I think we also used another way to relax snail, but do not recall what was used (many years ago!), as long as your method works, use it. Since we could never get a snail totally out of shell (they don't give it up easily!), we fixed snails whole, then decalcified them with shells intact, and processed them into paraffin. There is a "tooth" of some sort, I do not recall what my snail experts called it, but it can create problems during sectioning. It was very hard and caused section damage. There is always a possiblity that after you decalcify the shell, you can remove it very carefully to reach soft body parts. We had wonderful sections with thin shell intact - a total histological preparation. We decalcified in 10 to 15% formic acid after the snail was totally fixed. At 04:48 AM 9/8/2004, you wrote: >Torino 08 September 2004 >(ITALY) > > >Hi all, > >I am an amateur naturalist. >I like to study the histology of animal tissues by an optical microscope >in transmitted light. >I am interested to land (terrestrial) snails. >I know for relaxing the snail to use a solution of 50mM of MgCl2 by an >injection (2 ml.) into the foot. >Could someone give me some detailed information how to proceed to the >snail dissection? >Thank you. >With my Best Regards, > >Massimo 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 gu.lang <@t> gmx.at Wed Sep 8 11:15:06 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] plastic in paraffin block References: <20040907185439.815.qmail@web53405.mail.yahoo.com> Message-ID: <001f01c495bf$017ba8b0$eeeea8c0@server> Would be interesting, if the plastic will dissolve in the xylol? Gudrun ----- Original Message ----- From: "wen eng" To: Sent: Tuesday, September 07, 2004 8:54 PM Subject: [Histonet] plastic in paraffin block > Hi, > > I got some skin tissue in formalin that covered with plastic film. If I peel off plastic it will damaged the wound. Looks like I have to process them with the plastic on. But I worried that the plastic will give me big headache when I section them. > > Anybody has good idea? > > Thanks in advance! > > Wen > > > > --------------------------------- > 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 asmith <@t> mail.barry.edu Wed Sep 8 11:23:01 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Land snail dissection. Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3BDF@exchsrv01.barrynet.barry.edu> Many centuries ago, I forced a snail out of its shell by shredding a pack of cigarettes into a pint of water and dropping the snail into it. Borradaile's THE INVERTEBRATA has instructions for dissecting the European garden snail Helix pomatia (pp. 604-610 in the 4th edition). The book is out of print, but available used. 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 Gayle Callis Sent: Wednesday, September 08, 2004 10:56 AM To: andromeda_tm; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Land snail dissection. Becaue of the hard calcified shell, dissection was still difficult. The snail didn't always relax all the way out of the shell. I think we also used another way to relax snail, but do not recall what was used (many years ago!), as long as your method works, use it. Since we could never get a snail totally out of shell (they don't give it up easily!), we fixed snails whole, then decalcified them with shells intact, and processed them into paraffin. There is a "tooth" of some sort, I do not recall what my snail experts called it, but it can create problems during sectioning. It was very hard and caused section damage. There is always a possiblity that after you decalcify the shell, you can remove it very carefully to reach soft body parts. We had wonderful sections with thin shell intact - a total histological preparation. We decalcified in 10 to 15% formic acid after the snail was totally fixed. At 04:48 AM 9/8/2004, you wrote: >Torino 08 September 2004 >(ITALY) > > >Hi all, > >I am an amateur naturalist. >I like to study the histology of animal tissues by an optical microscope >in transmitted light. >I am interested to land (terrestrial) snails. >I know for relaxing the snail to use a solution of 50mM of MgCl2 by an >injection (2 ml.) into the foot. >Could someone give me some detailed information how to proceed to the >snail dissection? >Thank you. >With my Best Regards, > >Massimo 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 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 LINDABWILLIAMSON <@t> aol.com Wed Sep 8 11:25:07 2004 From: LINDABWILLIAMSON <@t> aol.com (LINDABWILLIAMSON@aol.com) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] registry tissue Message-ID: <148.33267272.2e708c63@aol.com> I need uterus 2 cm x 2 cm, kidney 1.5 cm in length, artery 0.5 outside diameter. TRYING TO GET THE PREFECT SECTION, I cut my blocks too thin! My practical is due October 1. Any help would be greatly appreciated. thanks. Linda Williamson-Houston Texas. From David.Edmondson <@t> christie-tr.nwest.nhs.uk Wed Sep 8 11:31:32 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Tissue dye Message-ID: The Davidson's Marking System PO Box 201405, Bloomington MN 55420 www.bradleyproducts.com is what we use, I think the yellow does not survive processing well but the others we are told are fine. David Christie Manchester UK -----Original Message----- From: Celebre Julia [mailto:celebrej@HHSC.CA] Sent: 08 September 2004 12:13 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Tissue dye Morning all. Can anyone tell me what they use to ink their tissues prior to processing? We need to find a second and third tissue dye marker for our Pathologist Assistant since our supplier has discontinued the ones we currently use. Any suggestions are greatly appreciated. Thanks Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca 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 ihc <@t> unipathllc.com Wed Sep 8 11:43:04 2004 From: ihc <@t> unipathllc.com (UniPath IHC) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Parvovirus/Thank you Message-ID: <000001c495c2$e9c03b60$4500a8c0@unipathllc.corp> Hello Everyone, Thank you so much to everyone who gave me guidance on the Mallory's PTAH for my practical. It worked the best when I did the Ann Preece treatment in Bouins followed by the Cherukian method. It's not perfect, but I gave it my best shot and I know all of the protocols inside and out, so I feel pretty good about it. I'm calling it Cherukian with a twist. Is anyone aware of a source for control slides for Parvovirus B19? We need positive tissue to work the antibody up. Thanks again, Brianna Jackson, BS, QIHC UniPath, LLC Denver, CO From jluis.palazon <@t> icman.csic.es Wed Sep 8 11:56:54 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Land snail dissection. Message-ID: <20040908165654.8F9A12A8021@perceval.uca.es> If the snail is small I recomend you to fix the whole snail and after fixation, decalcify it with 10 % EDTA. then process and include the whole snail. Hope this help El dia 08/09/2004 18:23 usted envio el siguiente mensaje: >Date: 8 de Septiembre de 2004 18:23:01 >From: "Smith, Allen" >Subject: RE: [Histonet] Land snail dissection. >To: gcallis@montana.edu, histonet@pathology.swmed.edu > > Many centuries ago, I forced a snail out of its shell by shredding a pack of > cigarettes into a pint of water and dropping the snail into it. > Borradaile's THE INVERTEBRATA has instructions for dissecting the European > garden snail Helix pomatia (pp. 604-610 in the 4th edition). The book is > out of print, but available used. > > 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 Gayle Callis > Sent: Wednesday, September 08, 2004 10:56 AM > To: andromeda_tm; Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Land snail dissection. > > > > Becaue of the hard calcified shell, dissection was still difficult. The > snail didn't always relax all the way out of the shell. I think we also > used another way to relax snail, but do not recall what was used (many > years ago!), as long as your method works, use it. > > Since we could never get a snail totally out of shell (they don't give it > up easily!), we fixed snails whole, then decalcified them with shells > intact, and processed them into paraffin. There is a "tooth" of some sort, > I do not recall what my snail experts called it, but it can create problems > during sectioning. It was very hard and caused section damage. There is > always a possiblity that after you decalcify the shell, you can remove it > very carefully to reach soft body parts. We had wonderful sections with > thin shell intact - a total histological preparation. > > We decalcified in 10 to 15% formic acid after the snail was totally fixed. > > At 04:48 AM 9/8/2004, you wrote: > >Torino 08 September 2004 > >(ITALY) > > > > > >Hi all, > > > >I am an amateur naturalist. > >I like to study the histology of animal tissues by an optical microscope > >in transmitted light. > >I am interested to land (terrestrial) snails. > >I know for relaxing the snail to use a solution of 50mM of MgCl2 by an > >injection (2 ml.) into the foot. > >Could someone give me some detailed information how to proceed to the > >snail dissection? > >Thank you. > >With my Best Regards, > > > >Massimo > > 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 > > > > 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) > > _______________________________________________ > 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 jluis.palazon <@t> icman.csic.es Wed Sep 8 11:57:44 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:24:00 2005 Subject: Re[Histonet] plastic in paraffin block Message-ID: <20040908165744.991E12A53A7@perceval.uca.es> Wen, I suppose that the plastic will disolve in the xilol so it would not be a problem El dia 08/09/2004 18:15 usted envio el siguiente mensaje: > ----- Original Message ----- > From: "wen eng" > To: > Sent: Tuesday, September 07, 2004 8:54 PM > Subject: [Histonet] plastic in paraffin block > > > > Hi, > > > > I got some skin tissue in formalin that covered with plastic film. If I > peel off plastic it will damaged the wound. Looks like I have to process > them with the plastic on. But I worried that the plastic will give me big > headache when I section them. > > > > Anybody has good idea? > > > > Thanks in advance! > > > > Wen > > > > > > > > --------------------------------- > > 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 > > > > > _______________________________________________ > 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 Bauer.Karen <@t> mayo.edu Wed Sep 8 12:09:47 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Tissue dye Message-ID: <8C6E05FA69571948B461F1327CBB893E0629F143@lmmail2.ad.lmmhs.org> I'm with David. We also use the Davidson Marking System and it works great for us. We have Blue, Green, Yellow, Black, Red and Orange. The orange tends to turn our solutions in the processor orange, so we don't like to use it very much. Only one Pathologist really uses it, since he likes the color and consistency of the dye. We all have our favorite color that we like to use and the variety is nice. The dye stays on the tissue nicely and they are very easy to use. Visit the website that David gave, it will show you what they have. Also, the caps with the little brushes on them are wonderful. Karen Bauer HT(ASCP) Histology Supervisor Department of Pathology Luther Hospital Eau Claire, WI -----Original Message----- From: Edmondson David (RBV) NHS Christie Tr [mailto:David.Edmondson@christie-tr.nwest.nhs.uk] Sent: Wednesday, September 08, 2004 11:32 AM To: 'Celebre Julia' Cc: Histonet (E-mail 2) Subject: RE: [Histonet] Tissue dye The Davidson's Marking System PO Box 201405, Bloomington MN 55420 www.bradleyproducts.com is what we use, I think the yellow does not survive processing well but the others we are told are fine. David Christie Manchester UK -----Original Message----- From: Celebre Julia [mailto:celebrej@HHSC.CA] Sent: 08 September 2004 12:13 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Tissue dye Morning all. Can anyone tell me what they use to ink their tissues prior to processing? We need to find a second and third tissue dye marker for our Pathologist Assistant since our supplier has discontinued the ones we currently use. Any suggestions are greatly appreciated. Thanks Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From pruegg <@t> ihctech.net Wed Sep 8 12:33:01 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Specificity of Masson's trichrome In-Reply-To: <005d01c49541$cf1ddc30$8a19fea9@walkercvzdlx4z> Message-ID: <002a01c495c9$e3f5d300$83020a0a@IHCTech> I find the Pentachrome stain is better than trichrome for lung samples. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Walker/Langdon Sent: Tuesday, September 07, 2004 6:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Specificity of Masson's trichrome Does anyone have details regarding the specificity of the collagen staining when tissues(especially lung) are stained with Masson's trichrome? Is it possible that fibronectin and/or alpha smooth muscle actin can also be stained? We are trying to reconcile some IHC and in vitro data. Thanks so much! Carrie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Wed Sep 8 12:39:46 2004 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] chromogen Message-ID: <002b01c495ca$d5a37bd0$83020a0a@IHCTech> What is a good substitute for DAB using HRP, my investigator wants prettier red or green colors for a poster, something like he gets with FITC, but I don't want to do flourescence? I know I could use AEC but I would prefer something more permanent. Didn't Kirkgaurd and Perry come out with a permanent red chromogen for HRP? Patsy From Jackie.O'Connor <@t> abbott.com Wed Sep 8 13:09:21 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Land snail dissection. Message-ID: Out of curiosity - is the shell made of calcium? I'm asking because I really don't know - not a trick question? Isn't a snail out of it's shell just a slug? (Now THAT is a joke.) Jackie O' Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics 847.938.4919 Fax 847.938.3266 Jose Luis Palazon Fernandez Sent by: histonet-bounces@lists.utsouthwestern.edu 09/08/2004 11:56 AM To: histonet@pathology.swmed.edu cc: Subject: Re: RE: [Histonet] Land snail dissection. If the snail is small I recomend you to fix the whole snail and after fixation, decalcify it with 10 % EDTA. then process and include the whole snail. Hope this help El dia 08/09/2004 18:23 usted envio el siguiente mensaje: >Date: 8 de Septiembre de 2004 18:23:01 >From: "Smith, Allen" >Subject: RE: [Histonet] Land snail dissection. >To: gcallis@montana.edu, histonet@pathology.swmed.edu > > Many centuries ago, I forced a snail out of its shell by shredding a pack of > cigarettes into a pint of water and dropping the snail into it. > Borradaile's THE INVERTEBRATA has instructions for dissecting the European > garden snail Helix pomatia (pp. 604-610 in the 4th edition). The book is > out of print, but available used. > > 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 Gayle Callis > Sent: Wednesday, September 08, 2004 10:56 AM > To: andromeda_tm; Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Land snail dissection. > > > > Becaue of the hard calcified shell, dissection was still difficult. The > snail didn't always relax all the way out of the shell. I think we also > used another way to relax snail, but do not recall what was used (many > years ago!), as long as your method works, use it. > > Since we could never get a snail totally out of shell (they don't give it > up easily!), we fixed snails whole, then decalcified them with shells > intact, and processed them into paraffin. There is a "tooth" of some sort, > I do not recall what my snail experts called it, but it can create problems > during sectioning. It was very hard and caused section damage. There is > always a possiblity that after you decalcify the shell, you can remove it > very carefully to reach soft body parts. We had wonderful sections with > thin shell intact - a total histological preparation. > > We decalcified in 10 to 15% formic acid after the snail was totally fixed. > > At 04:48 AM 9/8/2004, you wrote: > >Torino 08 September 2004 > >(ITALY) > > > > > >Hi all, > > > >I am an amateur naturalist. > >I like to study the histology of animal tissues by an optical microscope > >in transmitted light. > >I am interested to land (terrestrial) snails. > >I know for relaxing the snail to use a solution of 50mM of MgCl2 by an > >injection (2 ml.) into the foot. > >Could someone give me some detailed information how to proceed to the > >snail dissection? > >Thank you. > >With my Best Regards, > > > >Massimo > > 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 > > > > 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) > > _______________________________________________ > 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 DayDawning <@t> wideopenwest.com Wed Sep 8 13:13:16 2004 From: DayDawning <@t> wideopenwest.com (Dawn Truscott) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] snails References: <200409081720.i88HK3k23416@smtp-3.wideopenwest.com> Message-ID: <000501c495cf$832f4aa0$6401a8c0@wowway.com> I'm thinking Garlic and butter..... > > > Hi all, > > I am an amateur naturalist. > I like to study the histology of animal tissues by an optical microscope in transmitted light. > I am interested to land (terrestrial) snails. > I know for relaxing the snail to use a solution of 50mM of MgCl2 by an injection (2 ml.) into the foot. > Could someone give me some detailed information how to proceed to the snail dissection? > Thank you. > With my Best Regards, > > Massimo > From dlcowie <@t> prodigy.net Wed Sep 8 13:14:30 2004 From: dlcowie <@t> prodigy.net (Dawn Cowie) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] CLIA regs Message-ID: <20040908181430.27137.qmail@web81010.mail.yahoo.com> hi all, Does anyone know if there is any CLIA reg that covers maintaining records of slides and or blocks that are sent out and then returned from outside facilities? I know per CAP that you must keep blocks and slides 10 years but this doesn't really cover this aspect. Any feedback will be appreciated. Thanks in advance, Dawn Cowie, HT Histology Supv Pensacola Path From Joan.Sempf <@t> med.va.gov Wed Sep 8 13:18:56 2004 From: Joan.Sempf <@t> med.va.gov (Sempf, Joan M.) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Sorvall TC-2 Tissue Sectoner Message-ID: I have just found a Sorvall TC-2 Tissue Sectioner and have no instruction manual to go with it. Our graduate student would like to do 50 micron sections for pre-embedding immunogold. Is there any one who could help me out with its operation? Thanks Joan Sempf William S. Middleton Mem. VAMC Department of Pathology 2500 Overlook Terrace Madison, WI 53705 From CMCCOLLOUGH <@t> dnr.state.md.us Wed Sep 8 13:46:40 2004 From: CMCCOLLOUGH <@t> dnr.state.md.us (McCollough, Carol) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Land snail dissection & other shellfish Message-ID: You folks speaking of garlic & butter... Over 1300 live oysters pass through my histology lab each fall, plus over 300 New England steamers (clams) - that's 108 dozen oysters on the half-shell and 27 dozen steamers (about 18 buckets) - and we can't eat a one of 'em :-( I can't imagine how the shrimp pathology folks feel........ To add insult to injury, we use Davidson's fixative, and the acetic acid smell makes my mouth water and stomach growl just before we begin shucking. Regards - Carol ********************** Carol B. McCollough, HT/HTL(ASCP) Diagnostics & Histology Laboratory Manager Oyster Disease Research Project Fisheries Service Maryland Department of Natural Resources Cooperative Oxford Laboratory 904 S. Morris Street Oxford, Maryland 21654 410-226-5193 From gcallis <@t> montana.edu Wed Sep 8 14:00:07 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Shredded cigarettes for Land snail dissection. In-Reply-To: <4C051EAE581BB646BF53A749A73FBA2D1F3BDF@exchsrv01.barrynet. barry.edu> References: <4C051EAE581BB646BF53A749A73FBA2D1F3BDF@exchsrv01.barrynet.barry.edu> Message-ID: <6.0.0.22.1.20040908125416.01b0ea18@gemini.msu.montana.edu> At the risk of too much humor, would chewing tobacco (not used, please!) do the job instead of shredding cigarettes. A scientific use for SKOL!! At 10:23 AM 9/8/2004, you wrote: >Many centuries ago, I forced a snail out of its shell by shredding a pack of >cigarettes into a pint of water and dropping the snail into it. >Borradaile's THE INVERTEBRATA has instructions for dissecting the European >garden snail Helix pomatia (pp. 604-610 in the 4th edition). The book is >out of print, but available used. > >Allen A. Smith, Ph.D. >Professor of Anatomy >Barry University >School of Graduate Medical Sciences > Podiatric Medicine and Surgery >Miami Shores, Florida Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From rgrow <@t> bmnet.com Wed Sep 8 14:01:14 2004 From: rgrow <@t> bmnet.com (rgrow@bmnet.com) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Re:Tissue Dye In-Reply-To: <200409081720.i88HKqoB003590@dns1.bmnet.com> Message-ID: Julia, For routine FFPE sections give Cancer Diagnostics, Inc. a call at 1-877-846-5393 or visit their website: www.cancerdiagnostics.com. They have a good selection to choose from. We use all their colors (green, black, blue, red, yellow, orange) and are testing the new purple at this time. And yes, we have used all colors at once to ink complicated specimens. They look very sharp under the scope. A quick dip in bouins solution before sectioning and processing helps "set" the dye, a step this lab has used for a long time. I'm not sure it's necessary now. They are a small company that offers a good product. Good luck, Renee Grow, BA., HT (ASCP) rgrow@bmnet.com Histology Supervisor Blount Memorial Hospital 907 E. Lamar Alexander Pkwy. Maryville, TN 37804-5016 (865) 977-4744 (865) 977-5766 Fax -----Original Message----- From: Celebre Julia [mailto:celebrej@HHSC.CA] Sent: 08 September 2004 12:13 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Tissue dye Morning all. Can anyone tell me what they use to ink their tissues prior to processing? We need to find a second and third tissue dye marker for our Pathologist Assistant since our supplier has discontinued the ones we currently use. Any suggestions are greatly appreciated. Thanks Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca From gcallis <@t> montana.edu Wed Sep 8 14:29:26 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] chromogen In-Reply-To: <002b01c495ca$d5a37bd0$83020a0a@IHCTech> References: <002b01c495ca$d5a37bd0$83020a0a@IHCTech> Message-ID: <6.0.0.22.1.20040908132243.01b0dc00@gemini.msu.montana.edu> DAKO has Permanent Red for alk phos IHC. Chris van der Loos indicated DAKO product was very sensitive, and it also fluoresces. Vector red is AP and also fluoresces. Have not tried Vector Red to compare with primary antibody concentrations. These reds are more on dark pink red side, rather than orangish red of AEC, or Brick red of NOVA Red. NOVA Red from Vector, but it is a brick red!! Darker than AEC. You can mount AEC from water, using Crystal mount, let this dry and then add a permanent mounting media. At 11:39 AM 9/8/2004, you wrote: >What is a good substitute for DAB using HRP, my investigator wants >prettier red or green colors for a poster, something like he gets with >FITC, but I don't want to do flourescence? I know I could use AEC but I >would prefer something more permanent. Didn't Kirkgaurd and Perry come >out with a permanent red chromogen for HRP? >Patsy >_______________________________________________ >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 haldana <@t> unimoron.edu.ar Wed Sep 8 14:29:22 2004 From: haldana <@t> unimoron.edu.ar (Hernan Aldana) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Stem cells quantification with cd in blood In-Reply-To: <200409081410265.SM01472@swlx162.swmed.edu> Message-ID: <001301c495da$258eef30$b58d72c8@b3w6zzmtb6juvbs> I need to make Stem cell quantification in blood with CD43 or other cell maker. I need to use flow cytometry, does anybody have some experience with it methodology? Or other? 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 Jackie.O'Connor <@t> abbott.com Wed Sep 8 14:31:59 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Land snail dissection & other shellfish Message-ID: I've spent my career rinsing out colons, slicing through eyes with a scalpel, looking for the depth of gangrene in half a foot - but the thought of eating a snail literally makes me sick. I've watched "Fear Factor" when the contestants have to eat pig rectum and thought "what's the big deal" - but when they had to bite into a live cave cricket - I had to leave the room. As far as I'm concerned, snails are in the same family. I almost had to go to the ER once when, living in California, my bare foot stepped on a slug that had crawled under my back door into the laundry room. But plop a whole digestive tract in front of me to clean - no problem. Creepy. "McCollough, Carol" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/08/2004 01:46 PM To: cc: Subject: RE: [Histonet] Land snail dissection & other shellfish You folks speaking of garlic & butter... Over 1300 live oysters pass through my histology lab each fall, plus over 300 New England steamers (clams) - that's 108 dozen oysters on the half-shell and 27 dozen steamers (about 18 buckets) - and we can't eat a one of 'em :-( I can't imagine how the shrimp pathology folks feel........ To add insult to injury, we use Davidson's fixative, and the acetic acid smell makes my mouth water and stomach growl just before we begin shucking. Regards - Carol ********************** Carol B. McCollough, HT/HTL(ASCP) Diagnostics & Histology Laboratory Manager Oyster Disease Research Project Fisheries Service Maryland Department of Natural Resources Cooperative Oxford Laboratory 904 S. Morris Street Oxford, Maryland 21654 410-226-5193 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Wed Sep 8 17:36:28 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Shredded cigarettes for Land snail dissection. In-Reply-To: <6.0.0.22.1.20040908125416.01b0ea18@gemini.msu.montana.edu> References: <4C051EAE581BB646BF53A749A73FBA2D1F3BDF@exchsrv01.barrynet.barry.edu> <6.0.0.22.1.20040908125416.01b0ea18@gemini.msu.montana.edu> Message-ID: <413F896C.7070500@umdnj.edu> I think so, I think it is the nicotine that is the narcotic here. Geoff Gayle Callis wrote: > At the risk of too much humor, would chewing tobacco (not used, > please!) do the job instead of shredding cigarettes. A scientific use > for SKOL!! > > At 10:23 AM 9/8/2004, you wrote: > >> Many centuries ago, I forced a snail out of its shell by shredding a >> pack of >> cigarettes into a pint of water and dropping the snail into it. >> Borradaile's THE INVERTEBRATA has instructions for dissecting the >> European >> garden snail Helix pomatia (pp. 604-610 in the 4th edition). The >> book is >> out of print, but available used. >> >> Allen A. Smith, Ph.D. >> Professor of Anatomy >> Barry University >> School of Graduate Medical Sciences >> Podiatric Medicine and Surgery >> Miami Shores, Florida > > > 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 > -- -- ********************************************** 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 DDittus787 <@t> aol.com Wed Sep 8 15:30:50 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] CLIA regs Message-ID: <229B7C27.217A5085.0A1F969F@aol.com> Clia bows to the most stringent rule committee , so if the cap rules are more stringent than state or other , clia will go with cap. Dana From gcallis <@t> montana.edu Wed Sep 8 15:33:24 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] snails and slugs In-Reply-To: References: Message-ID: <6.0.0.22.1.20040908141939.01b0d7a0@gemini.msu.montana.edu> The shell is calcified but not pure calcium, so there is a protein type matrix there, not sure what matrix consists of?? A relaxed snail only comes partially out of its shell, and never really reaches true "slug" stage!! Snails are tenacious and only like to "hang out" rather than "fall out" completely from shell. Hmmm for quick return into protective calcified "home"? By the way, I have an un-opened jar "Slug" preserves from Seattle - where 10 billion slugs can't be wrong and love to live! And you haven't lived until you have been slimed by a slug! Firsthand knowledge and a hilarious story. Enough of this slimey conversation! Gayle Callis At 12:09 PM 9/8/2004, you wrote: >Out of curiosity - is the shell made of calcium? I'm asking because I >really don't know - not a trick question? Isn't a snail out of it's shell >just a slug? >(Now THAT is a joke.) > > >Jackie O' > > >Jacqueline M. O'Connor HT(ASCP) >Abbott Laboratories >Global Pharmaceutical Research and Development >Discovery Chemotherapeutics >847.938.4919 >Fax 847.938.3266 > From peoshel <@t> wisc.edu Wed Sep 8 15:38:30 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Land snail dissection. Message-ID: Yep, CaCO3. Also yes. But, why bother with decalification? Just play crab and crack the shell open. The snail can be removed alive, if unhappy. I'd put it in MgCl2 first, then open the shell and remove the snail, cool it to further relax and anesthetize it, inject fixative into the mantle cavity (and possibly the hemocoel), then immersion-fix it. For sectioning, I'd dissect the snail into smaller pieces to insure proper fixation and infiltration paraffin -- they have a very tough, muscular foot, and the mantle can be as well. The radula that Gayle referred to earlier is mostly keratin, but many snails deposit CaCO3 or other minerals (including iron, if I remember right) in the tips of the radular teeth -- either way, it will cause grief when paraffin sectioning. It'd be better to carefully dissect away the radula and mount it whole -- whole mounts of radulae are used in molluc taxonomy anyway. If you do want to section the radula, you will need to plastic embed it. Phil >Out of curiosity - is the shell made of calcium? I'm asking because I >really don't know - not a trick question? Isn't a snail out of it's shell >just a slug? >(Now THAT is a joke.) > > >Jackie O' > > >Jacqueline M. O'Connor HT(ASCP) >Abbott Laboratories >Global Pharmaceutical Research and Development >Discovery Chemotherapeutics >847.938.4919 >Fax 847.938.3266 > > > > >Jose Luis Palazon Fernandez >Sent by: histonet-bounces@lists.utsouthwestern.edu >09/08/2004 11:56 AM > > > To: histonet@pathology.swmed.edu > cc: > Subject: Re: RE: [Histonet] Land snail dissection. > > >If the snail is small I recomend you to fix the whole snail and after >fixation, decalcify it with 10 % EDTA. then process and include the whole >snail. Hope this help > > > > > >El dia 08/09/2004 18:23 usted envio el siguiente mensaje: > >>Date: 8 de Septiembre de 2004 18:23:01 > >>From: "Smith, Allen" > >>Subject: RE: [Histonet] Land snail dissection. > >>To: gcallis@montana.edu, histonet@pathology.swmed.edu > >> > >> Many centuries ago, I forced a snail out of its shell by shredding a >pack of > >> cigarettes into a pint of water and dropping the snail into it. > >> Borradaile's THE INVERTEBRATA has instructions for dissecting the >European > >> garden snail Helix pomatia (pp. 604-610 in the 4th edition). The book >is > >> out of print, but available used. > >> > >> 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 Gayle >Callis > >> Sent: Wednesday, September 08, 2004 10:56 AM > >> To: andromeda_tm; Histonet@lists.utsouthwestern.edu > >> Subject: [Histonet] Land snail dissection. > >> > >> > >> > >> Becaue of the hard calcified shell, dissection was still difficult. The > > >> snail didn't always relax all the way out of the shell. I think we also > > >> used another way to relax snail, but do not recall what was used (many > >> years ago!), as long as your method works, use it. > >> > >> Since we could never get a snail totally out of shell (they don't give >it > >> up easily!), we fixed snails whole, then decalcified them with shells > >> intact, and processed them into paraffin. There is a "tooth" of some >sort, > >> I do not recall what my snail experts called it, but it can create >problems > >> during sectioning. It was very hard and caused section damage. There >is > >> always a possiblity that after you decalcify the shell, you can remove >it > >> very carefully to reach soft body parts. We had wonderful sections with > > >> thin shell intact - a total histological preparation. > >> > >> We decalcified in 10 to 15% formic acid after the snail was totally >fixed. > >> > >> At 04:48 AM 9/8/2004, you wrote: > >> >Torino 08 September 2004 > >> >(ITALY) > >> > > >> > > >> >Hi all, > >> > > >> >I am an amateur naturalist. > >> >I like to study the histology of animal tissues by an optical >microscope > >> >in transmitted light. > >> >I am interested to land (terrestrial) snails. > >> >I know for relaxing the snail to use a solution of 50mM of MgCl2 by an > >> >injection (2 ml.) into the foot. > >> >Could someone give me some detailed information how to proceed to the > >> >snail dissection? > >> >Thank you. > >> >With my Best Regards, > >> > > >> >Massimo > >> > >> 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 > >> > >> > >> > >> 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) > >> > >> _______________________________________________ > >> 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 > > >_______________________________________________ >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 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From peoshel <@t> wisc.edu Wed Sep 8 15:41:36 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] snails and slugs In-Reply-To: <6.0.0.22.1.20040908141939.01b0d7a0@gemini.msu.montana.edu> References: <6.0.0.22.1.20040908141939.01b0d7a0@gemini.msu.montana.edu> Message-ID: Collagen, mostly, standard connective tissue. I'd have to get out the gastropod volume of "Microscopic Anatomy of Invertebrates" to be sure. Mollusc shell microarchitecture is pretty neat, really. Yeah, I'm surprised a snail could be gotten out of its shell by any means -- slugs may be shell-less snails, but snails without shells are dead, they just may not know it yet. Phil >The shell is calcified but not pure calcium, so there is a protein >type matrix there, not sure what matrix consists of?? A relaxed >snail only comes partially out of its shell, and never really >reaches true "slug" stage!! Snails are tenacious and only like to >"hang out" rather than "fall out" completely from shell. Hmmm for >quick return into protective calcified "home"? > >By the way, I have an un-opened jar "Slug" preserves from Seattle - >where 10 billion slugs can't be wrong and love to live! And you >haven't lived until you have been slimed by a slug! Firsthand >knowledge and a hilarious story. > >Enough of this slimey conversation! > >Gayle Callis > > > >At 12:09 PM 9/8/2004, you wrote: >>Out of curiosity - is the shell made of calcium? I'm asking because I >>really don't know - not a trick question? Isn't a snail out of it's shell >>just a slug? >>(Now THAT is a joke.) >> >> >>Jackie O' >> >> >>Jacqueline M. O'Connor HT(ASCP) >>Abbott Laboratories >>Global Pharmaceutical Research and Development >>Discovery Chemotherapeutics >>847.938.4919 >>Fax 847.938.3266 >> > > >_______________________________________________ >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 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From JNocito <@t> Pathreflab.com Wed Sep 8 16:00:36 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] CLIA regs In-Reply-To: <20040908181430.27137.qmail@web81010.mail.yahoo.com> Message-ID: Dawn, when I had my CLIA inspection a couple of years ago, the only thing that was brought up was how long we keep consult blocks and slides before we return them to the submitting facility. Our standard answer is at least 2 weeks after the case is signed out. 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 Dawn Cowie Sent: Wednesday, September 08, 2004 1:15 PM To: histonet Subject: [Histonet] CLIA regs hi all, Does anyone know if there is any CLIA reg that covers maintaining records of slides and or blocks that are sent out and then returned from outside facilities? I know per CAP that you must keep blocks and slides 10 years but this doesn't really cover this aspect. Any feedback will be appreciated. Thanks in advance, Dawn Cowie, HT Histology Supv Pensacola Path _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From caroline.stott <@t> anatomy.otago.ac.nz Wed Sep 8 17:00:26 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Resin embedding-bone matrix Message-ID: <5.2.1.1.0.20040909095304.02a40e30@anatomy.otago.ac.nz> Hi all, I don't think this will work but.... I have been asked to resin embed some bone that was exposed to treatments of high temps and pressures etc. Consequently I have been left only with the in-organic matrix. I cant decalcify it, or embed in paraffin because there will be nothing left, and I don't think it will cut even if I manage to get it into resin. One of the samples is just powder!!! (Apparently they are looking at a particular protein?) However the question is, should I make my resin mixture harder than normal to accommodate the hard nature of the tissue? Has anyone tried anything similar??? Or do you have another medium I can embed into? Help and many thanks Caroline Caroline Stott Histology Service Unit University of Otago PO Box 913 Dunedin, New Zealand Ph (03) 479 7152 Fax (03) 479 7136 From weneng2004 <@t> yahoo.com Wed Sep 8 17:27:18 2004 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Thank you!/Plastic in paraffin block. Message-ID: <20040908222718.32301.qmail@web53409.mail.yahoo.com> Thank you to all who gave me advices on dealing with plastic in paraffin block! The film is called Tegaderm, the product from 3M. I leave the sample in formalin for a longer time. at the same time, as you suggested, I cut a small piece into Xylene. But it is still there the next morning :(. So xylene can not disolve this material. I didn't test it in xylol. When I came back to my samples, I found the film is much easier to be peeled off now after a longer time fixation. Thank you again! Wen --------------------------------- Do you Yahoo!? Shop for Back-to-School deals on Yahoo! Shopping. From JPR <@t> Stowers-Institute.org Wed Sep 8 17:39:57 2004 From: JPR <@t> Stowers-Institute.org (Rey, Jean-Philippe) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] How obtain a Methyl Green counterstain after a Citrate pretreatment? Message-ID: <1117439498-141377934@pathology.swmed.edu> Hi everyone, I try to counterstain some immunostained sections with Methyl green. The one which was pretreated with Citrate buffer at 121degre Celcius, can't been stained with Methyl Green. Does some one have a clue why? And to be able to obtain some Methyl Green after a slide undergo a Citrate pretreatment? I will appreciate your help. Have a nice day Jean-Philippe Rey Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 (816) 926-4305 From gcallis <@t> montana.edu Wed Sep 8 17:58:10 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Resin embedding-bone matrix In-Reply-To: <5.2.1.1.0.20040909095304.02a40e30@anatomy.otago.ac.nz> References: <5.2.1.1.0.20040909095304.02a40e30@anatomy.otago.ac.nz> Message-ID: <6.0.0.22.1.20040908165615.01ae95f0@gemini.msu.montana.edu> You did not say what plastic you are using. Methyl methacrylate is considerably harder than glycol methacrylate. At 04:00 PM 9/8/2004, you wrote: >Hi all, >I don't think this will work but.... I have been asked to resin embed some >bone that was exposed to treatments of high temps and pressures >etc. Consequently I have been left only with the in-organic matrix. I >cant decalcify it, or embed in paraffin because there will be nothing >left, and I don't think it will cut even if I manage to get it into >resin. One of the samples is just powder!!! (Apparently they are looking >at a particular protein?) >However the question is, should I make my resin mixture harder than normal >to accommodate the hard nature of the tissue? Has anyone tried anything >similar??? Or do you have another medium I can embed into? >Help and many thanks >Caroline > >Caroline Stott > >Histology Service Unit >University of Otago >PO Box 913 >Dunedin, New Zealand >Ph (03) 479 7152 >Fax (03) 479 7136 > >_______________________________________________ >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 caroline.stott <@t> anatomy.otago.ac.nz Wed Sep 8 18:55:35 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] Resin embedding-bone matrix Message-ID: <5.2.1.1.0.20040909114531.029dc580@anatomy.otago.ac.nz> Sorry, we will probably try with Glycol methacrylate because I've used it many times and haven't tried Methyl Methacrylate. But if someone thinks MMA will be more beneficial? Or something else? I was going to try a 90 (2-hydroxymethacrylate) to 10 (2-butoxyethanol) mix, instead of the normal 88-12 mix. I am thinking of putting the bone straight into the promoted mix and skipping the infiltrating. Just placing it all in the freezer overnight, then into the fridge the next day to polymerize. Anyone have any ideas? Thanks Caroline Caroline Stott Histology Service Unit University of Otago PO Box 913 Dunedin, New Zealand Ph (03) 479 7152 Fax (03) 479 7136 From info <@t> ihcworld.com Wed Sep 8 19:01:30 2004 From: info <@t> ihcworld.com (IHC World) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] How obtain a Methyl Green counterstain after a Citratepretreatment? In-Reply-To: <1117439498-141377934@pathology.swmed.edu> Message-ID: Methyl green is excellent for IHC counterstain when nuclear markers is used. It will not overstain nucleus. However, some tissues have low affinity to methyl green. In this case you will get no or weak nuclear staining. Use the following protocol may help you solve problems if you haven't done so. http://www.ihcworld.com/counterstain_solutions.htm Good luck. Richard -----Original Message----- From: Rey, Jean-Philippe [mailto:JPR@Stowers-Institute.org] Sent: Wednesday, September 08, 2004 5:40 PM To: Histonet request (E-mail) Subject: [Histonet] How obtain a Methyl Green counterstain after a Citratepretreatment? Hi everyone, I try to counterstain some immunostained sections with Methyl green. The one which was pretreated with Citrate buffer at 121degre Celcius, can't been stained with Methyl Green. Does some one have a clue why? And to be able to obtain some Methyl Green after a slide undergo a Citrate pretreatment? I will appreciate your help. Have a nice day Jean-Philippe Rey Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 (816) 926-4305 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JAnorthernexp <@t> cs.com Wed Sep 8 19:06:42 2004 From: JAnorthernexp <@t> cs.com (JAnorthernexp@cs.com) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] synaptophysin Message-ID: <1cb.2a886bef.2e70f892@cs.com> anyone using a ventana stainer and antibodies have a retrieval method using a decloker? Ours stains very light. Antibody is on for 32 mins we did have it set at 16mins. But increasing the time did not prove to make much difference. Any other ideas?? The tissue is formalin fixed and paraffin embedded. Thanks Annette From jnocito <@t> satx.rr.com Wed Sep 8 21:01:56 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] synaptophysin References: <1cb.2a886bef.2e70f892@cs.com> Message-ID: <003c01c49610$fc2f2dd0$4661ce44@yourxhtr8hvc4p> Annette, did you try increasing the time in cell conditioner? Have you tried to amplify it? Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: To: Sent: Wednesday, September 08, 2004 7:06 PM Subject: [Histonet] synaptophysin > anyone using a ventana stainer and antibodies have a retrieval method using a > decloker? Ours stains very light. Antibody is on for 32 mins we did have it > set at 16mins. But increasing the time did not prove to make much > difference. Any other ideas?? The tissue is formalin fixed and paraffin embedded. > Thanks > > Annette > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From karrie.langdon <@t> sympatico.ca Wed Sep 8 22:48:17 2004 From: karrie.langdon <@t> sympatico.ca (Walker/Langdon) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] specificity of Masson's trichrome Message-ID: <000d01c4961f$db08dc00$8a19fea9@walkercvzdlx4z> Hi Histonetters, To clarify my question of the other day: is it true that Masson's trichrome could potentially stain fibronectin and/or smooth muscle actin, especially if it is being overexpressed? Is there a histochemical stain for fibronectin? We have already successfully done IHC,but are trying to reconcile data obtained with a few different approaches. We are mostly working in lung. Thanks to those who have already provided ideas! Carrie Langdon Dept. of Pathology McMaster University Hamilton, Ontario Canada From int09018 <@t> alphahunt.com Wed Sep 8 23:18:46 2004 From: int09018 <@t> alphahunt.com (HCS) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] needed: used Fisher 166A or 166MP tissue processor Message-ID: <000701c49624$1a104740$6501a8c0@hp> Does anyone have an extra Fisher 166MP or 166A. Mine is beyond repair and would like to find a working unit. Please email directly. Hope the sales reps do not call, the budget does not allow for new units at this time. LeRoy Brown HT(ASCP) HTL HCS Everson, WA 98247 1-360-966-7300 From balajimr <@t> drreddys.com Thu Sep 9 01:34:53 2004 From: balajimr <@t> drreddys.com (balajimr@drreddys.com) Date: Fri Sep 16 15:24:00 2005 Subject: [Histonet] PCNA - URINARY BLADDER Message-ID: Dear Histonetters, It's nice to be back with Histosearch after a short break. Well, I am facing problem with PCN staining of urinary bladder of rat. We are using Zymed PCNA staining kit for the same. I am getting non specific staining where even the nucleus of the muscle layers are also staining. We are retrieving the antigen by heating the sections in Citrate buffer. But we are not having this problem in the intestines of same control animals. Does anybody have experience of PCNA staining with U. bladder so that they can advise me about this problem. Ofcourse we have increased the blocking time with serum upto 30 minutes unsuccessfully. Thanks in advance. Dr. M.R. Balaji Dept. Pre clinical safety evaluation, Discovery research, Dr. Reddys Laboratories ltd. Bollaram Road, Miyapur, Hyderabad, 500 049 Andhra Pradseh, INDIA Email - balajimr@drreddys.com Tel: 040- 23045439 - Ext.432. From rentonlf <@t> bru.wits.ac.za Thu Sep 9 02:05:13 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Re:Splendore-Hoeppli phenomenon Message-ID: <1094713513.8dd9db60rentonlf@bru.wits.ac.za> Now, if we could only use that in the lyrics of a song along the lines of: "lung.... has has Hoeppli-splendore thing" cheers -----Original Message----- From: "Marshall Terry Dr, Consultant Histopathologist" To: "Angela Bitting" , Date: Wed, 8 Sep 2004 14:19:44 +0100 Subject: RE: [Histonet] Modified acid fast stain for actinomyces No modification is needed. Neither does it stain the Actinomyces. Rather, the club shaped ends of the filaments, being an antigen-antibody complex - known as Splendore-Hoeppli phenomenon - are stained red. They are not necessarily present. 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: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: 08 September 2004 13:53 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Modified acid fast stain for actinomyces Anyone have a recipe for this stain? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From rentonlf <@t> bru.wits.ac.za Thu Sep 9 02:07:28 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Shredded cigarettes OT Message-ID: <1094713648.8dd9db60rentonlf@bru.wits.ac.za> Tobacco powder is still widely used by gardeners as a "natural/organic" insecticide -----Original Message----- From: Geoff McAuliffe To: Gayle Callis Date: Wed, 08 Sep 2004 15:36:28 -0700 Subject: Re: [Histonet] Shredded cigarettes for Land snail dissection. I think so, I think it is the nicotine that is the narcotic here. Geoff Gayle Callis wrote: > At the risk of too much humor, would chewing tobacco (not used, > please!) do the job instead of shredding cigarettes. A scientific use > for SKOL!! > > At 10:23 AM 9/8/2004, you wrote: > >> Many centuries ago, I forced a snail out of its shell by shredding a >> pack of >> cigarettes into a pint of water and dropping the snail into it. >> Borradaile's THE INVERTEBRATA has instructions for dissecting the >> European >> garden snail Helix pomatia (pp. 604-610 in the 4th edition). The >> book is >> out of print, but available used. >> >> Allen A. Smith, Ph.D. >> Professor of Anatomy >> Barry University >> School of Graduate Medical Sciences >> Podiatric Medicine and Surgery >> Miami Shores, Florida > > > 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 > -- -- ********************************************** 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 Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From carl.hobbs <@t> kcl.ac.uk Thu Sep 9 02:48:06 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] p75 and Neuro D Message-ID: <001701c49641$5805c3f0$e8345c9f@Carlos> Be most grateful if anyone out there can recommend antibodies against those two proteins, that work in mouse pwx sections. I have got Chemicon's p75 ( MAB 390). The p75 is the low affinity neurotrophin receptor. I have searched but I obviously am reluctant to buy unless confirmed that they work in pwax. Thank you From andromeda_tm <@t> libero.it Thu Sep 9 05:57:20 2004 From: andromeda_tm <@t> libero.it (Massimo) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] =?iso-8859-1?q?How_to_relax_a_land_snail=85definitive?= =?iso-8859-1?q?ly=2E?= Message-ID: Torino 09 September 2004 (ITALY) I would like to thank for your answers at my question about snail. I also think to do something pleasant and to raise the scientific contents of someone answers by sending them an Italian recipe on snails with garlic. Sorry in Italy we use the olive oil. Butter is very dangerous for the heart, the arteries ( just like the cigarettes smoke) and it also gets fat. I am afraid but the recipe is in Italian. But I?m sure if you enter a good Italian restaurant they can cook it. But remember do not forget to bring with you the snails! With my Best Regard Massimo Tosi Graduate Chemical Engineering PISA University ITALY LUMACHE CON L'AGLIO (SNAILS WITH GARLIC) INGREDIENTI PER 4 PERSONE 1Kg di lumache gi? spurgate (appena pulite vi consiglio di metterle per pi? di 2 ore al sole in un contenitore con abbondante sale sui bordi, le lumache infatti attirate dal sole e dal sale escono tutte fuori) 4 spicchi di aglio (GARLIC) 1 mazzetto di prezzemolo olio extravergine di oliva (OLIVE OIL) sale pepe Lessate le lumache per 15 minuti, schiumando con un mestolo forato ("schiumarola" appunto). A fine cottura, sciacquatele, rimettetele in tegame con acqua pulita leggermente salata e cuocete per altri 15 minuti. A questo punto, sgocciolatele (conservate per? un po' d'acqua di cottura) e trasferitele in una casseruola, dove avete fatto soffriggere per poco tempo l'aglio, il prezzemolo e il pepe, cuocete per pochi minuti e aggiustate di sale, irrorate con l'acqua di cottura e servite in una zuppiera con del prezzemolo fresco. E adesso inizia lo spasso.... buon appetito From AFeatherstone <@t> KaleidaHealth.Org Thu Sep 9 06:14:17 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] synaptophysin Message-ID: We antigen retrieve our synaptophysin in Citrate Buffer ph6.0 in a steamer. We set the steamer with the citrate in it for 70 minutes, at 20 minutes left we add the slides. After it steams for the final 20 minutes we let it cool for 20 minutes. We add amplifier and blocker too. Is your primary dilution correct? Annette Featherstone HT/MLT Kaleida Health. -----Original Message----- From: Joe Nocito [mailto:jnocito@satx.rr.com] Sent: Wednesday, September 08, 2004 22:02 To: JAnorthernexp@cs.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] synaptophysin Annette, did you try increasing the time in cell conditioner? Have you tried to amplify it? Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: To: Sent: Wednesday, September 08, 2004 7:06 PM Subject: [Histonet] synaptophysin > anyone using a ventana stainer and antibodies have a retrieval method using a > decloker? Ours stains very light. Antibody is on for 32 mins we did have it > set at 16mins. But increasing the time did not prove to make much > difference. Any other ideas?? The tissue is formalin fixed and paraffin embedded. > Thanks > > Annette > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From dobbin <@t> upei.ca Thu Sep 9 07:22:07 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Land snail dissection. In-Reply-To: Message-ID: <414020BF.22473.1CC595@localhost> Dear Phil (and others), All this talk of ripping open snail shells, injecting with MgCl2 and heaven forbid, fixing alive! Does anyone else out there have to work within the constraints of an Animal Care Committee? Some of you might be interested to hear that in a an academic setting, all procedures carried out on research animals (be it crustacean, fish, avian or mammalian) have to be approved under guidelines for ethical and humane use and care of animals for research. For instance, while I have no objection to boiling my lobsters alive at home, at work, lobsters must me properly anesthetized prior to humane euthanasia! It's a strange world we work in by times! Cheers! Greg Date sent: Wed, 08 Sep 2004 15:38:30 -0500 From: Philip Oshel To: Histonet@Pathology.swmed.edu Copies to: Subject: RE: [Histonet] Land snail dissection. > Yep, CaCO3. > Also yes. > But, why bother with decalification? Just play crab and crack the > shell open. The snail can be removed alive, if unhappy. I'd put it in > MgCl2 first, then open the shell and remove the snail, cool it to > further relax and anesthetize it, inject fixative into the mantle > cavity (and possibly the hemocoel), then immersion-fix it. For > sectioning, I'd dissect the snail into smaller pieces to insure > proper fixation and infiltration paraffin -- they have a very tough, > muscular foot, and the mantle can be as well. > The radula that Gayle referred to earlier is mostly keratin, but many > snails deposit CaCO3 or other minerals (including iron, if I remember > right) in the tips of the radular teeth -- either way, it will cause > grief when paraffin sectioning. It'd be better to carefully dissect > away the radula and mount it whole -- whole mounts of radulae are > used in molluc taxonomy anyway. If you do want to section the radula, > you will need to plastic embed it. > > Phil > > >Out of curiosity - is the shell made of calcium? I'm asking because I > >really don't know - not a trick question? Isn't a snail out of it's shell > >just a slug? > >(Now THAT is a joke.) > > > > > >Jackie O' > > > > > >Jacqueline M. O'Connor HT(ASCP) > >Abbott Laboratories > >Global Pharmaceutical Research and Development > >Discovery Chemotherapeutics > >847.938.4919 > >Fax 847.938.3266 > > > > > > > > > >Jose Luis Palazon Fernandez > >Sent by: histonet-bounces@lists.utsouthwestern.edu > >09/08/2004 11:56 AM > > > > > > To: histonet@pathology.swmed.edu > > cc: > > Subject: Re: RE: [Histonet] Land snail dissection. > > > > > >If the snail is small I recomend you to fix the whole snail and after > >fixation, decalcify it with 10 % EDTA. then process and include the whole > >snail. Hope this help > > > > > > > > > > > >El dia 08/09/2004 18:23 usted envio el siguiente mensaje: > > > >>Date: 8 de Septiembre de 2004 18:23:01 > > > >>From: "Smith, Allen" > > > >>Subject: RE: [Histonet] Land snail dissection. > > > >>To: gcallis@montana.edu, histonet@pathology.swmed.edu > > > >> > > > >> Many centuries ago, I forced a snail out of its shell by shredding a > >pack of > > > >> cigarettes into a pint of water and dropping the snail into it. > > > >> Borradaile's THE INVERTEBRATA has instructions for dissecting the > >European > > > >> garden snail Helix pomatia (pp. 604-610 in the 4th edition). The book > >is > > > >> out of print, but available used. > > > >> > > > >> Allen A. Smith, Ph.D. > > > >> Professor of Anatomy > > > >> Barry University > > > >> School of Graduate Medical Sciences > > > >> Podiatric Medicine and Surgery > > > >> Miami Shores, Florida ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 mfredrickson <@t> cohenderm.com Thu Sep 9 07:44:36 2004 From: mfredrickson <@t> cohenderm.com (Michael Fredrickson) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] CLIA regs Message-ID: <3D55809F621E7C438DA80098CDE3744D10C5E8@hub.cohenderm.com> For what it is worth, we were once sited by CLIA for lack of adequate tracking documentation for sending blocks and slides outside of our facility. We sent them via regular first class US mail and the block was never received at the other facility. It is our understanding that there should be a written policy regarding how these will be send, i.e. FedEx with tracking numbers etc., and how to follow up to ensure that materials were received at the other facility and a mechanism to ensure materials are returned. Michael Fredrickson, Technical Director Cohen Dermatopathology Newton, MA -----Original Message----- From: DDittus787@aol.com [mailto:DDittus787@aol.com] Sent: Wednesday, September 08, 2004 4:31 PM To: Dawn Cowie; histonet Subject: Re: [Histonet] CLIA regs Clia bows to the most stringent rule committee , so if the cap rules are more stringent than state or other , clia will go with cap. Dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peoshel <@t> wisc.edu Thu Sep 9 08:22:37 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Land snail dissection. In-Reply-To: <414020BF.22473.1CC595@localhost> References: <414020BF.22473.1CC595@localhost> Message-ID: Greg, Yep, we've got an Animal Care Committee here at Univ. Wisconsin, and all the rest. They -- and most such entities in the US -- just don't care about anything that doesn't have a backbone. Besides, MgCl2 and cooling *are* anesthesia for snails, as well as some crustiaceans at least (my own critters, if a I had a job doing critters, instead of running microscopes for biomedical types). MS-222 might work on aquatic snails, but not on land ones. CO2/CO might work on Pulmonates, come to think of it (land snails are Pulmonates). I have to agree with anesthetizing lobsters before dropping them in the boiling water, though -- I've never believed that they don't feel the boiling. Phil >Dear Phil (and others), > >All this talk of ripping open snail shells, injecting with MgCl2 and >heaven forbid, fixing alive! Does anyone else out there have to work >within the constraints of an Animal Care Committee? > >Some of you might be interested to hear that in a an academic >setting, all procedures carried out on research animals (be it >crustacean, fish, avian or mammalian) have to be approved under >guidelines for ethical and humane use and care of animals for >research. For instance, while I have no objection to boiling my >lobsters alive at home, at work, lobsters must me properly >anesthetized prior to humane euthanasia! It's a strange world we >work in by times! >Cheers! >Greg > >Date sent: Wed, 08 Sep 2004 15:38:30 -0500 >From: Philip Oshel >To: Histonet@Pathology.swmed.edu >Copies to: Subject: RE: [Histonet] Land >snail dissection. > >> Yep, CaCO3. >> Also yes. >> But, why bother with decalification? Just play crab and crack the >> shell open. The snail can be removed alive, if unhappy. I'd put it in >> MgCl2 first, then open the shell and remove the snail, cool it to >> further relax and anesthetize it, inject fixative into the mantle >> cavity (and possibly the hemocoel), then immersion-fix it. For >> sectioning, I'd dissect the snail into smaller pieces to insure >> proper fixation and infiltration paraffin -- they have a very tough, >> muscular foot, and the mantle can be as well. >> The radula that Gayle referred to earlier is mostly keratin, but many >> snails deposit CaCO3 or other minerals (including iron, if I remember >> right) in the tips of the radular teeth -- either way, it will cause >> grief when paraffin sectioning. It'd be better to carefully dissect >> away the radula and mount it whole -- whole mounts of radulae are >> used in molluc taxonomy anyway. If you do want to section the radula, >> you will need to plastic embed it. >> >> Phil >> >> >Out of curiosity - is the shell made of calcium? I'm asking because I >> >really don't know - not a trick question? Isn't a snail out of it's shell >> >just a slug? >> >(Now THAT is a joke.) >> > >> > >> >Jackie O' >> > >> > >> >Jacqueline M. O'Connor HT(ASCP) >> >Abbott Laboratories >> >Global Pharmaceutical Research and Development >> >Discovery Chemotherapeutics >> >847.938.4919 >> >Fax 847.938.3266 >> > >> > >> > >> > >> >Jose Luis Palazon Fernandez >> >Sent by: histonet-bounces@lists.utsouthwestern.edu >> >09/08/2004 11:56 AM >> > >> > >> > To: histonet@pathology.swmed.edu >> > cc: >> > Subject: Re: RE: [Histonet] Land snail dissection. >> > >> > >> >If the snail is small I recomend you to fix the whole snail and after >> >fixation, decalcify it with 10 % EDTA. then process and include the whole >> >snail. Hope this help >> > >> > >> > >> > >> > >> >El dia 08/09/2004 18:23 usted envio el siguiente mensaje: >> > >> >>Date: 8 de Septiembre de 2004 18:23:01 >> > >> >>From: "Smith, Allen" >> > >> >>Subject: RE: [Histonet] Land snail dissection. >> > >> >>To: gcallis@montana.edu, histonet@pathology.swmed.edu >> > >> >> >> > >> >> Many centuries ago, I forced a snail out of its shell by shredding a >> >pack of >> > >> >> cigarettes into a pint of water and dropping the snail into it. >> > >> >> Borradaile's THE INVERTEBRATA has instructions for dissecting the >> >European >> > >> >> garden snail Helix pomatia (pp. 604-610 in the 4th edition). The book >> >is >> > >> >> out of print, but available used. >> > >> >> >> > >> >> Allen A. Smith, Ph.D. >> > >> >> Professor of Anatomy >> > >> >> Barry University >> > >> >> School of Graduate Medical Sciences >> > >> >> Podiatric Medicine and Surgery >> > >> >> Miami Shores, Florida > > > >~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ >Greg Dobbin >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. -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From mcauliff <@t> umdnj.edu Thu Sep 9 11:35:09 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] specificity of Masson's trichrome In-Reply-To: <000d01c4961f$db08dc00$8a19fea9@walkercvzdlx4z> References: <000d01c4961f$db08dc00$8a19fea9@walkercvzdlx4z> Message-ID: <4140863D.1020301@umdnj.edu> Hi Carrie: While Masson's might (or might not) stain fibronectin and actin, no one (ie. reviewers of your work) will believe that it is specific without rigorous testing. If the results with Masson's do not agree with IHC, I cannot believe that anyone would throw out the IHC result in favor of the Masson's result. Geoff Walker/Langdon wrote: >Hi Histonetters, > >To clarify my question of the other day: is it true that Masson's trichrome could potentially stain fibronectin and/or smooth muscle actin, especially if it is being overexpressed? Is there a histochemical stain for fibronectin? We have already successfully done IHC,but are trying to reconcile data obtained with a few different approaches. We are mostly working in lung. > >Thanks to those who have already provided ideas! > >Carrie Langdon >Dept. of Pathology >McMaster University >Hamilton, Ontario >Canada >_______________________________________________ >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 akbitting <@t> geisinger.edu Thu Sep 9 08:42:30 2004 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] H pylori IHC controls Message-ID: Im still looking for a good H pylori control supplier for IHC. Any help? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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 convmcm <@t> cc.usu.edu Thu Sep 9 08:48:14 2004 From: convmcm <@t> cc.usu.edu (Connie McManus) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] IHC for Canine Distemper Virus In-Reply-To: <005801c4910c$100589d0$78065486@vdl220FAC> Message-ID: <000001c49673$a77a44b0$4a737b81@Cygnus> I get mine from VMRD in Pullman WA. Phone: 509/334-5815 website: www.vmrd.com They have a comprehensive online catalogue and have pretty good customer service. I also noticed USBiological has CDV also. Phone: 800/520-3011 website: www.usbio.net Also, CDV cross reacts with human measles virus. Have fun. (If it isn't fun, it isn't worth it) 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 Jan Shivers Sent: Thursday, September 02, 2004 9:44 AM To: histonet Subject: [Histonet] IHC for Canine Distemper Virus I would greatly appreciate any vendor source suggestions for Canine Distemper Virus antibody (for use in IHC on FFPE tissue). Thank you very much in advance. Jan Shivers IHC Supervisor Univ. of Minnesota Veterinary Diagnostic Lab 1333 Gortner Ave. St. Paul, MN 55108 shive003@umn.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From flemons <@t> bhset.org Thu Sep 9 09:11:22 2004 From: flemons <@t> bhset.org (Fran Lemons) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Vendors in Toronto Message-ID: Can someone out there provide me with a list of vendors for the S/C?? I know I saw one somewhere, but can't locate it. Thanks in advance..... Fran Walker From la.sebree <@t> hosp.wisc.edu Thu Sep 9 09:14:19 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] synaptophysin Message-ID: Annette, We retrieve our Synaptophysin in a decloaker for 2" in either our home made citrate buffer or Biocare's Bull's Eye and then stain on the NexES for 32". Ours isn't real intense either. Try decloaking for 3-4". This should help. 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: JAnorthernexp@cs.com [mailto:JAnorthernexp@cs.com] Sent: Wednesday, September 08, 2004 7:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] synaptophysin anyone using a ventana stainer and antibodies have a retrieval method using a decloker? Ours stains very light. Antibody is on for 32 mins we did have it set at 16mins. But increasing the time did not prove to make much difference. Any other ideas?? The tissue is formalin fixed and paraffin embedded. Thanks Annette _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mhorne <@t> upei.ca Thu Sep 9 09:18:34 2004 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Land snail dissection. In-Reply-To: References: <414020BF.22473.1CC595@localhost> Message-ID: <41403C09.25744.8BB06C@localhost> I used to use soda water to temporarily put Gammarus,a small crustacean, to sleep; I immersed them in it til they stopped twitching. I was told it was the CO2 that did it. Had to be careful not to overdo it and kill them, as I needed to recover them. As for lobster , I put them in the freezer for about 20 min or so til they hardly move , and then plunge them in boiling water head first. They supposedly die before waking up , so the theory goes. Makes me feel a little better and I don't lift the lid :-). Lobster goes well with garlic and butter too, Margaret Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada From cmconway <@t> usgs.gov Thu Sep 9 09:22:02 2004 From: cmconway <@t> usgs.gov (Carla M Conway) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] quantitative graded scale for IHC Message-ID: Hello, I would like to devise a quantitative graded scale for IHC results which would provide more info than just +/- staining. I have a reference where IHC staining intensity was rated (from pale pink to dark red) using alkaline phosphatase/Vector-Red, however we are using Envision+/AEC which seems to be an all or nothing (red or no red) chromogen with no gradation. We just received ImagePro image analysis software, so this may be another route to try. Thanks in advance for any comments or suggestions you may have. Sincerely, Carla Conway Western Fisheries Research Center Seattle, WA From JQB7 <@t> CDC.GOV Thu Sep 9 09:42:00 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Vendors in Toronto Message-ID: Go to www.nsh.org and click on NSH Convention. Then choose 2004 Exhibitors. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fran Lemons Sent: Thursday, September 09, 2004 10:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Vendors in Toronto Can someone out there provide me with a list of vendors for the S/C?? I know I saw one somewhere, but can't locate it. Thanks in advance..... Fran Walker _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peoshel <@t> wisc.edu Thu Sep 9 10:49:32 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Land snail dissection. In-Reply-To: <41403C09.25744.8BB06C@localhost> References: <414020BF.22473.1CC595@localhost> <41403C09.25744.8BB06C@localhost> Message-ID: Gammarus! One of my favorite critters -- did a lot of work on them at Memorial in St. John's NF. I didn't try soda water, but, yes it should work fine, and I agree, the CO2 should be what does the job. Lobster with garlic and butter ... mmm ... amazing how much lobster is left over to be "disposed of" when doing EM on the sinus gland. Phil >I used to use soda water to temporarily put Gammarus,a small >crustacean, to sleep; I immersed them in it til they stopped >twitching. I was told it was the CO2 that did it. Had to be careful >not to overdo it and kill them, as I needed to recover them. > > As for lobster , I put them in the freezer for about 20 min or >so til they hardly move , and then plunge them in boiling water >head first. They supposedly die before waking up , so the theory >goes. Makes me feel a little better and I don't lift the lid :-). > > Lobster goes well with garlic and butter too, > > > Margaret >Margaret Horne , >Histology Teaching Assistant, >Dept. of B.SC., >Atlantic Veterinary College, U.P.E.I., >550 University Ave., Charlottetown, >P.E.I., C1A 4P3 >Canada -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From dobbin <@t> upei.ca Thu Sep 9 10:54:09 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] quantitative graded scale for IHC In-Reply-To: Message-ID: <41405270.25853.DEE86B@localhost> Hi Carla, I certainly may be corrected by others on this but I personally don't think you will obtain statistically valid data by grading intensity of the reaction between cells or fields or sections. There is just so much room for subjectivity and for that matter, variability between runs or perhaps even between slides within runs! What you may want to try is calculating a ratio of positive to negative (perhaps pos. cells vs. neg. cells in a given area). For instance: place a grid with 50 or 100 intersects on the computer monitor, choose a microscopic field (in a random or systematic- random manner), and then at each intersect decide whether the reaction at that point is positive or negative. If at any particular intersect you have trouble to decide whether it is pos or neg, have a system that does not permit subjectivity, such as: for such points where the call could go either way, look at the space imediately to the left of the y-axis and above the x-axis and then make your decision. Determination of the number of fields you need to sample in order to obtain statistically valid data is very important and a matter for a statistician to explain and/or determine for you (which I most definately am not) unless you happen to have that training yourself. Another idea would be to have the image analysis software either measure the area of or count the number of pixels in a given area (field) that are "this red or redder". This involves choosing a relatively arbitrational threshold for the degree of redness that is to be measured or counted and applying the exact same setting to all other fields and sections in the study. My personal experience (using Bioquant NOVA) has been this sounds great in theory but was very difficult to pull off! The good old-fashioned counting was much more reliable and for that matter, easier to defend in presentation or publication. I am eager to hear other ideas or methods that might also work! Good luck. Greg To: Histonet@lists.utsouthwestern.edu From: "Carla M Conway" Date sent: Thu, 9 Sep 2004 07:22:02 -0700 Copies to: Subject: [Histonet] quantitative graded scale for IHC > Hello, > > I would like to devise a quantitative graded scale for IHC results which > would provide more info than just +/- staining. > I have a reference where IHC staining intensity was rated (from pale pink > to dark red) using alkaline phosphatase/Vector-Red, however we are using > Envision+/AEC which seems to be an all or nothing (red or no red) chromogen > with no gradation. We just received ImagePro image analysis software, so > this may be another route to try. Thanks in advance for any comments or > suggestions you may have. > > Sincerely, > > Carla Conway > Western Fisheries Research Center > Seattle, WA > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 froyer <@t> bitstream.net Thu Sep 9 11:05:12 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] How to relax a land =?windows-1252?Q?snail=85defi?= =?windows-1252?Q?nitively=2E?= In-Reply-To: References: Message-ID: <41407F38.3000502@bitstream.net> Subject: How to relax a land snail?definitively. Have you tried placing them in a warm bubble bath with subdued lighting, soft music playing, a little candle light, and a nice Merlot or Chablis to sip on? That always relaxes me... ~ Ford ;-) Ford M. Royer, MT(ASCP) Midwest Science and Biocenter (MSB) 6551 Jansen Ave. NE, Ste. 102 Albertville, MN 55301 (800) 745-4869 phone 763-497-6728 fax email Web URL Massimo wrote: >Torino 09 September 2004 >(ITALY) > >I would like to thank for your answers at my question about snail. >I also think to do something pleasant and to raise the scientific contents of someone answers by sending them an Italian recipe on snails with garlic. >Sorry in Italy we use the olive oil. Butter is very dangerous for the heart, the arteries ( just like the cigarettes smoke) and it also gets fat. >I am afraid but the recipe is in Italian. >But I?m sure if you enter a good Italian restaurant they can cook it. >But?remember? do not forget to bring with you the snails! > > >With my Best Regard > >Massimo Tosi > >Graduate Chemical Engineering > >PISA University >ITALY > > >LUMACHE CON L'AGLIO (SNAILS WITH GARLIC) > > >INGREDIENTI PER 4 PERSONE > >1Kg di lumache gi? spurgate (appena pulite vi consiglio di metterle per pi? di 2 ore al sole in un contenitore con abbondante sale sui bordi, le lumache infatti attirate dal sole e dal sale escono tutte fuori) >4 spicchi di aglio (GARLIC) >1 mazzetto di prezzemolo >olio extravergine di oliva (OLIVE OIL) >sale >pepe > >Lessate le lumache per 15 minuti, schiumando con un mestolo forato ("schiumarola" appunto). >A fine cottura, sciacquatele, rimettetele in tegame con acqua pulita leggermente salata e cuocete per altri 15 minuti. >A questo punto, sgocciolatele (conservate per? un po' d'acqua di cottura) e trasferitele in una casseruola, dove avete fatto soffriggere per poco tempo l'aglio, il prezzemolo e il pepe, cuocete per pochi minuti e aggiustate di sale, irrorate con l'acqua di cottura e servite in una zuppiera con del prezzemolo fresco. > >E adesso inizia lo spasso.... >buon appetito > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From AFeatherstone <@t> KaleidaHealth.Org Thu Sep 9 11:04:22 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Histotech position at Buffalo General Hospital Message-ID: We currently have 2 histotech positions open at Kaleida Health, Buffalo General Hospital in sunny Buffalo NY. Two shifts are available 4-12midnight and 8-4pm. These are full time, with full benefits and a great working environment. Please contact me if you are interested. Annette Featherstone HT/MLT Kaleida Health, Buffalo General Hospital 100 High St. Buffalo, NY 12403 716-859-2625 FAX: 716-859-1853 -----Original Message----- From: Greg Dobbin [mailto:dobbin@upei.ca] Sent: Thursday, September 09, 2004 08:54 To: Carla M Conway Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] quantitative graded scale for IHC Hi Carla, I certainly may be corrected by others on this but I personally don't think you will obtain statistically valid data by grading intensity of the reaction between cells or fields or sections. There is just so much room for subjectivity and for that matter, variability between runs or perhaps even between slides within runs! What you may want to try is calculating a ratio of positive to negative (perhaps pos. cells vs. neg. cells in a given area). For instance: place a grid with 50 or 100 intersects on the computer monitor, choose a microscopic field (in a random or systematic- random manner), and then at each intersect decide whether the reaction at that point is positive or negative. If at any particular intersect you have trouble to decide whether it is pos or neg, have a system that does not permit subjectivity, such as: for such points where the call could go either way, look at the space imediately to the left of the y-axis and above the x-axis and then make your decision. Determination of the number of fields you need to sample in order to obtain statistically valid data is very important and a matter for a statistician to explain and/or determine for you (which I most definately am not) unless you happen to have that training yourself. Another idea would be to have the image analysis software either measure the area of or count the number of pixels in a given area (field) that are "this red or redder". This involves choosing a relatively arbitrational threshold for the degree of redness that is to be measured or counted and applying the exact same setting to all other fields and sections in the study. My personal experience (using Bioquant NOVA) has been this sounds great in theory but was very difficult to pull off! The good old-fashioned counting was much more reliable and for that matter, easier to defend in presentation or publication. I am eager to hear other ideas or methods that might also work! Good luck. Greg To: Histonet@lists.utsouthwestern.edu From: "Carla M Conway" Date sent: Thu, 9 Sep 2004 07:22:02 -0700 Copies to: Subject: [Histonet] quantitative graded scale for IHC > Hello, > > I would like to devise a quantitative graded scale for IHC results which > would provide more info than just +/- staining. > I have a reference where IHC staining intensity was rated (from pale pink > to dark red) using alkaline phosphatase/Vector-Red, however we are using > Envision+/AEC which seems to be an all or nothing (red or no red) chromogen > with no gradation. We just received ImagePro image analysis software, so > this may be another route to try. Thanks in advance for any comments or > suggestions you may have. > > Sincerely, > > Carla Conway > Western Fisheries Research Center > Seattle, WA > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 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 gcallis <@t> montana.edu Thu Sep 9 11:23:32 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Vendors in Toronto In-Reply-To: References: Message-ID: <6.0.0.22.1.20040909102245.01b541f0@gemini.msu.montana.edu> Try NSH website, meeting information should have all this www.NSH.org, click on convention. At 08:11 AM 9/9/2004, you wrote: >Can someone out there provide me with a list of vendors for the S/C?? I >know I saw one somewhere, but can't locate it. >Thanks in advance..... >Fran Walker > > >_______________________________________________ >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 mhorne <@t> upei.ca Thu Sep 9 11:24:02 2004 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Land snail dissection. In-Reply-To: Message-ID: <41405971.8016.FE9066@localhost> Massimo, I am sorry that you do not often have the pleasure of eating lobster. If you are ever in Prince Edward Island , Canada , then I will feed you lobster. You might find it an interesting comparison with the Mediterranean lobster which has a different flavour and texture. Thank you for the recipe for escargot; I love them and always wanted to know how to cook them. I sent the recipe to my daughter who plans on going to Italy next summer on an archaeological dig and she sent me back a rough translation, which you can find in the attachment. Bon Appetit, Margaret Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada -------------- next part -------------- The following section of this message contains a file attachment prepared for transmission using the Internet MIME message format. If you are using Pegasus Mail, or any another MIME-compliant system, you should be able to save it or view it from within your mailer. If you cannot, please ask your system administrator for assistance. ---- File information ----------- File: snails.doc Date: 9 Sep 2004, 12:23 Size: 30720 bytes. Type: Unknown From Julie.Sanders <@t> med.va.gov Thu Sep 9 11:44:52 2004 From: Julie.Sanders <@t> med.va.gov (Julie.Sanders@med.va.gov) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] synaptophysin Message-ID: <457381D92B01BD44B21CF37CC02EBDFD28E935@vhacinexc2.v10.med.va.gov> Annette, When we had our Ventana NexEs we antigen retrieved in a pressure cooker....put two coplin jars with Cell Marque Trilogy in the cooker, place your slides in one of them and leave in pressure cooker 10 minutes after it reaches pressure. Relieve the steam from the cooker, remove the lid and place the slides in the other coplin jar of Trilogy for 10 minutes then rinse in APK was and place on the machine, 32 minute antibody time, with amplification. Julie Sanders Supervisor, Anatomic Pathlogy VAMC, Cincinnati, Ohio -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Thursday, September 09, 2004 12:37 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 10, Issue 12 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. specificity of Masson's trichrome (Walker/Langdon) 2. needed: used Fisher 166A or 166MP tissue processor (HCS) 3. PCNA - URINARY BLADDER (balajimr@drreddys.com) 4. Re:Splendore-Hoeppli phenomenon (renton louise mrs) 5. Shredded cigarettes OT (renton louise mrs) 6. p75 and Neuro D (Carl Hobbs) 7. How to relax a land snaildefinitively. (Massimo) 8. RE: synaptophysin (Featherstone, Annette) 9. RE: Land snail dissection. (Greg Dobbin) 10. RE: CLIA regs (Michael Fredrickson) 11. RE: Land snail dissection. (Philip Oshel) 12. Re: specificity of Masson's trichrome (Geoff McAuliffe) 13. H pylori IHC controls (Angela Bitting) 14. RE: IHC for Canine Distemper Virus (Connie McManus) 15. Vendors in Toronto (Fran Lemons) 16. RE: synaptophysin (Sebree Linda A.) 17. RE: Land snail dissection. (Margaret Horne) 18. quantitative graded scale for IHC (Carla M Conway) 19. RE: Vendors in Toronto (Bartlett, Jeanine) 20. RE: Land snail dissection. (Philip Oshel) 21. Re: quantitative graded scale for IHC (Greg Dobbin) 22. Re: How to relax a land snail?definitively. (Ford Royer) 23. Histotech position at Buffalo General Hospital (Featherstone, Annette) 24. Re: Vendors in Toronto (Gayle Callis) 25. RE: Land snail dissection. (Margaret Horne) ---------------------------------------------------------------------- Message: 1 Date: Wed, 8 Sep 2004 23:48:17 -0400 From: "Walker/Langdon" Subject: [Histonet] specificity of Masson's trichrome To: Message-ID: <000d01c4961f$db08dc00$8a19fea9@walkercvzdlx4z> Content-Type: text/plain; charset="iso-8859-1" Hi Histonetters, To clarify my question of the other day: is it true that Masson's trichrome could potentially stain fibronectin and/or smooth muscle actin, especially if it is being overexpressed? Is there a histochemical stain for fibronectin? We have already successfully done IHC,but are trying to reconcile data obtained with a few different approaches. We are mostly working in lung. Thanks to those who have already provided ideas! Carrie Langdon Dept. of Pathology McMaster University Hamilton, Ontario Canada ------------------------------ Message: 2 Date: Wed, 8 Sep 2004 21:18:46 -0700 From: "HCS" Subject: [Histonet] needed: used Fisher 166A or 166MP tissue processor To: "Histonet" Message-ID: <000701c49624$1a104740$6501a8c0@hp> Content-Type: text/plain; charset="iso-8859-1" Does anyone have an extra Fisher 166MP or 166A. Mine is beyond repair and would like to find a working unit. Please email directly. Hope the sales reps do not call, the budget does not allow for new units at this time. LeRoy Brown HT(ASCP) HTL HCS Everson, WA 98247 1-360-966-7300 ------------------------------ Message: 3 Date: Thu, 9 Sep 2004 12:04:53 +0530 From: balajimr@drreddys.com Subject: [Histonet] PCNA - URINARY BLADDER To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Dear Histonetters, It's nice to be back with Histosearch after a short break. Well, I am facing problem with PCN staining of urinary bladder of rat. We are using Zymed PCNA staining kit for the same. I am getting non specific staining where even the nucleus of the muscle layers are also staining. We are retrieving the antigen by heating the sections in Citrate buffer. But we are not having this problem in the intestines of same control animals. Does anybody have experience of PCNA staining with U. bladder so that they can advise me about this problem. Ofcourse we have increased the blocking time with serum upto 30 minutes unsuccessfully. Thanks in advance. Dr. M.R. Balaji Dept. Pre clinical safety evaluation, Discovery research, Dr. Reddys Laboratories ltd. Bollaram Road, Miyapur, Hyderabad, 500 049 Andhra Pradseh, INDIA Email - balajimr@drreddys.com Tel: 040- 23045439 - Ext.432. ------------------------------ Message: 4 Date: Thu, 09 Sep 2004 09:05:13 +0200 From: "renton louise mrs" Subject: [Histonet] Re:Splendore-Hoeppli phenomenon To: Terry.Marshall@rothgen.nhs.uk, histonet@lists.utsouthwestern.edu Message-ID: <1094713513.8dd9db60rentonlf@bru.wits.ac.za> Content-Type: text/plain; charset="UTF-8" Now, if we could only use that in the lyrics of a song along the lines of: "lung.... has has Hoeppli-splendore thing" cheers -----Original Message----- From: "Marshall Terry Dr, Consultant Histopathologist" To: "Angela Bitting" , Date: Wed, 8 Sep 2004 14:19:44 +0100 Subject: RE: [Histonet] Modified acid fast stain for actinomyces No modification is needed. Neither does it stain the Actinomyces. Rather, the club shaped ends of the filaments, being an antigen-antibody complex - known as Splendore-Hoeppli phenomenon - are stained red. They are not necessarily present. 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: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: 08 September 2004 13:53 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Modified acid fast stain for actinomyces Anyone have a recipe for this stain? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? ------------------------------ Message: 5 Date: Thu, 09 Sep 2004 09:07:28 +0200 From: "renton louise mrs" Subject: [Histonet] Shredded cigarettes OT To: histonet@lists.utsouthwestern.edu Message-ID: <1094713648.8dd9db60rentonlf@bru.wits.ac.za> Content-Type: text/plain; charset="UTF-8" Tobacco powder is still widely used by gardeners as a "natural/organic" insecticide -----Original Message----- From: Geoff McAuliffe To: Gayle Callis Date: Wed, 08 Sep 2004 15:36:28 -0700 Subject: Re: [Histonet] Shredded cigarettes for Land snail dissection. I think so, I think it is the nicotine that is the narcotic here. Geoff Gayle Callis wrote: > At the risk of too much humor, would chewing tobacco (not used, > please!) do the job instead of shredding cigarettes. A scientific use > for SKOL!! > > At 10:23 AM 9/8/2004, you wrote: > >> Many centuries ago, I forced a snail out of its shell by shredding a >> pack of >> cigarettes into a pint of water and dropping the snail into it. >> Borradaile's THE INVERTEBRATA has instructions for dissecting the >> European >> garden snail Helix pomatia (pp. 604-610 in the 4th edition). The >> book is >> out of print, but available used. >> >> Allen A. Smith, Ph.D. >> Professor of Anatomy >> Barry University >> School of Graduate Medical Sciences >> Podiatric Medicine and Surgery >> Miami Shores, Florida > > > 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 > -- -- ********************************************** 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 Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? ------------------------------ Message: 6 Date: Thu, 9 Sep 2004 08:48:06 +0100 From: "Carl Hobbs" Subject: [Histonet] p75 and Neuro D To: Message-ID: <001701c49641$5805c3f0$e8345c9f@Carlos> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Be most grateful if anyone out there can recommend antibodies against those two proteins, that work in mouse pwx sections. I have got Chemicon's p75 ( MAB 390). The p75 is the low affinity neurotrophin receptor. I have searched but I obviously am reluctant to buy unless confirmed that they work in pwax. Thank you ------------------------------ Message: 7 Date: Thu, 9 Sep 2004 12:57:20 +0200 From: "Massimo" Subject: [Histonet] How to relax a land snaildefinitively. To: "histonet histonet" Cc: Dawn Truscott , Carol McCollough Message-ID: Content-Type: text/plain; charset=iso-8859-1 Torino 09 September 2004 (ITALY) I would like to thank for your answers at my question about snail. I also think to do something pleasant and to raise the scientific contents of someone answers by sending them an Italian recipe on snails with garlic. Sorry in Italy we use the olive oil. Butter is very dangerous for the heart, the arteries ( just like the cigarettes smoke) and it also gets fat. I am afraid but the recipe is in Italian. But I'm sure if you enter a good Italian restaurant they can cook it. But...remember... do not forget to bring with you the snails! With my Best Regard Massimo Tosi Graduate Chemical Engineering PISA University ITALY LUMACHE CON L'AGLIO (SNAILS WITH GARLIC) INGREDIENTI PER 4 PERSONE 1Kg di lumache gi? spurgate (appena pulite vi consiglio di metterle per pi? di 2 ore al sole in un contenitore con abbondante sale sui bordi, le lumache infatti attirate dal sole e dal sale escono tutte fuori) 4 spicchi di aglio (GARLIC) 1 mazzetto di prezzemolo olio extravergine di oliva (OLIVE OIL) sale pepe Lessate le lumache per 15 minuti, schiumando con un mestolo forato ("schiumarola" appunto). A fine cottura, sciacquatele, rimettetele in tegame con acqua pulita leggermente salata e cuocete per altri 15 minuti. A questo punto, sgocciolatele (conservate per? un po' d'acqua di cottura) e trasferitele in una casseruola, dove avete fatto soffriggere per poco tempo l'aglio, il prezzemolo e il pepe, cuocete per pochi minuti e aggiustate di sale, irrorate con l'acqua di cottura e servite in una zuppiera con del prezzemolo fresco. E adesso inizia lo spasso.... buon appetito ------------------------------ Message: 8 Date: Thu, 9 Sep 2004 07:14:17 -0400 From: "Featherstone, Annette" Subject: RE: [Histonet] synaptophysin To: 'Joe Nocito' , JAnorthernexp@cs.com, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" We antigen retrieve our synaptophysin in Citrate Buffer ph6.0 in a steamer. We set the steamer with the citrate in it for 70 minutes, at 20 minutes left we add the slides. After it steams for the final 20 minutes we let it cool for 20 minutes. We add amplifier and blocker too. Is your primary dilution correct? Annette Featherstone HT/MLT Kaleida Health. -----Original Message----- From: Joe Nocito [mailto:jnocito@satx.rr.com] Sent: Wednesday, September 08, 2004 22:02 To: JAnorthernexp@cs.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] synaptophysin Annette, did you try increasing the time in cell conditioner? Have you tried to amplify it? Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: To: Sent: Wednesday, September 08, 2004 7:06 PM Subject: [Histonet] synaptophysin > anyone using a ventana stainer and antibodies have a retrieval method using a > decloker? Ours stains very light. Antibody is on for 32 mins we did have it > set at 16mins. But increasing the time did not prove to make much > difference. Any other ideas?? The tissue is formalin fixed and paraffin embedded. > Thanks > > Annette > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. ------------------------------ Message: 9 Date: Thu, 9 Sep 2004 09:22:07 ADT From: "Greg Dobbin" Subject: RE: [Histonet] Land snail dissection. To: Histonet@Pathology.swmed.edu Message-ID: <414020BF.22473.1CC595@localhost> Content-Type: text/plain; charset=US-ASCII Dear Phil (and others), All this talk of ripping open snail shells, injecting with MgCl2 and heaven forbid, fixing alive! Does anyone else out there have to work within the constraints of an Animal Care Committee? Some of you might be interested to hear that in a an academic setting, all procedures carried out on research animals (be it crustacean, fish, avian or mammalian) have to be approved under guidelines for ethical and humane use and care of animals for research. For instance, while I have no objection to boiling my lobsters alive at home, at work, lobsters must me properly anesthetized prior to humane euthanasia! It's a strange world we work in by times! Cheers! Greg Date sent: Wed, 08 Sep 2004 15:38:30 -0500 From: Philip Oshel To: Histonet@Pathology.swmed.edu Copies to: Subject: RE: [Histonet] Land snail dissection. > Yep, CaCO3. > Also yes. > But, why bother with decalification? Just play crab and crack the > shell open. The snail can be removed alive, if unhappy. I'd put it in > MgCl2 first, then open the shell and remove the snail, cool it to > further relax and anesthetize it, inject fixative into the mantle > cavity (and possibly the hemocoel), then immersion-fix it. For > sectioning, I'd dissect the snail into smaller pieces to insure > proper fixation and infiltration paraffin -- they have a very tough, > muscular foot, and the mantle can be as well. > The radula that Gayle referred to earlier is mostly keratin, but many > snails deposit CaCO3 or other minerals (including iron, if I remember > right) in the tips of the radular teeth -- either way, it will cause > grief when paraffin sectioning. It'd be better to carefully dissect > away the radula and mount it whole -- whole mounts of radulae are > used in molluc taxonomy anyway. If you do want to section the radula, > you will need to plastic embed it. > > Phil > > >Out of curiosity - is the shell made of calcium? I'm asking because I > >really don't know - not a trick question? Isn't a snail out of it's shell > >just a slug? > >(Now THAT is a joke.) > > > > > >Jackie O' > > > > > >Jacqueline M. O'Connor HT(ASCP) > >Abbott Laboratories > >Global Pharmaceutical Research and Development > >Discovery Chemotherapeutics > >847.938.4919 > >Fax 847.938.3266 > > > > > > > > > >Jose Luis Palazon Fernandez > >Sent by: histonet-bounces@lists.utsouthwestern.edu > >09/08/2004 11:56 AM > > > > > > To: histonet@pathology.swmed.edu > > cc: > > Subject: Re: RE: [Histonet] Land snail dissection. > > > > > >If the snail is small I recomend you to fix the whole snail and after > >fixation, decalcify it with 10 % EDTA. then process and include the whole > >snail. Hope this help > > > > > > > > > > > >El dia 08/09/2004 18:23 usted envio el siguiente mensaje: > > > >>Date: 8 de Septiembre de 2004 18:23:01 > > > >>From: "Smith, Allen" > > > >>Subject: RE: [Histonet] Land snail dissection. > > > >>To: gcallis@montana.edu, histonet@pathology.swmed.edu > > > >> > > > >> Many centuries ago, I forced a snail out of its shell by shredding a > >pack of > > > >> cigarettes into a pint of water and dropping the snail into it. > > > >> Borradaile's THE INVERTEBRATA has instructions for dissecting the > >European > > > >> garden snail Helix pomatia (pp. 604-610 in the 4th edition). The book > >is > > > >> out of print, but available used. > > > >> > > > >> Allen A. Smith, Ph.D. > > > >> Professor of Anatomy > > > >> Barry University > > > >> School of Graduate Medical Sciences > > > >> Podiatric Medicine and Surgery > > > >> Miami Shores, Florida ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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. ------------------------------ Message: 10 Date: Thu, 9 Sep 2004 08:44:36 -0400 From: "Michael Fredrickson" Subject: RE: [Histonet] CLIA regs To: "Dawn Cowie" , "histonet" Message-ID: <3D55809F621E7C438DA80098CDE3744D10C5E8@hub.cohenderm.com> Content-Type: text/plain; charset="iso-8859-1" For what it is worth, we were once sited by CLIA for lack of adequate tracking documentation for sending blocks and slides outside of our facility. We sent them via regular first class US mail and the block was never received at the other facility. It is our understanding that there should be a written policy regarding how these will be send, i.e. FedEx with tracking numbers etc., and how to follow up to ensure that materials were received at the other facility and a mechanism to ensure materials are returned. Michael Fredrickson, Technical Director Cohen Dermatopathology Newton, MA -----Original Message----- From: DDittus787@aol.com [mailto:DDittus787@aol.com] Sent: Wednesday, September 08, 2004 4:31 PM To: Dawn Cowie; histonet Subject: Re: [Histonet] CLIA regs Clia bows to the most stringent rule committee , so if the cap rules are more stringent than state or other , clia will go with cap. Dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Thu, 09 Sep 2004 08:22:37 -0500 From: Philip Oshel Subject: RE: [Histonet] Land snail dissection. To: Histonet@Pathology.swmed.edu Message-ID: Content-Type: text/plain; format=flowed; charset=us-ascii Greg, Yep, we've got an Animal Care Committee here at Univ. Wisconsin, and all the rest. They -- and most such entities in the US -- just don't care about anything that doesn't have a backbone. Besides, MgCl2 and cooling *are* anesthesia for snails, as well as some crustiaceans at least (my own critters, if a I had a job doing critters, instead of running microscopes for biomedical types). MS-222 might work on aquatic snails, but not on land ones. CO2/CO might work on Pulmonates, come to think of it (land snails are Pulmonates). I have to agree with anesthetizing lobsters before dropping them in the boiling water, though -- I've never believed that they don't feel the boiling. Phil >Dear Phil (and others), > >All this talk of ripping open snail shells, injecting with MgCl2 and >heaven forbid, fixing alive! Does anyone else out there have to work >within the constraints of an Animal Care Committee? > >Some of you might be interested to hear that in a an academic >setting, all procedures carried out on research animals (be it >crustacean, fish, avian or mammalian) have to be approved under >guidelines for ethical and humane use and care of animals for >research. For instance, while I have no objection to boiling my >lobsters alive at home, at work, lobsters must me properly >anesthetized prior to humane euthanasia! It's a strange world we >work in by times! >Cheers! >Greg > >Date sent: Wed, 08 Sep 2004 15:38:30 -0500 >From: Philip Oshel >To: Histonet@Pathology.swmed.edu >Copies to: Subject: RE: [Histonet] Land >snail dissection. > >> Yep, CaCO3. >> Also yes. >> But, why bother with decalification? Just play crab and crack the >> shell open. The snail can be removed alive, if unhappy. I'd put it in >> MgCl2 first, then open the shell and remove the snail, cool it to >> further relax and anesthetize it, inject fixative into the mantle >> cavity (and possibly the hemocoel), then immersion-fix it. For >> sectioning, I'd dissect the snail into smaller pieces to insure >> proper fixation and infiltration paraffin -- they have a very tough, >> muscular foot, and the mantle can be as well. >> The radula that Gayle referred to earlier is mostly keratin, but many >> snails deposit CaCO3 or other minerals (including iron, if I remember >> right) in the tips of the radular teeth -- either way, it will cause >> grief when paraffin sectioning. It'd be better to carefully dissect >> away the radula and mount it whole -- whole mounts of radulae are >> used in molluc taxonomy anyway. If you do want to section the radula, >> you will need to plastic embed it. >> >> Phil >> >> >Out of curiosity - is the shell made of calcium? I'm asking because I >> >really don't know - not a trick question? Isn't a snail out of it's shell >> >just a slug? >> >(Now THAT is a joke.) >> > >> > >> >Jackie O' >> > >> > >> >Jacqueline M. O'Connor HT(ASCP) >> >Abbott Laboratories >> >Global Pharmaceutical Research and Development >> >Discovery Chemotherapeutics >> >847.938.4919 >> >Fax 847.938.3266 >> > >> > >> > >> > >> >Jose Luis Palazon Fernandez >> >Sent by: histonet-bounces@lists.utsouthwestern.edu >> >09/08/2004 11:56 AM >> > >> > >> > To: histonet@pathology.swmed.edu >> > cc: >> > Subject: Re: RE: [Histonet] Land snail dissection. >> > >> > >> >If the snail is small I recomend you to fix the whole snail and after >> >fixation, decalcify it with 10 % EDTA. then process and include the whole >> >snail. Hope this help >> > >> > >> > >> > >> > >> >El dia 08/09/2004 18:23 usted envio el siguiente mensaje: >> > >> >>Date: 8 de Septiembre de 2004 18:23:01 >> > >> >>From: "Smith, Allen" >> > >> >>Subject: RE: [Histonet] Land snail dissection. >> > >> >>To: gcallis@montana.edu, histonet@pathology.swmed.edu >> > >> >> >> > >> >> Many centuries ago, I forced a snail out of its shell by shredding a >> >pack of >> > >> >> cigarettes into a pint of water and dropping the snail into it. >> > >> >> Borradaile's THE INVERTEBRATA has instructions for dissecting the >> >European >> > >> >> garden snail Helix pomatia (pp. 604-610 in the 4th edition). The book >> >is >> > >> >> out of print, but available used. >> > >> >> >> > >> >> Allen A. Smith, Ph.D. >> > >> >> Professor of Anatomy >> > >> >> Barry University >> > >> >> School of Graduate Medical Sciences >> > >> >> Podiatric Medicine and Surgery >> > >> >> Miami Shores, Florida > > > >~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ >Greg Dobbin >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. -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) ------------------------------ Message: 12 Date: Thu, 09 Sep 2004 09:35:09 -0700 From: Geoff McAuliffe Subject: Re: [Histonet] specificity of Masson's trichrome To: Walker/Langdon Cc: histonet@lists.utsouthwestern.edu Message-ID: <4140863D.1020301@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Hi Carrie: While Masson's might (or might not) stain fibronectin and actin, no one (ie. reviewers of your work) will believe that it is specific without rigorous testing. If the results with Masson's do not agree with IHC, I cannot believe that anyone would throw out the IHC result in favor of the Masson's result. Geoff Walker/Langdon wrote: >Hi Histonetters, > >To clarify my question of the other day: is it true that Masson's trichrome could potentially stain fibronectin and/or smooth muscle actin, especially if it is being overexpressed? Is there a histochemical stain for fibronectin? We have already successfully done IHC,but are trying to reconcile data obtained with a few different approaches. We are mostly working in lung. > >Thanks to those who have already provided ideas! > >Carrie Langdon >Dept. of Pathology >McMaster University >Hamilton, Ontario >Canada >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** ------------------------------ Message: 13 Date: Thu, 09 Sep 2004 09:42:30 -0400 From: "Angela Bitting" Subject: [Histonet] H pylori IHC controls To: Message-ID: Content-Type: text/plain; charset=US-ASCII Im still looking for a good H pylori control supplier for IHC. Any help? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 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. ------------------------------ Message: 14 Date: Thu, 9 Sep 2004 07:48:14 -0600 From: "Connie McManus" Subject: RE: [Histonet] IHC for Canine Distemper Virus To: "'Jan Shivers'" , "'histonet'" Message-ID: <000001c49673$a77a44b0$4a737b81@Cygnus> Content-Type: text/plain; charset="us-ascii" I get mine from VMRD in Pullman WA. Phone: 509/334-5815 website: www.vmrd.com They have a comprehensive online catalogue and have pretty good customer service. I also noticed USBiological has CDV also. Phone: 800/520-3011 website: www.usbio.net Also, CDV cross reacts with human measles virus. Have fun. (If it isn't fun, it isn't worth it) 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 Jan Shivers Sent: Thursday, September 02, 2004 9:44 AM To: histonet Subject: [Histonet] IHC for Canine Distemper Virus I would greatly appreciate any vendor source suggestions for Canine Distemper Virus antibody (for use in IHC on FFPE tissue). Thank you very much in advance. Jan Shivers IHC Supervisor Univ. of Minnesota Veterinary Diagnostic Lab 1333 Gortner Ave. St. Paul, MN 55108 shive003@umn.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Thu, 09 Sep 2004 10:11:22 -0400 From: "Fran Lemons" Subject: [Histonet] Vendors in Toronto To: Message-ID: Content-Type: text/plain; charset=US-ASCII Can someone out there provide me with a list of vendors for the S/C?? I know I saw one somewhere, but can't locate it. Thanks in advance..... Fran Walker ------------------------------ Message: 16 Date: Thu, 9 Sep 2004 09:14:19 -0500 From: "Sebree Linda A." Subject: RE: [Histonet] synaptophysin To: , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Annette, We retrieve our Synaptophysin in a decloaker for 2" in either our home made citrate buffer or Biocare's Bull's Eye and then stain on the NexES for 32". Ours isn't real intense either. Try decloaking for 3-4". This should help. 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: JAnorthernexp@cs.com [mailto:JAnorthernexp@cs.com] Sent: Wednesday, September 08, 2004 7:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] synaptophysin anyone using a ventana stainer and antibodies have a retrieval method using a decloker? Ours stains very light. Antibody is on for 32 mins we did have it set at 16mins. But increasing the time did not prove to make much difference. Any other ideas?? The tissue is formalin fixed and paraffin embedded. Thanks Annette _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Thu, 9 Sep 2004 11:18:34 ADT From: "Margaret Horne" Subject: RE: [Histonet] Land snail dissection. To: Histonet@Pathology.swmed.edu, Philip Oshel Message-ID: <41403C09.25744.8BB06C@localhost> Content-Type: text/plain; charset=US-ASCII I used to use soda water to temporarily put Gammarus,a small crustacean, to sleep; I immersed them in it til they stopped twitching. I was told it was the CO2 that did it. Had to be careful not to overdo it and kill them, as I needed to recover them. As for lobster , I put them in the freezer for about 20 min or so til they hardly move , and then plunge them in boiling water head first. They supposedly die before waking up , so the theory goes. Makes me feel a little better and I don't lift the lid :-). Lobster goes well with garlic and butter too, Margaret Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada ------------------------------ Message: 18 Date: Thu, 9 Sep 2004 07:22:02 -0700 From: "Carla M Conway" Subject: [Histonet] quantitative graded scale for IHC To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Hello, I would like to devise a quantitative graded scale for IHC results which would provide more info than just +/- staining. I have a reference where IHC staining intensity was rated (from pale pink to dark red) using alkaline phosphatase/Vector-Red, however we are using Envision+/AEC which seems to be an all or nothing (red or no red) chromogen with no gradation. We just received ImagePro image analysis software, so this may be another route to try. Thanks in advance for any comments or suggestions you may have. Sincerely, Carla Conway Western Fisheries Research Center Seattle, WA ------------------------------ Message: 19 Date: Thu, 9 Sep 2004 10:42:00 -0400 From: "Bartlett, Jeanine" Subject: RE: [Histonet] Vendors in Toronto To: "Fran Lemons" , Message-ID: Content-Type: text/plain; charset="US-ASCII" Go to www.nsh.org and click on NSH Convention. Then choose 2004 Exhibitors. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fran Lemons Sent: Thursday, September 09, 2004 10:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Vendors in Toronto Can someone out there provide me with a list of vendors for the S/C?? I know I saw one somewhere, but can't locate it. Thanks in advance..... Fran Walker _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 20 Date: Thu, 09 Sep 2004 10:49:32 -0500 From: Philip Oshel Subject: RE: [Histonet] Land snail dissection. To: Margaret Horne Cc: Histonet@Pathology.swmed.edu Message-ID: Content-Type: text/plain; format=flowed; charset=us-ascii Gammarus! One of my favorite critters -- did a lot of work on them at Memorial in St. John's NF. I didn't try soda water, but, yes it should work fine, and I agree, the CO2 should be what does the job. Lobster with garlic and butter ... mmm ... amazing how much lobster is left over to be "disposed of" when doing EM on the sinus gland. Phil >I used to use soda water to temporarily put Gammarus,a small >crustacean, to sleep; I immersed them in it til they stopped >twitching. I was told it was the CO2 that did it. Had to be careful >not to overdo it and kill them, as I needed to recover them. > > As for lobster , I put them in the freezer for about 20 min or >so til they hardly move , and then plunge them in boiling water >head first. They supposedly die before waking up , so the theory >goes. Makes me feel a little better and I don't lift the lid :-). > > Lobster goes well with garlic and butter too, > > > Margaret >Margaret Horne , >Histology Teaching Assistant, >Dept. of B.SC., >Atlantic Veterinary College, U.P.E.I., >550 University Ave., Charlottetown, >P.E.I., C1A 4P3 >Canada -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) ------------------------------ Message: 21 Date: Thu, 9 Sep 2004 12:54:09 ADT From: "Greg Dobbin" Subject: Re: [Histonet] quantitative graded scale for IHC To: "Carla M Conway" Cc: Histonet@lists.utsouthwestern.edu Message-ID: <41405270.25853.DEE86B@localhost> Content-Type: text/plain; charset=US-ASCII Hi Carla, I certainly may be corrected by others on this but I personally don't think you will obtain statistically valid data by grading intensity of the reaction between cells or fields or sections. There is just so much room for subjectivity and for that matter, variability between runs or perhaps even between slides within runs! What you may want to try is calculating a ratio of positive to negative (perhaps pos. cells vs. neg. cells in a given area). For instance: place a grid with 50 or 100 intersects on the computer monitor, choose a microscopic field (in a random or systematic- random manner), and then at each intersect decide whether the reaction at that point is positive or negative. If at any particular intersect you have trouble to decide whether it is pos or neg, have a system that does not permit subjectivity, such as: for such points where the call could go either way, look at the space imediately to the left of the y-axis and above the x-axis and then make your decision. Determination of the number of fields you need to sample in order to obtain statistically valid data is very important and a matter for a statistician to explain and/or determine for you (which I most definately am not) unless you happen to have that training yourself. Another idea would be to have the image analysis software either measure the area of or count the number of pixels in a given area (field) that are "this red or redder". This involves choosing a relatively arbitrational threshold for the degree of redness that is to be measured or counted and applying the exact same setting to all other fields and sections in the study. My personal experience (using Bioquant NOVA) has been this sounds great in theory but was very difficult to pull off! The good old-fashioned counting was much more reliable and for that matter, easier to defend in presentation or publication. I am eager to hear other ideas or methods that might also work! Good luck. Greg To: Histonet@lists.utsouthwestern.edu From: "Carla M Conway" Date sent: Thu, 9 Sep 2004 07:22:02 -0700 Copies to: Subject: [Histonet] quantitative graded scale for IHC > Hello, > > I would like to devise a quantitative graded scale for IHC results which > would provide more info than just +/- staining. > I have a reference where IHC staining intensity was rated (from pale pink > to dark red) using alkaline phosphatase/Vector-Red, however we are using > Envision+/AEC which seems to be an all or nothing (red or no red) chromogen > with no gradation. We just received ImagePro image analysis software, so > this may be another route to try. Thanks in advance for any comments or > suggestions you may have. > > Sincerely, > > Carla Conway > Western Fisheries Research Center > Seattle, WA > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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. ------------------------------ Message: 22 Date: Thu, 09 Sep 2004 11:05:12 -0500 From: Ford Royer Subject: Re: [Histonet] How to relax a land snail?definitively. To: histonet histonet Message-ID: <41407F38.3000502@bitstream.net> Content-Type: text/plain; charset=windows-1252; format=flowed Subject: How to relax a land snail...definitively. Have you tried placing them in a warm bubble bath with subdued lighting, soft music playing, a little candle light, and a nice Merlot or Chablis to sip on? That always relaxes me... ~ Ford ;-) Ford M. Royer, MT(ASCP) Midwest Science and Biocenter (MSB) 6551 Jansen Ave. NE, Ste. 102 Albertville, MN 55301 (800) 745-4869 phone 763-497-6728 fax email Web URL Massimo wrote: >Torino 09 September 2004 >(ITALY) > >I would like to thank for your answers at my question about snail. >I also think to do something pleasant and to raise the scientific contents of someone answers by sending them an Italian recipe on snails with garlic. >Sorry in Italy we use the olive oil. Butter is very dangerous for the heart, the arteries ( just like the cigarettes smoke) and it also gets fat. >I am afraid but the recipe is in Italian. >But I'm sure if you enter a good Italian restaurant they can cook it. >But...remember... do not forget to bring with you the snails! > > >With my Best Regard > >Massimo Tosi > >Graduate Chemical Engineering > >PISA University >ITALY > > >LUMACHE CON L'AGLIO (SNAILS WITH GARLIC) > > >INGREDIENTI PER 4 PERSONE > >1Kg di lumache gi? spurgate (appena pulite vi consiglio di metterle per pi? di 2 ore al sole in un contenitore con abbondante sale sui bordi, le lumache infatti attirate dal sole e dal sale escono tutte fuori) >4 spicchi di aglio (GARLIC) >1 mazzetto di prezzemolo >olio extravergine di oliva (OLIVE OIL) >sale >pepe > >Lessate le lumache per 15 minuti, schiumando con un mestolo forato ("schiumarola" appunto). >A fine cottura, sciacquatele, rimettetele in tegame con acqua pulita leggermente salata e cuocete per altri 15 minuti. >A questo punto, sgocciolatele (conservate per? un po' d'acqua di cottura) e trasferitele in una casseruola, dove avete fatto soffriggere per poco tempo l'aglio, il prezzemolo e il pepe, cuocete per pochi minuti e aggiustate di sale, irrorate con l'acqua di cottura e servite in una zuppiera con del prezzemolo fresco. > >E adesso inizia lo spasso.... >buon appetito > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > ------------------------------ Message: 23 Date: Thu, 9 Sep 2004 12:04:22 -0400 From: "Featherstone, Annette" Subject: [Histonet] Histotech position at Buffalo General Hospital To: 'Greg Dobbin' , Carla M Conway Cc: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" We currently have 2 histotech positions open at Kaleida Health, Buffalo General Hospital in sunny Buffalo NY. Two shifts are available 4-12midnight and 8-4pm. These are full time, with full benefits and a great working environment. Please contact me if you are interested. Annette Featherstone HT/MLT Kaleida Health, Buffalo General Hospital 100 High St. Buffalo, NY 12403 716-859-2625 FAX: 716-859-1853 -----Original Message----- From: Greg Dobbin [mailto:dobbin@upei.ca] Sent: Thursday, September 09, 2004 08:54 To: Carla M Conway Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] quantitative graded scale for IHC Hi Carla, I certainly may be corrected by others on this but I personally don't think you will obtain statistically valid data by grading intensity of the reaction between cells or fields or sections. There is just so much room for subjectivity and for that matter, variability between runs or perhaps even between slides within runs! What you may want to try is calculating a ratio of positive to negative (perhaps pos. cells vs. neg. cells in a given area). For instance: place a grid with 50 or 100 intersects on the computer monitor, choose a microscopic field (in a random or systematic- random manner), and then at each intersect decide whether the reaction at that point is positive or negative. If at any particular intersect you have trouble to decide whether it is pos or neg, have a system that does not permit subjectivity, such as: for such points where the call could go either way, look at the space imediately to the left of the y-axis and above the x-axis and then make your decision. Determination of the number of fields you need to sample in order to obtain statistically valid data is very important and a matter for a statistician to explain and/or determine for you (which I most definately am not) unless you happen to have that training yourself. Another idea would be to have the image analysis software either measure the area of or count the number of pixels in a given area (field) that are "this red or redder". This involves choosing a relatively arbitrational threshold for the degree of redness that is to be measured or counted and applying the exact same setting to all other fields and sections in the study. My personal experience (using Bioquant NOVA) has been this sounds great in theory but was very difficult to pull off! The good old-fashioned counting was much more reliable and for that matter, easier to defend in presentation or publication. I am eager to hear other ideas or methods that might also work! Good luck. Greg To: Histonet@lists.utsouthwestern.edu From: "Carla M Conway" Date sent: Thu, 9 Sep 2004 07:22:02 -0700 Copies to: Subject: [Histonet] quantitative graded scale for IHC > Hello, > > I would like to devise a quantitative graded scale for IHC results which > would provide more info than just +/- staining. > I have a reference where IHC staining intensity was rated (from pale pink > to dark red) using alkaline phosphatase/Vector-Red, however we are using > Envision+/AEC which seems to be an all or nothing (red or no red) chromogen > with no gradation. We just received ImagePro image analysis software, so > this may be another route to try. Thanks in advance for any comments or > suggestions you may have. > > Sincerely, > > Carla Conway > Western Fisheries Research Center > Seattle, WA > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. ------------------------------ Message: 24 Date: Thu, 09 Sep 2004 10:23:32 -0600 From: Gayle Callis Subject: Re: [Histonet] Vendors in Toronto To: "Fran Lemons" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20040909102245.01b541f0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Try NSH website, meeting information should have all this www.NSH.org, click on convention. At 08:11 AM 9/9/2004, you wrote: >Can someone out there provide me with a list of vendors for the S/C?? I >know I saw one somewhere, but can't locate it. >Thanks in advance..... >Fran Walker > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 25 Date: Thu, 9 Sep 2004 13:24:02 ADT From: "Margaret Horne" Subject: RE: [Histonet] Land snail dissection. To: "Massimo" Cc: Histonet@Pathology.swmed.edu Message-ID: <41405971.8016.FE9066@localhost> Content-Type: text/plain; charset="us-ascii" Massimo, I am sorry that you do not often have the pleasure of eating lobster. If you are ever in Prince Edward Island , Canada , then I will feed you lobster. You might find it an interesting comparison with the Mediterranean lobster which has a different flavour and texture. Thank you for the recipe for escargot; I love them and always wanted to know how to cook them. I sent the recipe to my daughter who plans on going to Italy next summer on an archaeological dig and she sent me back a rough translation, which you can find in the attachment. Bon Appetit, Margaret Margaret Horne , Histology Teaching Assistant, Dept. of B.SC., Atlantic Veterinary College, U.P.E.I., 550 University Ave., Charlottetown, P.E.I., C1A 4P3 Canada -------------- next part -------------- The following section of this message contains a file attachment prepared for transmission using the Internet MIME message format. If you are using Pegasus Mail, or any another MIME-compliant system, you should be able to save it or view it from within your mailer. If you cannot, please ask your system administrator for assistance. ---- File information ----------- File: snails.doc Date: 9 Sep 2004, 12:23 Size: 30720 bytes. Type: Unknown ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 10, Issue 12 **************************************** From thoward <@t> unm.edu Thu Sep 9 11:48:20 2004 From: thoward <@t> unm.edu (Tamara Howard) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] RE: Land snail dissection Message-ID: For what it is worth, MS222 will work on pulmonates - you just have to get them to stretch out and dip their foot into the solution. Subdued lighting helps...never tried the chablis idea. Cultured cells are soooooooooooooo much easier to deal with than inverts! |--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward@unm.edu |--------------------------------------------------| From chiggerson <@t> memhosp.com Thu Sep 9 11:52:54 2004 From: chiggerson <@t> memhosp.com (chiggerson@memhosp.com) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Re: Cryostats Message-ID: I am also in the market for a new cryostat and would like to hear from USERS. Is there anyone out there with any of the newer cryostats from Sakura, Leica, Microm, or others? I have already talked to my sales reps ---- I want to hear from labs who have used the instruments. Thanks, Cindy Cindy Higgerson HTL(ASCP) Surgical Pathology Supervisor Memorial Hospital Belleville, Illinois From jcline <@t> wchsys.org Thu Sep 9 12:44:24 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Annette, synaptophysin Message-ID: <000501c49694$a5dff2a0$1d2a14ac@wchsys.org> I use a steamer (Black & Decker) for retrieval. I put the slides into Cell Marque's Trilogy and steam for 20 min. Take the slides out of the steamer and let cool for 10 min. Place slides into APK wash (ventana) for 5 min. I incubate for 32 min. No amplification or blocking. I have no problem with light staining. ***** 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 JQB7 <@t> CDC.GOV Thu Sep 9 13:04:58 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] malarial pigment removal Message-ID: Histonetters: What is the simplest and most frequently used procedure for removal of malarial and/or formalin pigment? I have heard concentrated sulfuric acid works well but I do not know the time necessary in the acid. I have used saturated alcoholic picric acid in the past but no longer have picric acid in my lab. I have also heard of the Gridley technique. I just wondered what everyone else is using. Thanks in advance! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 From arvind <@t> nbrc.res.in Thu Sep 9 13:09:23 2004 From: arvind <@t> nbrc.res.in (Arvind Pundir) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] problem with PCN Message-ID: <7DE0291A47C4BC4D9CDEF65D5017CEF0A6D6@mail.nbrc.res.in> hi u can try using TBS with pH 9.0 under microwave oven at full power (450W) for antigen reteriving i have found it useful to some extent , further u should increase the time for blocking serum upto 1 hr hope it works well for arvind@nbrc.res.in From Barry.R.Rittman <@t> uth.tmc.edu Thu Sep 9 13:43:13 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] quantitative graded scale for IHC Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0F131EB@UTHEVS3.mail.uthouston.edu> I would agree with the comments below from Greg. The major problem with comparing stain intensities between sections is that you generally have no real measure of the section thickness for an individual section. If you are cutting at 5 microns there can easily be a one micron difference between sections, which translated into differences in volume of tissue and intensity of staining can be significant. One way around this is to prepare a block of tissue that is embedded with the sample. The block can be of a homogeneous protein material stained for example with a Procion dye. You can, if you have no social life, prepare sections of this material, dissect out a measured area with a blade (using a dissecting scope) and weigh it. You can then relate this volume of tissue to its intensity of staining and work back to relate this to section thickness. If a cylinder of this material is embedded with the block it provides a built in control that will allow comparison between sections. his does not of course take into account any differences in staining due to differences in staining conditions for different slides. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Greg Dobbin Sent: Thursday, September 09, 2004 7:54 AM To: Carla M Conway Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] quantitative graded scale for IHC Hi Carla, I certainly may be corrected by others on this but I personally don't think you will obtain statistically valid data by grading intensity of the reaction between cells or fields or sections. There is just so much room for subjectivity and for that matter, variability between runs or perhaps even between slides within runs! What you may want to try is calculating a ratio of positive to negative (perhaps pos. cells vs. neg. cells in a given area). For instance: place a grid with 50 or 100 intersects on the computer monitor, choose a microscopic field (in a random or systematic- random manner), and then at each intersect decide whether the reaction at that point is positive or negative. If at any particular intersect you have trouble to decide whether it is pos or neg, have a system that does not permit subjectivity, such as: for such points where the call could go either way, look at the space imediately to the left of the y-axis and above the x-axis and then make your decision. Determination of the number of fields you need to sample in order to obtain statistically valid data is very important and a matter for a statistician to explain and/or determine for you (which I most definately am not) unless you happen to have that training yourself. Another idea would be to have the image analysis software either measure the area of or count the number of pixels in a given area (field) that are "this red or redder". This involves choosing a relatively arbitrational threshold for the degree of redness that is to be measured or counted and applying the exact same setting to all other fields and sections in the study. My personal experience (using Bioquant NOVA) has been this sounds great in theory but was very difficult to pull off! The good old-fashioned counting was much more reliable and for that matter, easier to defend in presentation or publication. I am eager to hear other ideas or methods that might also work! Good luck. Greg To: Histonet@lists.utsouthwestern.edu From: "Carla M Conway" Date sent: Thu, 9 Sep 2004 07:22:02 -0700 Copies to: Subject: [Histonet] quantitative graded scale for IHC > Hello, > > I would like to devise a quantitative graded scale for IHC results which > would provide more info than just +/- staining. > I have a reference where IHC staining intensity was rated (from pale pink > to dark red) using alkaline phosphatase/Vector-Red, however we are using > Envision+/AEC which seems to be an all or nothing (red or no red) chromogen > with no gradation. We just received ImagePro image analysis software, so > this may be another route to try. Thanks in advance for any comments or > suggestions you may have. > > Sincerely, > > Carla Conway > Western Fisheries Research Center > Seattle, WA > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 Jackie.O'Connor <@t> abbott.com Thu Sep 9 14:12:22 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] quantitative graded scale for IHC Message-ID: When it was my job to perform quality assurance testing of ER/PgR IHC kits, I used a 'validated' slide (actually, a photomicrograph) with well documented ER or PgR positive cells, stained with a master kit. Various positive cells were labeled as +4, +3, +2, +1, and +0 (negative) -yes, very subjective, but two people had to agree on the positive cells. In the process of evaluating the quality of a new kit, or testing the stability of an on-market kit - I used the kit to stain known positive tumors. I counted each +4 through =0 cell within a pre-determined grid area to ensure the kit on test was giving appropriate results by conforming to the quality assurance guidelines pre-established for that test. (similar to what Greg has stated below) I'm sure that with the advent of image analysis, my job would have been much simpler and much less subjective. Thank Goodness they killed that product line, and I was able to go on to more interesting work. I'm curious as to how current antibody manufacturers perform stabilty testing on their products. Jackie O' "Barry R Rittman" Sent by: histonet-bounces@lists.utsouthwestern.edu 09/09/2004 01:43 PM To: "histonet" cc: Subject: RE: [Histonet] quantitative graded scale for IHC I would agree with the comments below from Greg. The major problem with comparing stain intensities between sections is that you generally have no real measure of the section thickness for an individual section. If you are cutting at 5 microns there can easily be a one micron difference between sections, which translated into differences in volume of tissue and intensity of staining can be significant. One way around this is to prepare a block of tissue that is embedded with the sample. The block can be of a homogeneous protein material stained for example with a Procion dye. You can, if you have no social life, prepare sections of this material, dissect out a measured area with a blade (using a dissecting scope) and weigh it. You can then relate this volume of tissue to its intensity of staining and work back to relate this to section thickness. If a cylinder of this material is embedded with the block it provides a built in control that will allow comparison between sections. his does not of course take into account any differences in staining due to differences in staining conditions for different slides. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Greg Dobbin Sent: Thursday, September 09, 2004 7:54 AM To: Carla M Conway Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] quantitative graded scale for IHC Hi Carla, I certainly may be corrected by others on this but I personally don't think you will obtain statistically valid data by grading intensity of the reaction between cells or fields or sections. There is just so much room for subjectivity and for that matter, variability between runs or perhaps even between slides within runs! What you may want to try is calculating a ratio of positive to negative (perhaps pos. cells vs. neg. cells in a given area). For instance: place a grid with 50 or 100 intersects on the computer monitor, choose a microscopic field (in a random or systematic- random manner), and then at each intersect decide whether the reaction at that point is positive or negative. If at any particular intersect you have trouble to decide whether it is pos or neg, have a system that does not permit subjectivity, such as: for such points where the call could go either way, look at the space imediately to the left of the y-axis and above the x-axis and then make your decision. Determination of the number of fields you need to sample in order to obtain statistically valid data is very important and a matter for a statistician to explain and/or determine for you (which I most definately am not) unless you happen to have that training yourself. Another idea would be to have the image analysis software either measure the area of or count the number of pixels in a given area (field) that are "this red or redder". This involves choosing a relatively arbitrational threshold for the degree of redness that is to be measured or counted and applying the exact same setting to all other fields and sections in the study. My personal experience (using Bioquant NOVA) has been this sounds great in theory but was very difficult to pull off! The good old-fashioned counting was much more reliable and for that matter, easier to defend in presentation or publication. I am eager to hear other ideas or methods that might also work! Good luck. Greg To: Histonet@lists.utsouthwestern.edu From: "Carla M Conway" Date sent: Thu, 9 Sep 2004 07:22:02 -0700 Copies to: Subject: [Histonet] quantitative graded scale for IHC > Hello, > > I would like to devise a quantitative graded scale for IHC results which > would provide more info than just +/- staining. > I have a reference where IHC staining intensity was rated (from pale pink > to dark red) using alkaline phosphatase/Vector-Red, however we are using > Envision+/AEC which seems to be an all or nothing (red or no red) chromogen > with no gradation. We just received ImagePro image analysis software, so > this may be another route to try. Thanks in advance for any comments or > suggestions you may have. > > Sincerely, > > Carla Conway > Western Fisheries Research Center > Seattle, WA > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kim.snyder <@t> emphysis-med.com Thu Sep 9 15:20:30 2004 From: kim.snyder <@t> emphysis-med.com (Kim Snyder) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Alcian Blue Message-ID: > What CPT code should I use for the coding of Alcian Blue? > > Thank you for your help > > Kim Snyder > EmPhysis Medical Management From Rcartun <@t> harthosp.org Thu Sep 9 15:36:25 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] sSILVER STAIN FOR CAT SCRATCH FEVER Message-ID: Cat scratch disease; not cat scratch fever (a popular song in the 80's - I think). We use immunoperoxidase staining with a monoclonal antibody (clone H2A10) from Novus Biologicals. 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 >>> Diana McCaig 09/02/04 11:58AM >>> Hi I was wondering if anyone could suggest what is the recommended special stain to demonstrate Cat Scratch Fever (coccibacilli) in a lymph node? Diana McCaig, R.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bucana <@t> audumla.mdacc.tmc.edu Thu Sep 9 16:15:23 2004 From: bucana <@t> audumla.mdacc.tmc.edu (Corazon D. Bucana) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] benzyl benzoate clearing agent Message-ID: <5.1.1.6.0.20040909160428.00a97cc0@audumla.mdacc.tmc.edu> I would appreciate any information on the use of benzyl alcohol/benzyl benzoate as a clearing agent for tissues. I understand this could be used in combination with fluorescence staining to image thick sections but I have not seen any reference to it in the literature (Pub Med search). I hope somebody out there can direct me to a book or journal that mentions this agent. Thanks, Cora Bucana From Barry.R.Rittman <@t> uth.tmc.edu Thu Sep 9 16:49:08 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] benzyl benzoate clearing agent Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0F13210@UTHEVS3.mail.uthouston.edu> Benzyl benzoate (phenyl methyl benzoate) Refractive index 1.5685 to 1.570. Miscible with alcohol, chloroform, ether and xylene. Never heard of it being used for fluorescence microscopy. It has been used as a clearing agent and mountant for thin museum specimens, often with small amount of methyl salicylate added. The problem with benzyl benzoate is that it is intolerant of water so cannot breathe on the specimen while it is being mounted. Hope this helps Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Corazon D. Bucana Sent: Thursday, September 09, 2004 4:15 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] benzyl benzoate clearing agent I would appreciate any information on the use of benzyl alcohol/benzyl benzoate as a clearing agent for tissues. I understand this could be used in combination with fluorescence staining to image thick sections but I have not seen any reference to it in the literature (Pub Med search). I hope somebody out there can direct me to a book or journal that mentions this agent. Thanks, Cora Bucana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mariac <@t> creighton.edu Thu Sep 9 16:51:27 2004 From: mariac <@t> creighton.edu (Maria Christensen) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Tissue transport from processor to embedding station Message-ID: Our research lab is doing histology on a limited budget, so we use reconditioned equipment to keep our costs down. We recently "moved up" to a VIP 2000, but use an old Tissue Tek II embedding center. My question is, how do I keep the tissue warm between the processor and the embedding station? I have the option of purchasing a paraffin pot (from an old Autotechnicon) and using that filled with paraffin to transport & keep the cassettes warm until the tissue is embedded. Are there any other suggestions? (My budget for this is approximately $500-1000, which makes the $200 pot a real consideration!!) Maria Maria A. Christensen Technical Associate Medical Microbiology & Immunology Creighton University School of Medicine 2500 California Plaza Omaha, NE 68178-0404 phone (402) 280-2678 FAX (402) 280-1875 email mariac@creighton.edu From portera203 <@t> yahoo.com Thu Sep 9 17:52:09 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Tissue transport from processor to embedding station In-Reply-To: Message-ID: <20040909225209.69753.qmail@web40906.mail.yahoo.com> Maria - we also have a Tissue Tek II embedding station, however we have the holding reservoir that goes along with it - so we have the 3 separate sections for this instrument. You might want to check into purchasing a used/reconditioned unit, the baskets from the VIP 2000 fit right into it. It also has a chamber for warming your embedding molds if you are using stainless steel molds like we still are. Good Luck! Maria Christensen wrote:Our research lab is doing histology on a limited budget, so we use reconditioned equipment to keep our costs down. We recently "moved up" to a VIP 2000, but use an old Tissue Tek II embedding center. My question is, how do I keep the tissue warm between the processor and the embedding station? I have the option of purchasing a paraffin pot (from an old Autotechnicon) and using that filled with paraffin to transport & keep the cassettes warm until the tissue is embedded. Are there any other suggestions? (My budget for this is approximately $500-1000, which makes the $200 pot a real consideration!!) Maria Maria A. Christensen Technical Associate Medical Microbiology & Immunology Creighton University School of Medicine 2500 California Plaza Omaha, NE 68178-0404 phone (402) 280-2678 FAX (402) 280-1875 email mariac@creighton.edu _______________________________________________ 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!? Shop for Back-to-School deals on Yahoo! Shopping. From portera203 <@t> yahoo.com Thu Sep 9 17:59:06 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Immuno question about CK markers Message-ID: <20040909225906.64544.qmail@web40905.mail.yahoo.com> Need some help everyone. I am taking the QIHC exam and am looking for a little advice about the grading system with regards to Cytokeratin 7 and Cytokeratin 20. While I realize that they would like to have them mirror each other, will they down grade your score if both of your tumors are not completely negative for the opposite marker. I have excellent staining in both tissues that are supposed to be positive for each marker and then foci's of sporadic staining (maybe 2 or 3 spots) that demonstrates positive reaction where the tumor is negative. I am guessing that this is just the nature of the cytokeratin world, that not all tissue components will be negative. Hope this makes sense and someone can give me some idea from their own experience. Thanks in advance for all the help i know i will get. 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!? Express yourself with Y! Messenger! Free. Download now. From rentonlf <@t> bru.wits.ac.za Fri Sep 10 02:34:01 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] re:Tissue transport from processor to embedding longish reply Message-ID: <1094801641.948e1b40rentonlf@bru.wits.ac.za> Maria, Is there a specific reason why you wish to keep your tissue immersed between processor & embedding centre (USA regs or long distance between the 2)? The tissue will come to no harm if allowed to drain before carrying them, in their basket, on a metal or plastic tray to the embedding centre. I don't mean let them get ice cold, and all grungy, but this is how we used to do them in the olden days. In fact, we would just carry the basket on a wad of paper towel to contain the drips. Being clumsy by nature, this is a safer bet for folks like me, rather than carrying a pot of molten wax around! Once there, the cassettes were stood up on the drainage area of the warming plate (these were the old stainless steel & black paint Tissue Teks) and embedded. I remember that the plate could hold the contents of the basket from a 1 liter dip & dunk machine quite comfortably. Ahhh, enough of the reminiscences. Processed tissues will withstand a short stay in an incubator/oven set just above the melting point of your wax, either drained or in a metal pot of wax. FYI, we used to use stainless steel water jugs for this - have a handy spout to pour from. I never found out if they were specifically bought or "relocated" from the canteen. Have a great weekend -----Original Message----- From: Maria Christensen To: histonet@lists.utsouthwestern.edu Date: Thu, 9 Sep 2004 16:51:27 -0500 Subject: [Histonet] Tissue transport from processor to embedding station Our research lab is doing histology on a limited budget, so we use reconditioned equipment to keep our costs down. We recently "moved up" to a VIP 2000, but use an old Tissue Tek II embedding center. My question is, how do I keep the tissue warm between the processor and the embedding station? I have the option of purchasing a paraffin pot (from an old Autotechnicon) and using that filled with paraffin to transport & keep the cassettes warm until the tissue is embedded. Are there any other suggestions? (My budget for this is approximately $500-1000, which makes the $200 pot a real consideration!!) Maria Maria A. Christensen Technical Associate Medical Microbiology & Immunology Creighton University School of Medicine 2500 California Plaza Omaha, NE 68178-0404 phone (402) 280-2678 FAX (402) 280-1875 email mariac@creighton.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From lpwenk <@t> sbcglobal.net Fri Sep 10 03:42:42 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] malarial pigment removal References: Message-ID: <005f01c49712$23eda980$ec25d445@domainnotset.invalid> I've never tried the following on malaria pigment, but it does work on formalin pigment. H&E will look a little different - nuclei slightly paler but still good chromatin material, and only 2 shades of eosin instead of 3. Really only notice the difference if you compare the 2 slides side by side - one with removal and one without. 100 mL of 70% alcohol 3 mL of conc. ammonium hydroxide Mix together in hood just before use. Place deparaffinized and hydrated slides in this mixture. Depending on the amount of formalin pigment, leave slides in for 30 minutes - 3 hours. (I find 1 hour works for most.) Rinse well in water. Stain as usual. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Bartlett, Jeanine" To: Sent: Thursday, September 09, 2004 2:04 PM Subject: [Histonet] malarial pigment removal Histonetters: What is the simplest and most frequently used procedure for removal of malarial and/or formalin pigment? I have heard concentrated sulfuric acid works well but I do not know the time necessary in the acid. I have used saturated alcoholic picric acid in the past but no longer have picric acid in my lab. I have also heard of the Gridley technique. I just wondered what everyone else is using. Thanks in advance! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From barbara.bublava <@t> meduniwien.ac.at Fri Sep 10 06:33:01 2004 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] malarial pigment removal References: Message-ID: <004b01c49729$edc3ec30$1201a8c0@GERICHTS9XOZZ8> I use 5% ammonium hydroxide in tapwater for 20 minutes. At first I used 70% Ethanol, it also did work but later I learned that tapwater is also OK It definetifely works on formalinpigment and according to literature it should also remove malariapigment. We do several stains (HE, Goldner (Tricrome), Elastica, Prussian blue, Van Gieson, Casons(SFOG), Ziehl nieson, Gram, Kluver Barrera ...) and saw no changes, but we never tryed IHC or silverstains! Be shure to rinse well. remains of Ammonia can change the pH of dyingsolutions (for example eosin) so they would get weaker and weaker have a nice weekend Barbara, Vienna ----- Original Message ----- From: "Bartlett, Jeanine" To: Sent: Thursday, September 09, 2004 8:04 PM Subject: [Histonet] malarial pigment removal Histonetters: What is the simplest and most frequently used procedure for removal of malarial and/or formalin pigment? I have heard concentrated sulfuric acid works well but I do not know the time necessary in the acid. I have used saturated alcoholic picric acid in the past but no longer have picric acid in my lab. I have also heard of the Gridley technique. I just wondered what everyone else is using. Thanks in advance! Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bernaweston <@t> hotmail.com Fri Sep 10 07:12:27 2004 From: bernaweston <@t> hotmail.com (Bernadette Weston) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] =?iso-8859-1?q?Fw=3A_Forget_Bush_and_Kerry_-_-_The_M?= =?iso-8859-1?q?=26M=27s_=AE_Brand_Characters_need_your_vote=2E?= Message-ID: >From: "Barbara" >To: ,,"Don d" >,,"Carlozzi Sharen" >,"Bob and Donna Brennan" >,"Bernadette Weston" ,"BARBARA >DUDA" , >Subject: Fw: Forget Bush and Kerry - - The M&M's ® Brand Characters need >your vote. >Date: Fri, 10 Sep 2004 07:04:51 -0400 > >thought that you might find this amusing > >Barbara >----- Original Message ----- >From: Barbara >To: Barbara >Sent: Friday, September 10, 2004 7:00 AM >Subject: Fw: Forget Bush and Kerry - - The M&M's ® Brand Characters need >your vote. > > > >----- Original Message ----- >From: mark.chimel@effem.com >To: barbara.chimel@comcast.net >Sent: Wednesday, September 08, 2004 8:41 AM >Subject: Forget Bush and Kerry - - The M&M's ® Brand Characters need your >vote. > > > > >Mark Chimel >Sr. Process Development Engineer >Masterfoods, Inc. >800 High St. >Hackettstown, NJ 07840 >phone 908-850-2835 >fax 908-850-2697 >mobile 908-619-2903 >email mark.chimel@effem.com > >NOTICE OF CONFIDENTIALITY -- THIS DOCUMENT AND THE INFORMATION CONTAINED IN >IT ARE CONFIDENTIAL AND ARE THE PROPERTY OF MASTERFOODS. THEY MAY NOT IN >ANY WAY BE DISCLOSED, COPIED OR USED BY ANYONE EXCEPT AS EXPRESSLY >AUTHORIZED BY MASTERFOODS. THE DOCUMENT SHOULD ALWAYS BE KEPT IN A SECURE >PLACE AND SHOULD BE DESTROYED OR RETURNED TO MASTERFOODS WHEN IT IS NO >LONGER NEEDED. >----- Forwarded by Mark J. Chimel/HKT/Effem on 09/08/2004 08:40 AM ----- > > Jackie A. Hague > HKT - Effem > 09/02/2004 03:19 PM > > > To: HKT NO Associates List, HKT NO Associates List >A-B@Effem_Americas, HKT NO Associates List C-D@Effem_Americas, HKT NO >Associates List E-H@Effem_Americas, HKT NO Associates List >I-L@Effem_Americas, HKT NO Associates List M-O@Effem_Americas, HKT NO >Associates List P-R@Effem_Americas, HKT NO Associates List >S-U@Effem_Americas, HKT NO Associates List V-Z@Effem_Americas, HKP - Staff >Members, HKP E-mail Users, HKP E-mail Users (A-D)@Effem_Americas, HKP >E-mail Users (E-J)@Effem_Americas, HKP E-mail Users (K-O)@Effem_Americas, >HKP E-mail Users (P-S)@Effem_Americas, HKP E-mail Users >(T-Z)@Effem_Americas, KKV National Office, KKV National Office Managers, >KKV National Office (A-E)@Effem_Americas, KKV National Office >(F-L)@Effem_Americas, KKV National Office (M-R)@Effem_Americas, KKV >National Office (S-Z)@Effem_Americas, KKV Plant, KKV Plant >(A-F)@Effem_Americas, KKV Plant (G-L)@Effem_Americas, KKV Plant >(M-S)@Effem_Americas, KKV Plant (T-Z)@Effem_Americas, CLV ALL Associates > cc: > > > Subject: Forget Bush and Kerry - - The M&M's® Brand >Characters need your vote. > > > > >Hello all - - Your support is needed to enter the M&M's® Brand Characters >into the Madison Avenue Walk of Fame. Our associates can rally around our >favorite characters to improve their celebrity status and affirm their >place in history. The M&M's® Brand Characters are in a race to win >Advertising Week's "America's Favorite Ad Icons" contest -- But they need >your help! > >Here's how you can help: > > a.. Go to http://www1.mms.com/us/ > b.. You'll see the voting button on the right corner of the Home Page > c.. Click "Vote Now" and start with the "Favorite Icon Poll" > d.. Now you can vote to help Red® and friends beat everyone else! > e.. Submit your vote > f.. Next, go to the "Favorite Slogan Poll" and find our legendary "Melts >in your mouth, not in your hands" slogan > g.. Submit your vote > h.. Give yourself a pat on the back for helping our favorite characters! > i.. Or just use the attachments below. > > Please hurry, voting ends September 17th! Thanks for your support. > Jackie Hague > "M&M's" Brand Manager > Masterfoods USA > A MARS Incorporated Company > Hackettstown, NJ > 07840 USA > 908-850-7891 > > > Dear M&M'S® Brand Fan, > Our beloved Spokescandies, Red & Yellow, are in the running for >America's Favorite Advertising Mascot. The top five Mascots, voted on by >fans like you, will be the very first honored on the "Madison Avenue >Advertising Walk of Fame" in New York City. Join the fun and cast your vote >for your favorite mascot! > > It's easy to vote. Just visit the Advertising Week 2004 website. >And while you're there, be sure to vote for your favorite advertising >slogan of all time, like Red's favorite "Melts in your mouth, not in your >hands". And don't forget to tell your friends to vote for their favorite >too! > > Have fun! > The M&M'S® Brand > > ©Mars, Incorporated 2004. All Rights Reserved. > We respect our visitors' privacy. And now, a word from our >lawyers... > Masterfoods USA 800 High Street Hackettstown, NJ 07840 USA > To unsubscribe, please click here. > > > > _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar – get it now! http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/ From JNocito <@t> Pathreflab.com Fri Sep 10 08:01:59 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Alcian Blue In-Reply-To: Message-ID: We use 88313 when we are staining for mucopolysaccarhides. 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 Kim Snyder Sent: Thursday, September 09, 2004 3:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Alcian Blue > What CPT code should I use for the coding of Alcian Blue? > > Thank you for your help > > Kim Snyder > EmPhysis Medical Management _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Fri Sep 10 09:00:03 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] synaptophysin Message-ID: Hello Annette. We use Ventana's lyoph. Antibody with standard CC1 retrieval and primary on for 24 min. No amplification required. Good luck. -----Original Message----- From: JAnorthernexp@cs.com [mailto:JAnorthernexp@cs.com] Sent: Wednesday, September 08, 2004 7:07 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] synaptophysin anyone using a ventana stainer and antibodies have a retrieval method using a decloker? Ours stains very light. Antibody is on for 32 mins we did have it set at 16mins. But increasing the time did not prove to make much difference. Any other ideas?? The tissue is formalin fixed and paraffin embedded. Thanks Annette _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Fri Sep 10 09:07:54 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Re: Cryostats Message-ID: I would like to tell you about my cryostat, but I think I see vultures circling over my head. BIG BROTHER IS WATCHING!!! Good luck in your search. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 -----Original Message----- From: chiggerson@memhosp.com [mailto:chiggerson@memhosp.com] Sent: Thursday, September 09, 2004 11:53 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Cryostats I am also in the market for a new cryostat and would like to hear from USERS. Is there anyone out there with any of the newer cryostats from Sakura, Leica, Microm, or others? I have already talked to my sales reps ---- I want to hear from labs who have used the instruments. Thanks, Cindy Cindy Higgerson HTL(ASCP) Surgical Pathology Supervisor Memorial Hospital Belleville, Illinois _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JGordon <@t> cellmarque.com Fri Sep 10 09:16:06 2004 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Immuno question about CK markers Message-ID: Amy, one way that we have been able to assist people with those issues is to check on the pretreatment that is being used. In the past, we have spoken to people using CK20 that said that they had received staining on breast tissue with their antibody. They were pretreating it with enzyme digestion. We suggested trying HIER with our Trilogy EDTA and it gave them the same specific staining in the GI tissue that they were receiving originally, but also eliminated the false positives that they were receiving in the breast tissue with the CK20. Perhaps you should try an alternate epitope retrieval technique to see if that eliminates your sporadic false positive staining. This may not apply to you, but it is just a suggestion to help you out in case the pretreatment is relevant to your tests. If you have any further questions, let me know. Jeff Gordon Cell Marque Corp. 1-800-665-7284, Ext. 12 http://www.cellmarque.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Porter Sent: Thursday, September 09, 2004 4:59 PM To: Histonet Subject: [Histonet] Immuno question about CK markers Need some help everyone. I am taking the QIHC exam and am looking for a little advice about the grading system with regards to Cytokeratin 7 and Cytokeratin 20. While I realize that they would like to have them mirror each other, will they down grade your score if both of your tumors are not completely negative for the opposite marker. I have excellent staining in both tissues that are supposed to be positive for each marker and then foci's of sporadic staining (maybe 2 or 3 spots) that demonstrates positive reaction where the tumor is negative. I am guessing that this is just the nature of the cytokeratin world, that not all tissue components will be negative. Hope this makes sense and someone can give me some idea from their own experience. Thanks in advance for all the help i know i will get. 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!? Express yourself with Y! Messenger! Free. Download now. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ASelf <@t> gmhsc.com Fri Sep 10 09:25:51 2004 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Leica 1800 cryostat manual Message-ID: <39836CD6DB61654E8F95A35898C921860A87ED@exchange.gmhpost.com> Does anyone have a manual for the old Leica 1800 cryostat that they could share with me or don't need anymore? We (biomed dept.) has misplaced ours. Thanks, amy NOTE: The information contained in this message may be privileged, 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 this message and deleting it from your computer. Thank you. From padunnje <@t> iupui.edu Fri Sep 10 09:48:52 2004 From: padunnje <@t> iupui.edu (Dunn-Jena, Patsy A) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Question regarding 10% Neutral buffered formalin/Immuno Message-ID: I have a graduate student who has come to me with a question I cannot answer as I have limited Immunohistochemical experience. One of her advisers wants her to go to using an EDTA decalcification recipe which contains the 10% NBF in it which is a recipe we used on all of his previous work. She asked me about the NBF crosslinking her immuno sites as she has been using an EDTA decalcification only recipe. Some input here would be nice so I can better explain it to her. Patsy A. Dunn-Jena, RVT, LAT, HT (ASCP) Mineralized Tissue and Histology Research Laboratory Indiana University School of Dentistry 1121 W. Michigan Street, Room 238 Indianapolis, IN 46202 padunnje@iupui.edu (317)274-0544 From bucana <@t> audumla.mdacc.tmc.edu Fri Sep 10 09:49:58 2004 From: bucana <@t> audumla.mdacc.tmc.edu (Corazon D. Bucana) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] benzyl benzoate clearing agent In-Reply-To: <566FB0B522443D43AF02D2ADBE35A6F0F13210@UTHEVS3.mail.uthous ton.edu> Message-ID: <5.1.1.6.0.20040910093746.00a8f800@audumla.mdacc.tmc.edu> Thank you for the info. I heard a talk about confocal microscopy of embryos immunolabeled for some protein that I can't remember now but the speaker mentioned benzyl benzoate as a clearing agent after the embryos were immunolabeled and claims that they are able to create a stack of optical sections to a depth of 300 microns . This is quite a feat because most often we only look at 50-100 microns thick sections. I wrote the speaker asking for the procedure but so far I haven't had a response which is why I thought somebody at the Histonet will have an idea about this agent. Thanks Cora Bucana >Benzyl benzoate (phenyl methyl benzoate) >Refractive index 1.5685 to 1.570. >Miscible with alcohol, chloroform, ether and xylene. >Never heard of it being used for fluorescence microscopy. >It has been used as a clearing agent and mountant for thin museum >specimens, often with small amount of methyl salicylate added. The >problem with benzyl benzoate is that it is intolerant of water so cannot >breathe on the specimen while it is being mounted. >Hope this helps >Barry > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Corazon >D. Bucana >Sent: Thursday, September 09, 2004 4:15 PM >To: Histonet@lists.utsouthwestern.edu >Subject: [Histonet] benzyl benzoate clearing agent > >I would appreciate any information on the use of benzyl alcohol/benzyl >benzoate as a clearing agent for tissues. I understand this could be >used >in combination with fluorescence staining to image thick sections but I >have not seen any reference to it in the literature (Pub Med search). I > >hope somebody out there can direct me to a book or journal that mentions > >this agent. > >Thanks, > >Cora Bucana > > >_______________________________________________ >Histonet mailing list >Histonet@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 mariac <@t> creighton.edu Fri Sep 10 10:04:40 2004 From: mariac <@t> creighton.edu (Maria Christensen) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Tissue transport from processor to embedding station Message-ID: Thanks to all who replied. To answer some of your questions, our processor and embedding center are in separate rooms now, but not far from each other (10-12 steps). Our concern was keeping the tissue warm so that we could embed quickly when needed without having to warm the tissue up. We thought of just transferring in a beaker of melted paraffin, but sometimes we can't get to all the cassettes right away, so the wax would begin to solidify. Unfortunately we don't have an oven in the lab where the embedding equipment is. Our Tissue Tek II embedding station has a heated compartment for the molds, but no compartment for the cassettes. The manual mentions an infiltration process carrier, and a vacuum infiltrator, but these accessories did not come with our reconditioned equipment. I would like to know more about the holding reservoir that Amy Porter mentioned - part no., where purchased, etc. because if the basket from the VIP fits in it, that would be ideal. Again, thanks for your ideas- I'm a firm believer in not reinventing the wheel! Maria Maria A. Christensen Technical Associate Medical Microbiology & Immunology Creighton University School of Medicine 2500 California Plaza Omaha, NE 68178-0404 phone (402) 280-2678 FAX (402) 280-1875 email mariac@creighton.edu From padunnje <@t> iupui.edu Fri Sep 10 10:08:57 2004 From: padunnje <@t> iupui.edu (Dunn-Jena, Patsy A) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Question regarding 10% Neutral buffered formalin/Immuno Message-ID: Additional information. The tissue is the palate/maxillary teeth of mice. It is fixed in 10% NBF for 24 hours prior to starting to decal in a 0.25 M EDTA which takes about 4 weeks. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Dunn-Jena, Patsy A Sent: Friday, September 10, 2004 9:49 AM To: Histonet Subject: [Histonet] Question regarding 10% Neutral buffered formalin/Immuno I have a graduate student who has come to me with a question I cannot answer as I have limited Immunohistochemical experience. One of her advisers wants her to go to using an EDTA decalcification recipe which contains the 10% NBF in it which is a recipe we used on all of his previous work. She asked me about the NBF crosslinking her immuno sites as she has been using an EDTA decalcification only recipe. Some input here would be nice so I can better explain it to her. Patsy A. Dunn-Jena, RVT, LAT, HT (ASCP) Mineralized Tissue and Histology Research Laboratory Indiana University School of Dentistry 1121 W. Michigan Street, Room 238 Indianapolis, IN 46202 padunnje@iupui.edu (317)274-0544 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bwhitaker <@t> brownpathology.com Fri Sep 10 10:31:21 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] IHC billing/payment question Message-ID: <000001c4974b$395af460$3601a8c0@brownpathology.net> Hi All, A question recently came up that I'd like to ask about. We are a new lab, and are not yet doing IHC. We are planning to do so in the near future, but one of our people recently heard that Medicare was not paying for IHC, in some instances, unless there were some particular ICD-9 codes associated with it. I did find something referring to H. Pylori testing, and there was a group of ICD-9 codes that were acceptible in this instance and all others were not. Does anyone know of a definitive listing of markers and the codes that must be associated? I've never had to be involved in this end of things before, and I feel totally clueless! I don't want to push to start doing IHC if reimbursement is becoming a serious problem. HELP! Thanks, Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 From John.Auld <@t> whnt.nhs.uk Fri Sep 10 10:53:52 2004 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Re: Question regarding 10% Neutral buffered Message-ID: Patsy It shouldn't make any difference to most immuno stains. The NBF in the EDTA will continue to act as a fixative and may, just may overfix some Ags, some will be OK. I would suspect that 24 hrs primary fixation is insufficient prior to decalcification anyway, so the use of NBF in EDTA maybe beneficial. However, if some results have been obtained prior to changing the protocol it maybe better not to change. Regards John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral Internal extn 2560 External Tel 0151 604 7025 Message: 25 Date: Fri, 10 Sep 2004 10:08:57 -0500 From: "Dunn-Jena, Patsy A" Subject: RE: [Histonet] Question regarding 10% Neutral buffered formalin/Immuno To: "Histonet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Additional information. The tissue is the palate/maxillary teeth of mice. It is fixed in 10% NBF for 24 hours prior to starting to decal in a 0.25 M EDTA which takes about 4 weeks. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Dunn-Jena, Patsy A Sent: Friday, September 10, 2004 9:49 AM To: Histonet Subject: [Histonet] Question regarding 10% Neutral buffered formalin/Immuno I have a graduate student who has come to me with a question I cannot answer as I have limited Immunohistochemical experience. One of her advisers wants her to go to using an EDTA decalcification recipe which contains the 10% NBF in it which is a recipe we used on all of his previous work. She asked me about the NBF crosslinking her immuno sites as she has been using an EDTA decalcification only recipe. Some input here would be nice so I can better explain it to her. Patsy A. Dunn-Jena, RVT, LAT, HT (ASCP) Mineralized Tissue and Histology Research Laboratory Indiana University School of Dentistry 1121 W. Michigan Street, Room 238 Indianapolis, IN 46202 padunnje@iupui.edu (317)274-0544 From TJJ <@t> Stowers-Institute.org Fri Sep 10 11:24:38 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Re: benzyl benzoate clearing agent Message-ID: Corazon, I am marginally familiar with using BABB (mixture of benzyl alcohol/benzyl benzoate) as a clearing agent for whole mount embryos, and a quick google search for this yielded this website which uses it in Xenopus for confocal: http://froglab.biology.utah.edu/Tools_and_techniques/body_tools_and_tech niques.html. There is a link for using it in clearing, and one for using it for mounting. Hope this helps! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From jchandler <@t> ial-fa.com Fri Sep 10 11:28:13 2004 From: jchandler <@t> ial-fa.com (John Chandler) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] benzyl benzoate clearing agent References: <5.1.1.6.0.20040910093746.00a8f800@audumla.mdacc.tmc.edu> Message-ID: <003901c49753$2c3c0ff0$02000000@JohnChandler> That speaker might have been Dave Gard from the Biology Department at the University of Utah. You can check out his lab at: http://froglab.biology.utah.edu/next.html-ssi --John jchandler@ial-fa.com 970-217-1321 ----- Original Message ----- From: "Corazon D. Bucana" To: "Barry R Rittman" Cc: Sent: Friday, September 10, 2004 8:49 AM Subject: RE: [Histonet] benzyl benzoate clearing agent > Thank you for the info. I heard a talk about confocal microscopy of > embryos immunolabeled for some protein that I can't remember now but the > speaker mentioned benzyl benzoate as a clearing agent after the embryos > were immunolabeled and claims that they are able to create a stack of > optical sections to a depth of 300 microns . This is quite a feat because > most often we only look at 50-100 microns thick sections. I wrote the > speaker asking for the procedure but so far I haven't had a response which > is why I thought somebody at the Histonet will have an idea about this agent. > Thanks > Cora Bucana > > > >Benzyl benzoate (phenyl methyl benzoate) > >Refractive index 1.5685 to 1.570. > >Miscible with alcohol, chloroform, ether and xylene. > >Never heard of it being used for fluorescence microscopy. > >It has been used as a clearing agent and mountant for thin museum > >specimens, often with small amount of methyl salicylate added. The > >problem with benzyl benzoate is that it is intolerant of water so cannot > >breathe on the specimen while it is being mounted. > >Hope this helps > >Barry > > > > > >-----Original Message----- > >From: histonet-bounces@lists.utsouthwestern.edu > >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Corazon > >D. Bucana > >Sent: Thursday, September 09, 2004 4:15 PM > >To: Histonet@lists.utsouthwestern.edu > >Subject: [Histonet] benzyl benzoate clearing agent > > > >I would appreciate any information on the use of benzyl alcohol/benzyl > >benzoate as a clearing agent for tissues. I understand this could be > >used > >in combination with fluorescence staining to image thick sections but I > >have not seen any reference to it in the literature (Pub Med search). I > > > >hope somebody out there can direct me to a book or journal that mentions > > > >this agent. > > > >Thanks, > > > >Cora Bucana > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@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 Barry.R.Rittman <@t> uth.tmc.edu Fri Sep 10 12:16:58 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Question regarding 10% Neutral buffered formalin/Immuno Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0F13257@UTHEVS3.mail.uthouston.edu> The original idea of adding formalin or glutaraldehyde to the EDTA was to prevent maceration during the long exposure to EDTA solutions. In my experience these aldehydes significantly decrease the rate of EDTA demineralization. This may well be due to increased cross linking, especially if the original fixation was of relatively short duration. Have no evidence or references for this. My advice would be to use EDTA pH7.2 to 7.4 to eliminate this as a potential problem. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dunn-Jena, Patsy A Sent: Friday, September 10, 2004 9:49 AM To: Histonet Subject: [Histonet] Question regarding 10% Neutral buffered formalin/Immuno I have a graduate student who has come to me with a question I cannot answer as I have limited Immunohistochemical experience. One of her advisers wants her to go to using an EDTA decalcification recipe which contains the 10% NBF in it which is a recipe we used on all of his previous work. She asked me about the NBF crosslinking her immuno sites as she has been using an EDTA decalcification only recipe. Some input here would be nice so I can better explain it to her. Patsy A. Dunn-Jena, RVT, LAT, HT (ASCP) Mineralized Tissue and Histology Research Laboratory Indiana University School of Dentistry 1121 W. Michigan Street, Room 238 Indianapolis, IN 46202 padunnje@iupui.edu (317)274-0544 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From giorgia.setti <@t> kcl.ac.uk Fri Sep 10 12:23:54 2004 From: giorgia.setti <@t> kcl.ac.uk (Giorgia Setti) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] (no subject) Message-ID: <1094837034.4141e32a9fa7a@impmail.kcl.ac.uk> -- Giorgia Setti giorgia.setti@kcl.ac.uk hello histonetters!!!! i am desperate because I can't find a good antibody for MCP-1 staining on rat frozen sections;I tried the MCP-1 antibody of Abcam but it didn't work;do you know any other antibody tested on frozen section?? thank you Giorgia ---------------------------------------------------------------- This message was sent using IMP, the Internet Messaging Program. From Dawn.Olszewski <@t> SGMC.ORG Fri Sep 10 12:29:18 2004 From: Dawn.Olszewski <@t> SGMC.ORG (Olszewski, Dawn) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Nemesis stainer Message-ID: <825F2CDD64ACF34D85E5E5CCC9EA01320557EA2B@email.sgmc.org> Does anyone have any positive or negative comments about the Nemesis IHC stainer? Any info would be appreciated. Thanks in advance. Dawn Olszewski HTL(ASCP)QIHC South Georgia Medical Center From portera203 <@t> yahoo.com Fri Sep 10 12:45:54 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Tissue transport from processor to embedding station In-Reply-To: Message-ID: <20040910174554.28637.qmail@web40901.mail.yahoo.com> Maria - reply to me (off list) if you want and supply me with your fax number and i can fax you pictures of what we have here. It is a Thermal Console we have model #4585 from Miles/Sakura. We also walk several feet to our embedding station and always have just placed our cassettes into the reservoir then let them sit for a few minutes to rewarm. They usually do not get that cold in that short of a time span. Maria Christensen wrote: Thanks to all who replied. To answer some of your questions, our processor and embedding center are in separate rooms now, but not far from each other (10-12 steps). Our concern was keeping the tissue warm so that we could embed quickly when needed without having to warm the tissue up. We thought of just transferring in a beaker of melted paraffin, but sometimes we can't get to all the cassettes right away, so the wax would begin to solidify. Unfortunately we don't have an oven in the lab where the embedding equipment is. Our Tissue Tek II embedding station has a heated compartment for the molds, but no compartment for the cassettes. The manual mentions an infiltration process carrier, and a vacuum infiltrator, but these accessories did not come with our reconditioned equipment. I would like to know more about the holding reservoir that Amy Porter mentioned - part no., where purchased, etc. because if the basket from the VIP fits in it, that would be ideal. Again, thanks for your ideas- I'm a firm believer in not reinventing the wheel! Maria Maria A. Christensen Technical Associate Medical Microbiology & Immunology Creighton University School of Medicine 2500 California Plaza Omaha, NE 68178-0404 phone (402) 280-2678 FAX (402) 280-1875 email mariac@creighton.edu _______________________________________________ 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!? Yahoo! Mail Address AutoComplete - You start. We finish. From gmacke <@t> shrinenet.org Fri Sep 10 13:06:03 2004 From: gmacke <@t> shrinenet.org (Macke, Gail) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Work load for research labs doing GMA Message-ID: Dear Histonetters, Could some one tell me what is considered the average work load for cutting GMA blocks. How many Blocks per year embedded? How many slides cut per year and stained per year? We use Technovit 7100 for processing and embedding. We are a Core Research Lab that also does paraffin and frozen processing and cutting. Most of the information I have found relates to Clinical Labs doing paraffin and frozen work. I haven't found much related to Research Labs. Any information would be helpful. Thank you for taking the time to read and reply to my questions. Gail Macke, HTL Shriners Hospital for Children: Shriners Burns Hospital- Cincinnati Cincinnati, Ohio, USA Gmacke@Shrinenet.org CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From Rhonda.Allen2 <@t> med.va.gov Fri Sep 10 14:30:45 2004 From: Rhonda.Allen2 <@t> med.va.gov (Allen, Rhonda) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] IHC on Frozen Sections Message-ID: <707BC118BC44D3118E3B0000F81F1E7605A393DA@VHAKANEXC1> Hello, I am trying to do the T & B cells on frozen section tonsil for my IHC exam due at the end of this month. I have tried multiple procedures, and have not yet found one that makes me happy, or looks like the one in the picture at the IHC home page. If someone could e-mail me their procedures (mostly from cutting to antibody application) it would be greatly appreciated. By the way, my cryostat will not cut thinner than 4-6 microns. Thanks in advance, Rhonda Allen VA Medical Center From ajaivyas <@t> gmail.com Fri Sep 10 20:12:43 2004 From: ajaivyas <@t> gmail.com (Ajai Vyas) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] GFP in a fixed brain tissue Message-ID: <934be6370409101812725ddc95@mail.gmail.com> Hi, Regards from a new entrants to histonet. I have GFP labelled parasites sitting in brain of my rats. I need to slice the brain and locate these parasites. Will GFP still be present if I use fixatives like formaldehyde or organic solvents like xylene? What fixing protocols are best for preserving GFP signal? Thanks in advance, AJAI From Don.Birgerson <@t> leica-microsystems.com Fri Sep 10 11:20:57 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Leica 1800 cryostat manual Message-ID: Hi Amy, I be happy to mail you a copy of the "User Manual" if you will give me your street address. Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Amy Self" Sent by: To: histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] Leica 1800 cryostat manual 09/10/2004 09:25 AM Does anyone have a manual for the old Leica 1800 cryostat that they could share with me or don't need anymore? We (biomed dept.) has misplaced ours. Thanks, amy NOTE: The information contained in this message may be privileged, 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 this message and deleting it from your computer. Thank you. _______________________________________________ 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 vbaker60 <@t> yahoo.com Sat Sep 11 08:53:05 2004 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Off Topic - question about class action suit against your "employer" Message-ID: <20040911135305.17671.qmail@web50105.mail.yahoo.com> Good morning, This is a question that I hope very few of us on the list-server have ever had to, are currently, or will ever have to deal with/answer personally. A week ago yesterday the not-for-profit/NCI (Cancer Center Support Grant)funded organization I work for gave us WARN notification that told us we would be permanently "layed-off" as of Sept. 10 unless an agreement between our facility and the government was made. As negotiations were being worked for a merger with "other" institutions there might still be a way to work out our difficulties and keep our institution open. The specifics of this merger are unknown and we have had no specific information given to us. As of yesterday afternoon we were informed that we would have to call in on Monday morning to see if we would be allowed access to the building and work. The controversy is this: 1. According to law we are to be notified at least 60 days in advance of a permanent closing. This has not been done. 2. We lose any severance or unused vacation pay still owed to us under a "normal" - I use this term loosely - closing. 3. Insurance benefits - being able to apply for COBRA or get clear information as to what we could do to purchase insurance on our own has been non existent and our HR person left her position as of Friday as well. This is not part of the actual legal proceedings just concerns of all employees. 4. Retirement plans/annuities are a separate issue as not all employees were involved in these benefits. We are a very small institute of less than 150 people in NYS. I'm looking to see if anyone else on the server has had this occur to them and whether or not they did pursue a legal action to get the benefits due to them. I'd also like to know if anyone has information or people/places I could contact about what most have gotten in terms of litigation. The issues I listed are the most pertinent, as with many of these closings other issues revolve around the main ones. Many of these people have been here since the late 70's or early 80's. Yesterday as most of us were cleaning out offices and trying to organize our labs, I encountered so many different human reactions and emotions. When I left my lab last night it was just around sundown and shadows were cast over some of my equipment and benches after I had shut down the lights (I have a wall of windows that I've always loved to watch the seasons or the weather)and it was a very sad moment. Would I ever see it again? When you are layed off or a company closes it's a 'done deal' and you can move on. When you just seem to get a 'stay of termination' that leaves you no where closer to resolution it wears you down. If anyone has anything that they could pass on I would very much appreciate it. Thank you very much. Vikki Baker Institute for Cancer Prevention Valhalla, NY 10595 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From JColCLEFA <@t> aol.com Sat Sep 11 12:31:15 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Decal/EDTA IHC Message-ID: <147.33858726.2e749063@aol.com> Decal/EDTA IHC In my lab, we use 10% neutral buffered formalin as a fixative for virtually all our material. We use a decal solution which contains both formalin and EDTA, and we use an "antigen retrieval" solution which contains EDTA for a significant number of proteins we wish to label. In my experience, certain primary antibodies will not work on material which was decalcified in a formalin/ formic acid/ sodium citrate/EDTA decal solution, but the vast majority will. (Pax 5 is one antibody which won't work for us on decalcified items) My suggestion is to test all your variables before settling on your new EDTA decal. My assumption is that you run the risk of losing antigenicity on a few antibodies. To answer the question-- yes. Formalin will crosslink her antigenic sites, antigen (or epitope) retrieval procedures are successful at "decrosslinking" to enable labelling. My lab stains about 400 IHC slides daily, we formalin fix and subsequently perform antigen retrieval on virtually all our tissue. 40-50% of our work is on bone marrow specimens, and half of those are decalcified biopsies. JC--HT(QIHC) From madaryhtl <@t> juno.com Sat Sep 11 14:03:49 2004 From: madaryhtl <@t> juno.com (madaryhtl@juno.com) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Being laid off Message-ID: <20040911.120350.2741.777302@webmail17.lax.untd.com> My last company went through this twice in the past year. Although they were for profit, some things to consider is if it is an "at will" employment forget the 60 days, cobra is an entitlement based on if you had coverage and is a Federal Law they will have to abide by(plus you can get it yourself), the retirement annuities issues is based on vesting times established at the time of employment, and unused vacation was paid(by law), no sick time or floating days were paid, and severance was not a given for everyone, some folks got nothing. This is wwhat happened at this place, a pharma company. Nick(Rocky) Madary, HT,HTL(ASCP)QIHC Histology Manager, Medimmune Inc Cell-3012334950 Work-3013984745 Fax-3013989745 From wecare <@t> qualityhistology.com Sat Sep 11 14:21:54 2004 From: wecare <@t> qualityhistology.com (wecare@qualityhistology.com) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Being laid off References: <20040911.120350.2741.777302@webmail17.lax.untd.com> Message-ID: <001401c49834$c20d7640$0efea8c0@internetconnect.net> A word of caution about COBRA. If your company is no longer in business, the insurance company will drop you right away also. COBRA applies only to the organizations still in business. ----- Original Message ----- From: To: Sent: Saturday, September 11, 2004 3:03 PM Subject: [Histonet] Being laid off > > My last company went through this twice in the past year. Although they were for profit, some things to consider is if it is an "at will" employment forget the 60 days, cobra is an entitlement based on if you had coverage and is a Federal Law they will have to abide by(plus you can get it yourself), the retirement annuities issues is based on vesting times established at the time of employment, and unused vacation was paid(by law), no sick time or floating days were paid, and severance was not a given for everyone, some folks got nothing. This is wwhat happened at this place, a pharma company. > > > Nick(Rocky) Madary, HT,HTL(ASCP)QIHC > Histology Manager, Medimmune Inc > Cell-3012334950 > Work-3013984745 > Fax-3013989745 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From JColCLEFA <@t> aol.com Sat Sep 11 17:37:43 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] IHC Formalin vs frozen Message-ID: My lab works like most clinical labs in that all tissue is submitted in fixative for H&E staining first. Once the pathologist sees the H&E sections he or she will determine whether special stains or Immunostains are required. We process about 100,000 surgical cases per year, and I do about 400 Immuno slides a day. It is much easier and more efficient to work with paraffin blocks rather than cut 400 frozen slides each day for our 5 hour TAT requirement. I've worked with frozen and paraffin processed material extensively, (I also run our muscle biopsy lab) and as a muscle tech I assume you realize how labor intensive it would be to handle such a high volume of varied tissue (bone marrow biopsies, fatty and fibrous breast tissue, cytology specimens etc) in a frozen arena. We also recieve breast tissue and colon tumor tissue from other hospitals and from as far back at 1980 and the standard of care is formalin fixation and paraffin processing. We have done intensive comparison studies (frozen to fixed) and I can assure all my pathologists that our staining reactions are optimal. In our high volume, low staffing (me and 1 other tech) situation, it's easier to keep as many procedures as similar as possible. Also, lastly, we cut 2 micron sections for 40% of our slides and the distortion from freezing+ the thinness of section required eliminate the utility of frozen sectioning. From BADS27 <@t> msn.com Sat Sep 11 23:21:43 2004 From: BADS27 <@t> msn.com (BETH DELESCAVAGE) Date: Fri Sep 16 15:24:01 2005 Subject: [Histonet] Re: Histonet Digest, Vol 10, Issue 16- New question Message-ID: Hello Everyone- We are starting up an IHC department in a Vet. Lab and was wondering if animal tonsils or lymph nodes work for T and B cell markers, or what is working in your animal lab for CD3, CD45, and CD79a? Thanks in advance. -Beth Delescavage, BS, HTL Phoenix Central Labs Everett, WA ----- Original Message ----- From: histonet-request@lists.utsouthwestern.edu Sent: Saturday, September 11, 2004 10:03 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 10, Issue 16 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: Question regarding 10% Neutral buffered formalin/Immuno (Barry R Rittman) 2. (no subject) (Giorgia Setti) 3. Nemesis stainer (Olszewski, Dawn) 4. Re: Tissue transport from processor to embedding station (Amy Porter) 5. Work load for research labs doing GMA (Macke, Gail) 6. IHC on Frozen Sections (Allen, Rhonda) 7. GFP in a fixed brain tissue (Ajai Vyas) 8. Re: Leica 1800 cryostat manual (Don.Birgerson@leica-microsystems.com) 9. Off Topic - question about class action suit against your "employer" (Victoria Baker) ---------------------------------------------------------------------- Message: 1 Date: Fri, 10 Sep 2004 12:16:58 -0500 From: "Barry R Rittman" Subject: RE: [Histonet] Question regarding 10% Neutral buffered formalin/Immuno To: "histonet" Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0F13257@UTHEVS3.mail.uthouston.edu> Content-Type: text/plain; charset="us-ascii" The original idea of adding formalin or glutaraldehyde to the EDTA was to prevent maceration during the long exposure to EDTA solutions. In my experience these aldehydes significantly decrease the rate of EDTA demineralization. This may well be due to increased cross linking, especially if the original fixation was of relatively short duration. Have no evidence or references for this. My advice would be to use EDTA pH7.2 to 7.4 to eliminate this as a potential problem. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dunn-Jena, Patsy A Sent: Friday, September 10, 2004 9:49 AM To: Histonet Subject: [Histonet] Question regarding 10% Neutral buffered formalin/Immuno I have a graduate student who has come to me with a question I cannot answer as I have limited Immunohistochemical experience. One of her advisers wants her to go to using an EDTA decalcification recipe which contains the 10% NBF in it which is a recipe we used on all of his previous work. She asked me about the NBF crosslinking her immuno sites as she has been using an EDTA decalcification only recipe. Some input here would be nice so I can better explain it to her. Patsy A. Dunn-Jena, RVT, LAT, HT (ASCP) Mineralized Tissue and Histology Research Laboratory Indiana University School of Dentistry 1121 W. Michigan Street, Room 238 Indianapolis, IN 46202 padunnje@iupui.edu (317)274-0544 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Fri, 10 Sep 2004 18:23:54 +0100 From: Giorgia Setti Subject: [Histonet] (no subject) To: histonet@pathology.swmed.edu Message-ID: <1094837034.4141e32a9fa7a@impmail.kcl.ac.uk> Content-Type: text/plain; charset=ISO-8859-1 -- Giorgia Setti giorgia.setti@kcl.ac.uk hello histonetters!!!! i am desperate because I can't find a good antibody for MCP-1 staining on rat frozen sections;I tried the MCP-1 antibody of Abcam but it didn't work;do you know any other antibody tested on frozen section?? thank you Giorgia ---------------------------------------------------------------- This message was sent using IMP, the Internet Messaging Program. ------------------------------ Message: 3 Date: Fri, 10 Sep 2004 13:29:18 -0400 From: "Olszewski, Dawn" Subject: [Histonet] Nemesis stainer To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <825F2CDD64ACF34D85E5E5CCC9EA01320557EA2B@email.sgmc.org> Content-Type: text/plain Does anyone have any positive or negative comments about the Nemesis IHC stainer? Any info would be appreciated. Thanks in advance. Dawn Olszewski HTL(ASCP)QIHC South Georgia Medical Center ------------------------------ Message: 4 Date: Fri, 10 Sep 2004 10:45:54 -0700 (PDT) From: Amy Porter Subject: Re: [Histonet] Tissue transport from processor to embedding station To: Maria Christensen , histonet@lists.utsouthwestern.edu Message-ID: <20040910174554.28637.qmail@web40901.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Maria - reply to me (off list) if you want and supply me with your fax number and i can fax you pictures of what we have here. It is a Thermal Console we have model #4585 from Miles/Sakura. We also walk several feet to our embedding station and always have just placed our cassettes into the reservoir then let them sit for a few minutes to rewarm. They usually do not get that cold in that short of a time span. Maria Christensen wrote: Thanks to all who replied. To answer some of your questions, our processor and embedding center are in separate rooms now, but not far from each other (10-12 steps). Our concern was keeping the tissue warm so that we could embed quickly when needed without having to warm the tissue up. We thought of just transferring in a beaker of melted paraffin, but sometimes we can't get to all the cassettes right away, so the wax would begin to solidify. Unfortunately we don't have an oven in the lab where the embedding equipment is. Our Tissue Tek II embedding station has a heated compartment for the molds, but no compartment for the cassettes. The manual mentions an infiltration process carrier, and a vacuum infiltrator, but these accessories did not come with our reconditioned equipment. I would like to know more about the holding reservoir that Amy Porter mentioned - part no., where purchased, etc. because if the basket from the VIP fits in it, that would be ideal. Again, thanks for your ideas- I'm a firm believer in not reinventing the wheel! Maria Maria A. Christensen Technical Associate Medical Microbiology & Immunology Creighton University School of Medicine 2500 California Plaza Omaha, NE 68178-0404 phone (402) 280-2678 FAX (402) 280-1875 email mariac@creighton.edu _______________________________________________ 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!? Yahoo! Mail Address AutoComplete - You start. We finish. ------------------------------ Message: 5 Date: Fri, 10 Sep 2004 14:06:03 -0400 From: "Macke, Gail" Subject: [Histonet] Work load for research labs doing GMA To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Dear Histonetters, Could some one tell me what is considered the average work load for cutting GMA blocks. How many Blocks per year embedded? How many slides cut per year and stained per year? We use Technovit 7100 for processing and embedding. We are a Core Research Lab that also does paraffin and frozen processing and cutting. Most of the information I have found relates to Clinical Labs doing paraffin and frozen work. I haven't found much related to Research Labs. Any information would be helpful. Thank you for taking the time to read and reply to my questions. Gail Macke, HTL Shriners Hospital for Children: Shriners Burns Hospital- Cincinnati Cincinnati, Ohio, USA Gmacke@Shrinenet.org CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. ------------------------------ Message: 6 Date: Fri, 10 Sep 2004 12:30:45 -0700 From: "Allen, Rhonda" Subject: [Histonet] IHC on Frozen Sections To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <707BC118BC44D3118E3B0000F81F1E7605A393DA@VHAKANEXC1> Content-Type: text/plain Hello, I am trying to do the T & B cells on frozen section tonsil for my IHC exam due at the end of this month. I have tried multiple procedures, and have not yet found one that makes me happy, or looks like the one in the picture at the IHC home page. If someone could e-mail me their procedures (mostly from cutting to antibody application) it would be greatly appreciated. By the way, my cryostat will not cut thinner than 4-6 microns. Thanks in advance, Rhonda Allen VA Medical Center ------------------------------ Message: 7 Date: Fri, 10 Sep 2004 18:12:43 -0700 From: Ajai Vyas Subject: [Histonet] GFP in a fixed brain tissue To: histonet@lists.utsouthwestern.edu Message-ID: <934be6370409101812725ddc95@mail.gmail.com> Content-Type: text/plain; charset=US-ASCII Hi, Regards from a new entrants to histonet. I have GFP labelled parasites sitting in brain of my rats. I need to slice the brain and locate these parasites. Will GFP still be present if I use fixatives like formaldehyde or organic solvents like xylene? What fixing protocols are best for preserving GFP signal? Thanks in advance, AJAI ------------------------------ Message: 8 Date: Fri, 10 Sep 2004 11:20:57 -0500 From: Don.Birgerson@leica-microsystems.com Subject: Re: [Histonet] Leica 1800 cryostat manual To: "Amy Self" Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Hi Amy, I be happy to mail you a copy of the "User Manual" if you will give me your street address. Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Amy Self" Sent by: To: histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] Leica 1800 cryostat manual 09/10/2004 09:25 AM Does anyone have a manual for the old Leica 1800 cryostat that they could share with me or don't need anymore? We (biomed dept.) has misplaced ours. Thanks, amy NOTE: The information contained in this message may be privileged, 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 this message and deleting it from your computer. Thank you. _______________________________________________ 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 ______________________________________________________________________ ------------------------------ Message: 9 Date: Sat, 11 Sep 2004 06:53:05 -0700 (PDT) From: Victoria Baker Subject: [Histonet] Off Topic - question about class action suit against your "employer" To: HistoNet Server Message-ID: <20040911135305.17671.qmail@web50105.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Good morning, This is a question that I hope very few of us on the list-server have ever had to, are currently, or will ever have to deal with/answer personally. A week ago yesterday the not-for-profit/NCI (Cancer Center Support Grant)funded organization I work for gave us WARN notification that told us we would be permanently "layed-off" as of Sept. 10 unless an agreement between our facility and the government was made. As negotiations were being worked for a merger with "other" institutions there might still be a way to work out our difficulties and keep our institution open. The specifics of this merger are unknown and we have had no specific information given to us. As of yesterday afternoon we were informed that we would have to call in on Monday morning to see if we would be allowed access to the building and work. The controversy is this: 1. According to law we are to be notified at least 60 days in advance of a permanent closing. This has not been done. 2. We lose any severance or unused vacation pay still owed to us under a "normal" - I use this term loosely - closing. 3. Insurance benefits - being able to apply for COBRA or get clear information as to what we could do to purchase insurance on our own has been non existent and our HR person left her position as of Friday as well. This is not part of the actual legal proceedings just concerns of all employees. 4. Retirement plans/annuities are a separate issue as not all employees were involved in these benefits. We are a very small institute of less than 150 people in NYS. I'm looking to see if anyone else on the server has had this occur to them and whether or not they did pursue a legal action to get the benefits due to them. I'd also like to know if anyone has information or people/places I could contact about what most have gotten in terms of litigation. The issues I listed are the most pertinent, as with many of these closings other issues revolve around the main ones. Many of these people have been here since the late 70's or early 80's. Yesterday as most of us were cleaning out offices and trying to organize our labs, I encountered so many different human reactions and emotions. When I left my lab last night it was just around sundown and shadows were cast over some of my equipment and benches after I had shut down the lights (I have a wall of windows that I've always loved to watch the seasons or the weather)and it was a very sad moment. Would I ever see it again? When you are layed off or a company closes it's a 'done deal' and you can move on. When you just seem to get a 'stay of termination' that leaves you no where closer to resolution it wears you down. If anyone has anything that they could pass on I would very much appreciate it. Thank you very much. Vikki Baker Institute for Cancer Prevention Valhalla, NY 10595 __________________________________________________ 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 End of Histonet Digest, Vol 10, Issue 16 **************************************** From c.m.vanderloos <@t> amc.uva.nl Sun Sep 12 08:32:28 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Re: chromogen Message-ID: <1a5cd61a3ca1.1a3ca11a5cd6@amc.uva.nl> Dear Patsy, Gayle is completely right about the Permanent Red as Alk phos chromogen. I regard it as crisp as DAB, perhaps even better. If you wish to stick to HRP, indeed Vector Nova Red is a very good choice too. I would add that this substrate needs to be mounted fully organic, being an advantage over AEC. There is also an HRP chromogen called Romulin Red from BioCare. The colored reaction product appears to be identical to Vector Nova Red and perhaps they are chemically identical. Hope this addition is helpful, Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Gayle Callis Date Wed, 08 Sep 2004 13:29:26 -0600 To "Patsy Ruegg" , Histonet@lists.utsouthwestern.edu Subject Re: [Histonet] chromogen DAKO has Permanent Red for alk phos IHC. Chris van der Loos indicated DAKO product was very sensitive, and it also fluoresces. Vector red is AP and also fluoresces. Have not tried Vector Red to compare with primary antibody concentrations. These reds are more on dark pink red side, rather than orangish red of AEC, or Brick red of NOVA Red. NOVA Red from Vector, but it is a brick red!! Darker than AEC. You can mount AEC from water, using Crystal mount, let this dry and then add a permanent mounting media. At 11:39 AM 9/8/2004, you wrote: >What is a good substitute for DAB using HRP, my investigator wants >prettier red or green colors for a poster, something like he gets with >FITC, but I don't want to do flourescence? I know I could use AEC but I >would prefer something more permanent. Didn't Kirkgaurd and Perry come >out with a permanent red chromogen for HRP? >Patsy >_______________________________________________ >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 From c.m.vanderloos <@t> amc.uva.nl Sun Sep 12 08:37:51 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] RE: PCNA - URINARY BLADDER Message-ID: <11af7211ed40.11ed4011af72@amc.uva.nl> Dear Dr. Balaji, The type of problem as you describe for anti-PCNA sounds familiar to me. We changed to the new rabbit anti-Ki67 monoclonal antibody from NeoMarkers/LabVision. That antibody works perfectly for human, mouse and rat tissues. Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From balajimr@drreddys.com Date Thu, 09 Sep 2004 12:04:53 +0530 To histonet@lists.utsouthwestern.edu Subject [Histonet] PCNA - URINARY BLADDER Dear Histonetters, It's nice to be back with Histosearch after a short break. Well, I am facing problem with PCN staining of urinary bladder of rat. We are using Zymed PCNA staining kit for the same. I am getting non specific staining where even the nucleus of the muscle layers are also staining. We are retrieving the antigen by heating the sections in Citrate buffer. But we are not having this problem in the intestines of same control animals. Does anybody have experience of PCNA staining with U. bladder so that they can advise me about this problem. Ofcourse we have increased the blocking time with serum upto 30 minutes unsuccessfully. Thanks in advance. Dr. M.R. Balaji Dept. Pre clinical safety evaluation, Discovery research, Dr. Reddys Laboratories ltd. Bollaram Road, Miyapur, Hyderabad, 500 049 Andhra Pradseh, INDIA Email - balajimr@drreddys.com Tel: 040- 23045439 - Ext.432. From jlinda <@t> ces.clemson.edu Sun Sep 12 12:29:23 2004 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Super tech??? Message-ID: <5.2.1.1.2.20040912132002.028a0490@mailhost.ces.clemson.edu> Dear , To quote your recent post: "We process about 100,000 surgical cases per year, and I do about 400 Immuno slides a day." PLUS you do muscle biopsies AND with only 2 techs. Now, granted, it has been 12 years since I've been in clinical practice...but...these numbers seem phenomenal to me! Please tell me that your lab is completely automated from IHC stainer to coverslipper to bar coding???? If not, please send name of vitamins you are currently taking! Thanks, Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo/histo.htm From miller <@t> coho.net Sun Sep 12 12:32:23 2004 From: miller <@t> coho.net (Diane G. Miller) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Karen Meirs Message-ID: <044f01c498ee$799df550$0300a8c0@desktop> Hello Everyone, I'm looking for Karen Meirs, if you are out there, would you please contact me. If anyone knows where I can contact Karen would you please forward me her information. Thanks Diane 503-784-6444 miller@coho.net From dlcowie <@t> prodigy.net Sun Sep 12 14:07:44 2004 From: dlcowie <@t> prodigy.net (Dawn Cowie) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] (no subject) Message-ID: <20040912190744.42190.qmail@web81003.mail.yahoo.com> hi netters, I am looking for Gerri Goforth, a travelling tech. If you're still out there, please respond. You worked for Ameripath for about a year in Clearwater a couple of years ago. I have tried to contact you through the temp agencies but with no luck. If anyone knows where she is please let me know. From jnocito <@t> satx.rr.com Sun Sep 12 19:09:32 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Off Topic - question about class action suit against your "employer" References: <20040911135305.17671.qmail@web50105.mail.yahoo.com> Message-ID: <018e01c49925$f22817d0$4661ce44@yourxhtr8hvc4p> Victoria a couple of years ago, a private lab locked their doors over the weekend and did not let anyone know. The employees showed up to work on Monday just to find that the doors were chained and they had no jobs. Now, I realize this is a private lab, but there was not a single thing they could do. Several companies had thousands of dollars outstanding with this company. The government auctioned off everything that was in the building and took what was theirs first. Of course, there was nothing left over for the several supply companies, utilities, building management or reference labs like mine. You can sue, but what will Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX you get if they have no assists? ----- Original Message ----- From: "Victoria Baker" To: "HistoNet Server" Sent: Saturday, September 11, 2004 8:53 AM Subject: [Histonet] Off Topic - question about class action suit against your "employer" > Good morning, > > This is a question that I hope very few of us on the > list-server have ever had to, are currently, or will > ever have to deal with/answer personally. > > A week ago yesterday the not-for-profit/NCI (Cancer > Center Support Grant)funded organization I work for > gave us WARN notification that told us we would be > permanently "layed-off" as of Sept. 10 unless an > agreement between our facility and the government was > made. As negotiations were being worked for a merger > with "other" institutions there might still be a way > to work out our difficulties and keep our institution > open. The specifics of this merger are unknown and we > have had no specific information given to us. As of > yesterday afternoon we were informed that we would > have to call in on Monday morning to see if we would > be allowed access to the building and work. > > The controversy is this: > 1. According to law we are to be notified at least 60 > days in advance of a permanent closing. This has not > been done. > 2. We lose any severance or unused vacation pay still > owed to us under a "normal" - I use this term loosely > - closing. > 3. Insurance benefits - being able to apply for COBRA > or get clear information as to what we could do to > purchase insurance on our own has been non existent > and our HR person left her position as of Friday as > well. This is not part of the actual legal > proceedings just concerns of all employees. > 4. Retirement plans/annuities are a separate issue as > not all employees were involved in these benefits. > > We are a very small institute of less than 150 people > in NYS. > > I'm looking to see if anyone else on the server has > had this occur to them and whether or not they did > pursue a legal action to get the benefits due to them. > I'd also like to know if anyone has information or > people/places I could contact about what most have > gotten in terms of litigation. The issues I listed are > the most pertinent, as with many of these closings > other issues revolve around the main ones. > > Many of these people have been here since the late > 70's or early 80's. Yesterday as most of us were > cleaning out offices and trying to organize our labs, > I encountered so many different human reactions and > emotions. When I left my lab last night it was just > around sundown and shadows were cast over some of my > equipment and benches after I had shut down the lights > (I have a wall of windows that I've always loved to > watch the seasons or the weather)and it was a very sad > moment. Would I ever see it again? When you are layed > off or a company closes it's a 'done deal' and you can > move on. When you just seem to get a 'stay of > termination' that leaves you no where closer to > resolution it wears you down. > > If anyone has anything that they could pass on I would > very much appreciate it. > > Thank you very much. > > Vikki Baker > Institute for Cancer Prevention > Valhalla, NY 10595 > > > > > > __________________________________________________ > 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 JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk Mon Sep 13 04:05:19 2004 From: JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk (JOHN PHILLIPS) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] IHC Formalin vs frozen Message-ID: <166A1E642B5B644DA694C08FD29D0ADC3E1265@ztroy.new-tr.wales.nhs.uk> Hello fellow "Histotech". Do you find the time with this workload to do the QC as well?!!! Your workload shared between two is phenomenal.How many arms have you got? If you have managed to master the application of translocation could you please share this with the rest of us in the real world. You must go through tons of anti-perspirant or is this a slight exageration on my part? Oh and please do not share your work practice secrets with NHS management here in the UK. Super techs like you would put a lot of lab staff out of work. John. > ---------- > From: JColCLEFA@aol.com[SMTP:JColCLEFA@aol.com] > Sent: 11 September 2004 23:37 > To: histonet-request@lists.utsouthwestern.edu; histonet@lists.utsouthwestern.edu; georgecole@ev1.net > Subject: [Histonet] IHC Formalin vs frozen > > My lab works like most clinical labs in that all tissue is submitted in > fixative for H&E staining first. Once the pathologist sees the H&E sections he or > she will determine whether special stains or Immunostains are required. We > process about 100,000 surgical cases per year, and I do about 400 Immuno slides a > day. It is much easier and more efficient to work with paraffin blocks rather > than cut 400 frozen slides each day for our 5 hour TAT requirement. I've > worked with frozen and paraffin processed material extensively, (I also run our > muscle biopsy lab) and as a muscle tech I assume you realize how labor > intensive it would be to handle such a high volume of varied tissue (bone marrow > biopsies, fatty and fibrous breast tissue, cytology specimens etc) in a frozen > arena. We also recieve breast tissue and colon tumor tissue from other hospitals > and from as far back at 1980 and the standard of care is formalin fixation and > paraffin processing. We have done intensive comparison studies (frozen to > fixed) and I can assure all my pathologists that our staining reactions are > optimal. In our high volume, low staffing (me and 1 other tech) situation, it's > easier to keep as many procedures as similar as possible. Also, lastly, we cut 2 > micron sections for 40% of our slides and the distortion from freezing+ the > thinness of section required eliminate the utility of frozen sectioning. > _______________________________________________ > 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 gcallis <@t> montana.edu Mon Sep 13 09:54:03 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] GFP in a fixed brain tissue In-Reply-To: <934be6370409101812725ddc95@mail.gmail.com> References: <934be6370409101812725ddc95@mail.gmail.com> Message-ID: <6.0.0.22.1.20040913085128.01b28c60@gemini.msu.montana.edu> Maybe, it is probably best to cut frozen sections, fix with 2% paraformaldehyde for 1 minute, rinse, and mount with Molecular Probes, Probe On antifade mounting media, ready to use. Go to Clontech website and read Living Colours manual on some of these issues. Their tech services is excellent. This question has been discussed, even recently, on Histonet, you can search the archives at www.histosearch.org At 07:12 PM 9/10/2004, you wrote: >Hi, > Regards from a new entrants to histonet. I have GFP labelled >parasites sitting in brain of my rats. I need to slice the brain and >locate these parasites. Will GFP still be present if I use fixatives >like formaldehyde or organic solvents like xylene? What fixing >protocols are best for preserving GFP signal? >Thanks in advance, >AJAI > >_______________________________________________ >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 bwhitaker <@t> brownpathology.com Mon Sep 13 10:23:26 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] FISH fading Message-ID: <000001c499a5$9e18b750$3601a8c0@brownpathology.net> Hi Histonet! A colleague called to vent some frustrations about FISH today. This person is performing FISH, checking the slides, and thinking the results are good. A few days later when the PI checks the slides, the slides have faded. Is there a specific rule of thumb for how much exposure to light (sunlight and artificial) and how much time can pass without damaging the signal? Are anti-fading agents different? Should he try something other than what the manufacturer includes in the kit that they are using? Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 From galinadeyneko <@t> yahoo.com Mon Sep 13 10:51:14 2004 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] embedding of small artery Message-ID: <20040913155114.4493.qmail@web14523.mail.yahoo.com> Dear Colleagues' Could someone share an experience about embedding of tissue in agarose gel with further embedding in OCT and cryostate sectioning and , maybe, paraffin processing. ( I found very small piece of information about agarose gel in Myneurolab.com). Our goal is to embed fresh murine common carotid artery and receive ring (circle) of cross sectioned artery on slide. I per fuse intracardial PBS, 10 % formalin, and PBS+OCT in ratio 1:1, after freeze on dry ice harvested artery, but this very small artery still very soft ant thaws momentarily for embedding straight. Any advices will appreciate. Galina Deyneko Novartis, Cardiovascular department Cambridge,MA ph: 617-871-7613 --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - Send 10MB messages! From DDittus787 <@t> aol.com Mon Sep 13 12:48:40 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Re: Anti-parrafin floor Message-ID: <7B0A3F7A.56A40CDC.0A1F969F@aol.com> Barb: the flooring came from Acme Panels-the company is in the UK and can be found on Google, it is anti-solvent, chemical, paraffin,etc. It comes in 3 or 4 colors and everyone loves it. We had a contractor bring it in and install it. dana From carl.hobbs <@t> kcl.ac.uk Mon Sep 13 13:02:14 2004 From: carl.hobbs <@t> kcl.ac.uk (carl hobbs) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] re embedding of small artery Message-ID: In my experienc, with fixed tissues it is easy to embed using agarose, then to re-orient the tissue for frozen/pwax sectioning: Dissolve 1% agarose( aq) and when about 50C place the tissue in the agarose, in a bigger mould. It will set quickly.....better if placed at 4C for a while. Remove the set agarose and cut away excess agarose so you have the tissue in the orientation of your choice. Then process to pwax. Or, place in OCT for a couple of hours on a rotator. Then embed in an OCT filled mould in the orientation you need and....freeze. If you are doing fresh tissue, get the melted, cooling agarose into a mould , place the tissue and cool quickly. Immediately place in OCT, swirl gently for a minute, then freeze. Play around with control material until you are confident of the technique and to reassure yourself that the elevated temp does not adversely afffect your tissue. You can also do gelatin/vibratome sections of the fixed tissue, if you have a vibratome. --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.754 / Virus Database: 504 - Release Date: 06/09/2004 From k_byrnes <@t> yahoo.com Mon Sep 13 13:34:29 2004 From: k_byrnes <@t> yahoo.com (Kim Byrnes) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Recover tissue sections on non-plus slides Message-ID: <20040913183429.84652.qmail@web12308.mail.yahoo.com> Does anyone have a method or know of a product that can recover or rescue tissue sections that were accidentally mounted onto non-plus (non-polarized, non-coated) slides? We would like to do immuno on these slides, but they consistently fall off of the slides. Any ideas? Thanks, Kim Georgetown University __________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail From bwhitaker <@t> brownpathology.com Mon Sep 13 14:24:14 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Recover tissue sections on non-plus slides In-Reply-To: <20040913183429.84652.qmail@web12308.mail.yahoo.com> Message-ID: <000301c499c7$41758920$3601a8c0@brownpathology.net> There is a procedure to remove tissue sections from a slide, and they can then be mounted on + slides. The product that works best in my hands is Mount-Quick. It has more than one distributer, but I know that Newcomer Supply (800-383-7799) has this product, as well as directions on how to do the procedure. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Byrnes Sent: Monday, September 13, 2004 1:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Recover tissue sections on non-plus slides Does anyone have a method or know of a product that can recover or rescue tissue sections that were accidentally mounted onto non-plus (non-polarized, non-coated) slides? We would like to do immuno on these slides, but they consistently fall off of the slides. Any ideas? Thanks, Kim Georgetown University __________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From chiggerson <@t> memhosp.com Mon Sep 13 14:46:54 2004 From: chiggerson <@t> memhosp.com (chiggerson@memhosp.com) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Recover tissue sections on non-plus slides In-Reply-To: <20040913183429.84652.qmail@web12308.mail.yahoo.com> Message-ID: Kim, The 2000 Tech Sample exercise HT-1 gives a procedure for this. The title of the exercise is "Use Mount Quick to Remove Tissue Sections from Non-Positively Charged Slides for Immunohistochemical and Special Stains". Good luck! Cindy Cindy Higgerson HTL(ASCP) Surgical Pathology Supervisor Memorial Hospital Kim Byrnes Sent by: histonet-bounces@lists.utsouthwestern.edu 09/13/2004 01:34 PM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] Recover tissue sections on non-plus slides Does anyone have a method or know of a product that can recover or rescue tissue sections that were accidentally mounted onto non-plus (non-polarized, non-coated) slides? We would like to do immuno on these slides, but they consistently fall off of the slides. Any ideas? Thanks, Kim Georgetown University __________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From julien_lambreydesouza <@t> uqar.qc.ca Mon Sep 13 15:58:59 2004 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] alizarin red fading Message-ID: <00c501c499d4$7dc51140$9310d784@uqar.qc.ca> Hello all, We do differencial staining of bone and cartillage on small fish (~1cm). To do so, we use the Clear and Stain technique which results in red bone coloration and blue cartillage coloration. After a month or so, We noticed that the red stain tends to fade off. We tried to re-stain the fish, but the alizarin red wouldn't stain the bones anymore. Why is this so? Is it that bone decalcifies in the glycerin? We put a few cristals of thymol in the glycerin to avoid mold development. Could this be a decalcifying agent? Does anybody know of a way to re-stain? For the time being, we score bony elements within 3 days of staining to avoid fading problems, but this stops us from doing batch staining. Batch staining would be much faster. We would then keep stained fish in glycerin until scoring. Thanks for any suggestion. Julien Lambrey de Souza Biologie ?volutive, Universit? du Qu?bec ? Rimouski, Julien Lambrey de Souza Biologie ?volutive, Universit? du Qu?bec ? Rimouski, (418) 723-1986 #1438 From Xudong_Cao <@t> brown.edu Mon Sep 13 16:03:50 2004 From: Xudong_Cao <@t> brown.edu (Cao, Xudong) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] fluorescent dyes/lables for neuronal activities? Message-ID: <1DA88DDC48CCC245A777599FDAEED6D0A3909B@MAIL1.AD.Brown.Edu> Greetings! first of all I'd like to thank eveybody who responded to my previous posts. The advice/ideas I received were valuable! Here I have one more question: I am studying nerve innervation into artificial skin graft that we make in vitro and my immunostaining tells me that there are some wonderful nerve fibers growing into the skin. The question that we have now is if the innervation functional. In order to test this, we want to mechanically tease the artificial skin and see if there is some responses from the nerve cells that have presumablly innervated the corresponding part of the skin. Thus, it would be nice to find some fluorescent dyes/lables that will glow when the neurons are activated/firing. any suggestions? best regards, Xudong From Barry.R.Rittman <@t> uth.tmc.edu Mon Sep 13 16:29:32 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] alizarin red fading Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0F13357@UTHEVS3.mail.uthouston.edu> I cannot see why the addition of a few crystals of thymol would make the glycerin acidic enough to cause removal of calcium and thus leaching of the alizarin. If you are using 100% glycerin, there should be no need for the addition of thymol. I am not sure exactly how you have processed these specimens. If you are using the classical method with potassium hydroxide (as the leaching agent to remove excess stain from soft tissues), is it possible that there were traces of potassium hydroxide left in the specimen? I have some specimens that were stained in 1961 and some in 1970 that show no signs of fading. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Julien Sent: Monday, September 13, 2004 3:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] alizarin red fading Hello all, We do differencial staining of bone and cartillage on small fish (~1cm). To do so, we use the Clear and Stain technique which results in red bone coloration and blue cartillage coloration. After a month or so, We noticed that the red stain tends to fade off. We tried to re-stain the fish, but the alizarin red wouldn't stain the bones anymore. Why is this so? Is it that bone decalcifies in the glycerin? We put a few cristals of thymol in the glycerin to avoid mold development. Could this be a decalcifying agent? Does anybody know of a way to re-stain? For the time being, we score bony elements within 3 days of staining to avoid fading problems, but this stops us from doing batch staining. Batch staining would be much faster. We would then keep stained fish in glycerin until scoring. Thanks for any suggestion. Julien Lambrey de Souza Biologie ?volutive, Universit? du Qu?bec ? Rimouski, Julien Lambrey de Souza Biologie ?volutive, Universit? du Qu?bec ? Rimouski, (418) 723-1986 #1438 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Mon Sep 13 16:43:49 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Recover tissue sections on non-plus slides Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5C062@EXCHANGE1.huntingtonhospital.com> We have used the Mount Quick from Newcomer Supply, and it works well. Laurie Colbert Huntington Memorial Hospital Pasadena, CA -----Original Message----- From: Kim Byrnes [mailto:k_byrnes@yahoo.com] Sent: Monday, September 13, 2004 11:34 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Recover tissue sections on non-plus slides Does anyone have a method or know of a product that can recover or rescue tissue sections that were accidentally mounted onto non-plus (non-polarized, non-coated) slides? We would like to do immuno on these slides, but they consistently fall off of the slides. Any ideas? Thanks, Kim Georgetown University __________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From robint <@t> mediom.qc.ca Mon Sep 13 19:01:58 2004 From: robint <@t> mediom.qc.ca (Robin Turcotte) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] cryomacrocut Message-ID: <000701c499ee$cb688f70$df8ceccf@client> Does anyone have an address from a compagny or an hospital who have a used cryomacrocut in good condition to be sold. Thank you Robin From robint <@t> mediom.qc.ca Mon Sep 13 19:07:19 2004 From: robint <@t> mediom.qc.ca (Robin Turcotte) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] cryomacrocut Message-ID: <000e01c499ee$ee9cdbe0$df8ceccf@client> Does anyone have an address from a compagny or an hospital who have a used cryomacrocut in good condition to be sold. Thank you Robin From robint <@t> mediom.qc.ca Mon Sep 13 19:07:47 2004 From: robint <@t> mediom.qc.ca (Robin Turcotte) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] cryomacrocut Message-ID: <000f01c499ee$ef064c10$df8ceccf@client> Does anyone have an address from a compagny or an hospital who have a used cryomacrocut in good condition to be sold. Thank you Robin From robint <@t> mediom.qc.ca Mon Sep 13 19:10:32 2004 From: robint <@t> mediom.qc.ca (Robin Turcotte) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] cryomacrocut Message-ID: <001301c499ef$45100dd0$df8ceccf@client> Does anyone have an address from a compagny or an hospital who have a used cryomacrocut in good condition to be sold. Thank you Robin From robint <@t> mediom.qc.ca Mon Sep 13 19:11:36 2004 From: robint <@t> mediom.qc.ca (Robin Turcotte) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] cryomacrocut Message-ID: <001c01c499ef$6fce96e0$df8ceccf@client> Does anyone have an address from a compagny or an hospital who have a used cryomacrocut in good condition to be sold. Thank you Robin From jstaruk <@t> masshistology.com Mon Sep 13 20:15:52 2004 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Shandon Hypercenter Xp In-Reply-To: Message-ID: <000201c499f8$87d3cb80$6401a8c0@yourw04gtxld67> I'm looking for someone familiar with the Shandon Hypercenter Xp tissue processor. I need some technical advise. Can someone help me? Anyone planning to invoice me for their help need not reply. Thank you Jim ________________________ James E. Staruk, HT(ASCP) Mass Histopathology Service www.masshistology.com From bills <@t> icpmr.wsahs.nsw.gov.au Mon Sep 13 22:43:17 2004 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Superfrost Slides Message-ID: <000101c49a0c$f9356e50$e1ce080a@wsahs.nsw.gov.au> Is anyone having troubles with NO staining on some slides on the Ventana Benchmark. The biggest problem seems to be with the EDTA buffered CC1 retrieval solution. Not all slides are effected, just an occasional one, but we usually end up with little or no staining with anything including the Haematoxylin. We use Menzel Superforst Plus and Superfrost Plus Ultra, and both slides have shown the effect. Thanks Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. 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 raghul <@t> sctimst.ker.nic.in Tue Sep 14 06:01:15 2004 From: raghul <@t> sctimst.ker.nic.in (Dr. Raghul) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] embedding of small artery_paraffin Message-ID: hello galina, Rat and mouse organs can be cut easily after short processing(using acetone and xylene)and then embedding in paraffin wax of low melting point (56 C). We have followed the protocols from below given reference successfully and same could applied for the small artery also Theory and practice of Histological techniques (bancroft and steven) (or) Manual of histologic staining methods of the armed forces institutes of pathology edited by Lee G. Luna, III edition page 16 Raghul J Histopathology-Implant Biology Sree chitra tirunal institute Thiruvananthapuram 695 012 From raghul <@t> sctimst.ker.nic.in Tue Sep 14 06:26:23 2004 From: raghul <@t> sctimst.ker.nic.in (Dr. Raghul) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] vascular graft_sectioning difficulty Message-ID: hello members, We cut 5 ?m thin sections of paraffin embedded vascular graft(Dacron) in a rotary microtome with disposable blades but the senior tech tells that the tissue sections are crumbling and forces us to go for thick sections around 10?m but again not with much success. We can understand that there is not enough tissue adhereing to the graft and we are literally cutting the graft. But did anyone had such problems sectioning vascular grafts. Any suggestions? regards , Raghul J scientist Histopathology- implant biology division Srichitra tirunal institute of medical sciences and technology Trivandrum -695 012 Kerala, India From nena.dimaano <@t> stryker.com Tue Sep 14 06:54:55 2004 From: nena.dimaano <@t> stryker.com (Dimaano, Nena) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Test Message-ID: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD06210EE4@HOS2KEXCHCL.howost.strykercorp.com> Nena Dimaano Advanced Technology Stryker Orthopaedics 325 Corporate Drive Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com From JNocito <@t> Pathreflab.com Tue Sep 14 07:09:09 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Superfrost Slides In-Reply-To: <000101c49a0c$f9356e50$e1ce080a@wsahs.nsw.gov.au> Message-ID: Bill, we've had problems also. It could be that the slides might be too changed, which is causing the reagents to bead up. Currently, we are using positive charged slides from Statlab Medical Products, which I think they get from Erie Scientific. Statlab's number is 1-800-442-3573. The may be able to direct you to some one closer to you. 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 Bill Sinai Sent: Monday, September 13, 2004 10:43 PM To: histonet (E-mail) Subject: [Histonet] Superfrost Slides Is anyone having troubles with NO staining on some slides on the Ventana Benchmark. The biggest problem seems to be with the EDTA buffered CC1 retrieval solution. Not all slides are effected, just an occasional one, but we usually end up with little or no staining with anything including the Haematoxylin. We use Menzel Superforst Plus and Superfrost Plus Ultra, and both slides have shown the effect. Thanks Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From wasielewski.reinhard.von <@t> mh-hannover.de Tue Sep 14 07:29:10 2004 From: wasielewski.reinhard.von <@t> mh-hannover.de (wasielewski.reinhard.von@mh-hannover.de) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Perferred fixative for animals Message-ID: <41470036.30954.8F79EDC3@localhost> Hi Histonetters, We are looking for the most convenient and morpholgical satisfying fixative for mouse and rat tissue beside formaldehyde. We want to stain a wide range of different immunomarkers but don' t like formalin (therefore) too much. Frozen sections are too bad in morphology. Please send me any suggestions ! Many thanks in advance, Reinhard. PD Dr. med. Reinhard von Wasielewski From Kay.Cullen <@t> umassmed.edu Tue Sep 14 07:56:22 2004 From: Kay.Cullen <@t> umassmed.edu (Cullen, Kay) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] RE: Fixative for animals Message-ID: Are you busing straight formaldehyde or formaldehyde/glutaraldehyde? I get very good results with Bouin's for most tissue. Kay Cullen Research Associate Department of Cell Biology University of Massachusetts Medical School Worcester, MA USA From ASelf <@t> gmhsc.com Tue Sep 14 09:05:39 2004 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Histo. Assistant Job Descriptions Message-ID: <39836CD6DB61654E8F95A35898C921860A87F3@exchange.gmhpost.com> Good Morning Netters, We are looking to hire a histology assistant in our lab. I was wandering if anyone had any job descriptions as well as job requirements that they could share with me? We have one but it very minimal as to what the histo assistant should and should not do so our lab manager would like to see what other hospitals are doing and requiring for this job position. Thanks in advance, amy NOTE: The information contained in this message may be privileged, 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 this message and deleting it from your computer. Thank you. From ljb <@t> medicine.wisc.edu Tue Sep 14 09:16:54 2004 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Perferred fixative for animals Message-ID: Dear Reinhard, We use Prefer fixative from Anatech Ltd, www.anatechltdusa. We have beautiful morphology, staining is wonderful (specials and immunos). The only down side of Prefer is that it affects the staining of eosinophils. We've recently learned how to get around that issue in human tissues, but haven't figured it out in rat tissues. Best of luck! Cindy B LaCinda Burchell, BA, AS, HT(ASCP) University of Wisconsin-Madison, Medical School Asthma and Allergy Research IHC Lab 600 Highland Ave. CSC K4/913 Madison, Wisconsin 53792 Phone: 608-262-3518 FAX: 608-263-3746 >>> 09/14/04 07:29AM >>> Hi Histonetters, We are looking for the most convenient and morpholgical satisfying fixative for mouse and rat tissue beside formaldehyde. We want to stain a wide range of different immunomarkers but don' t like formalin (therefore) too much. Frozen sections are too bad in morphology. Please send me any suggestions ! Many thanks in advance, Reinhard. PD Dr. med. Reinhard von Wasielewski _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barbara_Lentz <@t> dahlchase.com Tue Sep 14 09:48:20 2004 From: Barbara_Lentz <@t> dahlchase.com (Barbara Lentz) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Recover tissue sections on non-plus slides Message-ID: We, too, use the Mount Quick from Newcomer Supply. On a regular basis, we remove H&E tissue sections to perform immunos. www.newcomersupply.com is their address, Barb >>> Kim Byrnes 09/13/04 02:34PM >>> Does anyone have a method or know of a product that can recover or rescue tissue sections that were accidentally mounted onto non-plus (non-polarized, non-coated) slides? We would like to do immuno on these slides, but they consistently fall off of the slides. Any ideas? Thanks, Kim Georgetown University __________________________________ Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! http://promotions.yahoo.com/new_mail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kbradshaw <@t> lcpath.com Tue Sep 14 09:41:17 2004 From: kbradshaw <@t> lcpath.com (Kari Bradshaw) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Histo. Assistant Job Descriptions In-Reply-To: <39836CD6DB61654E8F95A35898C921860A87F3@exchange.gmhpost.com> Message-ID: Here is our job description. :) Kari Bradshaw, HT(ASCP) Laboratory Manager Lower Columbia Pathologists 1217 14th Ave Longview, WA 98632 (360)425-5620 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amy Self Sent: Tuesday, September 14, 2004 7:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histo. Assistant Job Descriptions Good Morning Netters, We are looking to hire a histology assistant in our lab. I was wandering if anyone had any job descriptions as well as job requirements that they could share with me? We have one but it very minimal as to what the histo assistant should and should not do so our lab manager would like to see what other hospitals are doing and requiring for this job position. Thanks in advance, amy NOTE: The information contained in this message may be privileged, 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 this message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bwhitaker <@t> brownpathology.com Tue Sep 14 10:54:03 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:24:02 2005 Subject: FW: [Histonet] Histo. Assistant Job Descriptions Message-ID: <000801c49a73$0ee10140$3601a8c0@brownpathology.net> Here is ours. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kari Bradshaw Sent: Tuesday, September 14, 2004 9:41 AM To: Amy Self; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histo. Assistant Job Descriptions Here is our job description. :) Kari Bradshaw, HT(ASCP) Laboratory Manager Lower Columbia Pathologists 1217 14th Ave Longview, WA 98632 (360)425-5620 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amy Self Sent: Tuesday, September 14, 2004 7:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histo. Assistant Job Descriptions Good Morning Netters, We are looking to hire a histology assistant in our lab. I was wandering if anyone had any job descriptions as well as job requirements that they could share with me? We have one but it very minimal as to what the histo assistant should and should not do so our lab manager would like to see what other hospitals are doing and requiring for this job position. Thanks in advance, amy NOTE: The information contained in this message may be privileged, 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 this message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JeffusBrandon <@t> uams.edu Tue Sep 14 11:33:48 2004 From: JeffusBrandon <@t> uams.edu (Jeffus, Brandon) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] rat embryos Message-ID: <748D4F173E342D47B69FC23C87E5913C0125E7D4@EXCHANGE4.ad.uams.edu> Hello all... im rather new to immunohistochemistry so bear with me. I'm attempting to process some rat embryos for staining with a primary Ab and then a Cy3 labeled secondary for visualization of protein localization. I'm wondering if the best processing method would be frozen sections of the samples or paraffin embedding the samples? Any information would be greatly appreciated. Brandon C Jeffus Graduate Student From bwhitaker <@t> brownpathology.com Tue Sep 14 11:40:24 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:24:02 2005 Subject: FW: [Histonet] Histo. Assistant Job Descriptions In-Reply-To: Message-ID: <000001c49a79$88e006c0$3601a8c0@brownpathology.net> OOPS... attachments bad!! Pasting good. Below is our aid job description. Bonnie Whitaker Job Description for Laboratory/Grossing Aid Reports To: Laboratory Manager Pathologists Qualifications: Education: high school student or have diploma or GED Experience: not required Certification: not required Personal: The Lab/Grossing Aid must demonstrate an ability to listen carefully and follow directions well. The incumbent must quickly grasp the work-flow of specimen accessioning and grossing and become familiar with these procedures. The incumbent must demonstrate reasonable intelligence, diligence, accuracy, cooperation, neatness, and promptness in keeping with the prestige and image of this laboratory. The incumbent takes guidance and instruction from the pathologists, with whom they are working, as well as the Laboratory Manager and Executive Director. Responsibilities: The Lab/Grossing Aid will be responsible for the following duties: 1. Accessioning specimens and assisting with grossing procedures including: cassette labeling, pulling specimens for additional tissue submission, and discarding specimens in accordance with policy/procedure manual. 2. Assisting with the performance of routine maintenance and QC procedures on equipment to include but not limited to cryostat, grossing area, stain setup and filters. 3. Maintenance of inventory 4. Routine changing of stains with appropriate records maintenance. 5. Cleaning of the grossing area and instruments. 6. Any filing or routine clerical duties necessary. 7. Other duties as assigned. General: 1. The Lab/Grossing Aid shall be responsible for maintaining all company records of a proprietary or confidential nature secure and confidential and shall return all lists of clients, potential clients, and all other records, at Brown & Associates request. 2. The Lab/Grossing Aid is responsible for representing B&AML in a professional and dignified manner befitting the status of the laboratory in the Houston Medical Community. 3. The Lab/Grossing Aid will be expected to comply with all policies of B&AML as stated in the employee handbook and any newly instituted policies not specifically stated in the handbook. Will be expected to keep assigned work area as neat and clean as possible so as to afford the maximum functional usage of the assigned area. Will be expected to keep all records and files available and accessible to authorized personnel at all times. 4. Lab/Grossing Aid must be in reasonable good health with no record of chronic illness resulting in undue absence from work. I, the undersigned, have carefully read the above job description and fully understand all that is stated herein. I agree to perform all of the duties above as well as any other duties deemed necessary by my supervisor within the limits described here. _________________________________________ _____________________ EMPLOYEE SIGNATURE DATE _________________________________________ _____________________ LABORATORY MANAGER DATE Bonnie Whitaker -----Original Message----- From: Nita Searcy [mailto:NSEARCY@swmail.sw.org] Sent: Tuesday, September 14, 2004 11:30 AM To: bwhitaker@brownpathology.com Subject: Re: FW: [Histonet] Histo. Assistant Job Descriptions Bonnie, I can't view your description- could you fax? 254-724-4391 Thanks! >>> "Bonnie Whitaker" 9/14/2004 10:54:03 AM >>> Here is ours. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kari Bradshaw Sent: Tuesday, September 14, 2004 9:41 AM To: Amy Self; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histo. Assistant Job Descriptions Here is our job description. :) Kari Bradshaw, HT(ASCP) Laboratory Manager Lower Columbia Pathologists 1217 14th Ave Longview, WA 98632 (360)425-5620 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amy Self Sent: Tuesday, September 14, 2004 7:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histo. Assistant Job Descriptions Good Morning Netters, We are looking to hire a histology assistant in our lab. I was wandering if anyone had any job descriptions as well as job requirements that they could share with me? We have one but it very minimal as to what the histo assistant should and should not do so our lab manager would like to see what other hospitals are doing and requiring for this job position. Thanks in advance, amy NOTE: The information contained in this message may be privileged, 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 this message and deleting it from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ssalesky <@t> LowellGeneral.org Tue Sep 14 12:38:59 2004 From: ssalesky <@t> LowellGeneral.org (Shawn Salesky) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] RE: Histonet Digest, Vol 10, Issue 19 Message-ID: <9E49B4E3A6200C4B91E5FB99FB1F7EDB571F4E@NTMAIL> Hello All, The Lab I work in is currently setting up automated IHC, ISH and Special stains on Ventana systems. First question... Is anyone using Ventana Inform HPV HR in a clinical setting? and How did you validate? Second question... Is there a standard number of cases to run in validations of Immuno's? Third question... What numbers did you use to validate your special stains? I'm pretty much at a loss, as I cannot find any literature on the subjects. Thanks in advance, Shawn Salesky TC LGH Histo > ---------- > From: > histonet-bounces@lists.utsouthwestern.edu[SMTP:histonet-bounces@lists.utso > uthwestern.edu] on behalf of > histonet-request@lists.utsouthwestern.edu[SMTP:histonet-request@lists.utso > uthwestern.edu] > Sent: Tuesday, September 14, 2004 1:08 PM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet Digest, Vol 10, Issue 19 > > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. Re: Anti-parrafin floor (DDittus787@aol.com) > 2. re embedding of small artery (carl hobbs) > 3. Recover tissue sections on non-plus slides (Kim Byrnes) > 4. RE: Recover tissue sections on non-plus slides (Bonnie Whitaker) > 5. Re: Recover tissue sections on non-plus slides > (chiggerson@memhosp.com) > 6. alizarin red fading (Julien) > 7. fluorescent dyes/lables for neuronal activities? (Cao, Xudong) > 8. RE: alizarin red fading (Barry R Rittman) > 9. RE: Recover tissue sections on non-plus slides (Laurie Colbert) > 10. cryomacrocut (Robin Turcotte) > 11. cryomacrocut (Robin Turcotte) > 12. cryomacrocut (Robin Turcotte) > 13. cryomacrocut (Robin Turcotte) > 14. cryomacrocut (Robin Turcotte) > 15. Shandon Hypercenter Xp (Jim Staruk) > 16. Superfrost Slides (Bill Sinai) > 17. embedding of small artery_paraffin (Dr. Raghul) > 18. vascular graft_sectioning difficulty (Dr. Raghul) > 19. Test (Dimaano, Nena) > 20. RE: Superfrost Slides (Joe Nocito) > 21. Perferred fixative for animals > (wasielewski.reinhard.von@mh-hannover.de) > 22. RE: Fixative for animals (Cullen, Kay) > 23. Histo. Assistant Job Descriptions (Amy Self) > 24. Re: Perferred fixative for animals (LaCinda Burchell) > 25. Re: Recover tissue sections on non-plus slides (Barbara Lentz) > 26. RE: Histo. Assistant Job Descriptions (Kari Bradshaw) > 27. FW: [Histonet] Histo. Assistant Job Descriptions (Bonnie Whitaker) > 28. rat embryos (Jeffus, Brandon) > 29. RE: FW: [Histonet] Histo. Assistant Job Descriptions > (Bonnie Whitaker) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Mon, 13 Sep 2004 13:48:40 -0400 > From: DDittus787@aol.com > Subject: [Histonet] Re: Anti-parrafin floor > To: BBEIER@kumc.edu ("Barbara Beier") > Cc: histonet@pathology.swmed.edu > Message-ID: <7B0A3F7A.56A40CDC.0A1F969F@aol.com> > Content-Type: text/plain; charset=iso-8859-1 > > Barb: > the flooring came from Acme Panels-the company is in the UK and can be > found on Google, it is anti-solvent, chemical, paraffin,etc. It comes in 3 > or 4 colors and everyone loves it. > We had a contractor bring it in and install it. > dana > > > > ------------------------------ > > Message: 2 > Date: Mon, 13 Sep 2004 19:02:14 +0100 > From: "carl hobbs" > Subject: [Histonet] re embedding of small artery > To: "Histonet" > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > In my experienc, with fixed tissues it is easy to embed using agarose, > then > to re-orient the tissue for frozen/pwax sectioning: Dissolve 1% agarose( > aq) > and when about 50C place the tissue in the agarose, in a bigger mould. It > will set quickly.....better if placed at 4C for a while. Remove the set > agarose and cut away excess agarose so you have the tissue in the > orientation of your choice. Then process to pwax. Or, place in OCT for a > couple of hours on a rotator. Then embed in an OCT filled mould in the > orientation you need and....freeze. If you are doing fresh tissue, get the > melted, cooling agarose into a mould , place the tissue and cool quickly. > Immediately place in OCT, swirl gently for a minute, then freeze. Play > around with control material until you are confident of the technique and > to > reassure yourself that the elevated temp does not adversely afffect your > tissue. You can also do gelatin/vibratome sections of the fixed tissue, if > you have a vibratome. > --- > Outgoing mail is certified Virus Free. > Checked by AVG anti-virus system (http://www.grisoft.com). > Version: 6.0.754 / Virus Database: 504 - Release Date: 06/09/2004 > > > > > ------------------------------ > > Message: 3 > Date: Mon, 13 Sep 2004 11:34:29 -0700 (PDT) > From: Kim Byrnes > Subject: [Histonet] Recover tissue sections on non-plus slides > To: histonet@lists.utsouthwestern.edu > Message-ID: <20040913183429.84652.qmail@web12308.mail.yahoo.com> > Content-Type: text/plain; charset=us-ascii > > Does anyone have a method or know of a product that > can recover or rescue tissue sections that were > accidentally mounted onto non-plus (non-polarized, > non-coated) slides? We would like to do immuno on > these slides, but they consistently fall off of the > slides. > > Any ideas? > > Thanks, > > Kim > Georgetown University > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - 50x more storage than other providers! > http://promotions.yahoo.com/new_mail > > > > ------------------------------ > > Message: 4 > Date: Mon, 13 Sep 2004 14:24:14 -0500 > From: "Bonnie Whitaker" > Subject: RE: [Histonet] Recover tissue sections on non-plus slides > To: "'Kim Byrnes'" > Cc: histonet@lists.utsouthwestern.edu > Message-ID: <000301c499c7$41758920$3601a8c0@brownpathology.net> > Content-Type: text/plain; charset="us-ascii" > > There is a procedure to remove tissue sections from a slide, and they can > then be mounted on + slides. The product that works best in my hands is > Mount-Quick. It has more than one distributer, but I know that Newcomer > Supply (800-383-7799) has this product, as well as directions on how to do > the procedure. > > Bonnie Whitaker > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Byrnes > Sent: Monday, September 13, 2004 1:34 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Recover tissue sections on non-plus slides > > Does anyone have a method or know of a product that > can recover or rescue tissue sections that were > accidentally mounted onto non-plus (non-polarized, > non-coated) slides? We would like to do immuno on > these slides, but they consistently fall off of the > slides. > > Any ideas? > > Thanks, > > Kim > Georgetown University > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - 50x more storage than other providers! > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 5 > Date: Mon, 13 Sep 2004 14:46:54 -0500 > From: chiggerson@memhosp.com > Subject: Re: [Histonet] Recover tissue sections on non-plus slides > To: histonet@lists.utsouthwestern.edu > Cc: Kim Byrnes > Message-ID: > > Content-Type: text/plain; charset="US-ASCII" > > Kim, > > The 2000 Tech Sample exercise HT-1 gives a procedure for this. The title > of the exercise is "Use Mount Quick to Remove Tissue Sections from > Non-Positively Charged Slides for Immunohistochemical and Special Stains". > > > Good luck! > > Cindy > > Cindy Higgerson HTL(ASCP) > Surgical Pathology Supervisor > Memorial Hospital > > > > > > Kim Byrnes > Sent by: histonet-bounces@lists.utsouthwestern.edu > 09/13/2004 01:34 PM > > To > histonet@lists.utsouthwestern.edu > cc > > Subject > [Histonet] Recover tissue sections on non-plus slides > > > > > > > Does anyone have a method or know of a product that > can recover or rescue tissue sections that were > accidentally mounted onto non-plus (non-polarized, > non-coated) slides? We would like to do immuno on > these slides, but they consistently fall off of the > slides. > > Any ideas? > > Thanks, > > Kim > Georgetown University > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - 50x more storage than other providers! > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 6 > Date: Mon, 13 Sep 2004 16:58:59 -0400 > From: "Julien" > Subject: [Histonet] alizarin red fading > To: > Message-ID: <00c501c499d4$7dc51140$9310d784@uqar.qc.ca> > Content-Type: text/plain; charset="iso-8859-1" > > Hello all, > > We do differencial staining of bone and cartillage on small fish (~1cm). > To do so, we use the Clear and Stain technique which results in red bone > coloration and blue cartillage coloration. After a month or so, We noticed > that the red stain tends to fade off. We tried to re-stain the fish, but > the alizarin red wouldn't stain the bones anymore. > > Why is this so? Is it that bone decalcifies in the glycerin? We put a few > cristals of thymol in the glycerin to avoid mold development. Could this > be a decalcifying agent? Does anybody know of a way to re-stain? > > For the time being, we score bony elements within 3 days of staining to > avoid fading problems, but this stops us from doing batch staining. Batch > staining would be much faster. We would then keep stained fish in glycerin > until scoring. > > Thanks for any suggestion. > > Julien Lambrey de Souza > Biologie ?volutive, > Universit? du Qu?bec ? Rimouski, > > > Julien Lambrey de Souza > Biologie ?volutive, > Universit? du Qu?bec ? Rimouski, > > (418) 723-1986 #1438 > > > ------------------------------ > > Message: 7 > Date: Mon, 13 Sep 2004 17:03:50 -0400 > From: "Cao, Xudong" > Subject: [Histonet] fluorescent dyes/lables for neuronal activities? > To: > Message-ID: > <1DA88DDC48CCC245A777599FDAEED6D0A3909B@MAIL1.AD.Brown.Edu> > Content-Type: text/plain; charset="iso-8859-1" > > Greetings! first of all I'd like to thank eveybody who responded to my > previous posts. The advice/ideas I received were valuable! Here I have > one more question: I am studying nerve innervation into artificial skin > graft that we make in vitro and my immunostaining tells me that there are > some wonderful nerve fibers growing into the skin. The question that we > have now is if the innervation functional. In order to test this, we want > to mechanically tease the artificial skin and see if there is some > responses from the nerve cells that have presumablly innervated the > corresponding part of the skin. Thus, it would be nice to find some > fluorescent dyes/lables that will glow when the neurons are > activated/firing. any suggestions? > > best regards, > > Xudong > > > > ------------------------------ > > Message: 8 > Date: Mon, 13 Sep 2004 16:29:32 -0500 > From: "Barry R Rittman" > Subject: RE: [Histonet] alizarin red fading > To: "histonet" > Message-ID: > <566FB0B522443D43AF02D2ADBE35A6F0F13357@UTHEVS3.mail.uthouston.edu> > Content-Type: text/plain; charset="iso-8859-1" > > I cannot see why the addition of a few crystals of thymol would make the > glycerin acidic enough to cause removal of calcium and thus leaching of > the alizarin. > If you are using 100% glycerin, there should be no need for the addition > of thymol. > I am not sure exactly how you have processed these specimens. If you are > using the classical method with potassium hydroxide (as the leaching agent > to remove excess stain from soft tissues), is it possible that there were > traces of potassium hydroxide left in the specimen? > I have some specimens that were stained in 1961 and some in 1970 that show > no signs of fading. > Barry > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Julien > Sent: Monday, September 13, 2004 3:59 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] alizarin red fading > > Hello all, > > We do differencial staining of bone and cartillage on small fish (~1cm). > To do so, we use the Clear and Stain technique which results in red bone > coloration and blue cartillage coloration. After a month or so, We noticed > that the red stain tends to fade off. We tried to re-stain the fish, but > the alizarin red wouldn't stain the bones anymore. > > Why is this so? Is it that bone decalcifies in the glycerin? We put a few > cristals of thymol in the glycerin to avoid mold development. Could this > be a decalcifying agent? Does anybody know of a way to re-stain? > > For the time being, we score bony elements within 3 days of staining to > avoid fading problems, but this stops us from doing batch staining. Batch > staining would be much faster. We would then keep stained fish in glycerin > until scoring. > > Thanks for any suggestion. > > Julien Lambrey de Souza > Biologie ?volutive, > Universit? du Qu?bec ? Rimouski, > > > Julien Lambrey de Souza > Biologie ?volutive, > Universit? du Qu?bec ? Rimouski, > > (418) 723-1986 #1438 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 9 > Date: Mon, 13 Sep 2004 14:43:49 -0700 > From: "Laurie Colbert" > Subject: RE: [Histonet] Recover tissue sections on non-plus slides > To: "Kim Byrnes" , "Histonet (E-mail)" > > Message-ID: > > <0BE6ADFAE4E7E04496BF21ABD346628001C5C062@EXCHANGE1.huntingtonhospital.com > > > > Content-Type: text/plain; charset="iso-8859-1" > > We have used the Mount Quick from Newcomer Supply, and it works well. > > Laurie Colbert > Huntington Memorial Hospital > Pasadena, CA > > -----Original Message----- > From: Kim Byrnes [mailto:k_byrnes@yahoo.com] > Sent: Monday, September 13, 2004 11:34 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Recover tissue sections on non-plus slides > > > Does anyone have a method or know of a product that > can recover or rescue tissue sections that were > accidentally mounted onto non-plus (non-polarized, > non-coated) slides? We would like to do immuno on > these slides, but they consistently fall off of the > slides. > > Any ideas? > > Thanks, > > Kim > Georgetown University > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - 50x more storage than other providers! > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 10 > Date: Mon, 13 Sep 2004 20:01:58 -0400 > From: "Robin Turcotte" > Subject: [Histonet] cryomacrocut > To: > Message-ID: <000701c499ee$cb688f70$df8ceccf@client> > Content-Type: text/plain; charset="iso-8859-1" > > Does anyone have an address from a compagny or an hospital who have a used > cryomacrocut in good condition to be sold. > > Thank you > > Robin > > > > > ------------------------------ > > Message: 11 > Date: Mon, 13 Sep 2004 20:07:19 -0400 > From: "Robin Turcotte" > Subject: [Histonet] cryomacrocut > To: > Message-ID: <000e01c499ee$ee9cdbe0$df8ceccf@client> > Content-Type: text/plain; charset="iso-8859-1" > > Does anyone have an address from a compagny or an hospital who have a used > cryomacrocut in good condition to be sold. > > Thank you > > Robin > > > > > ------------------------------ > > Message: 12 > Date: Mon, 13 Sep 2004 20:07:47 -0400 > From: "Robin Turcotte" > Subject: [Histonet] cryomacrocut > To: > Message-ID: <000f01c499ee$ef064c10$df8ceccf@client> > Content-Type: text/plain; charset="iso-8859-1" > > Does anyone have an address from a compagny or an hospital who have a used > cryomacrocut in good condition to be sold. > > Thank you > > Robin > > > > > ------------------------------ > > Message: 13 > Date: Mon, 13 Sep 2004 20:10:32 -0400 > From: "Robin Turcotte" > Subject: [Histonet] cryomacrocut > To: > Message-ID: <001301c499ef$45100dd0$df8ceccf@client> > Content-Type: text/plain; charset="iso-8859-1" > > Does anyone have an address from a compagny or an hospital who have a used > cryomacrocut in good condition to be sold. > > Thank you > > Robin > > > > > ------------------------------ > > Message: 14 > Date: Mon, 13 Sep 2004 20:11:36 -0400 > From: "Robin Turcotte" > Subject: [Histonet] cryomacrocut > To: > Message-ID: <001c01c499ef$6fce96e0$df8ceccf@client> > Content-Type: text/plain; charset="iso-8859-1" > > Does anyone have an address from a compagny or an hospital who have a used > cryomacrocut in good condition to be sold. > > Thank you > > Robin > > > > ------------------------------ > > Message: 15 > Date: Mon, 13 Sep 2004 21:15:52 -0400 > From: "Jim Staruk" > Subject: [Histonet] Shandon Hypercenter Xp > To: > Message-ID: <000201c499f8$87d3cb80$6401a8c0@yourw04gtxld67> > Content-Type: text/plain; charset="us-ascii" > > I'm looking for someone familiar with the Shandon Hypercenter Xp tissue > processor. I need some technical advise. Can someone help me? Anyone > planning to invoice me for their help need not reply. > > Thank you > > Jim > > ________________________ > James E. Staruk, HT(ASCP) > Mass Histopathology Service > www.masshistology.com > > > > > > ------------------------------ > > Message: 16 > Date: Tue, 14 Sep 2004 13:43:17 +1000 > From: "Bill Sinai" > Subject: [Histonet] Superfrost Slides > To: "histonet \(E-mail\)" > Message-ID: <000101c49a0c$f9356e50$e1ce080a@wsahs.nsw.gov.au> > Content-Type: text/plain; charset="iso-8859-1" > > > Is anyone having troubles with NO staining on some slides on the Ventana > Benchmark. > The biggest problem seems to be with the EDTA buffered CC1 retrieval > solution. > Not all slides are effected, just an occasional one, but we usually end up > with little or no staining with anything including the Haematoxylin. > We use Menzel Superforst Plus and Superfrost Plus Ultra, and both slides > have shown the effect. > > Thanks > Bill Sinai > Laboratory Manager > Tissue Pathology, ICPMR > Westmead NSW 2145 > Australia > Ph 02 9845 7774 > > > __________________________________________________________________ > > This electronic message and any attachments may be confidential. If you > are not the intended recipient of this message would you please delete the > message and any attachments and advise the sender. 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. > > > > ------------------------------ > > Message: 17 > Date: Tue, 14 Sep 2004 16:31:15 +0530 > From: "Dr. Raghul" > Subject: [Histonet] embedding of small artery_paraffin > To: histonet@lists.utsouthwestern.edu > Message-ID: > > hello galina, > > Rat and mouse organs can be cut easily after short processing(using > acetone and xylene)and then embedding in paraffin wax of low melting point > (56 C). We have followed the protocols from below given reference > successfully and same could applied for the small artery also > > Theory and practice of Histological techniques (bancroft and steven) > > (or) > > Manual of histologic staining methods of the armed forces institutes of > pathology edited by Lee G. Luna, III edition page 16 > > > Raghul J > Histopathology-Implant Biology > Sree chitra tirunal institute > Thiruvananthapuram 695 012 > > > > > ------------------------------ > > Message: 18 > Date: Tue, 14 Sep 2004 16:56:23 +0530 > From: "Dr. Raghul" > Subject: [Histonet] vascular graft_sectioning difficulty > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > hello members, > > We cut 5 ?m thin sections of paraffin embedded vascular graft(Dacron) in > a rotary microtome with disposable blades but the senior tech tells that > the tissue sections are crumbling and forces us to go for thick sections > around 10?m but again not with much success. > > We can understand that there is not enough tissue adhereing to the > graft and we are literally cutting the graft. But did anyone had such > problems sectioning vascular grafts. Any suggestions? > > > regards , > > Raghul J > scientist > Histopathology- implant biology division > Srichitra tirunal institute of medical sciences and technology > Trivandrum -695 012 > Kerala, India > > > > > ------------------------------ > > Message: 19 > Date: Tue, 14 Sep 2004 07:54:55 -0400 > From: "Dimaano, Nena" > Subject: [Histonet] Test > To: > Message-ID: > > <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD06210EE4@HOS2KEXCHCL.howost.strykercorp.c > om> > > Content-Type: text/plain; charset="iso-8859-1" > > > Nena Dimaano > Advanced Technology > Stryker Orthopaedics > 325 Corporate Drive > Mahwah, NJ 07430 > tel: 201-831-5338 > fax: 201-831-6224 > email: Nena.Dimaano@stryker.com > > > > ------------------------------ > > Message: 20 > Date: Tue, 14 Sep 2004 07:09:09 -0500 > From: "Joe Nocito" > Subject: RE: [Histonet] Superfrost Slides > To: "Bill Sinai" , "histonet > \(E-mail\)" > > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > Bill, > we've had problems also. It could be that the slides might be too changed, > which is causing the reagents to bead up. Currently, we are using positive > charged slides from Statlab Medical Products, which I think they get from > Erie Scientific. Statlab's number is 1-800-442-3573. The may be able to > direct you to some one closer to you. > > 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 Bill > Sinai > Sent: Monday, September 13, 2004 10:43 PM > To: histonet (E-mail) > Subject: [Histonet] Superfrost Slides > > > > Is anyone having troubles with NO staining on some slides on the Ventana > Benchmark. > The biggest problem seems to be with the EDTA buffered CC1 retrieval > solution. > Not all slides are effected, just an occasional one, but we usually end up > with little or no staining with anything including the Haematoxylin. > We use Menzel Superforst Plus and Superfrost Plus Ultra, and both slides > have shown the effect. > > Thanks > Bill Sinai > Laboratory Manager > Tissue Pathology, ICPMR > Westmead NSW 2145 > Australia > Ph 02 9845 7774 > > > __________________________________________________________________ > > This electronic message and any attachments may be confidential. If you > are not the intended recipient of this message would you please delete the > message and any attachments and advise the sender. 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. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 21 > Date: Tue, 14 Sep 2004 14:29:10 +0200 > From: wasielewski.reinhard.von@mh-hannover.de > Subject: [Histonet] Perferred fixative for animals > To: histonet@lists.utsouthwestern.edu > Message-ID: <41470036.30954.8F79EDC3@localhost> > Content-Type: text/plain; charset=US-ASCII > > Hi Histonetters, > We are looking for the most convenient and morpholgical satisfying > fixative for > mouse and rat tissue beside formaldehyde. We want to stain a wide range of > > different immunomarkers but don' t like formalin (therefore) too much. > Frozen > sections are too bad in morphology. > Please send me any suggestions ! > > Many thanks in advance, > Reinhard. > > > > PD Dr. med. Reinhard von Wasielewski > > > > > ------------------------------ > > Message: 22 > Date: Tue, 14 Sep 2004 08:56:22 -0400 > From: "Cullen, Kay" > Subject: [Histonet] RE: Fixative for animals > To: "Histonet \(E-mail\)" > Message-ID: > > > Content-Type: text/plain; charset="iso-8859-1" > > Are you busing straight formaldehyde or formaldehyde/glutaraldehyde? I > get very good results with Bouin's for most tissue. > > Kay Cullen > Research Associate > Department of Cell Biology > University of Massachusetts Medical School > Worcester, MA USA > > > > ------------------------------ > > Message: 23 > Date: Tue, 14 Sep 2004 10:05:39 -0400 > From: "Amy Self" > Subject: [Histonet] Histo. Assistant Job Descriptions > To: > Message-ID: > <39836CD6DB61654E8F95A35898C921860A87F3@exchange.gmhpost.com> > Content-Type: text/plain; charset="iso-8859-1" > > > > Good Morning Netters, > > We are looking to hire a histology assistant in our lab. I was > wandering if anyone had any job descriptions as well as job requirements > that they could share with me? We have one but it very minimal as to what > the histo assistant should and should not do so our lab manager would like > to see what other hospitals are doing and requiring for this job position. > Thanks in advance, amy > > > > > > > NOTE: > The information contained in this message may be privileged, 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 this message and deleting it from > your computer. Thank you. > > > ------------------------------ > > Message: 24 > Date: Tue, 14 Sep 2004 09:16:54 -0500 > From: "LaCinda Burchell" > Subject: Re: [Histonet] Perferred fixative for animals > To: , > > Message-ID: > Content-Type: text/plain; charset=US-ASCII > > Dear Reinhard, > We use Prefer fixative from Anatech Ltd, www.anatechltdusa. We have > beautiful morphology, staining is wonderful (specials and immunos). The > only down side of Prefer is that it affects the staining of eosinophils. > We've recently learned how to get around that issue in human tissues, > but haven't figured it out in rat tissues. Best of luck! Cindy B > > LaCinda Burchell, BA, AS, HT(ASCP) > University of Wisconsin-Madison, Medical School > Asthma and Allergy Research IHC Lab > 600 Highland Ave. CSC K4/913 > Madison, Wisconsin 53792 > > Phone: 608-262-3518 > FAX: 608-263-3746 > > >>> 09/14/04 07:29AM >>> > Hi Histonetters, > We are looking for the most convenient and morpholgical satisfying > fixative for > mouse and rat tissue beside formaldehyde. We want to stain a wide range > of > different immunomarkers but don' t like formalin (therefore) too much. > Frozen > sections are too bad in morphology. > Please send me any suggestions ! > > Many thanks in advance, > Reinhard. > > > > PD Dr. med. Reinhard von Wasielewski > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 25 > Date: Tue, 14 Sep 2004 10:48:20 -0400 > From: "Barbara Lentz" > Subject: Re: [Histonet] Recover tissue sections on non-plus slides > To: , > Message-ID: > Content-Type: text/plain; charset=US-ASCII > > We, too, use the Mount Quick from Newcomer Supply. On a regular basis, > we remove H&E tissue sections to perform immunos. > www.newcomersupply.com is their address, Barb > > >>> Kim Byrnes 09/13/04 02:34PM >>> > Does anyone have a method or know of a product that > can recover or rescue tissue sections that were > accidentally mounted onto non-plus (non-polarized, > non-coated) slides? We would like to do immuno on > these slides, but they consistently fall off of the > slides. > > Any ideas? > > Thanks, > > Kim > Georgetown University > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - 50x more storage than other providers! > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 26 > Date: Tue, 14 Sep 2004 07:41:17 -0700 > From: "Kari Bradshaw" > Subject: RE: [Histonet] Histo. Assistant Job Descriptions > To: "Amy Self" , > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > Here is our job description. > > :) Kari Bradshaw, HT(ASCP) > Laboratory Manager > Lower Columbia Pathologists > 1217 14th Ave > Longview, WA 98632 > (360)425-5620 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amy Self > Sent: Tuesday, September 14, 2004 7:06 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histo. Assistant Job Descriptions > > > > > Good Morning Netters, > > We are looking to hire a histology assistant in our lab. I was > wandering > if anyone had any job descriptions as well as job requirements that they > could share with me? We have one but it very minimal as to what the histo > assistant should and should not do so our lab manager would like to see > what > other hospitals are doing and requiring for this job position. Thanks in > advance, amy > > > > > > > NOTE: > The information contained in this message may be privileged, 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 this message and deleting it from your > computer. Thank you. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ > > Message: 27 > Date: Tue, 14 Sep 2004 10:54:03 -0500 > From: "Bonnie Whitaker" > Subject: FW: [Histonet] Histo. Assistant Job Descriptions > To: > Message-ID: <000801c49a73$0ee10140$3601a8c0@brownpathology.net> > Content-Type: text/plain; charset="us-ascii" > > Here is ours. > Bonnie Whitaker > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kari > Bradshaw > Sent: Tuesday, September 14, 2004 9:41 AM > To: Amy Self; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histo. Assistant Job Descriptions > > Here is our job description. > > :) Kari Bradshaw, HT(ASCP) > Laboratory Manager > Lower Columbia Pathologists > 1217 14th Ave > Longview, WA 98632 > (360)425-5620 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amy Self > Sent: Tuesday, September 14, 2004 7:06 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histo. Assistant Job Descriptions > > > > > Good Morning Netters, > > We are looking to hire a histology assistant in our lab. I was > wandering > if anyone had any job descriptions as well as job requirements that they > could share with me? We have one but it very minimal as to what the histo > assistant should and should not do so our lab manager would like to see > what > other hospitals are doing and requiring for this job position. Thanks in > advance, amy > > > > > > > NOTE: > The information contained in this message may be privileged, 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 this message and deleting it from your > computer. Thank you. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ > > Message: 28 > Date: Tue, 14 Sep 2004 11:33:48 -0500 > From: "Jeffus, Brandon" > Subject: [Histonet] rat embryos > To: > Message-ID: > <748D4F173E342D47B69FC23C87E5913C0125E7D4@EXCHANGE4.ad.uams.edu> > Content-Type: text/plain; charset="us-ascii" > > Hello all... im rather new to immunohistochemistry so bear with me. I'm > attempting to process some rat embryos for staining with a primary Ab > and then a Cy3 labeled secondary for visualization of protein > localization. I'm wondering if the best processing method would be > frozen sections of the samples or paraffin embedding the samples? Any > information would be greatly appreciated. > > > > Brandon C Jeffus > > Graduate Student > > > > ------------------------------ > > Message: 29 > Date: Tue, 14 Sep 2004 11:40:24 -0500 > From: "Bonnie Whitaker" > Subject: RE: FW: [Histonet] Histo. Assistant Job Descriptions > To: "'Nita Searcy'" > Cc: histonet@lists.utsouthwestern.edu > Message-ID: <000001c49a79$88e006c0$3601a8c0@brownpathology.net> > Content-Type: text/plain; charset="us-ascii" > > > OOPS... attachments bad!! Pasting good. > > > Below is our aid job description. > > Bonnie Whitaker > > > Job Description for Laboratory/Grossing Aid > > > > > Reports To: Laboratory Manager > > Pathologists > > > > Qualifications: > > > > Education: high school student or have > diploma > or GED > > > > Experience: not required > > > > Certification: not required > > > > Personal: The Lab/Grossing Aid must > demonstrate an ability to listen carefully and follow directions well. > The > incumbent must quickly grasp the work-flow of specimen accessioning and > grossing and become familiar with these procedures. The incumbent must > demonstrate reasonable intelligence, diligence, accuracy, cooperation, > neatness, and promptness in keeping with the prestige and image of this > laboratory. The incumbent takes guidance and instruction from the > pathologists, with whom they are working, as well as the Laboratory > Manager > and Executive Director. > > Responsibilities: The Lab/Grossing Aid will be > responsible for the following duties: > > 1. Accessioning specimens and assisting with grossing procedures > including: cassette labeling, pulling specimens for additional tissue > submission, and discarding specimens in accordance with policy/procedure > manual. > > 2. Assisting with the performance of routine maintenance and QC > procedures on equipment to include but not limited to cryostat, grossing > area, stain setup and filters. > > 3. Maintenance of inventory > > 4. Routine changing of stains with appropriate records maintenance. > > 5. Cleaning of the grossing area and instruments. > > 6. Any filing or routine clerical duties necessary. > > 7. Other duties as assigned. > > > > > General: > > 1. The Lab/Grossing Aid shall be responsible for maintaining all > company > records of a proprietary or confidential nature secure and confidential > and > shall return all lists of clients, potential clients, and all other > records, > at Brown & Associates request. > > 2. The Lab/Grossing Aid is responsible for representing B&AML in a > professional and dignified manner befitting the status of the laboratory > in > the Houston Medical Community. > > 3. The Lab/Grossing Aid will be expected to comply with all policies > of > B&AML as stated in the employee handbook and any newly instituted policies > not specifically stated in the handbook. Will be expected to keep > assigned > work area as neat and clean as possible so as to afford the maximum > functional usage of the assigned area. Will be expected to keep all > records > and files available and accessible to authorized personnel at all times. > > 4. Lab/Grossing Aid must be in reasonable good health with no record > of > chronic illness resulting in undue absence from work. > > > > I, the undersigned, have carefully read the above job description and > fully > understand all that is stated herein. I agree to perform all of the > duties > above as well as any other duties deemed necessary by my supervisor within > the limits described here. > > > > > > _________________________________________ > _____________________ > > > EMPLOYEE SIGNATURE > DATE > > > > > > > > _________________________________________ > _____________________ > > LABORATORY MANAGER DATE > > > > > > > > > > Bonnie Whitaker > > -----Original Message----- > From: Nita Searcy [mailto:NSEARCY@swmail.sw.org] > Sent: Tuesday, September 14, 2004 11:30 AM > To: bwhitaker@brownpathology.com > Subject: Re: FW: [Histonet] Histo. Assistant Job Descriptions > > > > Bonnie, I can't view your description- could you fax? 254-724-4391 > > Thanks! > > >>> "Bonnie Whitaker" 9/14/2004 10:54:03 AM > >>> > > Here is ours. > Bonnie Whitaker > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Kari > Bradshaw > Sent: Tuesday, September 14, 2004 9:41 AM > To: Amy Self; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histo. Assistant Job Descriptions > > Here is our job description. > > :) Kari Bradshaw, HT(ASCP) > Laboratory Manager > Lower Columbia Pathologists > 1217 14th Ave > Longview, WA 98632 > (360)425-5620 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On > Behalf Of Amy > Self > Sent: Tuesday, September 14, 2004 7:06 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histo. Assistant Job Descriptions > > > > > Good Morning Netters, > > We are looking to hire a histology assistant in our lab. I was > wandering > if anyone had any job descriptions as well as job requirements that they > could share with me? We have one but it very minimal as to what the histo > assistant should and should not do so our lab manager would like to see > what > other hospitals are doing and requiring for this job position. Thanks in > advance, amy > > > > > > > NOTE: > The information contained in this message may be privileged, 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 this 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 > > End of Histonet Digest, Vol 10, Issue 19 > **************************************** > This e-mail and any attachments is intended only for the person or entity to which it is addressed and may contain confidential and privileged information. If you are not the intended recipient, please notify the sender immediately by return e-mail, delete this e-mail and destroy any copies. Any dissemination or use of this information by a person other than the intended recipient is unauthorized and may be illegal. From Michele_Marggi <@t> ssmhc.com Tue Sep 14 12:42:44 2004 From: Michele_Marggi <@t> ssmhc.com (Michele_Marggi@ssmhc.com) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Coverslipping- Glass vs. Film/Tape Message-ID: For all those having problems with the tissue and film coming off of the slide after an extended period of time, can you tell me what type of 1) coverslipper you are using and 2) what type of film/tape? I want to know if I should be closely monitoring our older slides, but I understand that the type (brand) of tape may be the determining factor. We have been using film for 10 years now, and don't have any problems to date, and obviously want to keep it that way. Michele Marggi Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 From georgecole <@t> ev1.net Tue Sep 14 12:59:51 2004 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] fixed and unfixed immuno and antuibody oreps Message-ID: <000601c49a84$a5c46a50$064dbad0@hppav> I promise to drop this line---I have had an itch to find out something--- I realize I must give it up---I have asked it repeatedly and I don't want to keep asking the same thing---impatience is bound to develop. Retired as I am---I no longer have lab in which to answer such questions. Most histotechs nowadays are brought up doing one side of the question---indeed it is the standard procedure and my question just doesn't scan with today's techs. But if I could shuck 10 years and be back in my lab and I was given the responsibility to do a certain class of work I would do the following: Antibody technique or immunofluorescence procedure---any procedure that has a fixative in its beginning followed by some procedure to neutralize the effects of the fixative----I would run at least 3 trials on the 2 sets of pairs: 1) Tissue unfixed 2) tissue fixed, then processed with by whatever step was supposed to neutralize the fixation effect, rinsing the tissues carefully afterwards. . Then I would run tissues 1 and 2 in the same preparation. And I would choose as my standard procedure which ever procedure of the pair which gave the better results. I drop that and run. The crowding effect of Standard Procedure Itis is out there. But then I might just find my question was trivial and standard procedure might win. But has the fix-unfix procedure ever been tested against the unfixed prep? This retired picky-compulsive tech would never run ANY procedure as standard without testing its effectiveness---comparing it--- with any reasonable alternative procedure for quality. Nuff said. Retired Tech now enters a stage of quiet upon the subject on the histonet, but I will still brood over the untapped quality test in ANY procedure done by rote rather than via the search for the most excellent way to do that test. georgecole@ev1.net From cwscouten <@t> myneurolab.com Tue Sep 14 13:42:15 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Perferred fixative for animals Message-ID: It might not be the formaldehyde. Your freezing process sounds like a more likely culprit. Read the following article: http://www.myneurolab.com/neuronews/default.asp?AID=63&issueid=18&sitebannerid=&stylesheetid= Use formaldehyde freshly made with paraformaldehyde in buffer (formalin bought as such degrades to include acids and acetone in it), include glutaraldehype. Recipes may be seen at many places, including the manual for the Perfusion One. http://www.myneurolab.com/global/manuals/Perfusion%20One.pdf The faster the fixative gets there after start of perfusion (I assume you are using perfusion) the better. The Perfusion One can help with that. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=Sacrifice+Equipment&idsubcategory=21 Do not overfix. Do not store in fixative for over a day. 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 wasielewski.reinhard.von@mh-hannover.de Sent: Tuesday, September 14, 2004 7:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Perferred fixative for animals Hi Histonetters, We are looking for the most convenient and morpholgical satisfying fixative for mouse and rat tissue beside formaldehyde. We want to stain a wide range of different immunomarkers but don' t like formalin (therefore) too much. Frozen sections are too bad in morphology. Please send me any suggestions ! Many thanks in advance, Reinhard. PD Dr. med. Reinhard von Wasielewski _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Tue Sep 14 14:19:52 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] unsubscribing Message-ID: Hello netters, just a couple of weeks ago, instructions for unsubscribing to the Histonet were posted. Since I have no intentions of unsubscribing to the Histonet, it was suggested that since I'll be in Toronto for a week, to unsubscribe temporarily. Any one have instruction? Okay, Okay flame away. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX From JQB7 <@t> CDC.GOV Tue Sep 14 14:40:09 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] unsubscribing Message-ID: I think you have to unsubscribe completely and then resubscribe when you return. But I'm not sure. That's just what I do. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Tuesday, September 14, 2004 3:20 PM To: Histonet Subject: [Histonet] unsubscribing Hello netters, just a couple of weeks ago, instructions for unsubscribing to the Histonet were posted. Since I have no intentions of unsubscribing to the Histonet, it was suggested that since I'll be in Toronto for a week, to unsubscribe temporarily. Any one have instruction? Okay, Okay flame away. 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 kalschev <@t> svm.vetmed.wisc.edu Tue Sep 14 15:26:03 2004 From: kalschev <@t> svm.vetmed.wisc.edu (Vicki Kalscheur) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Hard Tissue Committee Meeting Message-ID: <007101c49a99$0ea95a30$65d56880@kalscheur426> Dear members and interested individuals, Please join us on Saturday, September 18th, 2004 from 5:30 - 6:30 pm in Toronto, Ontario - Canada, as we gather at NSH's 30th Annual Symposium/Convention. The room number will be posted at the Hard Tissue Display Table. I look forward to seeing many of you! Sincerely, Vicki Kalscheur, Chairperson From JWEEMS <@t> sjha.org Tue Sep 14 15:10:44 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Unsubscribing Message-ID: <83AACDB0810528418AA106F9AE9B7F7E2782C6@sjhaexc02.sjha.org> Instructions from Herb..... If you ever want to unsubscribe or change your options (eg, switch to or from digest mode, change your password, etc.), visit your subscription page at: http://lists.utsouthwestern.edu/mailman/options/histonet/jweems%40sjha.org Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From POWELL_SA <@t> Mercer.edu Tue Sep 14 15:12:47 2004 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] unsubscribing In-Reply-To: Message-ID: No you don't have to unsubscribe just go to http://lists.utsouthwestern.edu/mailman/options/histonet/, login with your password, then go to mail delivery and check disable until you return, then do the same and click enable. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bartlett, Jeanine Sent: Tuesday, September 14, 2004 2:40 PM To: Joe Nocito; Histonet Subject: RE: [Histonet] unsubscribing I think you have to unsubscribe completely and then resubscribe when you return. But I'm not sure. That's just what I do. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Tuesday, September 14, 2004 3:20 PM To: Histonet Subject: [Histonet] unsubscribing Hello netters, just a couple of weeks ago, instructions for unsubscribing to the Histonet were posted. Since I have no intentions of unsubscribing to the Histonet, it was suggested that since I'll be in Toronto for a week, to unsubscribe temporarily. Any one have instruction? Okay, Okay flame away. 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 JNocito <@t> Pathreflab.com Tue Sep 14 15:28:37 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] unsubscribing In-Reply-To: Message-ID: hey thanks, now, all I have to do now is remember my password. Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joe Nocito Sent: Tuesday, September 14, 2004 2:20 PM To: Histonet Subject: [Histonet] unsubscribing Hello netters, just a couple of weeks ago, instructions for unsubscribing to the Histonet were posted. Since I have no intentions of unsubscribing to the Histonet, it was suggested that since I'll be in Toronto for a week, to unsubscribe temporarily. Any one have instruction? Okay, Okay flame away. 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 mcauliff <@t> umdnj.edu Tue Sep 14 19:14:47 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Perferred fixative for animals In-Reply-To: <41470036.30954.8F79EDC3@localhost> References: <41470036.30954.8F79EDC3@localhost> Message-ID: <41478977.9010106@umdnj.edu> If you are not getting good immuno results with formalin fixatives you could try: 1. heat-induced antigen retrieval of your formalin fixed tissues. 2. Fixing in formalin for 4 hours instead of a longer time. 3. Fixing in a fixative that does not cross-link proteins (formalin is a cross-linker) such as Carnoy, methacarn or alcohol with some acetic acid added. I would try the above first. Bear in mind that different antigens have different fixative requirements for good immunoreactivity. Geoff wasielewski.reinhard.von@mh-hannover.de wrote: >Hi Histonetters, >We are looking for the most convenient and morpholgical satisfying fixative for >mouse and rat tissue beside formaldehyde. We want to stain a wide range of >different immunomarkers but don' t like formalin (therefore) too much. Frozen >sections are too bad in morphology. >Please send me any suggestions ! > >Many thanks in advance, >Reinhard. > > > >PD Dr. med. Reinhard von Wasielewski > > >_______________________________________________ >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 S2118495 <@t> student.rmit.edu.au Tue Sep 14 19:15:51 2004 From: S2118495 <@t> student.rmit.edu.au (Christopher Ian Hill) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Embedding of small artery Message-ID: <1095207351.aa8ebbe0S2118495@student.rmit.edu.au> In referance to this, I am an Honours studentat RMIT in Melbourne Aust. My project is looking at atherosclerotic changes in the aorta of mice, so processing and embedding small arteries has become part of my daily routine. My supervisor has worked with the mesenteric vessels of rats, which are smaller again. We use fixation in 10%NBF for 2 hours and then into a processor (alcohol/xyleen) on 15 minute cycles. And then embed the tissue in parafin to give transverse sections of the aorta. Sections are cut at 4um. The tissue sections i use are approx 3-5mm long and need fine tip forceps. I beliece that my supervisor perfused with fixative when she was doing the smaller vessels. Chris Hill BSc Grad. Dip. MLS Quote: hello galina, Rat and mouse organs can be cut easily after short processing(using acetone and xylene)and then embedding in paraffin wax of low melting point(56 C).... same could applied for the small artery also Chris Hill From JQB7 <@t> CDC.GOV Tue Sep 14 18:21:11 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] unsubscribing Message-ID: If I could help you there I'd be in big trouble! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe Nocito Sent: Tue 9/14/2004 4:28 PM To: Joe Nocito; Histonet Cc: Subject: RE: [Histonet] unsubscribing hey thanks, now, all I have to do now is remember my password. Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joe Nocito Sent: Tuesday, September 14, 2004 2:20 PM To: Histonet Subject: [Histonet] unsubscribing Hello netters, just a couple of weeks ago, instructions for unsubscribing to the Histonet were posted. Since I have no intentions of unsubscribing to the Histonet, it was suggested that since I'll be in Toronto for a week, to unsubscribe temporarily. Any one have instruction? Okay, Okay flame away. 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 jnocito <@t> satx.rr.com Tue Sep 14 20:56:12 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Flame me Message-ID: <008e01c49ac7$2db284a0$4661ce44@yourxhtr8hvc4p> Okay, for those of you who are interested or care, on Sunday, I will be wearing a navy blue T-shirt with a red and white target and a star in the center. I'll be the guy with the beard and wearing a flapjacket. Hope everyone has a safe trip to Toronto, watch out for Ivan and LET THE FLAMING BEGIN. See ya Sunday Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX From SJones <@t> cvm.tamu.edu Tue Sep 14 22:23:50 2004 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Flame me Message-ID: Is that the Texas state T-shirt for the competition as well? my bad Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Joe Nocito" 9/14/2004 8:56:12 PM >>> Okay, for those of you who are interested or care, on Sunday, I will be wearing a navy blue T-shirt with a red and white target and a star in the center. I'll be the guy with the beard and wearing a flapjacket. Hope everyone has a safe trip to Toronto, watch out for Ivan and LET THE FLAMING BEGIN. See ya Sunday Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Wed Sep 15 06:12:18 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Flame me References: Message-ID: <002401c49b14$dd8a8330$4661ce44@yourxhtr8hvc4p> no, but now that you mention it, it could be Joe ----- Original Message ----- From: "Sarah Jones" To: ; Sent: Tuesday, September 14, 2004 10:23 PM Subject: Re: [Histonet] Flame me > Is that the Texas state T-shirt for the competition as well? my bad > > > Sarah Jones, HT(ASCP) > Histology Lab > Dept. of Veterinary Integrative Biosciences > College of Veterinary Medicine > Texas A&M University > College Station, TX 77843-4458 > phone: 979-845-3177 > fax: 979-458-3499 > > > >>> "Joe Nocito" 9/14/2004 8:56:12 PM >>> > Okay, for those of you who are interested or care, on Sunday, I will be > wearing a navy blue T-shirt with a red and white target and a star in > the center. I'll be the guy with the beard and wearing a flapjacket. > Hope everyone has a safe trip to Toronto, watch out for Ivan and LET THE > FLAMING BEGIN. > > See ya Sunday > > > 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 kmerriam2003 <@t> yahoo.com Wed Sep 15 06:51:40 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] offtopic: histologist on "the Benefactor" Message-ID: <20040915115140.51970.qmail@web52508.mail.yahoo.com> Hello all, I know most of you are getting ready for NSH. But did anyone watch "the Benefactor"? One of the contestants is a Histologist from Portland, OR! How cool is that! Kim Merriam Novartis Cambridge, MA Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! From tobetrice <@t> yahoo.com Wed Sep 15 07:55:16 2004 From: tobetrice <@t> yahoo.com (Patrice McCall) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] need help with pan-keratin stain (HTL) Message-ID: <20040915125516.24822.qmail@web40006.mail.yahoo.com> hey, does anyone know how to do a pan-keratin stain manually? i am taking the HTL, but i work in a research histology lab and we do not have immunohistochemistry equipment. i need to know all the materials to buy and the procedures for doing the stain. thanks to anyone that can help __________________________________ Do you Yahoo!? New and Improved Yahoo! Mail - Send 10MB messages! http://promotions.yahoo.com/new_mail From la.sebree <@t> hosp.wisc.edu Wed Sep 15 08:17:24 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] need help with pan-keratin stain (HTL) Message-ID: The short answer Patrice is to buy a manual staining kit and the antibody. Try Dako for both. 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: Patrice McCall [mailto:tobetrice@yahoo.com] Sent: Wednesday, September 15, 2004 7:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] need help with pan-keratin stain (HTL) hey, does anyone know how to do a pan-keratin stain manually? i am taking the HTL, but i work in a research histology lab and we do not have immunohistochemistry equipment. i need to know all the materials to buy and the procedures for doing the stain. thanks to anyone that can help __________________________________ Do you Yahoo!? New and Improved Yahoo! Mail - Send 10MB messages! http://promotions.yahoo.com/new_mail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From plucas <@t> biopath.org Wed Sep 15 08:30:22 2004 From: plucas <@t> biopath.org (Paula Lucas) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Sulfur Granule Stain Message-ID: <01C49AED.7B258680.plucas@biopath.org> The pathologist is requesting a sulfur granule stain on a peritoneal fluid. We prepared a thinprep slide for this. Could you tell me the best stain for this? Thank you, Paula Lucas Bio-Path Medical Group From juan.gutierrez <@t> christushealth.org Wed Sep 15 08:46:12 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] RE: Histonet Digest, Vol 10, Issue 19 Message-ID: Your Ventana rep should be able and willing to set up your validation protocols, and help you get started. Good luck. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 -----Original Message----- From: Shawn Salesky [mailto:ssalesky@LowellGeneral.org] Sent: Tuesday, September 14, 2004 12:39 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: Histonet Digest, Vol 10, Issue 19 Hello All, The Lab I work in is currently setting up automated IHC, ISH and Special stains on Ventana systems. First question... Is anyone using Ventana Inform HPV HR in a clinical setting? and How did you validate? Second question... Is there a standard number of cases to run in validations of Immuno's? Third question... What numbers did you use to validate your special stains? I'm pretty much at a loss, as I cannot find any literature on the subjects. Thanks in advance, Shawn Salesky TC LGH Histo > ---------- > From: > histonet-bounces@lists.utsouthwestern.edu[SMTP:histonet-bounces@lists.utso > uthwestern.edu] on behalf of > histonet-request@lists.utsouthwestern.edu[SMTP:histonet-request@lists.utso > uthwestern.edu] > Sent: Tuesday, September 14, 2004 1:08 PM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet Digest, Vol 10, Issue 19 > > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. Re: Anti-parrafin floor (DDittus787@aol.com) > 2. re embedding of small artery (carl hobbs) > 3. Recover tissue sections on non-plus slides (Kim Byrnes) > 4. RE: Recover tissue sections on non-plus slides (Bonnie Whitaker) > 5. Re: Recover tissue sections on non-plus slides > (chiggerson@memhosp.com) > 6. alizarin red fading (Julien) > 7. fluorescent dyes/lables for neuronal activities? (Cao, Xudong) > 8. RE: alizarin red fading (Barry R Rittman) > 9. RE: Recover tissue sections on non-plus slides (Laurie Colbert) > 10. cryomacrocut (Robin Turcotte) > 11. cryomacrocut (Robin Turcotte) > 12. cryomacrocut (Robin Turcotte) > 13. cryomacrocut (Robin Turcotte) > 14. cryomacrocut (Robin Turcotte) > 15. Shandon Hypercenter Xp (Jim Staruk) > 16. Superfrost Slides (Bill Sinai) > 17. embedding of small artery_paraffin (Dr. Raghul) > 18. vascular graft_sectioning difficulty (Dr. Raghul) > 19. Test (Dimaano, Nena) > 20. RE: Superfrost Slides (Joe Nocito) > 21. Perferred fixative for animals > (wasielewski.reinhard.von@mh-hannover.de) > 22. RE: Fixative for animals (Cullen, Kay) > 23. Histo. Assistant Job Descriptions (Amy Self) > 24. Re: Perferred fixative for animals (LaCinda Burchell) > 25. Re: Recover tissue sections on non-plus slides (Barbara Lentz) > 26. RE: Histo. Assistant Job Descriptions (Kari Bradshaw) > 27. FW: [Histonet] Histo. Assistant Job Descriptions (Bonnie Whitaker) > 28. rat embryos (Jeffus, Brandon) > 29. RE: FW: [Histonet] Histo. Assistant Job Descriptions > (Bonnie Whitaker) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Mon, 13 Sep 2004 13:48:40 -0400 > From: DDittus787@aol.com > Subject: [Histonet] Re: Anti-parrafin floor > To: BBEIER@kumc.edu ("Barbara Beier") > Cc: histonet@pathology.swmed.edu > Message-ID: <7B0A3F7A.56A40CDC.0A1F969F@aol.com> > Content-Type: text/plain; charset=iso-8859-1 > > Barb: > the flooring came from Acme Panels-the company is in the UK and can be > found on Google, it is anti-solvent, chemical, paraffin,etc. It comes in 3 > or 4 colors and everyone loves it. > We had a contractor bring it in and install it. > dana > > > > ------------------------------ > > Message: 2 > Date: Mon, 13 Sep 2004 19:02:14 +0100 > From: "carl hobbs" > Subject: [Histonet] re embedding of small artery > To: "Histonet" > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > In my experienc, with fixed tissues it is easy to embed using agarose, > then > to re-orient the tissue for frozen/pwax sectioning: Dissolve 1% agarose( > aq) > and when about 50C place the tissue in the agarose, in a bigger mould. It > will set quickly.....better if placed at 4C for a while. Remove the set > agarose and cut away excess agarose so you have the tissue in the > orientation of your choice. Then process to pwax. Or, place in OCT for a > couple of hours on a rotator. Then embed in an OCT filled mould in the > orientation you need and....freeze. If you are doing fresh tissue, get the > melted, cooling agarose into a mould , place the tissue and cool quickly. > Immediately place in OCT, swirl gently for a minute, then freeze. Play > around with control material until you are confident of the technique and > to > reassure yourself that the elevated temp does not adversely afffect your > tissue. You can also do gelatin/vibratome sections of the fixed tissue, if > you have a vibratome. > --- > Outgoing mail is certified Virus Free. > Checked by AVG anti-virus system (http://www.grisoft.com). > Version: 6.0.754 / Virus Database: 504 - Release Date: 06/09/2004 > > > > > ------------------------------ > > Message: 3 > Date: Mon, 13 Sep 2004 11:34:29 -0700 (PDT) > From: Kim Byrnes > Subject: [Histonet] Recover tissue sections on non-plus slides > To: histonet@lists.utsouthwestern.edu > Message-ID: <20040913183429.84652.qmail@web12308.mail.yahoo.com> > Content-Type: text/plain; charset=us-ascii > > Does anyone have a method or know of a product that > can recover or rescue tissue sections that were > accidentally mounted onto non-plus (non-polarized, > non-coated) slides? We would like to do immuno on > these slides, but they consistently fall off of the > slides. > > Any ideas? > > Thanks, > > Kim > Georgetown University > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - 50x more storage than other providers! > http://promotions.yahoo.com/new_mail > > > > ------------------------------ > > Message: 4 > Date: Mon, 13 Sep 2004 14:24:14 -0500 > From: "Bonnie Whitaker" > Subject: RE: [Histonet] Recover tissue sections on non-plus slides > To: "'Kim Byrnes'" > Cc: histonet@lists.utsouthwestern.edu > Message-ID: <000301c499c7$41758920$3601a8c0@brownpathology.net> > Content-Type: text/plain; charset="us-ascii" > > There is a procedure to remove tissue sections from a slide, and they can > then be mounted on + slides. The product that works best in my hands is > Mount-Quick. It has more than one distributer, but I know that Newcomer > Supply (800-383-7799) has this product, as well as directions on how to do > the procedure. > > Bonnie Whitaker > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Byrnes > Sent: Monday, September 13, 2004 1:34 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Recover tissue sections on non-plus slides > > Does anyone have a method or know of a product that > can recover or rescue tissue sections that were > accidentally mounted onto non-plus (non-polarized, > non-coated) slides? We would like to do immuno on > these slides, but they consistently fall off of the > slides. > > Any ideas? > > Thanks, > > Kim > Georgetown University > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - 50x more storage than other providers! > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 5 > Date: Mon, 13 Sep 2004 14:46:54 -0500 > From: chiggerson@memhosp.com > Subject: Re: [Histonet] Recover tissue sections on non-plus slides > To: histonet@lists.utsouthwestern.edu > Cc: Kim Byrnes > Message-ID: > > Content-Type: text/plain; charset="US-ASCII" > > Kim, > > The 2000 Tech Sample exercise HT-1 gives a procedure for this. The title > of the exercise is "Use Mount Quick to Remove Tissue Sections from > Non-Positively Charged Slides for Immunohistochemical and Special Stains". > > > Good luck! > > Cindy > > Cindy Higgerson HTL(ASCP) > Surgical Pathology Supervisor > Memorial Hospital > > > > > > Kim Byrnes > Sent by: histonet-bounces@lists.utsouthwestern.edu > 09/13/2004 01:34 PM > > To > histonet@lists.utsouthwestern.edu > cc > > Subject > [Histonet] Recover tissue sections on non-plus slides > > > > > > > Does anyone have a method or know of a product that > can recover or rescue tissue sections that were > accidentally mounted onto non-plus (non-polarized, > non-coated) slides? We would like to do immuno on > these slides, but they consistently fall off of the > slides. > > Any ideas? > > Thanks, > > Kim > Georgetown University > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - 50x more storage than other providers! > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 6 > Date: Mon, 13 Sep 2004 16:58:59 -0400 > From: "Julien" > Subject: [Histonet] alizarin red fading > To: > Message-ID: <00c501c499d4$7dc51140$9310d784@uqar.qc.ca> > Content-Type: text/plain; charset="iso-8859-1" > > Hello all, > > We do differencial staining of bone and cartillage on small fish (~1cm). > To do so, we use the Clear and Stain technique which results in red bone > coloration and blue cartillage coloration. After a month or so, We noticed > that the red stain tends to fade off. We tried to re-stain the fish, but > the alizarin red wouldn't stain the bones anymore. > > Why is this so? Is it that bone decalcifies in the glycerin? We put a few > cristals of thymol in the glycerin to avoid mold development. Could this > be a decalcifying agent? Does anybody know of a way to re-stain? > > For the time being, we score bony elements within 3 days of staining to > avoid fading problems, but this stops us from doing batch staining. Batch > staining would be much faster. We would then keep stained fish in glycerin > until scoring. > > Thanks for any suggestion. > > Julien Lambrey de Souza > Biologie ?volutive, > Universit? du Qu?bec ? Rimouski, > > > Julien Lambrey de Souza > Biologie ?volutive, > Universit? du Qu?bec ? Rimouski, > > (418) 723-1986 #1438 > > > ------------------------------ > > Message: 7 > Date: Mon, 13 Sep 2004 17:03:50 -0400 > From: "Cao, Xudong" > Subject: [Histonet] fluorescent dyes/lables for neuronal activities? > To: > Message-ID: > <1DA88DDC48CCC245A777599FDAEED6D0A3909B@MAIL1.AD.Brown.Edu> > Content-Type: text/plain; charset="iso-8859-1" > > Greetings! first of all I'd like to thank eveybody who responded to my > previous posts. The advice/ideas I received were valuable! Here I have > one more question: I am studying nerve innervation into artificial skin > graft that we make in vitro and my immunostaining tells me that there are > some wonderful nerve fibers growing into the skin. The question that we > have now is if the innervation functional. In order to test this, we want > to mechanically tease the artificial skin and see if there is some > responses from the nerve cells that have presumablly innervated the > corresponding part of the skin. Thus, it would be nice to find some > fluorescent dyes/lables that will glow when the neurons are > activated/firing. any suggestions? > > best regards, > > Xudong > > > > ------------------------------ > > Message: 8 > Date: Mon, 13 Sep 2004 16:29:32 -0500 > From: "Barry R Rittman" > Subject: RE: [Histonet] alizarin red fading > To: "histonet" > Message-ID: > <566FB0B522443D43AF02D2ADBE35A6F0F13357@UTHEVS3.mail.uthouston.edu> > Content-Type: text/plain; charset="iso-8859-1" > > I cannot see why the addition of a few crystals of thymol would make the > glycerin acidic enough to cause removal of calcium and thus leaching of > the alizarin. > If you are using 100% glycerin, there should be no need for the addition > of thymol. > I am not sure exactly how you have processed these specimens. If you are > using the classical method with potassium hydroxide (as the leaching agent > to remove excess stain from soft tissues), is it possible that there were > traces of potassium hydroxide left in the specimen? > I have some specimens that were stained in 1961 and some in 1970 that show > no signs of fading. > Barry > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Julien > Sent: Monday, September 13, 2004 3:59 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] alizarin red fading > > Hello all, > > We do differencial staining of bone and cartillage on small fish (~1cm). > To do so, we use the Clear and Stain technique which results in red bone > coloration and blue cartillage coloration. After a month or so, We noticed > that the red stain tends to fade off. We tried to re-stain the fish, but > the alizarin red wouldn't stain the bones anymore. > > Why is this so? Is it that bone decalcifies in the glycerin? We put a few > cristals of thymol in the glycerin to avoid mold development. Could this > be a decalcifying agent? Does anybody know of a way to re-stain? > > For the time being, we score bony elements within 3 days of staining to > avoid fading problems, but this stops us from doing batch staining. Batch > staining would be much faster. We would then keep stained fish in glycerin > until scoring. > > Thanks for any suggestion. > > Julien Lambrey de Souza > Biologie ?volutive, > Universit? du Qu?bec ? Rimouski, > > > Julien Lambrey de Souza > Biologie ?volutive, > Universit? du Qu?bec ? Rimouski, > > (418) 723-1986 #1438 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 9 > Date: Mon, 13 Sep 2004 14:43:49 -0700 > From: "Laurie Colbert" > Subject: RE: [Histonet] Recover tissue sections on non-plus slides > To: "Kim Byrnes" , "Histonet (E-mail)" > > Message-ID: > > <0BE6ADFAE4E7E04496BF21ABD346628001C5C062@EXCHANGE1.huntingtonhospital.com > > > > Content-Type: text/plain; charset="iso-8859-1" > > We have used the Mount Quick from Newcomer Supply, and it works well. > > Laurie Colbert > Huntington Memorial Hospital > Pasadena, CA > > -----Original Message----- > From: Kim Byrnes [mailto:k_byrnes@yahoo.com] > Sent: Monday, September 13, 2004 11:34 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Recover tissue sections on non-plus slides > > > Does anyone have a method or know of a product that > can recover or rescue tissue sections that were > accidentally mounted onto non-plus (non-polarized, > non-coated) slides? We would like to do immuno on > these slides, but they consistently fall off of the > slides. > > Any ideas? > > Thanks, > > Kim > Georgetown University > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - 50x more storage than other providers! > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 10 > Date: Mon, 13 Sep 2004 20:01:58 -0400 > From: "Robin Turcotte" > Subject: [Histonet] cryomacrocut > To: > Message-ID: <000701c499ee$cb688f70$df8ceccf@client> > Content-Type: text/plain; charset="iso-8859-1" > > Does anyone have an address from a compagny or an hospital who have a used > cryomacrocut in good condition to be sold. > > Thank you > > Robin > > > > > ------------------------------ > > Message: 11 > Date: Mon, 13 Sep 2004 20:07:19 -0400 > From: "Robin Turcotte" > Subject: [Histonet] cryomacrocut > To: > Message-ID: <000e01c499ee$ee9cdbe0$df8ceccf@client> > Content-Type: text/plain; charset="iso-8859-1" > > Does anyone have an address from a compagny or an hospital who have a used > cryomacrocut in good condition to be sold. > > Thank you > > Robin > > > > > ------------------------------ > > Message: 12 > Date: Mon, 13 Sep 2004 20:07:47 -0400 > From: "Robin Turcotte" > Subject: [Histonet] cryomacrocut > To: > Message-ID: <000f01c499ee$ef064c10$df8ceccf@client> > Content-Type: text/plain; charset="iso-8859-1" > > Does anyone have an address from a compagny or an hospital who have a used > cryomacrocut in good condition to be sold. > > Thank you > > Robin > > > > > ------------------------------ > > Message: 13 > Date: Mon, 13 Sep 2004 20:10:32 -0400 > From: "Robin Turcotte" > Subject: [Histonet] cryomacrocut > To: > Message-ID: <001301c499ef$45100dd0$df8ceccf@client> > Content-Type: text/plain; charset="iso-8859-1" > > Does anyone have an address from a compagny or an hospital who have a used > cryomacrocut in good condition to be sold. > > Thank you > > Robin > > > > > ------------------------------ > > Message: 14 > Date: Mon, 13 Sep 2004 20:11:36 -0400 > From: "Robin Turcotte" > Subject: [Histonet] cryomacrocut > To: > Message-ID: <001c01c499ef$6fce96e0$df8ceccf@client> > Content-Type: text/plain; charset="iso-8859-1" > > Does anyone have an address from a compagny or an hospital who have a used > cryomacrocut in good condition to be sold. > > Thank you > > Robin > > > > ------------------------------ > > Message: 15 > Date: Mon, 13 Sep 2004 21:15:52 -0400 > From: "Jim Staruk" > Subject: [Histonet] Shandon Hypercenter Xp > To: > Message-ID: <000201c499f8$87d3cb80$6401a8c0@yourw04gtxld67> > Content-Type: text/plain; charset="us-ascii" > > I'm looking for someone familiar with the Shandon Hypercenter Xp tissue > processor. I need some technical advise. Can someone help me? Anyone > planning to invoice me for their help need not reply. > > Thank you > > Jim > > ________________________ > James E. Staruk, HT(ASCP) > Mass Histopathology Service > www.masshistology.com > > > > > > ------------------------------ > > Message: 16 > Date: Tue, 14 Sep 2004 13:43:17 +1000 > From: "Bill Sinai" > Subject: [Histonet] Superfrost Slides > To: "histonet \(E-mail\)" > Message-ID: <000101c49a0c$f9356e50$e1ce080a@wsahs.nsw.gov.au> > Content-Type: text/plain; charset="iso-8859-1" > > > Is anyone having troubles with NO staining on some slides on the Ventana > Benchmark. > The biggest problem seems to be with the EDTA buffered CC1 retrieval > solution. > Not all slides are effected, just an occasional one, but we usually end up > with little or no staining with anything including the Haematoxylin. > We use Menzel Superforst Plus and Superfrost Plus Ultra, and both slides > have shown the effect. > > Thanks > Bill Sinai > Laboratory Manager > Tissue Pathology, ICPMR > Westmead NSW 2145 > Australia > Ph 02 9845 7774 > > > __________________________________________________________________ > > This electronic message and any attachments may be confidential. If you > are not the intended recipient of this message would you please delete the > message and any attachments and advise the sender. 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. > > > > ------------------------------ > > Message: 17 > Date: Tue, 14 Sep 2004 16:31:15 +0530 > From: "Dr. Raghul" > Subject: [Histonet] embedding of small artery_paraffin > To: histonet@lists.utsouthwestern.edu > Message-ID: > > hello galina, > > Rat and mouse organs can be cut easily after short processing(using > acetone and xylene)and then embedding in paraffin wax of low melting point > (56 C). We have followed the protocols from below given reference > successfully and same could applied for the small artery also > > Theory and practice of Histological techniques (bancroft and steven) > > (or) > > Manual of histologic staining methods of the armed forces institutes of > pathology edited by Lee G. Luna, III edition page 16 > > > Raghul J > Histopathology-Implant Biology > Sree chitra tirunal institute > Thiruvananthapuram 695 012 > > > > > ------------------------------ > > Message: 18 > Date: Tue, 14 Sep 2004 16:56:23 +0530 > From: "Dr. Raghul" > Subject: [Histonet] vascular graft_sectioning difficulty > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > hello members, > > We cut 5 ?m thin sections of paraffin embedded vascular graft(Dacron) in > a rotary microtome with disposable blades but the senior tech tells that > the tissue sections are crumbling and forces us to go for thick sections > around 10?m but again not with much success. > > We can understand that there is not enough tissue adhereing to the > graft and we are literally cutting the graft. But did anyone had such > problems sectioning vascular grafts. Any suggestions? > > > regards , > > Raghul J > scientist > Histopathology- implant biology division > Srichitra tirunal institute of medical sciences and technology > Trivandrum -695 012 > Kerala, India > > > > > ------------------------------ > > Message: 19 > Date: Tue, 14 Sep 2004 07:54:55 -0400 > From: "Dimaano, Nena" > Subject: [Histonet] Test > To: > Message-ID: > > <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD06210EE4@HOS2KEXCHCL.howost.strykercorp.c > om> > > Content-Type: text/plain; charset="iso-8859-1" > > > Nena Dimaano > Advanced Technology > Stryker Orthopaedics > 325 Corporate Drive > Mahwah, NJ 07430 > tel: 201-831-5338 > fax: 201-831-6224 > email: Nena.Dimaano@stryker.com > > > > ------------------------------ > > Message: 20 > Date: Tue, 14 Sep 2004 07:09:09 -0500 > From: "Joe Nocito" > Subject: RE: [Histonet] Superfrost Slides > To: "Bill Sinai" , "histonet > \(E-mail\)" > > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > Bill, > we've had problems also. It could be that the slides might be too changed, > which is causing the reagents to bead up. Currently, we are using positive > charged slides from Statlab Medical Products, which I think they get from > Erie Scientific. Statlab's number is 1-800-442-3573. The may be able to > direct you to some one closer to you. > > 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 Bill > Sinai > Sent: Monday, September 13, 2004 10:43 PM > To: histonet (E-mail) > Subject: [Histonet] Superfrost Slides > > > > Is anyone having troubles with NO staining on some slides on the Ventana > Benchmark. > The biggest problem seems to be with the EDTA buffered CC1 retrieval > solution. > Not all slides are effected, just an occasional one, but we usually end up > with little or no staining with anything including the Haematoxylin. > We use Menzel Superforst Plus and Superfrost Plus Ultra, and both slides > have shown the effect. > > Thanks > Bill Sinai > Laboratory Manager > Tissue Pathology, ICPMR > Westmead NSW 2145 > Australia > Ph 02 9845 7774 > > > __________________________________________________________________ > > This electronic message and any attachments may be confidential. If you > are not the intended recipient of this message would you please delete the > message and any attachments and advise the sender. 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. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 21 > Date: Tue, 14 Sep 2004 14:29:10 +0200 > From: wasielewski.reinhard.von@mh-hannover.de > Subject: [Histonet] Perferred fixative for animals > To: histonet@lists.utsouthwestern.edu > Message-ID: <41470036.30954.8F79EDC3@localhost> > Content-Type: text/plain; charset=US-ASCII > > Hi Histonetters, > We are looking for the most convenient and morpholgical satisfying > fixative for > mouse and rat tissue beside formaldehyde. We want to stain a wide range of > > different immunomarkers but don' t like formalin (therefore) too much. > Frozen > sections are too bad in morphology. > Please send me any suggestions ! > > Many thanks in advance, > Reinhard. > > > > PD Dr. med. Reinhard von Wasielewski > > > > > ------------------------------ > > Message: 22 > Date: Tue, 14 Sep 2004 08:56:22 -0400 > From: "Cullen, Kay" > Subject: [Histonet] RE: Fixative for animals > To: "Histonet \(E-mail\)" > Message-ID: > > > Content-Type: text/plain; charset="iso-8859-1" > > Are you busing straight formaldehyde or formaldehyde/glutaraldehyde? I > get very good results with Bouin's for most tissue. > > Kay Cullen > Research Associate > Department of Cell Biology > University of Massachusetts Medical School > Worcester, MA USA > > > > ------------------------------ > > Message: 23 > Date: Tue, 14 Sep 2004 10:05:39 -0400 > From: "Amy Self" > Subject: [Histonet] Histo. Assistant Job Descriptions > To: > Message-ID: > <39836CD6DB61654E8F95A35898C921860A87F3@exchange.gmhpost.com> > Content-Type: text/plain; charset="iso-8859-1" > > > > Good Morning Netters, > > We are looking to hire a histology assistant in our lab. I was > wandering if anyone had any job descriptions as well as job requirements > that they could share with me? We have one but it very minimal as to what > the histo assistant should and should not do so our lab manager would like > to see what other hospitals are doing and requiring for this job position. > Thanks in advance, amy > > > > > > > NOTE: > The information contained in this message may be privileged, 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 this message and deleting it from > your computer. Thank you. > > > ------------------------------ > > Message: 24 > Date: Tue, 14 Sep 2004 09:16:54 -0500 > From: "LaCinda Burchell" > Subject: Re: [Histonet] Perferred fixative for animals > To: , > > Message-ID: > Content-Type: text/plain; charset=US-ASCII > > Dear Reinhard, > We use Prefer fixative from Anatech Ltd, www.anatechltdusa. We have > beautiful morphology, staining is wonderful (specials and immunos). The > only down side of Prefer is that it affects the staining of eosinophils. > We've recently learned how to get around that issue in human tissues, > but haven't figured it out in rat tissues. Best of luck! Cindy B > > LaCinda Burchell, BA, AS, HT(ASCP) > University of Wisconsin-Madison, Medical School > Asthma and Allergy Research IHC Lab > 600 Highland Ave. CSC K4/913 > Madison, Wisconsin 53792 > > Phone: 608-262-3518 > FAX: 608-263-3746 > > >>> 09/14/04 07:29AM >>> > Hi Histonetters, > We are looking for the most convenient and morpholgical satisfying > fixative for > mouse and rat tissue beside formaldehyde. We want to stain a wide range > of > different immunomarkers but don' t like formalin (therefore) too much. > Frozen > sections are too bad in morphology. > Please send me any suggestions ! > > Many thanks in advance, > Reinhard. > > > > PD Dr. med. Reinhard von Wasielewski > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 25 > Date: Tue, 14 Sep 2004 10:48:20 -0400 > From: "Barbara Lentz" > Subject: Re: [Histonet] Recover tissue sections on non-plus slides > To: , > Message-ID: > Content-Type: text/plain; charset=US-ASCII > > We, too, use the Mount Quick from Newcomer Supply. On a regular basis, > we remove H&E tissue sections to perform immunos. > www.newcomersupply.com is their address, Barb > > >>> Kim Byrnes 09/13/04 02:34PM >>> > Does anyone have a method or know of a product that > can recover or rescue tissue sections that were > accidentally mounted onto non-plus (non-polarized, > non-coated) slides? We would like to do immuno on > these slides, but they consistently fall off of the > slides. > > Any ideas? > > Thanks, > > Kim > Georgetown University > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - 50x more storage than other providers! > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 26 > Date: Tue, 14 Sep 2004 07:41:17 -0700 > From: "Kari Bradshaw" > Subject: RE: [Histonet] Histo. Assistant Job Descriptions > To: "Amy Self" , > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > Here is our job description. > > :) Kari Bradshaw, HT(ASCP) > Laboratory Manager > Lower Columbia Pathologists > 1217 14th Ave > Longview, WA 98632 > (360)425-5620 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amy Self > Sent: Tuesday, September 14, 2004 7:06 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histo. Assistant Job Descriptions > > > > > Good Morning Netters, > > We are looking to hire a histology assistant in our lab. I was > wandering > if anyone had any job descriptions as well as job requirements that they > could share with me? We have one but it very minimal as to what the histo > assistant should and should not do so our lab manager would like to see > what > other hospitals are doing and requiring for this job position. Thanks in > advance, amy > > > > > > > NOTE: > The information contained in this message may be privileged, 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 this message and deleting it from your > computer. Thank you. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ > > Message: 27 > Date: Tue, 14 Sep 2004 10:54:03 -0500 > From: "Bonnie Whitaker" > Subject: FW: [Histonet] Histo. Assistant Job Descriptions > To: > Message-ID: <000801c49a73$0ee10140$3601a8c0@brownpathology.net> > Content-Type: text/plain; charset="us-ascii" > > Here is ours. > Bonnie Whitaker > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kari > Bradshaw > Sent: Tuesday, September 14, 2004 9:41 AM > To: Amy Self; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histo. Assistant Job Descriptions > > Here is our job description. > > :) Kari Bradshaw, HT(ASCP) > Laboratory Manager > Lower Columbia Pathologists > 1217 14th Ave > Longview, WA 98632 > (360)425-5620 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amy Self > Sent: Tuesday, September 14, 2004 7:06 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histo. Assistant Job Descriptions > > > > > Good Morning Netters, > > We are looking to hire a histology assistant in our lab. I was > wandering > if anyone had any job descriptions as well as job requirements that they > could share with me? We have one but it very minimal as to what the histo > assistant should and should not do so our lab manager would like to see > what > other hospitals are doing and requiring for this job position. Thanks in > advance, amy > > > > > > > NOTE: > The information contained in this message may be privileged, 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 this message and deleting it from your > computer. Thank you. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ > > Message: 28 > Date: Tue, 14 Sep 2004 11:33:48 -0500 > From: "Jeffus, Brandon" > Subject: [Histonet] rat embryos > To: > Message-ID: > <748D4F173E342D47B69FC23C87E5913C0125E7D4@EXCHANGE4.ad.uams.edu> > Content-Type: text/plain; charset="us-ascii" > > Hello all... im rather new to immunohistochemistry so bear with me. I'm > attempting to process some rat embryos for staining with a primary Ab > and then a Cy3 labeled secondary for visualization of protein > localization. I'm wondering if the best processing method would be > frozen sections of the samples or paraffin embedding the samples? Any > information would be greatly appreciated. > > > > Brandon C Jeffus > > Graduate Student > > > > ------------------------------ > > Message: 29 > Date: Tue, 14 Sep 2004 11:40:24 -0500 > From: "Bonnie Whitaker" > Subject: RE: FW: [Histonet] Histo. Assistant Job Descriptions > To: "'Nita Searcy'" > Cc: histonet@lists.utsouthwestern.edu > Message-ID: <000001c49a79$88e006c0$3601a8c0@brownpathology.net> > Content-Type: text/plain; charset="us-ascii" > > > OOPS... attachments bad!! Pasting good. > > > Below is our aid job description. > > Bonnie Whitaker > > > Job Description for Laboratory/Grossing Aid > > > > > Reports To: Laboratory Manager > > Pathologists > > > > Qualifications: > > > > Education: high school student or have > diploma > or GED > > > > Experience: not required > > > > Certification: not required > > > > Personal: The Lab/Grossing Aid must > demonstrate an ability to listen carefully and follow directions well. > The > incumbent must quickly grasp the work-flow of specimen accessioning and > grossing and become familiar with these procedures. The incumbent must > demonstrate reasonable intelligence, diligence, accuracy, cooperation, > neatness, and promptness in keeping with the prestige and image of this > laboratory. The incumbent takes guidance and instruction from the > pathologists, with whom they are working, as well as the Laboratory > Manager > and Executive Director. > > Responsibilities: The Lab/Grossing Aid will be > responsible for the following duties: > > 1. Accessioning specimens and assisting with grossing procedures > including: cassette labeling, pulling specimens for additional tissue > submission, and discarding specimens in accordance with policy/procedure > manual. > > 2. Assisting with the performance of routine maintenance and QC > procedures on equipment to include but not limited to cryostat, grossing > area, stain setup and filters. > > 3. Maintenance of inventory > > 4. Routine changing of stains with appropriate records maintenance. > > 5. Cleaning of the grossing area and instruments. > > 6. Any filing or routine clerical duties necessary. > > 7. Other duties as assigned. > > > > > General: > > 1. The Lab/Grossing Aid shall be responsible for maintaining all > company > records of a proprietary or confidential nature secure and confidential > and > shall return all lists of clients, potential clients, and all other > records, > at Brown & Associates request. > > 2. The Lab/Grossing Aid is responsible for representing B&AML in a > professional and dignified manner befitting the status of the laboratory > in > the Houston Medical Community. > > 3. The Lab/Grossing Aid will be expected to comply with all policies > of > B&AML as stated in the employee handbook and any newly instituted policies > not specifically stated in the handbook. Will be expected to keep > assigned > work area as neat and clean as possible so as to afford the maximum > functional usage of the assigned area. Will be expected to keep all > records > and files available and accessible to authorized personnel at all times. > > 4. Lab/Grossing Aid must be in reasonable good health with no record > of > chronic illness resulting in undue absence from work. > > > > I, the undersigned, have carefully read the above job description and > fully > understand all that is stated herein. I agree to perform all of the > duties > above as well as any other duties deemed necessary by my supervisor within > the limits described here. > > > > > > _________________________________________ > _____________________ > > > EMPLOYEE SIGNATURE > DATE > > > > > > > > _________________________________________ > _____________________ > > LABORATORY MANAGER DATE > > > > > > > > > > Bonnie Whitaker > > -----Original Message----- > From: Nita Searcy [mailto:NSEARCY@swmail.sw.org] > Sent: Tuesday, September 14, 2004 11:30 AM > To: bwhitaker@brownpathology.com > Subject: Re: FW: [Histonet] Histo. Assistant Job Descriptions > > > > Bonnie, I can't view your description- could you fax? 254-724-4391 > > Thanks! > > >>> "Bonnie Whitaker" 9/14/2004 10:54:03 AM > >>> > > Here is ours. > Bonnie Whitaker > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Kari > Bradshaw > Sent: Tuesday, September 14, 2004 9:41 AM > To: Amy Self; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histo. Assistant Job Descriptions > > Here is our job description. > > :) Kari Bradshaw, HT(ASCP) > Laboratory Manager > Lower Columbia Pathologists > 1217 14th Ave > Longview, WA 98632 > (360)425-5620 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On > Behalf Of Amy > Self > Sent: Tuesday, September 14, 2004 7:06 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histo. Assistant Job Descriptions > > > > > Good Morning Netters, > > We are looking to hire a histology assistant in our lab. I was > wandering > if anyone had any job descriptions as well as job requirements that they > could share with me? We have one but it very minimal as to what the histo > assistant should and should not do so our lab manager would like to see > what > other hospitals are doing and requiring for this job position. Thanks in > advance, amy > > > > > > > NOTE: > The information contained in this message may be privileged, 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 this 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 > > End of Histonet Digest, Vol 10, Issue 19 > **************************************** > This e-mail and any attachments is intended only for the person or entity to which it is addressed and may contain confidential and privileged information. If you are not the intended recipient, please notify the sender immediately by return e-mail, delete this e-mail and destroy any copies. Any dissemination or use of this information by a person other than the intended recipient is unauthorized and may be illegal. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Charles.Embrey <@t> carle.com Wed Sep 15 08:48:36 2004 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Flame me Message-ID: Hey Joe, I know it has been a while since we retired..... but I remember wearing a flak jacket. Does the flapjacket protect you from flapjacks? Sorry Joe, I just couldn't resist. Have fun in Canada. I was there 3 years ago for the AAPA Conference and I had a blast. Chuck Embrey -----Original Message----- From: Joe Nocito [mailto:jnocito@satx.rr.com] Sent: Tuesday, September 14, 2004 8:56 PM To: histonet Subject: [Histonet] Flame me Okay, for those of you who are interested or care, on Sunday, I will be wearing a navy blue T-shirt with a red and white target and a star in the center. I'll be the guy with the beard and wearing a flapjacket. Hope everyone has a safe trip to Toronto, watch out for Ivan and LET THE FLAMING BEGIN. See ya Sunday 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 vandries <@t> vub.ac.be Wed Sep 15 08:52:07 2004 From: vandries <@t> vub.ac.be (Veronique Andriessen) Date: Fri Sep 16 15:24:02 2005 Subject: [Histonet] Glucose oxidase Message-ID: Hi histonetters, I've got a problem. I'm have to do immunostaining with HRP and AP on inflamed rat pancreas (fresh frozen sections, fixed with acetone ethanol 3:1). I have to quench the endogenous peroxidase in the white blood cells. I have tried the glucose oxidase block as described in the histonet archives, but I am not able to get it working. I have ordered the correct ingredients from Sigma, but nothing! The only thing that is not recently purchased is the Sodium azide, but I can't imagine this going bad. I even tried adding 0.1% saponin, but this also did't help. I really need some advice, I'm getting desparate. Kindest regards, Veronique Andriessen BAS Lab. Molecular Liver Cell Biology Free University Brussels (VUB), Belgium Tel: 0032-2-477.4259 Fax: 0032-2-477.4412 vandries@vub.ac.be GLUCOSE OXIDASE BLOCK FOR ENDOGENOUS PEROXIDASE/PSEUDOPEORXIDASES IN FROZEN/PARAFFIN SECTIONS Reference: Andrew SM, Jasani B. An improved method for the inhibition of endogenous peroxidase non-deleterious to lymphocyte surface markers. Application to immunoperoxidase studies on eosinophil-rich tissue preparations. Histochemical Journal 19:426-30, 1987 All forms of endogenous peroxidase may not be inhibited by the usual methanol/hydrogen peroxide or buffer/hydrogen peroxide blocking mixtures. This method produces nascent hydrogen peroxide that is preferable to the normal methods using preformed hydrogen peroxide (which you add or buy in your endogenous peroxide blocker). This new method to block 'peroxidatic activity' is consistently complete. This method is an enzymatic reaction that produces a slow, steady, very low concentration of hydrogen peroxide. It is particularly useful for monoclonal antibody staining on frozen sections that are minimally fixed with acetone. Procedure: Glucose (Sigma G 5250) 0.180 g Glucose oxidase (Sigma G 6641) 0.005 g Sodium azide 0.0065g DPBS 50 ml This procedure has doubled concentration of all reagents used in original reference. Blocking Protocol 1. Incubate sections 1 hour at 37?C waterbath. You do no have to prewarm buffer mixed with reagents before immersing slides. 2. Rinse in DPBS 3 changes/5 minutes each 3. Proceed with immunostaining Preweigh and freeze down aliquots of glucose oxidase calculated for 100 mls working buffer, bring out, and dissolve in DPBS/sodium azide buffer, immerse slides, incubate, rinse, etc. From kmerriam2003 <@t> yahoo.com Wed Sep 15 09:17:31 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] offtopic: histologist on "the Benefactor" In-Reply-To: Message-ID: <20040915141731.37224.qmail@web52510.mail.yahoo.com> It is a TV show, I think it was on ABC. Some rich guy is giving away 1 million dollars to the winner of the show. He kicked off 3 people within a couple of hours of their arrival in the house! The histologist did not really explain what he does, but I did hear him mention autopsies to someone. Kim "Bartlett, Jeanine" wrote: Is that a TV show? What channel? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Merriam Sent: Wednesday, September 15, 2004 7:52 AM To: Histonet Subject: [Histonet] offtopic: histologist on "the Benefactor" Hello all, I know most of you are getting ready for NSH. But did anyone watch "the Benefactor"? One of the contestants is a Histologist from Portland, OR! How cool is that! Kim Merriam Novartis Cambridge, MA Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Yahoo! Mail is new and improved - Check it out! From Stephen.Eyres <@t> sanofi-synthelabo.com Wed Sep 15 09:24:16 2004 From: Stephen.Eyres <@t> sanofi-synthelabo.com (Stephen.Eyres@sanofi-synthelabo.com) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Histology of cell cultures Message-ID: Hi All, What methods are folks using to prepare histology preps of cell cultures? Any help would be much appreciated/ Cheers Steve ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- From Terry.Marshall <@t> rothgen.nhs.uk Wed Sep 15 10:02:53 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Sulfur Granule Stain Message-ID: Anything. If you get a sulfur granule (vanishingly unlikely) it will sit there as massive as a combined harvester, and you won't be able to "see into it" anyway. A tea strainer would likely have been better than ThinPrep:-) Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Paula Lucas [mailto:plucas@biopath.org] Sent: 15 September 2004 14:30 To: Histonet (E-mail) Subject: [Histonet] Sulfur Granule Stain The pathologist is requesting a sulfur granule stain on a peritoneal fluid. We prepared a thinprep slide for this. Could you tell me the best stain for this? Thank you, Paula Lucas Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From wasielewski.reinhard.von <@t> mh-hannover.de Wed Sep 15 10:11:04 2004 From: wasielewski.reinhard.von <@t> mh-hannover.de (wasielewski.reinhard.von@mh-hannover.de) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] PDGF-R in FFPE human tissue Message-ID: <414877A8.25940.95348516@localhost> Hi Histonetters, here is my next problem (maybe you have fun in solving it): we want to reproducibly stain PDGF-Receptor alpha in FFPE from human tumors. We have tried the R&D clone - but it didn't work too well. Is there anybody out there who knows better clones or secret tricks to help us to succeed ? Many thanks in advance. Reinhard PD Dr. med. Reinhard von Wasielewski From carl.hobbs <@t> kcl.ac.uk Wed Sep 15 10:13:14 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] re" Perferred" fixative for animals Message-ID: <006601c49b36$863d4c80$e8345c9f@Carlos> In my opinion, to date, no other such fixing fluid exists......OK, I havn't tried any of the commercial preps but if they were superior to formalin we'd all be using it/them. I've gone thro' the range of standard fixing fluids and also such as "Perfix" and "Sera's" fluid. None of them are superior to Formalin regarding their preservation of antigenicity/ their capacity for antigen retrieval. Some are equal for particular antigens eg Perfix -fixed mouse tissue will demonstrate phosphoHistones beautifully but not any other Ag that I'm interested in, but formalin - fixed tissue will also demonstrate them, after AR; it will also allow for the demonstration of many, many other Ags in the same block. This applies to mouse, rat, dogfish, trout, fruitfly and chick, in my experience. Why, may I ask, do you not wish to use Formalin/paraformaldehyde? From kmerriam2003 <@t> yahoo.com Wed Sep 15 10:23:29 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:03 2005 Subject: Fwd: Re: [Histonet] offtopic: histologist on "the Benefactor" Message-ID: <20040915152329.66083.qmail@web52510.mail.yahoo.com> link to bio on "the Benefactor" web site. Not sure what a traveling histologist does. Thanks for the link Diane! Note: forwarded message attached. Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Yahoo! Mail - You care about security. So do we. From lfaucette <@t> niaid.nih.gov Wed Sep 15 10:29:39 2004 From: lfaucette <@t> niaid.nih.gov (Faucette, Lawrence (NIH/NIAID)) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Histology of cell cultures Message-ID: <78F91CA4B8F3504E8FE5F13AAF122ED804A02D2A@nihexchange27.nih.gov> This is the protocol we have been using for years. Hope it helps Live cell preparation. Prepare 1% Agarose, cell culture grade, in iso osmotic PBS. You need to boil to dissolve it in PBS. Check for loss of water as vapor during boiling and replace with distilled water. Bring to approx 50 C?. The agar will solidify below this point. Prepare your cells as a cell suspension in minimal amount of medium, at room temperature. To make a 0.5 ml agar block you may need between 10 to 20x106 cells. You can scale this down to as little as 0.5 x 106 in 50 ?l but your cells will be very sparse on section. Prepare in advance 1.5 ml Eppendorf tubes (or small PCR tubes for micro-preparations) to which you cut off and discarded the conical bottom. Cap the tube. Mix evenly and thoroughly your cells with 0.5 ml of agar (or less) and very quickly transfer to the capped inverted tube. No bubbles. Place the still molten agar and tube on ice until is solid. Then open carefully the cap, and gently extract the agar cylinder from its base (the cap). At this point you can cut the agar piece in two with a razor blade, freeze half and fix in formalin the other. Your cells are still alive and biochemically active at this point. Fixed cell preparation. Proceed as above, but you fix the cells before. Then you can use Agarose in PBS, regardless of osmolarity. Fixing the cells before embedding, prevents movement of labile antigens or loss of short-lived molecules. Lawrence J Faucette Contractor HT ASCP http://www.niaid.nih.gov/dir/services/animalcare/VetPathology/VetPathology-i ndex.html Infectious Disease Pathogenesis Section Comparative Medicine Branch Division of Intramural Research, NIAID, NIH Twinbrook III, Room 2W-01A, MSC 8135 12735 Twinbrook Parkway Bethesda, MD 20892-8135 Telephone 301-451-1056 Fax 301-480-2343 SoBran, Inc. 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. The National Institute of Allergy and Infectious Diseases (NIAID) shall not accept liability for any statement made that are the sender's own and not expressly made on behalf of the NIAID by one of its representatives. -----Original Message----- From: Stephen.Eyres@sanofi-synthelabo.com [mailto:Stephen.Eyres@sanofi-synthelabo.com] Sent: Wednesday, September 15, 2004 10:24 AM To: Histonet Subject: [Histonet] Histology of cell cultures Hi All, What methods are folks using to prepare histology preps of cell cultures? Any help would be much appreciated/ Cheers Steve ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Dave.Pizi <@t> carolinashealthcare.org Wed Sep 15 10:31:35 2004 From: Dave.Pizi <@t> carolinashealthcare.org (Pizi, Dave) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Block retention Message-ID: <7E22BEDC45D51449B847E15983E99BAD0191E987@dcr-xchg-04.Carolinas.org> I hope this doesn't open a can of worms, but our medical director would like to know how other labs are handling block and slide disposal when there is a cancer center involved. His concern is that even though we are following CAP and state guidelines when we discard after 10 years, the cancer center folks will request something older and we won't be able to help them. I can't think of a way to identify, retrieve and hold this material that doesn't give me a headache. Thanks Dave Pizi Carolinas HealthCare System Charlotte, NC ----------------------------------------- This electronic message may contain information that is confidential and/or legally privileged. It is intended only for the use of the individual(s) and entity named as recipients in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from any computer. Do not deliver, distribute or copy this message, and do not disclose its contents or take any action in reliance on the information it contains. Thank you. From JWEEMS <@t> sjha.org Wed Sep 15 10:40:04 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Block retention Message-ID: <83AACDB0810528418AA106F9AE9B7F7E2782D2@sjhaexc02.sjha.org> Same concern here - I almost had a coronory when I had to tell a patient we had discarded her case per regulations and she said "I just wasn't supposed to live this long". I hope to see this reviewed. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Pizi, Dave Sent: Wednesday, September 15, 2004 11:32 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Block retention I hope this doesn't open a can of worms, but our medical director would like to know how other labs are handling block and slide disposal when there is a cancer center involved. His concern is that even though we are following CAP and state guidelines when we discard after 10 years, the cancer center folks will request something older and we won't be able to help them. I can't think of a way to identify, retrieve and hold this material that doesn't give me a headache. Thanks Dave Pizi Carolinas HealthCare System Charlotte, NC ----------------------------------------- This electronic message may contain information that is confidential and/or legally privileged. It is intended only for the use of the individual(s) and entity named as recipients in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from any computer. Do not deliver, distribute or copy this message, and do not disclose its contents or take any action in reliance on the information it contains. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From ander093 <@t> tc.umn.edu Wed Sep 15 10:56:48 2004 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Block retention In-Reply-To: <7E22BEDC45D51449B847E15983E99BAD0191E987@dcr-xchg-04.Carol inas.org> Message-ID: <5.2.0.9.0.20040915105049.00a3eec0@ander093.email.umn.edu> We have all slides and blocks dating back at least 30 years or more. We have received many requests over the years for slides that are older than 10 years--so this is a very valid concern, IMHO. We have a separate store room where these slides and blocks are kept. Our Neuropathologist does not want anything discarded--don't know what we'll do when the storeroom fills up--but we will be able to pull slides/blocks if needed. I do agree that this policy should be reviewed. LuAnn At 11:31 AM 9/15/04 -0400, Pizi, Dave wrote: >I hope this doesn't open a can of worms, but our medical director would >like to know how other labs are handling block and slide disposal when >there is a cancer center involved. His concern is that even though we are >following CAP and state guidelines when we discard after 10 years, the >cancer center folks will request something older and we won't be able to >help them. I can't think of a way to identify, retrieve and hold this >material that doesn't give me a headache. > >Thanks >Dave Pizi >Carolinas HealthCare System >Charlotte, NC > >----------------------------------------- >This electronic message may contain information that is confidential >and/or legally privileged. It is intended only for the use of the >individual(s) and entity named as recipients in the message. If you are >not an intended recipient of this message, please notify the sender >immediately and delete the material from any computer. Do not deliver, >distribute or copy this message, and do not disclose its contents or take >any action in reliance on the information it contains. Thank you. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jill.Cox <@t> providence.org Wed Sep 15 11:09:04 2004 From: Jill.Cox <@t> providence.org (Cox, Jill) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Cutting controls Message-ID: <05424658AAA6D441BBAFD9D9D98780B7044F0CB1@lcmmsg02.ca.providence.org> Hello Netters, I had previously posted a message on starting a control cutting business. Well I did it!!! Thank you for all your help. My website is the only thing I have left to finish. If anyone needs any assistance in control slides or just cutting blocks I would love to help you. Now you at least have one more resource. I hope this is ok content to post. Thanks again... Jill Cox HT ascp Pathology Supervisor Little Company of Mary 4101 Torrance Blvd. Torrance Ca. 90503 (310) 543-6736 DISCLAIMER: This message is intended for the sole use of the addressee, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If you are not the addressee you are hereby notified that you may not use, copy, disclose, or distribute to anyone the message or any information contained in the message. If you have received this message in error, please immediately advise the sender by reply email and delete this message. From bill501 <@t> mindspring.com Wed Sep 15 11:27:02 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Sulfur Granule Stain In-Reply-To: References: Message-ID: At 4:02 PM +0100 9/15/04, Marshall Terry Dr, Consultant Histopathologist wrote: >A tea strainer would likely have been better than ThinPrep:-) LOL! I do use tea strainers in the gross room (American tea strainers...). Almost anything will stain them - an H&E is fine. They are PAS+, AFB+ Gram+ etc It might be better to culture for actinomyces... If this is a serious possibility As I recall... BB -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board From jluis.palazon <@t> icman.csic.es Wed Sep 15 11:31:57 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Gomori chrome alum hematoxylin Message-ID: <20040915163157.544262C3A2A@perceval.uca.es> Dear list-members Greetings. I can not find the recipe to prepare Gomori?s chrome alum hematoxylin. Could any of you help me please? Thanks in advance Jos? Luis From failm <@t> musc.edu Wed Sep 15 11:56:25 2004 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] CD179 Message-ID: Has anyone used this marker? IF so at what dilution Thank Rena Fail From nsdnz <@t> neuro.hfh.edu Wed Sep 15 12:54:36 2004 From: nsdnz <@t> neuro.hfh.edu (Danielle Zalinski) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] CD31/PECAM in Rat Brain Message-ID: <414881DC.6070502@neuro.hfh.edu> Hello Histonetters, Our lab is interested in looking at CD31/PECAM in rat brain tissue as a marker for neovessels. We don't know anyone in our facility who has used this antibody extensively, so we are at a loss for deciding on a manufacturer. Has anyone worked with this antibody and has a (successful) protocol and manufacturer that they prefer? We would prefer a mouse monoclonal for double labeling, but polyclonal is ok, too. Does anyone know if we will need to use any exotic fixation/retrieval techniques? We want to be able to work out a protocol first in paraffin and may adapt to frozen sections later. Thank you for any help you can provide, and if you need more info, please let me know, Danielle Zalinski Henry Ford Health System Detroit, MI CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by replying in an email to the sender, delete the email from your computer system and shred any paper copies of the email you printed. Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. These risks are described in our Privacy Policy at http://henryford.com. Review that policy carefully before continuing to communicate with us by e-mail. For greater Internet security, our policy describes the Henry Ford MyHealth electronic communication process - you may register at http://henryford.com. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. From bryand <@t> netbistro.com Wed Sep 15 13:20:22 2004 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Gomori chrome alum hematoxylin In-Reply-To: <20040915163157.544262C3A2A@perceval.uca.es> References: <20040915163157.544262C3A2A@perceval.uca.es> Message-ID: <414887E6.7040609@netbistro.com> According to Gray it is:- Water - 100 mL Sulphuric acid, 5% aqu. - 2 mL Potassium dichromate 5% aqu. - 2 mL Chrome alum - 1.5 g Hematoxylin - 0.5 g Dissolve the alum and the hematoxylin each in 50 mL water. Combine the two solutions. Add the potassium dichromate and sulphuric acid. Leave for 48 hours to ripen. Bryan Llewellyn Jose Luis Palazon Fernandez wrote: > Dear list-members > > > > Greetings. I can not find the recipe to prepare Gomori?s chrome alum hematoxylin. Could any of you help me please? > > > > Thanks in advance > > > > > > Jos? Luis > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From DEllenburg2 <@t> stfrancishealth.org Wed Sep 15 14:18:26 2004 From: DEllenburg2 <@t> stfrancishealth.org (DEllenburg2@stfrancishealth.org) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] RE: Histonet Digest, Vol 10, Issue 22 Message-ID: <6661F19BB774D711942900A0C905F2533D8BD8@GVL01MSX> Not sure if this is what you are looking far. CHROMIUM HEMATOXYLIN SOLUTION. Hematoxylin, 1% aqueous solution................................ 50.0 cc. Chromium alum, 3% aqueous solution.............................. 50.0 cc. To 100 cc. of chromium hematoxylin add 0.1 gm. of potassium iodate. Boil until a deep blue. The mixture is ripe immediately anc can be used as long as a film with a metallic luster forms on its surface in a Coplin jar. Filter before use. Hope this helps. If you would like I will fax this procedure to you. Deborah Ellenburg, HT (ASCP) Histology Supervisor Bon Secours St. Francis Health System (phone) 864-255-1278 (fax) 864-2551664 -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: None To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 10, Issue 22 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Re: Block retention (LuAnn Anderson) 2. Cutting controls (Cox, Jill) 3. RE: Sulfur Granule Stain (Bill Blank) 4. Gomori chrome alum hematoxylin (Jose Luis Palazon Fernandez) 5. CD179 (Mildred Fail) ---------------------------------------------------------------------- Message: 1 Date: Wed, 15 Sep 2004 10:56:48 -0500 From: LuAnn Anderson Subject: Re: [Histonet] Block retention To: "Pizi, Dave" , Message-ID: <5.2.0.9.0.20040915105049.00a3eec0@ander093.email.umn.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed We have all slides and blocks dating back at least 30 years or more. We have received many requests over the years for slides that are older than 10 years--so this is a very valid concern, IMHO. We have a separate store room where these slides and blocks are kept. Our Neuropathologist does not want anything discarded--don't know what we'll do when the storeroom fills up--but we will be able to pull slides/blocks if needed. I do agree that this policy should be reviewed. LuAnn At 11:31 AM 9/15/04 -0400, Pizi, Dave wrote: >I hope this doesn't open a can of worms, but our medical director would >like to know how other labs are handling block and slide disposal when >there is a cancer center involved. His concern is that even though we are >following CAP and state guidelines when we discard after 10 years, the >cancer center folks will request something older and we won't be able to >help them. I can't think of a way to identify, retrieve and hold this >material that doesn't give me a headache. > >Thanks >Dave Pizi >Carolinas HealthCare System >Charlotte, NC > >----------------------------------------- >This electronic message may contain information that is confidential >and/or legally privileged. It is intended only for the use of the >individual(s) and entity named as recipients in the message. If you are >not an intended recipient of this message, please notify the sender >immediately and delete the material from any computer. Do not deliver, >distribute or copy this message, and do not disclose its contents or take >any action in reliance on the information it contains. Thank you. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 2 Date: Wed, 15 Sep 2004 09:09:04 -0700 From: "Cox, Jill" Subject: [Histonet] Cutting controls To: Message-ID: <05424658AAA6D441BBAFD9D9D98780B7044F0CB1@lcmmsg02.ca.providence.org> Content-Type: text/plain; charset="Windows-1252" Hello Netters, I had previously posted a message on starting a control cutting business. Well I did it!!! Thank you for all your help. My website is the only thing I have left to finish. If anyone needs any assistance in control slides or just cutting blocks I would love to help you. Now you at least have one more resource. I hope this is ok content to post. Thanks again... Jill Cox HT ascp Pathology Supervisor Little Company of Mary 4101 Torrance Blvd. Torrance Ca. 90503 (310) 543-6736 DISCLAIMER: This message is intended for the sole use of the addressee, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. If you are not the addressee you are hereby notified that you may not use, copy, disclose, or distribute to anyone the message or any information contained in the message. If you have received this message in error, please immediately advise the sender by reply email and delete this message. ------------------------------ Message: 3 Date: Wed, 15 Sep 2004 11:27:02 -0500 From: Bill Blank Subject: RE: [Histonet] Sulfur Granule Stain To: "Marshall Terry Dr, Consultant Histopathologist" , "Paula Lucas" , "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" At 4:02 PM +0100 9/15/04, Marshall Terry Dr, Consultant Histopathologist wrote: >A tea strainer would likely have been better than ThinPrep:-) LOL! I do use tea strainers in the gross room (American tea strainers...). Almost anything will stain them - an H&E is fine. They are PAS+, AFB+ Gram+ etc It might be better to culture for actinomyces... If this is a serious possibility As I recall... BB -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board ------------------------------ Message: 4 Date: Wed, 15 Sep 2004 18:31:57 +0200 (CEST) From: Jose Luis Palazon Fernandez Subject: [Histonet] Gomori chrome alum hematoxylin To: histonet@lists.utsouthwestern.edu Message-ID: <20040915163157.544262C3A2A@perceval.uca.es> Content-Type: text/plain; charset="iso-8859-1" Dear list-members Greetings. I can not find the recipe to prepare Gomori?s chrome alum hematoxylin. Could any of you help me please? Thanks in advance Jos? Luis ------------------------------ Message: 5 Date: Wed, 15 Sep 2004 12:56:25 -0400 From: "Mildred Fail" Subject: [Histonet] CD179 To: Message-ID: Content-Type: text/plain; charset=US-ASCII Has anyone used this marker? IF so at what dilution Thank Rena Fail ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 10, Issue 22 **************************************** The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at (864) 255-1000, and permanently delete the original e-mail, attachment(s), and any copies. From froyer <@t> bitstream.net Wed Sep 15 14:27:21 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:03 2005 Subject: Fwd: Re: [Histonet] offtopic: histologist on "the Benefactor" In-Reply-To: <20040915152329.66083.qmail@web52510.mail.yahoo.com> References: <20040915152329.66083.qmail@web52510.mail.yahoo.com> Message-ID: <41489799.8010901@bitstream.net> Kim Merriam wrote: >link to bio on "the Benefactor" web site. Not sure what a traveling histologist does. > > ....they gather no moss? maybe? ~ Ford Ford M. Royer M.S.B. Minneapolis, MN From John.Garlits <@t> STJUDE.ORG Wed Sep 15 14:33:28 2004 From: John.Garlits <@t> STJUDE.ORG (Garlits, John) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] EDTA decal protocol? Message-ID: <1E0CC447E59C974CA5C7160D2A2854EC03C40E9C@SJMEMXMB04.stjude.sjcrh.local> Hello, I am looking for a well-established EDTA decalcification protocol to use on mouse bones for IHC. Could anyone who does this let me know what reagents and times they use? Thanks! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 Phone: 901-495-3961 Fax: 901-495-2176 From funderwood <@t> mcohio.org Wed Sep 15 14:31:23 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:03 2005 Subject: [BULK] - Re: Fwd: Re: [Histonet] offtopic: histologist on "the Benefactor" Message-ID: I have that CD. It has Tom Petty, Jeff Lynne, George Harrison, Bob Dylan and Roy Orbison. >>> Ford Royer 09/15/04 03:27PM >>> Kim Merriam wrote: >link to bio on "the Benefactor" web site. Not sure what a traveling histologist does. > > ....they gather no moss? maybe? ~ Ford Ford M. Royer M.S.B. Minneapolis, MN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From froyer <@t> bitstream.net Wed Sep 15 16:11:53 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:03 2005 Subject: [BULK] - Re: Fwd: Re: [Histonet] offtopic: histologist on "the Benefactor" In-Reply-To: References: Message-ID: <4148B019.4030604@bitstream.net> WOW!! I never knew they were Histologist!! Fred Underwood wrote: >I have that CD. It has Tom Petty, Jeff Lynne, George Harrison, Bob >Dylan and Roy Orbison. > > > >>>>Ford Royer 09/15/04 03:27PM >>> >>>> >>>> >Kim Merriam wrote: > > > >>link to bio on "the Benefactor" web site. Not sure what a traveling >> >> >histologist does. > > >> >> >> >> >....they gather no moss? maybe? > >~ Ford >Ford M. Royer >M.S.B. >Minneapolis, MN > >_______________________________________________ >Histonet mailing list >Histonet@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 heytallguy2000 <@t> hotmail.com Wed Sep 15 18:29:04 2004 From: heytallguy2000 <@t> hotmail.com (john clark) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] microwave processors Message-ID: Our lab needs a new processor and while we are looking we are interesting in microwave processing for our bx. Has anyone used a processor that they have been satisfied with the quality and labor intensity? The Sakura microwave processor is out of our buget. I would appreciate any suggestions. John From JAnorthernexp <@t> cs.com Wed Sep 15 18:50:41 2004 From: JAnorthernexp <@t> cs.com (JAnorthernexp@cs.com) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Re: Double antibody staining Message-ID: <1c1.1e446edb.2e7a2f51@cs.com> Does anyone out there know if Ventana's Nexus stainer does double antibody staining? Annette C Hart, HT(ASCP) Pennant Labs Wilkes-Barre, PA From bhewlett <@t> cogeco.ca Wed Sep 15 19:05:23 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] re" Perferred" fixative for animals References: <006601c49b36$863d4c80$e8345c9f@Carlos> Message-ID: <003901c49b80$ddf72fa0$6400a8c0@mainbox> Couldn't have said it better myself! Amen, Carl. Bryan ----- Original Message ----- From: "Carl Hobbs" To: Sent: Wednesday, September 15, 2004 11:13 AM Subject: [Histonet] re" Perferred" fixative for animals > In my opinion, to date, no other such fixing fluid exists......OK, I havn't > tried any of the commercial preps but if they were superior to formalin > we'd all be using it/them. I've gone thro' the range of standard fixing > fluids and also such as "Perfix" and "Sera's" fluid. None of them are > superior to Formalin regarding their preservation of antigenicity/ their > capacity for antigen retrieval. Some are equal for particular antigens eg > Perfix -fixed mouse tissue will demonstrate phosphoHistones beautifully but > not any other Ag that I'm interested in, but formalin - fixed tissue will > also demonstrate them, after AR; it will also allow for the demonstration > of many, many other Ags in the same block. This applies to mouse, rat, > dogfish, trout, fruitfly and chick, in my experience. > Why, may I ask, do you not wish to use Formalin/paraformaldehyde? > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From vandries <@t> vub.ac.be Thu Sep 16 01:59:17 2004 From: vandries <@t> vub.ac.be (Veronique Andriessen) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Glucose oxidase In-Reply-To: <20040915160024.61474.qmail@web50309.mail.yahoo.com> Message-ID: Thanks for your suggestions Phil, but I've tried all that. I made the solution fresh and added the GLUOX to the buffer just before putting it in the 37C water bath. I also tried omitting NaN3 and different concentrations of GLUOX in the buffer but nothing worked. Veronique -----Original Message----- From: Phillip Huff [mailto:histophilhuff@yahoo.com] Sent: woensdag 15 september 2004 18:00 To: Veronique Andriessen Subject: Re: [Histonet] Glucose oxidase The glucose oxidase procedure requires that the solution is fresh when used. We make the solution immediately before we need it, warming it in a water bath (37C) for 5 minutes before use. We currently do not use Sodium Azide in our reactions so you could try dropping that reagent and make the solution fresh. Good luck, Phil Veronique Andriessen wrote: Hi histonetters, I've got a problem. I'm have to do immunostaining with HRP and AP on inflamed rat pancreas (fresh frozen sections, fixed with acetone ethanol 3:1). I have to quench the endogenous peroxidase in the white blood cells. I have tried the glucose oxidase block as described in the histonet archives, but I am not able to get it working. I have ordered the correct ingredients from Sigma, but nothing! The only thing that is not recently purchased is the Sodium azide, but I can't imagine this going bad. I even tried adding 0.1% saponin, but this also did't help. I really need some advice, I'm getting desparate. Kindest regards, Veronique Andriessen BAS Lab. Molecular Liver Cell Biology Free University Brussels (VUB), Belgium Tel: 0032-2-477.4259 Fax: 0032-2-477.4412 vandries@vub.ac.be GLUCOSE OXIDASE BLOCK FOR ENDOGENOUS PEROXIDASE/PSEUDOPEORXIDASES IN FROZEN/PARAFFIN SECTIONS Reference: Andrew SM, Jasani B. An improved method for the inhibition of endogenous peroxidase non-deleterious to lymphocyte surface markers. Application to immunoperoxidase studies on eosinophil-rich tissue preparations. Histochemical Journal 19:426-30, 1987 All forms of endogenous peroxidase may not be inhibited by the usual methanol/hydrogen peroxide or buffer/hydrogen peroxide blocking mixtures. This method produces nascent hydrogen peroxide that is preferable to the normal methods using preformed hydrogen peroxide (which you add or buy in your endogenous peroxide blocker). This new method to block 'peroxidatic activity' is consistently complete. This method is an enzymatic reaction that produces a slow, steady, very low concentration of hydrogen peroxide. It is particularly usefu l for monoclonal antibody staining on frozen sections that are minimally fixed with acetone. Procedure: Glucose (Sigma G 5250) 0.180 g Glucose oxidase (Sigma G 6641) 0.005 g Sodium azide 0.0065g DPBS 50 ml This procedure has doubled concentration of all reagents used in original reference. Blocking Protocol 1. Incubate sections 1 hour at 37?C waterbath. You do no have to prewarm buffer mixed with reagents before immersing slides. 2. Rinse in DPBS 3 changes/5 minutes each 3. Proceed with immunostaining Preweigh and freeze down aliquots of glucose oxidase calculated for 100 mls working buffer, bring out, and dissolve in DPBS/sodium azide buffer, immerse slides, incubate, rinse, etc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------------- -- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. From joeamateur <@t> hotmail.com Wed Sep 15 13:54:40 2004 From: joeamateur <@t> hotmail.com (Jack England) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] vascular graft_sectioning difficulty Message-ID: Can you provide a little more information about your processing, and a little more detail about how the sections are crumbling (eg, tissue is disaggregating, or the paraffin section is falling apart, etc)? We regularly section formalin-fixed, paraffin-embedded vascular grafts (ePTFE) at 5-7µm, and it's working for us. With a little more information, I might be able to help... Aloha, Jack England Tissue Genesis, Inc. http://www.tissuegenesis.com >From: "Dr. Raghul" >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] vascular graft_sectioning difficulty >Date: Tue, 14 Sep 2004 16:56:23 +0530 > >hello members, > >We cut 5 µm thin sections of paraffin embedded vascular graft(Dacron) in >a rotary microtome with disposable blades but the senior tech tells that >the tissue sections are crumbling and forces us to go for thick sections >around 10µm but again not with much success. > > We can understand that there is not enough tissue adhereing to the >graft and we are literally cutting the graft. But did anyone had such >problems sectioning vascular grafts. Any suggestions? > > >regards , > >Raghul J >scientist >Histopathology- implant biology division >Srichitra tirunal institute of medical sciences and technology >Trivandrum -695 012 >Kerala, India > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar – get it now! http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/ From raghul <@t> sctimst.ker.nic.in Thu Sep 16 06:32:16 2004 From: raghul <@t> sctimst.ker.nic.in (Dr. Raghul) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] vascular graft_sectioning difficulty In-Reply-To: References: Message-ID: Hello Aloha, Our vascular graft material(DACRON) after fixing in 10% NBF, we dehydrate in ascending grades of alcohol(70,80,95,100%),cleared in 3 changes of chloroform before embedding in paraffin wax (60*C MPT). While cutting, the graft area is powdering in the sections obtained.When it comes to the anastomising graft-host vessel part, while the vessel portion comes well, the graft as usual gets powdered . This is our processing detail and the sectioning difficulty we face.Still if the details are insufficient please ask. regards raghul -----Original Message----- From: "Jack England" To: raghul@sctimst.ker.nic.in, Histonet@lists.utsouthwestern.edu Date: Wed, 15 Sep 2004 08:54:40 -1000 Subject: RE: [Histonet] vascular graft_sectioning difficulty > Can you provide a little more information about your processing, and a > little more detail about how the sections are crumbling (eg, tissue is > disaggregating, or the paraffin section is falling apart, etc)? We > regularly > section formalin-fixed, paraffin-embedded vascular grafts (ePTFE) at > 5-7?m, > and it's working for us. With a little more information, I might be > able to > help... > > Aloha, > Jack England > Tissue Genesis, Inc. > http://www.tissuegenesis.com > > >From: "Dr. Raghul" > >To: histonet@lists.utsouthwestern.edu > >Subject: [Histonet] vascular graft_sectioning difficulty > >Date: Tue, 14 Sep 2004 16:56:23 +0530 > > > >hello members, > > > >We cut 5 ?m thin sections of paraffin embedded vascular graft(Dacron) > in > >a rotary microtome with disposable blades but the senior tech tells > that > >the tissue sections are crumbling and forces us to go for thick > sections > >around 10?m but again not with much success. > > > > We can understand that there is not enough tissue adhereing to the > >graft and we are literally cutting the graft. But did anyone had such > >problems sectioning vascular grafts. Any suggestions? > > > > > >regards , > > > >Raghul J > >scientist > >Histopathology- implant biology division > >Srichitra tirunal institute of medical sciences and technology > >Trivandrum -695 012 > >Kerala, India > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _________________________________________________________________ > FREE pop-up blocking with the new MSN Toolbar ? get it now! > http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/ From MAUGER <@t> email.chop.edu Thu Sep 16 07:19:02 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] CD31/PECAM in Rat Brain Message-ID: To Danielle, Pharmingen has a good rat anti ms CD31 catalog# 557355. It works great on ms tissue with pepsin digestion. I use a rabbit anti rat secondary(ms adsorbed)from Vector. Not sure how it will work on rat. Jo >>> "Danielle Zalinski" 09/15/04 01:54PM >>> Hello Histonetters, Our lab is interested in looking at CD31/PECAM in rat brain tissue as a marker for neovessels. We don't know anyone in our facility who has used this antibody extensively, so we are at a loss for deciding on a manufacturer. Has anyone worked with this antibody and has a (successful) protocol and manufacturer that they prefer? We would prefer a mouse monoclonal for double labeling, but polyclonal is ok, too. Does anyone know if we will need to use any exotic fixation/retrieval techniques? We want to be able to work out a protocol first in paraffin and may adapt to frozen sections later. Thank you for any help you can provide, and if you need more info, please let me know, Danielle Zalinski Henry Ford Health System Detroit, MI CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by replying in an email to the sender, delete the email from your computer system and shred any paper copies of the email you printed. Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. These risks are described in our Privacy Policy at http://henryford.com. Review that policy carefully before continuing to communicate with us by e-mail. For greater Internet security, our policy describes the Henry Ford MyHealth electronic communication process - you may register at http://henryford.com. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Thu Sep 16 09:25:52 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Fixation of bone marrow smears for IHC Message-ID: Help. I would appreciate some input on fixation of murine bone marrow smears for future IHC. We're harvesting smears and femurs this afternoon, and then I'm going on vacation. Of course the world stops spinning. I will potentially stain these smears for B + T cells in a couple of weeks, and possibly platelets, like CD61. The bones will be handled in my absence, but I'm worried about how to fix and store the smears. Anyone with experience with IHC on bone marrow smears will be handsomely rewarded. (Not really, but I'll take whatever I can get.) Thanks! My daughter is getting married in a week, and my brain isn't working right. Jackie O' From anaflorenti <@t> yahoo.com Thu Sep 16 09:56:28 2004 From: anaflorenti <@t> yahoo.com (Ana Florentin) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] endogenous peroxidase blocking protocol Message-ID: <20040916145628.72162.qmail@web52609.mail.yahoo.com> Hi, Does anybody know a good protocol for blocking the endogenous peroxidase? I am performing immunoperoxidase staining on rat spleen frozen sections and I have tried to block endogenous peroxidase with H2O2, 1% and 0.3% in PBS, half hour before the first antibody but is seemed to be a too high concentration, as the tissue was destroyed. Thanks for help! anne __________________________________ Do you Yahoo!? New and Improved Yahoo! Mail - 100MB free storage! http://promotions.yahoo.com/new_mail From pzeitlow <@t> bbpllab.com Thu Sep 16 11:53:00 2004 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Pathvysion Her2/neu Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A0BEA73@bbplsrv1.bbpl> I have some specific questions regarding findings with Pathvysion Her2. Is anyone doing these that would be willing to share and compare findings? Pat Z From TJasper <@t> smdc.org Thu Sep 16 12:44:44 2004 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] fixed and unfixed immuno and antuibody oreps Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E44FB0@harrier> Dear George, I have read some of your postings and I have developed a soft spot for you. It seems what you're after is an explanation of how immunohistochemistry can be done these days using techniques which negated your efforts in the past. Also you seem to be questioning how the "best" method was determined, why is it OK to do IHC the way most labs do? George, I honestly cannot answer how today's techniques were exactly determined/developed. I am not an expert in the field of IHC history and comparative research of the same. Here's what I do know, I have been doing Histology work for approximately 20 years (I know a babe in woods). All of my training and experience in doing IHC has been heavily dependent upon proper fixation. This is for the most part formalin fixed (10% Neutral Buffered), paraffin processed tissue specimens. Most people working in the clinical world, as I do, base the majority of their IHC work on properly fixed tissue specimens. This is how we understand IHC. My lab runs into problems occasionally when we receive referral cases (either block or slides) that have poor/improper fixation. This hinders our ability to produce a good quality diagnostic stain. Antigen retrieval techniques are employed to enhance staining and maybe that is the answer you seek as it is a post-fixation procedure. George, I am not a researcher and there are a lot of people on the histonet that are better qualified than I am to speak to the finer points of IHC. I just thought I would share my thoughts with you. I consider myself to be a standard histologist with a fairly basic understanding of IHC as it relates to I what I am expected to put out on a daily basis. George, I apologize if the things I have just stated have already been related to you or if someone with a greater understanding of IHC finds flaws in what I've posted. Being a manager I don't do quite as much IHC as I used to, although I certainly have my hand in the day-to-day immunos that are run here. I also sympathize with you in a general sense as I do like to understand things and like clear explanations and/or reasons why. Hope this helps. Regards, Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: George Cole [mailto:georgecole@ev1.net] Sent: Tuesday, September 14, 2004 1:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fixed and unfixed immuno and antuibody oreps I promise to drop this line---I have had an itch to find out something--- I realize I must give it up---I have asked it repeatedly and I don't want to keep asking the same thing---impatience is bound to develop. Retired as I am---I no longer have lab in which to answer such questions. Most histotechs nowadays are brought up doing one side of the question---indeed it is the standard procedure and my question just doesn't scan with today's techs. But if I could shuck 10 years and be back in my lab and I was given the responsibility to do a certain class of work I would do the following: Antibody technique or immunofluorescence procedure---any procedure that has a fixative in its beginning followed by some procedure to neutralize the effects of the fixative----I would run at least 3 trials on the 2 sets of pairs: 1) Tissue unfixed 2) tissue fixed, then processed with by whatever step was supposed to neutralize the fixation effect, rinsing the tissues carefully afterwards. . Then I would run tissues 1 and 2 in the same preparation. And I would choose as my standard procedure which ever procedure of the pair which gave the better results. I drop that and run. The crowding effect of Standard Procedure Itis is out there. But then I might just find my question was trivial and standard procedure might win. But has the fix-unfix procedure ever been tested against the unfixed prep? This retired picky-compulsive tech would never run ANY procedure as standard without testing its effectiveness---comparing it--- with any reasonable alternative procedure for quality. Nuff said. Retired Tech now enters a stage of quiet upon the subject on the histonet, but I will still brood over the untapped quality test in ANY procedure done by rote rather than via the search for the most excellent way to do that test. georgecole@ev1.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sluhisto <@t> yahoo.com Thu Sep 16 12:48:27 2004 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] CD3: CD23 Message-ID: <20040916174827.58880.qmail@web51003.mail.yahoo.com> Hello All: Is anyone using CD5 and CD23 on B5 fixed tissue? I need to know manufacturer, dilution, retrieval, system, etc. We are having problems with these on B5 fixed tissue. Any help is appreciated. Thanks. Susan --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From Vickroy.Jim <@t> mhsil.com Thu Sep 16 13:24:48 2004 From: Vickroy.Jim <@t> mhsil.com (Vickroy, Jim) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] job listing for Springfield, Illinois Message-ID: Memorial Medical Center Springfield, IL Histotechnician, Full-Time Days Memorial Health System is a multi-hospital integrated healthcare delivery system that includes primary care practices and home health services. We are one of the leading community-based, not-for-profit healthcare organizations in Illinois that is dedicated to patient care, education and research. Memorial Medical Center, the flagship of MHS, which is located in Springfield, is a teaching hospital affiliated with Southern Illinois University School of Medicine. As one of the state's leading healthcare providers Memorial Health System depends on a unified team effort to deliver superior services in every department at every facility. That's why it's so important to us to hire conscientious individuals who take pride in a job well done, and share our commitment to excellence. "Are you one of them?" If so, we invite you to join our team. Currently, Memorial Medical Center is seeking a dynamic individual to join our Histology Department. The selected candidate will process tissues and prepare slides for diagnosis by our pathologists. Some additional duties will include: logging, retrieving and charging for procedures; process surgical, autopsy, and SIU research material; interpret reactions, results and protocols to assess validity and check for common or unusual problems. Qualifications for the position include: high school graduate required, 1 year training at an approved school of histology and HT (ASCP) registered or registry eligible. We offer competitive compensation and comprehensive benefit packages, which includes a variety of medical/dental plans, tax deferred annuities, pension plan, life and disability insurance, and tuition reimbursement. Qualified candidates please submit a resume or inquiry to: Memorial Health System Attn: Jennifer Davis, PHR Human Resources Department 701 North First Street Springfield, Illinois 62781 217-788-3580, (Fax) 217-788-5539 davis.Jennifer@mhsil.com EOE/Drug-Free Workplace WEBISTE www.memorialmedical.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. From riccim <@t> vvc.edu Thu Sep 16 14:10:57 2004 From: riccim <@t> vvc.edu (Melody Ricci) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] CD31/PECAM Message-ID: Check out www.pharmingen.com I used to work there several years ago, and we developed IHC antibodies primarily for mouse & rat tissues. A phone call to their IHC-Quality Control department directly in San Diego will usually get you personalized help on protocols or staining problems. Good Luck M. Ricci From sluhisto <@t> yahoo.com Thu Sep 16 14:42:12 2004 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Eosinophils in mouse tissue Message-ID: <20040916194212.51844.qmail@web51004.mail.yahoo.com> Does anyone out there have a protocol for staining eosinophils in mouse lung? Thanks. Todd --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From Barry.R.Rittman <@t> uth.tmc.edu Thu Sep 16 15:15:01 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] RE:Fixed etc. long response Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0F1350A@UTHEVS3.mail.uthouston.edu> Thank you George and Thomas for raising and discussing this topic. I think that there is always somewhat of a disconnect between biologic and biochemical procedures. In biochemical work an experiment was usually carried out three times and if it worked all three times was regarded as valid. In the biologic literature some things if they work once are published as rock hard truths. Sometimes such results should be labeled "observations" and be indicators rather than hard facts but once published are usually taken as hard facts. Part of this is the tendency to examine the most attractive field rather than all fields, and it seems to me that this is especially true of electron microscopy. IHC is really following in the footsteps of histochemistry. Initially there were relatively few techniques and several positive and negative controls were always used. If enzymes were studied, their location in frozen or freeze dried sections was used as the gold standard although this was usually a cumbersome process and not always possible. Sometimes fixatives were used but the results were compared to the gold standard. As with general histology, the standard for looking at tissues was fixed tissues processed to paraffin wax. For those not used to examining frozen sections, The image in frozen sections was sometimes difficult to compare to that of fixed tissues due largely to the smaller degree of shrinkage and the more open appearance of tissues. I agree with George's general sentiment about fixation as the tissue is not in its natural state and even if antigen retrieval is used it doesn't mean that the original state has been restored. Another problems is that in some cases the claims of manufacturers regarding their antibodies has been taken as the gospel truth. One of the best examples I know concerns claims for "pan keratin" antibodies. We would assume from some of the claims that virtually all cytokeratins can be demonstrated with this antibody. In most cases nothing could be further from the truth. Before I use an antibody I would like know how it binds to epitopes, preferably in the native state. Most labs use buffered formalin, and specific times etc. However, common sense tells me that the sections produced by one lab will differ, albeit in small measure from another lab. It would therefore be logical to have a standard technique for each lab based on testing a particular antibody on their processed tissue and then comparing this to results using frozen section. Once this comparison has been made you can standardize to your procedure. Individual labs fix, process, section and stain in slightly differently. Sometimes IHC is tolerant of variation, sometimes not. It would be optimal if a standardized image was available to all labs so that results could be compared and techniques in individual labs changed to obtain this optimal result. In answer to you question, no I have not been drinking, its only 3 pm!! I know that this is pie in the sky but I am worried about the different responses that we get on Histonet regarding the same technique and often using the same antibody. Perhaps we could have a set of IHC images using fluorescence and another using peroxides so that we could get some standardization? Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jasper, Thomas G. Sent: Thursday, September 16, 2004 12:45 PM To: 'George Cole'; 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] fixed and unfixed immuno and antuibody oreps Dear George, I have read some of your postings and I have developed a soft spot for you. It seems what you're after is an explanation of how immunohistochemistry can be done these days using techniques which negated your efforts in the past. Also you seem to be questioning how the "best" method was determined, why is it OK to do IHC the way most labs do? George, I honestly cannot answer how today's techniques were exactly determined/developed. I am not an expert in the field of IHC history and comparative research of the same. Here's what I do know, I have been doing Histology work for approximately 20 years (I know a babe in woods). All of my training and experience in doing IHC has been heavily dependent upon proper fixation. This is for the most part formalin fixed (10% Neutral Buffered), paraffin processed tissue specimens. Most people working in the clinical world, as I do, base the majority of their IHC work on properly fixed tissue specimens. This is how we understand IHC. My lab runs into problems occasionally when we receive referral cases (either block or slides) that have poor/improper fixation. This hinders our ability to produce a good quality diagnostic stain. Antigen retrieval techniques are employed to enhance staining and maybe that is the answer you seek as it is a post-fixation procedure. George, I am not a researcher and there are a lot of people on the histonet that are better qualified than I am to speak to the finer points of IHC. I just thought I would share my thoughts with you. I consider myself to be a standard histologist with a fairly basic understanding of IHC as it relates to I what I am expected to put out on a daily basis. George, I apologize if the things I have just stated have already been related to you or if someone with a greater understanding of IHC finds flaws in what I've posted. Being a manager I don't do quite as much IHC as I used to, although I certainly have my hand in the day-to-day immunos that are run here. I also sympathize with you in a general sense as I do like to understand things and like clear explanations and/or reasons why. Hope this helps. Regards, Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: George Cole [mailto:georgecole@ev1.net] Sent: Tuesday, September 14, 2004 1:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fixed and unfixed immuno and antuibody oreps I promise to drop this line---I have had an itch to find out something--- I realize I must give it up---I have asked it repeatedly and I don't want to keep asking the same thing---impatience is bound to develop. Retired as I am---I no longer have lab in which to answer such questions. Most histotechs nowadays are brought up doing one side of the question---indeed it is the standard procedure and my question just doesn't scan with today's techs. But if I could shuck 10 years and be back in my lab and I was given the responsibility to do a certain class of work I would do the following: Antibody technique or immunofluorescence procedure---any procedure that has a fixative in its beginning followed by some procedure to neutralize the effects of the fixative----I would run at least 3 trials on the 2 sets of pairs: 1) Tissue unfixed 2) tissue fixed, then processed with by whatever step was supposed to neutralize the fixation effect, rinsing the tissues carefully afterwards. . Then I would run tissues 1 and 2 in the same preparation. And I would choose as my standard procedure which ever procedure of the pair which gave the better results. I drop that and run. The crowding effect of Standard Procedure Itis is out there. But then I might just find my question was trivial and standard procedure might win. But has the fix-unfix procedure ever been tested against the unfixed prep? This retired picky-compulsive tech would never run ANY procedure as standard without testing its effectiveness---comparing it--- with any reasonable alternative procedure for quality. Nuff said. Retired Tech now enters a stage of quiet upon the subject on the histonet, but I will still brood over the untapped quality test in ANY procedure done by rote rather than via the search for the most excellent way to do that test. georgecole@ev1.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joeamateur <@t> hotmail.com Thu Sep 16 14:28:15 2004 From: joeamateur <@t> hotmail.com (Jack England) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] vascular graft_sectioning difficulty Message-ID: Hmm. If the tissue is coming out well, then processing and embedding seem fine; it sounds like the Dacron itself is brittle during cutting. Are you observing the Dacron graft to be more brittle after processing than before? I agree with Melanie about chilling the paraffin blocks or resin embedding. If you're not already doing so, maybe try cutting with a high profile blade, and/or a shallower blade angle? --Aloha, Jack England >From: "Dr. Raghul" >To: joeamateur@hotmail.com,Histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] vascular graft_sectioning difficulty >Date: Thu, 16 Sep 2004 17:02:16 +0530 > >Hello Aloha, > > Our vascular graft material(DACRON) after fixing in 10% NBF, we >dehydrate in ascending grades of alcohol(70,80,95,100%),cleared in 3 >changes of chloroform before embedding in paraffin wax (60*C MPT). > > While cutting, the graft area is powdering in the sections >obtained.When it comes to the anastomising graft-host vessel part, while >the vessel portion comes well, the graft as usual gets powdered . > > This is our processing detail and the sectioning difficulty we >face.Still if the details are insufficient please ask. > >regards >raghul > >-----Original Message----- >From: "Jack England" >To: raghul@sctimst.ker.nic.in, Histonet@lists.utsouthwestern.edu >Date: Wed, 15 Sep 2004 08:54:40 -1000 >Subject: RE: [Histonet] vascular graft_sectioning difficulty > > > Can you provide a little more information about your processing, and a > > little more detail about how the sections are crumbling (eg, tissue is > > disaggregating, or the paraffin section is falling apart, etc)? We > > regularly > > section formalin-fixed, paraffin-embedded vascular grafts (ePTFE) at > > 5-7µm, > > and it's working for us. With a little more information, I might be > > able to > > help... > > > > Aloha, > > Jack England > > Tissue Genesis, Inc. > > http://www.tissuegenesis.com > > > > >From: "Dr. Raghul" > > >To: histonet@lists.utsouthwestern.edu > > >Subject: [Histonet] vascular graft_sectioning difficulty > > >Date: Tue, 14 Sep 2004 16:56:23 +0530 > > > > > >hello members, > > > > > >We cut 5 µm thin sections of paraffin embedded vascular graft(Dacron) > > in > > >a rotary microtome with disposable blades but the senior tech tells > > that > > >the tissue sections are crumbling and forces us to go for thick > > sections > > >around 10µm but again not with much success. > > > > > > We can understand that there is not enough tissue adhereing to the > > >graft and we are literally cutting the graft. But did anyone had such > > >problems sectioning vascular grafts. Any suggestions? > > > > > > > > >regards , > > > > > >Raghul J > > >scientist > > >Histopathology- implant biology division > > >Srichitra tirunal institute of medical sciences and technology > > >Trivandrum -695 012 > > >Kerala, India > > > > > > > > >_______________________________________________ > > >Histonet mailing list > > >Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _________________________________________________________________ > > FREE pop-up blocking with the new MSN Toolbar – get it now! > > http://toolbar.msn.click-url.com/go/onm00200415ave/direct/01/ > _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From TJasper <@t> smdc.org Thu Sep 16 18:04:55 2004 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:24:03 2005 Subject: FW: [Histonet] Position Opportunity Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E44FB3@harrier> I am reposting this as we are still looking for that "special someone" that I know is out there in Histonetland. I will admit to being biased to living in this beautiful part of the world, but we do have a great opportunity available here. Our group is very dedicated, fun-loving and nice to work with. I will also give high marks to our pathologists. Not only are they top-notch diagnostically, they are cognizant of the fact that the technical staff is skilled and intelligent. We work very closely with them and they are very appreciative of our efforts day in and day out. Thanks for taking the time to consider joining our service. Most sincerely, Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Jasper, Thomas G. [mailto:TJasper@smdc.org] Sent: Wednesday, August 11, 2004 5:19 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] Position Opportunity Opportunity available in Duluth, MN. SMDC (St. Mary's/Duluth Clinic) seeks to fill a position in the Histology Department. This is a full-time day shift position at a progressive tertiary care facility. SMDC is located near the shore of scenic Lake Superior. Duluth is a clean and green community within minutes of wilderness in both northern Minnesota and Wisconsin. Greater urban amenities are available in Minneapolis/St. Paul 150 miles south on I-35. Duluth offers much in the way of culture, education and entertainment in the greater western Lake Superior area. SMDC serves northern Minnesota, northern Wisconsin and the western Upper Peninsula of Michigan. Consider joining our respected and respectful pathology service, here at our hidden gem. For further information and application information log on to www.smdc.org . Phone 218/786-4564 Fax 218/786-4018. Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Gervaip <@t> aol.com Thu Sep 16 20:02:19 2004 From: Gervaip <@t> aol.com (Gervaip@aol.com) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] unsubscribing Message-ID: From jnocito <@t> satx.rr.com Thu Sep 16 21:35:03 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] endogenous peroxidase blocking protocol References: <20040916145628.72162.qmail@web52609.mail.yahoo.com> Message-ID: <017b01c49c5e$f0396a50$4661ce44@yourxhtr8hvc4p> Ann, have you tried a 3% in methanol solution? Depending on the number of slides you have. I used to use 450 mL methanol to 50mLs H2O2 on really bloody specimens. You might have to perform a time titer like 5 min., 10 min, 15 min. Good luck Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Ana Florentin" To: Sent: Thursday, September 16, 2004 9:56 AM Subject: [Histonet] endogenous peroxidase blocking protocol > Hi, > Does anybody know a good protocol for blocking the > endogenous peroxidase? > I am performing immunoperoxidase staining on rat > spleen frozen sections and I have tried to block > endogenous peroxidase with H2O2, 1% and 0.3% in PBS, > half hour before the first antibody but is seemed to > be a too high concentration, as the tissue was > destroyed. > Thanks for help! > anne > > > > > __________________________________ > Do you Yahoo!? > New and Improved Yahoo! Mail - 100MB free storage! > http://promotions.yahoo.com/new_mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From bharatm <@t> glenmarkpharma.com Thu Sep 16 22:52:30 2004 From: bharatm <@t> glenmarkpharma.com (Bharat Mani) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Eosinophils in mouse tissue Message-ID: <2E1FBE2AC0130748B591FD48AC7056E36221FE@e2k1mhp.glenamrk.com> Dear Todd We do a lot of experiments on eosinophilia in mouse, ensure to perfuse the lungs before processing, am sharing our protocol - 1. We use 10% NBF to perfuse the mouse lungs and then subsequently proceed for immersion fixation for atleast a minimum of 24-48 hours before processing the mouse lung/s tissue. 2. Next we do a routine H&E on sections cut at 5?. Hope this helps Regards Bharat Mani Research Scientist (Biological Research) Glenmark Research Centre Plot No. A/607, TTC Ind. Area, MIDC, Mahape Navi Mumbai - 400 709 Ph.: +92-22-5590 2491/92 Ext.: 235 Fax: +91-22-5590 2318 Email: bharatm@glenmarkpharma.com Web: www.glenmarkpharma.com -----Original Message----- From: Histology SLU [mailto:sluhisto@yahoo.com] Sent: Friday, September 17, 2004 1:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Eosinophils in mouse tissue Does anyone out there have a protocol for staining eosinophils in mouse lung? Thanks. Todd --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: The information contained in this email communication is intended only for the personal and confidential use of the designated recipient named above. This message may be an attorney-client communication, and as such is privileged and confidential. If the reader of this message is not the intended recipient, you are hereby notified that you have received this communication in error, and that any review, dissemination, distribution, or copying of the message is strictly prohibited. If you have received this transmission in error, please destroy this transmission and notify Glenmark Pharmaceuticals immediately by telephone and/or send an email to supportho@glenmarkpharma.com From ian.montgomery <@t> bio.gla.ac.uk Fri Sep 17 03:58:29 2004 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Eosinophils. Message-ID: <6.1.0.6.2.20040917095139.02d3c790@udcf.gla.ac.uk> Todd, The most consistent method for mouse tissue, in my hands, is the Sirius Red technique developed by Brian Llewellyn. Llewelyn,B.D. (1970) J. Med. Technol. 27. 308-309. An improved sirius red method for amyloid. If you cannot get the original paper e-mail and I'll give you the technique. 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 jluis.palazon <@t> icman.csic.es Fri Sep 17 05:02:37 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Gomori chrome alum hematoxylin Message-ID: <20040917100237.2DAAE2CF1C2@perceval.uca.es> Good day dear L-members Many thanks for all the fellow members who answered my question about chrome alum hematoxilin regards Jos? Luis El dia 15/09/2004 20:20 usted envio el siguiente mensaje: >Date: 15 de Septiembre de 2004 20:20:22 >From: Bryan Llewellyn >Subject: Re: [Histonet] Gomori chrome alum hematoxylin >To: histonet@lists.utsouthwestern.edu > > According to Gray it is:- > > Water - 100 mL > Sulphuric acid, 5% aqu. - 2 mL > Potassium dichromate 5% aqu. - 2 mL > Chrome alum - 1.5 g > Hematoxylin - 0.5 g > > Dissolve the alum and the hematoxylin each in 50 mL water. > Combine the two solutions. > Add the potassium dichromate and sulphuric acid. > Leave for 48 hours to ripen. > > Bryan Llewellyn > > > Jose Luis Palazon Fernandez wrote: > > Dear list-members > > > > > > > > Greetings. I can not find the recipe to prepare Gomori?s chrome alum hematoxylin. Could any of you help me please? > > > > > > > > Thanks in advance > > > > > > > > > > > > Jos? Luis > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > 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 gu.lang <@t> gmx.at Fri Sep 17 05:43:16 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] movat question Message-ID: <002901c49ca3$29a9c250$eeeea8c0@server> Hi! I'm going to perfom the pentachrom movat stain for the first time. We will use it with lung-tissue. Can anyone give me usefull hints and tricks? Any easy procedure is also appreciated. thank you Gudrun From ktalbot <@t> jhmi.edu Fri Sep 17 07:33:43 2004 From: ktalbot <@t> jhmi.edu (Karen Fox-Talbot) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] CD31/PECAM in Rat Brain Histonet Digest Message-ID: re: CD31/PECAM in Rat Brain Hi Danielle, I use a polyclonal goat anti mouse CD 31 from Santa Cruz (SC 1506) to stain for Pecam in Rat tissue. this antibody cross reacts with human, mouse and rat . I have also used it in pig. My tissues are fixed in 60% methanol/10% glacial acetic acid. karen Karen Fox-Talbot MAT The Johns Hopkins University School of Medicine Department of Pathology Immunohistochemistry Lab 410.614.0915 Lab 410.283.9307 Pager From Sue.Kapoor <@t> uhsi.org Fri Sep 17 09:30:48 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Energy Beam microwave processor Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F5664@khmcexch.uhsi.org> Hi, does anyone in histoland use an Energy Beam microwave tissue processor? I'm doing a demo of one now and have some questions. Thanks, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From ploykasek <@t> phenopath.com Fri Sep 17 09:54:46 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Mayers problem Message-ID: Hi all. Of course when we're extremely short-handed we have a problem with the hemo. Thought I'd ask for help before my brain goes kaput. We make our own Mayer's hemo & yesterday it was muddy & too purple. We tried a couple of batches - all with same results. The 1 new lot that we have is the aluminum ammonium sulfate. I think that is the culprit. Does this make sense to anyone else? I'm open to other ideas, too. Patti Loykasek PhenoPath Laboratories Seattle, WA From chris <@t> ptplab.com Fri Sep 17 10:19:19 2004 From: chris <@t> ptplab.com (Christopher L. Robertson) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Histology lab lists Message-ID: Hello everyone. Does anyone know where I can get a complete list of histology laboratories in the United States? I can't seem to find anything on the web. Thanks in advance. Chris PTPLAB Portland, Oregon From daniel.eberhard <@t> uni-bielefeld.de Fri Sep 17 11:13:02 2004 From: daniel.eberhard <@t> uni-bielefeld.de (DANIEL EBERHARD) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] anti Desmin AB Message-ID: <414B0D0E.3080200@UNI-BIELEFELD.DE> Dear All, what?s your favorite anti-Desmin AB for pFA-fixed mouse sections? Thanks Daniel From sluhisto <@t> yahoo.com Fri Sep 17 11:58:20 2004 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Kappa Lambda Message-ID: <20040917165820.83945.qmail@web51005.mail.yahoo.com> Hello all: What we have is Lambda that works on B5 fixed clot and core sections and Kappa that only works on the B5 fixed clots. We are getting nada in the B5 decaled cores. We use B5 fixed tonsil as control and we put our patient samples on the same slide as the control. Our controls are all working appropriately. Any thoughts????? We use Dako Envision + system and are using protease K for 5 minutes(any longer and our clots are eaten off). The only variable that we have is that the cores are of course decaled for one hour. All comments and suggestions are welcome. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From TJasper <@t> smdc.org Fri Sep 17 12:01:28 2004 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] fixed and unfixed immuno and antuibody oreps Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E44FB4@harrier> George, I am sorry if I seem naive but I do understand your point about unfixed in the first place, then not having to antigen retrieve to compensate for a new problem which has been created. Perhaps this is more a difference in application to achieve a goal. It's certainly possible in a purely academic setting that the unfixed approach could produce the best and most direct results. What has happened in clinical settings is that IHC has been developed for use after tissue has been fixed. The fixation occurs out of necessity, not for the IHC, but for the routine Hematoxylin and Eosin work-ups. Many times what pathologists see on H+E staining determines if IHC is necessary to diagnose a case. At gross examination, before the histological work-up, the pathologist may already want to order some antibodies prior to seeing the H+E. Nevertheless, fixation is necessary as it is a required step in the routine work-up for tissue specimens in a case. Keep in mind that time and cost effectiveness are involved. If a case can be diagnosed on H+E only or on H+E and special stains (non-IHC) it will be done. Most certainly pathologists were diagnosing cases on H+Es and limited special stains long before the development of IHC. Routine fixation for histological work-up is basically a clinical standard of care. There is another practical consideration with unfixed tissue. To my understanding that would require freezing everything that you were interested in, or potentially interested in for IHC. Freezing tissue presents a set of unique problems on it's own. There are things such as freezing artifact and tissues that just do not freeze well (anything fatty). There are also specimens that cannot be practically frozen due to calcification. Clinically, even with exceptionally good frozen technique, most pathologists in many instances will defer to "permanents" once the routine work-ups are done. In many instances surgeons will get exclusionary information or a limited list of possible "what-ifs" from a pathologist until further information is available. George, I would never claim that what we do in the clinical world is the best or most perfect way to demonstrate antigenicity in tissue specimens. What I will say is for the most part this is what we do and why is because this is the best way that we know how. Again, I by no means am expert in the field of Immunohistochemistry and please allow me the disclaimer to be corrected about anything that I have posted. As I've said before there are many out in Histonetland that are have more knowledge than I do. I guess I'm just trying to help you understand what goes on from a standard clinical perspective with IHC. Regards, Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: George Cole [mailto:georgecole@ev1.net] Sent: Thursday, September 16, 2004 9:47 PM To: 'Jasper, Thomas G.' Subject: RE: [Histonet] fixed and unfixed immuno and antuibody oreps Thomas; The note you answer so thoughfully is na?ve, I admit---I mean it says HMMM, if they are undoing the effects of fixatives on their tissues----wouldn't NOT fixing in the first place them be an even better way to undo fixation? That's what it is to be retired with no lab to fiddle with questions like that. There is only the Histonet---may be I can get somebody out there to wrestle with this question----sic 'em on it, so to speak---maybe somebody will soothe this tech, retired, yes, but still with questions buzzing around his aged head. georgecole@ev1.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jasper, Thomas G. Sent: Thursday, September 16, 2004 10:45 AM To: 'George Cole'; 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] fixed and unfixed immuno and antuibody oreps Dear George, I have read some of your postings and I have developed a soft spot for you. It seems what you're after is an explanation of how immunohistochemistry can be done these days using techniques which negated your efforts in the past. Also you seem to be questioning how the "best" method was determined, why is it OK to do IHC the way most labs do? George, I honestly cannot answer how today's techniques were exactly determined/developed. I am not an expert in the field of IHC history and comparative research of the same. Here's what I do know, I have been doing Histology work for approximately 20 years (I know a babe in woods). All of my training and experience in doing IHC has been heavily dependent upon proper fixation. This is for the most part formalin fixed (10% Neutral Buffered), paraffin processed tissue specimens. Most people working in the clinical world, as I do, base the majority of their IHC work on properly fixed tissue specimens. This is how we understand IHC. My lab runs into problems occasionally when we receive referral cases (either block or slides) that have poor/improper fixation. This hinders our ability to produce a good quality diagnostic stain. Antigen retrieval techniques are employed to enhance staining and maybe that is the answer you seek as it is a post-fixation procedure. George, I am not a researcher and there are a lot of people on the histonet that are better qualified than I am to speak to the finer points of IHC. I just thought I would share my thoughts with you. I consider myself to be a standard histologist with a fairly basic understanding of IHC as it relates to I what I am expected to put out on a daily basis. George, I apologize if the things I have just stated have already been related to you or if someone with a greater understanding of IHC finds flaws in what I've posted. Being a manager I don't do quite as much IHC as I used to, although I certainly have my hand in the day-to-day immunos that are run here. I also sympathize with you in a general sense as I do like to understand things and like clear explanations and/or reasons why. Hope this helps. Regards, Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: George Cole [mailto:georgecole@ev1.net] Sent: Tuesday, September 14, 2004 1:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fixed and unfixed immuno and antuibody oreps I promise to drop this line---I have had an itch to find out something--- I realize I must give it up---I have asked it repeatedly and I don't want to keep asking the same thing---impatience is bound to develop. Retired as I am---I no longer have lab in which to answer such questions. Most histotechs nowadays are brought up doing one side of the question---indeed it is the standard procedure and my question just doesn't scan with today's techs. But if I could shuck 10 years and be back in my lab and I was given the responsibility to do a certain class of work I would do the following: Antibody technique or immunofluorescence procedure---any procedure that has a fixative in its beginning followed by some procedure to neutralize the effects of the fixative----I would run at least 3 trials on the 2 sets of pairs: 1) Tissue unfixed 2) tissue fixed, then processed with by whatever step was supposed to neutralize the fixation effect, rinsing the tissues carefully afterwards. . Then I would run tissues 1 and 2 in the same preparation. And I would choose as my standard procedure which ever procedure of the pair which gave the better results. I drop that and run. The crowding effect of Standard Procedure Itis is out there. But then I might just find my question was trivial and standard procedure might win. But has the fix-unfix procedure ever been tested against the unfixed prep? This retired picky-compulsive tech would never run ANY procedure as standard without testing its effectiveness---comparing it--- with any reasonable alternative procedure for quality. Nuff said. Retired Tech now enters a stage of quiet upon the subject on the histonet, but I will still brood over the untapped quality test in ANY procedure done by rote rather than via the search for the most excellent way to do that test. georgecole@ev1.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Sep 17 12:52:37 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] CPT Code Help Message-ID: <83AACDB0810528418AA106F9AE9B7F7E2782FC@sjhaexc02.sjha.org> I think I've completely lost my mind! I believe that I recently read something regarding bundling and unbundling specimens for CPT coding that states that a thyroid gland, even if it is received in two containers, should be billed with one specimen only. Yet, I am unable to to recall where I read it. I have searched the archieves and don't find anything there. Could someone shed some light on this please? One total thyroid - one CPT code? Thanks in advance. Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From JGordon <@t> cellmarque.com Fri Sep 17 14:19:42 2004 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] P-63 help please!! (FREE p63 offer) Message-ID: Jesus, I'm sorry for the original inconvenience, but I hope that this helps. Due to the recent unavailability of p63 for purchase, Cell Marque, in its commitment to serve its clients in the best manner possible, is offering the anti-p63 antibody at no additional charge for a limited time. This antibody will be sent with any outgoing orders that include a request for the anti-p63. The antibody will be sent out in a 1 ml prediluted vial, good for up to 10 tests. There is a limit of one FREE anti-p63 vial that can be included with each purchase order; however, while this offer lasts, there is no limit to the amount of free anti-p63 antibody that a lab can receive in any time period. To take advantage of this limited time offer, just include catalog number CMA441 on future Cell Marque orders and you will receive the anti-p63 at no additional charge. Jeff Gordon Cell Marque Corp. 1-800-665-7284, Ext. 12 http://www.cellmarque.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jesus Ellin Sent: Tuesday, September 07, 2004 9:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] P-63 help please!! Currently we are in a dilema we are looking for a P-63 antibody that works well with thet Ventanna stainers. We use to buy it fomr cell marque and they are now discontinuing thier product. If any one can help with this it would be greatly appreciated. The clone that we are currently using is a mouse monoclonal clone (4A4). Thanks Jesus Ellin Yuma Regional Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From joycejudge259 <@t> hotmail.com Fri Sep 17 14:25:51 2004 From: joycejudge259 <@t> hotmail.com (joyce judge) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] sections on scotch tape Message-ID: I may have sent this twice but anyway. I am looking for info on immuno staining sections on scotch tape. I know Instrumedics has a great tape transfer system but we are unable to purchase it at this time. I have nice sections of undecalcified mouse paws on scotch tape and I am looking for help doing immunos on this tape. Any help would be greatly appreciated. I have checked the archives. Thanks, Joyce Judge Visen Medical _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From lkbauer <@t> unmc.edu Fri Sep 17 15:05:50 2004 From: lkbauer <@t> unmc.edu (lkbauer@unmc.edu) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] DAF-FM DA Message-ID: I would like to visualize Nitric Oxide production in paraffin sections of mouse embryos. I was thinking of using DAF-FM DA from Molecular Probes. Has anyone had any luck(good)with this reagent? Thanks, Lin Linda(Lin) Bauer 985455 Nebraska Medical Center Omaha, NE 68198-5455 phone: (402) 559-2863 fax: (402) 559-4001 email: lkbauer@unmc.edu From naje1972 <@t> yahoo.com Fri Sep 17 16:30:12 2004 From: naje1972 <@t> yahoo.com (cynthia haynes) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Service Manual/Shandon-Hypercente Message-ID: <20040917213012.68814.qmail@web41711.mail.yahoo.com> Hello everyone, Is there anyone out in Histoland that has a service manual for the Shandon-HypercenterXP. We would be happy to purchase it from you if possible. Please contact me at 773-342-1559 and please leave a message and I will definitely get back to you. Thanks in advance. Cynthia Haynes H.T. Laboratory Directory __________________________________ Do you Yahoo!? Yahoo! Mail - You care about security. So do we. http://promotions.yahoo.com/new_mail From Linresearch <@t> aol.com Fri Sep 17 18:06:33 2004 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Phos. Antibodies Message-ID: <92.15751f8c.2e7cc7f9@aol.com> Greetings, is there anyone in Histoland who has knowledge of protocols for Phosphorylated Abs on FFPE rodent tissues. Could you share that information? lin From histophilhuff <@t> yahoo.com Fri Sep 17 20:10:45 2004 From: histophilhuff <@t> yahoo.com (Phillip Huff) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Disposable Blades for Cutting Frozen Skin - Recommendations Message-ID: <20040918011045.9090.qmail@web50301.mail.yahoo.com> What disposable blades have people had success using for the cutting of skin samples? I would like to have people's suggestions for the best disposable cryostat blade for skin or other such tissue. Thanks! Phil __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From portera203 <@t> yahoo.com Fri Sep 17 20:47:06 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] endogenous peroxidase blocking protocol In-Reply-To: <20040916145628.72162.qmail@web52609.mail.yahoo.com> Message-ID: <20040918014706.78629.qmail@web40908.mail.yahoo.com> Anne - maybe try 0.03% in methanol. It works pretty well for me with post fixed frozen sections of human tonsil. Ana Florentin wrote:Hi, Does anybody know a good protocol for blocking the endogenous peroxidase? I am performing immunoperoxidase staining on rat spleen frozen sections and I have tried to block endogenous peroxidase with H2O2, 1% and 0.3% in PBS, half hour before the first antibody but is seemed to be a too high concentration, as the tissue was destroyed. Thanks for help! anne __________________________________ Do you Yahoo!? New and Improved Yahoo! Mail - 100MB free storage! http://promotions.yahoo.com/new_mail _______________________________________________ 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!? vote.yahoo.com - Register online to vote today! From tahseen <@t> brain.net.pk Fri Sep 17 21:31:07 2004 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Antibody for Langerhans' Cell Histiocytosis Message-ID: <005b01c49d27$8e86c9a0$972bfea9@m7c0y4> Hello all, Does anyone know of an antibody for Langerhans' Cell Histiocytosis on paraffin embedded samples other then CD1a (MAb O10). Thank you all for your time Muhammad Tahseen. From joycejudge259 <@t> hotmail.com Fri Sep 17 14:21:23 2004 From: joycejudge259 <@t> hotmail.com (joyce judge) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] IHC bone sections on scotch tape Message-ID: Does anyone have any experience staining sections on scotch tape? I know Instrumedics has a tape transfer system that works well but we are unable to purchase it at this time. I have nice sections of undecalcified mouse paws on scotch tape and need to do some immuno. Has anyone tried this? I have checked the archives and I don't find anything. Any help would be appreciated. Joyce Judge Visen Medical _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From wrhewett <@t> austarnet.com.au Sat Sep 18 02:29:24 2004 From: wrhewett <@t> austarnet.com.au (william hewett) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] kappa/lambda Message-ID: <015d01c49d51$3a2b13f0$7fcbdccb@hewett> Susan We do kappa/lambda on trephine marrows using Dako envision and proteinase -k also.The predilute works well for both, diluted 1/2 using reactive lymph node as control. Bill Hewett QHPS Laboratories Townsville Australia. From wrhewett <@t> austarnet.com.au Sat Sep 18 02:47:18 2004 From: wrhewett <@t> austarnet.com.au (william hewett) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] microwriters Message-ID: <019401c49d53$ba0395a0$7fcbdccb@hewett> Hi histonetters, Is anyone using , or has experience with carousel cassette microwriters.?These machines permanently mark tissue cassettes using foil tape and a heated stylus with identifying numbers.Different coloured cassettes can be mounted in each vertical hopper to code various types of specimens at cut-up. Bill QHPS Lab. T'ville. Aust. From JColCLEFA <@t> aol.com Sat Sep 18 10:37:17 2004 From: JColCLEFA <@t> aol.com (JColCLEFA@aol.com) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Storage,double stains Message-ID: <82.16605026.2e7db02d@aol.com> We store blocks and slides offsite once they are over 10 years old. To consolidate material, after 10 years we discard blocks from negative cases, but always save the slides. We do doublestains on the Biogenex i6000 using the research software key, and before we wrestled the key from the vendor, by keeping a map of "nominal steps" vs "real steps" on a chart next to the instruments. They work well that way (the stains that is) Our lab does Pathvysion Her 2-- ask away, private e-mail ok From gu.lang <@t> gmx.at Sat Sep 18 11:08:22 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] movat question Message-ID: <000a01c49d99$b8d8a760$eeeea8c0@server> > Hi Jaqueline, > I have looked through all literature I could get. Can you tell me what's the > right dye: Crocein Scarlet (CI 27165) or Biebrich Scarlet (CI 26906) or > Brilliant Crocein Scarlet MOO (CI 27290). > It is not so easy to find the right one in the right German translation. > Perhaps you can help me? > tsch?s > Gudrun > > > > ----- Original Message ----- > From: "Jacqueline Cruz" > To: "Gudrun Lang" > Sent: Friday, September 17, 2004 3:58 PM > Subject: Re: [Histonet] movat question > > > > Hi Gudrun, > > I love that stain! It's beautiful, and easy to perform. I will admit it > is > > long (about 4 hrs, if memory serves). No short cuts that I know of. Good > > luck. > > Jackie > > ----- Original Message ----- > > From: Gudrun Lang > > To: Histonetliste > > Sent: Friday, September 17, 2004 6:43 AM > > Subject: [Histonet] movat question > > > > > > Hi! > > I'm going to perfom the pentachrom movat stain for the first time. We will > > use it with lung-tissue. > > Can anyone give me usefull hints and tricks? Any easy procedure is also > > appreciated. > > > > thank you > > Gudrun > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > From carl.hobbs <@t> kcl.ac.uk Sat Sep 18 13:12:52 2004 From: carl.hobbs <@t> kcl.ac.uk (carl hobbs) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] re anti Desmin AB Message-ID: Dako M 724 and M760 with AR .beautiful on mouse, rat and chicken! Be interesting to hear of any caveats . --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.762 / Virus Database: 510 - Release Date: 13/09/2004 From carl.hobbs <@t> kcl.ac.uk Sat Sep 18 13:36:59 2004 From: carl.hobbs <@t> kcl.ac.uk (carl hobbs) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] endogenous peroxidase blocking protocol Message-ID: Agree with Amy. NO aq solns for frozen section endog px block. Alcoholic or, if you are concerned re alcohol effects on antigenicity, use glucose oxidase methos, as recommended by Gayle in a previous post. Or alk. Phos detection system ( with levamisol block, if pertinent) --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.762 / Virus Database: 510 - Release Date: 13/09/2004 From mikael.niku <@t> helsinki.fi Mon Sep 20 01:16:21 2004 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Digital camera setup for histology teaching In-Reply-To: Message-ID: <000a01c49ed9$59a4c940$8c0fd680@ekk1116> Dear Histonetters, we are planning to replace our old histology teaching equipment (an analog video camera attached to the teacher's microscope + monitors for the students) with new digital stuff (digital video camera + data projector). Please recommend a setup. I have the following requirements in my mind: - The idea is to display the view from the teacher's microscope on the screen. - We need LIVE image of course. - A computer software supporting the teaching would be nice (for example, allowing the teacher to draw on the live image, and perhaps showing several simultaneous images from the memory on the screen). - Max resolution of the projector is 1280x1024, so the camera doesn't need a superb resolution. Good color reproduction is necessary, though, as well as good automatic brightness adjustment. - We need to fit the camera on the old Olympus BH2. Thanks in advance. //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// From abright <@t> brightinstruments.com Mon Sep 20 02:49:20 2004 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] sections on scotch tape Message-ID: I too would be very interested. 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: joyce judge [mailto:joycejudge259@hotmail.com] Sent: 17 September 2004 20:26 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] sections on scotch tape I may have sent this twice but anyway. I am looking for info on immuno staining sections on scotch tape. I know Instrumedics has a great tape transfer system but we are unable to purchase it at this time. I have nice sections of undecalcified mouse paws on scotch tape and I am looking for help doing immunos on this tape. Any help would be greatly appreciated. I have checked the archives. Thanks, Joyce Judge Visen Medical _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From barbara.bublava <@t> meduniwien.ac.at Mon Sep 20 03:48:15 2004 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Glycerin-Gelatine bubble-phenomen Message-ID: <01cd01c49eee$91bc2c50$1401a8c0@GERICHTS9XOZZ8> Hi Everybody! I have an interesting phenomen mounting slides with Glyceringelatine When I mount my slides they do not have bubbles. Oil Red O stains do not develop bubbles even after some time but when I mount CAE - Chloracetate Esterase (I mount in the same way I do the ORO) the slides develop bubbles after some hours and after some days the whole section is covered with bubbles, but there are no bubbles around the section! Why does this happen and how can I prevent this??? I use Glyceringelatine at 60?C, slides out of dest. water, excess water shaked of. I did also try to dry the slides bevore mounting but there happend to be more bubbles from the beginning and it did not change this strange effect. Sometimes, when I am in a hurry I further hardening of the Glyceringelatine at fridge (4?C) but I can not see any positive or negative effect on bubbles doing that. Thanks to everyone Barbara, Vienna From Lawrence.Brett <@t> luht.scot.nhs.uk Mon Sep 20 05:06:17 2004 From: Lawrence.Brett <@t> luht.scot.nhs.uk (Brett, Lawrence) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] p63 Message-ID: <857344E7D8F90240935A288998A89D811C6045@wgh-ex1.luht.scot.nhs.uk> We get our p63 from Dako in the UK. Code no. M7247, clone 4A4. Works well on paraffins with heat retrieval. Lawrence Brett Edinburgh Scotland UK ********************************************************************** The information contained in this message may be confidential or legally privileged and is intended for the addressee only, If you have received this message in error or there are any problems please notify the originator immediately. The unauthorised use, disclosure, copying or alteration of this message is strictly forbidden. ********************************************************************** From Barry.R.Rittman <@t> uth.tmc.edu Mon Sep 20 06:38:23 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Glycerin-Gelatine bubble-phenomen Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0F35265@UTHEVS3.mail.uthouston.edu> I believe that part of the problem is that you are drying the slides before adding the glycerin gelatin. I would recommend shaking excess water from the slide, placing the slide on a warming plare at 60 degrees C. Add a few drops of glycerin gelatin and leave for a few minutes to allow any bubbles to rise to the surface. Then add coverslip. The bubbles are probably coming from the glycerin as sections mounted just in glycerin always develop bubbles after some days. Suggestion is to place the glycerin in a tube in an ultrasonic bath to get bubbles out before making the glycerin gelatin or simply doing the same to the molten glycerin gelatin. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Barbara Bublava Sent: Mon 9/20/2004 3:48 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Glycerin-Gelatine bubble-phenomen Hi Everybody! I have an interesting phenomen mounting slides with Glyceringelatine When I mount my slides they do not have bubbles. Oil Red O stains do not develop bubbles even after some time but when I mount CAE - Chloracetate Esterase (I mount in the same way I do the ORO) the slides develop bubbles after some hours and after some days the whole section is covered with bubbles, but there are no bubbles around the section! Why does this happen and how can I prevent this??? I use Glyceringelatine at 60?C, slides out of dest. water, excess water shaked of. I did also try to dry the slides bevore mounting but there happend to be more bubbles from the beginning and it did not change this strange effect. Sometimes, when I am in a hurry I further hardening of the Glyceringelatine at fridge (4?C) but I can not see any positive or negative effect on bubbles doing that. Thanks to everyone Barbara, Vienna _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Mon Sep 20 07:08:25 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Disposable Blades for Cutting Frozen Skin -Recommendations Message-ID: Accuedge low profile. Hard to beat. I once did a project where I had to cut over 500 frozen skin bx's without skipping a beat. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 -----Original Message----- From: Phillip Huff [mailto:histophilhuff@yahoo.com] Sent: Friday, September 17, 2004 8:11 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Disposable Blades for Cutting Frozen Skin -Recommendations What disposable blades have people had success using for the cutting of skin samples? I would like to have people's suggestions for the best disposable cryostat blade for skin or other such tissue. Thanks! Phil __________________________________________________ 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 Barry.R.Rittman <@t> uth.tmc.edu Mon Sep 20 08:03:32 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Antibody for Langerhans' Cell Histiocytosis Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0F135C7@UTHEVS3.mail.uthouston.edu> Muhammad I have never used the Langerin antibody for demonstration of Langerhans cells but have been told that it is reliable. Two references only one for confocal microscopy and other undetermined. May be a string point for you. Lamarque S. Pellen-Mussi P. Rougier N. Le Lan J. Chesne C. Bonnaure-Mallet M. Gingival organotypic culture and langerhans cells: a tool for immunotoxicologic experiments. Journal of Biomedical Materials Research. 68A(2):257-63, 2004 Feb 1. Allam JP. Geiger E. Kraft S. Bieber T. Limited reliability of E-cadherin as a specific marker for in vitro-generated Langerhans cells. Archives of Dermatological Research. 295(6):263-8, 2003 Nov. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Muhammad Tahseen Sent: Friday, September 17, 2004 9:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Antibody for Langerhans' Cell Histiocytosis Hello all, Does anyone know of an antibody for Langerhans' Cell Histiocytosis on paraffin embedded samples other then CD1a (MAb O10). Thank you all for your time Muhammad Tahseen. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From stevemachinuk <@t> yahoo.co.uk Mon Sep 20 08:13:51 2004 From: stevemachinuk <@t> yahoo.co.uk (Steve Machin UK) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Digital camera setup for histology teaching In-Reply-To: <000a01c49ed9$59a4c940$8c0fd680@ekk1116> Message-ID: <20040920131351.34303.qmail@web25104.mail.ukl.yahoo.com> We use the video output from a nikon coolpix. The video goes to a Leitz unit with a joystick on it, to act as a pointer, and this box has an output for a TV screen and a splitter for a couple of video monitors. Our previous system used an analogue camera and we simply substituted this by the coolpix. Steve machin UK --- Mikael Niku wrote: > Dear Histonetters, > > we are planning to replace our old histology teaching equipment (an > analog video camera attached to the teacher's microscope + monitors > for > the students) with new digital stuff (digital video camera + data > projector). > > Please recommend a setup. > I have the following requirements in my mind: > > - The idea is to display the view from the teacher's microscope on > the > screen. > > - We need LIVE image of course. > > - A computer software supporting the teaching would be nice (for > example, allowing the teacher to draw on the live image, and > perhaps > showing several simultaneous images from the memory on the screen). > > - Max resolution of the projector is 1280x1024, so the camera > doesn't > need a superb resolution. Good color reproduction is necessary, > though, > as well as good automatic brightness adjustment. > > - We need to fit the camera on the old Olympus BH2. > > Thanks in advance. > > > //////////////////////////////////////////////////////////// > > Mikael Niku URL: www.helsinki.fi/~mniku/ > University of Helsinki Dept. Basic Veterinary Sciences > > - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? > Minusta se olisi erinomainen ajatus! > > - Gandhi > > //////////////////////////////////////////////////////////// > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ___________________________________________________________ALL-NEW Yahoo! Messenger - all new features - even more fun! http://uk.messenger.yahoo.com From jtsonger <@t> vt.edu Mon Sep 20 09:34:52 2004 From: jtsonger <@t> vt.edu (Jill Songer) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] yersinia pestis Message-ID: <6.0.0.22.0.20040920103256.01b375a0@pop.vt.edu> Does anyone out there in cyber land know the best stain for this bacteria? Thanks in advance. 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 MICHAEL.OWEN <@t> fda.gov Mon Sep 20 10:07:39 2004 From: MICHAEL.OWEN <@t> fda.gov (Owen, Michael P) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Yersinia pestis Staining Message-ID: Dear Jill, According to the Sentinel Procedures (previously called Level A) of the CDC Laboratory Response Network (LRN), Gram and Wright-Giemsa Stains are performed on specimens to make a presumptive identification of Yersinia pestis. Consult the Web pages below for details. CDC Emergency Preparedness and Response: Plague Information http://www.bt.cdc.gov http://www.bt.cdc.gov/agent/plague/index.asp The Reference Procedures (previously called Levels B and C) of the LRN are restricted to member organizations in the LRN. They are considered sensitive material and are not available for public distribution. The LRN Reference Procedures describe other staining tests such as direct fluorescent antibody staining. Historically, the Wayson Stain has been used during the identification process of Yersinia pestis. The Web page below has a nice photograph of the stain. CDC Division of Vector-Borne Infectious Diseases: Wayson Stain Image of Yersinia pestis http://www.cdc.gov/ncidod/dvbid/plague/wayson.htm The Laboratory Manual of Plague Diagnostic Tests, 2000 Edition, published by the CDC and WHO, also has descriptions of several staining methods for Yersinia pestis. This book is probably restricted in distribution like the LRN Reference Procedures and is currently out of print. A quick search of the Internet using Google and A9 might give some interesting results. If I have time this week, I will perform a search and get back to you with what I find. Michael P. Owen, Regulatory Microbiologist FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov From David.Muskett <@t> RLC.NHS.UK Mon Sep 20 10:03:55 2004 From: David.Muskett <@t> RLC.NHS.UK (Muskett David) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Ehrlich's Haematoxylin[Scanned] Message-ID: <6AB79F0BA7C4914BA8DD8F5E0C21B97513BEEA@AHEXMAIL01.xalderhey.com> Dear All I am currently reviewing our Haematoxylin and Eosin staining which is not as strong as I would like. We are currently using Ehrlich's haematoxylin and would like to continue doing so. Does anyone have a recipe for Ehrlich's Haematoxylin that they can share with me. Thanks in advance David David Muskett Chief Biomedical Scientist, Histology Royal Liverpool Children's NHS Trust Eaton Road Liverpool L12 2AP Telephone (0151) 293 3656 Fax (0151) 293 3617 From Jenny-Oblander <@t> omrf.ouhsc.edu Mon Sep 20 10:09:59 2004 From: Jenny-Oblander <@t> omrf.ouhsc.edu (Jenny Oblander) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] yersinia pestis Message-ID: http://www.bt.cdc.gov/Agent/Plague/ype_la_cp_121301.pdf -----Original Message----- From: Jill Songer [mailto:jtsonger@vt.edu] Sent: Monday, September 20, 2004 9:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] yersinia pestis Does anyone out there in cyber land know the best stain for this bacteria? Thanks in advance. 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From j_gorenstein <@t> yahoo.com Mon Sep 20 10:51:00 2004 From: j_gorenstein <@t> yahoo.com (Juile Gorenstein) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] looking for Histologist (Novartis, Cambridge, MA) Message-ID: <20040920155100.85765.qmail@web42006.mail.yahoo.com> Hello everyone, If this looks interesting, please forward your resume to me and I will pass it along to my boss. Julie Description / Key Responsibilities: We are looking for a highly motivated histologist to work with pathologists in the Models of Disease Center in the Novartis Institute for Biomedical Research Inc. based in Cambridge, Massachusetts. The candidate is responsible for collecting and processing tissues from transgenic and knockout mice, histological and microscopic analyses, and data collection and documentation. The position offers an ideal opportunity to work with many talented scientists in a highly active and innovative research environment. Minimum Requirements: B.S. or M.S. in biology with minimum of three years of research experience in histology and microscopy. Experience with transgenic and knockout mouse models is desirable. Candidates should have experience in computer applications such as Microsoft Word, Excel, Powerpoint etc. and good communication skills. --------------------------------- Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. From histophilhuff <@t> yahoo.com Mon Sep 20 11:04:50 2004 From: histophilhuff <@t> yahoo.com (Phillip Huff) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Digital camera setup for histology teaching - Mac or PC? In-Reply-To: <000a01c49ed9$59a4c940$8c0fd680@ekk1116> Message-ID: <20040920160450.28341.qmail@web50302.mail.yahoo.com> Are you using a Mac or PC as your computer system? Phil Mikael Niku wrote: Dear Histonetters, we are planning to replace our old histology teaching equipment (an analog video camera attached to the teacher's microscope + monitors for the students) with new digital stuff (digital video camera + data projector). Please recommend a setup. I have the following requirements in my mind: - The idea is to display the view from the teacher's microscope on the screen. - We need LIVE image of course. - A computer software supporting the teaching would be nice (for example, allowing the teacher to draw on the live image, and perhaps showing several simultaneous images from the memory on the screen). - Max resolution of the projector is 1280x1024, so the camera doesn't need a superb resolution. Good color reproduction is necessary, though, as well as good automatic brightness adjustment. - We need to fit the camera on the old Olympus BH2. Thanks in advance. //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mitäkö mieltä olen länsimaisesta sivistyksestä? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. From JQB7 <@t> CDC.GOV Mon Sep 20 11:08:36 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] yersinia pestis Message-ID: Jill: Are you dealing with smears or tissue? If tissue, is it fresh or fixed? Jeanine Bartlett CDC -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Jill Songer Sent: Mon 9/20/2004 10:34 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] yersinia pestis Does anyone out there in cyber land know the best stain for this bacteria? Thanks in advance. 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Mon Sep 20 12:26:19 2004 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] dissecting pins Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5C071@EXCHANGE1.huntingtonhospital.com> Can anyone tell me where I can purchase T-shaped pins for pinning out specimens? Laurie Colbert Huntington Memorial Hospital Pasadena, CA From celebrej <@t> HHSC.CA Mon Sep 20 12:42:28 2004 From: celebrej <@t> HHSC.CA (Celebre Julia) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] dissecting pins Message-ID: <3AADFB88753AD31189C100902786B91C0E41CF61@hch_nt_exchange.hhsc.ca> We purchased ours in the sewing section of Wallmart... Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext46145 email: celebrej@hhsc.ca -----Original Message----- From: Laurie Colbert [mailto:laurie.colbert@huntingtonhospital.com] Sent: Monday, September 20, 2004 1:26 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] dissecting pins Can anyone tell me where I can purchase T-shaped pins for pinning out specimens? Laurie Colbert Huntington Memorial Hospital Pasadena, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 jluis.palazon <@t> icman.csic.es Mon Sep 20 13:25:28 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] hematoxylin Message-ID: <20040920182528.DD85B2D7472@perceval.uca.es> Dear List-members greetings. I would like to know if there is a substitute for the "celestine blue/hematoxylin" staining of nucleus that can resist a posterior acidic counterstain. I need to apply a protocol that uses this nuclear staining method but I dont have celestine blue and the company that suplies it says that it would last 2 months to supply the stain Thanks in advance Jos? Luis From kabrady <@t> genomichealth.com Mon Sep 20 13:52:05 2004 From: kabrady <@t> genomichealth.com (Kimberly Brady) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Gill Hematoxylin H&E Message-ID: <5C9B539E7DD26C4AA41D4E27ACECBB910197BE70@jaguar.genomichealth.com> Does anybody have an H&E protocol that calls for using Gill's Hematoxylin? Any help would be appreciated. Thanks, Kim The contents of this electronic message, including any attachments, are intended only for the use of the individual or entity to which they are addressed and may contain confidential information. If you are not the intended recipient, you are hereby notified that any use, dissemination, distribution, or copying of this message or any attachment is strictly prohibited. If you have received this transmission in error, please send an e-mail to postmaster@genomichealth.com and delete this message, along with any attachments, from your computer. From galinadeyneko <@t> yahoo.com Mon Sep 20 15:07:01 2004 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] cell culture Message-ID: <20040920200701.52880.qmail@web14525.mail.yahoo.com> Steve, web-site www.corning.com/lifesciences and find Preparation of Costar Transwell Inserts for Histology. Follow this protocol . I did it and received good results. call me if you have questions. Galina Deyneko Novartis 617-871-7613 --------------------------------- Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. From clarke.ian <@t> virgin.net Sat Sep 18 16:14:12 2004 From: clarke.ian <@t> virgin.net (ian clarke) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Re Uni/bi-directional communication with slide/cassette writers and LMS Message-ID: Hi all, I am looking for information regarding the new slide and cassette writers.I am very interested to see that the slide/cassette writers from Sakura and Leica can produce 2D bar codes on cassettes.I would like to know if the information can be communicated from the LMS to the Writers and from the writers to the LMS.For instance if a block is eventually levelled can the LMS be updated from the extra slides produced from the slide writer or do you have to imput the information into the LMS which then produces the extra slide labels?It would be of great adavntage if the information flow was bi-directional from the LMS and the Slide/cassette writers. Thanks Ian From VIKOHUT <@t> aol.com Mon Sep 20 16:46:42 2004 From: VIKOHUT <@t> aol.com (VIKOHUT@aol.com) Date: Fri Sep 16 15:24:03 2005 Subject: [Histonet] Clinical ladder in Histology Message-ID: <140.33569eae.2e80a9c2@aol.com> Dear Histonetters: In our laboratory the Med Techs have made a Clinical Ladder to provide individuals venues to increase their status (this includes monetary adjustment). We are now trying to implement this for the Histology Dept.. I was wondering if any other institution has this kind of Clinical Ladder for Histology or anything similar. All comments are appreciated. Thank you in advance Iryna Kohut HT ASCP Jersey Shore University Med Center Neptune NJ From katri <@t> cogeco.ca Mon Sep 20 20:40:13 2004 From: katri <@t> cogeco.ca (katri@cogeco.ca) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] hematoxylin Message-ID: <414f867d.104.74ae.2494@cogeco.ca> > Hi Jose, Try Weigert's iron hematoxylin. Katri Katri Tuomala St.Joseph's Health Care Hamilton, Ontario, Canada > Dear List-members > > greetings. I would like to know if there is a substitute for the "celestine blue/hematoxylin" staining of nucleus that > can resist a posterior acidic counterstain. I need to apply a protocol that uses this nuclear staining method but I > dont have celestine blue and the company that suplies it says that it would last 2 months to supply the stain > > Thanks in advance > > Jos? Luis > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bill501 <@t> mindspring.com Mon Sep 20 21:20:30 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Yersinia pestis Staining In-Reply-To: References: Message-ID: See also "Immunohistochemical Detection of Yersinia pestis in Formalin-Fixed, Paraffin-Embedded Tissue" Am J Clin Pathol 2002;117:205-209 I have a PDF of this if you are interested. Bill From meyenhmf <@t> umdnj.edu Mon Sep 20 18:51:12 2004 From: meyenhmf <@t> umdnj.edu (meyenhmf@umdnj.edu) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Glycerin-Gelatine bubble-phenomen In-Reply-To: <01cd01c49eee$91bc2c50$1401a8c0@GERICHTS9XOZZ8> References: <01cd01c49eee$91bc2c50$1401a8c0@GERICHTS9XOZZ8> Message-ID: <1095742272.414fb34026161@webmail.umdnj.edu> Bubbles are caused by desintigration of uncouppled, tissue bound dyes, a phenomenon described in the 60ties occuring with some enzyme histochemistry reactions. A treatment with acidic formalin and sulfanilic acid after the reaction would reduce/eliminate the bubbling effect. Regards, Markus F. Meyenhofer Quoting Barbara Bublava : > Hi Everybody! > > I have an interesting phenomen mounting slides with Glyceringelatine > > When I mount my slides they do not have bubbles. Oil Red O stains do not > develop bubbles even after some time but when I mount CAE - Chloracetate > Esterase (I mount in the same way I do the ORO) the slides develop > bubbles after some hours and after some days the whole section is covered > with bubbles, but there are no bubbles around the section! > > Why does this happen and how can I prevent this??? > > I use Glyceringelatine at 60?C, slides out of dest. water, excess water > shaked of. I did also try to dry the slides bevore mounting but there > happend to be more bubbles from the beginning and it did not change this > strange effect. > Sometimes, when I am in a hurry I further hardening of the > Glyceringelatine at fridge (4?C) but I can not see any positive or > negative effect on bubbles doing that. > > Thanks to everyone > > Barbara, Vienna > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- From Chewy71874 <@t> aol.com Tue Sep 21 01:58:46 2004 From: Chewy71874 <@t> aol.com (Chewy71874@aol.com) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Looking for histotech Message-ID: <128.4ba66dca.2e812b26@aol.com> Looking for an opportunity? Diagnostic Pathology Medical Group is looking for a trained histotechnician. ASCP licensure is preferred, but we will consider you if you are fresh out of training and ready to meet a challenge. Human and animal tissues. Competitive salary & excellent benefits. Fax resume to (916)447-0620 or call (916)447-2718. Ellen Yee Diagnostic Pathology Medical Group 2420 J Street Sacramento, CA 95816 Also, does anyone have the e-mail address of person(s) who list histotech jobs? Ellen From rentonlf <@t> bru.wits.ac.za Tue Sep 21 02:12:35 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] hematoxylin Message-ID: <1095750755.97eb86e0rentonlf@bru.wits.ac.za> I am not sure whose Haematoxylin this is, but we have used it extensively for many years for all trichrome stains. The recipe is is Bancroft & Stevens, should you need the ref. Stable Iron Haematoxylin: Soln A: 1g haem powder 100ml 95% alcohol. Stir until powder is dissolved Soln B: 10g Aluminium Chloride(hydrated)[AlCl3.6h2O) 10g Ferrous Sulphate(hydrated) 100ml Dist Water Stir until dissolved and add Soln A & B together. Then add: 2ml conc HCl 2ml 9% Na Iodate Mix & allow to stand at RT 48hrs before use. Soln remains stable at 4deg C for 2 months. Staining time 10 mins, rinse in water and differentiate in 1min in acid alcohol if required. regards -----Original Message----- From: Jose Luis Palazon Fernandez To: histonet@lists.utsouthwestern.edu Date: Mon, 20 Sep 2004 20:25:28 +0200 (CEST) Subject: [Histonet] hematoxylin Dear List-members greetings. I would like to know if there is a substitute for the "celestine blue/hematoxylin" staining of nucleus that can resist a posterior acidic counterstain. I need to apply a protocol that uses this nuclear staining method but I dont have celestine blue and the company that suplies it says that it would last 2 months to supply the stain Thanks in advance Jos? Luis _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From david.kinsley <@t> spcorp.com Tue Sep 21 09:23:40 2004 From: david.kinsley <@t> spcorp.com (Kinsley, David) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] solochrome cyanine-iron alum Message-ID: Does anyone out there have a procedure for, or have ever used this staining method? Any information is greatly appreciated. David Kinsley Schering-Plough Research Institute Kenilworth, NJ 07033 908-740-4669 ********************************************************************* 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 david.kinsley <@t> spcorp.com Tue Sep 21 09:21:57 2004 From: david.kinsley <@t> spcorp.com (Kinsley, David) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Calcein fluorescent staining of bone Message-ID: I have been asked to monitor bone growth in rats by administering Calcein by subcu. injection and would like to know if anyone has experience with this procedure, and if anyone could recommend the correct Calcein to purchase? Any assistance is appreciated. thanks David Kinsley Schering-Plough Research Institute Kenilworth, NJ 07033 908-740-4669 ********************************************************************* 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 STEGTM <@t> samcstl.org Tue Sep 21 12:13:14 2004 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:24:04 2005 Subject: [BULK] - Re: Fwd: Re: [Histonet] offtopic: histologist on "theBenefactor" Message-ID: Store it in a cool, dry place. I believe you mean the Wilburys. Terre >>> "Fred Underwood" 9/15/2004 2:31:23 PM >>> I have that CD. It has Tom Petty, Jeff Lynne, George Harrison, Bob Dylan and Roy Orbison. >>> Ford Royer 09/15/04 03:27PM >>> Kim Merriam wrote: >link to bio on "the Benefactor" web site. Not sure what a traveling histologist does. > > ....they gather no moss? maybe? ~ Ford Ford M. Royer M.S.B. Minneapolis, MN _______________________________________________ Histonet mailing list Histonet@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 psanquin <@t> lugo.usc.es Tue Sep 21 13:20:20 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Paraffin embedding Message-ID: <3.0.6.32.20040921202020.0080b320@pop.lugo.usc.es> Dear listers, After the summer break I am back to the list with my first lab headache of the season. The first thing I did was replacing all the liquids in the paraffin embedder, Hopefully the first embedding (pieces of mouse brain fixed in Paraformaldehyde 4%) should be nice, but when cutting the blocks in the microtome I have realized that they are dreadfully ruined. I think that the fixation is correct as I have processed pieces from the same animal with the vibratome and they seem fine. So probably something is wrong with the embedding. Perhaps with the new liquids should I decrease the time of the embedding specially the final paraffin steps. Have you got some hint? How long do you suggest? They are very small brain pieces from postanatal mice. Thre paraffin I use is Tissue-Tek III 4502 from SAKURA Have you got some input about it? Thanks in advance Pablo From contact <@t> excaliburpathology.com Tue Sep 21 12:45:43 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Gill Hematoxylin Message-ID: <20040921174543.13721.qmail@web50308.mail.yahoo.com> I use Gill III Hematoxylin on my eye sections. Stain 10 minutes in Gill III Rinse in tap water Differintiate in acid alcohol 3 dips Rinse in tap water Blue in dilute ammonia water for 1 minute Rinse in tap water for 1 min Proceed with counterstain From MinHan.Tan <@t> vai.org Tue Sep 21 14:15:55 2004 From: MinHan.Tan <@t> vai.org (Tan, MinHan) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Peptide preparation for antibody validation Message-ID: Good afternoon, fellow Histonetizens: - I would like to enquire as to the preparation of peptide for the purposes of antibody validation. - We have a rabbit polyclonal antibody against a novel peptide target made by Invitrogen that has cytoplasmic staining on IHC in formalin fixed tissue. - We do not have any more of that peptide, and we would like to make some to perform some competitive IHC (pre-incubate peptide + antibody) - I would like to enquire: what kind of purity of peptide do we need (70%, 90%, 95%?) and does it need to be amidated? - If there is anything else I'd need to watch out for, would appreciate any pointers! :) Thank you! Min-Han Tan Van Andel Institute 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 psanquin <@t> lugo.usc.es Tue Sep 21 14:37:41 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Paraffin embedding In-Reply-To: <000001c4a00a$ddfdc750$3601a8c0@brownpathology.net> References: <3.0.6.32.20040921202020.0080b320@pop.lugo.usc.es> Message-ID: <3.0.6.32.20040921213741.007da7b0@pop.lugo.usc.es> Dear Bonnie, The tissue is fractured showing numerous cracks or fissures always filled with paraffin. They are more conspicuous in the periphery but the whole section is useless. Thanks for your help. Pablo At 01:43 p.m. 21/09/04 -0500, you wrote: >Please give us details of exactly what is wrong. >Thanks, > >Bonnie Whitaker >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pablo >S?nchez Quinteiro >Sent: Tuesday, September 21, 2004 1:20 PM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Paraffin embedding > >Dear listers, > >After the summer break I am back to the list with my first lab headache of >the season. > >The first thing I did was replacing all the liquids in the paraffin >embedder, Hopefully the first embedding (pieces of mouse brain fixed in >Paraformaldehyde 4%) should be nice, but when cutting the blocks in the >microtome I have realized that they are dreadfully ruined. I think that the >fixation is correct as I have processed pieces from the same animal with >the vibratome and they seem fine. > >So probably something is wrong with the embedding. Perhaps with the new >liquids should I decrease the time of the embedding specially the final >paraffin steps. Have you got some hint? How long do you suggest? They are >very small brain pieces from postanatal mice. > >Thre paraffin I use is Tissue-Tek III 4502 from SAKURA Have you got some >input about it? > >Thanks in advance > >Pablo > > > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From tissuearray <@t> hotmail.com Tue Sep 21 14:54:52 2004 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Immuno tissues falling off... Message-ID: I was wondering what methods immuno folks are using to keep bone marrow sections on the slide? Glues or what? Our main problem is during pretreatment, the samples just fall off. I have tried to use 10% elmers glue on the slides. It helps for some tissues, bone marrow biopsys are not staying on at all. Anyone have any suggistions? Thanks, Thom _________________________________________________________________ [1]On the road to retirement? Check out MSN Life Events for advice on how to get there! References 1. http://g.msn.com/8HMAENUS/2749??PS=47575 From psanquin <@t> lugo.usc.es Tue Sep 21 14:37:41 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Paraffin embedding In-Reply-To: <000001c4a00a$ddfdc750$3601a8c0@brownpathology.net> References: <3.0.6.32.20040921202020.0080b320@pop.lugo.usc.es> Message-ID: <3.0.6.32.20040921213741.007da7b0@pop.lugo.usc.es> Dear Bonnie, The tissue is fractured showing numerous cracks or fissures always filled with paraffin. They are more conspicuous in the periphery but the whole section is useless. Thanks for your help. Pablo At 01:43 p.m. 21/09/04 -0500, you wrote: >Please give us details of exactly what is wrong. >Thanks, > >Bonnie Whitaker >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pablo >S?nchez Quinteiro >Sent: Tuesday, September 21, 2004 1:20 PM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Paraffin embedding > >Dear listers, > >After the summer break I am back to the list with my first lab headache of >the season. > >The first thing I did was replacing all the liquids in the paraffin >embedder, Hopefully the first embedding (pieces of mouse brain fixed in >Paraformaldehyde 4%) should be nice, but when cutting the blocks in the >microtome I have realized that they are dreadfully ruined. I think that the >fixation is correct as I have processed pieces from the same animal with >the vibratome and they seem fine. > >So probably something is wrong with the embedding. Perhaps with the new >liquids should I decrease the time of the embedding specially the final >paraffin steps. Have you got some hint? How long do you suggest? They are >very small brain pieces from postanatal mice. > >Thre paraffin I use is Tissue-Tek III 4502 from SAKURA Have you got some >input about it? > >Thanks in advance > >Pablo > > > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From stephen.asquith <@t> petermac.org Wed Sep 22 01:13:23 2004 From: stephen.asquith <@t> petermac.org (Asquith Stephen) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Making Ralph Knives Message-ID: Hi Histonetters, Question: Has anyone modified their LKB 7800 EM Knife maker to make Ralph knives? (I know that LKB actually made a dedicated Ralph maker however we don't really want to buy one) It appears to me that if you were to exchange the two contact points under the glass to a single strip contact i.e.. the crack would propagate from the score line on the top surface of the glass to an edge rather than two point contacts on the bottom edge of the glass. I have found that that without any modification I can make a reasonable knife, the with knife profile is OK but the cutting edge has two high points that correspond to the contacts. Any feed back would be much appreciated. Steve From wrhewett <@t> austarnet.com.au Wed Sep 22 04:02:44 2004 From: wrhewett <@t> austarnet.com.au (william hewett) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] tissues falling off Message-ID: <007601c4a082$edd10e30$70cadccb@hewett> Hi, Thom We find silanized slides from Dakocytomation will fix trephine marrow sections for immunostaining.Sections are placed in a sixty degree oven for 45 min. I have done microwave pressure cooker retreival on these sections with no loss. Bill QHPS Lab. Townsville.Aust. From Inga.Hansson <@t> neuro.uu.se Wed Sep 22 06:05:54 2004 From: Inga.Hansson <@t> neuro.uu.se (Inga Hansson) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] double stainings Message-ID: Hi everyone! If I want to do a double staining using one antibody that requires HIER and another that will stain everything if heated........what do I do? Can anyone help me? Thanks ! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 From tflore <@t> lsuhsc.edu Wed Sep 22 06:29:40 2004 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Making Ralph Knives Message-ID: Steve, our EM lab has three knife makers, LKB's and another and we even have a Ralph knife maker but with the workload and the time consuming preparing of glass knives we have gone to sectioning our "thick" sections (we use spurr epoxy) with the Diatome Histoknife, which have a larger cutting surface, can be reused as long as you do not have a knick, and even then they are guaranteed so many resharpenings by EMS. OK, enough of why we switched, good luck. Teresa Flores LSUHSC New Orleans, LA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Asquith Stephen Sent: Wednesday, September 22, 2004 1:13 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Making Ralph Knives Hi Histonetters, Question: Has anyone modified their LKB 7800 EM Knife maker to make Ralph knives? (I know that LKB actually made a dedicated Ralph maker however we don't really want to buy one) It appears to me that if you were to exchange the two contact points under the glass to a single strip contact i.e.. the crack would propagate from the score line on the top surface of the glass to an edge rather than two point contacts on the bottom edge of the glass. I have found that that without any modification I can make a reasonable knife, the with knife profile is OK but the cutting edge has two high points that correspond to the contacts. Any feed back would be much appreciated. Steve _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MAUGER <@t> email.chop.edu Wed Sep 22 07:02:47 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] double stainings Message-ID: Dear Inga, For double staining such markers, you can first stain the antibody that does not require heat. After the chromogen step, do your heat retrieval, and then stain with the next primary Ab. Jo >>> "Inga Hansson" 09/22/04 07:05AM >>> Hi everyone! If I want to do a double staining using one antibody that requires HIER and another that will stain everything if heated........what do I do? Can anyone help me? Thanks ! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Luis.Chiriboga <@t> med.nyu.edu Wed Sep 22 07:41:04 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] double stainings In-Reply-To: Message-ID: I have used Joanne's idea before, and it has worked quite well. the only problem I have observed is that the first chromogen sometimes looses it's "crispiness". So I would suggest that you be careful which enzyme/chromogen combination you use first. In my experience I have found that peroxidase/DAB is the least affected while alk-phosp/nbt-bcip is the worse. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne Mauger Sent: Wednesday, September 22, 2004 8:03 AM To: histonet@lists.utsouthwestern.edu; Inga.Hansson@neuro.uu.se Subject: Re: [Histonet] double stainings Dear Inga, For double staining such markers, you can first stain the antibody that does not require heat. After the chromogen step, do your heat retrieval, and then stain with the next primary Ab. Jo >>> "Inga Hansson" 09/22/04 07:05AM >>> Hi everyone! If I want to do a double staining using one antibody that requires HIER and another that will stain everything if heated........what do I do? Can anyone help me? Thanks ! Inga -- Inga Hansson dept. neuroscience, div. neurobiology PO Box 587 Biomedical Centre Husargatan 3 S-751 23 Uppsala SWEDEN phone:+46-18-4714384 fax: +46-18-559017 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Pathologyarts <@t> aol.com Wed Sep 22 08:45:18 2004 From: Pathologyarts <@t> aol.com (Pathologyarts@aol.com) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] who had the extra H. Pylori tissue Message-ID: I lost the email from the Physician who had some extra tissue, any help would be appreciated. Curt From RCazares <@t> schosp.org Wed Sep 22 09:14:56 2004 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Figuring cost per H&E slide Message-ID: <229A3566B9F0D311826E00D0B7441D79085C2E77@swedish_nt1.schosp.org> Hello Histonetters, How can I figure out cost per H&E slide including labor? Is there a formula that is used? I was just going to add up all the reagent costs for the last 2 months and divide it by the number of slides put out for those two months, but how do I factor in the tech time? I'd appreciate any help I can get. Thanks for any information, Ruth Cazares Swedish Covenant Hospital Chicago, IL *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. ************************* From DDittus787 <@t> aol.com Wed Sep 22 09:30:03 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Figuring cost per H&E slide Message-ID: <3E202735.4F042CB2.0A1F969F@aol.com> RUTH: You may want to look at how many slides are cut in say 4 hours and divide by your techs salary per hour. I do it on 15 min increments and total slides . for example if a tech gets 20.00/hr then 15 min is 5.00 and if she cuts 15 slides in that 15 min labor is 1.00/slide. a little rudimentary but effective. reagen costs should be based on total slides, before reagents changed. Do you change any reagents during daily runs? do you use hematoxylin for more that a day? if you use for 5 days you need to calculate that cost on 5 days worth of slides and actually makes your costs lower. if you need more help feel free to call me at work dana 215-481-2609 From STapper <@t> slhduluth.com Wed Sep 22 10:36:24 2004 From: STapper <@t> slhduluth.com (Tapper, Sheila) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Waterbath antigen retrieval Message-ID: <3BFBBD68413CB443A7125A63EC0ACD1D6716FD@slhw2smail01.slhdomain.com> For those of you that use the waterbath method for antigen retrieval, what waterbath do you use? I would appreciate manufacturer information. We have burned through 2 in the last 6 months - obviously we need to find something different. Thanks for the information! Sheila Tapper HT(ASCP) St. Luke's Hospital Duluth, MN 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 info <@t> instrumedics.com Wed Sep 22 12:19:43 2004 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Re:tissue falling off the slide References: Message-ID: <012501c4a0c8$5a2787e0$6401a8c0@INSTRUMEDICS22> Thom, Many people find the best way to secure a bone section to the slide is using tape-transfer system where the section is adhered to an adhesive-coated slide. The adhesive is converted to a plastic and the section is anchored to it and will stay on the slide even in very rigorous protocols Bernice ----- Original Message ----- From: To: Sent: Wednesday, September 22, 2004 1:02 PM Subject: Histonet Digest, Vol 10, Issue 29 > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > ---------------------------------------------------------------------------- ---- > Today's Topics: > > 1. [BULK] - Re: Fwd: Re: [Histonet] offtopic: histologist on > "theBenefactor" (Therersa Stegall) > 2. Paraffin embedding (Pablo S?nchez Quinteiro) > 3. Gill Hematoxylin (Paula Pierce) > 4. Peptide preparation for antibody validation (Tan, MinHan) > 5. RE: Paraffin embedding (Pablo S?nchez Quinteiro) > 6. Immuno tissues falling off... (Thom Jensen) > 7. RE: Paraffin embedding (Pablo S?nchez Quinteiro) > 8. Making Ralph Knives (Asquith Stephen) > 9. tissues falling off (william hewett) > 10. double stainings (Inga Hansson) > 11. RE: Making Ralph Knives (Flores, Teresa) > 12. Re: double stainings (Joanne Mauger) > 13. RE: double stainings (Luis Chiriboga) > 14. who had the extra H. Pylori tissue (Pathologyarts@aol.com) > 15. Figuring cost per H&E slide (Cazares, Ruth) > 16. Re: Figuring cost per H&E slide (DDittus787@aol.com) > 17. Waterbath antigen retrieval (Tapper, Sheila) > ---------------------------------------------------------------------------- ---- > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Wed Sep 22 13:31:38 2004 From: carl.hobbs <@t> kcl.ac.uk (carl hobbs) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] double stainings Message-ID: I agree with previous two posts. DAB for chromogen of non-HIER, then whatever you want to contrast for second Ag. Technically one should use a different enzyme system but, if the Ags are not co-localised, I might do HRP/DAB/Cobalt/Nickel enhanced, giving brown and blue/black. The first reporter molecule is sufficiently masked for it not to interfere. Obviously do parallel sections with individual detection systems just for your/their peace of mind. Only need to do that once . --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.762 / Virus Database: 510 - Release Date: 13/09/2004 From pzeitlow <@t> bbpllab.com Wed Sep 22 13:43:36 2004 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] rcc Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A0BEA85@bbplsrv1.bbpl> I am looking to evaluate RCC (renal cell carcinoma) antibodies. Is anyone willing to share the clone, vendor and experiences for this antibody. I will be using it on paraffin embedded tissues. TIA. Pat From rkrug <@t> sial.com Wed Sep 22 13:57:28 2004 From: rkrug <@t> sial.com (Robert Krug) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Long term storage of tissue Message-ID: I have a question concerning the long term storage of tissue which is not addressed in any of the histology textbooks I have on hand. After many hours of searching, I was unable to find an answer through a websearch. Can tissue be preserved in 10% neutral buffered formalin for decades without changing the 10% NBF (provided the container is sealed and does not leak)? A former coworker of mine once worked at the CDC and she stated the CDC used 70% alcohol for the long term storage of tissue. A review of the Internet shows that the Australian Museum Fish site states they have fixed in "formaldehyde and then stored in alcohol" for the last 80 years. This site claims to have thousands of specimens on hand. Another site I found recommended storage in formalin as a way to keep the original color of the tissue and recommended against long term storage of tissue in alcohol for various reasons. The unused 10% neutral buffered formalin solution does decompose over time and is typically assigned an expiration period. Once the tissue has been fixed in 10% neutral buffered formalin, is the reaction stable, meaning the bonds are stable and no decomposition will occur? I was always told the formalin solution degrades with time and may form formic acid and other by products. Given enough time the remaining formalin in solution might form formic acid, drop out of solution as paraformaldehyde or escape as formaldehyde gas. Another coworker feels that once fixation has occurred, the fixation is permanent and there is no need to change the solution. I realize the fixation bonds can be broken through washing in running water and with select chemicals. But given a sealed container, will the bonds remain permanent and the tissue free of decomposition or fungal growth? Any jounal articles or textbook references you have to support your view point would be greatly appreciated. Many thanks Bob Krug St John, Missouri From sharon.osborn <@t> dnax.org Wed Sep 22 14:12:33 2004 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Salmonella staining Message-ID: <29B25753F6B1D51196110002A589D44401A17813@PALMSG30.us.schp.com> Does anyone know or use other technics (in addition to Gram stain) for Salmonella bacterium in paraffin sections? I know the Gram stain is the standard staining technic used. Are there other histochemical technics that will also work? What about IHC? I appreciate your input. Sharon Osborn DNAX Palo Alto, CA 650.496.6539 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From TJasper <@t> smdc.org Wed Sep 22 14:26:38 2004 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Long term storage of tissue Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E44FBA@harrier> Robert, There may be some folks on this list from Mayo that can help you out. I know they have tissue stored there that is from the 19th Century. Also, the Russians have the body of Lenin which has been preserved for decades (under orders of the old Soviet government). A search on Google may help you out there. Good luck. Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org -----Original Message----- From: Robert Krug [mailto:rkrug@sial.com] Sent: Wednesday, September 22, 2004 1:57 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Long term storage of tissue I have a question concerning the long term storage of tissue which is not addressed in any of the histology textbooks I have on hand. After many hours of searching, I was unable to find an answer through a websearch. Can tissue be preserved in 10% neutral buffered formalin for decades without changing the 10% NBF (provided the container is sealed and does not leak)? A former coworker of mine once worked at the CDC and she stated the CDC used 70% alcohol for the long term storage of tissue. A review of the Internet shows that the Australian Museum Fish site states they have fixed in "formaldehyde and then stored in alcohol" for the last 80 years. This site claims to have thousands of specimens on hand. Another site I found recommended storage in formalin as a way to keep the original color of the tissue and recommended against long term storage of tissue in alcohol for various reasons. The unused 10% neutral buffered formalin solution does decompose over time and is typically assigned an expiration period. Once the tissue has been fixed in 10% neutral buffered formalin, is the reaction stable, meaning the bonds are stable and no decomposition will occur? I was always told the formalin solution degrades with time and may form formic acid and other by products. Given enough time the remaining formalin in solution might form formic acid, drop out of solution as paraformaldehyde or escape as formaldehyde gas. Another coworker feels that once fixation has occurred, the fixation is permanent and there is no need to change the solution. I realize the fixation bonds can be broken through washing in running water and with select chemicals. But given a sealed container, will the bonds remain permanent and the tissue free of decomposition or fungal growth? Any jounal articles or textbook references you have to support your view point would be greatly appreciated. Many thanks Bob Krug St John, Missouri _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Wed Sep 22 14:37:44 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] rcc Message-ID: I tried Novocastra's RCC-1 with no staining at all. I even used several different retrieval procedures. I heard that DAKO's is pretty good. Can you share your search results with me. Good luck. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 -----Original Message----- From: Pat Zeitlow [mailto:pzeitlow@bbpllab.com] Sent: Wednesday, September 22, 2004 1:44 PM To: histonet@pathology.swmed.edu Subject: [Histonet] rcc I am looking to evaluate RCC (renal cell carcinoma) antibodies. Is anyone willing to share the clone, vendor and experiences for this antibody. I will be using it on paraffin embedded tissues. TIA. Pat _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Wed Sep 22 14:41:47 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Salmonella staining Message-ID: I have a polyclonal antibody that I have used for the IHC detection of Salmonella. 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 >>> "Osborn, Sharon" 09/22/04 03:12PM >>> Does anyone know or use other technics (in addition to Gram stain) for Salmonella bacterium in paraffin sections? I know the Gram stain is the standard staining technic used. Are there other histochemical technics that will also work? What about IHC? I appreciate your input. Sharon Osborn DNAX Palo Alto, CA 650.496.6539 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Luis.Chiriboga <@t> med.nyu.edu Wed Sep 22 17:07:06 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] rcc In-Reply-To: Message-ID: RCC from novacastra is finicky. I went through the same process, trying several HIER methods as well as enzymes. Finally got it to work using a 1% trypsin for 30 minutes at 37C.....can send you pictures if interested (contact me off the list) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of GUTIERREZ, JUAN Sent: Wednesday, September 22, 2004 3:38 PM To: Pat Zeitlow; histonet@pathology.swmed.edu Subject: RE: [Histonet] rcc I tried Novocastra's RCC-1 with no staining at all. I even used several different retrieval procedures. I heard that DAKO's is pretty good. Can you share your search results with me. Good luck. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 -----Original Message----- From: Pat Zeitlow [mailto:pzeitlow@bbpllab.com] Sent: Wednesday, September 22, 2004 1:44 PM To: histonet@pathology.swmed.edu Subject: [Histonet] rcc I am looking to evaluate RCC (renal cell carcinoma) antibodies. Is anyone willing to share the clone, vendor and experiences for this antibody. I will be using it on paraffin embedded tissues. TIA. Pat _______________________________________________ Histonet mailing list Histonet@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 Wed Sep 22 17:55:19 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] rcc Message-ID: We are currently using clone "PN-15" from NeoMarkers/Lab Vision Corporation in Fremont, CA. I like it much better than other clones that we have tried. 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 >>> "Pat Zeitlow" 09/22/04 02:43PM >>> I am looking to evaluate RCC (renal cell carcinoma) antibodies. Is anyone willing to share the clone, vendor and experiences for this antibody. I will be using it on paraffin embedded tissues. TIA. Pat _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From antje.marcantonio <@t> pharma.novartis.com Thu Sep 23 02:01:14 2004 From: antje.marcantonio <@t> pharma.novartis.com (antje.marcantonio@pharma.novartis.com) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Waterbath antigen retrieval Message-ID: we use a waterbath from Fisher Scientific (Isotemp) and are happy with it BUT: it is only 2 years old right now and not in use every day advantage: light, therefore easy to change the water; quick heating and constant temperature Antje Marcantonio Novartis Pharma AG BU Transplantation Research Basel, Switzerland From Zorbutt <@t> aol.com Thu Sep 23 02:10:55 2004 From: Zorbutt <@t> aol.com (Zorbutt@aol.com) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] hsv-2 IHC antibody Message-ID: Dear all Has anyone ever used a herpes simplex virus-2 IHC antibody? If so do you know if the virus can be detected in its latent form? Thanx Zohra Butt From rentonlf <@t> bru.wits.ac.za Thu Sep 23 02:06:30 2004 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Long term storage of tissue Message-ID: <1095923190.8d5cbd80rentonlf@bru.wits.ac.za> Hmm, interesting question, this may not be an answer to your direct query, but from personal expereince I once tried to do PCR on FFPE blocks that were about 50 years old and found that the DNA was largely degraded. There are of course many explanations to this, one being that the formalin used way back then might not have been properly buffered (one favourite technique was to throw a handful of marble chips in the bottom ofthe 10% formalin container, so my assumption remains however that SOME degradation of SOME tissue factors may occur. -----Original Message----- From: Robert Krug To: histonet@lists.utsouthwestern.edu Date: Wed, 22 Sep 2004 13:57:28 -0500 Subject: [Histonet] Long term storage of tissue I have a question concerning the long term storage of tissue which is not addressed in any of the histology textbooks I have on hand. After many hours of searching, I was unable to find an answer through a websearch. Can tissue be preserved in 10% neutral buffered formalin for decades without changing the 10% NBF (provided the container is sealed and does not leak)? A former coworker of mine once worked at the CDC and she stated the CDC used 70% alcohol for the long term storage of tissue. A review of the Internet shows that the Australian Museum Fish site states they have fixed in "formaldehyde and then stored in alcohol" for the last 80 years. This site claims to have thousands of specimens on hand. Another site I found recommended storage in formalin as a way to keep the original color of the tissue and recommended against long term storage of tissue in alcohol for various reasons. The unused 10% neutral buffered formalin solution does decompose over time and is typically assigned an expiration period. Once the tissue has been fixed in 10% neutral buffered formalin, is the reaction stable, meaning the bonds are stable and no decomposition will occur? I was always told the formalin solution degrades with time and may form formic acid and other by products. Given enough time the remaining formalin in solution might form formic acid, drop out of solution as paraformaldehyde or escape as formaldehyde gas. Another coworker feels that once fixation has occurred, the fixation is permanent and there is no need to change the solution. I realize the fixation bonds can be broken through washing in running water and with select chemicals. But given a sealed container, will the bonds remain permanent and the tissue free of decomposition or fungal growth? Any jounal articles or textbook references you have to support your view point would be greatly appreciated. Many thanks Bob Krug St John, Missouri _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa .......so what IS the speed of dark? From kwuny <@t> email.cs.nsw.gov.au Thu Sep 23 02:55:39 2004 From: kwuny <@t> email.cs.nsw.gov.au (Young Kwun) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Collagen IV subtypes Message-ID: <200409231751338.SM05496@crgcsls814> Dear all, Does anyone know where I can get antibodies to subtypes of all collagen IV chains (alpha 1?alpha 6)? Are they only available for immunofluorescence detection? SantaCruz has some antibodies for collagen IV alpha 1, alpha 2 and alpha 3 isoforms. Thank you. Young Young Kwun 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 From M.J.Vonk <@t> lumc.nl Thu Sep 23 03:45:07 2004 From: M.J.Vonk <@t> lumc.nl (Vonk, M.J. (KNO)) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] re: cell culture Message-ID: Hello Galina and histonetters, I also tried to do histology on transwell inserts by Costar. I also followed the protocol but it did not work for me. The problem is that the paraffin does not stick to the membrane. What I got when I removed the membrane plug was that the membrane got separated from the paraffin. My cells are now partially on the membrane and partially in the paraffin plug. Has anyone got some ideas about how to embed such a transwell insert without destroying my cells? Marcel Vonk ******************************************* M.J.Vonk Leiden University Medical Center Dept of Otorhinolaryngology Room J2-77 Albinusdreef 2 PO-Box 9600 2300 RC Leiden The Netherlands tel: ..3171-5261179 M.J.Vonk@lumc.nl Message: 5 Date: Mon, 20 Sep 2004 13:07:01 -0700 (PDT) From: Galina Deyneko Subject: [Histonet] cell culture To: Stephen.Eyres@sanofi-synthelabo.com Cc: histonet@lists.utsouthwestern.edu Message-ID: <20040920200701.52880.qmail@web14525.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Steve, web-site www.corning.com/lifesciences and find Preparation of Costar Transwell Inserts for Histology. Follow this protocol . I did it and received good results. call me if you have questions. Galina Deyneko Novartis 617-871-7613 From c.m.vanderloos <@t> amc.uva.nl Thu Sep 23 05:45:49 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Re: double stainings Message-ID: <1412c0141d7a.141d7a1412c0@amc.uva.nl> Dear Inga and Luis, The approach of staining the 1st antigen including enzyme visualization, antigen retrieval and than the 2nd, would be also my advise in this case. However, when using DAB as chromogen for the first staining sequence please realize that this enzymatic product may shelter your 2nd antigen very effectively! This sheltering is not solved by the antigen retrieval step. Instead of DAB also Dako Permanent Red (Alk. phosp.) or X-gal (b-galactosidase) survives the antigen retrieval step quite well, whereas those chromogens doesn't have this sheltering characteristic. Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Luis Chiriboga Date Wed, 22 Sep 2004 08:41:04 -0400 To Joanne Mauger , histonet@lists.utsouthwestern.edu, Inga.Hansson@neuro.uu.se Subject RE: [Histonet] double stainings I have used Joanne's idea before, and it has worked quite well. the only problem I have observed is that the first chromogen sometimes looses it's "crispiness". So I would suggest that you be careful which enzyme/chromogen combination you use first. In my experience I have found that peroxidase/DAB is the least affected while alk-phosp/nbt-bcip is the worse. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne Mauger Sent: Wednesday, September 22, 2004 8:03 AM To: histonet@lists.utsouthwestern.edu; Inga.Hansson@neuro.uu.se Subject: Re: [Histonet] double stainings Dear Inga, For double staining such markers, you can first stain the antibody that does not require heat. After the chromogen step, do your heat retrieval, and then stain with the next primary Ab. Jo >>> "Inga Hansson" 09/22/04 07:05AM >>> Hi everyone! If I want to do a double staining using one antibody that requires HIER and another that will stain everything if heated........what do I do? Can anyone help me? Thanks ! Inga From Ianbernard <@t> netzero.com Thu Sep 23 06:36:00 2004 From: Ianbernard <@t> netzero.com (Ian) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Need Help with obtaining Staining Method- Demonstrates 5'-Nase enzyme activity and Alkaline Phosphatase Seining for Blood Capillaries Message-ID: I am looking for the Metal Salt method of Wachstein & Meisel modification by Heusermann. Also need the "Azo dye method of Burstone by Kato et al. I need steps and reagents and source of purchase. Please reply back to my both email addresses above. Thanks, SSgt Ian Bernard HT(ASCP) From Luis.Chiriboga <@t> med.nyu.edu Thu Sep 23 07:48:35 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] RE: double stainings In-Reply-To: <1412c0141d7a.141d7a1412c0@amc.uva.nl> Message-ID: I absolutely agree Chris, and I stopped using NBT-BCIP and have switched to Fast Red! I was wondering, do you know of any experimental rather than empirical evidence for the shielding effect? Empirically speaking, alk phosp may be more sensitive (I believe it has a lower detection limit than DAB.....based on western blotting?)than peroxidase/DAB, but the perox/DAB combo produces a superior signal in FF-PE. I have only limited experience with the x-gal detection systems. -----Original Message----- From: C.M. van der Loos [mailto:c.m.vanderloos@amc.uva.nl] Sent: Thursday, September 23, 2004 6:46 AM To: histonet@lists.utsouthwestern.edu Cc: Luis.Chiriboga@med.nyu.edu; Inga.Hansson@neuro.uu.se Subject: Re: double stainings Dear Inga and Luis, The approach of staining the 1st antigen including enzyme visualization, antigen retrieval and than the 2nd, would be also my advise in this case. However, when using DAB as chromogen for the first staining sequence please realize that this enzymatic product may shelter your 2nd antigen very effectively! This sheltering is not solved by the antigen retrieval step. Instead of DAB also Dako Permanent Red (Alk. phosp.) or X-gal (b-galactosidase) survives the antigen retrieval step quite well, whereas those chromogens doesn't have this sheltering characteristic. Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Luis Chiriboga Date Wed, 22 Sep 2004 08:41:04 -0400 To Joanne Mauger , histonet@lists.utsouthwestern.edu, Inga.Hansson@neuro.uu.se Subject RE: [Histonet] double stainings I have used Joanne's idea before, and it has worked quite well. the only problem I have observed is that the first chromogen sometimes looses it's "crispiness". So I would suggest that you be careful which enzyme/chromogen combination you use first. In my experience I have found that peroxidase/DAB is the least affected while alk-phosp/nbt-bcip is the worse. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne Mauger Sent: Wednesday, September 22, 2004 8:03 AM To: histonet@lists.utsouthwestern.edu; Inga.Hansson@neuro.uu.se Subject: Re: [Histonet] double stainings Dear Inga, For double staining such markers, you can first stain the antibody that does not require heat. After the chromogen step, do your heat retrieval, and then stain with the next primary Ab. Jo >>> "Inga Hansson" 09/22/04 07:05AM >>> Hi everyone! If I want to do a double staining using one antibody that requires HIER and another that will stain everything if heated........what do I do? Can anyone help me? Thanks ! Inga From jessica_butler <@t> oz.ped.emory.edu Thu Sep 23 08:36:29 2004 From: jessica_butler <@t> oz.ped.emory.edu (Jessica Butler) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Iron Stain Message-ID: Hi- I work at Emory University and we are currently investigating sickle cell disease. I need a protocol for staining mouse lung tissue for iron. The protocol and/or stain can be for paraffin fixed or frozen sections. Thanks- From portera203 <@t> yahoo.com Thu Sep 23 09:30:32 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] hsv-2 IHC antibody In-Reply-To: Message-ID: <20040923143032.53292.qmail@web40902.mail.yahoo.com> Zohra - I know that Nova Castra makes Herpes Simplex Virus I & II not sure about what form it detects though. They do have most of their specifications sheets on line that you can review. Zorbutt@aol.com wrote:Dear all Has anyone ever used a herpes simplex virus-2 IHC antibody? If so do you know if the virus can be detected in its latent form? Thanx Zohra Butt _______________________________________________ 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!? Yahoo! Mail - 50x more storage than other providers! From Cathy.Stevens <@t> HealthONEcares.com Thu Sep 23 10:15:34 2004 From: Cathy.Stevens <@t> HealthONEcares.com (Stevens Cathy) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Figuring cost per H&E slide Message-ID: That's how I have done it. -----Original Message----- From: Cazares, Ruth [mailto:RCazares@schosp.org] Sent: Wednesday, September 22, 2004 8:15 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Figuring cost per H&E slide Hello Histonetters, How can I figure out cost per H&E slide including labor? Is there a formula that is used? I was just going to add up all the reagent costs for the last 2 months and divide it by the number of slides put out for those two months, but how do I factor in the tech time? I'd appreciate any help I can get. Thanks for any information, Ruth Cazares Swedish Covenant Hospital Chicago, IL *** 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 Chewy71874 <@t> aol.com Thu Sep 23 10:47:51 2004 From: Chewy71874 <@t> aol.com (Chewy71874@aol.com) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Re: tissues falling off Message-ID: Thom, We use DAKOcytomation silanized slides. You also want to make sure when cutting the sections that you use DI or distilled water in the waterbath and no additives. Ellen Yee From jluis.palazon <@t> icman.csic.es Thu Sep 23 11:39:35 2004 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] question about inmuno Message-ID: <20040923163935.BDE432EB7DC@perceval.uca.es> Dear List-Members First I want to thank the people who kindly answered to my question about acid-resistant haematoxylin. I have a question about inmunohistochemistry. I have to make a inmuno to detect ATPase but then I would like to make another staining like PAS or H&E. Could these "counterstainins" affect the inmuno reaction or even cuould the inmuno response be lost or masked?. I want to distinguish to kinds of cells, both of then are positive to PAS but only one is positive to ATPase, I would like to show the two in the same picture. Any help or comentary would be appreciated thanks in advance Jos? Luis El dia 20/09/2004 21:13 usted envio el siguiente mensaje: >Date: 20 de Septiembre de 2004 21:13:32 >From: "Gudrun Lang" >Subject: Re: [Histonet] hematoxylin >To: jluis.palazon@icman.csic.es > > I think the usual substitute here is Weigert's Hematoxylin. It stains the > nuclei black. We use it at the beginning of Trichrom-stains. > staining: 5 min (or more), differentiation with 0,5% HCl-Alk. (obtional), > blueing 5 min tapwater. > > working solution: > Gieson A + Gieson B 1:1 (stable for one week, older solutions stain rather > brownish than black) > > Gieson A: > 10 g hematoxylin + 1000 ml 96% Alk. (allow to stand for one week) > > Gieson B: > 40 ml 29% Ferric-chlorid > 980 ml Aqua dest > 10 ml 25% HCl > > greetings > Gudrun > > > ----- Original Message ----- > From: "Jose Luis Palazon Fernandez" > To: > Sent: Monday, September 20, 2004 8:25 PM > Subject: [Histonet] hematoxylin > > > Dear List-members > > greetings. I would like to know if there is a substitute for the "celestine > blue/hematoxylin" staining of nucleus that can resist a posterior acidic > counterstain. I need to apply a protocol that uses this nuclear staining > method but I dont have celestine blue and the company that suplies it says > that it would last 2 months to supply the stain > > Thanks in advance > > Jos? Luis > > _______________________________________________ > 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 RCazares <@t> schosp.org Thu Sep 23 11:35:32 2004 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Thanks for info on H&E cost per slide Message-ID: <229A3566B9F0D311826E00D0B7441D79085C3010@swedish_nt1.schosp.org> Thank you all for your information. What was being asked of me was specifically was reagent cost and tech labor. This is what I did: I figured out how much alcohol, xylene, hematoxylin, eosin and clear-rite was used every week for H&E staining, translated that into dollars per week. Then I divided the total number of slides for the month by 4 weeks, (some weeks are slower than others, that's why I used the monthly total divided by 4 weeks). I then divided the reagent cost (weekly) by the number of slides (weekly), and this gave me a per slide total. To figure out the tech labor, I figured out how many actual hours are spent producing H&E slides per week and divided the weekly slide total by this. Then I took this total (slides per hour ) and then I divided the tech average salary per hour by the slides per hour and this gave me a per slide cost of tech labor. I'm sure there must be an easier way but this worked for me and I now have a nice chart in EXCEL showing all the numbers for future reference. If anyone sees a flaw in the way I did this please don't hesitate to let me know. Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. ************************* From histophilhuff <@t> yahoo.com Thu Sep 23 12:05:41 2004 From: histophilhuff <@t> yahoo.com (Phillip Huff) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Dissecting + Embedding Mouse Mammary Gland - Technique or Reference Message-ID: <20040923170541.56091.qmail@web50306.mail.yahoo.com> We will be beginning a new procedure of analyzing protein expression in mouse mammary gland, dissected immediately after the mother gives birth. We would like to know if anyone has an established protocol for dissecting the mammary glands from the mice and the subsequent procedure for embedding them for cryosectioning. I would also appreciate it if anyone has a recommended reference/journal article that outlines these procedures. Thanks for your knowledge, Phil __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From ploykasek <@t> phenopath.com Thu Sep 23 12:53:13 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Validation procedure In-Reply-To: Message-ID: While I do primarily IHC/ISH, I think the validation would be similar. I don't believe that a vendor can do your validation. The vendor can validate that their instrument is performing per it's specs, but that is NOT the same as validating the test. To validate the test you need to prove sensitivity & specificity with your patient specimens. This involves knowing what this test should & should not be positive on, having references to this pattern of staining, running a series of both positive & negative cases, writing this up explaining your results, etc... Then the test is validated for your facility with your patient population. Just my thoughts on the subject. Patti Loykasek PhenoPath Laboratories Seattle, WA > We recently went from manual special stains to automated special stains. We > correlated every stain with 3-5 old cases and one pathologist signed off on > these. Things have been so hectic these last several months that a > validation procedure has not been written down. I had read recently in CAP > that this needs written documentation, etc. Does anyone have a validation > procedure that they would be willing to share? I would be most grateful. > Thanks!!! > Paula Wilder > St. Joseph Medical Center > Towson, MD 21204 > > _________________________________________________________________ >> From ?will you?? to ?I do,? MSN Life Events is your resource for Getting > Married. http://lifeevents.msn.com/category.aspx?cid=married > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Thu Sep 23 12:57:58 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] RE: Long term storage of tissue Message-ID: <6229366a99.66a9962293@amc.uva.nl> Dear Robert, I don't think that there is an endpoint in NBF fixation. To my opinion the cross-linking will go on and on. We had a heart collection in NBF dating back to 1970. From time to time there was a IHC request on those old samples. Well, before the description of antigen retrieval (1992) it was fully impossible to stain anything except for some weak staining with anti-von Willebrand factor (after pepsin treatment of the sections). With antigen retrieval, it appeared that it was fully antibody dependent whether or not reliable staining was obtained. Also when there was a request of a series of specimens ranging from 1970 up to recent years, there was an increasing staining intensity of that antibody. Most successful was anti-alpha actin, clone 1A4 for staining smooth muscle cells. I think that one should be able to stain SMC's in mummy tissue! But detection of T-cells by CD3, CD8, etc. was totally impossible in those old specimens. However, the introduction of an ultra sensitive de tection system based on tyramide amplification definitely improved the situation. CD3, CD8 can be detected now by tyramide visualization in 10-20 year old specimens. I agree with you there is not much literature about this subject. Hope this opinion helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Robert Krug Date Wed, 22 Sep 2004 13:57:28 -0500 To histonet@lists.utsouthwestern.edu Subject [Histonet] Long term storage of tissue I have a question concerning the long term storage of tissue which is not addressed in any of the histology textbooks I have on hand. After many hours of searching, I was unable to find an answer through a websearch. Can tissue be preserved in 10% neutral buffered formalin for decades without changing the 10% NBF (provided the container is sealed and does not leak)? A former coworker of mine once worked at the CDC and she stated the CDC used 70% alcohol for the long term storage of tissue. A review of the Internet shows that the Australian Museum Fish site states they have fixed in "formaldehyde and then stored in alcohol" for the last 80 years. This site claims to have thousands of specimens on hand. Another site I found recommended storage in formalin as a way to keep the original color of the tissue and recommended against long term storage of tissue in alcohol for various reasons. The unused 10% neutral buffered formalin solution does decompose over time and is typically assigned an expiration period. Once the tissue has been fixed in 10% neutral buffered formalin, is the reaction stable, meaning the bonds are stable and no decomposition will occur? I was always told the formalin solution degrades with time and may form formic acid and other by products. Given enough time the remaining formalin in solution might form formic acid, drop out of solution as paraformaldehyde or escape as formaldehyde gas. Another coworker feels that once fixation has occurred, the fixation is permanent and there is no need to change the solution. I realize the fixation bonds can be broken through washing in running water and with select chemicals. But given a sealed container, will the bonds remain permanent and the tissue free of decomposition or fungal growth? Any jounal articles or textbook references you have to support your view point would be greatly appreciated. Many thanks Bob Krug St John, Missouri From contact <@t> excaliburpathology.com Thu Sep 23 12:57:53 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Long term storage of tissue Message-ID: <20040923175753.33476.qmail@web50309.mail.yahoo.com> Leaving tissues in 10% NBF will cause a black formalin pigment to precipitate in the tissues. Although the pigment can be removed, I store the wet tissue from the eyes I receive in 70% alcohol for years with no ill effect. From contact <@t> excaliburpathology.com Thu Sep 23 13:04:32 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Water bath antigen retrieval Message-ID: <20040923180432.36977.qmail@web50309.mail.yahoo.com> I use a Black and Decker rice steamer for antigen retrieval. About $30. Fill the bottom with distilled water. Place the plastic basket in the steamer. Place a container with your slides and citrate buffer in the basket so the lid will fit. Turn the steamer on for 1 hour, then let cool for 25 minutes. Rinse in distilled water slowly to finish cooling. Proceed with your IHC stain. From Cathy.Stevens <@t> HealthONEcares.com Thu Sep 23 13:30:44 2004 From: Cathy.Stevens <@t> HealthONEcares.com (Stevens Cathy) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Thanks for info on H&E cost per slide Message-ID: I even figure in the cost of mounting media, slides and coverslips -----Original Message----- From: Cazares, Ruth [mailto:RCazares@schosp.org] Sent: Thursday, September 23, 2004 10:36 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Thanks for info on H&E cost per slide Thank you all for your information. What was being asked of me was specifically was reagent cost and tech labor. This is what I did: I figured out how much alcohol, xylene, hematoxylin, eosin and clear-rite was used every week for H&E staining, translated that into dollars per week. Then I divided the total number of slides for the month by 4 weeks, (some weeks are slower than others, that's why I used the monthly total divided by 4 weeks). I then divided the reagent cost (weekly) by the number of slides (weekly), and this gave me a per slide total. To figure out the tech labor, I figured out how many actual hours are spent producing H&E slides per week and divided the weekly slide total by this. Then I took this total (slides per hour ) and then I divided the tech average salary per hour by the slides per hour and this gave me a per slide cost of tech labor. I'm sure there must be an easier way but this worked for me and I now have a nice chart in EXCEL showing all the numbers for future reference. If anyone sees a flaw in the way I did this please don't hesitate to let me know. Ruth Cazares *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. ************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TMcNemar <@t> lmhealth.org Fri Sep 24 06:35:49 2004 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Water bath antigen retrieval Message-ID: <90092A4ED388D7119575006008F7112049CBDE@NT_EXCHANGE> Wondering if this is your standard procedure for all antibodies? I only steam for 20 minutes with a 20 minute cool for all retrieval. Place them in buffer for 5 minutes then directly on the Autostainer. They work beautifully. Just curious, Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Paula Pierce [mailto:contact@excaliburpathology.com] Sent: Thursday, September 23, 2004 2:05 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Water bath antigen retrieval I use a Black and Decker rice steamer for antigen retrieval. About $30. Fill the bottom with distilled water. Place the plastic basket in the steamer. Place a container with your slides and citrate buffer in the basket so the lid will fit. Turn the steamer on for 1 hour, then let cool for 25 minutes. Rinse in distilled water slowly to finish cooling. Proceed with your IHC stain. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From japoteete <@t> saintfrancis.com Fri Sep 24 07:26:35 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Water bath antigen retrieval Message-ID: I do the very same procedure Tom listed. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK 74136 japoteete@saintfrancis.com > -----Original Message----- > From: Tom McNemar [SMTP:TMcNemar@lmhealth.org] > Sent: Friday, September 24, 2004 6:36 AM > To: 'Paula Pierce'; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Water bath antigen retrieval > > Wondering if this is your standard procedure for all antibodies? I only > steam for 20 minutes with a 20 minute cool for all retrieval. Place them > in > buffer for 5 minutes then directly on the Autostainer. They work > beautifully. > > Just curious, > > Tom Mc Nemar HT(ASCP) > Histology Supervisor > Licking Memorial Hospital > Newark, Ohio 43055 > > > > -----Original Message----- > From: Paula Pierce [mailto:contact@excaliburpathology.com] > Sent: Thursday, September 23, 2004 2:05 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Water bath antigen retrieval > > > I use a Black and Decker rice steamer for antigen retrieval. About $30. > Fill > the bottom with distilled water. Place the plastic basket in the steamer. > Place a container with your slides and citrate buffer in the basket so the > lid will fit. Turn the steamer on for 1 hour, then let cool for 25 > minutes. > Rinse in distilled water slowly to finish cooling. Proceed with your IHC > stain. > > > > > _______________________________________________ > Histonet mailing list > Histonet@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 contact <@t> excaliburpathology.com Fri Sep 24 08:46:50 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Water bath antigen retrieval Message-ID: <20040924134650.95757.qmail@web50308.mail.yahoo.com> Tom, yes this is my standard method. If yours work with only 20 minutes of heating that is great to know. I may try cutting my time to 30 minutes. Paula From kosmicdog <@t> hotmail.com Fri Sep 24 09:19:25 2004 From: kosmicdog <@t> hotmail.com (jason m) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] IHC - weak signals Message-ID: Does anyone have advice for amplifying weak signals from IHC? Is there a detection system better than DAB which can make the signal more noticable? _________________________________________________________________ Designer Mail isn't just fun to send, it's fun to receive. Use special stationery, fonts and colors. http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines Start enjoying all the benefits of MSN® Premium right now and get the first two months FREE*. From mprice26 <@t> juno.com Fri Sep 24 09:59:32 2004 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] RE: MACH 4 Universal Polymer Message-ID: <20040924.075935.5684.773976@webmail18.nyc.untd.com> Dear histonetters, I am interested in trying the MACH 4 Polymer from BIOCARE. Is anyone out there using in there lab and if so could you give me some feed back on how it works for you. Thank you in advance. Marsha Price ________________________________________________________________ Get your name as your email address. Includes spam protection, 1GB storage, no ads and more Only $1.99/ month - visit http://www.mysite.com/name today! From dobbin <@t> upei.ca Fri Sep 24 10:02:05 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] IHC - weak signals In-Reply-To: Message-ID: <41540CBC.32258.BE6D63@localhost> Hi Jason, Post more details about your procedure. The chromagen step is only one of many steps that can contribute to signal strength, and at that, not the most likely place to start. Tell the group what you are staining for, with frozens or paraffin sections, how long and at what temp the primary is applied, with what detection system, if any retreival methods are being employed, etc. We need the whole story before we can offer any help. Cheers! Greg From: "jason m" To: histonet@pathology.swmed.edu BCC to: Date sent: Fri, 24 Sep 2004 07:19:25 -0700 Copies to: Subject: [Histonet] IHC - weak signals > Does anyone have advice for amplifying weak signals from IHC? Is there a > detection system better than DAB which can make the signal more noticable? > > _________________________________________________________________ > Designer Mail isn't just fun to send, it's fun to receive. Use special > stationery, fonts and colors. > http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines > Start enjoying all the benefits of MSN? Premium right now and get the > first two months FREE*. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 vazquezr <@t> ohsu.edu Fri Sep 24 10:25:39 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] IHC - weak signals Message-ID: Hello, I use the Ted Pella microwave for tissue processing. Does anyone have a good procedure for this. It is a hit and miss sometimes. I don't use JFC anymore, but instead I use reagent alcohol. The instructions I was given was if I process the same day, I let the tissue sit in non-buffered formalin for ? hour then process 15-20 min (which had to be cut down to 2-3 min), then rinse 95% for 2 min, then into 100% absol 15-20min(down to 2-3 min) 65 C, JFC sol the same absolute but at 70 c. I changed the JFC sol for reagent alc still no change. The tissue does work out most of the time, but it's those times that it doesn't, makes it frustrating. Then into paraffin at 83 C the same time down to 2-3 min. My tissue would shrink and be crunchie. I tried the saline fixative yesterday with extra tissue. It shrank 50%, but was not crunchie. Do I have my paraffin to hot? thanks for your input.... Robyn Vazquez >>> "Greg Dobbin" 9/24/2004 12:02:05 PM >>> Hi Jason, Post more details about your procedure. The chromagen step is only one of many steps that can contribute to signal strength, and at that, not the most likely place to start. Tell the group what you are staining for, with frozens or paraffin sections, how long and at what temp the primary is applied, with what detection system, if any retreival methods are being employed, etc. We need the whole story before we can offer any help. Cheers! Greg From: "jason m" To: histonet@pathology.swmed.edu BCC to: Date sent: Fri, 24 Sep 2004 07:19:25 -0700 Copies to: Subject: [Histonet] IHC - weak signals > Does anyone have advice for amplifying weak signals from IHC? Is there a > detection system better than DAB which can make the signal more noticable? > > _________________________________________________________________ > Designer Mail isn't just fun to send, it's fun to receive. Use special > stationery, fonts and colors. > http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines > Start enjoying all the benefits of MSN? Premium right now and get the > first two months FREE*. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 jbrielma <@t> yahoo.com Fri Sep 24 10:43:49 2004 From: jbrielma <@t> yahoo.com (Jennifer Brielmaier) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Cresyl violet vs. cresyl violet acetate Message-ID: <20040924154349.12846.qmail@web61306.mail.yahoo.com> Hello everyone, I am a first-year graduate student in a biopsychology program and am enrolled in a basic histology course this semester. Today in class we are going to learn how to make cresyl violet stain solutions. Our instructor has informed us that we have a "mystery" bottle in the lab; it is not known whether it is cresyl violet or cresyl violet acetate. Can anyone tell me whether there is a simple test that can be performed that will tell us which solution is in the bottle? Thanks very much. Jennifer --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From la.sebree <@t> hosp.wisc.edu Fri Sep 24 11:09:08 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] RE: MACH 4 Universal Polymer Message-ID: Hi Marsha, I have limited experience with Biocare's polymer system but what I've used it for has worked well. I'm currently using it on a research project using renal biopsies. It makes all that nasty biotin just disappear! 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: mprice26@juno.com [mailto:mprice26@juno.com] Sent: Friday, September 24, 2004 10:00 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: MACH 4 Universal Polymer Dear histonetters, I am interested in trying the MACH 4 Polymer from BIOCARE. Is anyone out there using in there lab and if so could you give me some feed back on how it works for you. Thank you in advance. Marsha Price ________________________________________________________________ Get your name as your email address. Includes spam protection, 1GB storage, no ads and more Only $1.99/ month - visit http://www.mysite.com/name today! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From galinadeyneko <@t> yahoo.com Fri Sep 24 11:39:53 2004 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] (no subject) Message-ID: <20040924163953.61087.qmail@web14521.mail.yahoo.com> Hi Marcel, It looks like you wash out cells on previous steps, pipette the reagents into well very slowly, do not pipette on the membrane directly, only underneath of membrane into well using small holes. Do not use tissue processor, process only manually. I believe you use xylene substitute. I use Shandon HistoSolve # 9990505 from Thermo Shandon. Last step in processing is paraffin. Use intermediate step 1 part of xylene sub+ 1 part of liquid paraffin. In the oven 58- 60 C i have 3 containers 1:1 Xylene sub: paraffin- 30 min Paraffin 1- 1 hour Paraffin 2- 1 hour. I take out insert (not membrane) from well and incubate in consecutive order in containers.After last paraffin I take out insert, carefully poor off the excess of paraffin and leave thin layer of paraffin on both side of membrane and let solidify at room T. Layers prevent cells on membrane. Using warm surgical blade cut out membrane covered by paraffin from insert, cut on 2 or 3 pieces by scissors and embed on routine way, it means you embed a piece of paraffin in paraffin. Galina. --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From rkrug <@t> sial.com Fri Sep 24 13:18:37 2004 From: rkrug <@t> sial.com (Robert Krug) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Long term storage of tissue Message-ID: Peggy: Thanks for sharing your expertise. I would also like to thank the other people who have responded to my query. I have a copy of Bancroft and Gamble. I also have the 2nd and 4th Editions of Bancroft and Stevens. The 2nd Edition has a nice section on preparing Museum Quality tissues. The chapter implies the tissues seemed to be used only for viewing and the tissues are processed and stored in various Kaiserling solutions. No mention is ever made to the tissues being utilized for anything except viewing. What I am really trying to locate is a study or a textbook which states at what intervals the formalin should be changed out - or does it ever need to be changed. For every reference I find documenting the formation of formic acid, I find another piece of information which simply states tissues may be stored in formalin forever. From the time I first started working in Histology, I was always told that tissues stored in formalin had to have the formalin changed on a yearly basis. I can't remember that we ever changed formalin solutions. There were no OSHA requirements for formaldehyde monitoring at that time. Changing the solutions or disposing of formalin fixed tissues was always a dreaded experience in the days before OSHA reclassified formaldehyde. The bottles of formaldehyde simply advised the user to "seek fresh air if you feel light headed." If we needed to archive tissue, the tissue was always fixed in formalin and placed in 70% ethanol before storing. Why? Because that's the way the lab had always handled tissue. I do have some experience with spirochette controls which were fixed in formalin for around 1 week and then stored in 70% alcohol. This tissue was eventually embedded into paraffin blocks, but the tissue was probably 5 years old before the last of the tissue was embedded. The staining with the Steiner Steiner procedure remained unchanged from the time the tissue was first processed to the time the last wet tissue was section and processed. When I was in the Army, the entire block of tissue from the visceration was stored in a large pan and submersed in formalin. The bodies went to the funeral directors without the tissues being returned. The volume of 10% NBF was never really sufficient to properly fix the large volume of tissue retained. Eventually the formalin evaporated and fungus would sometimes be noted on the tissues. If the reaction were unreversible, why the formation of fungus? Although the center of the tissues were unquestionably poorly fixed, it seems the tissue surface should have been fixed. By that time the fungal growth was noted, the cases had been signed out and the tissues were incincerated. I seriously doubt if many labs today would consider handling human tissues the way we did in the 70s. Many thanks Bob Krug St John, Missouri 09/23/2004 06:06 PM To: "Robert Krug" cc: Subject: Re: [Histonet] Long term storage of tissue To build on what van der Loos and Pierce stated - Yes, formic acid is created when formaldehyde reacts with oxygen. Long term storage in formic acid, even low levels, will cause destruction of proteins in the tissue. Yes, additional cross-links are formed, which can interfere with staining, by blocking epitopes or distorting the proteins so they are not stainable. >From personal experience. One year, we got a great case of lung TB from one autopsy, and a wonderful kidney with amyloid from another autopsy. We grossed a lot of each for controls, and stored the rest in formalin. About 7 years later, we had run out of the blocks of controls, and went back to the stored wet stock, and grossed a bunch more blocks. Neither would now stain positive for the AFB or the amyloid. They could still be "seen" on the H&E, but the Kinyoun would no longer stain the mycobacterium, nor would the Congo red stain the amyloid. No matter how we altered the stain (increase time, heat, less differentiation, etc.). Now, I don't know if the proteins in the TB cell wall and the amyloid proteins were ruined by acid destruction or excess formalin cross-links or by protein distortion. It really didn't matter the root reason. Basic fact - the tissue would no longer stain. Lost were two cases of great control material. As for a source - How about "Theory and Practice of Histological Techniques" 5th ed., John D. Bancroft and Marilyn Gamble, 2002, Churchill-Livingstone. In the chapter on "fixation", under the topic "duration": "Long fixation in these aldehydes (fixatives) is known to severely inhibit enzyme activity and immunological reactions. This has been attributed to limitations in substrate diffusion and cross-link formation between protein molecules." Most histotechnique textbooks discuss the formation of formic acid with long term storage of tissue in formalin. Hope this helps. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Robert Krug" To: Sent: Wednesday, September 22, 2004 2:57 PM Subject: [Histonet] Long term storage of tissue > I have a question concerning the long term storage of tissue which is not > addressed in any of the histology textbooks I have on hand. After many > hours of searching, I was unable to find an answer through a websearch. > > Can tissue be preserved in 10% neutral buffered formalin for decades > without changing the 10% NBF (provided the container is sealed and does > not leak)? > > A former coworker of mine once worked at the CDC and she stated the CDC > used 70% alcohol for the long term storage of tissue. A review of the > Internet shows that the Australian Museum Fish site states they have fixed > in "formaldehyde and then stored in alcohol" for the last 80 years. This > site claims to have thousands of specimens on hand. Another site I > found recommended storage in formalin as a way to keep the original color > of the tissue and recommended against long term storage of tissue in > alcohol for various reasons. > > The unused 10% neutral buffered formalin solution does decompose over time > and is typically assigned an expiration period. Once the tissue has been > fixed in 10% neutral buffered formalin, is the reaction stable, meaning > the bonds are stable and no decomposition will occur? I was always told > the formalin solution degrades with time and may form formic acid and > other by products. Given enough time the remaining formalin in solution > might form formic acid, drop out of solution as paraformaldehyde or > escape as formaldehyde gas. Another coworker feels that once fixation > has occurred, the fixation is permanent and there is no need to change the > solution. I realize the fixation bonds can be broken through washing in > running water and with select chemicals. But given a sealed container, > will the bonds remain permanent and the tissue free of decomposition or > fungal growth? > > Any jounal articles or textbook references you have to support your view > point would be greatly appreciated. > > Many thanks > > Bob Krug > St John, Missouri > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From flemons <@t> bhset.org Fri Sep 24 13:53:43 2004 From: flemons <@t> bhset.org (Fran Lemons) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Reflections of a first timer Message-ID: This is going to be the name of my article I am going to write for our state newsletter............discussing the wonders of my first time at the national convention!! Just wanted to say, it was a total success in my book and I enjoyed meeting everyone! Oh yeah, & I learned some stuff, too. Much appreciation to everyone who assisted this babe in the wood! Fran Walker From TMcNemar <@t> lmhealth.org Fri Sep 24 14:15:05 2004 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] CAP surveys for IHC..... Message-ID: <90092A4ED388D7119575006008F7112049CBE0@NT_EXCHANGE> Hello all, I was wondering which IHC surveys you folks subscribe to. For years we have done the Immunhistochemistry survey. This year I also did the MTTB (PM1, PM2, PM3, & PM4). I wasn't going to buy the MTTB one again because it is the same antibodies that I did this year (CD20, EGFR, CD117, ER). Apparantly these are the only antibodies covered by the MTTB. I understand the reason behind doing these surveys and that they serve different purposes. Since I've already done the 4 MTTB and since each of those antibodies are at one time or another used in the Immunohistochemistry survey, I really don't see the need to buy the MTTB survey again if it is going to be the same antibodies. So that's my question..... Do you folks do both? One or the other? Or something else? Thanks in advance. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 From miller <@t> coho.net Fri Sep 24 14:36:50 2004 From: miller <@t> coho.net (Diane G. Miller) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Looking for Fran Lee Adams Message-ID: <000a01c4a26d$d75b0ce0$0200a8c0@desktop> Hello All, I'm trying to contact Fran Lee Adams. Fran if you're out there would you please contact me? If anyone knows Fran's contact information, would you please let her know I'm looking for her? As always thanks to everyone for your help. Thanks Diane Diane G. Miller HT(ASCP), QIHC 503-784-6444 miller@coho.net From Robert.Lott <@t> bhsala.com Fri Sep 24 14:37:00 2004 From: Robert.Lott <@t> bhsala.com (Lott, Robert) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] IHC for Fibrinogen Message-ID: <35B6C610DD1DD311B1FA0008C791400407E14683@gobexchm3.bhsala.com> Is anyone out in histonet land doing IHC for fibrinogen on paraffin sections...if so, what antibody concentration do you use ... what retrieval method is used? Thanks, Robert Robert L. Lott, HTL(ASCP) Manager, Anatomic Pathology Baptist Health System 800 Montclair Road Birmingham, AL 35213 205-592-5388 phone 205-592-5646 fax robert.lott@bhsala.com Confidentiality Notice: The information contained in this email message is privileged and confidential information and intended only for the use of the individual or entity named in the address. If you are not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this information is strictly prohibited. If you received this information in error, please notify the sender and delete this information from your computer and retain no copies of any of this information. From Cathy.Stevens <@t> HealthONEcares.com Fri Sep 24 14:57:30 2004 From: Cathy.Stevens <@t> HealthONEcares.com (Stevens Cathy) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] histotech position Message-ID: We are looking for a registered HT to do basic histology. We don't perform Immunos or any other specials except routine special stains. It is a full time position. We are located in Aurora, Colorado. If you are interested please respond to this emial. Thank you. Cathy Stevens H.T.(ASCP) BS Pathology Coordinator The Medical Center of Aurora P-303-695-2636 F-303-873-5660 cathy.stevens@healthonecares.com 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 recieved 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 recieve 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. From Wanda.Smith <@t> HCAhealthcare.com Fri Sep 24 15:18:46 2004 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Global Billing Message-ID: <8784A3EE34199644AF60DBE517F5B0A604A4185A@louex04.lou.medcity.net> Dear Netters, We are in the process of returning to global billing, where the Pathology group bills the patients for the technical and professional fees and pays the hospital for the technical services. Previously, the Pathologist paid the hospital "per block" and per frozen section block, but they did not pay for any special stains, decal charges or multiple slides for levels on biopsy specimens, etc. We would like to be fair, without cutting our own throats! Can anyone that does global billing share the price and billing structure with me? Any help on this would be greatly appreciated! Thanks, Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > From kutaisi13 <@t> hotmail.com Sat Sep 25 04:25:13 2004 From: kutaisi13 <@t> hotmail.com (david moses) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] NeuN Message-ID: Dear histonetters I'm trying to make NeuN work on 25 micron mounted brain sections, but no luck so far. Has anyone got suggestions/protocols? Thanks in advance David Moses Howard Florey institute _________________________________________________________________ STOP MORE SPAM with the new MSN 8 and get 2 months FREE* http://join.msn.com/?page=features/junkmail From settembr <@t> umdnj.edu Sat Sep 25 06:32:05 2004 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] CAP surveys for IHC..... Message-ID: We do the CAP survey only for IHC Dana Settembre University Hospital - UMDNJ Newark, NJ USA >>> Tom McNemar 9/24/2004 3:15:05 PM >>> Hello all, I was wondering which IHC surveys you folks subscribe to. For years we have done the Immunhistochemistry survey. This year I also did the MTTB (PM1, PM2, PM3, & PM4). I wasn't going to buy the MTTB one again because it is the same antibodies that I did this year (CD20, EGFR, CD117, ER). Apparantly these are the only antibodies covered by the MTTB. I understand the reason behind doing these surveys and that they serve different purposes. Since I've already done the 4 MTTB and since each of those antibodies are at one time or another used in the Immunohistochemistry survey, I really don't see the need to buy the MTTB survey again if it is going to be the same antibodies. So that's my question..... Do you folks do both? One or the other? Or something else? Thanks in advance. 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 cuttingedgehistology <@t> yahoo.com Sat Sep 25 12:45:40 2004 From: cuttingedgehistology <@t> yahoo.com (Brandon Stokes) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] CLIA Inspection..... Message-ID: <20040925174540.22640.qmail@web41407.mail.yahoo.com> Hello everyone:) I am having a CLIA inspection for my IHC and DIF procedures late on Monday. Anyone have any advice on anything to watch out for and make sure not to miss as far as what details are commonly overlooked? I would appreciate any advice anyone might have. Brandon Stokes, HT(ASCP) Lab Manager Cutting Edge Histology Services, LLC Tigard, OR --------------------------------- Do you Yahoo!? Yahoo! Mail Address AutoComplete - You start. We finish. From lpwenk <@t> sbcglobal.net Sat Sep 25 13:41:20 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Cresyl violet vs. cresyl violet acetate References: <20040924154349.12846.qmail@web61306.mail.yahoo.com> Message-ID: <004201c4a32f$41337be0$933ad445@domainnotset.invalid> I have a question, concerning your "mystery" bottle - Do you mean that the teacher has no idea what it is (other than it might be a cresyl-family dye), and would like some idea/help on how to figure out what it is? Or, is this an assignment, where each student is supposed to find out the chemical nature of each of the dyes, and thus be able to chemically prove which dye it is. Where you are getting a grade for this. Be honest with us. If the teacher actually doesn't know, I think the Histonet community would be willing to help. If this is your homework assignment, I think that the Histonet community would be willing to refer you to a text book where you could look up the answers yourself, but that we would not be willing to do your homework for you. Sorry if I'm sounding a little edgy. I'm an instructor in histotechnology. All of my assignments are designed to teach students about histotechnology. But there is more to our field than just science. Some of my assignments are "team" assignments, where my students can pull together their knowledge and abilities to work on the assignment together. Part of what I'm assess is the ability of each student to contribute to a team. Some of my other assignments are "solo", where the person can use book, journals, internet, etc., but they cannot get the answers from their classmates or other techs. Part of what I'm assessing is the person's ability to find the answer on their own, not be told it. I need to know whether a person can problem-solve and troubleshoot on their own. Either way, there are text books and web pages out there with the information you need about your dyes. I'm sitting here with one of the books in my lab right now, which has the information. So - if your teacher needs the help, I am willing to help and quote from the pages. If you are supposed to find the answer, I'll let you know the name of the book. And you can look it up for yourself (and hopefully learn about this book, and learn about other dyes while leafing through this book). So, again, be honest, and let me know. Peggy A. Wenk, BA, BS, HTL(ASCP)SLS Program Director School of Histologic Technicians School of Histotechnologist William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Jennifer Brielmaier" To: Sent: Friday, September 24, 2004 11:43 AM Subject: [Histonet] Cresyl violet vs. cresyl violet acetate > Hello everyone, > > I am a first-year graduate student in a biopsychology program and am enrolled in a basic histology course this semester. Today in class we are going to learn how to make cresyl violet stain solutions. Our instructor has informed us that we have a "mystery" bottle in the lab; it is not known whether it is cresyl violet or cresyl violet acetate. Can anyone tell me whether there is a simple test that can be performed that will tell us which solution is in the bottle? Thanks very much. > > Jennifer > > > --------------------------------- > Do you Yahoo!? > vote.yahoo.com - Register online to vote today! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DRMOSES <@t> peoplepc.com Sat Sep 25 20:16:30 2004 From: DRMOSES <@t> peoplepc.com (David Moses) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Histology review class for HT exam Message-ID: <001401c4a366$74af3e50$5ac495ce@turbo> There is a 3 day course offered in Boston, for people due to take their HT exam. I was wondering if anyone has taken this course by Theresa Schuldt? And was it helpful? Please send your comments directly to my email. Thanks. From RFail <@t> Charleston.net Sun Sep 26 05:43:34 2004 From: RFail <@t> Charleston.net (Rena) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Mart-1 Message-ID: <000501c4a3b5$acdba3a0$8a11a6a5@renad4yk9b8abe> For those of you using Mart-1, which clone do you prefer. Have you experienced any problems with this antibody? Thank you Rena Fail From RFail <@t> Charleston.net Sun Sep 26 05:40:29 2004 From: RFail <@t> Charleston.net (Rena) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Polyclonals vs Monoclonals Message-ID: <000001c4a3b5$3eb93630$8a11a6a5@renad4yk9b8abe> I know that monoclonals are more specific than polyclonals. The question is which is preferred. Do you have both and use the polyclonal as a screening antibody, then use the monoclonal? Do you prefer monoclonals or polyclonals and why? Thank you Rena Fail From jkiernan <@t> uwo.ca Sun Sep 26 12:43:33 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Polyclonals vs Monoclonals References: <000001c4a3b5$3eb93630$8a11a6a5@renad4yk9b8abe> Message-ID: <4156FFC5.63CB12C2@uwo.ca> A monoclonal antibody (MAB) binds to one tiny bit of the antigen molecule, called an epitope, which typically is a particular sequence of a few amino acids in a particular conformation (shape). If the epitope is masked (for example, by other nearby protein molecules) a MAB cannot bind to it. If the same epitope exists also in a different protein - not the antigen to which the antibody was raised - then the MAB will bind to the wrong antigen. (That is not to say false-positives are abundant in immunohistochemistry with MABs.) A polyclonal antiserum contains numerous antibodies, each with specificity for a different epitope of the antigen, so there is a better chance of some antibody molecules binding to the antigen that you are seeking to stain. The positive immunostaining results from summation of the different antibodies. In an antiserum there will also be antibodies that can bind to epitopes in proteins other than the antigen of interest. These can give rise to false-positive or background staining. There are techniques for reducing unwanted immunostaining: notably absorption of the antiserum with a tissue powder that does not contain the antigen, and a simple chromatographic procedure called affinity purification. The leaflet with a commercially supplied antiserum should tell you if these procedures have been applied to the product. In short, one cannot in general prefer monoclonals or polyclonals. Either may be better for a certain job. It's important to do the classical control procedures when doing imunohistochemistry, aspecially with a combination of antibody and tissue that you haven't worked with before. John Kiernan London, Canada. ---------------------------------------------------- Rena wrote: > > I know that monoclonals are more specific than polyclonals. The question > is which is preferred. Do you have both and use the polyclonal as a > screening antibody, then use the monoclonal? Do you prefer monoclonals > or polyclonals and why? > > Thank you > Rena Fail > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Sun Sep 26 15:41:51 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Leptospira PCR Message-ID: Anyone doing PCR for leptospirosis? If so, what types of specimens do you test? 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 mariatere <@t> infovia.com.ar Sun Sep 26 18:58:00 2004 From: mariatere <@t> infovia.com.ar (Teresa Dominguez) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Iron Stain References: Message-ID: <001801c4a424$aa76b220$887c46c8@DFI> Hi jessica, I don't performe mouse tissues, but I found some techniques in the Manual of Histologic Staining Methods, or you can visit some sites like http://members.pgonline.com/%7Ebryand/StainsFile/stain/stainmenu.htm http://www.nottingham.ac.uk/pathology/default.html Good luck! Ht. Maria T Dominguez H.R.R.G. Pathology Service, Rio Grande, Tierra del Fuego, Argentina. ----- Original Message ----- From: "Jessica Butler" To: Sent: Thursday, September 23, 2004 10:36 AM Subject: [Histonet] Iron Stain > Hi- > I work at Emory University and we are currently investigating sickle cell > disease. I need a protocol for staining mouse lung tissue for iron. The > protocol and/or stain can be for paraffin fixed or frozen sections. Thanks- > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Sun Sep 26 23:58:34 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Iron Stain References: <001801c4a424$aa76b220$887c46c8@DFI> Message-ID: <41579DFA.2AFB3121@uwo.ca> Iron! The choice of staining method depends on the required sensitivity of the histochemical detection of iron. Try the Perls method or one of its common variants (in any techniques book published in the last 50+ years). It may well be adequate, and it is easy to do. The Prussian blue reaction product can fade in some mounting media, so take photographs less than one year after preparing the slides. At Emory University, with its excellent libraries, you should have no difficulty finding some books that explain the Perls method. Click on http://web.library.emory.edu/services/ressvcs/catalogs.html and you will soon be on your way to the right building, floor and shelf. If old Perls's (= Perls') technique is not sensitive enough for your needs, there are ways to enhance it, and also to convert the Prussian blue to a permanently visible coloured product. There are also staining methods for iron that use chemistry other than Prussian/Turnbull's blue reactions. For more advanced iron histochemistry, see a modern histochemistry book (= Pearse Vol.2, 1985 or something more recent) and follow up the references to the original papers. For research work you should always go to the source even if a textbook provides apparently simple instructions. Please send a message to Histonet when you have found a method that shows iron in sickle-cell anaemia lungs. Others may well benefit from your reported trials and errors. -- ------------------------------- 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 _______________________________ Jessica Butler > I work at Emory University and we are currently investigating sickle cell > disease. I need a protocol for staining mouse lung tissue for iron. The > protocol and/or stain can be for paraffin fixed or frozen sections. > Thanks- From juan.gutierrez <@t> christushealth.org Mon Sep 27 08:24:07 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Cresyl violet vs. cresyl violet acetate Message-ID: Isn't looking in the histonet almost the same as if looking it up in a book? At least an effort is being made to look in the right place. What do you all think? I think the fact that Jennifer was able to find us means that she put some effort into it. Shouldn't we reward her? I'll keep my answer on hold until we can find a consensus. Good luck Jennifer. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 -----Original Message----- From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] Sent: Saturday, September 25, 2004 1:41 PM To: Jennifer Brielmaier; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Cresyl violet vs. cresyl violet acetate I have a question, concerning your "mystery" bottle - Do you mean that the teacher has no idea what it is (other than it might be a cresyl-family dye), and would like some idea/help on how to figure out what it is? Or, is this an assignment, where each student is supposed to find out the chemical nature of each of the dyes, and thus be able to chemically prove which dye it is. Where you are getting a grade for this. Be honest with us. If the teacher actually doesn't know, I think the Histonet community would be willing to help. If this is your homework assignment, I think that the Histonet community would be willing to refer you to a text book where you could look up the answers yourself, but that we would not be willing to do your homework for you. Sorry if I'm sounding a little edgy. I'm an instructor in histotechnology. All of my assignments are designed to teach students about histotechnology. But there is more to our field than just science. Some of my assignments are "team" assignments, where my students can pull together their knowledge and abilities to work on the assignment together. Part of what I'm assess is the ability of each student to contribute to a team. Some of my other assignments are "solo", where the person can use book, journals, internet, etc., but they cannot get the answers from their classmates or other techs. Part of what I'm assessing is the person's ability to find the answer on their own, not be told it. I need to know whether a person can problem-solve and troubleshoot on their own. Either way, there are text books and web pages out there with the information you need about your dyes. I'm sitting here with one of the books in my lab right now, which has the information. So - if your teacher needs the help, I am willing to help and quote from the pages. If you are supposed to find the answer, I'll let you know the name of the book. And you can look it up for yourself (and hopefully learn about this book, and learn about other dyes while leafing through this book). So, again, be honest, and let me know. Peggy A. Wenk, BA, BS, HTL(ASCP)SLS Program Director School of Histologic Technicians School of Histotechnologist William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Jennifer Brielmaier" To: Sent: Friday, September 24, 2004 11:43 AM Subject: [Histonet] Cresyl violet vs. cresyl violet acetate > Hello everyone, > > I am a first-year graduate student in a biopsychology program and am enrolled in a basic histology course this semester. Today in class we are going to learn how to make cresyl violet stain solutions. Our instructor has informed us that we have a "mystery" bottle in the lab; it is not known whether it is cresyl violet or cresyl violet acetate. Can anyone tell me whether there is a simple test that can be performed that will tell us which solution is in the bottle? Thanks very much. > > Jennifer > > > --------------------------------- > Do you Yahoo!? > vote.yahoo.com - Register online to vote today! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Mon Sep 27 13:36:21 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Cresyl violet vs. cresyl violet acetate In-Reply-To: References: Message-ID: <41585DA5.8040302@umdnj.edu> Weighing in on this topic ....... GUTIERREZ, JUAN wrote: >Isn't looking in the histonet almost the same as if looking it up in a book? > If she knew which book to use and where to find it she would have the information by now. > At least an effort is being made to look in the right place. >What do you all think? > She need to be able to use the library, after all she is a graduate student! >I think the fact that Jennifer was able to find us means that she put some effort into it. > I don't know how much effort she put into finding us, maybe she just used google? > Shouldn't we reward her? > The knowledge and knowing where to look for it should be the reward. There are a lot of resources that, due to their age, will probably never be digitalized and available on the internet. Students need to be able to use the library. Histonet does not work too well on the weekends or when the server is down. My $0.02 worth. Geoff >I'll keep my answer on hold until we can find a consensus. Good luck Jennifer. > >Juan C. Gutierrez, HT(ASCP) >Histology Laboratory Supervisor >(210)704-2533 > > >-----Original Message----- >From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] >Sent: Saturday, September 25, 2004 1:41 PM >To: Jennifer Brielmaier; histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Cresyl violet vs. cresyl violet acetate > >I have a question, concerning your "mystery" bottle - > >Do you mean that the teacher has no idea what it is (other than it might be >a cresyl-family dye), and would like some idea/help on how to figure out >what it is? > >Or, is this an assignment, where each student is supposed to find out the >chemical nature of each of the dyes, and thus be able to chemically prove >which dye it is. Where you are getting a grade for this. > >Be honest with us. > >If the teacher actually doesn't know, I think the Histonet community would >be willing to help. > >If this is your homework assignment, I think that the Histonet community >would be willing to refer you to a text book where you could look up the >answers yourself, but that we would not be willing to do your homework for >you. > >Sorry if I'm sounding a little edgy. I'm an instructor in histotechnology. >All of my assignments are designed to teach students about histotechnology. >But there is more to our field than just science. Some of my assignments are >"team" assignments, where my students can pull together their knowledge and >abilities to work on the assignment together. Part of what I'm assess is the >ability of each student to contribute to a team. Some of my other >assignments are "solo", where the person can use book, journals, internet, >etc., but they cannot get the answers from their classmates or other techs. >Part of what I'm assessing is the person's ability to find the answer on >their own, not be told it. I need to know whether a person can problem-solve >and troubleshoot on their own. > >Either way, there are text books and web pages out there with the >information you need about your dyes. I'm sitting here with one of the books >in my lab right now, which has the information. > >So - if your teacher needs the help, I am willing to help and quote from the >pages. > >If you are supposed to find the answer, I'll let you know the name of the >book. And you can look it up for yourself (and hopefully learn about this >book, and learn about other dyes while leafing through this book). > >So, again, be honest, and let me know. > >Peggy A. Wenk, BA, BS, HTL(ASCP)SLS >Program Director >School of Histologic Technicians >School of Histotechnologist >William Beaumont Hospital >Royal Oak, MI 48073 > >----- Original Message ----- >From: "Jennifer Brielmaier" >To: >Sent: Friday, September 24, 2004 11:43 AM >Subject: [Histonet] Cresyl violet vs. cresyl violet acetate > > > > >>Hello everyone, >> >>I am a first-year graduate student in a biopsychology program and am >> >> >enrolled in a basic histology course this semester. Today in class we are >going to learn how to make cresyl violet stain solutions. Our instructor has >informed us that we have a "mystery" bottle in the lab; it is not known >whether it is cresyl violet or cresyl violet acetate. Can anyone tell me >whether there is a simple test that can be performed that will tell us which >solution is in the bottle? Thanks very much. > > >>Jennifer >> >> >>--------------------------------- >>Do you Yahoo!? >>vote.yahoo.com - Register online to vote today! >>_______________________________________________ >>Histonet mailing list >>Histonet@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 > > > -- -- ********************************************** 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 kwuny <@t> email.cs.nsw.gov.au Sun Sep 26 19:54:20 2004 From: kwuny <@t> email.cs.nsw.gov.au (Young Kwun) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Collagen IV subtypes:alpha 3 - alpha 6 Message-ID: <200409271050329.SM05496@crgcsls814> Dear all, Does anyone know where I can get antibodies to subtypes of all collagen IV chains (alpha 1?alpha 6)? Are they only available for immunofluorescence detection? SantaCruz has some antibodies for collagen IV alpha 1, alpha 2 and alpha 3 isoforms. I also know that ID Labs in Canada has a clone specific to alpha 1 and alpha 2. Thank you. Young Young Kwun 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 _______________________________________________ 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 peoshel <@t> wisc.edu Sun Sep 26 12:53:33 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Polyclonals vs Monoclonals In-Reply-To: <4156FFC5.63CB12C2@uwo.ca> References: <000001c4a3b5$3eb93630$8a11a6a5@renad4yk9b8abe> <4156FFC5.63CB12C2@uwo.ca> Message-ID: If I may, I'd like to add a bit to John's comment: affinity. Affinity is essentially the strength with which an antibody binds to an epitope. With monoclonal antibodies, the Ab will only have the one affinity, and if it is not strong, then there will be a weak single, even if the monoclonal Ab is strongly specific for the epitope in question. If the affinity is strong, then that's great. Just not likely. Polyclonal Abs will not only have varying specificities, they have varying affinities, so there is a better chance of getting an Ab that not only is specific for the epitopes sought, but also has a high affinity (or affinities) for the epitope(s). Phil >A monoclonal antibody (MAB) binds to one tiny bit of the >antigen molecule, called an epitope, which typically >is a particular sequence of a few amino acids in >a particular conformation (shape). If the epitope >is masked (for example, by other nearby protein >molecules) a MAB cannot bind to it. If the same >epitope exists also in a different protein - not the >antigen to which the antibody was raised - then >the MAB will bind to the wrong antigen. (That is not >to say false-positives are abundant in immunohistochemistry >with MABs.) > >A polyclonal antiserum contains numerous antibodies, >each with specificity for a different epitope of >the antigen, so there is a better chance of some >antibody molecules binding to the antigen that you >are seeking to stain. The positive immunostaining >results from summation of the different antibodies. > >In an antiserum there will also be antibodies that >can bind to epitopes in proteins other than the >antigen of interest. These can give rise to >false-positive or background staining. There are >techniques for reducing unwanted immunostaining: >notably absorption of the antiserum with a tissue >powder that does not contain the antigen, and a >simple chromatographic procedure called affinity >purification. The leaflet with a commercially >supplied antiserum should tell you if these >procedures have been applied to the product. > >In short, one cannot in general prefer monoclonals >or polyclonals. Either may be better for a >certain job. It's important to do the classical >control procedures when doing imunohistochemistry, >aspecially with a combination of antibody and >tissue that you haven't worked with before. > > John Kiernan > London, Canada. >---------------------------------------------------- >Rena wrote: >> >> I know that monoclonals are more specific than polyclonals. The question >> is which is preferred. Do you have both and use the polyclonal as a >> screening antibody, then use the monoclonal? Do you prefer monoclonals >> or polyclonals and why? >> >> Thank you >> Rena Fail >> _______________________________________________ >> Histonet mailing list >> Histonet@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, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From jhabecke <@t> seattlecca.org Mon Sep 27 11:11:52 2004 From: jhabecke <@t> seattlecca.org (Randolph-Habecker, Julie) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Processing lung tissue after pulmonary function tests Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EACA6@wala01.seattlecca.org> Histonetters, I am working with an investigator who is studying lung function after lung transplantation. We would like to look at FFPE lung tissue after the subject has had lung function tests which involve fluorescent plastic beads of varying size - the largest being 15 microns. I am concerned that the normal processing could melt the beads and cause artifact. Someone in another lab suggested that we use Histoclear II rather than Xylene. Does anyone have experience in processing tissue like this? Please give me some information on how to handle this tissue. Thanks! Julie Julie Randolph-Habecker, Ph.D. Experimental Histopathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave. N, G1-300 PO Box 19023 Seattle, WA 98109-1024 Tel: (206) 288-1187 FAX: (206) 288-1345 jhabecke@fhcrc.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 dobbin <@t> upei.ca Mon Sep 27 11:51:37 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Processing lung tissue after pulmonary function tests In-Reply-To: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EACA6@wala01.seattlecca.org> Message-ID: <41581AE8.8485.121771D@localhost> Hi Julie, It seems to me that a solvent is a solvent is a solvent! The plastic beads (polystyrene I suppose?) will dissolve in which ever solvent is used to process the tissue. Is frozen sectioning out of the question for some reason? Greg From: "Randolph-Habecker, Julie" To: "'histonet@lists.utsouthwestern.edu'" Date sent: Mon, 27 Sep 2004 09:11:52 -0700 Subject: [Histonet] Processing lung tissue after pulmonary function tests > Histonetters, > > I am working with an investigator who is studying lung function after lung > transplantation. We would like to look at FFPE lung tissue after the subject > has had lung function tests which involve fluorescent plastic beads of > varying size - the largest being 15 microns. I am concerned that the normal > processing could melt the beads and cause artifact. Someone in another lab > suggested that we use Histoclear II rather than Xylene. > > Does anyone have experience in processing tissue like this? Please give me > some information on how to handle this tissue. > > Thanks! > > Julie > > Julie Randolph-Habecker, Ph.D. > Experimental Histopathology Shared Resources > Fred Hutchinson Cancer Research Center > 1100 Fairview Ave. N, G1-300 > PO Box 19023 > Seattle, WA 98109-1024 > Tel: (206) 288-1187 > FAX: (206) 288-1345 > jhabecke@fhcrc.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. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 contact <@t> excaliburpathology.com Mon Sep 27 12:32:29 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Processing lung tissue .... Message-ID: <20040927173229.58115.qmail@web50306.mail.yahoo.com> I use X-S Xylene substitute to process my eye tissue. It does not dissolve the artificial intraocular lenses placed in eyes after cataract surgery nor plastic buckles around post trauma blind painful eyes that are removed. However, after sectioning xylene used in the staining procedure will dissolve these out. So you may also need to change your staining setup. And, you may also need to change the mounting media used for coverslipping as X-S does not mix well with media such as Permount. You can obtain X-S Xylene substitute from StatLab Medical Products in Texas. Their website is www.statlab.com. Paula Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From carl.hobbs <@t> kcl.ac.uk Mon Sep 27 13:44:21 2004 From: carl.hobbs <@t> kcl.ac.uk (carl hobbs) Date: Fri Sep 16 15:24:04 2005 Subject: [Histonet] Re: Polyclonals vs Monoclonals Message-ID: The definitive answer from an expert( John Kiernan). My NB on your query would be to not use the terms ?less/more specific?. It is either specific or not. There are no degrees; however , there are degrees of selectivity. A pedantic point, but I was , after all, taught by the Great Harry Cook and, if he was allowed, would?ve thumped me every time I discussed degrees of specificity in my essays lol. With great respect to him. --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.762 / Virus Database: 510 - Release Date: 13/09/2004 From Laurie.Pereira <@t> sdcounty.ca.gov Mon Sep 27 13:59:57 2004 From: Laurie.Pereira <@t> sdcounty.ca.gov (Pereira, Laurie ) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] salaries Message-ID: Hi guys! I am planning on moving to Florida. Just wondering what histotechnicians get paid in Florida? Any information would be wonderful :) Laurie From BlazekL <@t> childrensdayton.org Mon Sep 27 14:07:53 2004 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] salaries Message-ID: Brave woman, moving to Florida! Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 1 Children's Plaza Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org From jhabecke <@t> seattlecca.org Mon Sep 27 14:16:34 2004 From: jhabecke <@t> seattlecca.org (Randolph-Habecker, Julie) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Processing lung tissue after pulmonary function te sts Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EACAD@wala01.seattlecca.org> Greg, I agree with the solvent issue. We have already frozen part of the tissue for frozen sections as well. Thanks, Julie -----Original Message----- From: Greg Dobbin [mailto:dobbin@upei.ca] Sent: Monday, September 27, 2004 6:52 AM To: Randolph-Habecker, Julie Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Processing lung tissue after pulmonary function tests Hi Julie, It seems to me that a solvent is a solvent is a solvent! The plastic beads (polystyrene I suppose?) will dissolve in which ever solvent is used to process the tissue. Is frozen sectioning out of the question for some reason? Greg From: "Randolph-Habecker, Julie" To: "'histonet@lists.utsouthwestern.edu'" Date sent: Mon, 27 Sep 2004 09:11:52 -0700 Subject: [Histonet] Processing lung tissue after pulmonary function tests > Histonetters, > > I am working with an investigator who is studying lung function after lung > transplantation. We would like to look at FFPE lung tissue after the subject > has had lung function tests which involve fluorescent plastic beads of > varying size - the largest being 15 microns. I am concerned that the normal > processing could melt the beads and cause artifact. Someone in another lab > suggested that we use Histoclear II rather than Xylene. > > Does anyone have experience in processing tissue like this? Please give me > some information on how to handle this tissue. > > Thanks! > > Julie > > Julie Randolph-Habecker, Ph.D. > Experimental Histopathology Shared Resources > Fred Hutchinson Cancer Research Center > 1100 Fairview Ave. N, G1-300 > PO Box 19023 > Seattle, WA 98109-1024 > Tel: (206) 288-1187 > FAX: (206) 288-1345 > jhabecke@fhcrc.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. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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. 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 Luis.Chiriboga <@t> med.nyu.edu Mon Sep 27 14:57:21 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] salaries In-Reply-To: Message-ID: after this year whatever they are paying, it's not enough!!!!! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Linda Blazek Sent: Monday, September 27, 2004 3:08 PM To: histonet@lists.utsouthwestern.edu; Laurie.Pereira@sdcounty.ca.gov Subject: Re: [Histonet] salaries Brave woman, moving to Florida! Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 1 Children's Plaza Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Mon Sep 27 15:24:53 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Cresyl violet vs. cresyl violet acetate Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4078@fh2k093.fhmis.net> I find the most important skill is knowing your resources and where to look for the answer. -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Monday, September 27, 2004 2:36 PM To: GUTIERREZ, JUAN Cc: histonet@lists.utsouthwestern.edu; lpwenk@sbcglobal.net Subject: Re: [Histonet] Cresyl violet vs. cresyl violet acetate Weighing in on this topic ....... GUTIERREZ, JUAN wrote: >Isn't looking in the histonet almost the same as if looking it up in a book? > If she knew which book to use and where to find it she would have the information by now. > At least an effort is being made to look in the right place. >What do you all think? > She need to be able to use the library, after all she is a graduate student! >I think the fact that Jennifer was able to find us means that she put some effort into it. > I don't know how much effort she put into finding us, maybe she just used google? > Shouldn't we reward her? > The knowledge and knowing where to look for it should be the reward. There are a lot of resources that, due to their age, will probably never be digitalized and available on the internet. Students need to be able to use the library. Histonet does not work too well on the weekends or when the server is down. My $0.02 worth. Geoff >I'll keep my answer on hold until we can find a consensus. Good luck Jennifer. > >Juan C. Gutierrez, HT(ASCP) >Histology Laboratory Supervisor >(210)704-2533 > > >-----Original Message----- >From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] >Sent: Saturday, September 25, 2004 1:41 PM >To: Jennifer Brielmaier; histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Cresyl violet vs. cresyl violet acetate > >I have a question, concerning your "mystery" bottle - > >Do you mean that the teacher has no idea what it is (other than it might be >a cresyl-family dye), and would like some idea/help on how to figure out >what it is? > >Or, is this an assignment, where each student is supposed to find out the >chemical nature of each of the dyes, and thus be able to chemically prove >which dye it is. Where you are getting a grade for this. > >Be honest with us. > >If the teacher actually doesn't know, I think the Histonet community would >be willing to help. > >If this is your homework assignment, I think that the Histonet community >would be willing to refer you to a text book where you could look up the >answers yourself, but that we would not be willing to do your homework for >you. > >Sorry if I'm sounding a little edgy. I'm an instructor in histotechnology. >All of my assignments are designed to teach students about histotechnology. >But there is more to our field than just science. Some of my assignments are >"team" assignments, where my students can pull together their knowledge and >abilities to work on the assignment together. Part of what I'm assess is the >ability of each student to contribute to a team. Some of my other >assignments are "solo", where the person can use book, journals, internet, >etc., but they cannot get the answers from their classmates or other techs. >Part of what I'm assessing is the person's ability to find the answer on >their own, not be told it. I need to know whether a person can problem-solve >and troubleshoot on their own. > >Either way, there are text books and web pages out there with the >information you need about your dyes. I'm sitting here with one of the books >in my lab right now, which has the information. > >So - if your teacher needs the help, I am willing to help and quote from the >pages. > >If you are supposed to find the answer, I'll let you know the name of the >book. And you can look it up for yourself (and hopefully learn about this >book, and learn about other dyes while leafing through this book). > >So, again, be honest, and let me know. > >Peggy A. Wenk, BA, BS, HTL(ASCP)SLS >Program Director >School of Histologic Technicians >School of Histotechnologist >William Beaumont Hospital >Royal Oak, MI 48073 > >----- Original Message ----- >From: "Jennifer Brielmaier" >To: >Sent: Friday, September 24, 2004 11:43 AM >Subject: [Histonet] Cresyl violet vs. cresyl violet acetate > > > > >>Hello everyone, >> >>I am a first-year graduate student in a biopsychology program and am >> >> >enrolled in a basic histology course this semester. Today in class we are >going to learn how to make cresyl violet stain solutions. Our instructor has >informed us that we have a "mystery" bottle in the lab; it is not known >whether it is cresyl violet or cresyl violet acetate. Can anyone tell me >whether there is a simple test that can be performed that will tell us which >solution is in the bottle? Thanks very much. > > >>Jennifer >> >> >>--------------------------------- >>Do you Yahoo!? >>vote.yahoo.com - Register online to vote today! >>_______________________________________________ >>Histonet mailing list >>Histonet@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 > > > -- -- ********************************************** 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 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 jestrong <@t> earthlink.net Mon Sep 27 15:25:07 2004 From: jestrong <@t> earthlink.net (Jes Strong - Source Medical Products) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] salaries In-Reply-To: Message-ID: I hope Florida is still there. My heartfelt support for all of you who have had to suffer the seemingly endless challenges you have had to face the past couple of months. I know it may be of little condolence, but we all care and pray for you. Jes Strong Source Medical Products Representatives for Statlab, DuraEdge, Bladex, Diagnostic BioSystems, RA Lamb and other Quality Pathology Manufacturers (847) 323-8373 (Cell) (847) 735-9965 (Office) (800) 442-3573 x250 (Voice Mail) (508) 861-1575 (Fax) jestrong@earthlink.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Linda Blazek Sent: Monday, September 27, 2004 2:08 PM To: histonet@lists.utsouthwestern.edu; Laurie.Pereira@sdcounty.ca.gov Subject: Re: [Histonet] salaries Brave woman, moving to Florida! Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 1 Children's Plaza Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Mon Sep 27 15:37:56 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] salaries Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4079@fh2k093.fhmis.net> Thank you! This hurricane business: no electric, no water, long lines at the gas station, I guess you just learn the drill. It reminds me of camping. The workplace actually becomes an oasis during these times!! Food, water, lights, air-conditioning....aahhhhhh- -Original Message----- From: Jes Strong - Source Medical Products [mailto:jestrong@earthlink.net] Sent: Monday, September 27, 2004 4:25 PM To: Linda Blazek; histonet@lists.utsouthwestern.edu; Laurie.Pereira@sdcounty.ca.gov Subject: RE: [Histonet] salaries I hope Florida is still there. My heartfelt support for all of you who have had to suffer the seemingly endless challenges you have had to face the past couple of months. I know it may be of little condolence, but we all care and pray for you. Jes Strong Source Medical Products Representatives for Statlab, DuraEdge, Bladex, Diagnostic BioSystems, RA Lamb and other Quality Pathology Manufacturers (847) 323-8373 (Cell) (847) 735-9965 (Office) (800) 442-3573 x250 (Voice Mail) (508) 861-1575 (Fax) jestrong@earthlink.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Linda Blazek Sent: Monday, September 27, 2004 2:08 PM To: histonet@lists.utsouthwestern.edu; Laurie.Pereira@sdcounty.ca.gov Subject: Re: [Histonet] salaries Brave woman, moving to Florida! Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 1 Children's Plaza Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.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 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 jkiernan <@t> uwo.ca Mon Sep 27 15:39:31 2004 From: jkiernan <@t> uwo.ca (J. A. Kiernan) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Cresyl violet vs. cresyl violet acetate References: Message-ID: <41587A83.D5424CDA@uwo.ca> "GUTIERREZ, JUAN" wrote: > > Isn't looking in the histonet almost the same as if looking it up in a book? No! -- ------------------------------- 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 _______________________________ > At least an effort is being made to look in the right place. > What do you all think? > I think the fact that Jennifer was able to find us means that she put some effort into it. Shouldn't we reward her? > I'll keep my answer on hold until we can find a consensus. Good luck Jennifer. > > Juan C. Gutierrez, HT(ASCP) > Histology Laboratory Supervisor > (210)704-2533 > > -----Original Message----- > From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] > Sent: Saturday, September 25, 2004 1:41 PM > To: Jennifer Brielmaier; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Cresyl violet vs. cresyl violet acetate > > I have a question, concerning your "mystery" bottle - > > Do you mean that the teacher has no idea what it is (other than it might be > a cresyl-family dye), and would like some idea/help on how to figure out > what it is? > > Or, is this an assignment, where each student is supposed to find out the > chemical nature of each of the dyes, and thus be able to chemically prove > which dye it is. Where you are getting a grade for this. > > Be honest with us. > > If the teacher actually doesn't know, I think the Histonet community would > be willing to help. > > If this is your homework assignment, I think that the Histonet community > would be willing to refer you to a text book where you could look up the > answers yourself, but that we would not be willing to do your homework for > you. > > Sorry if I'm sounding a little edgy. I'm an instructor in histotechnology. > All of my assignments are designed to teach students about histotechnology. > But there is more to our field than just science. Some of my assignments are > "team" assignments, where my students can pull together their knowledge and > abilities to work on the assignment together. Part of what I'm assess is the > ability of each student to contribute to a team. Some of my other > assignments are "solo", where the person can use book, journals, internet, > etc., but they cannot get the answers from their classmates or other techs. > Part of what I'm assessing is the person's ability to find the answer on > their own, not be told it. I need to know whether a person can problem-solve > and troubleshoot on their own. > > Either way, there are text books and web pages out there with the > information you need about your dyes. I'm sitting here with one of the books > in my lab right now, which has the information. > > So - if your teacher needs the help, I am willing to help and quote from the > pages. > > If you are supposed to find the answer, I'll let you know the name of the > book. And you can look it up for yourself (and hopefully learn about this > book, and learn about other dyes while leafing through this book). > > So, again, be honest, and let me know. > > Peggy A. Wenk, BA, BS, HTL(ASCP)SLS > Program Director > School of Histologic Technicians > School of Histotechnologist > William Beaumont Hospital > Royal Oak, MI 48073 > > ----- Original Message ----- > From: "Jennifer Brielmaier" > To: > Sent: Friday, September 24, 2004 11:43 AM > Subject: [Histonet] Cresyl violet vs. cresyl violet acetate > > > Hello everyone, > > > > I am a first-year graduate student in a biopsychology program and am > enrolled in a basic histology course this semester. Today in class we are > going to learn how to make cresyl violet stain solutions. Our instructor has > informed us that we have a "mystery" bottle in the lab; it is not known > whether it is cresyl violet or cresyl violet acetate. Can anyone tell me > whether there is a simple test that can be performed that will tell us which > solution is in the bottle? Thanks very much. > > > > Jennifer > > > > > > --------------------------------- > > Do you Yahoo!? > > vote.yahoo.com - Register online to vote today! > > _______________________________________________ > > Histonet mailing list > > Histonet@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 ljb <@t> medicine.wisc.edu Mon Sep 27 15:44:13 2004 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] salaries Message-ID: No matter what they offer, you better negotiate for a heft hazard pay differential! good luck >>> "Pereira, Laurie " 09/27/04 01:59PM >>> Hi guys! I am planning on moving to Florida. Just wondering what histotechnicians get paid in Florida? Any information would be wonderful :) Laurie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kzhong888 <@t> yahoo.com Mon Sep 27 17:34:08 2004 From: kzhong888 <@t> yahoo.com (dfs dsaf) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] IEC Daemon or CTD harris Message-ID: <20040927223408.55378.qmail@web41609.mail.yahoo.com> Dear histonetters: our office is trying to expand our mohs lab, and we need a couple of IEC cryostats. i would like to get a smaller Daemon or CTD-harris cryostats (ice cream carts) if possible. we are also interested in the larger Minotome models. If you know of anyone ready to sell them please contact me ASAP. i would be happy to arrange shipping or any other logistical issues. please email me at kzhong888@yahoo.com or call 626-432-5032. thank you again histonetters Kirk Zhong --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - 100MB free storage! From sagitalcuts <@t> yahoo.com Mon Sep 27 17:55:06 2004 From: sagitalcuts <@t> yahoo.com (Yolanda Maldonado) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Vimentin on mouse tissue Message-ID: <20040927225506.66447.qmail@web53507.mail.yahoo.com> Hi everyone: Does anyone out there has any experience with IHC stain for Vimentin on mouse tissue? I have tried the V9 and 3B4 clone from different companies as well as the ARK kit and other polymers...Nothing seems to work for me!! I have tried EDTA. CB, Reveal, LAB and enzymes as AR and I have also run human and other species along with my tissue to make sure my staining is working. Everything stains but the mice. Any suggestions? __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From jbrielma <@t> yahoo.com Mon Sep 27 19:35:56 2004 From: jbrielma <@t> yahoo.com (Jennifer Brielmaier) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] to clear things up... Message-ID: <20040928003556.63881.qmail@web61302.mail.yahoo.com> To everyone who has been debating over my earlier post, I would like to clear things up here. The question of the "mystery bottle" was a question asked by our teacher on an informal basis as something for us to think about until we could discuss it in the next week's class. It was NOT a graded assignment nor did it pertain to some kind of activity that I was supposed to perform on my own. I looked for the information in my textbook and did not find anything. I looked up a some information elsewhere on the Internet and was unable to find an answer. Our instructor has encouraged us to use the Histonet listserv as a resource. In fact, she frequently reads the postings and when she saw my post, she replied to me saying she was pleased I'd used Histonet. In the class meeting following the one where the question had been asked, we figured out the contents of the mystery bottle as a class, which is what we had planned to do all along. Ironically, the simple test she was thinking of to use was checking the pH, which is what I'd had in mind all along! Again, this was a simple, informal question asked of us, so I saw no problem with posting it to Histonet and neither did my instructor. Thanks to everyone who gave me helpful answers. I am a hard worker and I certainly know how to use a library. I feel a bit insulted by those who have implied otherwise... Jennifer --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - Send 10MB messages! From jbrielma <@t> yahoo.com Mon Sep 27 19:40:43 2004 From: jbrielma <@t> yahoo.com (Jennifer Brielmaier) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] one more thing... Message-ID: <20040928004043.45451.qmail@web61309.mail.yahoo.com> I forgot to mention in my last post that our instructor had no idea what was in the bottle either, so it was a surprise for all of us when we did find out! --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - 100MB free storage! From Ianbernard <@t> netzero.com Mon Sep 27 20:38:26 2004 From: Ianbernard <@t> netzero.com (ian bernard) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] salaries In-Reply-To: Message-ID: Wait till after hurricane season. Let me know what you found out. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pereira, Laurie Sent: Monday, September 27, 2004 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] salaries Hi guys! I am planning on moving to Florida. Just wondering what histotechnicians get paid in Florida? Any information would be wonderful :) Laurie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Mon Sep 27 22:09:33 2004 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Can't get back on Message-ID: <00ca01c4a508$9489b270$f76fce44@yourxhtr8hvc4p> Hello again, ya know, one day I'll be computer literate. I can't get back on the Histonet. My emails to the Histonet get returned. How am I to get flamed if I can't respond. Heeeeeeelllllllllllllpppppppppppp please. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: Joe Nocito To: histonet Sent: Tuesday, September 14, 2004 8:56 PM Subject: Flame me Okay, for those of you who are interested or care, on Sunday, I will be wearing a navy blue T-shirt with a red and white target and a star in the center. I'll be the guy with the beard and wearing a flapjacket. Hope everyone has a safe trip to Toronto, watch out for Ivan and LET THE FLAMING BEGIN. See ya Sunday Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX From sebres <@t> comcast.net Mon Sep 27 22:46:37 2004 From: sebres <@t> comcast.net (sebres) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] to clear things up... (was: Cresyl violet vs. cresyl violet acetate) References: <20040928003556.63881.qmail@web61302.mail.yahoo.com> Message-ID: <001c01c4a50d$c1a8ce80$0900a8c0@RESLAPTOP> Sounds like it's time for me to jump in & smooth any ruffled feathers! I'm the infamous teacher who posed the "mystery bottle" question. Our class was the grateful recipient of a generous donation of old cresyl violet stain of uncertain identity from Anatech last year. I thought it might liven up the task of learning how to follow recipes to make up stains to work through the challenge together of solving this "mystery", which, as Jennifer says, we determined by pH'g the solution & comparing to known cresyl violet acetate. On the one hand, I am trying to encourage my students to participate in Histonet, and am pleased that at least one student has tuned in. On the other, I must (& will) convey more clearly the need to exhaust available information (e.g. comparing the 2 protocols) before resorting to Histonet! Jennifer, to clarify a bit, many on Histonet are educators as well as histologists, & I sense that they felt some conflict between desire to aid a novice & concern that this could compromise what they astutely recognized as a classroom exercise (as Jennifer said, this was an informal challenge, not a graded exercise), no offense to your industriousness intended. (She really is a very dedicated student!) I was very touched that several of you took great pains to help, and in addition, to thoughtfully consider the ulterior motive (i.e. mine) behind the question. Anyway, please accept my thanks in addition to Jennifer's, and I would humbly hope that the consolation prize for the commotion we inadvertantly stirred up might include these useful tidbits: 1) in case anyone else out there runs into a similar dilemma, pH is indeed a simple solution 2) some thought provoking various opinions on the question of what questions it is fair to post to Histonet & what alternative sources of information should be consulted first 3) a reminder to me to emphasize to my students the netiquette of sticking with the original subject line on a listserve 4) it has been a fun exercise to make learning how to make stains more interesting, that perhaps other instructors could utilize 5) finally, I hope Peggy Wenk will forgive me for quoting some fascinating history that she sent me when I confessed that I still wasn't clear myself on how cresyl FAST violet compares to the other variants: "I respond with a little bit more info - Cresyl echt violet (echt = fast, as in dye-fast, so the English translation is cresyl fast violet) was made only in Germany. No one else had the "recipe". Germany had a monopoly on most dyes in the early 1900's. After WW I or WW II, I don't remember), Germany no longer had the ability to make this dye (chemical plants were destroyed). And no one knew the formulation for CEV. A substitute was eventually made that was almost, but not quite, CEV. This was/is cresyl violet acetate. However, most companies continued to call it CEV. It's only recently that I'm seeing more companies calling it CVA. Which is confusing everyone one, since the books and procedures are still calling for CEV. Peggy" Finally, I have to commend all of you folks on Histonet for contributing to such a generous, collegial and valuable resource! I sincerely hope we haven't abused the privilege of taking part in this community, or offended anyone! with my deep gratitude, Susan Bachus, George Mason University ----- Original Message ----- From: "Jennifer Brielmaier" To: Sent: Monday, September 27, 2004 8:35 PM Subject: [Histonet] to clear things up... > To everyone who has been debating over my earlier post, > > I would like to clear things up here. The question of the "mystery bottle" was a question asked by our teacher on an informal basis as something for us to think about until we could discuss it in the next week's class. It was NOT a graded assignment nor did it pertain to some kind of activity that I was supposed to perform on my own. I looked for the information in my textbook and did not find anything. I looked up a some information elsewhere on the Internet and was unable to find an answer. Our instructor has encouraged us to use the Histonet listserv as a resource. In fact, she frequently reads the postings and when she saw my post, she replied to me saying she was pleased I'd used Histonet. > > In the class meeting following the one where the question had been asked, we figured out the contents of the mystery bottle as a class, which is what we had planned to do all along. Ironically, the simple test she was thinking of to use was checking the pH, which is what I'd had in mind all along! Again, this was a simple, informal question asked of us, so I saw no problem with posting it to Histonet and neither did my instructor. > > Thanks to everyone who gave me helpful answers. I am a hard worker and I certainly know how to use a library. I feel a bit insulted by those who have implied otherwise... > > Jennifer > > > --------------------------------- > Do you Yahoo!? > New and Improved Yahoo! Mail - Send 10MB messages! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Tue Sep 28 05:30:51 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Cresyl violet vs. cresyl violet acetate Message-ID: You're right. It's actually better. With our busy schedules a lot of times we don't have the time to sit down and leaf through a bunch of books looking for solutions to our problems. It only takes a couple of minutes to write an e-mail and after completing some of our other tasks, it's nice to come back and find the answer waiting for you on the net. My two cents. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 -----Original Message----- From: J. A. Kiernan [mailto:jkiernan@uwo.ca] Sent: Monday, September 27, 2004 3:40 PM To: GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu Cc: lpwenk@sbcglobal.net Subject: Re: [Histonet] Cresyl violet vs. cresyl violet acetate "GUTIERREZ, JUAN" wrote: > > Isn't looking in the histonet almost the same as if looking it up in a book? No! -- ------------------------------- 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 _______________________________ > At least an effort is being made to look in the right place. > What do you all think? > I think the fact that Jennifer was able to find us means that she put some effort into it. Shouldn't we reward her? > I'll keep my answer on hold until we can find a consensus. Good luck Jennifer. > > Juan C. Gutierrez, HT(ASCP) > Histology Laboratory Supervisor > (210)704-2533 > > -----Original Message----- > From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] > Sent: Saturday, September 25, 2004 1:41 PM > To: Jennifer Brielmaier; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Cresyl violet vs. cresyl violet acetate > > I have a question, concerning your "mystery" bottle - > > Do you mean that the teacher has no idea what it is (other than it might be > a cresyl-family dye), and would like some idea/help on how to figure out > what it is? > > Or, is this an assignment, where each student is supposed to find out the > chemical nature of each of the dyes, and thus be able to chemically prove > which dye it is. Where you are getting a grade for this. > > Be honest with us. > > If the teacher actually doesn't know, I think the Histonet community would > be willing to help. > > If this is your homework assignment, I think that the Histonet community > would be willing to refer you to a text book where you could look up the > answers yourself, but that we would not be willing to do your homework for > you. > > Sorry if I'm sounding a little edgy. I'm an instructor in histotechnology. > All of my assignments are designed to teach students about histotechnology. > But there is more to our field than just science. Some of my assignments are > "team" assignments, where my students can pull together their knowledge and > abilities to work on the assignment together. Part of what I'm assess is the > ability of each student to contribute to a team. Some of my other > assignments are "solo", where the person can use book, journals, internet, > etc., but they cannot get the answers from their classmates or other techs. > Part of what I'm assessing is the person's ability to find the answer on > their own, not be told it. I need to know whether a person can problem-solve > and troubleshoot on their own. > > Either way, there are text books and web pages out there with the > information you need about your dyes. I'm sitting here with one of the books > in my lab right now, which has the information. > > So - if your teacher needs the help, I am willing to help and quote from the > pages. > > If you are supposed to find the answer, I'll let you know the name of the > book. And you can look it up for yourself (and hopefully learn about this > book, and learn about other dyes while leafing through this book). > > So, again, be honest, and let me know. > > Peggy A. Wenk, BA, BS, HTL(ASCP)SLS > Program Director > School of Histologic Technicians > School of Histotechnologist > William Beaumont Hospital > Royal Oak, MI 48073 > > ----- Original Message ----- > From: "Jennifer Brielmaier" > To: > Sent: Friday, September 24, 2004 11:43 AM > Subject: [Histonet] Cresyl violet vs. cresyl violet acetate > > > Hello everyone, > > > > I am a first-year graduate student in a biopsychology program and am > enrolled in a basic histology course this semester. Today in class we are > going to learn how to make cresyl violet stain solutions. Our instructor has > informed us that we have a "mystery" bottle in the lab; it is not known > whether it is cresyl violet or cresyl violet acetate. Can anyone tell me > whether there is a simple test that can be performed that will tell us which > solution is in the bottle? Thanks very much. > > > > Jennifer > > > > > > --------------------------------- > > Do you Yahoo!? > > vote.yahoo.com - Register online to vote today! > > _______________________________________________ > > Histonet mailing list > > Histonet@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 anaflorenti <@t> yahoo.com Tue Sep 28 05:52:52 2004 From: anaflorenti <@t> yahoo.com (Ana Florentin) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Antibody for Rat kidney Fibroblasts? Message-ID: <20040928105252.10576.qmail@web52604.mail.yahoo.com> Hi everybody! Does anybody know about an antibody for rats kidney fibroblasts?! I would appreciate any suggestion. Thanks a lot, anne _______________________________ Do you Yahoo!? Declare Yourself - Register online to vote today! http://vote.yahoo.com From Catherine.Goeden <@t> med.va.gov Tue Sep 28 09:12:43 2004 From: Catherine.Goeden <@t> med.va.gov (Goeden, Catherine) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Trimming blocks Message-ID: <597FCC5997E8504EAA6814E6C9ED52298FAD67@VHASUXEXC1> We are using the new Thermo hotplate in my lab and I love it!!! Sorry for the late reply! -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Monday, August 30, 2004 10:37 AM To: Rebecca Barnhart; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Trimming blocks an old solid scalpel blade. For a nice setup, look at Thermo Electron's (Shandon) hot plate, tilted for this purpose- I would love one. At 08:38 AM 8/30/2004, you wrote: >Just curious what everyone else is using to trim the excess paraffin off >the blocks? Thanks for the input. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From aep10 <@t> cornell.edu Tue Sep 28 09:17:26 2004 From: aep10 <@t> cornell.edu (Anna Elisse Pflaster) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] (no subject) Message-ID: <1763.128.253.96.73.1096381046.squirrel@128.253.96.73> Hello, I am going to start processing mouse embryos for in situ and immunohistochemistry, and since I am a novice, I have a few very basic questions that I am hoping to find answers to! First, I was wondering if it is absolutely necessary to embed embryos in paraffin for cryosectioning. I usually work with brain tissue from adult mice, and I never have to embed in paraffin -- I always freeze in OCT. In this case, I will be sectioning embryos at ED14.5 and ED17.5. Also, I was wondering about a simple fixation procedure for 'older' embryos. Is it feasible for fixation to involve only fixing cryosections in 4% paraformaldehyde? Lastly, if my goal is to look at expression patterns in the embryonic brain, what would be the best approach/angle to sectioning? Specifically, I am interested in having a good view of the ventricular zone and dentate gyrus. I would greatly appreciate any and all advice on these matters! Thank you in advance! Anna Beaudin, M.Sc. Division of Nutritional Sciences Cornell University Ithaca NY 14850 From katri <@t> cogeco.ca Tue Sep 28 10:03:18 2004 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Cresyl violet vs. cresyl violet acetate References: Message-ID: <002701c4a56c$49f9b890$6a9a9618@Katri> I love Histonet, but it is not better or worse than books and other resources. It is different! It can create discussions, give you a right direction for your problem solving and encourage further search for information from books and other publications. It certainly has it's place in this fast moving technology of ours, but you have to have an open mind, when interpreting the responses. I enjoyed all five days of NSH Convention in Toronto and got reminded once again, that nothing is written in stone and advances in technology can overthrow the old "proven" standards. Always keep an open mind... Katri Katri Tuomala St.Joseph's Healthcare Hamilton, Ontario, Canada ----- Original Message ----- From: "GUTIERREZ, JUAN" To: "J. A. Kiernan" ; Cc: Sent: Tuesday, September 28, 2004 6:30 AM Subject: RE: [Histonet] Cresyl violet vs. cresyl violet acetate You're right. It's actually better. With our busy schedules a lot of times we don't have the time to sit down and leaf through a bunch of books looking for solutions to our problems. It only takes a couple of minutes to write an e-mail and after completing some of our other tasks, it's nice to come back and find the answer waiting for you on the net. My two cents. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 -----Original Message----- From: J. A. Kiernan [mailto:jkiernan@uwo.ca] Sent: Monday, September 27, 2004 3:40 PM To: GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu Cc: lpwenk@sbcglobal.net Subject: Re: [Histonet] Cresyl violet vs. cresyl violet acetate "GUTIERREZ, JUAN" wrote: > > Isn't looking in the histonet almost the same as if looking it up in a book? No! -- ------------------------------- 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 _______________________________ > At least an effort is being made to look in the right place. > What do you all think? > I think the fact that Jennifer was able to find us means that she put some effort into it. Shouldn't we reward her? > I'll keep my answer on hold until we can find a consensus. Good luck Jennifer. > > Juan C. Gutierrez, HT(ASCP) > Histology Laboratory Supervisor > (210)704-2533 > > -----Original Message----- > From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] > Sent: Saturday, September 25, 2004 1:41 PM > To: Jennifer Brielmaier; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Cresyl violet vs. cresyl violet acetate > > I have a question, concerning your "mystery" bottle - > > Do you mean that the teacher has no idea what it is (other than it might be > a cresyl-family dye), and would like some idea/help on how to figure out > what it is? > > Or, is this an assignment, where each student is supposed to find out the > chemical nature of each of the dyes, and thus be able to chemically prove > which dye it is. Where you are getting a grade for this. > > Be honest with us. > > If the teacher actually doesn't know, I think the Histonet community would > be willing to help. > > If this is your homework assignment, I think that the Histonet community > would be willing to refer you to a text book where you could look up the > answers yourself, but that we would not be willing to do your homework for > you. > > Sorry if I'm sounding a little edgy. I'm an instructor in histotechnology. > All of my assignments are designed to teach students about histotechnology. > But there is more to our field than just science. Some of my assignments are > "team" assignments, where my students can pull together their knowledge and > abilities to work on the assignment together. Part of what I'm assess is the > ability of each student to contribute to a team. Some of my other > assignments are "solo", where the person can use book, journals, internet, > etc., but they cannot get the answers from their classmates or other techs. > Part of what I'm assessing is the person's ability to find the answer on > their own, not be told it. I need to know whether a person can problem-solve > and troubleshoot on their own. > > Either way, there are text books and web pages out there with the > information you need about your dyes. I'm sitting here with one of the books > in my lab right now, which has the information. > > So - if your teacher needs the help, I am willing to help and quote from the > pages. > > If you are supposed to find the answer, I'll let you know the name of the > book. And you can look it up for yourself (and hopefully learn about this > book, and learn about other dyes while leafing through this book). > > So, again, be honest, and let me know. > > Peggy A. Wenk, BA, BS, HTL(ASCP)SLS > Program Director > School of Histologic Technicians > School of Histotechnologist > William Beaumont Hospital > Royal Oak, MI 48073 > > ----- Original Message ----- > From: "Jennifer Brielmaier" > To: > Sent: Friday, September 24, 2004 11:43 AM > Subject: [Histonet] Cresyl violet vs. cresyl violet acetate > > > Hello everyone, > > > > I am a first-year graduate student in a biopsychology program and am > enrolled in a basic histology course this semester. Today in class we are > going to learn how to make cresyl violet stain solutions. Our instructor has > informed us that we have a "mystery" bottle in the lab; it is not known > whether it is cresyl violet or cresyl violet acetate. Can anyone tell me > whether there is a simple test that can be performed that will tell us which > solution is in the bottle? Thanks very much. > > > > Jennifer > > > > > > --------------------------------- > > Do you Yahoo!? > > vote.yahoo.com - Register online to vote today! > > _______________________________________________ > > Histonet mailing list > > Histonet@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 CMCCOLLOUGH <@t> dnr.state.md.us Tue Sep 28 10:06:10 2004 From: CMCCOLLOUGH <@t> dnr.state.md.us (McCollough, Carol) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Malmberg's fixative Message-ID: Greetings Histonetters: I have been asked for a recipe for Malmberg's fixative (1:1 ammonium picrate and glycerol). I cannot find a method for the preparation of ammonium picrate. Does anyone have a recipe or a commercial source for this nasty? Thanks very much. Regards - Carol ********************** Carol B. McCollough, HT/HTL(ASCP) Diagnostics & Histology Laboratory Manager Oyster Disease Research Project Fisheries Service Maryland Department of Natural Resources Cooperative Oxford Laboratory 904 S. Morris Street Oxford, Maryland 21654 410-226-5193 From Barry.R.Rittman <@t> uth.tmc.edu Tue Sep 28 10:34:43 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Cresyl violet vs. cresyl violet acetate Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0F13947@UTHEVS3.mail.uthouston.edu> Histonet is a great resource and like most resources should be used appropriately. The discussions that we have recently seen reflect a general tendency in our society for us to be fed complete pieces of information rather than to provide a pathway for us to be able to follow a logical sequence of events. I teach histology and it is obvious that most students expect to be spoon fed information that has been examined, selected and summarized by us for them. We are now in a situation where the impetus seems to be to pass examinations rather than to learn material. (can you tell I have spent 3 days marking examinations?) I believe that part of this tendency is the rush to complete tasks and with so many demands on our time everything seems to be prioritized. The other problem is that with so many resources it is often difficult to know the most appropriate source for information. I see many people turning to Histonet because they have been thrown into a situation without adequate training or complete instruction as to the final outcome that is desired. I feel great sympathy for these individuals. This is akin in some ways to asking individuals who have never painted to produce a full length self portrait using acrylics. I strongly believe that Histonet shines in providing these individuals with guidance as to where to look. I do not think that giving them a complete final answer is as beneficial as providing them with general directions and appropriate sites to look. It is always very tempting to provide a final answer but I feel that this does not necessarily help the learning process of such individuals. To this end I suggest that in response to this type of questions that general direction be given and that emphasis be placed on directly contacting those who have submitted answers until the problem is solved. Once solved to then supply the final answer to Histonet so that we can all benefit. Barry 713-500-4134 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Katri Tuomala Sent: Tuesday, September 28, 2004 10:03 AM To: GUTIERREZ, JUAN; J. A. Kiernan; histonet@lists.utsouthwestern.edu Cc: lpwenk@sbcglobal.net Subject: Re: [Histonet] Cresyl violet vs. cresyl violet acetate I love Histonet, but it is not better or worse than books and other resources. It is different! It can create discussions, give you a right direction for your problem solving and encourage further search for information from books and other publications. It certainly has it's place in this fast moving technology of ours, but you have to have an open mind, when interpreting the responses.e I enjoyed all five days of NSH Convention in Toronto and got reminded once again, that nothing is written in stone and advances in technology can overthrow the old "proven" standards. Always keep an open mind... Katri Katri Tuomala St.Joseph's Healthcare Hamilton, Ontario, Canada ----- Original Message ----- From: "GUTIERREZ, JUAN" To: "J. A. Kiernan" ; Cc: Sent: Tuesday, September 28, 2004 6:30 AM Subject: RE: [Histonet] Cresyl violet vs. cresyl violet acetate You're right. It's actually better. With our busy schedules a lot of times we don't have the time to sit down and leaf through a bunch of books looking for solutions to our problems. It only takes a couple of minutes to write an e-mail and after completing some of our other tasks, it's nice to come back and find the answer waiting for you on the net. My two cents. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 -----Original Message----- From: J. A. Kiernan [mailto:jkiernan@uwo.ca] Sent: Monday, September 27, 2004 3:40 PM To: GUTIERREZ, JUAN; histonet@lists.utsouthwestern.edu Cc: lpwenk@sbcglobal.net Subject: Re: [Histonet] Cresyl violet vs. cresyl violet acetate "GUTIERREZ, JUAN" wrote: > > Isn't looking in the histonet almost the same as if looking it up in a book? No! -- ------------------------------- 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 _______________________________ > At least an effort is being made to look in the right place. > What do you all think? > I think the fact that Jennifer was able to find us means that she put some effort into it. Shouldn't we reward her? > I'll keep my answer on hold until we can find a consensus. Good luck Jennifer. > > Juan C. Gutierrez, HT(ASCP) > Histology Laboratory Supervisor > (210)704-2533 > > -----Original Message----- > From: lpwenk@sbcglobal.net [mailto:lpwenk@sbcglobal.net] > Sent: Saturday, September 25, 2004 1:41 PM > To: Jennifer Brielmaier; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Cresyl violet vs. cresyl violet acetate > > I have a question, concerning your "mystery" bottle - > > Do you mean that the teacher has no idea what it is (other than it might be > a cresyl-family dye), and would like some idea/help on how to figure out > what it is? > > Or, is this an assignment, where each student is supposed to find out the > chemical nature of each of the dyes, and thus be able to chemically prove > which dye it is. Where you are getting a grade for this. > > Be honest with us. > > If the teacher actually doesn't know, I think the Histonet community would > be willing to help. > > If this is your homework assignment, I think that the Histonet community > would be willing to refer you to a text book where you could look up the > answers yourself, but that we would not be willing to do your homework for > you. > > Sorry if I'm sounding a little edgy. I'm an instructor in histotechnology. > All of my assignments are designed to teach students about histotechnology. > But there is more to our field than just science. Some of my assignments are > "team" assignments, where my students can pull together their knowledge and > abilities to work on the assignment together. Part of what I'm assess is the > ability of each student to contribute to a team. Some of my other > assignments are "solo", where the person can use book, journals, internet, > etc., but they cannot get the answers from their classmates or other techs. > Part of what I'm assessing is the person's ability to find the answer on > their own, not be told it. I need to know whether a person can problem-solve > and troubleshoot on their own. > > Either way, there are text books and web pages out there with the > information you need about your dyes. I'm sitting here with one of the books > in my lab right now, which has the information. > > So - if your teacher needs the help, I am willing to help and quote from the > pages. > > If you are supposed to find the answer, I'll let you know the name of the > book. And you can look it up for yourself (and hopefully learn about this > book, and learn about other dyes while leafing through this book). > > So, again, be honest, and let me know. > > Peggy A. Wenk, BA, BS, HTL(ASCP)SLS > Program Director > School of Histologic Technicians > School of Histotechnologist > William Beaumont Hospital > Royal Oak, MI 48073 > > ----- Original Message ----- > From: "Jennifer Brielmaier" > To: > Sent: Friday, September 24, 2004 11:43 AM > Subject: [Histonet] Cresyl violet vs. cresyl violet acetate > > > Hello everyone, > > > > I am a first-year graduate student in a biopsychology program and am > enrolled in a basic histology course this semester. Today in class we are > going to learn how to make cresyl violet stain solutions. Our instructor has > informed us that we have a "mystery" bottle in the lab; it is not known > whether it is cresyl violet or cresyl violet acetate. Can anyone tell me > whether there is a simple test that can be performed that will tell us which > solution is in the bottle? Thanks very much. > > > > Jennifer > > > > > > --------------------------------- > > Do you Yahoo!? > > vote.yahoo.com - Register online to vote today! > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Andrew.Prior <@t> Smith-Nephew.com Tue Sep 28 10:58:12 2004 From: Andrew.Prior <@t> Smith-Nephew.com (Prior, Andrew) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Puramatrix gels Message-ID: Has anybody done histology on Puramatrix gels? I am trying to do some histology on cell cultures grown on collagen and Puramatrix gels. The collagen is no problem, but the Puramatrix ix extremely fragile - even changing the culture medium can damage it! I am going to try cryosectioning next week when I get some more gels, but am not hopeful. Any suggestions (not "use another scaffold gel") will be gratefully received. Many thanks 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 gcallis <@t> montana.edu Tue Sep 28 11:07:05 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Cryosectioning versus paraffin sectioning mouse embryo Message-ID: <6.0.0.22.1.20040928095627.01b1a528@gemini.msu.montana.edu> The difference between cryosectioning and paraffin sectioning needs to be clarified. For cryosectioning, you can perfuse a mouse (not sure how this works with pregnant mouse, or do immersion fixation of collected embryos, then cryoprotect with 25 - 30% sucrose overnight, SNAP FREEZE with extreme cold, there have been methods discussed at length on Histonet, then cut sections in a cryostat or cryosectioning. You can also snap freeze fresh, unfixed tissue and do fixation in 4% PFA after the section is cut and mounted on a slide. Paraffin sectioning is taking a fixed embryo, processing it through alcohol gradient, clearing alcohol, and infiltration with paraffin. This sectioning is done on a microtome and IS not cryosectioning. Paraffin would have to be removed from sections prior to staining. You either do one method or the other. but paraffin sections are NOT cut in a cryostat. There are some websites on mouse brain, go to www.mbl.org for mouse brain library. Mouse embryo atlas/photo http://embryo.soad.umich.edu/animal/cdAtlas/cdAtlas.html Embryo, mouse MRI http://www.mouseatlas.caltech.edu/home.html Embryo stage selector http://genex.hgu.mrc.ac.uk/Atlas/intro.html High Resolution Mouse Brain Atlas: http://www.hms.harvard.edu/research/brain/goal.html#top These should at least teach some brain anatomy, and figure our how you want to orient the tissue for final sectioning and the view you need to have. I am going to start processing mouse embryos for in situ and immunohistochemistry, and since I am a novice, I have a few very basic questions that I am hoping to find answers to! First, I was wondering if it is absolutely necessary to embed embryos in paraffin for cryosectioning. I usually work with brain tissue from adult mice, and I never have to embed in paraffin -- I always freeze in OCT. In this case, I will be sectioning embryos at ED14.5 and ED17.5. Also, I was wondering about a simple fixation procedure for 'older' embryos. Is it feasible for fixation to involve only fixing cryosections in 4% paraformaldehyde? Lastly, if my goal is to look at expression patterns in the embryonic brain, what would be the best approach/angle to sectioning? Specifically, I am interested in having a good view of the ventricular zone and dentate gyrus. I would greatly appreciate any and all advice on these matters! Thank you in advance! Ann 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 Sep 28 11:10:56 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] plastic beads and xylene type solvents Message-ID: <6.0.0.22.1.20040928100832.01b1c1a8@gemini.msu.montana.edu> In the world of biomaterials and hard tissue, before any plastic implant is processed through alcohols or any solvent, it is a good practcie to test the solubility of any plastic or other implant in the solvents being used. The same should apply to your plastic fluorophore labelled beads. It would save you a lot of grief and work, not being able to find them if the beads have dissolved away. 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 Sep 28 11:18:29 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] (no subject) Message-ID: <6.0.0.22.1.20040928101101.01b17910@gemini.msu.montana.edu> In reply to using fluorophore labelled primary antibodies, you can still use these for immunohistochemistry if you just come back with a secondary antibody that is anti FITC, antiPE, etc. DAKO has a rabbit antiFITC that is excellent, there are others on the markey. If you want to get away from a biotin systems, you can always use a polymer immunostaining system to detect the rabbit antiFITC. It should work very well. Sorry I can't help you with antibody source. You wrote: Collagen IV subtypes: alpha 3-alpha 6, Does anyone know where I can get antibodies to subtypes of all collagen IV chains (alpha 1?alpha 6)? Are they only available for immunofluorescence detection? SantaCruz has some antibodies for collagen IV alpha 1, alpha 2 and alpha 3 isoforms. I also know that ID Labs in Canada has a clone specific to alpha 1 and alpha 2. 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 dobbin <@t> upei.ca Tue Sep 28 11:42:00 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Histonet and NSH Conventions Message-ID: <41596A27.17470.ED3D8A@localhost> Hello All, I have only been to 2 NSH Conventions (Salt Lake and Toronto) so far, and while I am profoundly impressed with how well they are run and the quality of the speakers, I find that they are sooo big that it is virtually impossible to meet anyone "by chance" that you may know from the Histonet! For instance, the whole time I was in Toronto, I was straining my eyes to read name tags in the hopes of putting faces to some (or any) of the many names I am familiar with here online- and I found none! I would like to suggest that a "Mix and Mingle" opportunity for Histonet members at the annual convention would be a well attended event. An informal gathering, perhaps at lunch time in one of the lecture halls or just around the registration area. And everyone would be asked to wear name tags that have both the 1st and last names nice and big so we wouldn't be a bunch of squinters! :-) Anyone else feel likewise? Comments? Maybe others enjoy the faceless nature of cyberspace!? Cheers! Greg ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 Wanda.Smith <@t> HCAhealthcare.com Tue Sep 28 12:17:20 2004 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Recycled Alocohol and Xylene Message-ID: <8784A3EE34199644AF60DBE517F5B0A604A41864@louex04.lou.medcity.net> Hey Netters, We are demo-ing a recycler and are using the recycled alcohol and xylene on our tissue processor since mid-August. The histotechs here feel that our tissue has recently become dry and brittle and is getting worse. Could this be from the recycled products? We do use brand new reagents in the last 100% and last xylene. Has anyone else experienced this problem using recycled products? Thanks for any insight on this! Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > From JQB7 <@t> CDC.GOV Tue Sep 28 12:19:55 2004 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Histonet and NSH Conventions Message-ID: Someone set up an event like that a few symposiums ago and it was great fun. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Greg Dobbin Sent: Tuesday, September 28, 2004 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histonet and NSH Conventions Hello All, I have only been to 2 NSH Conventions (Salt Lake and Toronto) so far, and while I am profoundly impressed with how well they are run and the quality of the speakers, I find that they are sooo big that it is virtually impossible to meet anyone "by chance" that you may know from the Histonet! For instance, the whole time I was in Toronto, I was straining my eyes to read name tags in the hopes of putting faces to some (or any) of the many names I am familiar with here online- and I found none! I would like to suggest that a "Mix and Mingle" opportunity for Histonet members at the annual convention would be a well attended event. An informal gathering, perhaps at lunch time in one of the lecture halls or just around the registration area. And everyone would be asked to wear name tags that have both the 1st and last names nice and big so we wouldn't be a bunch of squinters! :-) Anyone else feel likewise? Comments? Maybe others enjoy the faceless nature of cyberspace!? Cheers! Greg ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 Wanda.Smith <@t> HCAhealthcare.com Tue Sep 28 12:27:23 2004 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Global Billing Message-ID: <8784A3EE34199644AF60DBE517F5B0A604A41866@louex04.lou.medcity.net> Only got one response...Is anyone out there doing global billing??? Dear Netters, We are in the process of returning to global billing, where the Pathology group bills the patients for the technical and professional fees and pays the hospital for the technical services. Previously, the Pathologist paid the hospital "per block" and per frozen section block, but they did not pay for any special stains, decal charges or multiple slides for levels on biopsy specimens, etc. We would like to be fair, without cutting our own throats! Can anyone that does global billing share the price and billing structure with me? Any help on this would be greatly appreciated! Thanks, Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > From azdudley <@t> hotmail.com Tue Sep 28 14:18:02 2004 From: azdudley <@t> hotmail.com (anita dudley) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Recycled Alocohol and Xylene Message-ID: wanda, we use recycled alcohol on our processor, the xylene is fresh tho. we recycle clearrite on the machine, but use xylene on the processor. haven't had any problems tho with using the recycled alcohol. anita dudley, providence hosp. mobile alabama >From: Smith Wanda >To: "'histonet@lists.utsouthwestern.edu'" > >Subject: [Histonet] Recycled Alocohol and Xylene >Date: Tue, 28 Sep 2004 12:17:20 -0500 > >Hey Netters, >We are demo-ing a recycler and are using the recycled alcohol and xylene on >our tissue processor since mid-August. The histotechs here feel that our >tissue has recently become dry and brittle and is getting worse. Could >this >be from the recycled products? We do use brand new reagents in the last >100% and last xylene. Has anyone else experienced this problem using >recycled products? >Thanks for any insight on this! >Wanda > > > Wanda G. Smith, HTL/HT(ASCP) > > Pathology Supervisor > > Trident Medical Center Laboratory Services > > *9330 Medical Plaza Drive, Charleston, SC 29406 > > *843-797-4586 *fax 843-797-4296 > > *wanda.smith@hcahealthcare.com > > > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee® Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From Jacqueline.Miller <@t> UTSouthwestern.edu Tue Sep 28 14:57:23 2004 From: Jacqueline.Miller <@t> UTSouthwestern.edu (Jacqueline Miller) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] tissue microarray problem Message-ID: Hi everyone, I'm making several practice tissue microarrays, and I'm having a lot of problem with not being able to retrieve the donor core. In other words, after I punch the donor block, the core does not stay in the needle as the needle is withdrawn. Instead, the core remains in the block and the only way to get it out is to pry it out, which ruins the surrounding tissue. I appreciate any tips that anyone can offer as to how to retrieve the core when this happens, or better yet, how to keep this from happening in the first place. Thanks, Jackie From Ben.Shelkowsky <@t> chomp.org Tue Sep 28 15:08:43 2004 From: Ben.Shelkowsky <@t> chomp.org (Shelkowsky, Ben) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Histonet and NSH Conventions Message-ID: <384DA2BD670CE34DA8D3B60F4ED7157F4D6141@exchsrvr.chomp.org> You are 100% on the mark about the difficulty of meeting people at the NSH convention. It might help to make your comments to those that organize the convention (anyone on the convention committee). Years ago when we were meeting in a smaller venue, it was easier. I would suggest a couple of ideas. Join a committee or two. Meet the Regional reps and they'll network with you to meet the state presidents. These folks are our current leaders and you might take the initiative to meet them. They're all there at the hospitalities. Go to the membership meeting. I know it's easier said than done, but it gets easier with each meeting you go to. Good Luck. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bartlett, Jeanine Sent: Tue 9/28/2004 10:19 AM To: Greg Dobbin; histonet@lists.utsouthwestern.edu Cc: Subject: RE: [Histonet] Histonet and NSH Conventions Someone set up an event like that a few symposiums ago and it was great fun. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Greg Dobbin Sent: Tuesday, September 28, 2004 9:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histonet and NSH Conventions Hello All, I have only been to 2 NSH Conventions (Salt Lake and Toronto) so far, and while I am profoundly impressed with how well they are run and the quality of the speakers, I find that they are sooo big that it is virtually impossible to meet anyone "by chance" that you may know from the Histonet! For instance, the whole time I was in Toronto, I was straining my eyes to read name tags in the hopes of putting faces to some (or any) of the many names I am familiar with here online- and I found none! I would like to suggest that a "Mix and Mingle" opportunity for Histonet members at the annual convention would be a well attended event. An informal gathering, perhaps at lunch time in one of the lecture halls or just around the registration area. And everyone would be asked to wear name tags that have both the 1st and last names nice and big so we wouldn't be a bunch of squinters! :-) Anyone else feel likewise? Comments? Maybe others enjoy the faceless nature of cyberspace!? Cheers! Greg ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This is a transmission from Community Hospital of the Monterey Peninsula. This message and any attached documents may be confidential and contain information protected by state and federal medical privacy statutes. They are intended only for the use of the addressee. If you are not the intended recipient, any disclosure, copying, or distribution of this information is strictly prohibited. If you received this transmission in error, please accept our apologies and notify the sender. Thank you. From ajennings <@t> unmc.edu Tue Sep 28 15:10:28 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Histonet and NSH Conventions In-Reply-To: Message-ID: I remember one time, we all met in a big room, that wasn't quite big enough for all of us that showed up. I am not real sure if there was food or drinks even because it was just really fun to put faces to names. I still have my "Histonet" button. I think from two different years. I am willing to be on an informal committee to help organize something for next year in Florida, since I have every intention of attending, but we will need someone who knows the ins and outs of the meeting to head the task. If this doesn?t work, there is also a bulletin board where people post messages if you are looking for someone in particular that you know is attending and they also know about the message board.....and the kiosk for emails that is supplied by a company ( I regrettably don?t remember which one) is the perfect meeting place for histonetters. We have a year...... lets plan From hfedor <@t> jhmi.edu Tue Sep 28 15:27:12 2004 From: hfedor <@t> jhmi.edu (Helen Fedor) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] tissue microarray problem Message-ID: Hi, Move the handle back and forth a couple of times and then gently Tamp on top of the stylus. these two steps have always take care of any resistant core removal. Helen Helen L. Fedor B.S. Johns Hopkins University Pathology Department 600 N Wolfe St Marburg Room 406 Baltimore MD 21287 email: hfedor@jhmi.edu Phone: 410 614-1660 Pager: 410 283-3419 WARNING: E-mail sent over the Internet is not secure. Information sent by e-mail may not remain confidential. DISCLAIMER: This e-mail is intended only for the individual to whom it is addressed. It may be used only in accordance with applicable laws. If you received this e-mail by mistake, notify the sender and destroy the e-mail. >>> Jacqueline Miller 09/28/04 03:57PM >>> Hi everyone, I'm making several practice tissue microarrays, and I'm having a lot of problem with not being able to retrieve the donor core. In other words, after I punch the donor block, the core does not stay in the needle as the needle is withdrawn. Instead, the core remains in the block and the only way to get it out is to pry it out, which ruins the surrounding tissue. I appreciate any tips that anyone can offer as to how to retrieve the core when this happens, or better yet, how to keep this from happening in the first place. Thanks, Jackie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Sep 28 16:01:22 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] NSH conventions and suggestions - who to contact Message-ID: <6.0.0.22.1.20040928145103.01af0588@gemini.msu.montana.edu> Dear all, If you want to make suggestions and comments about the NSH convention/symposium, you should do so to Kerry Crabb, NSH Convention Chairman. He can be contacted at kerry.l.crabb@gsk.com. Kerry and the convention committee are always open to suggestions. I don't think Kerry has time to participate with Histonet, so be sure to email your comments to him (as a CC:) along with your comments to Histonet. He and his committee will appreciate it. As for those who attended the convention, there was a survey you should have filled out with your comments, suggestions, criticisms and turned it. If you still have it, but not filled it out, do so and send to the NSH office, you will find their address at www.nsh.org. This convention survey is read very carefully by the convention committee so they can make future improvements. One new thing this year was the convention notebook was on a CD!! A wonderful way to keep things concise and weightless for those traveling long distances. 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 min_minlu <@t> hotmail.com Tue Sep 28 17:15:40 2004 From: min_minlu <@t> hotmail.com (Min-Min Lu) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] IHC - weak signals Message-ID: TSA ( NEL700A From Perkin Elmer) can be used for amplifying weak signals from IHC Minmin Lu Director, Histology and Gene Expression Core Molecular Cardiology Research Center University of Pennsylvania 982 BRB II 421 Curie Blvd Philadelphia, PA 19104 215-898-0251 >From: "jason m" <kosmicdog@hotmail.com> >To: histonet@pathology.swmed.edu >Subject: [Histonet] IHC - weak signals >Date: Fri, 24 Sep 2004 07:19:25 -0700 > >Does anyone have advice for amplifying weak signals from IHC? Is >there a detection system better than DAB which can make the signal >more noticable? > >_________________________________________________________________ >Designer Mail isn't just fun to send, it's fun to receive. Use >special stationery, fonts and colors. >http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines > Start enjoying all the benefits of MSN® Premium right now and get >the first two months FREE*. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Check out Election 2004 for up-to-date election news, plus voter tools and more! http://special.msn.com/msn/election2004.armx From christopherjackson69 <@t> hotmail.com Tue Sep 28 18:00:46 2004 From: christopherjackson69 <@t> hotmail.com (Christopher Jackson) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Perfusion Fixation of Mouse Hearts Message-ID: <000201c4a5ae$fd84fc20$0302a8c0@CHRISTOPHER> Hi everyone, I am a research assistant at the Ottawa Heart Institute. I am working with mice models. I am supposed to learn how to perfusion fix a mouse heart and then embed it into paraffin wax. Does anyone have a protocol that I could use, or information regarding this technique? Christopher Jackson From jbaez <@t> interscopepath.com Tue Sep 28 18:15:58 2004 From: jbaez <@t> interscopepath.com (Baez, Janet) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] IHC equipment Message-ID: <9E956D8FEB06C2408B08AC16498325E901399B@scopemx1.interscope.com> It's been 5 years since we did IHC in-house. We use to run our IHC on the old TechMate 1000. We're thinking of bringing IHC back in-house. Any feed back as to the new equipment available. Any info will be greatly appreciated. Thanks. Janet E. Baez Histology Manager Interscope Pathology Medical Group 21114 Vanowen St. Canoga Park, Ca. Tel. 818-992-7848 Fax 818-992-6654 From algranth <@t> u.arizona.edu Tue Sep 28 18:43:05 2004 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] NSH conventions and suggestions - who to contact In-Reply-To: <6.0.0.22.1.20040928145103.01af0588@gemini.msu.montana.edu> Message-ID: <4.3.2.7.2.20040928163451.00c72148@algranth.inbox.email.arizona.edu> I'd like to add that when registering for workshops you want to attend at the S/C you consider volunteering to be a liaison or staff assistant. Many of the histonet contributors are also speakers at NSH and this is a great way to meet them and help out the society at the same time. What's in it for you? You could get your workshop free. Andi Grantham At 03:01 PM 9/28/2004 -0600, Gayle Callis wrote: >Dear all, > >If you want to make suggestions and comments about the NSH >convention/symposium, you should do so to Kerry Crabb, NSH Convention >Chairman. He can be contacted at kerry.l.crabb@gsk.com. Kerry and the >convention committee are always open to suggestions. I don't think Kerry >has time to participate with Histonet, so be sure to email your comments >to him (as a CC:) along with your comments to Histonet. He and his >committee will appreciate it. > >As for those who attended the convention, there was a survey you should >have filled out with your comments, suggestions, criticisms and turned >it. If you still have it, but not filled it out, do so and send to the >NSH office, you will find their address at www.nsh.org. This convention >survey is read very carefully by the convention committee so they can make >future improvements. > >One new thing this year was the convention notebook was on a CD!! A >wonderful way to keep things concise and weightless for those traveling >long distances. > >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 ..................................................................... : 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 Jason.PALMER <@t> svhm.org.au Tue Sep 28 20:21:16 2004 From: Jason.PALMER <@t> svhm.org.au (PALMER Jason (SVHM)) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] re vimentin on mouse tissue Message-ID: We use Dako vimentin V9 as a human specific marker in human / mouse hybrid tissue. We have never ever seen it stain mouse tissue of any sort, and that's the way we like it! So, I'm not sure that you'll get it to stain (though I think I remember somebody on histonet in the past claiming that it does work with mouse....). Cheers, Jason Jason Palmer Bernard O'Brien Institute of Microsurgery 42 Fitzroy St, Fitzroy Victoria 3065 Australia tel +61 3 9288 4018 fax +61 3 9416 0926 email: palmerj@svhm.org.au Message: 13 Date: Mon, 27 Sep 2004 15:55:06 -0700 (PDT) From: Yolanda Maldonado Subject: [Histonet] Vimentin on mouse tissue To: histonet@lists.utsouthwestern.edu Message-ID: <20040927225506.66447.qmail@web53507.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi everyone: Does anyone out there has any experience with IHC stain for Vimentin on mouse tissue? I have tried the V9 and 3B4 clone from different companies as well as the ARK kit and other polymers...Nothing seems to work for me!! I have tried EDTA. CB, Reveal, LAB and enzymes as AR and I have also run human and other species along with my tissue to make sure my staining is working. Everything stains but the mice. Any suggestions? -------------- 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 je22r <@t> udcf.gla.ac.uk Wed Sep 29 04:03:07 2004 From: je22r <@t> udcf.gla.ac.uk (Julia Edgar) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] antibody to porcine fibroblasts Message-ID: <004401c4a603$23314240$40e9d182@vet> Dear All Does anybody know of an antibody (preferably a cell surface marker) that will recognise porcine fibroblasts from muscle? Thank you Julia Julia Edgar, BSc (Hons), PhD Institute of Comparative Medicine University of Glasgow Veterinary School From barbara.bublava <@t> meduniwien.ac.at Wed Sep 29 05:09:30 2004 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Thanks Glycerin-Gelatine bubble-phenomen References: <01cd01c49eee$91bc2c50$1401a8c0@GERICHTS9XOZZ8> <1095742272.414fb34026161@webmail.umdnj.edu> Message-ID: <007301c4a60c$69350fc0$1401a8c0@GERICHTS9XOZZ8> I want to thank all the people who responded to my question! What finally did help was the treatment with a mixture of formalin and sulfanilic acid. No more bubbles - even after several days. Thanks a lot Barbara Bublava ----- Original Message ----- From: To: "Barbara Bublava" Cc: Sent: Tuesday, September 21, 2004 1:51 AM Subject: Re: [Histonet] Glycerin-Gelatine bubble-phenomen > Bubbles are caused by desintigration of uncouppled, tissue bound dyes, a > phenomenon described in the 60ties occuring with some enzyme histochemistry > reactions. > A treatment with acidic formalin and sulfanilic acid after the reaction > would reduce/eliminate the bubbling effect. > Regards, > Markus F. Meyenhofer > > > Quoting Barbara Bublava > : > > > Hi Everybody! > > > > I have an interesting phenomen mounting slides with Glyceringelatine > > > > When I mount my slides they do not have bubbles. Oil Red O stains do not > > develop bubbles even after some time but when I mount CAE - Chloracetate > > Esterase (I mount in the same way I do the ORO) the slides develop > > bubbles after some hours and after some days the whole section is covered > > with bubbles, but there are no bubbles around the section! > > > > Why does this happen and how can I prevent this??? > > > > I use Glyceringelatine at 60?C, slides out of dest. water, excess water > > shaked of. I did also try to dry the slides bevore mounting but there > > happend to be more bubbles from the beginning and it did not change this > > strange effect. > > Sometimes, when I am in a hurry I further hardening of the > > Glyceringelatine at fridge (4?C) but I can not see any positive or > > negative effect on bubbles doing that. > > > > Thanks to everyone > > > > Barbara, Vienna > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > -- > From japoteete <@t> saintfrancis.com Wed Sep 29 06:30:57 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] IHC equipment Message-ID: The DAKO autostainers are still work horses and I understand that the new version will be out soon, but I would imagine, rather expensive. I'm wishing! Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Lab Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com > -----Original Message----- > From: Baez, Janet [SMTP:jbaez@interscopepath.com] > Sent: Tuesday, September 28, 2004 6:16 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] IHC equipment > > It's been 5 years since we did IHC in-house. We use to run our IHC on > the old TechMate 1000. We're thinking of bringing IHC back in-house. > Any feed back as to the new equipment available. Any info will be > greatly appreciated. > Thanks. > > Janet E. Baez > Histology Manager > Interscope Pathology Medical Group > 21114 Vanowen St. > Canoga Park, Ca. > Tel. 818-992-7848 > Fax 818-992-6654 > _______________________________________________ > 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 dobbin <@t> upei.ca Wed Sep 29 07:25:11 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Histonet and NSH Conventions In-Reply-To: References: Message-ID: <415A7F76.22305.233265@localhost> Here is how simple it can be: have a table or little kiosk affair set up and call it the Histonet "Out-post" (clever eh?), and that's it! Then Histonetters can stop by the Out-post anytime they wish; to hang out, finish their coffee or whatever and can expect that anyone else they meet there is someone they are probably familiar with from the Histonet! Organizing an "event", even a small one can be a huge task especially at a large convention like ours. I think the "KISS" principle applies here (i.e. keep it simple stupid). And the Out-post idea should not be too burdensome or taxing (with conflicting meetings, etc.), because I think most everyone at least takes coffee breaks! What really sucks about all this, is you all will organize something for next year and by the time I'm able to get to another convention the whole idea will have fizzled again! :-( I think I need a little cheese with that w(h)ine! Have a nice day! Greg To: histonet@lists.utsouthwestern.edu From: ajennings@unmc.edu Date sent: Tue, 28 Sep 2004 15:10:28 -0500 Subject: RE: [Histonet] Histonet and NSH Conventions > > > > > I remember one time, we all met in a big room, that wasn't quite big enough > for all of us that showed up. I am not real sure if there was food or > drinks even because it was just really fun to put faces to names. I still > have my "Histonet" button. I think from two different years. I am willing > to be on an informal committee to help organize something for next year in > Florida, since I have every intention of attending, but we will need > someone who knows the ins and outs of the meeting to head the task. > If this doesn???t work, there is also a bulletin board where people post > messages if you are looking for someone in particular that you know is > attending and they also know about the message board.....and the kiosk for > emails that is supplied by a company ( I regrettably don???t remember which > one) is the perfect meeting place for histonetters. > We have a year...... lets plan ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 kmerriam2003 <@t> yahoo.com Wed Sep 29 07:54:55 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] re vimentin on mouse tissue In-Reply-To: Message-ID: <20040929125455.19201.qmail@web52510.mail.yahoo.com> I agree with Jason. At my former job, we used to use the DAKO V9 clone to stain human cells that had been implanted into mouse tissue. We used it because it did not cross-react with the mouse tissue (we wanted to only see the human cells). Just my 2cents. Kim Merriam Novartis Cambridge, MA "PALMER Jason (SVHM)" wrote: We use Dako vimentin V9 as a human specific marker in human / mouse hybrid tissue. We have never ever seen it stain mouse tissue of any sort, and that's the way we like it! So, I'm not sure that you'll get it to stain (though I think I remember somebody on histonet in the past claiming that it does work with mouse....). Cheers, Jason Jason Palmer Bernard O'Brien Institute of Microsurgery 42 Fitzroy St, Fitzroy Victoria 3065 Australia tel +61 3 9288 4018 fax +61 3 9416 0926 email: palmerj@svhm.org.au Message: 13 Date: Mon, 27 Sep 2004 15:55:06 -0700 (PDT) From: Yolanda Maldonado Subject: [Histonet] Vimentin on mouse tissue To: histonet@lists.utsouthwestern.edu Message-ID: <20040927225506.66447.qmail@web53507.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi everyone: Does anyone out there has any experience with IHC stain for Vimentin on mouse tissue? I have tried the V9 and 3B4 clone from different companies as well as the ARK kit and other polymers...Nothing seems to work for me!! I have tried EDTA. CB, Reveal, LAB and enzymes as AR and I have also run human and other species along with my tissue to make sure my staining is working. Everything stains but the mice. Any suggestions? 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - 100MB free storage! From pmarcum <@t> polysciences.com Wed Sep 29 07:55:27 2004 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Histonet and NSH Conventions In-Reply-To: <415A7F76.22305.233265@localhost> Message-ID: <000e01c4a623$98203410$4f00a8c0@PMARCUM2K> Since several people have made suggestions and most are very good (I like this one) it may be time to ask NSH. We have tables for a number of special groups in the registration area and the biggest problem would be getting volunteers for the table at peak times or having a board where people could not only meet with fellow Histonetters but leave messages to help get together at specific times. This is really up to NSH as an organization. It is really nice to meet the people you talk and see on line in person. Thanks, Pam Marcum? > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Greg > Dobbin > Sent: Wednesday, September 29, 2004 5:25 AM > To: ajennings@unmc.edu > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histonet and NSH Conventions > > > Here is how simple it can be: have a table or little kiosk affair set up > and call it the Histonet "Out-post" (clever eh?), and that's it! Then > Histonetters can stop by the Out-post anytime they wish; to hang > out, finish their coffee or whatever and can expect that anyone else > they meet there is someone they are probably familiar with from the > Histonet! > > Organizing an "event", even a small one can be a huge task > especially at a large convention like ours. I think the "KISS" > principle applies here (i.e. keep it simple stupid). And the Out-post > idea should not be too burdensome or taxing (with conflicting > meetings, etc.), because I think most everyone at least takes coffee > breaks! > > What really sucks about all this, is you all will organize something > for next year and by the time I'm able to get to another convention > the whole idea will have fizzled again! :-( > I think I need a little cheese with that w(h)ine! > Have a nice day! > Greg > > To: histonet@lists.utsouthwestern.edu > From: ajennings@unmc.edu > Date sent: Tue, 28 Sep 2004 15:10:28 -0500 > Subject: RE: [Histonet] Histonet and NSH Conventions > > > > > > > > > > > I remember one time, we all met in a big room, that wasn't > quite big enough > > for all of us that showed up. I am not real sure if there was food or > > drinks even because it was just really fun to put faces to > names. I still > > have my "Histonet" button. I think from two different years. I > am willing > > to be on an informal committee to help organize something for > next year in > > Florida, since I have every intention of attending, but we will need > > someone who knows the ins and outs of the meeting to head the task. > > If this doesn???t work, there is also a bulletin board where > people post > > messages if you are looking for someone in particular that you know is > > attending and they also know about the message board.....and > the kiosk for > > emails that is supplied by a company ( I regrettably don???t > remember which > > one) is the perfect meeting place for histonetters. > > We have a year...... lets plan > > > > > ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ > Greg Dobbin > 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 Luis.Chiriboga <@t> med.nyu.edu Wed Sep 29 07:38:25 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] tissue microarray problem In-Reply-To: Message-ID: couple more ideas...... you want to work with sharp needles, they will get dull over time. If possible make sure that the needle travels all the way through the donor "tissue". This will decrease the likelihood of the tissue core remaining in the block due to it's attachment to surrounding tissue. If your dealing with very fibrous tissue, rotating the entire stylus from left to right when the needle is still in the donor block definitely helps. I have also found that occasionally there is wax build up in the donor needle and that this interferes with removing donor cores. We use the plunger as a ram to keep the bore clean (kind of like cleaning the bore of a cannon or gun with a ram rod in the olden days). Hope this helps Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Helen Fedor Sent: Tuesday, September 28, 2004 4:27 PM To: histonet@lists.utsouthwestern.edu; Jacqueline.Miller@UTSouthwestern.edu Subject: Re: [Histonet] tissue microarray problem Hi, Move the handle back and forth a couple of times and then gently Tamp on top of the stylus. these two steps have always take care of any resistant core removal. Helen Helen L. Fedor B.S. Johns Hopkins University Pathology Department 600 N Wolfe St Marburg Room 406 Baltimore MD 21287 email: hfedor@jhmi.edu Phone: 410 614-1660 Pager: 410 283-3419 WARNING: E-mail sent over the Internet is not secure. Information sent by e-mail may not remain confidential. DISCLAIMER: This e-mail is intended only for the individual to whom it is addressed. It may be used only in accordance with applicable laws. If you received this e-mail by mistake, notify the sender and destroy the e-mail. >>> Jacqueline Miller 09/28/04 03:57PM >>> Hi everyone, I'm making several practice tissue microarrays, and I'm having a lot of problem with not being able to retrieve the donor core. In other words, after I punch the donor block, the core does not stay in the needle as the needle is withdrawn. Instead, the core remains in the block and the only way to get it out is to pry it out, which ruins the surrounding tissue. I appreciate any tips that anyone can offer as to how to retrieve the core when this happens, or better yet, how to keep this from happening in the first place. Thanks, Jackie _______________________________________________ Histonet mailing list Histonet@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 dobbin <@t> upei.ca Wed Sep 29 08:07:46 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Histonet and NSH Conventions In-Reply-To: <000e01c4a623$98203410$4f00a8c0@PMARCUM2K> References: <415A7F76.22305.233265@localhost> Message-ID: <415A8972.25980.4A3282@localhost> Hi Pam, The beauty of the Out-post idea is it would not necessarily require any volunteers. It would just be a meeting place. And you are right, a message board would be useful in the Out-post as well. Greg From: "Pamela Marcum" To: "Greg Dobbin" , Copies to: Subject: RE: [Histonet] Histonet and NSH Conventions Date sent: Wed, 29 Sep 2004 08:55:27 -0400 > Since several people have made suggestions and most are very good (I like > this one) it may be time to ask NSH. We have tables for a number of special > groups in the registration area and the biggest problem would be getting > volunteers for the table at peak times or having a board where people could > not only meet with fellow Histonetters but leave messages to help get > together at specific times. This is really up to NSH as an organization. > It is really nice to meet the people you talk and see on line in person. > > Thanks, > > Pam Marcum? > > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Greg > > Dobbin > > Sent: Wednesday, September 29, 2004 5:25 AM > > To: ajennings@unmc.edu > > Cc: histonet@lists.utsouthwestern.edu > > Subject: RE: [Histonet] Histonet and NSH Conventions > > > > > > Here is how simple it can be: have a table or little kiosk affair set up > > and call it the Histonet "Out-post" (clever eh?), and that's it! Then > > Histonetters can stop by the Out-post anytime they wish; to hang > > out, finish their coffee or whatever and can expect that anyone else > > they meet there is someone they are probably familiar with from the > > Histonet! > > > > Organizing an "event", even a small one can be a huge task > > especially at a large convention like ours. I think the "KISS" > > principle applies here (i.e. keep it simple stupid). And the Out-post > > idea should not be too burdensome or taxing (with conflicting > > meetings, etc.), because I think most everyone at least takes coffee > > breaks! > > > > What really sucks about all this, is you all will organize something > > for next year and by the time I'm able to get to another convention > > the whole idea will have fizzled again! :-( > > I think I need a little cheese with that w(h)ine! > > Have a nice day! > > Greg > > > > To: histonet@lists.utsouthwestern.edu > > From: ajennings@unmc.edu > > Date sent: Tue, 28 Sep 2004 15:10:28 -0500 > > Subject: RE: [Histonet] Histonet and NSH Conventions > > > > > > > > > > > > > > > > > I remember one time, we all met in a big room, that wasn't > > quite big enough > > > for all of us that showed up. I am not real sure if there was food or > > > drinks even because it was just really fun to put faces to > > names. I still > > > have my "Histonet" button. I think from two different years. I > > am willing > > > to be on an informal committee to help organize something for > > next year in > > > Florida, since I have every intention of attending, but we will need > > > someone who knows the ins and outs of the meeting to head the task. > > > If this doesn???t work, there is also a bulletin board where > > people post > > > messages if you are looking for someone in particular that you know is > > > attending and they also know about the message board.....and > > the kiosk for > > > emails that is supplied by a company ( I regrettably don???t > > remember which > > > one) is the perfect meeting place for histonetters. > > > We have a year...... lets plan > > > > > > > > > > ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ > > Greg Dobbin > > 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 > > > ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 JWEEMS <@t> sjha.org Wed Sep 29 08:14:55 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Histonet and NSH Conventions Message-ID: <83AACDB0810528418AA106F9AE9B7F7E278377@sjhaexc02.sjha.org> This is a great idea - sounds fairly simple too. Have a signin book to see who visited. Hopefully I'll get to go next year. Of course I say that every year... and have for the past ten. See you at the Out-post... Happy Wednesday Everyone! j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Greg Dobbin Sent: Wednesday, September 29, 2004 5:25 AM To: ajennings@unmc.edu Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histonet and NSH Conventions Here is how simple it can be: have a table or little kiosk affair set up and call it the Histonet "Out-post" (clever eh?), and that's it! Then Histonetters can stop by the Out-post anytime they wish; to hang out, finish their coffee or whatever and can expect that anyone else they meet there is someone they are probably familiar with from the Histonet! Organizing an "event", even a small one can be a huge task especially at a large convention like ours. I think the "KISS" principle applies here (i.e. keep it simple stupid). And the Out-post idea should not be too burdensome or taxing (with conflicting meetings, etc.), because I think most everyone at least takes coffee breaks! What really sucks about all this, is you all will organize something for next year and by the time I'm able to get to another convention the whole idea will have fizzled again! :-( I think I need a little cheese with that w(h)ine! Have a nice day! Greg To: histonet@lists.utsouthwestern.edu From: ajennings@unmc.edu Date sent: Tue, 28 Sep 2004 15:10:28 -0500 Subject: RE: [Histonet] Histonet and NSH Conventions > > > > > I remember one time, we all met in a big room, that wasn't quite big enough > for all of us that showed up. I am not real sure if there was food or > drinks even because it was just really fun to put faces to names. I still > have my "Histonet" button. I think from two different years. I am willing > to be on an informal committee to help organize something for next year in > Florida, since I have every intention of attending, but we will need > someone who knows the ins and outs of the meeting to head the task. > If this doesn?EUR(tm)t work, there is also a bulletin board where people post > messages if you are looking for someone in particular that you know is > attending and they also know about the message board.....and the kiosk for > emails that is supplied by a company ( I regrettably don?EUR(tm)t remember which > one) is the perfect meeting place for histonetters. > We have a year...... lets plan ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin 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 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 LLElby <@t> LancasterGeneral.org Wed Sep 29 08:37:23 2004 From: LLElby <@t> LancasterGeneral.org (Elby, Lynette L) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Kluver-Barrier Stain Message-ID: Does anyone have the procedure for this stain? If not, can you direct me to where I could find it? Thank in advance..... Lynette Elby Histology Supervisor Lancaster General Hospital 555 North Duke Street Lancaster, PA 17604 (717)544-5065 (717)544-5138 fax (717)305-6144 pager From gcallis <@t> montana.edu Wed Sep 29 09:02:44 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Weak signals Message-ID: <6.0.0.22.1.20040929080110.01b35230@gemini.msu.montana.edu> There are other ways to make weak signals aka enhance the staining. If one gives particulars of what is going on, the exact IHC protocol is needed so suggestions can be made accordingly. TSA or tyramide enhancement is just one of many. 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 Bauer.Karen <@t> mayo.edu Wed Sep 29 09:07:18 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Recycled Alcohol and Xylene Message-ID: <8C6E05FA69571948B461F1327CBB893E0629F17C@lmmail2.ad.lmmhs.org> Hi Wanda, We use recycled xylene in all of our xylene stations on the processor. We also use recycled alcohol in our 70%, 80%, and 95% stations. We have had no problems and our tissues come out nice. Once and a while, really bloody specimens will come out brittle and dry, but a good soak on the ice tray usually fixes that and a nice section can be obtained. Good Luck, Karen Bauer HT(ASCP) Histology Supervisor Department of Pathology Luther Hospital Eau Claire, WI -----Original Message----- From: Smith Wanda [mailto:Wanda.Smith@HCAhealthcare.com] Sent: Tuesday, September 28, 2004 12:17 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Recycled Alocohol and Xylene Hey Netters, We are demo-ing a recycler and are using the recycled alcohol and xylene on our tissue processor since mid-August. The histotechs here feel that our tissue has recently become dry and brittle and is getting worse. Could this be from the recycled products? We do use brand new reagents in the last 100% and last xylene. Has anyone else experienced this problem using recycled products? Thanks for any insight on this! Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From jengirl1014 <@t> yahoo.com Wed Sep 29 09:08:47 2004 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] (no subject) Message-ID: <20040929140847.31686.qmail@web60602.mail.yahoo.com> I'm having a hard time processing kidney, liver and spleen tissue. I do the enter processing the old fashioned way (without a tissue processor). Could I get any suggestions as to what would be a great protocol to use? Thanks in advance! 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!? New and Improved Yahoo! Mail - 100MB free storage! From vazquezr <@t> ohsu.edu Wed Sep 29 09:14:05 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Kluver-Barrier Stain Message-ID: Lynette, I have a Kluver-Barrera method for myelin and nerve cells. Would that be what your looking for? Robyn,OHSU-Portland,Or >>> "Elby, Lynette L" 9/29/2004 6:37:23 AM >>> Does anyone have the procedure for this stain? If not, can you direct me to where I could find it? Thank in advance..... Lynette Elby Histology Supervisor Lancaster General Hospital 555 North Duke Street Lancaster, PA 17604 (717)544-5065 (717)544-5138 fax (717)305-6144 pager _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed Sep 29 09:44:34 2004 From: jkiernan <@t> uwo.ca (J. A. Kiernan) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Malmberg's fixative References: Message-ID: <415ACA52.BE8AE69E@uwo.ca> Is this for preserving vital staining with methylene blue? That's the only use of ammonium picrate that I've come across. Sorry but I cannot find a reference to Malmberg. Someone will! To make a saturated ammonium picrate solution: According to Peter Gray (1954: The Microtomist's Formulary and Guide, p.396-397), add picric acid to a saturated aqueous solution of same, so that plenty of undissolved picric acid is present. Add ammonia, with shaking, drop by drop until the surplus picric acid starts to dissolve. Then add more ammonia until the solution smells of free ammonia. Gray warns of the explosive properties of ammonium picrate; the solution must not be allowed to evaporate to dryness. This is a much more dangerous compound than picric acid (see Merck Index, for example). The compound usually used to immobilize methylene blue in vitally stained tissue is ammonium molybdate, often 6% or so, acidified or in a phosphate-citrate buffer, pH 5.5. This can be followed by a real fixative such as formaldehyde or glutaraldehyde prior to dehydration, clearing etc. A prominent user of vital methylene blue in recent decades has been T. Muller, who has introduced various innovations to the staining method and the fixation that follows, especially for central nervous tissue. Here is a list of some of his papers. Muller T (1989) Paraffin sections of nervous tissue supravitally stained with methylene blue: a new, reliable and simple fixation technique. Stain Technol. 64: 93-96. Muller T (1990) Supravital staining of murine brain with methylene blue according to the Cajal method: a simple and reliable preparation technique. J. Neurosci. Methods 32: 223-226. Muller T (1992) Light-microscopic demonstration of methylene blue accumulation sites in mouse brain after supravital staining. Acta Anat. 144: 39-44. Muller T (1994) Large nerve cells with long axons in the granular layer and white matter of the murine cerebellum. J. Anat. 184: 419-423. Muller T (1994) Supravital uptake of cationic dyes by mast cell granules: a light and electron microscope study. Biotech. Histochem. 69: 171-176. Muller T (1996) The innervation of taste buds in the soft palate of the rat as revealed by methylene blue staining. Arch. Histol. Cytol. 59: 47-54. Muller T (1998) Methylene blue supravital staining: an evaluation of its applicability to the mammalian brain and pineal gland. Histol. Histopathol. 13: 1019-1026. -- ------------------------------- 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 _______________________________ "McCollough, Carol" wrote: > > Greetings Histonetters: > > I have been asked for a recipe for Malmberg's fixative (1:1 ammonium picrate and glycerol). I cannot find a method for the preparation of ammonium picrate. Does anyone have a recipe or a commercial source for this nasty? > > Thanks very much. > > Regards - > Carol > ********************** > Carol B. McCollough, HT/HTL(ASCP) > Diagnostics & Histology Laboratory Manager > Oyster Disease Research Project > Fisheries Service > Maryland Department of Natural Resources > Cooperative Oxford Laboratory > 904 S. Morris Street > Oxford, Maryland 21654 > 410-226-5193 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Wed Sep 29 12:58:38 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Perfusion Fixation of Mouse Hearts In-Reply-To: <000201c4a5ae$fd84fc20$0302a8c0@CHRISTOPHER> References: <000201c4a5ae$fd84fc20$0302a8c0@CHRISTOPHER> Message-ID: <415AF7CE.9030801@umdnj.edu> Hi Christopher: I think the best thing you can do to learn this proceedure is to find a lab doing this and go observe. This is definitely a "picture is worth a thousand words" thing. You could read pages and pages of protocol and still not understand what to do or how to do it. Geoff Christopher Jackson wrote: >Hi everyone, > >I am a research assistant at the Ottawa Heart Institute. I am working >with mice models. > >I am supposed to learn how to perfusion fix a mouse heart and then embed >it into paraffin wax. Does anyone have a protocol that I could use, or >information regarding this technique? > >Christopher Jackson >_______________________________________________ >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 Jacqueline.Miller <@t> UTSouthwestern.edu Wed Sep 29 10:03:40 2004 From: Jacqueline.Miller <@t> UTSouthwestern.edu (Jacqueline Miller) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] tissue microarray problem Message-ID: Thanks for the advice. When you say rotate the entire stylus from left to right, do you mean so that the needle moves as well? I've done that, and once the core is stuck, it actually makes it worse because the needle just moves around the core, making it even harder to get the core to stick. I only have trouble in some areas of some tissue samples (in this case, kidney). I position my donor needle to go as far down as I dare to, and I don't know that our needles would be dull because we've only had this machine for a few months. Again, I appreciate your advice. Any ideas I can get are helpful. Jackie >>> Luis Chiriboga 09/29/04 7:38 AM >>> couple more ideas...... you want to work with sharp needles, they will get dull over time. If possible make sure that the needle travels all the way through the donor "tissue". This will decrease the likelihood of the tissue core remaining in the block due to it's attachment to surrounding tissue. If your dealing with very fibrous tissue, rotating the entire stylus from left to right when the needle is still in the donor block definitely helps. I have also found that occasionally there is wax build up in the donor needle and that this interferes with removing donor cores. We use the plunger as a ram to keep the bore clean (kind of like cleaning the bore of a cannon or gun with a ram rod in the olden days). Hope this helps Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Helen Fedor Sent: Tuesday, September 28, 2004 4:27 PM To: histonet@lists.utsouthwestern.edu; Jacqueline.Miller@UTSouthwestern.edu Subject: Re: [Histonet] tissue microarray problem Hi, Move the handle back and forth a couple of times and then gently Tamp on top of the stylus. these two steps have always take care of any resistant core removal. Helen Helen L. Fedor B.S. Johns Hopkins University Pathology Department 600 N Wolfe St Marburg Room 406 Baltimore MD 21287 email: hfedor@jhmi.edu Phone: 410 614-1660 Pager: 410 283-3419 WARNING: E-mail sent over the Internet is not secure. Information sent by e-mail may not remain confidential. DISCLAIMER: This e-mail is intended only for the individual to whom it is addressed. It may be used only in accordance with applicable laws. If you received this e-mail by mistake, notify the sender and destroy the e-mail. >>> Jacqueline Miller 09/28/04 03:57PM >>> Hi everyone, I'm making several practice tissue microarrays, and I'm having a lot of problem with not being able to retrieve the donor core. In other words, after I punch the donor block, the core does not stay in the needle as the needle is withdrawn. Instead, the core remains in the block and the only way to get it out is to pry it out, which ruins the surrounding tissue. I appreciate any tips that anyone can offer as to how to retrieve the core when this happens, or better yet, how to keep this from happening in the first place. Thanks, Jackie _______________________________________________ Histonet mailing list Histonet@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 Jacqueline.Miller <@t> UTSouthwestern.edu Wed Sep 29 10:06:26 2004 From: Jacqueline.Miller <@t> UTSouthwestern.edu (Jacqueline Miller) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] tissue microarray problem Message-ID: Thanks. I've tried moving the handle back and forth, but that seems to make it worse. I'll have to look into tamping the stylus. I worry about compressing the tissue, but that's what practice is for. Thanks again, Jackie >>> Helen Fedor 09/28/04 3:27 PM >>> Hi, Move the handle back and forth a couple of times and then gently Tamp on top of the stylus. these two steps have always take care of any resistant core removal. Helen Helen L. Fedor B.S. Johns Hopkins University Pathology Department 600 N Wolfe St Marburg Room 406 Baltimore MD 21287 email: hfedor@jhmi.edu Phone: 410 614-1660 Pager: 410 283-3419 WARNING: E-mail sent over the Internet is not secure. Information sent by e-mail may not remain confidential. DISCLAIMER: This e-mail is intended only for the individual to whom it is addressed. It may be used only in accordance with applicable laws. If you received this e-mail by mistake, notify the sender and destroy the e-mail. >>> Jacqueline Miller 09/28/04 03:57PM >>> Hi everyone, I'm making several practice tissue microarrays, and I'm having a lot of problem with not being able to retrieve the donor core. In other words, after I punch the donor block, the core does not stay in the needle as the needle is withdrawn. Instead, the core remains in the block and the only way to get it out is to pry it out, which ruins the surrounding tissue. I appreciate any tips that anyone can offer as to how to retrieve the core when this happens, or better yet, how to keep this from happening in the first place. Thanks, Jackie _______________________________________________ Histonet mailing list Histonet@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 Julie.Sanders <@t> med.va.gov Wed Sep 29 10:01:02 2004 From: Julie.Sanders <@t> med.va.gov (Julie.Sanders@med.va.gov) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] morgue cart covers Message-ID: <457381D92B01BD44B21CF37CC02EBDFD28E95E@vhacinexc2.v10.med.va.gov> I realize this isn't a "real" technical question, but I'm having a very hard time locating a vendor that has plastic covers for morgue carts. The covers we had were about 6X10 ft. and went over a frame that fit on the cart. Does anyone have any idea where to find these? Thanks, Julie Julie Sanders, Supervisor Anatomic Pathology VAMC, Cincinnati, Ohio Julie.Sanders@med.va.gov From ajennings <@t> unmc.edu Wed Sep 29 10:30:42 2004 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] kidney/liver/spleen In-Reply-To: <20040929140847.31686.qmail@web60602.mail.yahoo.com> Message-ID: What problems are you having? Looks like the tissues you are having trouble with are all 'blood' tissues. Human or some other animal? questions size of sample fixed in what for how long? ethyl-alcohols or 'other' for dehydration? do you use xylene or xylene substitute? what volumes are you using to dehydrate and clear? Is the problem noticed before embedding? bottom line..........what is your tissue doing or not doing that constitutes "trouble processing" I'm having a hard time processing kidney, liver and spleen tissue. I do the enter processing the old fashioned way (without a tissue processor). Could I get any suggestions as to what would be a great protocol to use? Thanks in advance! Jennifer K. Sipes, RALAT Sr. Laboratory Technician Johns Hopkins University From algranth <@t> u.arizona.edu Wed Sep 29 10:38:04 2004 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Bouins substitute in Trichrome staining Message-ID: <4.3.2.7.2.20040929083201.00c72f08@algranth.inbox.email.arizona.edu> I attended a workshop at NSH last week on connective tissue staining and one of the attendees said they were using citrate buffer instead of bouins as a mordant. I couldn't catch up with her after the workshop to ask about the protocol. Is anybody out in histoland doing this? Would you mind sharing your protocol if you are? 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 jannanunez <@t> hotmail.com Wed Sep 29 10:30:47 2004 From: jannanunez <@t> hotmail.com (Janna Nunez-Gussman) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Please unsubscribe me from your mailing list. Thanks Message-ID: Please unsubscribe me from your mailing list. Thanks! Janna _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From JWEEMS <@t> sjha.org Wed Sep 29 10:46:42 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] morgue cart covers Message-ID: <83AACDB0810528418AA106F9AE9B7F7E27837C@sjhaexc02.sjha.org> THermo Shandon - Cat # 8253.. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Julie.Sanders@med.va.gov Sent: Wednesday, September 29, 2004 11:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] morgue cart covers I realize this isn't a "real" technical question, but I'm having a very hard time locating a vendor that has plastic covers for morgue carts. The covers we had were about 6X10 ft. and went over a frame that fit on the cart. Does anyone have any idea where to find these? Thanks, Julie Julie Sanders, Supervisor Anatomic Pathology VAMC, Cincinnati, Ohio Julie.Sanders@med.va.gov _______________________________________________ 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 Wanda.Smith <@t> HCAhealthcare.com Wed Sep 29 10:45:54 2004 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] morgue cart covers Message-ID: <8784A3EE34199644AF60DBE517F5B0A604A4186D@louex04.lou.medcity.net> Julie, Where I worked before, the gentleman in charge of the morgue had an auto upholstery place make heavy vinyl covers for the carts used to transport the bodies from the floor. If you take the old cover, I'm sure they could make a pattern by it. They seem to last a lot longer than the cover that came with the cart. Hope that helps! Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > -----Original Message----- From: Julie.Sanders@med.va.gov [mailto:Julie.Sanders@med.va.gov] Sent: Wednesday, September 29, 2004 11:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] morgue cart covers I realize this isn't a "real" technical question, but I'm having a very hard time locating a vendor that has plastic covers for morgue carts. The covers we had were about 6X10 ft. and went over a frame that fit on the cart. Does anyone have any idea where to find these? Thanks, Julie Julie Sanders, Supervisor Anatomic Pathology VAMC, Cincinnati, Ohio Julie.Sanders@med.va.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From shive003 <@t> umn.edu Wed Sep 29 10:49:10 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] slighty off-topic question for IHC workers Message-ID: <006e01c4a63b$dc348fd0$78065486@vdl220FAC> My personal doctor has asked me to pose this question to others in my professional field. I have some interesting medical history I'd like to share with you, and am asking for any comments you may have (and I apologize ahead of time if you think this is an inappropriate forum in which to broach this subject). Four years ago, after a routine health check-up, my bloodwork came back with a positive FANA test. My general practitioner referred me to a rheumatologist for follow-up, saying that I possibly had lupus erythematosis. The subsequent follow-up proved negative, since I had no other signs nor symptoms of the disease, and no other abnormal bloodwork. Then, one month ago, again a routine health check-up came up with abnormal bloodwork results. But this time, the FANA and other CBC values were normal; however, the rheumatoid factor was now TEN times the normal level, indicating I might have rheumatoid arthritis. Again, after a trip to the rheumatologist, the answer was the same... I did not have any other signs or symptoms of RA. So, the rheumatologist is stymied... and his curiosity was piqued when I told him I've been working with antibodies and animal sera for 20 years (and many of those times without gloves, tsk tsk). He's wondering if there might be some passive absorption of antibodies/animal proteins into my system and my body is mounting a wacky response to them. At least this is what I gathered from his query. Has anyone had similar experiences that I can relay to him as anecdotal information? And before you all send those thousands of get-well cards, I'm in very fine health otherwise! Thanks for any input you may have. Jan Shivers IHC Lab U of MN Vet Diag Lab St. Paul, MN, USA From lizchlipala <@t> premierhistology.com Wed Sep 29 10:51:04 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] morgue cart covers In-Reply-To: <457381D92B01BD44B21CF37CC02EBDFD28E95E@vhacinexc2.v10.med.va.gov> Message-ID: <000601c4a63c$20b30bf0$76d48a80@AMY> MOPEC sells them. I have bought them from them is the past and they are very nice. They can make them to fit any size. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology 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 Julie.Sanders@med.va.gov Sent: Wednesday, September 29, 2004 8:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] morgue cart covers I realize this isn't a "real" technical question, but I'm having a very hard time locating a vendor that has plastic covers for morgue carts. The covers we had were about 6X10 ft. and went over a frame that fit on the cart. Does anyone have any idea where to find these? Thanks, Julie Julie Sanders, Supervisor Anatomic Pathology VAMC, Cincinnati, Ohio Julie.Sanders@med.va.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tissuearray <@t> hotmail.com Wed Sep 29 10:52:32 2004 From: tissuearray <@t> hotmail.com (Thom Jensen) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Fixing tissue microarray problem Message-ID: If the tissue is deep you might not be going entirely through it to the paraffin on the other side. Even so sometimes the core still stays in. The habit I have when punching is turning the donor block around and then removing the needle. This complete turn seems to be the best remedy for stuck cores. Thom Jensen for more info on Tissue Microarray Instruction visit: www.arrayworkshop.com >From: "Jacqueline Miller" >To: , >Subject: Re: [Histonet] tissue microarray problem >Date: Wed, 29 Sep 2004 10:06:26 -0500 > >Thanks. I've tried moving the handle back and forth, but that seems to >make it worse. I'll have to look into tamping the stylus. I worry >about compressing the tissue, but that's what practice is for. > >Thanks again, >Jackie > >>> Helen Fedor 09/28/04 3:27 PM >>> >Hi, >Move the handle back and forth a couple of times and then gently Tamp >on top of the stylus. these two steps have always take care of any >resistant core removal. >Helen > >Helen L. Fedor B.S. >Johns Hopkins University >Pathology Department >600 N Wolfe St >Marburg Room 406 >Baltimore MD 21287 >email: hfedor@jhmi.edu >Phone: 410 614-1660 >Pager: 410 283-3419 > >WARNING: E-mail sent over the Internet is not secure. Information sent >by e-mail may not remain confidential. >DISCLAIMER: This e-mail is intended only for the individual to whom it >is addressed. It may be used only in accordance with applicable laws. If >you received this e-mail by mistake, notify the sender and destroy the >e-mail. > > > > >>> Jacqueline Miller 09/28/04 >03:57PM >>> >Hi everyone, > >I'm making several practice tissue microarrays, and I'm having a lot >of >problem with not being able to retrieve the donor core. In other >words, >after I punch the donor block, the core does not stay in the needle as >the needle is withdrawn. Instead, the core remains in the block and >the >only way to get it out is to pry it out, which ruins the surrounding >tissue. I appreciate any tips that anyone can offer as to how to >retrieve the core when this happens, or better yet, how to keep this >from happening in the first place. > >Thanks, >Jackie > >_______________________________________________ >Histonet mailing list >Histonet@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 _________________________________________________________________ [1]Get ready for school! Find articles, homework help and more in the Back to School Guide! References 1. http://g.msn.com/8HMBENUS/2743??PS=47575 From GauchV <@t> mail.amc.edu Wed Sep 29 10:54:37 2004 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Barbitol Message-ID: Does anyone know what we can substitute for Barbitol in ATPase buffers for muscle stains? Any help you can give me would be greatly appreciated.. Thanks, Vicki Gauch AMCH Albany, NY From jfish <@t> gladstone.ucsf.edu Wed Sep 29 11:23:11 2004 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Histonet and NSH Conventions In-Reply-To: <415A7F76.22305.233265@localhost> References: <415A7F76.22305.233265@localhost> Message-ID: Why not ask NSH to add an extra line to our name tags, it could be either your email address, or the name that shows up in the "who" column in our email box. That way we can place the name (or email name) with the face. The last convention I attended I spent most of my time staring at name tags to see who, where, and what about each person. Mostly to see if I recognized anyone from the list. It is somewhat of a thrill to recognize a name as I have probably gleened a small bit of information from them via the histonet. But the problem is that many of you don't show your name on the email, only your address. Hint, hint! Just a thought. Jo Dee ********************************************************************** ********** Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease Telephone: (415) 695-3720 Fax: (415) 285-5632 E-mail: jfish@gladstone.ucsf.edu Mailing address: Gladstone Institutes P.O. Box 419100 San Francisco, CA 94141-9100 From Cathy.Stevens <@t> HealthONEcares.com Wed Sep 29 11:52:04 2004 From: Cathy.Stevens <@t> HealthONEcares.com (Stevens Cathy) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Recycled Alocohol and Xylene Message-ID: We have been recycling for 3 years without any problems. We use xylene (recycled) and recycled 70%, 95% on our processor. The only thing not recycled is the absolute alcohol. We have a B/R and really like it. -----Original Message----- From: Smith Wanda [mailto:Wanda.Smith@HCAhealthcare.com] Sent: Tuesday, September 28, 2004 11:17 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Recycled Alocohol and Xylene Hey Netters, We are demo-ing a recycler and are using the recycled alcohol and xylene on our tissue processor since mid-August. The histotechs here feel that our tissue has recently become dry and brittle and is getting worse. Could this be from the recycled products? We do use brand new reagents in the last 100% and last xylene. Has anyone else experienced this problem using recycled products? Thanks for any insight on this! Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From northma <@t> ohsu.edu Wed Sep 29 12:01:18 2004 From: northma <@t> ohsu.edu (Mary North) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Kluver-Barrier Stain Message-ID: The method for Luxol Fast Blue (Kluver-Barrera) Method for myelin and nerve cells can be found in the AFIP Laboratory Methods in Histotechnology staining manuals. Mary North, HT(ASCP)HTL Oregon Health & Science University Portland, OR >>> "Elby, Lynette L" 9/29/2004 6:37:23 AM >>> Does anyone have the procedure for this stain? If not, can you direct me to where I could find it? Thank in advance..... Lynette Elby Histology Supervisor Lancaster General Hospital 555 North Duke Street Lancaster, PA 17604 (717)544-5065 (717)544-5138 fax (717)305-6144 pager _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From psanquin <@t> lugo.usc.es Wed Sep 29 12:14:00 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Sucrose cryoprection Message-ID: <3.0.6.32.20040929191400.007f7ae0@pop.lugo.usc.es> Dear listers, I am cutting at the sliding microtome fetal and early postantal mice brains. Could somebody tell me if is it normal that after sucrose cryoprection the pieces of tissue show a gelatin-like appearance? I have lot of problems cutting this tissue. Thanks in advance Pablo From jengirl1014 <@t> yahoo.com Wed Sep 29 12:20:23 2004 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] (no subject) In-Reply-To: <001801c4a62e$545b1be0$4f00a8c0@PMARCUM2K> Message-ID: <20040929172023.78314.qmail@web60602.mail.yahoo.com> It's mouse tissue. Here's the way I've been doing it recently. I've made changes from my original protocol. Flex 70 (made from Flex 80).........1 hour (2 times) Flex 95.......................................45 min Flex 95.......................................1 hr Flex 100.....................................1 hr (2 times) ClearRite 3..................................1 hr @ Room Temp ClearRite 3..................................1 hr @ 60 degrees ClearRite 3/Parafin.......................1 hr @ 60 degrees Parafin........................................Overnight (It's my stopping point) Parafin........................................1 hr Parafin........................................1 hr Please let me know if I need to change anything. 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!? vote.yahoo.com - Register online to vote today! From Janet.Bonner <@t> FLHOSP.ORG Wed Sep 29 12:35:20 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Barbital Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4080@fh2k093.fhmis.net> We use 0.1M Glycine Buffer with 0.75M Calcium Chloride: 0.1M Glycine Buffer: Glycine...........750 mg Sodium chloride...585 mg Distilled water...100 ml Stable for 6 months at 4c 0.75M Calcium Chloride, Dihydrite: Calcium Chloride, dihydrite....11.03 gm Distilled water...............100 ml 0.1M Sodium Hydroxide: Sodium Hydroxide.........0.4gm ____________ Distilled water..........100 ml 0.1M Glycine Buffer with 0.75M Calcium Chloride 0.1M Glycine buffer..........50 ml 0.75M Calcium Chloride........10 ml 0.1M Sodium hydroxide........22 ml Adjust pH to 9.4 with additional sodium hydroxide as necessary. As the pH increases, the calcium precipitates. Let the precipitate settle to the bottom or filter before use. May be stored for 6 months at 4C. Warm to room temperature and check pH before use. Incubation Solution: Adenosine triphosphate(ATP)................5 mg 0.1M glycine buffer with calcium chloride..10 mg Mix and adjust pH to 9.4 with 0.1M sodium hydroxide or 0.1M hydrochloric acid. Our main reference is Frieda Carson's HISTOTECHNOLOGY:A SELF INSTRUCTIONAL TEXT, second edition, pp 260-262, pp264-265. -----Original Message----- From: Vicki Gauch [mailto:GauchV@mail.amc.edu] Sent: Wednesday, September 29, 2004 11:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Barbitol Does anyone know what we can substitute for Barbitol in ATPase buffers for muscle stains? Any help you can give me would be greatly appreciated.. Thanks, Vicki Gauch AMCH Albany, NY _______________________________________________ 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 vazquezr <@t> ohsu.edu Wed Sep 29 13:12:40 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Kluver-Barrier Stain Message-ID: Lynette, I have the manual, if you would like, I can fax the procedure to you? Robyn OHSU,Portland,Or >>> "Mary North" 9/29/2004 10:01:18 AM >>> The method for Luxol Fast Blue (Kluver-Barrera) Method for myelin and nerve cells can be found in the AFIP Laboratory Methods in Histotechnology staining manuals. Mary North, HT(ASCP)HTL Oregon Health & Science University Portland, OR >>> "Elby, Lynette L" 9/29/2004 6:37:23 AM >>> Does anyone have the procedure for this stain? If not, can you direct me to where I could find it? Thank in advance..... Lynette Elby Histology Supervisor Lancaster General Hospital 555 North Duke Street Lancaster, PA 17604 (717)544-5065 (717)544-5138 fax (717)305-6144 pager _______________________________________________ Histonet mailing list Histonet@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 northma <@t> ohsu.edu Wed Sep 29 13:19:10 2004 From: northma <@t> ohsu.edu (Mary North) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Barbitol Message-ID: We have success using Sigma's Alkaline buffer solution, 1.5 M. Catalog # A9226. If you want, I can attach the procedure to your email address. Mary North, HT(ASCP)HTL Neuromuscular Lab OHSU Portland, OR >>> "Vicki Gauch" 9/29/2004 8:54:37 AM >>> Does anyone know what we can substitute for Barbitol in ATPase buffers for muscle stains? Any help you can give me would be greatly appreciated.. Thanks, Vicki Gauch AMCH Albany, NY _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Sep 29 13:21:14 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] Hand processing mouse tissues In-Reply-To: <20040929172023.78314.qmail@web60602.mail.yahoo.com> References: <001801c4a62e$545b1be0$4f00a8c0@PMARCUM2K> <20040929172023.78314.qmail@web60602.mail.yahoo.com> Message-ID: <6.0.0.22.1.20040929120835.01b26728@gemini.msu.montana.edu> First of all you should invest in three things, a vacuum oven that heats, and a Wheaton O ring vacuum dessictor and a vacuum pump. You can do mouse tissues in one day with the following schedule.. My thoughts are you are overdehydrating and exposing your mouse tissues to too much heat. Fixation is complete. Use the vacuum dessicator, Wheaton O ring is perfect, no grease is needed. Do all steps under vacuum at 20 psi. Flex 70 30 min Flex 95 30 min Flex 95 30 min Flex 100 30 min Flex 100 30 min Clearite 30 to 45 min RT Clearite 3 30 to 45 min RT You do not need a Clearite/paraffin mixture! And do not add heat to Clearite steps!! You are probably cooking your tissues with too much heat, particularly overnight in paraffin. Paraffin 30 min no more than 60C. Paraffin 30 min Paraffin 30 min Paraffin 30 min (no more than two hours total in paraffin) This is a 5 1/2 hour schedule, within a working day. The inexpensive items for having vacuum in all steps will speed up your processing of lean, tiny mouse tissues. You did not say, but I am reading between the lines your tissues are very dry and friable. If your tissues seem a bit under-dehydrated, increase time in one 95% and in one 100% to 45 min. Be aware of what tissues you are processing, you may want to change the schedule a bit for denser, or larger pieces, liver, whole brains, etc. Tiny nodes will take possibly even less time. Use a vacuum oven for all your paraffin infiltrations. Just have beakers of melted paraffin, drop cassettes into this, hopefully you have a wire/mesh basket to speed up transfer of cassettes, an old Technicon basket will work nicely. or tie them into some cheesecloth for carrying from one container to another. At 11:20 AM 9/29/2004, you wrote: >It's mouse tissue. Here's the way I've been doing it recently. I've made >changes from my original protocol. > >Flex 70 (made from Flex 80).........1 hour (2 times) >Flex 95.......................................45 min >Flex 95.......................................1 hr >Flex 100.....................................1 hr (2 times) >ClearRite 3..................................1 hr @ Room Temp >ClearRite 3..................................1 hr @ 60 degrees >ClearRite 3/Parafin.......................1 hr @ 60 degrees >Parafin........................................Overnight (It's my stopping >point) >Parafin........................................1 hr >Parafin........................................1 hr > >Please let me know if I need to change anything. > > >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!? >vote.yahoo.com - Register online to vote 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 sa.drew <@t> hosp.wisc.edu Wed Sep 29 13:48:02 2004 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] (no subject) Message-ID: Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 From sa.drew <@t> hosp.wisc.edu Wed Sep 29 13:50:21 2004 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:24:05 2005 Subject: [Histonet] (no subject) Message-ID: Has anyone tried the Anti-SV40 T Antigen(AB-3) from Calbiochem/Onogene? We need to replace our polyoma virus antibody... Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 From hymclab <@t> hyhc.com Wed Sep 29 14:23:19 2004 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Bouins substitute in Trichrome staining Message-ID: PLEASE POST -----Original Message----- From: Andrea Grantham [mailto:algranth@u.arizona.edu] Sent: Wednesday, September 29, 2004 10:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bouins substitute in Trichrome staining I attended a workshop at NSH last week on connective tissue staining and one of the attendees said they were using citrate buffer instead of bouins as a mordant. I couldn't catch up with her after the workshop to ask about the protocol. Is anybody out in histoland doing this? Would you mind sharing your protocol if you are? 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 histophilhuff <@t> yahoo.com Wed Sep 29 15:17:44 2004 From: histophilhuff <@t> yahoo.com (Phillip Huff) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Perfusion Fixation of Mouse Hearts In-Reply-To: <000201c4a5ae$fd84fc20$0302a8c0@CHRISTOPHER> Message-ID: <20040929201744.61127.qmail@web50305.mail.yahoo.com> We currently perform routine mouse perfusions using paraformaldehyde as our fixative. We place a needle into the left ventricle of the heart and perfuse at 120 mmHg for 2-3 min, however we need to clamp the heart to the needle. I can forward you our detailed protocol for mouse perfusions, however you may have to adapt the procedure depending on the area of the heart you are interesting in analyzing. If a small hole at the apex of the heart is no problem then this protocol should work for you. Phil Christopher Jackson wrote: Hi everyone, I am a research assistant at the Ottawa Heart Institute. I am working with mice models. I am supposed to learn how to perfusion fix a mouse heart and then embed it into paraffin wax. Does anyone have a protocol that I could use, or information regarding this technique? Christopher Jackson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Mail - 50x more storage than other providers! From jgjulander <@t> cc.usu.edu Wed Sep 29 16:06:08 2004 From: jgjulander <@t> cc.usu.edu (Justin Julander) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Help me get rid of autofluorescence Message-ID: <415B2A46@webster.usu.edu> I have a virus that expresses GFP during replication. I have fixed tissue from animals infected with the virus in 4% paraformaldehyde (Paraformaldehyde from Ted Pella as a 16% solution) in water. I fix tissues for 4 hours, soak 1-2 days in 30% sucrose, freeze in OCT, and then section on the cryostat. My control uninfected tissue samples have high fluorescence. Is there a way to avoid this autofluorescence? Keep in mind that I have to fix the samples to kill the virus (dangerous to man). Thanks in advance, Justin Institute for Antiviral Research Biotechnology Center Rm. 201 4700 Old Main Hill Utah State University Logan, UT 84322-4700 Fax (435)797-3959 Ph. (435)797-3643 jgjulander@cc.usu.edu From Jackie.O'Connor <@t> abbott.com Wed Sep 29 16:25:56 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] T-Box2 antibody search Message-ID: Hi y'all - (sorry, my daughter's boyfriend from Texas was here visiting) Anyway - I'm looking for T-Box2 for IHC - any tips?? Jackie O' From gcallis <@t> montana.edu Wed Sep 29 16:41:59 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Help me get rid of autofluorescence In-Reply-To: <415B2A46@webster.usu.edu> References: <415B2A46@webster.usu.edu> Message-ID: <6.0.0.22.1.20040929153653.01b34d30@gemini.msu.montana.edu> Unfortunately, you have to live with it. Anytime you fix tissue with paraformaldehyde or NBF, you will increase autofluorescence. Sometimes it is better to snap freeze fresh tissue,cryosection and fix with formalin vapors. Obviously, this is not possible for you, OR you can detect the GFP with immunohistochemistry or immunofluorescence using a different colored fluorophore. This was discussed this past summer at great length with one gentleman from Germany who succeeded in using the formalin vapor method. I think I also have that as a pdf and will send to you. Not sure about DsRed, but maybe a red GFP chimera is a better choice and use the autofluorescence like a counterstain??? I have a review of autofluorescence in conjunction with GFP I am going to attach to you privately. A very interesting paper. At 03:06 PM 9/29/2004, you wrote: >I have a virus that expresses GFP during replication. I have fixed tissue >from animals >infected with the virus in 4% paraformaldehyde (Paraformaldehyde from Ted >Pella as a >16% solution) in water. I fix tissues for 4 hours, soak 1-2 days in 30% >sucrose, freeze in >OCT, and then section on the cryostat. My control uninfected tissue samples >have high >fluorescence. Is there a way to avoid this autofluorescence? Keep in mind >that I have to >fix the samples to kill the virus (dangerous to man). >Thanks in advance, >Justin > >Institute for Antiviral Research >Biotechnology Center Rm. 201 >4700 Old Main Hill >Utah State University >Logan, UT 84322-4700 >Fax (435)797-3959 >Ph. (435)797-3643 >jgjulander@cc.usu.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 Laurie.Pereira <@t> sdcounty.ca.gov Wed Sep 29 16:53:12 2004 From: Laurie.Pereira <@t> sdcounty.ca.gov (Pereira, Laurie ) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] re: salaries Message-ID: Thanks to all who have enlightened me on the pros and cons of moving to Florida :) Yes, I'm still leaving but at least I know a little bit about the salary differences from San Diego to Florida. I still need to do a little more research on the differences of cost of living but I've got some time. Thanks again!! Laurie From vazquezr <@t> ohsu.edu Wed Sep 29 17:26:52 2004 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] re: salaries Message-ID: Laurie, So, what is the pay rate there in Florida? If you don't mind me asking... Robyn OHSU,Derm Surg,Portland,Or >>> "Pereira, Laurie " 9/29/2004 2:53:12 PM >>> Thanks to all who have enlightened me on the pros and cons of moving to Florida :) Yes, I'm still leaving but at least I know a little bit about the salary differences from San Diego to Florida. I still need to do a little more research on the differences of cost of living but I've got some time. Thanks again!! Laurie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From aldo.anile <@t> HDScientific.com.au Wed Sep 29 18:08:57 2004 From: aldo.anile <@t> HDScientific.com.au (Aldo Anile) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] RE: RE: Recycled Alcohol and Xylene Message-ID: Just a passing comment. When recycling xylene, in most cases the purity is in fact greater than the xylene you commercially buy, so using recycled xylene should not impact at all on processing or staining. When recycling alcohol, because of the chemistry involved in the distillation and the affinities of alcohol, at best only approx 96% alcohol is ever achieved through recyling. If using recycled alcohol in processing I suggest several changes of absolute alcohol at the end of your dehydration step. Recycled alcohol is best only used up to the 95% alcohol dehydrating steps in processing. Aldo Anile Applications Consultant HD SCIENTIFIC SUPPLIES PTY LTD Phone: + 61 3 9364 9569 Mobile: 0408 471 485 E-mail: aldo.anile@hdscientific.com.au Web: www.hdscientific.com.au From Laurie.Pereira <@t> sdcounty.ca.gov Wed Sep 29 18:54:01 2004 From: Laurie.Pereira <@t> sdcounty.ca.gov (Pereira, Laurie ) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] re:salaries Message-ID: The pay rate varies for histotechnicians. Anywhere between $12-24 per hour depending on experience and location. From RFail <@t> Charleston.net Wed Sep 29 20:30:04 2004 From: RFail <@t> Charleston.net (Rena) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Red chromogens Message-ID: <000001c4a68d$03c24780$c211a6a5@renad4yk9b8abe> I have been having some trouble with some Abs stained with New Fuchsin and permanent red. Recently I was asked if I were using Levamisole in the permanent red and if so how much? I was told too much Levamisole in permanent red would result in no staining. I have used 1 drop per ml for both New Fuchsin and permanent red, which is the appropriate amount. We have run both DABS and either permanent red or New Fuchsin with the same AB from the same vial with vastly different results. Any ideas? Rena Fail From jkiernan <@t> uwo.ca Wed Sep 29 22:29:07 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Red chromogens [alkaline phosphatase labels] References: <000001c4a68d$03c24780$c211a6a5@renad4yk9b8abe> Message-ID: <415B7D83.64C14C0A@uwo.ca> Dear Rena Fail, Red alkaline phosphatase product. Methods giving permanently mountable red products have been available for many years. Davis & Ornstein (1959) introduced hexazotized pararosaniline as a trifunctional trapping agent for naphthols released by hydrolysis of naphthol phosphates. Pararosaniline (CI 42500; CI Basic red 9) is one of the four components of basic fuchsine and is commercially available in fairly pure form. New fuchsine (CI 42520; CI Basic violet 2), which is closely similar to pararosaniline and also available in pure form (Horobin, 2002); It is currently preferred for making the hexazotized reagent. Kits are sold, but the substrate solution is easily made in the lab. In a simple procedure published at the IHCWorld.com (2004) web site (see below for its URL), the working mixture is prepared from four stock solutions, which are stored at 4 C. Solution D is warmed to room temperature before using. The amount of levamisole (inhibitor of endogenous tissue alkaline phosphatase) is clearly stated. The working solution is made immediately before using. Mix 10 drops each of A and B, then add 10 drops of solution C followed by 10 Kits are sold, but the substrate solution is easily made in the lab. In a simple procedure published at the IHCWorld.com (2004) web site, the working mixture is prepared from four stock solutions, which are stored at 4 C. Solution D is warmed to room temperature before using. I've annotated the instructions from IHCworld.com to simplify the weighing and measuring. A. New fuchsine (CI 42520), 0.2% in 2M HCl. (Conc. HCl is 10 or 12M) B. Sodium nitrite (NaNO2), 0.4% and levamisole,(= [?]-tetramisole hydrochloride) 0.48% in water. C. Naphthol AS-BI phosphate, 1% in 100% dimethylformamide. D. 0.05 M Tris-HCl buffer, pH 8.7. The working solution is made immediately before using. Mix 10 drops each of A and B, then add 10 drops of solution C followed by 10ml of the buffer (D). A drop is assumed to be 0.05 ml. Sections are incubated in the working solution for 10-20 minutes at room temperature. They are then washed counterstained as desired, dehydrated, cleared and coverslipped. Lojda et al (1979, p. 74-75) warn that the working solution may be red or brown from colored impurities in new fuchsine or pararosaniline. Such solutions cause excessive yellow background staining. The yellow/brown impurities are the same ones that can make Schiff's reagent yellow (removable from Schiff with activated charcoal). Pararosaniline certified by the Biological Stain Commission is suitable for making alkaline phosphatase substrate mixtures because it is required to be substantially free of brown and yellow materials (Penney et al., 2002). Probably any batch of basic fuchsine certified by the Biological Stain Commission will be OK for making the IHCWorld.com alkaline phosphatase substrate solution, because each of the four possible components of basic fuchsine has three diazotizable amino groups and their molecular weights are all quite close. I don't know of any published study that establishes this, so you're probably safest to go with either certified pararosaniline or with new fuchsine from a reputable dealer. References. (Please check them out if you can. The 2nd is easy enough!) Davis & Ornstein (1959) J. Histochem. Cytochem. 7:297-298. IHCWorld.com (2004) New fuchsin alkaline phosphatase substrate solution. http://www.ihcworld.com/_protocols/chromogen_substrates/AP_new_fuchsin_red.htm Horobin (2002) Chapter 14 in Conn's Biological Stains, 10th ed. Oxford:BIOS. Lojda et al (1979) Enzyme Histochemistry. Berlin: Springer. Penney et al. (2002) Biotech. Histochem. 77:237-275. Finally, please note that the correct spelling is fuchsine, not fuchsin, despite anything you might read in a catalog or on a bottle label. This is not a matter of American, British and German spellings. (Fuchsin is German for vixen; the dyes are, I think, named from their colours being similar to those of Fuchsia flowers rather than foxes.) Look in Websters, the Oxford, etc. It's fuchsine with an e. An -ine ending of an informal name of a chemical indicates that it is a base, often an amine. The -in ending occurs with compounds that are not bases, such as dextrin, eosin etc. John Kiernan London, Canada ------------------------------------------------------------------ Rena wrote: > > I have been having some trouble with some Abs stained with New Fuchsin > and permanent red. Recently I was asked if I were using Levamisole in > the permanent red and if so how much? I was told too much Levamisole in > permanent red would result in no staining. I have used 1 drop per ml for > both New Fuchsin and permanent red, which is the appropriate amount. We > have run both DABS and either permanent red or New Fuchsin with the > same AB from the same vial with vastly different results. Any ideas? > Rena Fail > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RFail <@t> Charleston.net Thu Sep 30 03:50:14 2004 From: RFail <@t> Charleston.net (Rena) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] RE:Vixens/ Red chromogens [alkaline phosphatase labels] In-Reply-To: <415B7D83.64C14C0A@uwo.ca> Message-ID: <000001c4a6ca$830e65e0$8711a6a5@renad4yk9b8abe> Dear John Kiernan, I really appreciate your help regarding a permanently mountable red product, as always your response was thorough and informative. I was particularly interested in the last bit of information and shall correct my spelling of fuchsine so that the dye not be confused with a female fox. The only problem is from now on when I read a recipe calling for 0.25 grams of basic Fuchsin, I will see 0.25 grams of Basic vixen. ;) Rena Fail -----Original Message----- From: John Kiernan [mailto:jkiernan@uwo.ca] Sent: Wednesday, September 29, 2004 11:29 PM To: Rena; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Red chromogens [alkaline phosphatase labels] Dear Rena Fail, Red alkaline phosphatase product. Methods giving permanently mountable red products have been available for many years. Davis & Ornstein (1959) introduced hexazotized pararosaniline as a trifunctional trapping agent for naphthols released by hydrolysis of naphthol phosphates. Pararosaniline (CI 42500; CI Basic red 9) is one of the four components of basic fuchsine and is commercially available in fairly pure form. New fuchsine (CI 42520; CI Basic violet 2), which is closely similar to pararosaniline and also available in pure form (Horobin, 2002); It is currently preferred for making the hexazotized reagent. Kits are sold, but the substrate solution is easily made in the lab. In a simple procedure published at the IHCWorld.com (2004) web site (see below for its URL), the working mixture is prepared from four stock solutions, which are stored at 4 C. Solution D is warmed to room temperature before using. The amount of levamisole (inhibitor of endogenous tissue alkaline phosphatase) is clearly stated. The working solution is made immediately before using. Mix 10 drops each of A and B, then add 10 drops of solution C followed by 10 Kits are sold, but the substrate solution is easily made in the lab. In a simple procedure published at the IHCWorld.com (2004) web site, the working mixture is prepared from four stock solutions, which are stored at 4 C. Solution D is warmed to room temperature before using. I've annotated the instructions from IHCworld.com to simplify the weighing and measuring. A. New fuchsine (CI 42520), 0.2% in 2M HCl. (Conc. HCl is 10 or 12M) B. Sodium nitrite (NaNO2), 0.4% and levamisole,(= [-]-tetramisole hydrochloride) 0.48% in water. C. Naphthol AS-BI phosphate, 1% in 100% dimethylformamide. D. 0.05 M Tris-HCl buffer, pH 8.7. The working solution is made immediately before using. Mix 10 drops each of A and B, then add 10 drops of solution C followed by 10ml of the buffer (D). A drop is assumed to be 0.05 ml. Sections are incubated in the working solution for 10-20 minutes at room temperature. They are then washed counterstained as desired, dehydrated, cleared and coverslipped. Lojda et al (1979, p. 74-75) warn that the working solution may be red or brown from colored impurities in new fuchsine or pararosaniline. Such solutions cause excessive yellow background staining. The yellow/brown impurities are the same ones that can make Schiff's reagent yellow (removable from Schiff with activated charcoal). Pararosaniline certified by the Biological Stain Commission is suitable for making alkaline phosphatase substrate mixtures because it is required to be substantially free of brown and yellow materials (Penney et al., 2002). Probably any batch of basic fuchsine certified by the Biological Stain Commission will be OK for making the IHCWorld.com alkaline phosphatase substrate solution, because each of the four possible components of basic fuchsine has three diazotizable amino groups and their molecular weights are all quite close. I don't know of any published study that establishes this, so you're probably safest to go with either certified pararosaniline or with new fuchsine from a reputable dealer. References. (Please check them out if you can. The 2nd is easy enough!) Davis & Ornstein (1959) J. Histochem. Cytochem. 7:297-298. IHCWorld.com (2004) New fuchsin alkaline phosphatase substrate solution. http://www.ihcworld.com/_protocols/chromogen_substrates/AP_new_fuchsin_r ed.htm Horobin (2002) Chapter 14 in Conn's Biological Stains, 10th ed. Oxford:BIOS. Lojda et al (1979) Enzyme Histochemistry. Berlin: Springer. Penney et al. (2002) Biotech. Histochem. 77:237-275. Finally, please note that the correct spelling is fuchsine, not fuchsin, despite anything you might read in a catalog or on a bottle label. This is not a matter of American, British and German spellings. (Fuchsin is German for vixen; the dyes are, I think, named from their colours being similar to those of Fuchsia flowers rather than foxes.) Look in Websters, the Oxford, etc. It's fuchsine with an e. An -ine ending of an informal name of a chemical indicates that it is a base, often an amine. The -in ending occurs with compounds that are not bases, such as dextrin, eosin etc. John Kiernan London, Canada ------------------------------------------------------------------ Rena wrote: > > I have been having some trouble with some Abs stained with New Fuchsin > and permanent red. Recently I was asked if I were using Levamisole in > the permanent red and if so how much? I was told too much Levamisole in > permanent red would result in no staining. I have used 1 drop per ml for > both New Fuchsin and permanent red, which is the appropriate amount. We > have run both DABS and either permanent red or New Fuchsin with the > same AB from the same vial with vastly different results. Any ideas? > Rena Fail > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Thu Sep 30 04:00:12 2004 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Kluver-Barrier Stain References: Message-ID: <008101c4a6cb$e60b7920$f028d445@domainnotset.invalid> It's a Luxol fast blue LFB) with a cresyl echt violet (CEV) (also known as cresyl violet acetate) counterstain 0 LFB-CEV. I'll send you our procedure from work. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Elby, Lynette L" To: Sent: Wednesday, September 29, 2004 9:37 AM Subject: [Histonet] Kluver-Barrier Stain Does anyone have the procedure for this stain? If not, can you direct me to where I could find it? Thank in advance..... Lynette Elby Histology Supervisor Lancaster General Hospital 555 North Duke Street Lancaster, PA 17604 (717)544-5065 (717)544-5138 fax (717)305-6144 pager _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From psanquin <@t> lugo.usc.es Thu Sep 30 04:42:35 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Sucrose cryoprection In-Reply-To: <6.0.0.22.1.20040929120441.01af8d80@gemini.msu.montana.edu> References: <3.0.6.32.20040929191400.007f7ae0@pop.lugo.usc.es> <3.0.6.32.20040929191400.007f7ae0@pop.lugo.usc.es> Message-ID: <3.0.6.32.20040930114235.0080f100@pop.lugo.usc.es> Thank for your help Gayle, I am sorry my mail was not detailed enough. Yes, I am doing frozen sections with a freezing microtome. The animals are perfused with Paraformaldehyde 4% and postfixed for 48 hours in the same fixative. Then they are transfered to Phosphate buffer. I have problems when cutting the tissue. The sections (60 microns thick) are apparently nice, but just after putting them into the buffer they do not stay flat and they fall to pieces after gentle touching with the paintbrush. I have seen that after sinking in sucrose the samples show a gelatin-like appearance. They seem swollen and they are transparent. Could be this related to a poor fixation? For immunohistochemistry it is not recommended posfixation times greater tan 24 hours, but maybe postnatal or fetal material need longer postfixation time. Thanks for your interest. Pablo >We need more details on exactly what you are doing?????? and what the >problems are? Are you using a cryostat? > >At 11:14 AM 9/29/2004, you wrote: >>Dear listers, >> >>I am cutting at the sliding microtome fetal and early postantal mice >>brains. Could somebody tell me if is it normal that after sucrose >>cryoprection the pieces of tissue show a gelatin-like appearance? I have >>lot of problems cutting this tissue. >> >>Thanks in advance >> >>Pablo >> >> >> >> >>_______________________________________________ >>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 gu.lang <@t> gmx.at Thu Sep 30 04:40:10 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Red chromogens [alkaline phosphatase labels] References: <000001c4a68d$03c24780$c211a6a5@renad4yk9b8abe> <415B7D83.64C14C0A@uwo.ca> Message-ID: <001c01c4a6d1$7acdf470$eeeea8c0@server> Dear John, I think the German spelling is Fuchsin (without e). Please look at www.chroma.de at their catalogue. Your vixen is German "F?chsin" (fuechsin). greetings from Austria Gudrun ----- Original Message ----- From: "John Kiernan" To: "Rena" ; Sent: Thursday, September 30, 2004 5:29 AM Subject: Re: [Histonet] Red chromogens [alkaline phosphatase labels] Dear Rena Fail, Red alkaline phosphatase product. Methods giving permanently mountable red products have been available for many years. Davis & Ornstein (1959) introduced hexazotized pararosaniline as a trifunctional trapping agent for naphthols released by hydrolysis of naphthol phosphates. Pararosaniline (CI 42500; CI Basic red 9) is one of the four components of basic fuchsine and is commercially available in fairly pure form. New fuchsine (CI 42520; CI Basic violet 2), which is closely similar to pararosaniline and also available in pure form (Horobin, 2002); It is currently preferred for making the hexazotized reagent. Kits are sold, but the substrate solution is easily made in the lab. In a simple procedure published at the IHCWorld.com (2004) web site (see below for its URL), the working mixture is prepared from four stock solutions, which are stored at 4 C. Solution D is warmed to room temperature before using. The amount of levamisole (inhibitor of endogenous tissue alkaline phosphatase) is clearly stated. The working solution is made immediately before using. Mix 10 drops each of A and B, then add 10 drops of solution C followed by 10 Kits are sold, but the substrate solution is easily made in the lab. In a simple procedure published at the IHCWorld.com (2004) web site, the working mixture is prepared from four stock solutions, which are stored at 4 C. Solution D is warmed to room temperature before using. I've annotated the instructions from IHCworld.com to simplify the weighing and measuring. A. New fuchsine (CI 42520), 0.2% in 2M HCl. (Conc. HCl is 10 or 12M) B. Sodium nitrite (NaNO2), 0.4% and levamisole,(= [-]-tetramisole hydrochloride) 0.48% in water. C. Naphthol AS-BI phosphate, 1% in 100% dimethylformamide. D. 0.05 M Tris-HCl buffer, pH 8.7. The working solution is made immediately before using. Mix 10 drops each of A and B, then add 10 drops of solution C followed by 10ml of the buffer (D). A drop is assumed to be 0.05 ml. Sections are incubated in the working solution for 10-20 minutes at room temperature. They are then washed counterstained as desired, dehydrated, cleared and coverslipped. Lojda et al (1979, p. 74-75) warn that the working solution may be red or brown from colored impurities in new fuchsine or pararosaniline. Such solutions cause excessive yellow background staining. The yellow/brown impurities are the same ones that can make Schiff's reagent yellow (removable from Schiff with activated charcoal). Pararosaniline certified by the Biological Stain Commission is suitable for making alkaline phosphatase substrate mixtures because it is required to be substantially free of brown and yellow materials (Penney et al., 2002). Probably any batch of basic fuchsine certified by the Biological Stain Commission will be OK for making the IHCWorld.com alkaline phosphatase substrate solution, because each of the four possible components of basic fuchsine has three diazotizable amino groups and their molecular weights are all quite close. I don't know of any published study that establishes this, so you're probably safest to go with either certified pararosaniline or with new fuchsine from a reputable dealer. References. (Please check them out if you can. The 2nd is easy enough!) Davis & Ornstein (1959) J. Histochem. Cytochem. 7:297-298. IHCWorld.com (2004) New fuchsin alkaline phosphatase substrate solution. http://www.ihcworld.com/_protocols/chromogen_substrates/AP_new_fuchsin_red.htm Horobin (2002) Chapter 14 in Conn's Biological Stains, 10th ed. Oxford:BIOS. Lojda et al (1979) Enzyme Histochemistry. Berlin: Springer. Penney et al. (2002) Biotech. Histochem. 77:237-275. Finally, please note that the correct spelling is fuchsine, not fuchsin, despite anything you might read in a catalog or on a bottle label. This is not a matter of American, British and German spellings. (Fuchsin is German for vixen; the dyes are, I think, named from their colours being similar to those of Fuchsia flowers rather than foxes.) Look in Websters, the Oxford, etc. It's fuchsine with an e. An -ine ending of an informal name of a chemical indicates that it is a base, often an amine. The -in ending occurs with compounds that are not bases, such as dextrin, eosin etc. John Kiernan London, Canada ------------------------------------------------------------------ Rena wrote: > > I have been having some trouble with some Abs stained with New Fuchsin > and permanent red. Recently I was asked if I were using Levamisole in > the permanent red and if so how much? I was told too much Levamisole in > permanent red would result in no staining. I have used 1 drop per ml for > both New Fuchsin and permanent red, which is the appropriate amount. We > have run both DABS and either permanent red or New Fuchsin with the > same AB from the same vial with vastly different results. Any ideas? > Rena Fail > _______________________________________________ > Histonet mailing list > Histonet@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 David.Muskett <@t> RLC.NHS.UK Thu Sep 30 05:35:08 2004 From: David.Muskett <@t> RLC.NHS.UK (Muskett David) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Oxidising Agents in Haematoxylin[Scanned] Message-ID: <6AB79F0BA7C4914BA8DD8F5E0C21B97513BF37@AHEXMAIL01.xalderhey.com> Dear All I am inquiring about the use of differing oxidising agents in histology. I am routinely using Sodium Iodate as the oxidising agent in Ehrlich's haematoxylin. Can the sodium iodate be substituted by Potassium Iodate? Is there a difference in their oxidising properties? David David Muskett Chief Biomedical Scientist, Histology Royal Liverpool Children's NHS Trust Eaton Road Liverpool L12 2AP Telephone (0151) 293 3656 Fax (0151) 293 3617 From tbergeron <@t> criver.com Thu Sep 30 05:46:47 2004 From: tbergeron <@t> criver.com (tbergeron@criver.com) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] multi-cassette search In-Reply-To: Message-ID: Hi folks, We are currently using Evergreen multi-compartment cassettes for our Rodent GI tract samples, but are having issues with the lightness of the writing. It doesn't matter which brand of marker we use the writing comes out either very light, or washes partly off during processing. My assumption is it has to do with the type of plastic, which is a hard plastic similar to that used in the Sakura cassettes. So I am in search of a brand that is made with a softer plastic, similar to that used to make the Surgipath cassettes. Hopefully that makes sense. THANKS Tracy E. Bergeron, BS, HT(ASCP) Histotechician Charles River Laboratories Wilmington, MA 978-658-6000 x-1229 From rainbowmarie <@t> yahoo.com Thu Sep 30 05:57:34 2004 From: rainbowmarie <@t> yahoo.com (Mary Guerrero) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] UNSUSCRIBE Message-ID: <20040930105734.14681.qmail@web52710.mail.yahoo.com> Mary __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From c.m.vanderloos <@t> amc.uva.nl Thu Sep 30 06:27:42 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] RE: Red chromogens Message-ID: <1a7c2a1a8872.1a88721a7c2a@amc.uva.nl> Dear Rena, Too much levamisole cannot result in no staining. If you have used one drop (from the Dako concentrate??) per ml of substrate that should be OK (0.5 mM levamisole is considered as optimal). Too much of levamisole may result in somewhat weaker staining ultimately as levamisole inhibits the activity of all AP isoenzymes except the one from intestinal tissue. Your AP label is isolated from calf intestinal tissue. I can strongly recommend you to test the new Dako Permanent Red substrate (if already on the market). One suggestion: if you are a PBS user the phosphate ions from that buffer inhibit AP activity dramatically. Change to TBS or insert 2-3 washing steps with a non-phosphate containing buffer between your final PBS washing step and AP visualization. Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Rena Date Wed, 29 Sep 2004 21:30:04 -0400 To histonet@lists.utsouthwestern.edu Subject [Histonet] Red chromogens I have been having some trouble with some Abs stained with New Fuchsin and permanent red. Recently I was asked if I were using Levamisole in the permanent red and if so how much? I was told too much Levamisole in permanent red would result in no staining. I have used 1 drop per ml for both New Fuchsin and permanent red, which is the appropriate amount. We have run both DABS and either permanent red or New Fuchsin with the same AB from the same vial with vastly different results. Any ideas? Rena Fail From gu.lang <@t> gmx.at Thu Sep 30 06:36:09 2004 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Oxidising Agents in Haematoxylin[Scanned] References: <6AB79F0BA7C4914BA8DD8F5E0C21B97513BF37@AHEXMAIL01.xalderhey.com> Message-ID: <000e01c4a6e1$ae7cbf80$eeeea8c0@server> We use Potassium Iodate in Mayers Hematoxylin. Gudrun ----- Original Message ----- From: "Muskett David" To: Sent: Thursday, September 30, 2004 12:35 PM Subject: [Histonet] Oxidising Agents in Haematoxylin[Scanned] Dear All I am inquiring about the use of differing oxidising agents in histology. I am routinely using Sodium Iodate as the oxidising agent in Ehrlich's haematoxylin. Can the sodium iodate be substituted by Potassium Iodate? Is there a difference in their oxidising properties? David David Muskett Chief Biomedical Scientist, Histology Royal Liverpool Children's NHS Trust Eaton Road Liverpool L12 2AP Telephone (0151) 293 3656 Fax (0151) 293 3617 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AFeatherstone <@t> KaleidaHealth.Org Thu Sep 30 06:37:30 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Mast Cell Stain other than Toluidine Blue Message-ID: Does anyone have a simple Mast Cell Stain other than Toluidine Blue? Annette Featherstone HT/MLT -----Original Message----- From: hymclab [mailto:hymclab@hyhc.com] Sent: Wednesday, September 29, 2004 15:23 To: 'Andrea Grantham'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Bouins substitute in Trichrome staining PLEASE POST -----Original Message----- From: Andrea Grantham [mailto:algranth@u.arizona.edu] Sent: Wednesday, September 29, 2004 10:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bouins substitute in Trichrome staining I attended a workshop at NSH last week on connective tissue staining and one of the attendees said they were using citrate buffer instead of bouins as a mordant. I couldn't catch up with her after the workshop to ask about the protocol. Is anybody out in histoland doing this? Would you mind sharing your protocol if you are? 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 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 contact <@t> excaliburpathology.com Thu Sep 30 06:43:52 2004 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] multi-cassette search Message-ID: <20040930114352.84514.qmail@web50306.mail.yahoo.com> Hello Tracy, I still use pencil on my cassettes. I have tried all kinds of markers, but good old #2 lead works best, unless you can get a machine that heat stamps the # on the cassette. FYI NASA researchers have worked for years trying to improve on a pen that writes in the weightlessness of space, the Russians use pencils. LOL Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From mprice26 <@t> juno.com Thu Sep 30 08:16:51 2004 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] RE:Oil & Grease for the Leitz 1512 Message-ID: <20040930.061708.512.646565@webmail28.nyc.untd.com> Hi Everyone, I need to know where to order the oil and grease for the Leitz 1512 Microtome. Thank you. Marsha Price ________________________________________________________________ Get your name as your email address. Includes spam protection, 1GB storage, no ads and more Only $1.99/ month - visit http://www.mysite.com/name today! From Don.Birgerson <@t> leica-microsystems.com Thu Sep 30 08:57:20 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] RE:Oil & Grease for the Leitz 1512 Message-ID: Hi Marsha, Below are the "Leica" numbers for the old Leitz part numbers. Available from Leica Microsystems or their specimen prep dealers. Oil No. 601 50ml 14033621783 Leitz #11150000200116 Grease No.411 (white) 8/96 now 14033625075 Leitz #11150000200124 Grease No.410 (brown) 8/96 now 14033624601 Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "mprice26@juno.com" montana.edu Thu Sep 30 09:35:52 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Red chromogens In-Reply-To: <000001c4a68d$03c24780$c211a6a5@renad4yk9b8abe> References: <000001c4a68d$03c24780$c211a6a5@renad4yk9b8abe> Message-ID: <6.0.0.22.1.20040930082919.01b01148@gemini.msu.montana.edu> A talk with DAKO technical services can also help you with their chromogens. There is also a endog alk phos quenching solution from KPL, called Universal Blocker, and is used in very beginning of the staining protocol. It is ready to use and very nice, short, and very convenient. If you use this, levamisole is not needed (added to chromogen substrate which with ready to use chromogens, always worried me as a dilution factor) - in fact we tried the universal block in conjunction with levamisole to see effects and noticed lessening of red color or signal, not what we wanted. So you would have to use one or the other. I suggest you try the KPL blocker - If you are running your primary antibody at the same concentration for all these chromogens, you will notice a difference. This is something Chris van der Loos teaches in his multiple staining workshop but still applicable to single IHC work - about the efficiency of chromogens compared to antibody titer. With DAB you would have less conc primary than with new fuchsin, although Permanent red is approaching DAB for efficiency, some would call it sensitivity. At 07:30 PM 9/29/2004, you wrote: >I have been having some trouble with some Abs stained with New Fuchsin >and permanent red. Recently I was asked if I were using Levamisole in >the permanent red and if so how much? I was told too much Levamisole in >permanent red would result in no staining. I have used 1 drop per ml for >both New Fuchsin and permanent red, which is the appropriate amount. We >have run both DABS and either permanent red or New Fuchsin with the >same AB from the same vial with vastly different results. Any ideas? >Rena Fail >_______________________________________________ >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 cwscouten <@t> myneurolab.com Thu Sep 30 09:38:27 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Sucrose cryoprection Message-ID: It does sound like the tissue is not fixed, although 48 hours in Paraformaldehyde should be sufficient even without perfusion. Is the solution fully formed? Did any white powder remain in the bottom? It seems you are not getting good fixative. Did you prewash with saline or sucrose, or just perfuse? Blood can block many capillaries and block a through perfusion. 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 Pablo S?nchez Quinteiro Sent: Thursday, September 30, 2004 4:43 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sucrose cryoprection Thank for your help Gayle, I am sorry my mail was not detailed enough. Yes, I am doing frozen sections with a freezing microtome. The animals are perfused with Paraformaldehyde 4% and postfixed for 48 hours in the same fixative. Then they are transfered to Phosphate buffer. I have problems when cutting the tissue. The sections (60 microns thick) are apparently nice, but just after putting them into the buffer they do not stay flat and they fall to pieces after gentle touching with the paintbrush. I have seen that after sinking in sucrose the samples show a gelatin-like appearance. They seem swollen and they are transparent. Could be this related to a poor fixation? For immunohistochemistry it is not recommended posfixation times greater tan 24 hours, but maybe postnatal or fetal material need longer postfixation time. Thanks for your interest. Pablo >We need more details on exactly what you are doing?????? and what the >problems are? Are you using a cryostat? > >At 11:14 AM 9/29/2004, you wrote: >>Dear listers, >> >>I am cutting at the sliding microtome fetal and early postantal mice >>brains. Could somebody tell me if is it normal that after sucrose >>cryoprection the pieces of tissue show a gelatin-like appearance? I >>have lot of problems cutting this tissue. >> >>Thanks in advance >> >>Pablo >> >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From syedab <@t> totalise.co.uk Thu Sep 30 09:40:20 2004 From: syedab <@t> totalise.co.uk (Anila Syed) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Sucrose cryoprection References: <3.0.6.32.20040929191400.007f7ae0@pop.lugo.usc.es><3.0.6.32.20040929191400.007f7ae0@pop.lugo.usc.es> <3.0.6.32.20040930114235.0080f100@pop.lugo.usc.es> Message-ID: <002301c4a6fb$695b26c0$14c401a3@clneuro.ox.ac.uk> I used to put my tissue in gradually increasing concs of sucrose for up to 1 week *before* cutting them. But my samples were whole human brainstems, so your perfused animals may require less time. I used to try to get 50 micron thick sections of brainstem. Hope that helps? Anila ---- Original Message ----- From: "Pablo S?nchez Quinteiro" To: Sent: Thursday, September 30, 2004 10:42 AM Subject: Re: [Histonet] Sucrose cryoprection > Thank for your help Gayle, > > I am sorry my mail was not detailed enough. > > Yes, I am doing frozen sections with a freezing microtome. The animals are > perfused with Paraformaldehyde 4% and postfixed for 48 hours in the same > fixative. Then they are transfered to Phosphate buffer. > > I have problems when cutting the tissue. The sections (60 microns thick) > are apparently nice, but just after putting them into the buffer they do > not stay flat and they fall to pieces after gentle touching with the > paintbrush. > > I have seen that after sinking in sucrose the samples show a gelatin-like > appearance. They seem > swollen and they are transparent. Could be this related to a poor fixation? > > For immunohistochemistry it is not recommended posfixation times greater > tan 24 hours, but maybe postnatal or fetal material need longer > postfixation time. > > Thanks for your interest. > > Pablo > > >We need more details on exactly what you are doing?????? and what the > >problems are? Are you using a cryostat? > > > >At 11:14 AM 9/29/2004, you wrote: > >>Dear listers, > >> > >>I am cutting at the sliding microtome fetal and early postantal mice > >>brains. Could somebody tell me if is it normal that after sucrose > >>cryoprection the pieces of tissue show a gelatin-like appearance? I have > >>lot of problems cutting this tissue. > >> > >>Thanks in advance > >> > >>Pablo > >> > >> > >> > >> > >>_______________________________________________ > >>Histonet mailing list > >>Histonet@lists.utsouthwestern.edu > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >Gayle Callis > >MT,HT,HTL(ASCP) > >Research Histopathology Supervisor > >Veterinary Molecular Biology > >Montana State University - Bozeman > >PO Box 173610 > >Bozeman MT 59717-3610 > >406 994-6367 (lab with voice mail) > >406 994-4303 (FAX) > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.770 / Virus Database: 517 - Release Date: 27/09/04 From gcallis <@t> montana.edu Thu Sep 30 09:48:58 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Mast Cell Stain other than Toluidine Blue In-Reply-To: References: Message-ID: <6.0.0.22.1.20040930084605.01b136a8@gemini.msu.montana.edu> No, but I have two methods that are very simple, both are T blue. Churkian Schenk is my favorite, but have also used a method nicely passed on to me by Liz Chlipala out of Frieda Carson's book, a pH 4.3 T blue that takes 3 to 5 minutes, rinse with water, air dry the section and mount. This one is really simple and fast. If you want, I can forward both to you privately. At 05:37 AM 9/30/2004, you wrote: >Does anyone have a simple Mast Cell Stain other than Toluidine Blue? >Annette Featherstone HT/MLT >-----Original Message----- >From: hymclab [mailto:hymclab@hyhc.com] >Sent: Wednesday, September 29, 2004 15:23 >To: 'Andrea Grantham'; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Bouins substitute in Trichrome staining > > >PLEASE POST > >-----Original Message----- >From: Andrea Grantham [mailto:algranth@u.arizona.edu] >Sent: Wednesday, September 29, 2004 10:38 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Bouins substitute in Trichrome staining > > >I attended a workshop at NSH last week on connective tissue staining and >one of the attendees said they were using citrate buffer instead of bouins >as a mordant. I couldn't catch up with her after the workshop to ask about >the protocol. Is anybody out in histoland doing this? Would you mind >sharing your protocol if you are? >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 > >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 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 mward <@t> wfubmc.edu Thu Sep 30 11:12:09 2004 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] FXIII antibody Message-ID: <61135F0455D33347B5AAE209B903A304076A4F6A@EXCHVS2.medctr.ad.wfubmc.edu> I am looking for a vendor for the FXIIIa antibody. I tried DAKO, Calbiochem and Biogenex and no one carries it. Any help will be appreciated. Martha Ward Molecular Diagnostics Lab Wake Forest University Baptist Medical Center From JGordon <@t> cellmarque.com Thu Sep 30 11:29:22 2004 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] FXIII antibody Message-ID: Based on the vendors that you have inquired upon that do not offer it, I'm not sure which companies carry (or carried) Factor 13a, but we do have it at Cell Marque, if that is an option for you. It is available as both a concentrate and a ready-to-use. Jeff Gordon Cell Marque Corp. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Martha Ward Sent: Thursday, September 30, 2004 11:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] FXIII antibody I am looking for a vendor for the FXIIIa antibody. I tried DAKO, Calbiochem and Biogenex and no one carries it. Any help will be appreciated. Martha Ward Molecular Diagnostics Lab Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From m.wei <@t> biogenex.com Thu Sep 30 11:31:57 2004 From: m.wei <@t> biogenex.com (May Wei) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] FXIII antibody Message-ID: Dear Martha, BioGenex (1-800-421-4149) has released this Factor XIIIa antibody recently. It is monoclonal with clone [E980.1]. The RTU catalog number is AM337-5M. The concentrated format catalog number is MU337-UC. May Wei, M. Med., MBA International Account Manager BioGenex Laboratories 4600 Norris Canyon Road San Ramon, CA 94583 Tel: (925) 543-1350 Fax: (925) 866-2525 Email: m.wei@biogenex.com -----Original Message----- From: Martha Ward [mailto:mward@wfubmc.edu] Sent: Thursday, September 30, 2004 9:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] FXIII antibody I am looking for a vendor for the FXIIIa antibody. I tried DAKO, Calbiochem and Biogenex and no one carries it. Any help will be appreciated. Martha Ward Molecular Diagnostics Lab Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From themagoos <@t> rushmore.com Thu Sep 30 11:33:44 2004 From: themagoos <@t> rushmore.com (Jason and Heather) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] FXIII antibody Message-ID: <001e01c4a70b$427c7cb0$b3fc9fd1@magoo> We use a mouse antibody from Cell Marque. We have had good luck with it. Jason McGoughHT(ASCP) Histology Clinical Lab of the Black Hills From japoteete <@t> saintfrancis.com Thu Sep 30 11:32:19 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] FXIII antibody Message-ID: We use CellMarque CMC780 Factor XIIIa with good results. Their number is 1-800-665-7284. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com > -----Original Message----- > From: Martha Ward [SMTP:mward@wfubmc.edu] > Sent: Thursday, September 30, 2004 11:12 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] FXIII antibody > > I am looking for a vendor for the FXIIIa antibody. I tried DAKO, > Calbiochem and Biogenex and no one carries it. Any help will be > appreciated. > > Martha Ward > Molecular Diagnostics Lab > Wake Forest University Baptist Medical Center > _______________________________________________ > 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 gcallis <@t> montana.edu Thu Sep 30 12:09:09 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Embedding centers - need feedback on new models to replace dying unit Message-ID: <6.0.0.22.1.20040930110520.01b032c0@gemini.msu.montana.edu> Dear All, I would like private feedback on embedding centers, new models, people are now using. My beloved Medim DDM 0065 is slowing dying an electrical death, although cryo and paraffin dispenser units still work! I realize embedding centers are pricey, but what isn't these days, please forward pros and cons to me on usage. Also, any feedback on who repairs these, maybe a used equipment dealer, would be appreciated. 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 sharon.osborn <@t> dnax.org Thu Sep 30 12:14:57 2004 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] RE: Mast cell staining Message-ID: <29B25753F6B1D51196110002A589D44401A17845@PALMSG30.us.schp.com> Annette Featherstone, Lee Luna's book has a beautiful mast cell stain using the T-blue and with eosin as a counter stain. It is Luna's Toluidine Blue Method for Mast Cells on Pages 311-312 of Luna: Histological Methods, Color Atlas, Special Stains, etc. If you wish to have the protocol, I can email you privately. 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 timothy_m_coskran <@t> groton.pfizer.com Thu Sep 30 12:19:56 2004 From: timothy_m_coskran <@t> groton.pfizer.com (Coskran, Timothy M) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Red Chromagens Message-ID: <75FBD3E83B69D711A9E600080261980C02BC9FA1@groexmb04bak.pfizer.com> A note about Permanent Red. The spec sheet recommends bring the reagents to room temperature before dissolving the tablet in the substrate buffer. In our hands this is a must. When we've forgotten to bring the reagents to room temp, and went ahead and dissolved the tablet and used the reagents, the staining was patchy or absent. We routinely pull the necessary amount of tablets and substrate buffer out of the fridge 30 minutes prior to the Dako Autostainer Batch step. The staining has been great. Tim Coskran Pfizer LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From DSpears <@t> agr.state.il.us Thu Sep 30 12:42:25 2004 From: DSpears <@t> agr.state.il.us (DSpears@agr.state.il.us) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Embedding centers - need feedback on new models to replace dying unit In-Reply-To: <6.0.0.22.1.20040930110520.01b032c0@gemini.msu.montana.edu> Message-ID: Gayle, We really love the Tissue Tek TEC - have never had any problems with it or the previous one that I now use solely for prion-infected tissue. I did see a cool one at NSH - I'm not sure but think it might have been TBS. It had an infrared sensor and so you don't need a footpedal or pressure switch - when you break the beam with the mold it just dispenses. Dana Spears Gayle Callis Sent by: histonet-bounces@lists.utsouthwestern.edu 09/30/2004 12:09 PM To Histonet@lists.utsouthwestern.edu cc Subject [Histonet] Embedding centers - need feedback on new models to replace dying unit Dear All, I would like private feedback on embedding centers, new models, people are now using. My beloved Medim DDM 0065 is slowing dying an electrical death, although cryo and paraffin dispenser units still work! I realize embedding centers are pricey, but what isn't these days, please forward pros and cons to me on usage. Also, any feedback on who repairs these, maybe a used equipment dealer, would be appreciated. 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) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Thu Sep 30 12:38:50 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] FXIII antibody Message-ID: Martha, We use Biocare Medical's polyclonal F13a with great results on our Ventana instruments. Their number is: (800)799-9499. 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: Martha Ward [mailto:mward@wfubmc.edu] Sent: Thursday, September 30, 2004 11:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] FXIII antibody I am looking for a vendor for the FXIIIa antibody. I tried DAKO, Calbiochem and Biogenex and no one carries it. Any help will be appreciated. Martha Ward Molecular Diagnostics Lab Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Julie.Sanders <@t> med.va.gov Thu Sep 30 12:40:07 2004 From: Julie.Sanders <@t> med.va.gov (Julie.Sanders@med.va.gov) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Factor XIIIa Message-ID: <457381D92B01BD44B21CF37CC02EBDFD28E965@vhacinexc2.v10.med.va.gov> We get our from Cell Marque and have good results on the Ventana Benchmark. Julie Julie Sanders,BA,HT(ASCP) Supervisor, Anatomic Pathology VAMC Cincinnati, Ohio -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Thursday, September 30, 2004 1:14 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 10, Issue 40 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: Red chromogens (C.M. van der Loos) 2. Re: Oxidising Agents in Haematoxylin[Scanned] (Gudrun Lang) 3. Mast Cell Stain other than Toluidine Blue (Featherstone, Annette) 4. multi-cassette search (Paula Pierce) 5. RE:Oil & Grease for the Leitz 1512 (mprice26@juno.com) 6. Re: RE:Oil & Grease for the Leitz 1512 (Don.Birgerson@leica-microsystems.com) 7. Re: Red chromogens (Gayle Callis) 8. RE: Sucrose cryoprection (Charles Scouten) 9. Re: Sucrose cryoprection (Anila Syed) 10. Re: Mast Cell Stain other than Toluidine Blue (Gayle Callis) 11. FXIII antibody (Martha Ward) 12. RE: FXIII antibody (Jeff Gordon) 13. RE: FXIII antibody (May Wei) 14. FXIII antibody (Jason and Heather) 15. RE: FXIII antibody (Poteete, Jacquie A.) ---------------------------------------------------------------------- Message: 1 Date: Thu, 30 Sep 2004 13:27:42 +0200 From: "C.M. van der Loos" Subject: [Histonet] RE: Red chromogens To: histonet@lists.utsouthwestern.edu Cc: RFail@Charleston.net Message-ID: <1a7c2a1a8872.1a88721a7c2a@amc.uva.nl> Content-Type: text/plain; charset=us-ascii Dear Rena, Too much levamisole cannot result in no staining. If you have used one drop (from the Dako concentrate??) per ml of substrate that should be OK (0.5 mM levamisole is considered as optimal). Too much of levamisole may result in somewhat weaker staining ultimately as levamisole inhibits the activity of all AP isoenzymes except the one from intestinal tissue. Your AP label is isolated from calf intestinal tissue. I can strongly recommend you to test the new Dako Permanent Red substrate (if already on the market). One suggestion: if you are a PBS user the phosphate ions from that buffer inhibit AP activity dramatically. Change to TBS or insert 2-3 washing steps with a non-phosphate containing buffer between your final PBS washing step and AP visualization. Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Rena Date Wed, 29 Sep 2004 21:30:04 -0400 To histonet@lists.utsouthwestern.edu Subject [Histonet] Red chromogens I have been having some trouble with some Abs stained with New Fuchsin and permanent red. Recently I was asked if I were using Levamisole in the permanent red and if so how much? I was told too much Levamisole in permanent red would result in no staining. I have used 1 drop per ml for both New Fuchsin and permanent red, which is the appropriate amount. We have run both DABS and either permanent red or New Fuchsin with the same AB from the same vial with vastly different results. Any ideas? Rena Fail ------------------------------ Message: 2 Date: Thu, 30 Sep 2004 13:36:09 +0200 From: "Gudrun Lang" Subject: Re: [Histonet] Oxidising Agents in Haematoxylin[Scanned] To: "Histonetliste" Message-ID: <000e01c4a6e1$ae7cbf80$eeeea8c0@server> Content-Type: text/plain; charset="iso-8859-1" We use Potassium Iodate in Mayers Hematoxylin. Gudrun ----- Original Message ----- From: "Muskett David" To: Sent: Thursday, September 30, 2004 12:35 PM Subject: [Histonet] Oxidising Agents in Haematoxylin[Scanned] Dear All I am inquiring about the use of differing oxidising agents in histology. I am routinely using Sodium Iodate as the oxidising agent in Ehrlich's haematoxylin. Can the sodium iodate be substituted by Potassium Iodate? Is there a difference in their oxidising properties? David David Muskett Chief Biomedical Scientist, Histology Royal Liverpool Children's NHS Trust Eaton Road Liverpool L12 2AP Telephone (0151) 293 3656 Fax (0151) 293 3617 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Thu, 30 Sep 2004 07:37:30 -0400 From: "Featherstone, Annette" Subject: [Histonet] Mast Cell Stain other than Toluidine Blue To: 'hymclab' , 'Andrea Grantham' , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Does anyone have a simple Mast Cell Stain other than Toluidine Blue? Annette Featherstone HT/MLT -----Original Message----- From: hymclab [mailto:hymclab@hyhc.com] Sent: Wednesday, September 29, 2004 15:23 To: 'Andrea Grantham'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Bouins substitute in Trichrome staining PLEASE POST -----Original Message----- From: Andrea Grantham [mailto:algranth@u.arizona.edu] Sent: Wednesday, September 29, 2004 10:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bouins substitute in Trichrome staining I attended a workshop at NSH last week on connective tissue staining and one of the attendees said they were using citrate buffer instead of bouins as a mordant. I couldn't catch up with her after the workshop to ask about the protocol. Is anybody out in histoland doing this? Would you mind sharing your protocol if you are? 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 CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. ------------------------------ Message: 4 Date: Thu, 30 Sep 2004 04:43:52 -0700 (PDT) From: Paula Pierce Subject: [Histonet] multi-cassette search To: histonet@lists.utsouthwestern.edu Message-ID: <20040930114352.84514.qmail@web50306.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hello Tracy, I still use pencil on my cassettes. I have tried all kinds of markers, but good old #2 lead works best, unless you can get a machine that heat stamps the # on the cassette. FYI NASA researchers have worked for years trying to improve on a pen that writes in the weightlessness of space, the Russians use pencils. LOL Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com ------------------------------ Message: 5 Date: Thu, 30 Sep 2004 13:16:51 GMT From: "mprice26@juno.com" Subject: [Histonet] RE:Oil & Grease for the Leitz 1512 To: histonet@lists.utsouthwestern.edu Message-ID: <20040930.061708.512.646565@webmail28.nyc.untd.com> Content-Type: text/plain Hi Everyone, I need to know where to order the oil and grease for the Leitz 1512 Microtome. Thank you. Marsha Price ________________________________________________________________ Get your name as your email address. Includes spam protection, 1GB storage, no ads and more Only $1.99/ month - visit http://www.mysite.com/name today! ------------------------------ Message: 6 Date: Thu, 30 Sep 2004 08:57:20 -0500 From: Don.Birgerson@leica-microsystems.com Subject: Re: [Histonet] RE:Oil & Grease for the Leitz 1512 To: "mprice26@juno.com" Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Hi Marsha, Below are the "Leica" numbers for the old Leitz part numbers. Available from Leica Microsystems or their specimen prep dealers. Oil No. 601 50ml 14033621783 Leitz #11150000200116 Grease No.411 (white) 8/96 now 14033625075 Leitz #11150000200124 Grease No.410 (brown) 8/96 now 14033624601 Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "mprice26@juno.com" Subject: Re: [Histonet] Red chromogens To: Rena , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20040930082919.01b01148@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed A talk with DAKO technical services can also help you with their chromogens. There is also a endog alk phos quenching solution from KPL, called Universal Blocker, and is used in very beginning of the staining protocol. It is ready to use and very nice, short, and very convenient. If you use this, levamisole is not needed (added to chromogen substrate which with ready to use chromogens, always worried me as a dilution factor) - in fact we tried the universal block in conjunction with levamisole to see effects and noticed lessening of red color or signal, not what we wanted. So you would have to use one or the other. I suggest you try the KPL blocker - If you are running your primary antibody at the same concentration for all these chromogens, you will notice a difference. This is something Chris van der Loos teaches in his multiple staining workshop but still applicable to single IHC work - about the efficiency of chromogens compared to antibody titer. With DAB you would have less conc primary than with new fuchsin, although Permanent red is approaching DAB for efficiency, some would call it sensitivity. At 07:30 PM 9/29/2004, you wrote: >I have been having some trouble with some Abs stained with New Fuchsin >and permanent red. Recently I was asked if I were using Levamisole in >the permanent red and if so how much? I was told too much Levamisole in >permanent red would result in no staining. I have used 1 drop per ml for >both New Fuchsin and permanent red, which is the appropriate amount. We >have run both DABS and either permanent red or New Fuchsin with the >same AB from the same vial with vastly different results. Any ideas? >Rena Fail >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 8 Date: Thu, 30 Sep 2004 09:38:27 -0500 From: "Charles Scouten" Subject: RE: [Histonet] Sucrose cryoprection To: Pablo S?nchez Quinteiro , Message-ID: Content-Type: text/plain; charset="iso-8859-1" It does sound like the tissue is not fixed, although 48 hours in Paraformaldehyde should be sufficient even without perfusion. Is the solution fully formed? Did any white powder remain in the bottom? It seems you are not getting good fixative. Did you prewash with saline or sucrose, or just perfuse? Blood can block many capillaries and block a through perfusion. 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 Pablo S?nchez Quinteiro Sent: Thursday, September 30, 2004 4:43 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Sucrose cryoprection Thank for your help Gayle, I am sorry my mail was not detailed enough. Yes, I am doing frozen sections with a freezing microtome. The animals are perfused with Paraformaldehyde 4% and postfixed for 48 hours in the same fixative. Then they are transfered to Phosphate buffer. I have problems when cutting the tissue. The sections (60 microns thick) are apparently nice, but just after putting them into the buffer they do not stay flat and they fall to pieces after gentle touching with the paintbrush. I have seen that after sinking in sucrose the samples show a gelatin-like appearance. They seem swollen and they are transparent. Could be this related to a poor fixation? For immunohistochemistry it is not recommended posfixation times greater tan 24 hours, but maybe postnatal or fetal material need longer postfixation time. Thanks for your interest. Pablo >We need more details on exactly what you are doing?????? and what the >problems are? Are you using a cryostat? > >At 11:14 AM 9/29/2004, you wrote: >>Dear listers, >> >>I am cutting at the sliding microtome fetal and early postantal mice >>brains. Could somebody tell me if is it normal that after sucrose >>cryoprection the pieces of tissue show a gelatin-like appearance? I >>have lot of problems cutting this tissue. >> >>Thanks in advance >> >>Pablo >> >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Thu, 30 Sep 2004 15:40:20 +0100 From: "Anila Syed" Subject: Re: [Histonet] Sucrose cryoprection To: , Pablo S?nchez Quinteiro Message-ID: <002301c4a6fb$695b26c0$14c401a3@clneuro.ox.ac.uk> Content-Type: text/plain; charset="iso-8859-1" I used to put my tissue in gradually increasing concs of sucrose for up to 1 week *before* cutting them. But my samples were whole human brainstems, so your perfused animals may require less time. I used to try to get 50 micron thick sections of brainstem. Hope that helps? Anila ---- Original Message ----- From: "Pablo S?nchez Quinteiro" To: Sent: Thursday, September 30, 2004 10:42 AM Subject: Re: [Histonet] Sucrose cryoprection > Thank for your help Gayle, > > I am sorry my mail was not detailed enough. > > Yes, I am doing frozen sections with a freezing microtome. The animals are > perfused with Paraformaldehyde 4% and postfixed for 48 hours in the same > fixative. Then they are transfered to Phosphate buffer. > > I have problems when cutting the tissue. The sections (60 microns thick) > are apparently nice, but just after putting them into the buffer they do > not stay flat and they fall to pieces after gentle touching with the > paintbrush. > > I have seen that after sinking in sucrose the samples show a gelatin-like > appearance. They seem > swollen and they are transparent. Could be this related to a poor fixation? > > For immunohistochemistry it is not recommended posfixation times greater > tan 24 hours, but maybe postnatal or fetal material need longer > postfixation time. > > Thanks for your interest. > > Pablo > > >We need more details on exactly what you are doing?????? and what the > >problems are? Are you using a cryostat? > > > >At 11:14 AM 9/29/2004, you wrote: > >>Dear listers, > >> > >>I am cutting at the sliding microtome fetal and early postantal mice > >>brains. Could somebody tell me if is it normal that after sucrose > >>cryoprection the pieces of tissue show a gelatin-like appearance? I have > >>lot of problems cutting this tissue. > >> > >>Thanks in advance > >> > >>Pablo > >> > >> > >> > >> > >>_______________________________________________ > >>Histonet mailing list > >>Histonet@lists.utsouthwestern.edu > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >Gayle Callis > >MT,HT,HTL(ASCP) > >Research Histopathology Supervisor > >Veterinary Molecular Biology > >Montana State University - Bozeman > >PO Box 173610 > >Bozeman MT 59717-3610 > >406 994-6367 (lab with voice mail) > >406 994-4303 (FAX) > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.770 / Virus Database: 517 - Release Date: 27/09/04 ------------------------------ Message: 10 Date: Thu, 30 Sep 2004 08:48:58 -0600 From: Gayle Callis Subject: Re: [Histonet] Mast Cell Stain other than Toluidine Blue To: "Featherstone, Annette" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20040930084605.01b136a8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed No, but I have two methods that are very simple, both are T blue. Churkian Schenk is my favorite, but have also used a method nicely passed on to me by Liz Chlipala out of Frieda Carson's book, a pH 4.3 T blue that takes 3 to 5 minutes, rinse with water, air dry the section and mount. This one is really simple and fast. If you want, I can forward both to you privately. At 05:37 AM 9/30/2004, you wrote: >Does anyone have a simple Mast Cell Stain other than Toluidine Blue? >Annette Featherstone HT/MLT >-----Original Message----- >From: hymclab [mailto:hymclab@hyhc.com] >Sent: Wednesday, September 29, 2004 15:23 >To: 'Andrea Grantham'; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Bouins substitute in Trichrome staining > > >PLEASE POST > >-----Original Message----- >From: Andrea Grantham [mailto:algranth@u.arizona.edu] >Sent: Wednesday, September 29, 2004 10:38 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Bouins substitute in Trichrome staining > > >I attended a workshop at NSH last week on connective tissue staining and >one of the attendees said they were using citrate buffer instead of bouins >as a mordant. I couldn't catch up with her after the workshop to ask about >the protocol. Is anybody out in histoland doing this? Would you mind >sharing your protocol if you are? >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 > >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 Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 11 Date: Thu, 30 Sep 2004 12:12:09 -0400 From: "Martha Ward" Subject: [Histonet] FXIII antibody To: Message-ID: <61135F0455D33347B5AAE209B903A304076A4F6A@EXCHVS2.medctr.ad.wfubmc.edu> Content-Type: text/plain; charset="US-ASCII" I am looking for a vendor for the FXIIIa antibody. I tried DAKO, Calbiochem and Biogenex and no one carries it. Any help will be appreciated. Martha Ward Molecular Diagnostics Lab Wake Forest University Baptist Medical Center ------------------------------ Message: 12 Date: Thu, 30 Sep 2004 11:29:22 -0500 From: "Jeff Gordon" Subject: RE: [Histonet] FXIII antibody To: "Martha Ward" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Based on the vendors that you have inquired upon that do not offer it, I'm not sure which companies carry (or carried) Factor 13a, but we do have it at Cell Marque, if that is an option for you. It is available as both a concentrate and a ready-to-use. Jeff Gordon Cell Marque Corp. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Martha Ward Sent: Thursday, September 30, 2004 11:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] FXIII antibody I am looking for a vendor for the FXIIIa antibody. I tried DAKO, Calbiochem and Biogenex and no one carries it. Any help will be appreciated. Martha Ward Molecular Diagnostics Lab Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Thu, 30 Sep 2004 09:31:57 -0700 From: May Wei Subject: RE: [Histonet] FXIII antibody To: 'Martha Ward' Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Dear Martha, BioGenex (1-800-421-4149) has released this Factor XIIIa antibody recently. It is monoclonal with clone [E980.1]. The RTU catalog number is AM337-5M. The concentrated format catalog number is MU337-UC. May Wei, M. Med., MBA International Account Manager BioGenex Laboratories 4600 Norris Canyon Road San Ramon, CA 94583 Tel: (925) 543-1350 Fax: (925) 866-2525 Email: m.wei@biogenex.com -----Original Message----- From: Martha Ward [mailto:mward@wfubmc.edu] Sent: Thursday, September 30, 2004 9:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] FXIII antibody I am looking for a vendor for the FXIIIa antibody. I tried DAKO, Calbiochem and Biogenex and no one carries it. Any help will be appreciated. Martha Ward Molecular Diagnostics Lab Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Thu, 30 Sep 2004 10:33:44 -0600 From: "Jason and Heather" Subject: [Histonet] FXIII antibody To: Message-ID: <001e01c4a70b$427c7cb0$b3fc9fd1@magoo> Content-Type: text/plain; charset="iso-8859-1" We use a mouse antibody from Cell Marque. We have had good luck with it. Jason McGoughHT(ASCP) Histology Clinical Lab of the Black Hills ------------------------------ Message: 15 Date: Thu, 30 Sep 2004 11:32:19 -0500 From: "Poteete, Jacquie A." Subject: RE: [Histonet] FXIII antibody To: 'Martha Ward' , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain We use CellMarque CMC780 Factor XIIIa with good results. Their number is 1-800-665-7284. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com > -----Original Message----- > From: Martha Ward [SMTP:mward@wfubmc.edu] > Sent: Thursday, September 30, 2004 11:12 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] FXIII antibody > > I am looking for a vendor for the FXIIIa antibody. I tried DAKO, > Calbiochem and Biogenex and no one carries it. Any help will be > appreciated. > > Martha Ward > Molecular Diagnostics Lab > Wake Forest University Baptist Medical Center > _______________________________________________ > 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 End of Histonet Digest, Vol 10, Issue 40 **************************************** From gcallis <@t> montana.edu Thu Sep 30 12:48:17 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Histology equipment repairs Message-ID: <6.0.0.22.1.20040930114640.01aee700@gemini.msu.montana.edu> Dear All, I have contacted IMEB about repair, but also would like to know of another repair service, preferrably in the Western region for histology equipment repair, i.e. for an embedding center - just another option instead of purchasing used or new. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From froyer <@t> bitstream.net Thu Sep 30 13:53:10 2004 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Histology equipment repairs In-Reply-To: <6.0.0.22.1.20040930114640.01aee700@gemini.msu.montana.edu> References: <6.0.0.22.1.20040930114640.01aee700@gemini.msu.montana.edu> Message-ID: <415C5616.40205@bitstream.net> We SERVICE, REPAIR, and SELL new and used Refurbished Histology Equipment nation-wide. (and we're close to Montana) ;-) Located in the Twin Cities of Minneapolis/ St. Paul, MN I'll be glad to give you an estimate on the repair of your embedding center, or a quote on a refurbished unit. ~ Ford Ford M. Royer, MT(ASCP) (Formerly with Analytical Instruments, LLC) National Sales Manager Midwest Science Biocenter 6551 Jansen Ave. NE, Ste. 102 Albertville, MN 55301-4316 (800) 745-4869, Ext. 154 URL: Gayle Callis wrote: > Dear All, > > I have contacted IMEB about repair, but also would like to know of > another repair service, preferrably in the Western region for > histology equipment repair, i.e. for an embedding center - just > another option instead of purchasing used or new. > > 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 psanquin <@t> lugo.usc.es Thu Sep 30 14:17:34 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Sucrose cryoprection Message-ID: <3.0.6.32.20040930211734.007db4b0@pop.lugo.usc.es> Thanks to everybody who has made me on- and off-list useful and interesting hints. Now it seems clear to me that I have a problem with fixation. I'll try to be more careful; for example making prewash (I do not do that in order to simplify the complicated procedure of perfusing so tiny animals) or checking if there is powder of Paraformaldehyde in the botton (Some times a few grains remain in the bottom). I think that in addition to this the postfixation time must have the key role. By the way I would like to know your experience with nervous tissue in young animals (1-7 days old in mouse, for example). It is not easy to fix properly this postnatal animals. When I perfuse the paraformaldehyde in adult animals I can see the sad limbs movements which indicate that fixation is going well. In postnatal animals I never get this effect. Have you got any idea regarding this difference? Maybe the more water content in postnatal animals or perhaps -as cleverly a co-lister has suggested me- the more immature CNS system in postanatal animals. Thanks again Pablo X-Virus-Scanned: by usc.es X-MIME-Autoconverted: from quoted-printable to 8bit by secuslugo.lugo.usc.es id i8UEd9O26589 It does sound like the tissue is not fixed, although 48 hours in Paraformaldehyde should be sufficient even without perfusion. Is the solution fully formed? Did any white powder remain in the bottom? It seems you are not getting good fixative. Did you prewash with saline or sucrose, or just perfuse? Blood can block many capillaries and block a through perfusion. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com From sbreeden <@t> nmda.nmsu.edu Thu Sep 30 14:43:40 2004 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Oil & Grease for Leitz 1512 Message-ID: If you will contact Nick Klados at 303-841-5174, he will put you in touch with a supplier. Nick does maintenace on the Leitz 1512s. From kabrady <@t> genomichealth.com Thu Sep 30 16:24:27 2004 From: kabrady <@t> genomichealth.com (Kimberly Brady) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Traveling Histotech Message-ID: <5C9B539E7DD26C4AA41D4E27ACECBB910197BE94@jaguar.genomichealth.com> Does anyone out there have any contact info for traveling histotechs? I am interested in finding out how much it would cost to hire an HT/HTL (ASCP) for 1-3 days. Please email me directly to my email address if anyone has any information. Thanks, Kim The contents of this electronic message, including any attachments, are intended only for the use of the individual or entity to which they are addressed and may contain confidential information. If you are not the intended recipient, you are hereby notified that any use, dissemination, distribution, or copying of this message or any attachment is strictly prohibited. If you have received this transmission in error, please send an e-mail to postmaster@genomichealth.com and delete this message, along with any attachments, from your computer. From Kathy.Paton <@t> waitematadhb.govt.nz Thu Sep 30 16:45:44 2004 From: Kathy.Paton <@t> waitematadhb.govt.nz (Kathy Paton (WDHB)) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Mast Cell Stain other than Toluidine Blue Message-ID: My favorite for mast cell granules has to be the Csaba from Disbrey and Racks publication. It uses Alcian blue and safranin in Walpoles/HCL buffer pH 1.42. Not only does it stain mast cells beautifully but it will differentiate between young mast cell granules that are histamine dominant (blue) and mature mast cell granules that are heparin dominant (red) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Friday, 1 October 2004 02:49 To: Featherstone, Annette; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Mast Cell Stain other than Toluidine Blue No, but I have two methods that are very simple, both are T blue. Churkian Schenk is my favorite, but have also used a method nicely passed on to me by Liz Chlipala out of Frieda Carson's book, a pH 4.3 T blue that takes 3 to 5 minutes, rinse with water, air dry the section and mount. This one is really simple and fast. If you want, I can forward both to you privately. At 05:37 AM 9/30/2004, you wrote: >Does anyone have a simple Mast Cell Stain other than Toluidine Blue? >Annette Featherstone HT/MLT >-----Original Message----- >From: hymclab [mailto:hymclab@hyhc.com] >Sent: Wednesday, September 29, 2004 15:23 >To: 'Andrea Grantham'; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Bouins substitute in Trichrome staining > > >PLEASE POST > >-----Original Message----- >From: Andrea Grantham [mailto:algranth@u.arizona.edu] >Sent: Wednesday, September 29, 2004 10:38 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Bouins substitute in Trichrome staining > > >I attended a workshop at NSH last week on connective tissue staining >and one of the attendees said they were using citrate buffer instead of >bouins as a mordant. I couldn't catch up with her after the workshop to >ask about the protocol. Is anybody out in histoland doing this? Would >you mind sharing your protocol if you are? >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 > >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 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 ccrowder <@t> mail.vetmed.lsu.edu Thu Sep 30 16:50:29 2004 From: ccrowder <@t> mail.vetmed.lsu.edu (Cheryl Crowder) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Mast cell stain Message-ID: Hi - We do hundreds of stains for mast cells each year. The best we have found is either the T-blue, but we use pH 2.5 or a Jenner-Giemsa. The Giemsa will not only stain the mast cells but also bacteria and other structures. I can send our procedures is you want them. 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 jbaez <@t> interscopepath.com Thu Sep 30 17:00:04 2004 From: jbaez <@t> interscopepath.com (Baez, Janet) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] away Message-ID: <9E956D8FEB06C2408B08AC16498325E901399C@scopemx1.interscope.com> I will be away on vacation from October 6,2004 through November 1, 2004. Please do not forward e-mails to me. Thank you. From ynwang <@t> u.washington.edu Thu Sep 30 17:13:28 2004 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] sirius red Message-ID: Dear all, I have a question regarding sirius red staining. Background: We are looking at amino acid based polyurethane 'mesh' (fibrous structure) that are unseeded or seeded with porcine mitral valve cells. The seeded scaffolds were either loaded or unloaded. We are investigating the collagen production (we have used a pro-collagen assay as well as a hydroxyproline assay and have come up with positive results). In the past we have used Massons Trichrome to get some pretty pictures, however, even using the room temperature mordent (overnight) was not good for the polymer (biodegradable) so we have started to use sirius red staining (thank you to the histonet members for the protocols we found on the archives!) Sections: samples were embedded in OCT. 5um sections were taken to be stained. There is one sample that we see staining (reddish)under bright-field and polarized light (we are assuming this is collagen). For the majority of the seeded scaffolds we are seeing 'cell-like' red staining which does not show under polarized light (cytoplasm?). However, with the unseeded controls we see similar (although not really cell-like) staining which is predominently at/near the sample-OCT interface. Does anyone have any ideas as to what it would be? I was thinking it could possibly be trapped dye where the section has folded in slightly? The scaffold itself is staining slightly pink too. Thanks for your help Yak-Nam Wang Senior Fellow Department of Bioengineering University of Washington Seattle, WA 98195 From AnthonyH <@t> chw.edu.au Thu Sep 30 18:05:26 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Mast Cell Stain other than Toluidine Blue Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E0C3@simba.kids> The alcian blue at pH 1 is a good demonstration method for mast cells. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Friday, 1 October 2004 12:49 AM To: Featherstone, Annette; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Mast Cell Stain other than Toluidine Blue No, but I have two methods that are very simple, both are T blue. Churkian Schenk is my favorite, but have also used a method nicely passed on to me by Liz Chlipala out of Frieda Carson's book, a pH 4.3 T blue that takes 3 to 5 minutes, rinse with water, air dry the section and mount. This one is really simple and fast. If you want, I can forward both to you privately. At 05:37 AM 9/30/2004, you wrote: >Does anyone have a simple Mast Cell Stain other than Toluidine Blue? >Annette Featherstone HT/MLT >-----Original Message----- >From: hymclab [mailto:hymclab@hyhc.com] >Sent: Wednesday, September 29, 2004 15:23 >To: 'Andrea Grantham'; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] Bouins substitute in Trichrome staining > > >PLEASE POST > >-----Original Message----- >From: Andrea Grantham [mailto:algranth@u.arizona.edu] >Sent: Wednesday, September 29, 2004 10:38 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Bouins substitute in Trichrome staining > > >I attended a workshop at NSH last week on connective tissue staining and >one of the attendees said they were using citrate buffer instead of bouins >as a mordant. I couldn't catch up with her after the workshop to ask about >the protocol. Is anybody out in histoland doing this? Would you mind >sharing your protocol if you are? >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 > >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 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 ********************************************************************** 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 beingmary53 <@t> ev1.net Thu Sep 30 18:23:51 2004 From: beingmary53 <@t> ev1.net (maryjohnson) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Pen Message-ID: <007601c4a744$8caa4400$5a48bbd0@ugly> Hi Tracy my name is mary, if you are looking for a pen that will write on cassettes you may want to try Stat lab they have a pen that we are now using, we had the same problems. The pen ordering # is SMP-BK Stat lab # is 1-800-4423573. Wish you luck Mary From jcox90 <@t> yahoo.com Thu Sep 30 18:37:37 2004 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Pen In-Reply-To: <007601c4a744$8caa4400$5a48bbd0@ugly> Message-ID: <20040930233737.53145.qmail@web40910.mail.yahoo.com> We just tried the statlab pen in our processor and it all came off. I was told it only works on certain brands of cassettes. Test on blank cassettes before you write on regular. I was really hoping to switch but now can't. Just trying to prevent a distaster!! Jill maryjohnson wrote:Hi Tracy my name is mary, if you are looking for a pen that will write on cassettes you may want to try Stat lab they have a pen that we are now using, we had the same problems. The pen ordering # is SMP-BK Stat lab # is 1-800-4423573. Wish you luck Mary _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Jill Cox HT (ASCP) --------------------------------- Do you Yahoo!? vote.yahoo.com - Register online to vote today! From kerry.l.crabb <@t> gsk.com Thu Sep 30 19:12:40 2004 From: kerry.l.crabb <@t> gsk.com (kerry.l.crabb@gsk.com) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Histonet @ NSH S/C Message-ID: All, I appreciate your interest in the NSH Symposium/Convention and Histonet which has done so much for the histology field. I have been copied on most if not all the emails to Histonet about trying to get you all together during the S/C. Yes, that would be a great idea I believe NSH could support. Yes, it was done a number of years ago (I think twice) during Histonet's early days. Since Histonet is not an official part of NSH, a gathering for you netters is not planned or arrangements made by NSH. Although NSH does not have monies available to assist you, we may be able to assist by finding space for you to gather if someone in your group is willing to plan and make all the arrangements. The person willing to plan something for you may contact the NSH Meeting Manager (Carrie Diamond) to see if she can find a table/room for you to gather. This person may also contact me directly to discuss your ideas as they develop and the possibilities of making them happen at the next NSH S/C. Kerry Crabb NSH Convention Chairman From jhabecke <@t> seattlecca.org Thu Sep 30 19:27:22 2004 From: jhabecke <@t> seattlecca.org (Randolph-Habecker, Julie) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] More about the microbeads Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EACC0@wala01.seattlecca.org> It's me again about the microbeads. Briefly, we have some formalin-fixed lung tissue from animals who have undergone lung function tests involving the inhalation of beads - the largest being 50 microns. (We perform research histology for all of the investigators at our institution and sometimes they don't consult us before they start experiments.) Let me first thank everyone for there helpful comments about xylene alternative. This was very helpful and enlightening so we will be trying several to see how they work. Also, we are trying frozen sections. However, we would like to try to paraffin process the tissue. So.....I think I need to be more specific in my questioning. We are interested in the pathology - for that reason we do not necessarily need to see the beads in the tissue. Hear is my big question - do I want the beads to melt or not? My main concern is if the melting of the beads will alter the morphology of the tissue. My other concern is that the melted bead plastic might gum up the processor? On the other hand, if the beads are not melted but get dislodged from the tissue during processing - will that effect my processor? How will the beads effect sectioning? Is there anything else that I should consider before messing with this tissue? I would really love to talk to someone who has worked specifically with this type of tissue. Thanks, Julie Julie Randolph-Habecker, Ph.D. Experimental Histopathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave. N, G1-300 PO Box 19023 Seattle, WA 98109-1024 Tel: (206) 288-1187 FAX: (206) 288-1345 jhabecke@fhcrc.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 GREYTRUNK <@t> aol.com Thu Sep 30 20:04:04 2004 From: GREYTRUNK <@t> aol.com (GREYTRUNK@aol.com) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] florida hurricanes Message-ID: Hello everyone. I know this isn't the venue to post things like this, but it is much easier for me to write one email instead of sending several. First, I want to say that I am sorry that I was unable to go to Nationals. I know that several of you were counting on me. Sharon told me that you were concerned and asking about me at the Convention. Let me say that I am ok. Not great, but OK. I did get hit by 3 of the 4 hurricanes that went through Florida these past 6 weeks. Charley, Francis, and Jeanne all went directly over the county that I live in. Once things started to get taken care of from one hurricane, another one would come. The last one, Jeanne, came around midnight on Saturday night and my power didn't come back on until today. Hopefully it will stay on. There still are thousands of people without homes or power. As of today, the kids have missed 16 days of school already. It has been extremely stressful, but I am one of the lucky ones because I still have a home. The news reports that you may have seen do not even come close to the destruction here, not even close. A lot of businesses were destryoed as well. But organizations are helping with food, water, ice, shelter, etc. And prayers from everyone help too. Just keep praying for all of us in Polk County Florida......And that you for your concerns. I will see you next year. Roxanne Soto From katri <@t> cogeco.ca Thu Sep 30 20:18:46 2004 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] PKK1 and M2A antibodies Message-ID: <001b01c4a754$99f861e0$6a9a9618@Katri> Hi Histonetters, One of our pathologists accessed the ImmunoQuery for a panel of antibodies for a certain differential diagnoses. The panel consisted among others a PKK1 and a M2A. Among thousands of antibodies available, how am I supposed to find out who carries these particular clones? I know PKK1 is a cytokeratin antibody, but who sells it? M2A I've never heard of. I tried searches in different antibody sites (abcam and The Antibody Resource Page) with no success. Help, please! Katri Katri Tuomala St.Joseph's Healthcare Hamilton, Ontario, Canada From jkiernan <@t> uwo.ca Thu Sep 30 20:37:19 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Oxidising Agents in Haematoxylin[Scanned] References: <6AB79F0BA7C4914BA8DD8F5E0C21B97513BF37@AHEXMAIL01.xalderhey.com> Message-ID: <415CB4CF.1871C53F@uwo.ca> Yes you could use the potassium salt instead. If you oxidize with iodate, the staining solution is not Ehrlich's haematoxylin. Paul Ehrlich (1886) allowed his solution to oxidize in the air. That's why it keeps for years & years. John Kiernan London, Canada _____________________________________ Muskett David wrote: > > Dear All > > I am inquiring about the use of differing oxidising agents in histology. > I am routinely using Sodium Iodate as the oxidising agent in Ehrlich's > haematoxylin. Can the sodium iodate be substituted by Potassium Iodate? > Is there a difference in their oxidising properties? > > David > > > David Muskett > > Chief Biomedical Scientist, Histology > > Royal Liverpool Children's NHS Trust > > Eaton Road > > Liverpool > > L12 2AP > > > > Telephone (0151) 293 3656 > > Fax (0151) 293 3617 > From jkiernan <@t> uwo.ca Thu Sep 30 23:39:26 2004 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:06 2005 Subject: [Histonet] Mast Cell Stain other than Toluidine Blue References: <1CF2E2E5BB36D5119E7A0008C791F3741269E0C3@simba.kids> Message-ID: <415CDF7E.AA6FDBFE@uwo.ca> For what it's worth, I endorse Tony Henwood's comment 100%, having done mast cell research for 40+ years. Mast cells derive their identity from granules made largely of heparin, which is a heavily sulphated (=strongly acidic) macromolecular carbohydrate. At low pH, heparin and the chondroitin sulphates are just about the only big molecules (in a connective tissue) that can bind a cationic dye. Alcian blue (the genuine article) can make permanently insoluble deposits at sites of mast cell granules. Counterstains can be tried and tested ad libitum, and the turquoise alcian blue colour will always be there in the mast cell grsnules. ____________________________________________________________________ Tony Henwood wrote: > > The alcian blue at pH 1 is a good demonstration method for mast cells. > > Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager > The Children's Hospital at Westmead, > Locked Bag 4001, Westmead, 2145, AUSTRALIA. > Tel: 612 9845 3306 > Fax: 612 9845 3318 > > -----Original Message----- > From: Gayle Callis [mailto:gcallis@montana.edu] > Sent: Friday, 1 October 2004 12:49 AM > To: Featherstone, Annette; Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Mast Cell Stain other than Toluidine Blue > > No, but I have two methods that are very simple, both are T blue. Churkian > Schenk is my favorite, but have also used a method nicely passed on to me > by Liz Chlipala out of Frieda Carson's book, a pH 4.3 T blue that takes 3 > to 5 minutes, rinse with water, air dry the section and mount. This one is > really simple and fast. If you want, I can forward both to you privately. > > At 05:37 AM 9/30/2004, you wrote: > >Does anyone have a simple Mast Cell Stain other than Toluidine Blue? > >Annette Featherstone HT/MLT > >-----Original Message----- > >From: hymclab [mailto:hymclab@hyhc.com] > >Sent: Wednesday, September 29, 2004 15:23 > >To: 'Andrea Grantham'; histonet@lists.utsouthwestern.edu > >Subject: RE: [Histonet] Bouins substitute in Trichrome staining > > > > > >PLEASE POST > > > >-----Original Message----- > >From: Andrea Grantham [mailto:algranth@u.arizona.edu] > >Sent: Wednesday, September 29, 2004 10:38 AM > >To: histonet@lists.utsouthwestern.edu > >Subject: [Histonet] Bouins substitute in Trichrome staining > > > > > >I attended a workshop at NSH last week on connective tissue staining and > >one of the attendees said they were using citrate buffer instead of bouins > >as a mordant. I couldn't catch up with her after the workshop to ask about > >the protocol. Is anybody out in histoland doing this? Would you mind > >sharing your protocol if you are? > >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