From antje.marcantonio <@t> pharma.novartis.com Mon Jan 3 04:37:43 2005 From: antje.marcantonio <@t> pharma.novartis.com (antje.marcantonio@pharma.novartis.com) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] CD68 in monkey frozen tissue Message-ID: HAPPY NEW YEAR TO EVERYONE ! Hello I use the anti human CD68, KP1 from Dako on FFPE monkey tissue with good results. The same antibody is not recommended for frozens. I tried it anyway and it is indeed difficult to get an enough strong signal without background. What about the other clone PG-M1 ? Is this working on frozen sections ? Any input is very much welcome ! Antje Marcantonio Novartis Pharma AG BU Transplantation Research Basel, Switzerland Tel. +41 61 324 6730 antje.marcantonio@pharma.novartis.com From akbitting <@t> geisinger.edu Mon Jan 3 08:13:12 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] phosphorylated antibodies Message-ID: All of the phosphorylated antibodoes we are currently using for IHC on human FFPE tissue are classified as Research Use Only. Is there anyone out there selling them as IVDs? Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From STEGTM <@t> samcstl.org Mon Jan 3 11:27:51 2005 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Needle biopsies Message-ID: Ditto. I'd rather cut again, extra levels, than pare into a thready bx. On some, eg. liver, I will cut extra to begin with, anticipating specials. Peace, Terre >>> "Joe Nocito" 12/30/2004 7:00:14 AM >>> Jim, I always refer to what my 8th grade shop teacher told us. It is always easier to shave off a little more wood, than it is trying to glue it back on. I have taken this advice to the histo lab. Once, when I was a rookie many moons ago, I had to fish through paraffin shavings to locate a renal biopsy that I cut away. Since that time, I always cut on the conservative side and explain to the pathologists that it always easier to go back into the block than fish through paraffin shavings. I do get static sometimes, but after they cool down, they realize it was a good descion. As always, the opinions of the author do not reflect the opinions of his employers and their lawyers. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jim Ball Sent: Wednesday, December 29, 2004 3:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Needle biopsies I am a tech with 25+ years of experience and have been bitten by about every snake in the garden of eden (Histology), and I guess that is one of the main reasons I will error on the side of caution at every turn. I really try to be as conservative as possible with tissue when trimming into a needle biopsy, as soon as I have a full face on properly enbedded needles(usually not more than 20 microns or less I start taking slides). The sections are 3microns and may produce as many as 5 to 10 sections suitable for mounting. This acounts for max 30 more micrones into the block. It is at this point I would like to preserve the remainder of the tissue until it is reviewed by a pathologist. I refer to my madness as scouting (a procedure if used by General Custer would have saved alot of lives), but as we all know there are some patologist that will declare we did not trim enough if what they are looking for is 100 micrones into the block. While I have been reseaching a procedure that will keep everyone happy I ran across an article that state there was a study done to determine if histologists were trimming away microcalcifications in needle biopsies, and according to the high lights of the article (one they wanted me to purchase to add insult to injury) it was determined that after x-raying the histology shavings from trimmed breast biopsies the culprit once again was the histologist. Go figure. At the present time I am on a public computer and some one needs to use it, but before I leave please foward any ideas you may have on this subject via this server or directly to my e-mail address listed with this posting _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From japoteete <@t> saintfrancis.com Mon Jan 3 11:39:30 2005 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Needle biopsies Message-ID: Cutting thru a block is like asking for an automatic lawsuit these days. The pathologists may fuss on occasion if they want deeper sections and must wait for them, but if it were my tissue, I would prefer that the Histologists be conservative. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Saint Francis Hospital Tulsa OK japoteete@saintfrancis.com -----Original Message----- From: Therersa Stegall [mailto:STEGTM@samcstl.org] Sent: Monday, January 03, 2005 11:28 AM To: histo007@hotmail.com; histonet@lists.utsouthwestern.edu; JNocito@Pathreflab.com Subject: RE: [Histonet] Needle biopsies Ditto. I'd rather cut again, extra levels, than pare into a thready bx. On some, eg. liver, I will cut extra to begin with, anticipating specials. Peace, Terre >>> "Joe Nocito" 12/30/2004 7:00:14 AM >>> Jim, I always refer to what my 8th grade shop teacher told us. It is always easier to shave off a little more wood, than it is trying to glue it back on. I have taken this advice to the histo lab. Once, when I was a rookie many moons ago, I had to fish through paraffin shavings to locate a renal biopsy that I cut away. Since that time, I always cut on the conservative side and explain to the pathologists that it always easier to go back into the block than fish through paraffin shavings. I do get static sometimes, but after they cool down, they realize it was a good descion. As always, the opinions of the author do not reflect the opinions of his employers and their lawyers. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jim Ball Sent: Wednesday, December 29, 2004 3:56 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Needle biopsies I am a tech with 25+ years of experience and have been bitten by about every snake in the garden of eden (Histology), and I guess that is one of the main reasons I will error on the side of caution at every turn. I really try to be as conservative as possible with tissue when trimming into a needle biopsy, as soon as I have a full face on properly enbedded needles(usually not more than 20 microns or less I start taking slides). The sections are 3microns and may produce as many as 5 to 10 sections suitable for mounting. This acounts for max 30 more micrones into the block. It is at this point I would like to preserve the remainder of the tissue until it is reviewed by a pathologist. I refer to my madness as scouting (a procedure if used by General Custer would have saved alot of lives), but as we all know there are some patologist that will declare we did not trim enough if what they are looking for is 100 micrones into the block. While I have been reseaching a procedure that will keep everyone happy I ran across an article that state there was a study done to determine if histologists were trimming away microcalcifications in needle biopsies, and according to the high lights of the article (one they wanted me to purchase to add insult to injury) it was determined that after x-raying the histology shavings from trimmed breast biopsies the culprit once again was the histologist. Go figure. At the present time I am on a public computer and some one needs to use it, but before I leave please foward any ideas you may have on this subject via this server or directly to my e-mail address listed with this posting _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ 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 sluhisto <@t> yahoo.com Mon Jan 3 12:27:29 2005 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] fluorescent filter slider housing Message-ID: <20050103182729.71846.qmail@web51008.mail.yahoo.com> I am looking for a flourescent filter housing unit for my Zeis standard research microscope (vintage 1980). I have one slider with installed flourescein and rhodamine filter sets but want a second slider to permanently hold a DAPI filter set (rather than having to unscrew one of the filter sets in my present housing unit and install the DAPI whenever I want to use DAPI). The piece I'm looking for is the piece that slides into the cylindrical tube port just above and to the right of the objective lenses on these scopes equipped for fluorescence microscopy. The number stamped on the unit is 46 63 01-9901. They want $600 for a new one so I'm hoping to find someone who has one of these things which they no longer use and would be willing to sell (or give) to me. Please get in touch if you have such a thing and thanks in advance. Jan Ryerse ryersejs@slu.edu 314-977-7848 St. Louis University Health Sciences Center Dept of Pathology __________________________________ Do you Yahoo!? Yahoo! Mail - now with 250MB free storage. Learn more. http://info.mail.yahoo.com/mail_250 From pruegg <@t> ihctech.net Mon Jan 3 13:30:56 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] two of every message Message-ID: <200501031930.j03JUbAs027089@chip.viawest.net> Hey guys how do I tell someone that I am receiving two of every message now that I just resubscribed. Patsy Ruegg pruegg@ihctech.net From cmmathis <@t> wfubmc.edu Mon Jan 3 15:51:11 2005 From: cmmathis <@t> wfubmc.edu (Cathy M. Mathis) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Frozen section of plastic stent Message-ID: <9AEEF1FB6254224AA355ED285F84916508454C5B@EXCHVS2.medctr.ad.wfubmc.edu> Happy New Year Histonetters!! Any suggestions for cutting plastic (silicone) stent embedded in OCT on the cryostat. I'm cutting at 10 microns, -28 C and about every third or fourth section is good. I am moving the blade every 20-30 sections. Most of the sections are badly folded or fractured. The stent will eventually be part of tissue, farther into the block. Thanks in advance for the help. Cathy Cathy M. Mathis, H.T. (ASCP) Institute of Regenerative Medicine Wake Forest University Health Sciences Tissue Engineering Laboratory NRC Bldg. Rm. 126 Medical Center Blvd. Winston-Salem, NC 27157 ph: 336-713-7285 email: cmmathis@wfubmc.edu From ftulenko06 <@t> jcu.edu Mon Jan 3 15:57:32 2005 From: ftulenko06 <@t> jcu.edu (ftulenko06@jcu.edu) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] (no subject) Message-ID: Hello everybody, I was wondering how long paraplast sections that have been mounted on slides could be left alone after drying before staining. Is there any reason they couldn't be left for days or weeks? Thanks for your advice, Frank From pruegg <@t> ihctech.net Mon Jan 3 16:11:51 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Frozen section of plastic stent In-Reply-To: <9AEEF1FB6254224AA355ED285F84916508454C5B@EXCHVS2.medctr.ad.wfubmc.edu> Message-ID: <200501032211.j03MBWAs023167@chip.viawest.net> Cathy I would use the tape transfer system from Instrumedics to hold everything together but you will probably need to use steel or tungsten carbide knives. Patsy Ruegg -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cathy M. Mathis Sent: Monday, January 03, 2005 2:51 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Frozen section of plastic stent Happy New Year Histonetters!! Any suggestions for cutting plastic (silicone) stent embedded in OCT on the cryostat. I'm cutting at 10 microns, -28 C and about every third or fourth section is good. I am moving the blade every 20-30 sections. Most of the sections are badly folded or fractured. The stent will eventually be part of tissue, farther into the block. Thanks in advance for the help. Cathy Cathy M. Mathis, H.T. (ASCP) Institute of Regenerative Medicine Wake Forest University Health Sciences Tissue Engineering Laboratory NRC Bldg. Rm. 126 Medical Center Blvd. Winston-Salem, NC 27157 ph: 336-713-7285 email: cmmathis@wfubmc.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eileen_dusek <@t> yahoo.com Mon Jan 3 16:48:06 2005 From: eileen_dusek <@t> yahoo.com (eileen dusek) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Ideas for the Illinois Society Message-ID: <20050103224806.67306.qmail@web11903.mail.yahoo.com> Hi everyone, I hope you all had a great and wonderful Holiday with family and friends. I am starting to plan for the 2006 Illinois state meeting. We have continued to have wonderful speakers through out the years. I am trying to come up with new topics. If you were to go your state meeting what discussion or workshop would you be drawn to? Some thought I have are Basic Staining, Cytology/Histology correlations, FNA's, Plant Pathology, and Veterinary Pathology. I would love to hear from you, perhaps this will help other states also. Thank you Eileen C. Dusek Edward Hospital Naperville, Ill. --------------------------------- Do you Yahoo!? Dress up your holiday email, Hollywood style. Learn more. From cforster <@t> umn.edu Mon Jan 3 17:10:01 2005 From: cforster <@t> umn.edu (Colleen Forster) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] long term storage of specimens Message-ID: <41D9D0C9.9020300@umn.edu> Histonetters, I have been asked to post a question: A museum has tissue samples for teaching stored in 10% formalin. They are wondering what other solution they might be able to use that would be less toxic. This would take the place of the formalin for long term storage. I believe they take the samples out for demonstration so putting them in a plastic is not an option. Any suggestions???? Colleen Forster U of Mn 612-626-0436 -- Outgoing mail is certified Virus Free. Checked by AVG Anti-Virus (http://www.grisoft.com). Version: 7.0.279 / Virus Database: 265.6.7 - Release Date: 12/30/2004 From Janette_Thurley <@t> health.qld.gov.au Mon Jan 3 18:01:32 2005 From: Janette_Thurley <@t> health.qld.gov.au (Janette Thurley) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] long term storage of specimens Message-ID: Have you considered plastination? The specimen is permanently inpregnated with a plastic. This removes the need for storing in any solution. Try contacting Anatomical Pathology at the University of Queensland for more details. It has been done by Robbie Boyse on whole organs quite successfully. Janette Thurley Chief Scientist Anatomical Pathology and Cytopathology Queensland Health Pathology Service janette_thurley@health.qld.gov.au >>> Colleen Forster 4/01/05 9:10:01 >>> Histonetters, I have been asked to post a question: A museum has tissue samples for teaching stored in 10% formalin. They are wondering what other solution they might be able to use that would be less toxic. This would take the place of the formalin for long term storage. I believe they take the samples out for demonstration so putting them in a plastic is not an option. Any suggestions???? Colleen Forster U of Mn 612-626-0436 -- Outgoing mail is certified Virus Free. Checked by AVG Anti-Virus (http://www.grisoft.com). Version: 7.0.279 / Virus Database: 265.6.7 - Release Date: 12/30/2004 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet *********************************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is prohibited. It may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone or by return email. You should also delete this email and destroy any hard copies produced. *********************************************************************************** From c.m.vanderloos <@t> amc.uva.nl Tue Jan 4 01:37:58 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] RE: CD68 in monkey frozen tissue Message-ID: <700d19703128.703128700d19@amc.uva.nl> A HAPPY AND HEALTHY NEW YEAR TO EVERYONE TOO! CD68, clone PG-M1 does stain something on frozen sections. However, if this staining result reflects the full spectrum of macrophages is a bit doubtful. For frozens you better use CD68 clone EBM11. Best wishes, Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: c.m.vanderloos@amc.uva.nl ----- Original Message ----- >From antje.marcantonio@pharma.novartis.com Date Mon, 3 Jan 2005 11:37:43 +0100 To histonet@lists.utsouthwestern.edu Subject [Histonet] CD68 in monkey frozen tissue HAPPY NEW YEAR TO EVERYONE ! Hello I use the anti human CD68, KP1 from Dako on FFPE monkey tissue with good results. The same antibody is not recommended for frozens. I tried it anyway and it is indeed difficult to get an enough strong signal without background. What about the other clone PG-M1 ? Is this working on frozen sections ? Any input is very much welcome ! Antje Marcantonio Novartis Pharma AG BU Transplantation Research Basel, Switzerland Tel. +41 61 324 6730 antje.marcantonio@pharma.novartis.com From abright <@t> brightinstruments.com Tue Jan 4 04:47:20 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Frozen section of plastic stent Message-ID: Dear Cathy & others with this problem, I do not know the answer to your problem with sectioning plastic (silicone) stents, but I have experience of sectioning a wide range of materials including plastics. If you could send me a sample I would be very happy to section the stent and give you a full report with no costs to you. I am hoping that all that is required is determining the correct temperature and maybe using a tungsten carbide tipped knife. 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: Cathy M. Mathis [mailto:cmmathis@wfubmc.edu] Sent: 03 January 2005 21:51 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Frozen section of plastic stent Happy New Year Histonetters!! Any suggestions for cutting plastic (silicone) stent embedded in OCT on the cryostat. I'm cutting at 10 microns, -28 C and about every third or fourth section is good. I am moving the blade every 20-30 sections. Most of the sections are badly folded or fractured. The stent will eventually be part of tissue, farther into the block. Thanks in advance for the help. Cathy Cathy M. Mathis, H.T. (ASCP) Institute of Regenerative Medicine Wake Forest University Health Sciences Tissue Engineering Laboratory NRC Bldg. Rm. 126 Medical Center Blvd. Winston-Salem, NC 27157 ph: 336-713-7285 email: cmmathis@wfubmc.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Tue Jan 4 06:14:14 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] looking for slide dryer[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EDE2@bhrv-nt-11.bhrv.nwest.nhs.uk> Do you still have all your own hair Terry? Or are being macho and not using a hair drier cos you're bald? I personally use one daily as I've still got my hair despite it being a funny colour. Don't trash Nancy Boys, it's not nice. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 29 December 2004 16:28 To: Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] I have often wondered if anyone has rigged a perforated box to one of those elephant trunk women's hair driers to produce a slide dryer. Come to think of it, men probably use them to, they become such a bunch of Nancy boys. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kapoor, Sue [mailto:Sue.Kapoor@uhsi.org] Sent: 29 December 2004 16:03 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] looking for slide dryer Hi all, I'm looking to purchase a slide dryer, I would prefer the type that you open and set a rack of slides into, not the type that you lay the slides on. Demo units, rebuilts will be fine, the only catch is it MUST be under $500, can anyone help me? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From wayneholland1959 <@t> msn.com Tue Jan 4 09:00:25 2005 From: wayneholland1959 <@t> msn.com (Wayne Holland) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Sample questions for HT and QIHC Message-ID: Can anyone give me information on how to get sample questions for the HT and QIHC examinations? Thanks so much! _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfeeŽ Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From cfavara <@t> niaid.nih.gov Tue Jan 4 09:14:46 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] long term storage of specimens Message-ID: Janette, A million years ago before the era of litigation we used to take whole organs and impregnate with paraffin. These were usually hearts with malformations that had been sectioned to show the malformation. If you are interested I can try to find more information. Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Janette Thurley [mailto:Janette_Thurley@health.qld.gov.au] Sent: Monday, January 03, 2005 5:02 PM To: histonet@lists.utsouthwestern.edu; cforster@umn.edu Subject: Re: [Histonet] long term storage of specimens Have you considered plastination? The specimen is permanently inpregnated with a plastic. This removes the need for storing in any solution. Try contacting Anatomical Pathology at the University of Queensland for more details. It has been done by Robbie Boyse on whole organs quite successfully. Janette Thurley Chief Scientist Anatomical Pathology and Cytopathology Queensland Health Pathology Service janette_thurley@health.qld.gov.au >>> Colleen Forster 4/01/05 9:10:01 >>> Histonetters, I have been asked to post a question: A museum has tissue samples for teaching stored in 10% formalin. They are wondering what other solution they might be able to use that would be less toxic. This would take the place of the formalin for long term storage. I believe they take the samples out for demonstration so putting them in a plastic is not an option. Any suggestions???? Colleen Forster U of Mn 612-626-0436 -- Outgoing mail is certified Virus Free. Checked by AVG Anti-Virus (http://www.grisoft.com). Version: 7.0.279 / Virus Database: 265.6.7 - Release Date: 12/30/2004 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet **************************************************************************** ******* This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is prohibited. It may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone or by return email. You should also delete this email and destroy any hard copies produced. **************************************************************************** ******* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sue.Kapoor <@t> uhsi.org Tue Jan 4 09:17:05 2005 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] large cryostat chucks Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F5693@khmcexch.uhsi.org> Hi everyone, I'm looking for large 1 1/2 inch round crystat chucks for my Leica cryostat, can anyone suggest vendors? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From japoteete <@t> saintfrancis.com Tue Jan 4 09:18:29 2005 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Sample questions for HT and QIHC Message-ID: My QIHC exam in the fall of 2003 was all "practical projects" and had only one question. The answer was to be submitted with the slides. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Lab Saint Francis Hospital Tulsa, OK japoteete@saintfrancis.com -----Original Message----- From: Wayne Holland [mailto:wayneholland1959@msn.com] Sent: Tuesday, January 04, 2005 9:00 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Sample questions for HT and QIHC Can anyone give me information on how to get sample questions for the HT and QIHC examinations? Thanks so much! _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee(r) Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 _______________________________________________ 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 mari.ann.mailhiot <@t> leica-microsystems.com Tue Jan 4 09:32:57 2005 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] large cryostat chucks Message-ID: Hi Sue I beleive our 40 mm specimen disc is what you need. The part number is 14037008637. You can order the disc directly though Leica at 800 248 0123 option 1. If you need further assistance just give me a call. Best Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com "Kapoor, Sue" To: histonet@lists.utsouthwestern.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] large cryostat chucks western.edu 01/04/2005 09:17 AM Hi everyone, I'm looking for large 1 1/2 inch round crystat chucks for my Leica cryostat, can anyone suggest vendors? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From Terry.Marshall <@t> rothgen.nhs.uk Tue Jan 4 09:53:06 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] looking for slide dryer[Scanned] Message-ID: Kemlo wrote: "Do you still have all your own hair Terry? Or are being macho and not using a hair drier cos you're bald?" Yes, all my hair is my own. "I personally use one daily as I've still got my hair despite it being a funny colour." A/ Hmm. I'd love to see a picture of you sitting there with this elephant trunk on your head. Do you read Women's Own? B/ Purple? "Don't trash Nancy Boys, it's not nice." Well, you would be quick to tell me wouldn't you, but in fact, I didn't do any trashing, merely made a passing observation that men had become what I regard as effeminate. Earrings necklaces, perfumes and other cosmetics, sales of which approximated to female cosmetics in 2003. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 04 January 2005 12:14 To: Marshall Terry Dr, Consultant Histopathologist; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 29 December 2004 16:28 To: Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] I have often wondered if anyone has rigged a perforated box to one of those elephant trunk women's hair driers to produce a slide dryer. Come to think of it, men probably use them to, they become such a bunch of Nancy boys. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kapoor, Sue [mailto:Sue.Kapoor@uhsi.org] Sent: 29 December 2004 16:03 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] looking for slide dryer Hi all, I'm looking to purchase a slide dryer, I would prefer the type that you open and set a rack of slides into, not the type that you lay the slides on. Demo units, rebuilts will be fine, the only catch is it MUST be under $500, can anyone help me? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Tue Jan 4 10:00:55 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] looking for slide dryer[Scanned] Message-ID: <83AACDB0810528418AA106F9AE9B7F7E50780F@sjhaexc02.sjha.org> ok - What are Nancy boys??? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Tuesday, January 04, 2005 10:53 AM To: Kemlo Rogerson; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] Kemlo wrote: "Do you still have all your own hair Terry? Or are being macho and not using a hair drier cos you're bald?" Yes, all my hair is my own. "I personally use one daily as I've still got my hair despite it being a funny colour." A/ Hmm. I'd love to see a picture of you sitting there with this elephant trunk on your head. Do you read Women's Own? B/ Purple? "Don't trash Nancy Boys, it's not nice." Well, you would be quick to tell me wouldn't you, but in fact, I didn't do any trashing, merely made a passing observation that men had become what I regard as effeminate. Earrings necklaces, perfumes and other cosmetics, sales of which approximated to female cosmetics in 2003. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 04 January 2005 12:14 To: Marshall Terry Dr, Consultant Histopathologist; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 29 December 2004 16:28 To: Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] I have often wondered if anyone has rigged a perforated box to one of those elephant trunk women's hair driers to produce a slide dryer. Come to think of it, men probably use them to, they become such a bunch of Nancy boys. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kapoor, Sue [mailto:Sue.Kapoor@uhsi.org] Sent: 29 December 2004 16:03 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] looking for slide dryer Hi all, I'm looking to purchase a slide dryer, I would prefer the type that you open and set a rack of slides into, not the type that you lay the slides on. Demo units, rebuilts will be fine, the only catch is it MUST be under $500, can anyone help me? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@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 Janet.Bonner <@t> FLHOSP.ORG Tue Jan 4 10:02:08 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] large cryostat chucks Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4144@fh2k093.fhmis.net> Try the company "MicroOptics" . Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 1/4/2005 10:17 AM Subject: [Histonet] large cryostat chucks Hi everyone, I'm looking for large 1 1/2 inch round crystat chucks for my Leica cryostat, can anyone suggest vendors? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 ajennings <@t> unmc.edu Tue Jan 4 10:04:05 2005 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] looking for slide dryer[Scanned] back on topic please In-Reply-To: Message-ID: Please keep this professional guys and back on topic, or take it to private emails Thank you Anita "Marshall Terry Dr, Consultant To Histopathologist" "Kemlo Rogerson" , othgen.nhs.uk> "Kapoor, Sue" Sent by: , histonet-bounces@ lists.utsouthwest cc ern.edu Subject RE: [Histonet] looking for slide 01/04/2005 09:53 dryer[Scanned] AM Kemlo wrote: "Do you still have all your own hair Terry? Or are being macho and not using a hair drier cos you're bald?" Yes, all my hair is my own. "I personally use one daily as I've still got my hair despite it being a funny colour." A/ Hmm. I'd love to see a picture of you sitting there with this elephant trunk on your head. Do you read Women's Own? B/ Purple? "Don't trash Nancy Boys, it's not nice." Well, you would be quick to tell me wouldn't you, but in fact, I didn't do any trashing, merely made a passing observation that men had become what I regard as effeminate. Earrings necklaces, perfumes and other cosmetics, sales of which approximated to female cosmetics in 2003. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 04 January 2005 12:14 To: Marshall Terry Dr, Consultant Histopathologist; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 29 December 2004 16:28 To: Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] I have often wondered if anyone has rigged a perforated box to one of those elephant trunk women's hair driers to produce a slide dryer. Come to think of it, men probably use them to, they become such a bunch of Nancy boys. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kapoor, Sue [mailto:Sue.Kapoor@uhsi.org] Sent: 29 December 2004 16:03 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] looking for slide dryer Hi all, I'm looking to purchase a slide dryer, I would prefer the type that you open and set a rack of slides into, not the type that you lay the slides on. Demo units, rebuilts will be fine, the only catch is it MUST be under $500, can anyone help me? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From japoteete <@t> saintfrancis.com Tue Jan 4 10:18:06 2005 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] looking for slide dryer[Scanned] Message-ID: Oh, for the good old days, when the prime semi-off-the-subject topics involved the best Scotch and pubs, and we knew what all the words meant. Jacquie Poteete MT(ASCP)QIHC Tulsa, OK -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Tuesday, January 04, 2005 10:01 AM To: Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] ok - What are Nancy boys??? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Tuesday, January 04, 2005 10:53 AM To: Kemlo Rogerson; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] Kemlo wrote: "Do you still have all your own hair Terry? Or are being macho and not using a hair drier cos you're bald?" Yes, all my hair is my own. "I personally use one daily as I've still got my hair despite it being a funny colour." A/ Hmm. I'd love to see a picture of you sitting there with this elephant trunk on your head. Do you read Women's Own? B/ Purple? "Don't trash Nancy Boys, it's not nice." Well, you would be quick to tell me wouldn't you, but in fact, I didn't do any trashing, merely made a passing observation that men had become what I regard as effeminate. Earrings necklaces, perfumes and other cosmetics, sales of which approximated to female cosmetics in 2003. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 04 January 2005 12:14 To: Marshall Terry Dr, Consultant Histopathologist; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 29 December 2004 16:28 To: Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] I have often wondered if anyone has rigged a perforated box to one of those elephant trunk women's hair driers to produce a slide dryer. Come to think of it, men probably use them to, they become such a bunch of Nancy boys. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kapoor, Sue [mailto:Sue.Kapoor@uhsi.org] Sent: 29 December 2004 16:03 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] looking for slide dryer Hi all, I'm looking to purchase a slide dryer, I would prefer the type that you open and set a rack of slides into, not the type that you lay the slides on. Demo units, rebuilts will be fine, the only catch is it MUST be under $500, can anyone help me? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@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. ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From Terry.Marshall <@t> rothgen.nhs.uk Tue Jan 4 10:20:47 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] looking for slide dryer[Scanned] Message-ID: Funny how some words/expressions are semi-country specific, and of course, not always obviously so. I had to look up (from another list) what a "hickey" was less than a week ago. 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: Poteete, Jacquie A. [mailto:japoteete@saintfrancis.com] Sent: 04 January 2005 16:18 To: 'Weems, Joyce'; Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] Oh, for the good old days, when the prime semi-off-the-subject topics involved the best Scotch and pubs, and we knew what all the words meant. From Kemlo.Rogerson <@t> elht.nhs.uk Tue Jan 4 10:30:16 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] looking for slide dryer[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EDE4@bhrv-nt-11.bhrv.nwest.nhs.uk> Nancy boys? What do you call them over there? Fags, that's it! Over here Fags are either put in your mouth and smoked or they wash your smalls. Um.......... Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Poteete, Jacquie A. [mailto:japoteete@saintfrancis.com] Sent: 04 January 2005 16:18 To: 'Weems, Joyce'; Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] Oh, for the good old days, when the prime semi-off-the-subject topics involved the best Scotch and pubs, and we knew what all the words meant. Jacquie Poteete MT(ASCP)QIHC Tulsa, OK -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Tuesday, January 04, 2005 10:01 AM To: Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] ok - What are Nancy boys??? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Tuesday, January 04, 2005 10:53 AM To: Kemlo Rogerson; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] Kemlo wrote: "Do you still have all your own hair Terry? Or are being macho and not using a hair drier cos you're bald?" Yes, all my hair is my own. "I personally use one daily as I've still got my hair despite it being a funny colour." A/ Hmm. I'd love to see a picture of you sitting there with this elephant trunk on your head. Do you read Women's Own? B/ Purple? "Don't trash Nancy Boys, it's not nice." Well, you would be quick to tell me wouldn't you, but in fact, I didn't do any trashing, merely made a passing observation that men had become what I regard as effeminate. Earrings necklaces, perfumes and other cosmetics, sales of which approximated to female cosmetics in 2003. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 04 January 2005 12:14 To: Marshall Terry Dr, Consultant Histopathologist; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 29 December 2004 16:28 To: Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] I have often wondered if anyone has rigged a perforated box to one of those elephant trunk women's hair driers to produce a slide dryer. Come to think of it, men probably use them to, they become such a bunch of Nancy boys. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kapoor, Sue [mailto:Sue.Kapoor@uhsi.org] Sent: 29 December 2004 16:03 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] looking for slide dryer Hi all, I'm looking to purchase a slide dryer, I would prefer the type that you open and set a rack of slides into, not the type that you lay the slides on. Demo units, rebuilts will be fine, the only catch is it MUST be under $500, can anyone help me? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@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. ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From vazquezr <@t> ohsu.edu Tue Jan 4 10:42:39 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] large cryostat chucks Message-ID: Sue, I get them from Thermo Electron Corp (formly Shandon) 1-800-547-7429, thet maybe able to help you. Robyn Surgical Derm Oregon Health & Science University >>> "Kapoor, Sue" 1/4/2005 7:17:05 AM >>> Hi everyone, I'm looking for large 1 1/2 inch round crystat chucks for my Leica cryostat, can anyone suggest vendors? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Tue Jan 4 10:46:30 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] looking for slide dryer[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EDE5@bhrv-nt-11.bhrv.nwest.nhs.uk> Are you still having problems coping with your feminine side Terry? Just because you use deodorants and a hairdryer doesn't mean you bat for the other side; I worry about you sometimes. Do you still use Hai Karate and Old Spice? Personally I blame the water for the reduced sperm count in younger males and the increasing limpness of their wrists; especially London water that's gone through countless kidneys. heheheheeh Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 04 January 2005 15:53 To: Kemlo Rogerson; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] Kemlo wrote: "Do you still have all your own hair Terry? Or are being macho and not using a hair drier cos you're bald?" Yes, all my hair is my own. "I personally use one daily as I've still got my hair despite it being a funny colour." A/ Hmm. I'd love to see a picture of you sitting there with this elephant trunk on your head. Do you read Women's Own? B/ Purple? "Don't trash Nancy Boys, it's not nice." Well, you would be quick to tell me wouldn't you, but in fact, I didn't do any trashing, merely made a passing observation that men had become what I regard as effeminate. Earrings necklaces, perfumes and other cosmetics, sales of which approximated to female cosmetics in 2003. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 04 January 2005 12:14 To: Marshall Terry Dr, Consultant Histopathologist; Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 29 December 2004 16:28 To: Kapoor, Sue; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] looking for slide dryer[Scanned] I have often wondered if anyone has rigged a perforated box to one of those elephant trunk women's hair driers to produce a slide dryer. Come to think of it, men probably use them to, they become such a bunch of Nancy boys. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kapoor, Sue [mailto:Sue.Kapoor@uhsi.org] Sent: 29 December 2004 16:03 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] looking for slide dryer Hi all, I'm looking to purchase a slide dryer, I would prefer the type that you open and set a rack of slides into, not the type that you lay the slides on. Demo units, rebuilts will be fine, the only catch is it MUST be under $500, can anyone help me? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Tue Jan 4 10:49:18 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] large cryostat chucks Message-ID: Sue, Also, if they are too long on the post, I requistioned for a maintenence person to cut them down to the correct length or yourself at home. Just a hacksaw and a bench vise is all that is needed. Robyn OHSU >>> "Kapoor, Sue" 1/4/2005 7:17:05 AM >>> Hi everyone, I'm looking for large 1 1/2 inch round crystat chucks for my Leica cryostat, can anyone suggest vendors? thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GoodwinD <@t> pahosp.com Tue Jan 4 10:55:25 2005 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Block and slide labelers--again! Message-ID: <992899E9EC268548AB8DDE246AF88473055F4CC9@PAHEX01.uphs.upenn.edu> Happy New Year, Histonet! Particulary in the NY, NJ, PA and DE areas (but comments are welcome from everywhere, of course!), which cassette and slide labelers does everyone like, and which ones are a must to avoid and why? Thanks in advance, Diana G. Goodwin Department of Pathology Pennsylvania Hospital 800 Spruce St., Preston 655-C Philadelphia, PA 19107 ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind2pahosp.com From kbroomal <@t> NEMOURS.ORG Tue Jan 4 12:04:09 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Sample questions for HT and QIHC Message-ID: <9BCAC308B27CAD4196D8AC9ADF037AAA110A78EF@wlmmsx01.nemours.org> Practice Questions for the Histotechnology Examinations: Board of Registry Study Guide by Freida L. Carson http://www.amazon.com/exec/obidos/ASIN/089189473X/qid=1104861433/sr=2-3/ref= pd_ka_b_2_3/002-0984525-6456835 Good Luck Kristen -----Original Message----- From: Wayne Holland [mailto:wayneholland1959@msn.com] Sent: Tuesday, January 04, 2005 10:00 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Sample questions for HT and QIHC Can anyone give me information on how to get sample questions for the HT and QIHC examinations? Thanks so much! _________________________________________________________________ Is your PC infected? Get a FREE online computer virus scan from McAfee? Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From phesch <@t> charter.net Tue Jan 4 12:17:30 2005 From: phesch <@t> charter.net (Pam Hesch) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Block and slide labelers--again! References: <992899E9EC268548AB8DDE246AF88473055F4CC9@PAHEX01.uphs.upenn.edu> Message-ID: <003f01c4f289$a7e39dd0$6097b418@yourae066c3a9b> Diana, I have worked extensively with the cassette printers from Leica, we had 6 where I worked and there was very few problems. They are very fast! Ours are interfaces with Co-Path+ Pam ----- Original Message ----- From: "Goodwin, Diana" To: "Histonet (E-mail)" Sent: Tuesday, January 04, 2005 10:55 AM Subject: [Histonet] Block and slide labelers--again! > Happy New Year, Histonet! > > Particulary in the NY, NJ, PA and DE areas (but comments are welcome from > everywhere, of course!), which cassette and slide labelers does everyone > like, and which ones are a must to avoid and why? > > Thanks in advance, > Diana G. Goodwin > Department of Pathology > Pennsylvania Hospital > 800 Spruce St., Preston 655-C > Philadelphia, PA 19107 > > ph: 215-829-6532 > fax: 215-829-7564 > e-mail: goodwind2pahosp.com > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Wanda.Smith <@t> HCAhealthcare.com Tue Jan 4 14:40:03 2005 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] The "Stuff" to Find Lymph Nodes Message-ID: <8784A3EE34199644AF60DBE517F5B0A60A8A571C@louex04.lou.medcity.net> Dear Histonetters, My mind has gone blank...and I need help please! What are some of the commercial solutions available to help ID lymph nodes at the grossing table? What are your experiences, pro and con, of these solutions??? One of my Pathologist has read a few articles concerning finding lymph nodes on colon cancer cases. A high number of lymph nodes must be found, down to the 1-2 mm size nodes and we are interested in other's experiences. Thanks for any and all info!!! 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 liz <@t> premierlab.com Tue Jan 4 16:38:45 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] contract TEM lab Message-ID: <000001c4f2ae$2659d7f0$76d48a80@AMY> I remember a recent post about a new contract EM lab. I have a client that might need some TEM work done is there anyone out there that does this work for research organizations. Could you e-mail me back with contact information. Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From wolf <@t> webhart.net Tue Jan 4 17:58:48 2005 From: wolf <@t> webhart.net (Patrick Paulusse) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] The "Stuff" to Find Lymph Nodes References: <8784A3EE34199644AF60DBE517F5B0A60A8A571C@louex04.lou.medcity.net> Message-ID: <000901c4f2b9$564a54c0$63b05bd1@wolf> Wanda, Although I have not been fortunate enough to have tried some of the commercially available lymph node retrieval solutions, I do use a homemade solution which was outlined in an article (referenced below) that you might be interested in trying. The recipe is as follows: 500 ml of absolute ethanol 170 ml distilled water 80 ml of 37% formaldehyde 50 ml of acetic acid According to the article the reagents must be added in this order (does anyone know why?). I've used this since 2001 with great success. I dissect off the pericolic adipose tissue and leave it sit in the solution for 24 hours and the smallest of lymph nodes show up white against the grey/yellow adipose tissue. I've also let it sit over the weekend and have seen no deleterious affects because of the prolonged treatment. If you try it let me know what you think. Article Reference: "GEWF solution: an inexpensive, simple, and effectiveaid for the retrieval of lymph nodes from colorectal cancer specimens". Ken J. Newell, Barry W. Sawka, Brian F. Rudick, David K. Driman. Arch Pathol Lab Medicine, May 2001;125:642-645. Patrick Paulusse Histology Supervisor Pembroke Regional Hospital Ontario, Canada ----- Original Message ----- From: "Smith Wanda" To: Sent: Tuesday, January 04, 2005 3:40 PM Subject: [Histonet] The "Stuff" to Find Lymph Nodes > Dear Histonetters, > My mind has gone blank...and I need help please! > What are some of the commercial solutions available to help ID lymph nodes > at the grossing table? What are your experiences, pro and con, of these > solutions??? > One of my Pathologist has read a few articles concerning finding lymph nodes > on colon cancer cases. A high number of lymph nodes must be found, down to > the 1-2 mm size nodes and we are interested in other's experiences. > Thanks for any and all info!!! > 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 cfavara <@t> niaid.nih.gov Tue Jan 4 18:12:31 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] The "Stuff" to Find Lymph Nodes Message-ID: Wanda, This is similar to Davidson Fixative that we use here for eyes. I think the order is important for two reasons but would not want to be tested on either. Impurities are added to absolute ETOH and depending on what is added for this purpose a precipitate may form when it is added to acidic solutions. Formaldehyde will do the same. This is based on my very old knowledge and many years of experience. In my experience the precipitate forms in the solution interface and does not always redissolve. I am confident someone will have a more scientific explanation and I look forward to reading. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Patrick Paulusse [mailto:wolf@webhart.net] Sent: Tuesday, January 04, 2005 4:59 PM To: Histonet; Smith Wanda Subject: Re: [Histonet] The "Stuff" to Find Lymph Nodes Wanda, Although I have not been fortunate enough to have tried some of the commercially available lymph node retrieval solutions, I do use a homemade solution which was outlined in an article (referenced below) that you might be interested in trying. The recipe is as follows: 500 ml of absolute ethanol 170 ml distilled water 80 ml of 37% formaldehyde 50 ml of acetic acid According to the article the reagents must be added in this order (does anyone know why?). I've used this since 2001 with great success. I dissect off the pericolic adipose tissue and leave it sit in the solution for 24 hours and the smallest of lymph nodes show up white against the grey/yellow adipose tissue. I've also let it sit over the weekend and have seen no deleterious affects because of the prolonged treatment. If you try it let me know what you think. Article Reference: "GEWF solution: an inexpensive, simple, and effectiveaid for the retrieval of lymph nodes from colorectal cancer specimens". Ken J. Newell, Barry W. Sawka, Brian F. Rudick, David K. Driman. Arch Pathol Lab Medicine, May 2001;125:642-645. Patrick Paulusse Histology Supervisor Pembroke Regional Hospital Ontario, Canada ----- Original Message ----- From: "Smith Wanda" To: Sent: Tuesday, January 04, 2005 3:40 PM Subject: [Histonet] The "Stuff" to Find Lymph Nodes > Dear Histonetters, > My mind has gone blank...and I need help please! > What are some of the commercial solutions available to help ID lymph > nodes at the grossing table? What are your experiences, pro and con, > of these solutions??? > One of my Pathologist has read a few articles concerning finding lymph nodes > on colon cancer cases. A high number of lymph nodes must be found, > down to > the 1-2 mm size nodes and we are interested in other's experiences. > Thanks for any and all info!!! > 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 > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tom_pella <@t> tedpella.com Tue Jan 4 19:34:34 2005 From: tom_pella <@t> tedpella.com (Tom Pella) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] contract TEM lab In-Reply-To: <000001c4f2ae$2659d7f0$76d48a80@AMY> Message-ID: <00e401c4f2c6$bc3da720$5320fdcc@EMTOM1> I believe the lab announcement was this one, posted in November: Mt Ogden Scientific Services Ogden UT 84401 Tel: 801/334-6677 posted by Connie McManus, who went there to work with her husband. Regards, Tom Pella -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Tuesday, January 04, 2005 2:39 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] contract TEM lab I remember a recent post about a new contract EM lab. I have a client that might need some TEM work done is there anyone out there that does this work for research organizations. Could you e-mail me back with contact information. Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ksamayoa <@t> genomichealth.com Tue Jan 4 19:51:20 2005 From: ksamayoa <@t> genomichealth.com (Kimberly Samayoa) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Histotech Job Opportunity in Northern California Message-ID: <5C9B539E7DD26C4AA41D4E27ACECBB91019BCF4B@jaguar.genomichealth.com> Hi, my name is Kim and I am the only histotech at Genomic Health, a reference lab in Redwood City, CA. We are currently looking for another full time histotech to help us with our commercial operations. Please visit our website at www.genomichealth.com or use this link to our histotech job posting http://www.genomichealth.com/careers/jobs.aspx?JobID=30. Feel free to contact me if you have any questions. Thanks in Advance, Kim Kimberly Brady Samayoa, HT (ASCP) Histotechnologist, Operations Genomic Health, Inc. 301 Penobscot Drive Redwood City, CA 94063 www.genomichealth.com Direct: (650) 569-2218 Lab: (650) 5569300 x323 ksamayoa@genomichealth.com 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 jkiernan <@t> uwo.ca Tue Jan 4 23:30:38 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] The "Stuff" to Find Lymph Nodes References: <8784A3EE34199644AF60DBE517F5B0A60A8A571C@louex04.lou.medcity.net> <000901c4f2b9$564a54c0$63b05bd1@wolf> Message-ID: <41DB7B7E.87F8BF75@uwo.ca> This has been the subject of many Histonet enquiries and answers, the most informed ones being from Dr Bob Richmond, an American pathologist. It's evident that the solutions for revealing lymph nodes are variants of the alcohol-formalin-acetic (AFA) family of fixatives. Bob R (and others) have recommended Davidson's fluid (formalin 40, alcohol 60, acetic acid 20, water 60). Patrick P's formula, cited below, is closer to the traditional early 20th century AFA fixatives (such as those of Tellyesniczky and of Bodian), in that it contains more alcohol and less acetic acid than Davidson's. Bob Richmond has (?had) an excellent web page about Davidson's fixative and its value in lymph node dissection, at http://members.aol.com/RSRICHMOND/histology.html [It took three tries to find this URL, using Google. Two top-of-the-list links failed to find it. Are you still histonetting, Bob? Maybe you should tweak the old page a bit.] If the revelation of lymph nodes is due to partial removal of similarly pale bits of fat, any liquid that can partly dissolve fat should do the job. An obvious answer is 40% ethanol, as in gin or vodka. For a lab poisoned duty-free spirits, diluted 50/50 with tap water, should clarify the lipids and conserve the pallid proteins and DANA of the lymph nodes. John Kiernan London, Canada. _________________________________ Patrick Paulusse wrote: > > Wanda, > > Although I have not been fortunate enough to have tried some of the > commercially available lymph node retrieval solutions, I do use a homemade > solution which was outlined in an article (referenced below) that you might > be interested in trying. The recipe is as follows: > > 500 ml of absolute ethanol > 170 ml distilled water > 80 ml of 37% formaldehyde > 50 ml of acetic acid > > According to the article the reagents must be added in this order (does > anyone know why?). I've used this since 2001 with great success. I dissect > off the pericolic adipose tissue and leave it sit in the solution for 24 > hours and the smallest of lymph nodes show up white against the grey/yellow > adipose tissue. I've also let it sit over the weekend and have seen no > deleterious affects because of the prolonged treatment. If you try it let > me know what you think. > > Article Reference: > > "GEWF solution: an inexpensive, simple, and effectiveaid for the retrieval > of lymph nodes from colorectal cancer specimens". Ken J. Newell, Barry W. > Sawka, Brian F. Rudick, David K. Driman. Arch Pathol Lab Medicine, May > 2001;125:642-645. > > Patrick Paulusse > Histology Supervisor > Pembroke Regional Hospital > Ontario, Canada > > ----- Original Message ----- > From: "Smith Wanda" > To: > Sent: Tuesday, January 04, 2005 3:40 PM > Subject: [Histonet] The "Stuff" to Find Lymph Nodes > > > Dear Histonetters, > > My mind has gone blank...and I need help please! > > What are some of the commercial solutions available to help ID lymph nodes > > at the grossing table? What are your experiences, pro and con, of these > > solutions??? > > One of my Pathologist has read a few articles concerning finding lymph > nodes > > on colon cancer cases. A high number of lymph nodes must be found, down > to > > the 1-2 mm size nodes and we are interested in other's experiences. > > Thanks for any and all info!!! > > 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 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Wed Jan 5 02:19:42 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Sample questions for HT and QIHC References: Message-ID: <005001c4f2ff$50a3a260$af25d445@domainnotset.invalid> According to the ASCP BOR website, the QIHC "can be earned by meeting the eligibility requirements and taking a 50-item examination on your own computer. Candidates who complete the qualification process in immunohistochemistry, including completion of the eligibility requirements and successful completion of the computer examination will receive documentation of their qualification in immunohistochemistry which is valid for five years." http://www.ascp.org/bor/qualification/qihc.asp There is a topic outline available through a link. This outline is dated November 2004. It appears that the exam is now exam questions only, and not a practical anymore. This appears to be a recent change. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Poteete, Jacquie A." To: "'Wayne Holland'" ; Sent: Tuesday, January 04, 2005 10:18 AM Subject: RE: [Histonet] Sample questions for HT and QIHC > My QIHC exam in the fall of 2003 was all "practical projects" and had only > one question. The answer was to be submitted with the slides. > > Jacquie Poteete MT(ASCP)QIHC > Lead Technologist, IHC Lab > Saint Francis Hospital > Tulsa, OK > japoteete@saintfrancis.com > > -----Original Message----- > From: Wayne Holland [mailto:wayneholland1959@msn.com] > Sent: Tuesday, January 04, 2005 9:00 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Sample questions for HT and QIHC > > > Can anyone give me information on how to get sample questions for the HT and > > QIHC examinations? > Thanks so much! > > _________________________________________________________________ > Is your PC infected? Get a FREE online computer virus scan from McAfee(r) > Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ********* Email Confidentiality Statement ********* > Visit http://www.saintfrancis.com/emailconf.asp > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Wed Jan 5 02:35:04 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Sample questions for HT and QIHC References: Message-ID: <005701c4f301$75221020$af25d445@domainnotset.invalid> NSH has Self-Assessment booklets, 14 different topics, each with about 100 multiple choice questions divided into basic, intermediate and advanced levels of questions. The complete fourteen (14) book series is now available on CD-Rom. The cost is $100 for NSH members and $175 for non members. Individual booklets are still available (until supply lasts) at $15 each for NSH members, $35 each for non-members. If you are going for your HT exam, I would suggest the CD-ROM, as it is cheaper to get the CD-ROM, rather than each of the booklets. The 14 topics include: - Fixation - Processing - Embedding/Sectioning - Routine Staining (theory of staining, H&E, Giemsa, MGP, Feulgen) - IHC, ISH, EM - Microorganism staining - Connective tissue staining - Pigment, Mineral and Artifact staining - Nerve and Special Cells staining - Carbohydrate staining - Laboratory Operations (lab math, management, education methodology, equipment, regulations, etc.) - Gross Dissection and Description - Cytopreparation - Laboratory Safety For information about the booklets/CD-ROM: http://www.nsh.org/education/materials.html For information on how to join NSH (you do NOT have to be certified to join)($20 for half-year dues January - May, with renewal in June at $40 for a full year) http://www.nsh.org/membership/application.html (Shameless plug for NSH, I admit it. No profit gained on my part.) Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Wayne Holland" To: Sent: Tuesday, January 04, 2005 10:00 AM Subject: [Histonet] Sample questions for HT and QIHC > Can anyone give me information on how to get sample questions for the HT and > QIHC examinations? > Thanks so much! > > _________________________________________________________________ > Is your PC infected? Get a FREE online computer virus scan from McAfee? > Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From miffordclark <@t> optonline.net Wed Jan 5 04:21:55 2005 From: miffordclark <@t> optonline.net (clifford berger) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] The "Stuff" to Find Lymph Nodes References: <8784A3EE34199644AF60DBE517F5B0A60A8A571C@louex04.lou.medcity.net> <000901c4f2b9$564a54c0$63b05bd1@wolf> Message-ID: <001901c4f310$61ebdbd0$0200a8c0@dellovo0ll7kuk> Hello Wanda, Please take a look at Dissect Aid on our website, www.decal-bone.com. It is a special fixative for revealing lymph nodes which we have been manufacturing for over 20 years. You can call our toll free number or reply to this email for a free sample. Cliff Berger Decal Chemical Corp 1800-428-5856 www.decal-bone.com ----- Original Message ----- From: "Patrick Paulusse" To: "Histonet" ; "Smith Wanda" Sent: Tuesday, January 04, 2005 6:58 PM Subject: Re: [Histonet] The "Stuff" to Find Lymph Nodes > Wanda, > > Although I have not been fortunate enough to have tried some of the > commercially available lymph node retrieval solutions, I do use a homemade > solution which was outlined in an article (referenced below) that you > might > be interested in trying. The recipe is as follows: > > 500 ml of absolute ethanol > 170 ml distilled water > 80 ml of 37% formaldehyde > 50 ml of acetic acid > > According to the article the reagents must be added in this order (does > anyone know why?). I've used this since 2001 with great success. I > dissect > off the pericolic adipose tissue and leave it sit in the solution for 24 > hours and the smallest of lymph nodes show up white against the > grey/yellow > adipose tissue. I've also let it sit over the weekend and have seen no > deleterious affects because of the prolonged treatment. If you try it let > me know what you think. > > Article Reference: > > "GEWF solution: an inexpensive, simple, and effectiveaid for the retrieval > of lymph nodes from colorectal cancer specimens". Ken J. Newell, Barry W. > Sawka, Brian F. Rudick, David K. Driman. Arch Pathol Lab Medicine, May > 2001;125:642-645. > > Patrick Paulusse > Histology Supervisor > Pembroke Regional Hospital > Ontario, Canada > > ----- Original Message ----- > From: "Smith Wanda" > To: > Sent: Tuesday, January 04, 2005 3:40 PM > Subject: [Histonet] The "Stuff" to Find Lymph Nodes > > >> Dear Histonetters, >> My mind has gone blank...and I need help please! >> What are some of the commercial solutions available to help ID lymph >> nodes >> at the grossing table? What are your experiences, pro and con, of these >> solutions??? >> One of my Pathologist has read a few articles concerning finding lymph > nodes >> on colon cancer cases. A high number of lymph nodes must be found, down > to >> the 1-2 mm size nodes and we are interested in other's experiences. >> Thanks for any and all info!!! >> 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 >> > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Lynne.Bell <@t> hitchcock.org Wed Jan 5 06:25:07 2005 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] The "Stuff" to Find Lymph Nodes Message-ID: We use a product called Dissect Aid available from Decal Corp. The website is www.decal-bone.com. The phone number is 1-800-428-5856. The product contains water, glacial acetic acid, ethyl alcohol, formaldehyde. Our pathologists really like it although one of them says if the nodes are in it longer than 24 hours, the H & E sections seem to be a bit pale. Lynne A. Bell, HT (ASCP) Central Vermont Hospital P. O. Box 547 Barre, VT 05641 802-371-4122 From ajennings <@t> unmc.edu Wed Jan 5 07:35:27 2005 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Sample questions for HT and QIHC In-Reply-To: <005701c4f301$75221020$af25d445@domainnotset.invalid> Message-ID: Nice post Peggy I hope you save it somewhere so you don't have to regenerate it again.....I know this question comes up now and again on the Histonet, NSH profit sharing,,,,now there's a thought :-) ijk Anita Sent by: To histonet-bounces@ "Wayne Holland" lists.utsouthwest , ern.edu cc 01/05/2005 02:35 Subject AM Re: [Histonet] Sample questions for HT and QIHC NSH has Self-Assessment booklets, 14 different topics, each with about 100 multiple choice questions divided into basic, intermediate and advanced levels of questions. The complete fourteen (14) book series is now available on CD-Rom. The cost is $100 for NSH members and $175 for non members. Individual booklets are still available (until supply lasts) at $15 each for NSH members, $35 each for non-members. If you are going for your HT exam, I would suggest the CD-ROM, as it is cheaper to get the CD-ROM, rather than each of the booklets. The 14 topics include: - Fixation - Processing - Embedding/Sectioning - Routine Staining (theory of staining, H&E, Giemsa, MGP, Feulgen) - IHC, ISH, EM - Microorganism staining - Connective tissue staining - Pigment, Mineral and Artifact staining - Nerve and Special Cells staining - Carbohydrate staining - Laboratory Operations (lab math, management, education methodology, equipment, regulations, etc.) - Gross Dissection and Description - Cytopreparation - Laboratory Safety For information about the booklets/CD-ROM: http://www.nsh.org/education/materials.html For information on how to join NSH (you do NOT have to be certified to join)($20 for half-year dues January - May, with renewal in June at $40 for a full year) http://www.nsh.org/membership/application.html (Shameless plug for NSH, I admit it. No profit gained on my part.) Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Wayne Holland" To: Sent: Tuesday, January 04, 2005 10:00 AM Subject: [Histonet] Sample questions for HT and QIHC > Can anyone give me information on how to get sample questions for the HT and > QIHC examinations? > Thanks so much! > > _________________________________________________________________ > Is your PC infected? Get a FREE online computer virus scan from McAfee? > Security. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 > > > _______________________________________________ > Histonet mailing list > Histonet@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 histo <@t> bthosp.com Wed Jan 5 08:35:05 2005 From: histo <@t> bthosp.com (O'Brien, Sue) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Regulations regarding placenta retention - when NOT sent to lab Message-ID: <4D95C24EC0E4C84787A6919F1165B25E24B082@btmhems01.BTHOSP.INT> I was wondering if there were any specific regulations regarding how long Maternity must keep placenta's that do not meet our hospital's criteria to be sent to the lab for Pathologist evaluation. I am aware of the CLIAA -88 and CAP regulations, but they apply to specimens submitted to the lab. Thank-you for any insight, website, or information you are willing to share, Susan O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May Court House, NJ From amarusk1 <@t> FAIRVIEW.ORG Wed Jan 5 09:02:08 2005 From: amarusk1 <@t> FAIRVIEW.ORG (ANN MARUSKA) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] C4d Message-ID: Hi histonetters, Is anyone using C4d on paraffin tissue for their transplant specimens as an IHC stain? I am interested in which product you are using and how it has performed for you. Thanks in advance. Ann Ann Maruska Fairview-University Medical Center Mpls. MN 55454 amarusk1@fairview.org 612-273-9119 The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including “protected health information.” If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:ANN MARUSKA N:MARUSKA;ANN TEL;WORK:612-273-9119 ORG:;LAB EMAIL;WORK;PREF;NGW:AMARUSK1@FAIRVIEW.ORG END:VCARD From sa.drew <@t> hosp.wisc.edu Wed Jan 5 09:26:33 2005 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] C4d Message-ID: We are using C4d from Biogenesis(Cat.#2222-8004) at a 1:600 dilution. We pretreat the slides using an EDTA buffer and HIER in BioCare Medical's DeCloaker. Our protocol on the Ventana immunostainer includes an amplification step. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of ANN MARUSKA Sent: Wednesday, January 05, 2005 9:02 AM To: histonet@pathology.swmed.EDU Subject: [Histonet] C4d Hi histonetters, Is anyone using C4d on paraffin tissue for their transplant specimens as an IHC stain? I am interested in which product you are using and how it has performed for you. Thanks in advance. Ann Ann Maruska Fairview-University Medical Center Mpls. MN 55454 amarusk1@fairview.org 612-273-9119 The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including "protected health information." If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> From gcallis <@t> montana.edu Wed Jan 5 10:06:52 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] (no subject) In-Reply-To: References: Message-ID: <6.0.0.22.1.20050105090242.01b33248@gemini.msu.montana.edu> Depending on the kind of staining wanted, I have stained paraffin sections on slides that were stored for years and didn't have any adverse effects. I was not doing immunostaining, just special stains. Caveat: some special stains may be affected by long term storage, one reported is Masson's Trichchrome for collagen. I would also be careful about longer term storage for immunohistochemical stains. Days are probably not a problem. This has been discussed before on Histonet, check the archives for further info. and At 02:57 PM 1/3/2005, you wrote: >Hello everybody, >I was wondering how long paraplast sections that have been >mounted on slides could be left alone after drying before >staining. Is there any reason they couldn't be left for days >or weeks? >Thanks for your advice, >Frank > >_______________________________________________ >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 ander093 <@t> tc.umn.edu Wed Jan 5 10:24:29 2005 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Re: long term storage of slides In-Reply-To: <6.0.0.22.1.20050105090242.01b33248@gemini.msu.montana.edu> References: <6.0.0.22.1.20050105090242.01b33248@gemini.msu.montana.edu> Message-ID: <6.1.2.0.0.20050105102211.02851eb0@ander093.email.umn.edu> Bielschowsky silver stain is also affected by long term storage of slides--talking months not days. I always cut fresh sections for Biel's. At 10:06 AM 1/5/05, Gayle Callis wrote: >Depending on the kind of staining wanted, I have stained paraffin sections >on slides that were stored for years and didn't have any adverse >effects. I was not doing immunostaining, just special stains. > >Caveat: some special stains may be affected by long term storage, one >reported is Masson's Trichchrome for collagen. I would also be careful >about longer term storage for immunohistochemical stains. Days are >probably not a problem. > >This has been discussed before on Histonet, check the archives for further >info. > > and At 02:57 PM 1/3/2005, you wrote: >>Hello everybody, >>I was wondering how long paraplast sections that have been >>mounted on slides could be left alone after drying before >>staining. Is there any reason they couldn't be left for days >>or weeks? >>Thanks for your advice, >>Frank >> >>_______________________________________________ >>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 ftulenko06 <@t> jcu.edu Wed Jan 5 10:39:03 2005 From: ftulenko06 <@t> jcu.edu (ftulenko06@jcu.edu) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Thanks Message-ID: <9c51cd85.b9f95a28.82b3800@mirapoint.jcu.edu> Thanks to everybody who responded to my question. Frank From gcallis <@t> montana.edu Wed Jan 5 10:47:34 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] large cryostat chucks In-Reply-To: <61E9F2400F53D5119CFC00508B44E33B019F5693@khmcexch.uhsi.org > References: <61E9F2400F53D5119CFC00508B44E33B019F5693@khmcexch.uhsi.org> Message-ID: <6.0.0.22.1.20050105092226.01b90ec0@gemini.msu.montana.edu> We have purchased the waffle weave chucks aka specimen disks from Thermo Electron, formerly Shandon, and cut off the stems to match length of stems on the chucks that come with the cryostat in order to fit in the holder ( they cannot be too long!). Did this at home myself with a vice and good hacksaw!! These are also available from Electron Microscopy Sciences, go to their website, histology products. Also, try EMS, At 08:17 AM 1/4/2005, you wrote: >Hi everyone, >I'm looking for large 1 1/2 inch round crystat chucks for my Leica cryostat, >can anyone suggest vendors? > >thanks in advance, >Sue Kapoor, HT (ASCP) >Histology Coordinator >Kenosha Medical Center >Kenosha, WI >262-653-5570 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 sluhisto <@t> yahoo.com Wed Jan 5 11:35:06 2005 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Collagen IV alpha 3,4,5 Message-ID: <20050105173506.10145.qmail@web51007.mail.yahoo.com> Hello All: I am looking for vendors and protocols (paraffin, frozen, IHC, IMF, etc) for collagen IV alpha 3,4,5 for our human renal biopsies. If you have any information that may aid in my search, it would be most appreciated. Happy New Year to all. Susan --------------------------------- Do you Yahoo!? Yahoo! Mail - You care about security. So do we. From mcanshn <@t> med.umich.edu Tue Jan 4 07:36:09 2005 From: mcanshn <@t> med.umich.edu (Nancy McAnsh) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Holding paraplast sections Message-ID: I work in a research lab and many researchers turn in blocks for multiples of dried, unstained sections. I have found that prolonged holding time (over a year) doesn't affect H&E's but longer than three months might affect the more touchy immunohistochemistry antibodies. Nancy G. McAnsh Research Histology and IPOX Lab 3411CCGC 647-3261 ********************************************************** Electronic Mail is not secure, may not be read every day, and should not be used for urgent or sensitive issues. From jkiernan <@t> uwo.ca Wed Jan 5 11:49:31 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] long term storage of specimens References: <41D9D0C9.9020300@umn.edu> Message-ID: <41DC28AB.679972EB@uwo.ca> For many years our department has stored formaldehyde-fixed human brains in 30% alcohol, for later dissection. The fluid level needs to be checked occasionally, but there has never been trouble with moulds, and there is no smell of formaldehyde (something that made neuroanatomy an unloved discipline in earlier times). -- ------------------------------- 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 _______________________________ Colleen Forster wrote: > > Histonetters, > > I have been asked to post a question: > > A museum has tissue samples for teaching stored in 10% formalin. They > are wondering what other solution they might be able to use that would > be less toxic. This would take the place of the formalin for long term > storage. I believe they take the samples out for demonstration so > putting them in a plastic is not an option. > > Any suggestions???? > > Colleen Forster > U of Mn > 612-626-0436 > > -- > Outgoing mail is certified Virus Free. > Checked by AVG Anti-Virus (http://www.grisoft.com). > Version: 7.0.279 / Virus Database: 265.6.7 - Release Date: 12/30/2004 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MTitford <@t> aol.com Wed Jan 5 12:04:20 2005 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] In-situ hybridization/diagnosis question. Message-ID: <77A94318.1D408B22.00762DB1@aol.com> Are labs anywhere using commercially available in-situ hydridization kits to diagnose tumors (Where the tumor may be due to a single abnormality in the DNA)?. Is this going to be the next big "wave" that will follow the increased use of immunohistochemistry? O.K., that was two questions! Thank you Mike Titford USA Pathology Mobile AL USA From JWEEMS <@t> sjha.org Wed Jan 5 12:18:47 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Biofilm Message-ID: <83AACDB0810528418AA106F9AE9B7F7E50782B@sjhaexc02.sjha.org> Is anyone doing any FISH testing for Biofilm? We have a clinician interested and need information. Thanks! j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- 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 Stacy_McLaughlin <@t> cooley-dickinson.org Wed Jan 5 13:20:15 2005 From: Stacy_McLaughlin <@t> cooley-dickinson.org (Stacy McLaughlin) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Plastic specimen containers Message-ID: <3D502BBF5356D31184650090275B750D0346C8C6@mail.cooley-dickinson.org> Hi everyone, I am searching for plastic specimen containers (unfilled). I am mainly interested in sizes varying from 3x4 inches all the way up to containers large enough for brain, or large specimens. The clearer the plastic, the better. (I've checked the archives, but there's not much to go on.) Anyone willing to share their vendors with me? Thanks in advance and Happy New Year! Stacy McLaughlin HT (ASCP) Lead Tech, Histology THIS E-MAIL MESSAGE IS INTENDED ONLY FOR THE USE OF THE INDIVIDUAL OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If the reader of this e-mail message is not the intended recipient, or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this e-mail message is strictly prohibited. If you have received this e-mail message in error, please immediately notify Cooley Dickinson Healthcare at 413-582-2000 and delete or shred the original message and all copies thereof. Thank you. From gcallis <@t> montana.edu Wed Jan 5 14:04:46 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:27 2005 Subject: questions and Re: [Histonet] Biofilm In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E50782B@sjhaexc02.sjha.org> References: <83AACDB0810528418AA106F9AE9B7F7E50782B@sjhaexc02.sjha.org> Message-ID: <6.0.0.22.1.20050105125103.01b23ac8@gemini.msu.montana.edu> Question: when you say biofilm, do you mean in tissues, associated with an implant, tubing, etc, etc and research project? Or is this a diagnostic consideration with clinician asking for contract work on biofilm? Our Center for Biofilm Engineering is doing FISH on tissues that have biofilms on them and you can contact the chairman, Phil Stewart so he can point you to the person in charge of these projects. phil_s@arc.montana.edu. I am not sure if they do contract work in terms of diagnostic work, but Phil may know of someone doing this. He is a nice gentleman and an expert in biofilms with many contacts in this field. At 11:18 AM 1/5/2005, you wrote: >Is anyone doing any FISH testing for Biofilm? We have a clinician >interested and need information. Thanks! j Joyce Weems 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 mari.ann.mailhiot <@t> leica-microsystems.com Wed Jan 5 14:45:33 2005 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] large chucks Message-ID: Hi Gayle We are now making the square waffle weave chucks. It is someting new for us. |-----------------------------------------------+--------------+------------| | Specimen stage, square stainless steel, | 14020139123 | | | small-28 mm, 1 ea | | | |-----------------------------------------------+--------------+------------| | Specimen stage, square stainless steel, | 14020139124 | | | large-36 mm, 1 ea | | | |-----------------------------------------------+--------------+------------| Best Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From gcallis <@t> montana.edu Wed Jan 5 15:09:41 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Square waffle weave cryostat chucks In-Reply-To: References: Message-ID: <6.0.0.22.1.20050105140031.01b228d8@gemini.msu.montana.edu> For those not familiar with now available square waffle weave cryostat chucks, they are superb little inventions!! They do have a stem on them so they fit perfectly in cryostats ie. Leica 1850, 1800 and others that hold onto a stem. You can see them in a publication in J of Histotechnology, written by the pathologist who invented a clever freezing device that improves orientation, and a perfect block for cryowork - published in 2003, early in the year. He has exhibited/demonstrated this instrument at NSH conventions. Always nice to see new things on the market to make our lives easier inside a cryostat! At 01:45 PM 1/5/2005, you wrote: >Hi Gayle > >We are now making the square waffle weave chucks. It is someting new for >us. > >|-----------------------------------------------+--------------+------------| > | Specimen stage, square stainless steel, | > 14020139123 | | > | small-28 mm, 1 > ea | | | > >|-----------------------------------------------+--------------+------------| > | Specimen stage, square stainless steel, | > 14020139124 | | > | large-36 mm, 1 > ea | | | > >|-----------------------------------------------+--------------+------------| > > > > >Best Regards > >Mari Ann Mailhiot BA HT ASCP >Application Specialist >Leica Technical Assistance Center >800 248 0123 x7267 >847 236 3063 fax >mari.ann.mailhiot@leica-microsystems.com >www.leica-microsystems.com > > > >______________________________________________________________________ >This email has been scanned by the MessageLabs Email Security System. >For more information please visit http://www.messagelabs.com/email >______________________________________________________________________ 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 vazquezr <@t> ohsu.edu Wed Jan 5 15:13:41 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:27 2005 Subject: [Histonet] Square waffle weave cryostat chucks Message-ID: Gayle, how much do they cost and do they have a part number? Robyn OHSU >>> Gayle Callis 1/5/2005 1:09:41 PM >>> For those not familiar with now available square waffle weave cryostat chucks, they are superb little inventions!! They do have a stem on them so they fit perfectly in cryostats ie. Leica 1850, 1800 and others that hold onto a stem. You can see them in a publication in J of Histotechnology, written by the pathologist who invented a clever freezing device that improves orientation, and a perfect block for cryowork - published in 2003, early in the year. He has exhibited/demonstrated this instrument at NSH conventions. Always nice to see new things on the market to make our lives easier inside a cryostat! At 01:45 PM 1/5/2005, you wrote: >Hi Gayle > >We are now making the square waffle weave chucks. It is someting new for >us. > >|-----------------------------------------------+--------------+------------| > | Specimen stage, square stainless steel, | > 14020139123 | | > | small-28 mm, 1 > ea | | | > >|-----------------------------------------------+--------------+------------| > | Specimen stage, square stainless steel, | > 14020139124 | | > | large-36 mm, 1 > ea | | | > >|-----------------------------------------------+--------------+------------| > > > > >Best Regards > >Mari Ann Mailhiot BA HT ASCP >Application Specialist >Leica Technical Assistance Center >800 248 0123 x7267 >847 236 3063 fax >mari.ann.mailhiot@leica-microsystems.com >www.leica-microsystems.com > > > >______________________________________________________________________ >This email has been scanned by the MessageLabs Email Security System. >For more information please visit http://www.messagelabs.com/email >______________________________________________________________________ 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 MTitford <@t> aol.com Wed Jan 5 15:18:59 2005 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] In-situ hybridization diagnosis question. Message-ID: <047B6671.11FE013E.00762DB1@aol.com> Are any histology labs anywhere using commercially available in-situ hybridization kits to demonstrate tumors for diagnosis (caused by a single DNA abnormality)? Is this going to be the next "wave" of new techniques in the histology lab, following after immunohistochemistry? O.K., I know, that was two questions! Thank you Mike Titford USA Pathology Mobile AL USA From gcallis <@t> montana.edu Wed Jan 5 16:12:19 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Square waffle weave cryostat chucks In-Reply-To: References: Message-ID: <6.0.0.22.1.20050105151014.01b32b58@gemini.msu.montana.edu> Part number is in the message, have no idea what they cost, get them from Leica Microsystems. You would have to plug the part number into website or contact Leica rep. Gayle Callis At 02:13 PM 1/5/2005, you wrote: >Gayle, >how much do they cost and do they have a part number? > >Robyn >OHSU > > >|-----------------------------------------------+--------------+--------- > ---| > > | Specimen stage, square stainless steel, | ># 14020139123 > > | small-28 mm, 1 > > >|-----------------------------------------------+--------------+--------- > ---| > > | Specimen stage, square stainless steel, | ># 14020139124 | | > > | large-36 mm > > > > > > > > > >Best Regards > > > >Mari Ann Mailhiot BA HT ASCP > >Application Specialist > >Leica Technical Assistance Center > >800 248 0123 x7267 > >847 236 3063 fax > >mari.ann.mailhiot@leica-microsystems.com > >www.leica-microsystems.com > > > > > > > >______________________________________________________________________ > >This email has been scanned by the MessageLabs Email Security System. > >For more information please visit > http://www.messagelabs.com/email > >______________________________________________________________________ > >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 wood <@t> dcpah.msu.edu Wed Jan 5 16:48:07 2005 From: wood <@t> dcpah.msu.edu (Thomas Wood) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Job posting Michigan State University Message-ID: Histology Technician Position at a new building and lab Michigan State University Diagnostic Center for Population and Animal Health Veterinary Medicine Required: knowledge normally acquired in the first two or three years in college, technical or vocational school in histology or chemistry; one year of related and professionally more responsible or expansive histology experience , methods, procedure, terminology, and utilization of special purpose precision instruments; or an equivalent combination of education and experience; certification by the American Society of Clinical Pathologists. Desired: quality microtome sectioning and experience in animal disease recognition, laboratory safety techniques, mixing reagents and other stock solutions, and routine equipment maintenance; computer usage. Basic Function: prepare tissue slides for pathologists, maintain laboratory supplies and equipment; provide histological services (i.e. staining, embedding, trimming, sectioning, cover slipping, labeling of animal tissues, additional special stains and techniques, and assist on research projects); will mix reagents and other stock solutions, dispose of solutions and chemicals, file blocks, and perform daily data entry and other tasks as needed assigned. Work hours will be determined and may very, Currently: (1) shift, no weekends. For an application, visit http://www.hr.msu.edu/, select Hiring/Jog Posting, Support Staff, Forms, or visit us at 1407 S.Harrison, Room 110 Nisbet, East Lansing, MI. Refer to posting C40550. Closing date is January 14, 2005. MSU is an affirmative action/equal opportunity institution. Tom Wood lab supervisor Michigan State University Diagnostic Center for Population and Animal Health East Lansing, MI 48823 From LuckG <@t> empirehealth.org Wed Jan 5 17:11:16 2005 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Regulations regarding placenta retention - when NO T sent to lab Message-ID: Sue, This would not appear to be a lab issue if they are not submitted to lab for a path exam. When this question arose here I sent it on to nursing, Risk Management and the OBGYN committee. Specimen retention requirements for the lab are well defined. If a specimen is not submitted to the lab, in my opinion it should be quickly discarded by the dept of origin. Be careful not to let your lab become a repository for specimens that the physicians want to hold. If the placenta is important enough to hold then it should be submitted for a path exam. If not it should be discarded as "medical waste" with the other disposables that are generated or accumulate during a delivery. Good luck. Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: O'Brien, Sue [mailto:histo@bthosp.com] Sent: Wednesday, January 05, 2005 6:35 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Regulations regarding placenta retention - when NOT sent to lab I was wondering if there were any specific regulations regarding how long Maternity must keep placenta's that do not meet our hospital's criteria to be sent to the lab for Pathologist evaluation. I am aware of the CLIAA -88 and CAP regulations, but they apply to specimens submitted to the lab. Thank-you for any insight, website, or information you are willing to share, Susan O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May Court House, NJ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.reilly <@t> jcu.edu.au Wed Jan 5 17:53:43 2005 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Section Longevity In-Reply-To: <6.0.0.22.1.20050105090242.01b33248@gemini.msu.montana.edu> References: Message-ID: <5.1.0.14.0.20050106093158.00a3c170@mail.jcu.edu.au> Dear All, As Gayle suggested, many stains, eg. Perl's Prussian Blue, H & E, are not affected by long term storage of paraffin sections. Some that have given unsatisfactory results are controls for micro-organisms. I have had old sections of controls for Gram positive and negative bacteria, and controls for leptospira that would not stain properly. Now I cut fresh controls and only keep them for a few days. Regards, Laurie. At 09:06 AM 01/05/05 -0700, Gayle Callis wrote: >Depending on the kind of staining wanted, I have stained paraffin sections >on slides that were stored for years and didn't have any adverse >effects. I was not doing immunostaining, just special stains. > >Caveat: some special stains may be affected by long term storage, one >reported is Masson's Trichchrome for collagen. I would also be careful >about longer term storage for immunohistochemical stains. Days are >probably not a problem. > >This has been discussed before on Histonet, check the archives for further >info. > > and At 02:57 PM 1/3/2005, you wrote: >>Hello everybody, >>I was wondering how long paraplast sections that have been >>mounted on slides could be left alone after drying before >>staining. Is there any reason they couldn't be left for days >>or weeks? >>Thanks for your advice, >>Frank >> >>_______________________________________________ >>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 Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. From robint <@t> mediom.qc.ca Wed Jan 5 18:42:16 2005 From: robint <@t> mediom.qc.ca (Robin Turcotte) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] fixative nerve Message-ID: <002d01c4f388$989526f0$f78ceccf@client> Hi everyone, Is anyone know who is the best fixation for nerveous tissus (peripheral nerves sections ). I am working actually on a research protocol with rabbit that will be finish at the end of the month. thank you Robin From immrstambo <@t> hotmail.com Wed Jan 5 19:14:07 2005 From: immrstambo <@t> hotmail.com (Christine Tambasco) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Ideas for the Illinois Society In-Reply-To: <20050103224806.67306.qmail@web11903.mail.yahoo.com> Message-ID: since more and more histotechs are becoming invovled with the billing process (at least as far as i have seen) how about CPT coding and billing-what every histotech needs to know. Also, immunohistochemistry trouble shooting and tumor panels are always interesting, and lastly - recycling-the pros and cons....Good Luck! Christine Tambasco, HT (ASCP) St. Mary's Hospital, Amsterdam, New York 12010 518-841-7287 >From: eileen dusek >To: histonet >Subject: [Histonet] Ideas for the Illinois Society >Date: Mon, 3 Jan 2005 14:48:06 -0800 (PST) > >Hi everyone, >I hope you all had a great and wonderful Holiday with family and friends. > >I am starting to plan for the 2006 Illinois state meeting. We have continued to have wonderful speakers through out the years. I am trying to come up with new topics. If you were to go your state meeting what discussion or workshop would you be drawn to? >Some thought I have are Basic Staining, Cytology/Histology correlations, FNA's, Plant Pathology, and Veterinary Pathology. >I would love to hear from you, perhaps this will help other states also. > >Thank you > >Eileen C. Dusek >Edward Hospital >Naperville, Ill. > > > >--------------------------------- >Do you Yahoo!? > Dress up your holiday email, Hollywood style. Learn more. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From amoklebu <@t> seattlecca.org Wed Jan 5 19:44:15 2005 From: amoklebu <@t> seattlecca.org (Moklebust, Amanda C) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Warthin-Starry on B-5 fixed tissue Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B948013CF38D@wala01.seattlecca.org> I have been asked to try out a Warthin-Starry protocol previously posted on the histonet (Slides are placed in 1% heated sliver nitrate, then developed in a solution of silver nitrate, hydroquinone, and gelatin). I am only getting the control to stain if I skip our usual pretreatment for mercury pigment. No, the control was not fixed in B-5, but we normally put everything into alcoholic iodine for 4 minutes, water rinse, then 1 minute in sodium thiosulfate, and a final water rinse. I have not been working with B-5 fixed tissue for very long, so I would like to know what other techs can recommend. Do I just continue to avoid alcoholic iodine? Is there a better option for B-5 fixed tissue? Thanks in advance for your assistance. Amanda Moklebust, Seattle Cancer Care Alliance 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 Xorren <@t> aol.com Wed Jan 5 21:48:21 2005 From: Xorren <@t> aol.com (Xorren@aol.com) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Histo/cyto position in NY Message-ID: <65.3c0ac299.2f0e0f05@aol.com> Hello friends, If anyone knows of any positions for HT or cyto processor in NY please email any info to Xorren@aol.com Thanks so much ! Happy cutting From jkiernan <@t> uwo.ca Wed Jan 5 23:48:05 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] fixative nerve References: <002d01c4f388$989526f0$f78ceccf@client> Message-ID: <41DCD115.F12ADC62@uwo.ca> The AFA (alcohol-formalin-acetic) fixatives are probably the best if you need to stain axons in paraffin sections. Ask your boss to recommend a textbook. It's all out there, printed in black on white. John Kiernan London, Canada. ______________________________ Robin Turcotte wrote: > > Hi everyone, > > Is anyone know who is the best fixation for nerveous tissus (peripheral nerves sections ). I am working actually on a research protocol with rabbit that will be finish at the end of the month. > > thank you > > Robin > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akbitting <@t> geisinger.edu Thu Jan 6 07:21:52 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Pathcentre woes Message-ID: Does anyone out there using a Shandon Pathcentre to process their tissues have problems with a lot of dry-outs? I have had this machine ripped apart and put back together numerous times and we have the same problems over and over again. I'm ready to put it on Ebay. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From daniel.eberhard <@t> uni-bielefeld.de Thu Jan 6 08:16:54 2005 From: daniel.eberhard <@t> uni-bielefeld.de (Daniel Eberhard) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] methylation sensitive in situ PCR Message-ID: <41DD4856.4050405@uni-bielefeld.de> Dear Listers, I?m searching for experiences/protocols for an "methylation sensitive in situ PCR" on sections to distinguish a methylated from an unmethylated status of a specific gene without losing spatial information. Has this been done? Any hints are welcome. Thank you Daniel From gcallis <@t> montana.edu Thu Jan 6 10:56:29 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] murine eosinophil immunostaining Message-ID: <6.0.0.22.1.20050106094534.01b003f0@gemini.msu.montana.edu> We are embarking on murine eosinophil staining on FFPE fixed lung (not short fixation time!). I will be doing other searches for available antibodies to detect eosinophils, but would like any recommendations from laboratories successfully doing murine eosinophil IHC. 1. Whether this will work on FFPE tissue, if so, retrieval? 2. Are frozen sections a preference - something I would prefer to do anyway. 3. Antibody source Any other suggestions will be welcome as this is NOT a project that originated in our laboratory so we have no control on fixation time in NBF. 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 Jan 6 11:04:24 2005 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] automated special stains Message-ID: Hi everyone: I would love some input from those who have implemented automated special stains in their lab. Pros and cons, please. Also, I would be happy to hear from vendors who want to give information regarding their machines. The stains that we do most frequently are: B&B Gram Steiner GMS ZNAF B-H Gram PAS Giemsa Thanks! From David.Edmondson <@t> christie-tr.nwest.nhs.uk Thu Jan 6 11:17:20 2005 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Plastic specimen containers Message-ID: VWR International http://www.vwr.com have some, but the buckegt are likely to be opaque polycarbonate, , "Nalgene" is a brand with translucent walls, though not crystal clear. Dave Histology Christie Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Stacy McLaughlin Sent: 05 January 2005 19:20 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Plastic specimen containers Hi everyone, I am searching for plastic specimen containers (unfilled). I am mainly interested in sizes varying from 3x4 inches all the way up to containers large enough for brain, or large specimens. The clearer the plastic, the better. (I've checked the archives, but there's not much to go on.) Anyone willing to share their vendors with me? Thanks in advance and Happy New Year! Stacy McLaughlin HT (ASCP) Lead Tech, Histology THIS E-MAIL MESSAGE IS INTENDED ONLY FOR THE USE OF THE INDIVIDUAL OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN INFORMATION THAT IS PRIVILEGED, CONFIDENTIAL AND EXEMPT FROM DISCLOSURE UNDER APPLICABLE LAW. If the reader of this e-mail message is not the intended recipient, or the employee or agent responsible for delivering the message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this e-mail message is strictly prohibited. If you have received this e-mail message in error, please immediately notify Cooley Dickinson Healthcare at 413-582-2000 and delete or shred the original message and all copies thereof. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ******************************************************************************************************************* This e-mail and any files transmitted with it are confidential and solely for the use of the intended recipient. If you have received this e-mail in error you should not disseminate, distribute or copy it. Please notify the sender immediately and delete this e-mail from your system. ******************************************************************************************************************* From SJones <@t> cvm.tamu.edu Thu Jan 6 11:24:51 2005 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Pathcentre woes Message-ID: Hi Angela, Had a dry-out problem with ours a few years ago. It turned out to be a paraffin temperature sensor that was off. We lowered the paraffin temperature and haven't had the problem since. Let me know if it goes on ebay..... 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 >>> "Angela Bitting" 01/06/05 7:21 AM >>> Does anyone out there using a Shandon Pathcentre to process their tissues have problems with a lot of dry-outs? I have had this machine ripped apart and put back together numerous times and we have the same problems over and over again. I'm ready to put it on Ebay. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rbruggeman <@t> psu.edu Thu Jan 6 12:10:22 2005 From: rbruggeman <@t> psu.edu (Richard Bruggeman) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] c-Myc Control Tissue Message-ID: We are using c-Myc (9E10) from Santa Cruz and were wondering what others have used as a positive control tissue when working up this, or other c-Myc antibodies. The manufacturer shows staining in normal human colon as a control, however we were not that pleased with the results that we obtained in this type of tissue. Any suggestions would be appreciated. Thanks! Richard "Trey" Bruggeman Milton S. Hershey Medical Center Penn State College of Medicine Department of Pathology 500 University Drive Hershey, PA 17033 (717) 531-1044 From asmith <@t> mail.barry.edu Thu Jan 6 12:41:57 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Section Longevity Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3C6F@exchsrv01.barrynet.barry.edu> My experience has been that most dye staining methods are unaffected by the age of the sections. Figures 8 and 9 in Biotechnic and Histochemistry 75:124-131 show fresh staining of paraffin sections cut 30 years ago. 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 Laurie Reilly Sent: Wednesday, January 05, 2005 6:54 PM To: Gayle Callis; ftulenko06@jcu.edu; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Section Longevity Dear All, As Gayle suggested, many stains, eg. Perl's Prussian Blue, H & E, are not affected by long term storage of paraffin sections. Some that have given unsatisfactory results are controls for micro-organisms. I have had old sections of controls for Gram positive and negative bacteria, and controls for leptospira that would not stain properly. Now I cut fresh controls and only keep them for a few days. Regards, Laurie. At 09:06 AM 01/05/05 -0700, Gayle Callis wrote: >Depending on the kind of staining wanted, I have stained paraffin sections >on slides that were stored for years and didn't have any adverse >effects. I was not doing immunostaining, just special stains. > >Caveat: some special stains may be affected by long term storage, one >reported is Masson's Trichchrome for collagen. I would also be careful >about longer term storage for immunohistochemical stains. Days are >probably not a problem. > >This has been discussed before on Histonet, check the archives for further >info. > > and At 02:57 PM 1/3/2005, you wrote: >>Hello everybody, >>I was wondering how long paraplast sections that have been >>mounted on slides could be left alone after drying before >>staining. Is there any reason they couldn't be left for days >>or weeks? >>Thanks for your advice, >>Frank >> >>_______________________________________________ >>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 Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 JosefaNava <@t> texashealth.org Thu Jan 6 13:07:54 2005 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] CDX2 antibody Message-ID: <2C515C1049EAF5459EFD8C9B929078A4194462@phdex03.txhealth.org> Hello, Any suggestion where I can order CDX2 antibody? I used to order this from Biocare and it was discontinued. I appreciate your help. Josie 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 jsirak <@t> aerotek.com Thu Jan 6 14:17:46 2005 From: jsirak <@t> aerotek.com (Sirak, Jennifer) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Histology Opportunities Message-ID: <24B42EF96A5E3442B1AF89D458EFA19F2B26C5@AG00-EXMBX02.allegisgroup.com> Hello, My name is Jennifer Sirak and I am a scientific recruiter in Northern New Jersey. We currently have numerous positions available for histologists. We are looking for people with experience in embedding, staining and microtome. The candidates must be HT eligible and if they are certified it is a plus. A BS degree is also required. If anyone is qualified and interested please contact me at (973)-560-5038. Thank you! Jennifer Sirak Recruiter (973)-560-5038 (973)-560-5091 jsirak@aerotek.com 600 Parsippany Rd. Parsippany NJ, 07054 www.aerotek.com ____________________________________________________________________________________________________ This electronic mail (including any attachments) may contain information that is privileged, confidential, and/or otherwise protected from disclosure to anyone other than its intended recipient(s). Any dissemination or use of this electronic email or its contents (including any attachments) by persons other than the intended recipient(s) is strictly prohibited. If you have received this message in error, please notify us immediately by reply email so that we may correct our internal records. Please then delete the original message (including any attachments) in its entirety. Thank you. From shive003 <@t> umn.edu Thu Jan 6 14:21:50 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Parainfluenza 4 IHC Message-ID: <007101c4f42d$5a944a70$41065486@vdl220FAC> I'm posing a question for someone not on the histonet... Does anyone do IHC staining for Parainfluenza 4? If so, do you take samples from outside your facility? Jan Shivers Univ. of Minnesota Vet Diag Lab shive003@tc.umn.edu From mcauliff <@t> umdnj.edu Thu Jan 6 18:00:42 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] H&E staining on GMA plastic In-Reply-To: <002d01c4f388$989526f0$f78ceccf@client> References: <002d01c4f388$989526f0$f78ceccf@client> Message-ID: <41DDD12A.9020907@umdnj.edu> Dear List A colleague wants to do some H and E staining on zebrafish embryos embedded in JB4+ plastic. Does anyone have a good protocol they can share? Thanks in advance! Geoff -- ********************************************** 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 pruegg <@t> ihctech.net Thu Jan 6 15:38:25 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Collagen IV alpha 3,4,5 In-Reply-To: <20050105173506.10145.qmail@web51007.mail.yahoo.com> Message-ID: <200501062140.j06LeQqI014269@pro12.abac.com> Try the U of Iowa Hybridoma Bank, they have lots of collagens. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histology SLU Sent: Wednesday, January 05, 2005 10:35 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Collagen IV alpha 3,4,5 Hello All: I am looking for vendors and protocols (paraffin, frozen, IHC, IMF, etc) for collagen IV alpha 3,4,5 for our human renal biopsies. If you have any information that may aid in my search, it would be most appreciated. Happy New Year to all. Susan --------------------------------- Do you Yahoo!? Yahoo! Mail - You care about security. So do we. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Jan 6 16:21:02 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] H&E staining on GMA plastic In-Reply-To: <41DDD12A.9020907@umdnj.edu> References: <002d01c4f388$989526f0$f78ceccf@client> <41DDD12A.9020907@umdnj.edu> Message-ID: <6.0.0.22.1.20050106145701.01b18f68@gemini.msu.montana.edu> After sectioning, merely dry the sections at 60C. Gill 3 for 10 min up to 20 min. We never used Harris hematoxylin, only progressive hematoxylins. Other progressive hematoxylins will work, i.e. Richard Allan Hematoxylin 1. Bone requires more time in hematoxylin and less time in eosin/phloxine, this counterstain gives wonderful contrast with GMA sections. Rinse 3 changes distilled water 10 dips each Blue with Scotts tap water substitute 2 minutes Rinse 3 changes distilled water 10 dips AIR DRY SECTIONS!!!! eosin/phloxine for 10 minutes, you may find this shorter time even down to 2 min Rinse very quickly in 95% ethanol Rinse very quickly in 100% ethanol AIR DRY SECTIONS and mount coverslip WITHOUT dipping in xylene. If there any funky wrinkles of plastic close to section, remove carefully with a razor blade, sometimes we had curled lips on edges of sections - this way you have a really flat coverglass. Water and solvents (alcohol and xylenes, etc) soften GMA but do NOT remove it, and will cause really ugly wrinkles and cracked plastic. We learned to coverslip without wetting a slide with solvent. The joy of GMA, hydrating sections is NOT necessary, you can immerse dry sections directly into stains, hence the lovely short protocol. By the way, I have some gems for GMA staining, a manual from an expert from long ago, Sharon van de Velde back in the early 80's and also Nate Brinn - he did some very fine work with this plastic, including enzyme histochemical staining as did Sharon. 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 Evelyn.Flynn <@t> childrens.harvard.edu Thu Jan 6 16:25:50 2005 From: Evelyn.Flynn <@t> childrens.harvard.edu (Flynn, Evelyn) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] H&E staining on GMA plastic Message-ID: Dear Geoff and others, This is the protocol that I use. H&E of JB-4 (Plastic) Sections DO NOT USE XYLENE OR OTHER ORGANIC SOLVENTS 1. Harris Hematoxylin, 12 minutes. 2. Rinse in running tap water until clear. 3. Quick dip in eosin. 4. Rinse in running tap water, 10 minutes 5. Air dry 6. Coverslip with Cytoseal or other mounting medium. Evelyn Flynn Children's Hospital, Boston -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Geoff McAuliffe Sent: Thu 1/6/2005 7:00 PM Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] H&E staining on GMA plastic Dear List A colleague wants to do some H and E staining on zebrafish embryos embedded in JB4+ plastic. Does anyone have a good protocol they can share? Thanks in advance! Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LuckG <@t> empirehealth.org Thu Jan 6 16:32:35 2005 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] H&E staining on GMA plastic Message-ID: Geoff, You're in luck (no pun intended). The cover article of Shandon's 2004 Fall issue of "Lab leader" is devoted to this specific topic. If you don't have access to this publication you can probably get it online from their website www.thermo.com/shandon. Good luck, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org -----Original Message----- From: Geoff McAuliffe [mailto:mcauliff@umdnj.edu] Sent: Thursday, January 06, 2005 4:01 PM Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] H&E staining on GMA plastic Dear List A colleague wants to do some H and E staining on zebrafish embryos embedded in JB4+ plastic. Does anyone have a good protocol they can share? Thanks in advance! Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From weneng2004 <@t> yahoo.com Thu Jan 6 16:55:53 2005 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] tissue dry out of alcohol Message-ID: <20050106225553.70107.qmail@web53402.mail.yahoo.com> Dear histonetters, Today I received some samples in 75% alcohol. They were fixed in 10% formalin before. But some of tubes are open and tissues are dry, alcohol vaporated. What will I get after immunostaining of endothelial cell and H&E staining? Does anybody have this experience before? Were they worthy to give a try? Thanks, Wen --------------------------------- Do you Yahoo!? Yahoo! Mail - Find what you need with new enhanced search. Learn more. From m.wei <@t> biogenex.com Thu Jan 6 17:30:25 2005 From: m.wei <@t> biogenex.com (May Wei) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] CDX2 antibody Message-ID: Dear Josie, you asked for any suggestion where you can order CDX2 antibody. You used to order this from Biocare and it was discontinued. I forwarded Sandy's posted email to you and hope it can help you. Thanks, May Wei, M.Med. BioGenex Laboratories On Fri, 19 Nov 2004 12:52:02 -0700 "Johnson, Sandra J. (MCS)" writes: > We use the Biogenex CDX2 concentrate diluted to 1:100 (with citrate > HIER). It looks fabulous in colon carcinoma(nuclear stain) and > doesn't stain mesothelioma, breast, prostate, or lung cancers. > > Sandy Johnson, HTL > IHC Lead Tech, Histology > Mayo Clinic Scottsdale > > -----Original Message----- > From: marsha r price [mailto:mprice26@juno.com] > Sent: Thursday, November 18, 2004 17:23 PM > To: histonet@pathology.swmed.edu > Subject: [Histonet] RE: 2 questions > Histonetters, > I have 2 different questions for you. > > #1. Is anyone using the CDX2 antibody and whose are you using? > > #2. Is there a published document on the cost of routine histology > per > block from start to finish? > > Thank you. > > Marsha Price > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bettina.hutz <@t> orionpharma.com Fri Jan 7 01:09:33 2005 From: bettina.hutz <@t> orionpharma.com (bettina.hutz@orionpharma.com) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] fixative nerve Message-ID: <243E1A79AF5D89438C70513DBADAA4F01B0F14@sfies-exchange1.orionnet.org> Hello, the best ist 2% glutardialdehyde-2% formaldehyde (1+1) in whole body perfusion, with following plastic embedding (methymetacrylate). I used this for routine neurotoxicity studies at BAYER (Germany). But, it is not the easiest and time/cost intensive. Regards, Bettina Hutz Research Assistant Department of Discovery Biology Orion Corporation, ORION PHARMA Orionintie 1, P.O.Box 65, FIN-02101 Espoo, Finland Tel. +358 10 429 2607, Fax +358 10 429 2924 Email: bettina.hutz@orionpharma.com -----Original Message----- From: Robin Turcotte [mailto:robint@mediom.qc.ca] Sent: Thursday, January 06, 2005 2:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fixative nerve Hi everyone, Is anyone know who is the best fixation for nerveous tissus (peripheral nerves sections ). I am working actually on a research protocol with rabbit that will be finish at the end of the month. thank you Robin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Fri Jan 7 01:51:23 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] RE: murine eosinophil immunostaining Message-ID: <9e124e9dd52a.9dd52a9e124e@amc.uva.nl> Hi Gayle, I have an antibody here against human eosinophils that stains like hell. The datasheet said nothing if other species are stained or not. Considering the huge staining intensity it seems worthwhile to test (with a mouse-on-mouse detection system): anti-Eosinophil Major Basic Protein, mouse monoclonal antibody clone BMK-13 (MonoSan 6008, see: www.monosan.com). The antibody stains cryostat sections, smears and FFPE's (after pepsin pretreatment). Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: c.m.vanderloos@amc.uva.nl ----- Original Message ----- >From Gayle Callis Date Thu, 06 Jan 2005 09:56:29 -0700 To Histonet@lists.utsouthwestern.edu Subject [Histonet] murine eosinophil immunostaining We are embarking on murine eosinophil staining on FFPE fixed lung (not short fixation time!). I will be doing other searches for available antibodies to detect eosinophils, but would like any recommendations from laboratories successfully doing murine eosinophil IHC. 1. Whether this will work on FFPE tissue, if so, retrieval? 2. Are frozen sections a preference - something I would prefer to do anyway. 3. Antibody source Any other suggestions will be welcome as this is NOT a project that originated in our laboratory so we have no control on fixation time in NBF. 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 Lawrence.Brett <@t> luht.scot.nhs.uk Fri Jan 7 04:03:24 2005 From: Lawrence.Brett <@t> luht.scot.nhs.uk (Brett, Lawrence) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] C4d on paraffins Message-ID: <857344E7D8F90240935A288998A89D811E775C@wgh-ex1.luht.scot.nhs.uk> We're using the Quindel ab' (product code A213) with Powervision detection and EDTA/NaOH pH8.0 presssure cooking - (for some reason our usual Tris-EDTA didn't work). Dilution of the primary is 1:200, with 30 minute incubation at room temp. Good results. In my hands, one seems to need a very sensitive detection system for this ab', Powervision or Biogenex Super sensitive polymer kit will do the trick, don't expect decent results with the more popular polymer detection kits. Lawrence Brett Royal Infirmary, 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 Myri37 <@t> aol.com Fri Jan 7 06:42:15 2005 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] oil red in MMA Message-ID: <28DABD7B.20010C80.0005167B@aol.com> Hi dear histonnetters and happy new year 2005 !! does anyone have a protocol of oil red staining in tissu embedded in MMA (mixture of butylmetacrylate and methylmetacrylate)? i would like to stain fats in soft tissu on titane. My sections are 50Um thikness. thank you in advance Myriam From jhofecker <@t> yahoo.com Fri Jan 7 08:55:57 2005 From: jhofecker <@t> yahoo.com (Jennifer Hofecker) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Re: Plastic specimen containers Message-ID: <20050107145557.94878.qmail@web21005.mail.yahoo.com> Stacy, I am sure there are many vendors, but I know that Surgipath has many sizes of containers that you can purchase empty. Several sizes are clear(except tops). They are in the catalog under the prefilled section. www.surgipath.com or (800)225-3035 Hope this helps, Jennifer Date: Wed, 5 Jan 2005 14:20:15 -0500 From: Stacy McLaughlin Subject: [Histonet] Plastic specimen containers To: histonet@lists.utsouthwestern.edu Message-ID: <3D502BBF5356D31184650090275B750D0346C8C6@mail.cooley-dickinson.org> Content-Type: text/plain Hi everyone, I am searching for plastic specimen containers (unfilled). I am mainly interested in sizes varying from 3x4 inches all the way up to containers large enough for brain, or large specimens. The clearer the plastic, the better. (I've checked the archives, but there's not much to go on.) Anyone willing to share their vendors with me? Thanks in advance and Happy New Year! Stacy McLaughlin HT (ASCP) Lead Tech, Histology __________________________________ Do you Yahoo!? Yahoo! Mail - Find what you need with new enhanced search. http://info.mail.yahoo.com/mail_250 From bhewlett <@t> cogeco.ca Fri Jan 7 09:16:18 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] RE: murine eosinophil immunostaining References: <9e124e9dd52a.9dd52a9e124e@amc.uva.nl> Message-ID: <000c01c4f4cb$d6ec5f10$6400a8c0@mainbox> Hi Chris and Gayle, BMK-13 certainly works well in both human and rat, I used it for those purposes about 10 years ago. It's definitely worth a try on mouse. Bryan ----- Original Message ----- From: "C.M. van der Loos" To: Sent: Friday, January 07, 2005 2:51 AM Subject: [Histonet] RE: murine eosinophil immunostaining > Hi Gayle, > > I have an antibody here against human eosinophils that stains like > hell. The datasheet said nothing if other species are stained or not. > Considering the huge staining intensity it seems worthwhile to test > (with a mouse-on-mouse detection system): anti-Eosinophil Major Basic > Protein, mouse monoclonal antibody clone BMK-13 (MonoSan 6008, see: > www.monosan.com). The antibody stains cryostat sections, smears and > FFPE's (after pepsin pretreatment). > > Chris van der Loos, PhD > Dept. of Pathology > Academical Medical Center M2-230 > Meibergdreef 9 > NL-1105 AZ Amsterdam > The Netherlands > > phone: +31 20 5665631 > fax: +31 20 6960389 > e-mail: c.m.vanderloos@amc.uva.nl > > ----- Original Message ----- > >From Gayle Callis > Date Thu, 06 Jan 2005 09:56:29 -0700 > To Histonet@lists.utsouthwestern.edu > Subject [Histonet] murine eosinophil immunostaining > > > We are embarking on murine eosinophil staining on FFPE fixed lung (not > short fixation time!). I will be doing other searches for available > antibodies to detect eosinophils, but would like any recommendations > from > laboratories successfully doing murine eosinophil IHC. > > 1. Whether this will work on FFPE tissue, if so, retrieval? > > 2. Are frozen sections a preference - something I would prefer to do > anyway. > > 3. Antibody source > > Any other suggestions will be welcome as this is NOT a project that > originated in our laboratory so we have no control on fixation time in > NBF. > 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 jkiernan <@t> uwo.ca Fri Jan 7 09:51:49 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] oil red in MMA References: <28DABD7B.20010C80.0005167B@aol.com> Message-ID: <41DEB015.5C93244E@uwo.ca> Embedding in butylmethacrylate and methylmethacrylate involves passage through solvents (alcohol or acetone) and the liquid methacrylate monomers. All these liquids dissolve fat and other hydrophobic lipids, and the sections will not contain material stainable with oil red O. -- ------------------------------- 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 _______________________________ Myri37@aol.com wrote: > > Hi dear histonnetters and happy new year 2005 !! > does anyone have a protocol of oil red staining in tissu embedded in MMA (mixture of butylmetacrylate and methylmetacrylate)? > i would like to stain fats in soft tissu on titane. My sections are 50Um thikness. > thank you in advance > Myriam From SJones <@t> cvm.tamu.edu Fri Jan 7 09:54:35 2005 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] H&E staining on GMA plastic Message-ID: Hi Geoff, Here is the one I use. Sarah H&E Stain for Glycol Methacrylate Sections 1. Gill's Hematoxylin for 30-35 minutes Ref: Barbara F. Matlaga 2. Rinse in distilled water, three changes Ethicon Research Foundation 3. Blue in Scott's Tap Water Substitute, 3 minutes Somerville, NJ 08876 4. Rinse in distilled water, two changes The Bulletin SPEP, Vol. IV, No 2 5. Stain in Eosin-Phloxine for 6-8 minutes June 1976 6. Rinse in two changes of 95% alcohol 7. Rinse in two changes of Abs. alcohol, allow sections to remain in the last Abs. for one minute 8. Clear and mount sections in a synthetic mounting medium Scott's Tap Water Sub. Eosin-Phloxine 1 liter distilled water (0.5% Eosin Y, 0.1% Phloxine in 0.2% acetic acid) 10 gm magnesium sulfate, MgSO4 Add 0.6 ml glacial acetic acid to 500 ml distilled water 2 gm sodium bicarbonate Add 2.5 gm Eosin Y and 0.5 gm Phloxine Mix until dissolved, and filter before use Hematoxylin (half-oxidized from Gill) Add ingredients in order: 365 ml distilled water, 125 ml ethylene glycol, 2 gm anhydrous powdered hematoxylin, 0.2 gm sodium iodate (Na2IO3), weigh out accurately. 40.25 gm aluminum sulfate, Al2(SO4)3*18H2O, 0.5 gm anhydrous citric acid. The stain can be used after 1 hour of mixing. Filter through Whatman #1 filter paper before using for the first time. 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 >>> Geoff McAuliffe 01/06/05 6:00 PM >>> Dear List A colleague wants to do some H and E staining on zebrafish embryos embedded in JB4+ plastic. Does anyone have a good protocol they can share? Thanks in advance! Geoff -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Myri37 <@t> aol.com Fri Jan 7 10:06:16 2005 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] oil red in MMA Message-ID: <67B5CE7A.1229EFDC.0005167B@aol.com> wich resin do you think i have to use for embedding my simples and staining with oil red ? i 've heard of technovit do you konw it is it good ? From jcline <@t> wchsys.org Fri Jan 7 10:12:15 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Stacy McLaughlin HT (ASCP)/specimen containers Message-ID: <000401c4f4d3$a791bf00$1d2a14ac@wchsys.org> I use Tri-State Hospital Supply Corp., 3713 West Street, Landover, Maryland 20785, 1-800-442-1980. I hope this is still the same phone number. I have ordered from them for over 15 years. ***** 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 jhofecker <@t> yahoo.com Fri Jan 7 10:50:28 2005 From: jhofecker <@t> yahoo.com (Jennifer Hofecker) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] calling all muscle folks... PHOS coverslipping Message-ID: <20050107165029.83404.qmail@web21003.mail.yahoo.com> Hello to all muscle pros. I was wondering what you are using to coverslip your amylo-phosphorylase (PHOS) slides? Any alternatives to 85% Karo syrup? Thanks in advance, Jennifer __________________________________ Do you Yahoo!? Yahoo! Mail - Easier than ever with enhanced search. Learn more. http://info.mail.yahoo.com/mail_250 From gcallis <@t> montana.edu Fri Jan 7 11:08:06 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] oil red in MMA In-Reply-To: <28DABD7B.20010C80.0005167B@aol.com> References: <28DABD7B.20010C80.0005167B@aol.com> Message-ID: <6.0.0.22.1.20050107100354.01b51b30@gemini.msu.montana.edu> I doubt this would work as the solvents (alcohols, clearants, monomers are really extracting the lipid, an important factor for making MMA work and polymerize anyway is to NOT have fat in bone. The dye would not penetrate the plastic anyway, you would have to remove the plastic completely and that step would continue to remove the fat from your tissue plus your sections are very thick. If you had post fixed your tissue with osmium tetroxide before processing, you may have had more success in detecting/retaining lipids. , At 05:42 AM 1/7/2005, you wrote: >Hi dear histonnetters and happy new year 2005 !! >does anyone have a protocol of oil red staining in tissu embedded in MMA >(mixture of butylmetacrylate and methylmetacrylate)? >i would like to stain fats in soft tissu on titane. My sections are 50Um >thikness. >thank you in advance >Myriam > >_______________________________________________ >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 jhofecker <@t> yahoo.com Fri Jan 7 11:22:21 2005 From: jhofecker <@t> yahoo.com (Jennifer Hofecker) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] calling all muscle folks... PHOS coverslipping In-Reply-To: <3AADFB88753AD31189C100902786B91C14E6DA1B@hch_nt_exchange.hhsc.ca> Message-ID: <20050107172221.89542.qmail@web21004.mail.yahoo.com> Hi Julia, after applying the iodine and air drying, we use 85% Karo syrup to put a coverslip on the slide. It stays readable for about a weeek, but never gets solid and of course, is not considered permanent. We have a big gooey mess of PHOS slides in a drawer that are unreadable. I may try some glycerine. Thanks for responding, Jennifer --- Celebre Julia wrote: > Hi there... > Just curious, when you do coverslip your > Phosphorylases is this permanent? > I personally don't coverslip mine, I grade them for > the pathologist. But one > of my co-workers would use a few drops of iodine > with glycerine and place a > coverslip on top... > > Julia Celebre MLT > Anatomic Pathology > Hamilton General Hospital > 905-527-0271 ext46145 > email: celebrej@hhsc.ca > > > -----Original Message----- > From: Jennifer Hofecker [mailto:jhofecker@yahoo.com] > Sent: Friday, January 07, 2005 11:50 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] calling all muscle folks... PHOS > coverslipping > > > Hello to all muscle pros. > I was wondering what you are using to coverslip your > amylo-phosphorylase (PHOS) slides? Any alternatives > to 85% Karo syrup? > Thanks in advance, > Jennifer > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - Easier than ever with enhanced search. > Learn more. > http://info.mail.yahoo.com/mail_250 > > _______________________________________________ > 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. > > > __________________________________ Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. http://promotions.yahoo.com/new_mail From gcallis <@t> montana.edu Fri Jan 7 11:28:40 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] alternative resin for oil red O In-Reply-To: <67B5CE7A.1229EFDC.0005167B@aol.com> References: <67B5CE7A.1229EFDC.0005167B@aol.com> Message-ID: <6.0.0.22.1.20050107101121.01b39218@gemini.msu.montana.edu> I have a GMA protocol for lipid oil red O, using a water/monomer gradient. Technovit 7100 is a GMA resin. However, you cannot grind a section in GMA since the water softens the plastic plus safety factors to you, you do not want this plastic on your skin nor breathe vapors either. Microtoming would be difficult if you have metal implant in GMA is really designed for thin sections - we never went thicker than 3 um, with 5 um a rarity as we used GMA for really thin sections. However, you might be successful cutting section with a tungsten carbide blade, however the titanium implant will probably ruin the blade edge, a rather expensive thing to have happen. You would be limited to a thinner section, 50 um is really thick for plastic microtomed sections. I presume when you say "soft tissue on titane", you mean the tissue contains titanium implant? At 09:06 AM 1/7/2005, you wrote: >wich resin do you think i have to use for embedding my simples and >staining with oil red ? i 've heard of technovit do you konw it is it good ? > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From ja.mitchell <@t> hosp.wisc.edu Fri Jan 7 11:40:56 2005 From: ja.mitchell <@t> hosp.wisc.edu (Mitchell Jean A.) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] calling all muscle folks... PHOS coverslipping Message-ID: <583D3E9A1E843445BD54E461D1A2F6F30F83F4B4@uwhis-xchng2.hosp.wisc.edu> Jennifer: After the final drying step of our phosphorylase procedure I coverslip with Cytoseal Mounting Media from Richard Allan. I do not have a problem with fading on my phosphorylase sections. I can forward our procedure to you if you would like to compare it with what you use. Jean Mitchell University of Wisconsin Hospital & Clinics Neuromuscular Laboratory Madison, WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jennifer Hofecker Sent: Friday, January 07, 2005 10:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] calling all muscle folks... PHOS coverslipping Hello to all muscle pros. I was wondering what you are using to coverslip your amylo-phosphorylase (PHOS) slides? Any alternatives to 85% Karo syrup? Thanks in advance, Jennifer __________________________________ Do you Yahoo!? Yahoo! Mail - Easier than ever with enhanced search. Learn more. http://info.mail.yahoo.com/mail_250 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Fri Jan 7 11:45:16 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] calling all muscle folks... PHOS coverslipping Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA257F84B@usca0082k08.labvision.apogent.com> Jennifer, Glycerol/Iodine (4:1) works well. Also, you can take the coverslip off, wash, dry and store. Then if you want to see the stain again, just apply the glycerol iodine and it will show up again. Tim Morken Lab Vision - Neomarkers www.labvision.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jennifer Hofecker Sent: Friday, January 07, 2005 8:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] calling all muscle folks... PHOS coverslipping Hello to all muscle pros. I was wondering what you are using to coverslip your amylo-phosphorylase (PHOS) slides? Any alternatives to 85% Karo syrup? Thanks in advance, Jennifer __________________________________ Do you Yahoo!? Yahoo! Mail - Easier than ever with enhanced search. Learn more. http://info.mail.yahoo.com/mail_250 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GDawson <@t> dynacaremilwaukee.com Fri Jan 7 12:02:47 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] CDX2 antibody Message-ID: The CDX-2 antibody from Biogenex (#MU392-UC) works well on FFPE tissues. Glen Dawson -----Original Message----- From: Nava, Josefa [mailto:JosefaNava@texashealth.org] Sent: Thursday, January 06, 2005 1:08 PM To: 'histonet@pathology.swmed.edu' Subject: [Histonet] CDX2 antibody Hello, Any suggestion where I can order CDX2 antibody? I used to order this from Biocare and it was discontinued. I appreciate your help. Josie The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Jan 7 12:36:35 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] oil red in MMA In-Reply-To: <6.0.0.22.1.20050107100354.01b51b30@gemini.msu.montana.edu> Message-ID: <200501071836.j07IaAnP018758@chip.viawest.net> I agree with Gayle on this. One possible solution might be to try and cut diamond wire sections without any embedding, but the wire does not cut soft tissue unless it is completely supported by something hard. You can superglue the tissue to a wood block and use water in the wire cooling troth, but then you would either have to float stain the sections (I have cut 30 micron thick sections of bone this way) or figure out a way to keep them on a slide for oil red o staining. Frozen sectioning is usually the prefered method for demonstrating fat but cutting frozens on titanium implants even using the best tungsten carbide knife and the tape transfer system would be too difficult, I would think. Oh the impossible things we are asked to do...... Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Friday, January 07, 2005 10:08 AM To: Myri37@aol.com; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] oil red in MMA I doubt this would work as the solvents (alcohols, clearants, monomers are really extracting the lipid, an important factor for making MMA work and polymerize anyway is to NOT have fat in bone. The dye would not penetrate the plastic anyway, you would have to remove the plastic completely and that step would continue to remove the fat from your tissue plus your sections are very thick. If you had post fixed your tissue with osmium tetroxide before processing, you may have had more success in detecting/retaining lipids. , At 05:42 AM 1/7/2005, you wrote: >Hi dear histonnetters and happy new year 2005 !! >does anyone have a protocol of oil red staining in tissu embedded in >MMA (mixture of butylmetacrylate and methylmetacrylate)? >i would like to stain fats in soft tissu on titane. My sections are >50Um thikness. >thank you in advance >Myriam > >_______________________________________________ >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 cfavara <@t> niaid.nih.gov Fri Jan 7 13:20:30 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Myelin basic Protein in mouse Message-ID: I have run a quick search in the archives and found nothing. Anyone with experience with Myelin Basic Protein immunohistochemistry in FFPE murine tissue? Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From cwscouten <@t> myneurolab.com Fri Jan 7 13:41:56 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] oil red in MMA Message-ID: <5784D843593D874C93E9BADCB87342AB102632@tpiserver03.Coretech-holdings.com> I am not familiar with diamond wire cutting. Can you give a reference to learn more about that? Another way to cut soft tissue is with a Vibratome. Superglue the tissue to a pedestal, and cut under water(or media). 50 micron sections is fine, they need to be at least that thick. Mount on slides, or react further as free floating sections with our hisotdipper apparatus. Get back to me if this sounds good but you have further questions. 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 Patsy Ruegg Sent: Friday, January 07, 2005 12:37 PM To: 'Gayle Callis'; Myri37@aol.com; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] oil red in MMA I agree with Gayle on this. One possible solution might be to try and cut diamond wire sections without any embedding, but the wire does not cut soft tissue unless it is completely supported by something hard. You can superglue the tissue to a wood block and use water in the wire cooling troth, but then you would either have to float stain the sections (I have cut 30 micron thick sections of bone this way) or figure out a way to keep them on a slide for oil red o staining. Frozen sectioning is usually the prefered method for demonstrating fat but cutting frozens on titanium implants even using the best tungsten carbide knife and the tape transfer system would be too difficult, I would think. Oh the impossible things we are asked to do...... Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Friday, January 07, 2005 10:08 AM To: Myri37@aol.com; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] oil red in MMA I doubt this would work as the solvents (alcohols, clearants, monomers are really extracting the lipid, an important factor for making MMA work and polymerize anyway is to NOT have fat in bone. The dye would not penetrate the plastic anyway, you would have to remove the plastic completely and that step would continue to remove the fat from your tissue plus your sections are very thick. If you had post fixed your tissue with osmium tetroxide before processing, you may have had more success in detecting/retaining lipids. , At 05:42 AM 1/7/2005, you wrote: >Hi dear histonnetters and happy new year 2005 !! >does anyone have a protocol of oil red staining in tissu embedded in >MMA (mixture of butylmetacrylate and methylmetacrylate)? >i would like to stain fats in soft tissu on titane. My sections are >50Um thikness. >thank you in advance >Myriam > >_______________________________________________ >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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From glorialimetti <@t> yahoo.com Fri Jan 7 14:20:51 2005 From: glorialimetti <@t> yahoo.com (Gloria Limetti) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] EM contracts Message-ID: <20050107202051.98521.qmail@web52403.mail.yahoo.com> I work in the Department of Ophthalmology at the Eye and Ear Institute of the University of Pittsburgh. Does anyone know if there is a lab that will process specimens for elctron microscopy for a fee. I have only a few samples to try and I don't want to order the supplies unless it will be an ongoing project. The fees are also needed for a grant proposal. Whether they charge per specimen, per slide, or from beginning to end, anything would be helpful. Cindy Stone 412-647-9174 stonece@upmc.edu __________________________________ Do you Yahoo!? Yahoo! Mail - Find what you need with new enhanced search. http://info.mail.yahoo.com/mail_250 From koellinr <@t> amgen.com Fri Jan 7 14:43:46 2005 From: koellinr <@t> amgen.com (Koelling, Ray) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Myelin basic Protein in mouse Message-ID: <16834C6DFFA6004C88DE4507FB8AE544018CC3BA@wa-mb4-sea.amgen.com> Cynthia, for a couple of years we've run rat-antiMBP from Serotec (MCA408) on mouse brain fixed 24 hours in NBF. At 1:1000-1:2000 overnight at 4 degrees with no retrieval what-so-ever. Then biotinylated rabbit anti rat and so forth. Ray Raymond Koelling Research Scientist, Pathology Amgen Corp. Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Favara, Cynthia (NIH/NIAID) Sent: Friday, January 07, 2005 11:21 AM To: Histonet Subject: [Histonet] Myelin basic Protein in mouse I have run a quick search in the archives and found nothing. Anyone with experience with Myelin Basic Protein immunohistochemistry in FFPE murine tissue? Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ekh9535 <@t> bjc.org Fri Jan 7 14:44:35 2005 From: ekh9535 <@t> bjc.org (Erin Herter) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] ASP 300 Message-ID: Does anyone have a Leica ASP 300 and if so do you have a lot of problems with it? Erin- From gcallis <@t> montana.edu Fri Jan 7 15:55:12 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Diamond wire saws In-Reply-To: <5784D843593D874C93E9BADCB87342AB102632@tpiserver03.Coretec h-holdings.com> References: <5784D843593D874C93E9BADCB87342AB102632@tpiserver03.Coretech-holdings.com> Message-ID: <6.0.0.22.1.20050107145214.01b0caa0@gemini.msu.montana.edu> Diamond wire saws, a wire with teeth in it that cuts bone, and bone embedded in methyl methacrylate plastic usually with precise cutting. Delaware Diamond Knives sells the saw if you want to see it. The lady who needed the Oill red O stain had her tissue, probably bone embedded in methyl methacrylate, very hard and not in soft tissue state and I believe containing a titanium implant. 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 Dixon.Leslie <@t> mayo.edu Fri Jan 7 16:03:15 2005 From: Dixon.Leslie <@t> mayo.edu (Dixon, Leslie E.) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] NBT/BCIP Message-ID: <115ED45FBA6D3D4B89E7F40A9EB5339D01826CD2@excsrv60.mayo.edu> Good afternoon, I have searched the histonet and found NBT/BCIP used to stain for osteoblasts. Does anyone have a protocol for this? Your help is always appreciated. Thanks, Leslie From Myri37 <@t> aol.com Fri Jan 7 17:59:10 2005 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] alternative resin for oil red O Message-ID: <8b.1e395801.2f107c4e@aol.com> the tissue is thin and soft, around a titane implant. i am cutting sections with diamond saw (200 um) and grinding them to about 50 um thikness because of titane i cant grind more than this. Thank you myriam From jkiernan <@t> uwo.ca Sat Jan 8 00:56:00 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] NBT/BCIP References: <115ED45FBA6D3D4B89E7F40A9EB5339D01826CD2@excsrv60.mayo.edu> Message-ID: <41DF8400.D81E1630@uwo.ca> Dear Leslie, The abbreviations you quote probably are: NBT - Nitro blue tetrazolium BCIP - 4-bromo-5-chloro-indolyl phosphate BCIP is a substrate for phosphatase enzymes. It yields, on hydrolysis, phosphate ions and 4-bromo-5-chloro-indoxyl. The latter is a very reactive compound. It can reduce NBT (soluble and yellow) to an insoluble blue formazan at sites of enzymatic activity. (In related methods indoxyls released from BCIP and other esters can generate insoluble indigoid and azoic pigments. The choice of method is often determined by an enzyme's pH optimum.) The "NBT/BCIP" method needed for your purposes will depend on the questions being asked. There are at least three kinds of phosphatase in bone. The tartrate-resistant acid phosphatase (TRAP), typical of of osteoclasts, does occur also in some osteoblasts and bone lining cells (see Nakano et al 2004 J Histochem Cytochem 52:1475-1482; this very recent paper has references to earlier, more important work in the field). There have been many Histonet questions and answers about TRAP. It's a popular enzyme, but is it the subject of your research? In enzyme activity histochemistry there is no such thing as "Does anyone have a protocol?" These methods yield meaningful result only when used with full understanding of how they work. Various control procedures (notably incubations that include specific inhibitors) are also needed in enzyme activity histochemistry. Histonet helps, but it is not enough. If you are doing research there's no easy way to avoid lots of hours in the library. Peer-reviewed papers in good journals are the only sources you can trust. John Kiernan London, Canada. ___________________________________ "Dixon, Leslie E." wrote: > > Good afternoon, > > I have searched the histonet and found NBT/BCIP used to stain for osteoblasts. Does anyone have a protocol for this? Your help is always appreciated. > > Thanks, > Leslie > ---- From pruegg <@t> ihctech.net Sat Jan 8 09:58:06 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] apoptosis question Message-ID: <200501081600.j08G06aC093881@pro12.abac.com> I have been doing cleaved caspase 3 from Cell Signaling with great success on rat samples but one of my customers wants to use it on sheep cells and we are not getting any staining. Has anyone else gotten apoptosis signals from sheep cells? Patsy The dilemma: 1. i have never seen caspase 3 (cleaved or other) by IHC in the sheep cells. 2. im not sure if they have it or the IHC ab doesnt detect it. - So i cant draw a conclusion - 3. with dapi the nuclei look generally okay, as well as by tunel stain on whole mount lungs 4 - what can i do without repeating expt on existing slides to confirm or deny presence of apoptosis (on these sheep cells)??? Annexin V , tunel. apotag? From pruegg <@t> ihctech.net Sat Jan 8 10:07:00 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] alternative resin for oil red O In-Reply-To: <8b.1e395801.2f107c4e@aol.com> Message-ID: <200501081610.j08GA1nW097950@pro12.abac.com> With a diamond wire saw you can cut sections near 30 microns thick, thinner than with a diamond disc saw in my experience. I use a Diamond Wire Saw from Delaware Diamond Knives. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Myri37@aol.com Sent: Friday, January 07, 2005 4:59 PM To: gcallis@montana.edu; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] alternative resin for oil red O the tissue is thin and soft, around a titane implant. i am cutting sections with diamond saw (200 um) and grinding them to about 50 um thikness because of titane i cant grind more than this. Thank you myriam _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Andrew.MacDuff <@t> ed.ac.uk Sun Jan 9 06:37:03 2005 From: Andrew.MacDuff <@t> ed.ac.uk (Andrew MacDuff) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Mouse lung fixation Message-ID: <002401c4f647$ec079f90$b9b2d781@universi8lynpq> Dear All I am just starting a research project and need to fix mouse lungs for embedding in parafin so that I can use them for histology and immunohistochemistry. Could you please tell me the best way to fix the lungs? So far I have tried removing the lungs and then instilling 10% neutral buffered formalin into the trachea with a syringe. This seems to work but I'm not sure how reproducible the alveolar architecture will be. Also do people exsanguinate the mouse before fixing the tissues? (we've been using cervical dislocation and then just removing the lungs) Many thanks Andrew MacDuff Clinical Research Fellow Wilkie Laboratory Medical School Edinburgh University Teviot Place Edinburgh andrew.macduff@ed.ac.uk From RCHIOVETTI <@t> aol.com Sun Jan 9 13:44:33 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Mouse lung fixation Message-ID: In a message dated 1/9/2005 5:44:15 AM US Mountain Standard Time, Andrew.MacDuff@ed.ac.uk writes: am just starting a research project and need to fix mouse lungs for embedding in parafin so that I can use them for histology and immunohistochemistry. Could you please tell me the best way to fix the lungs? So far I have tried removing the lungs and then instilling 10% neutral buffered formalin into the trachea with a syringe. This seems to work but I'm not sure how reproducible the alveolar architecture will be. Also do people exsanguinate the mouse before fixing the tissues? (we've been using cervical dislocation and then just removing the lungs) Andrew, I haven't done lung tissue for paraffin embedding, but I've done a lot of mouse lung prep for EM (using glutaraldehyde instead of formalin). I would recommend perfusion fixation. It's good enough for EM, so it should work great for light microscopy. Contact me off-list if you need additional details. In a nutshell, our protocols usually went something like this: 1. We used chemical anesthesia via intraperitoneal injection, but cervical dislocation should be OK as long as you can get into the chest cavity while the heart is still beating. 2. Use a 26 ga. needle and an IV tubing set with a "piggyback" port on it. I like the small butterfly needles like the ones that are used for pediatric scalp veins...gives you something to hold onto when you go in with the needle. 3. Pre-fix washing solution: Ringer's saline + 0.5% procaine + saccharose to adjust osmolarity to about 350 mOsm (the procaine relaxes the circulatory system's musculature and ensures good access to the lung's capillary bed and the alveoli). Warm to 37 degrees C and hang in an IV bag about 1 Meter above the animal. 4. Remove the sternum and the anterior portions of the ribs in one piece to expose the heart. 5. Move quickly from this point on. Insert the needle through the heart wall into the right ventricle. 6. Start the flow of the wash solution and at the same time make a nick in the left atrium. This gives you a direct route to the pulmonary circulation. 7. Perfuse for about 1 to 1-1/2 minutes. You will know everything's working OK if you see the lungs blanch from pink to white. 8. "piggyback" the fixative solution (NBF?), also warmed to 37 degrees and hanging in another IV bag / tubing setup, into the line that's in the heart. Shut off the washing buffer and start the fixative flowing. 9. Perfusion fix for about 10 minutes in-situ. After that you can remove the lungs and dissect them in more fixative. 10. For EM studies the fixative solution was also adjusted to 380 mOsm with saccharose. I don't know whether this is absolutely necessary for histology, though. Good luck, hope this helps! Cheers, Bob Chiovetti From dbpiontek <@t> clinomicsbio.com Sun Jan 9 14:25:48 2005 From: dbpiontek <@t> clinomicsbio.com (Denise Bland-Piontek) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Job Opportunity in New York Message-ID: <145F33B84793CF488578F86355B32E691704B2@mail.clinomicsbio.com> Lead Histotechnologist Clinomics Biosciences, Inc. is seeking a lead histotechnologist for operation of our Histopathology Laboratory. The candidate will be expected to oversee IHC (many antibody characterizations), TMA (paraffin and frozen) and daily duties for a staff of five. HT or HTL certification required with five plus years of experience. CAP accredited laboratory, maintenance for inspections. Knowledge of in situ hybridization, immunofluoresence and TUNEL a plus. Laser Capture Microdissection is performed in the laboratory. CoolScope is utilized for client interaction and digital library. This is a great opportunity to step outside the box and be a direct participant in a research role. Extremely competitive salary. Clinomics Biosciences is located in Watervliet, NY on Rensselaer Polytechnic Institute campus just outside of Albany, NY. Please contact Denise Bland-Piontek, dbpiontek@clinomicsbio.com (617) 938-1250 for application or questions. Please visit our website www.clinomicsbio.com . Denise Bland-Piontek, HTL(ASCP)CTBS(AATB) Clinomics Biosciences, Inc. Histopathology Manager (617) 938-1250 dbpiontek@clinomicsbio.com From Kathy.Paton <@t> waitematadhb.govt.nz Sun Jan 9 14:47:31 2005 From: Kathy.Paton <@t> waitematadhb.govt.nz (Kathy Paton (WDHB)) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] ASP 300 Message-ID: Hmmmmm....sounds familiar, I purchased the ASP 300 two years ago and yes we did have a few problems with the software. I assume it is software problems you are having. After a few upgrades and the persistent support from our New Zealand agents (Global Science), we have overcome these problems. Some were just irritating flaws in the program, but some of the software problems were quite serious. I am confidant if you contact your agents to arrange for the latest upgrade your problems could be solved. Kathy Paton Surgical Pathology Unit North Shore Hospital Auckland New Zealand -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Erin Herter Sent: Saturday, 8 January 2005 09:45 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] ASP 300 Does anyone have a Leica ASP 300 and if so do you have a lot of problems with it? Erin- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akbitting <@t> geisinger.edu Sun Jan 9 14:56:43 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] I've been trying to stain some brain sections using the Sevier-Munger method for an autopsy case and Message-ID: I've been trying to stain some brain sections using the Sevier-Munger method for an autopsy case and they keep floating off the slide. I used Snow-coat extra slides and dried the sections for 5 days and they still floated. Any ideas? I guess glued slides wouldn't work. Has anyone tried it? Thanks in advance. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From Myri37 <@t> aol.com Mon Jan 10 06:16:26 2005 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Re : alternative resin for oil red O Message-ID: <33D0A9F7.5807B215.0005167B@aol.com> Hi thank you very much for your help. could you please send me your GMA protocol for lipid oil red ? is this resin technovit 7100 easy to do ? or could i buy it already prepared ? i would like to try embedding titane implant with. do you think a postfixation with osmium would be better before embedding with technovit 7100 ? myriam From nancy.troiano <@t> yale.edu Mon Jan 10 07:12:18 2005 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Oil red O on GMA sections Message-ID: <5.2.1.1.2.20050110081039.00b54040@email.med.yale.edu> Gayle and others - did you mention that you had a GMA technique using Technovit 7100 for Oil Red O on undecal bone? If so, can you post it? Thanks so much. Nancy From mward <@t> wfubmc.edu Mon Jan 10 07:43:17 2005 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] CD2 Message-ID: <61135F0455D33347B5AAE209B903A3040BB28671@EXCHVS2.medctr.ad.wfubmc.edu> I have had a request for this antibody and I wondered if anyone is doing this on FFPE and if so what vendor do you use? Thanks in advance for your help. Martha Ward, MT (ASCP) QIHC Wake Forest University Baptist Medical Center From GDawson <@t> dynacaremilwaukee.com Mon Jan 10 08:26:00 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] CD2 Message-ID: Martha, The CD2 from Novocastra (Cat. # NCL-CD2-271)works well on FFPE tissues. Good Luck, Glen Dawson -----Original Message----- From: Martha Ward [mailto:mward@wfubmc.edu] Sent: Monday, January 10, 2005 7:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD2 I have had a request for this antibody and I wondered if anyone is doing this on FFPE and if so what vendor do you use? Thanks in advance for your help. Martha Ward, MT (ASCP) QIHC Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SAllen <@t> exchange.hsc.mb.ca Mon Jan 10 09:42:36 2005 From: SAllen <@t> exchange.hsc.mb.ca (Sharon Allen) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] calling all muscle folks... PHOS coverslipping Message-ID: <304B5A264EC9974E8121B0EA14A9C3936619D6@hsc01mx1.hsc.mb.ca> Hi, We use glycerine with a few drops of iodine, then rim the coverslip with clear nail polish. This works well & the slides can be filed with the rest. Hope this helps. Sharon salle@hsc.mb.ca -----Original Message----- This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. From gcallis <@t> montana.edu Mon Jan 10 10:43:33 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] murine eosinophil immunostaining In-Reply-To: <200501081600.j08G06aB093881@pro12.abac.com> References: <6.0.0.22.1.20050106094534.01b003f0@gemini.msu.montana.edu> <200501081600.j08G06aB093881@pro12.abac.com> Message-ID: <6.0.0.22.1.20050110093648.01b5af80@gemini.msu.montana.edu> Dear Patsy, The group specifically requested immunostaining which is being abandoned in favor of Bryan Llewwllyn's siruius red, compliments of Ian Montgomery and does stain murine eosinophils very well. I have seen murine eo's in H&E but I am deferring to a more specific request and certainly a lot cheaper in terms of time and money than enumerating murine eosinophils in 6 lung samples using IHC!! I saw a great deal of discussion on murine eosinophils in the HISTONET archives, with the same answers I have collected. It turns out that there is no specific murine eosinophil antibody at this time, something we are looking for and we may try the MONOSAN (aka available through Cedarlane and/or Accurate in USA), murine antihuman BMK13 in the future. At this time IHC is on hold in favor of saving big bucks and time. At 08:58 AM 1/8/2005, you wrote: >Gayle have you tried just ordinary H&E or wright/giemsa type stains which >usually stain eosinophils nicely in my hands, even murine >Patsy > >-----Original Message----- > >Subject: [Histonet] murine eosinophil immunostaining > >We are embarking on murine eosinophil staining on FFPE fixed lung (not >short fixation time!). I will be doing other searches for available >antibodies to detect eosinophils, but would like any recommendations from >laboratories successfully doing murine eosinophil IHC. > >1. Whether this will work on FFPE tissue, if so, retrieval? > >2. Are frozen sections a preference - something I would prefer to do >anyway. > >3. Antibody source > >Any other suggestions will be welcome as this is NOT a project that >originated in our laboratory so we have no control on fixation time in NBF. >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Jan 10 11:09:06 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Eosinophil Major Basic Protein MON6008 does not cross react to mouse Message-ID: <6.0.0.22.1.20050110100504.01b38ca0@gemini.msu.montana.edu> After doing all the search for a murine eosinophil antibody, MONOSAN on antibody suggested by Chris van der Loos replied with following message. It does not cross react to murine eosinophils. Hopefully this saves someone some time and effort. Gayle Callis >Subject: Eosinophil Major Basic Protein >Date: Mon, 10 Jan 2005 16:23:25 +0100 > >From: "Joreen Huijbregts" > >Dear Mrs. Callis, > >Thank you for visiting our website. In addition to your request about >the Human Eosinophil Major Basic Protein we can inform you that the >antibody MON6008 does not cross-react with mouse. > >You can purchase our antibodies in the US from two distributors: > >Cell Sciences, www.cellsciences.com >Caltag Laboratories Inc, www.caltag.com > >If you have any additional questions or remarks, please do not hesitate >to contact us or our distributors. > >Kind regards, >S A N B I O b.v. > >Ms. Joreen Huijbregts >Monosan Technical Support >jhuijbregts@sanbio.nl >Tel. +31 413 251115 >Fax. +31 413 266605 > >www.monosan.com > From gcallis <@t> montana.edu Mon Jan 10 11:35:03 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Oil red O on GMA sections In-Reply-To: <5.2.1.1.2.20050110081039.00b54040@email.med.yale.edu> References: <5.2.1.1.2.20050110081039.00b54040@email.med.yale.edu> Message-ID: <6.0.0.22.1.20050110101137.01b83b00@gemini.msu.montana.edu> Dear all This is NOT an oil red O for undecalcified bone in GMA, but a general Oil red O for lipids in soft tissues that are NBF fixed. Technovit 7100 is a kit and I have not used it but I have cut tissues embedded in this GMA, it is superb and a bit pricey. Personally, I do not do like GMA embedded undecalcified bone, it has many inherent problems one of which can be problems with controlling polymerization along with poor infiltration plus being difficult to section. GMA is just plain SOFTER than MMA! Nancy, if I were to embark on a project like this, I would prefer an osmium tetroxide post fixation, then embed into MMA and section. The lipid will be black, and remains in the tissue during processing and embedding as it does with epoxy resins for EM. First, with any GMA for lipid, you cannot do any dehydration in alcohols, you would have to do a monomer gradient (expensive with a kit like Technovit 7100, archives will say where that can be purchased from) using decreasing amounts of water. Sharon van de Velde simple uesd Monomer A from the JB4 kit, PolySciences or the monomer from the Technovits kit. You need to do extra changes to remove all the water. Using the monomer from a Technovits kit could be very expensive unless you buy extra for this special processing. Remember that GMA polymerization is very difficult to control in a thick bone sample, you may be limited to the usual 1 -2 mm thickness with cortical bone, although it works well with Jamshidi needle biopsies. The oil red O protocol is the standard Oil red O found in histology text books, using propylene glycol. Unfortunately, I do not have this on computer file but I do have the Churukian oil red o, I will attach that to you personally. You may have to look it up or it should be available in Histonet archives, many people have responded with their recipes in past. I know the standard propylene glycol method, how to make up stain is found in some website links found on Histosearch website. Staining is for GMA sections (2 um thick!) with standard Oil red O in propylene glycol, a messy method! absolute propylene glycol for 2 min oil red O for 2 hours differentiate in 85% propylene glycol 1 - 2 min Hematoxylin counterstain Mount with aqueous mounting media I suggest you try the Churukian Oil red O stain, much nicer than messy propylene glycol but you would have to work out staining time. 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 BetStark <@t> aol.com Mon Jan 10 11:36:43 2005 From: BetStark <@t> aol.com (BetStark@aol.com) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] histo position northwestern Ct. Message-ID: <97.56882cae.2f14172b@aol.com> Northwest CT. 90 minutes north of New York City Sharon Hospital, located in the picturesque corner of northwest Connecticut is a 78 bed acute care facility and is currently seeking a histology tech ASCP certified or eligible. General histology duties, including embedding, cutting, special stains, maintenance of instruments and CAP, JCAHO, CLIA compliance. A great lab in a beautiful small, New England town. After 26 years, I retired leaving many friends and fond memories. Sharon Hospital provides its employees with an outstanding wage and benefits package. To apply, please mail/fax.email your resume to: Human Resources Sharon Hospital 50 Hospital Hill Rd. Sharon, CT. 06069 Fax: 860-364-4465 Email: hr@sharonhospital.com . From carolb <@t> mail.phys.mcw.edu Mon Jan 10 12:46:20 2005 From: carolb <@t> mail.phys.mcw.edu (Carol Bobrowitz) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] paraplast - coverslips Message-ID: <59DF6640CDC0D7119DF000D0B761393C1A961A@thor.phys.mcw.edu> When doing insitu I used a coverslip made from silicone or paraplast during the incubation / hybridization step. I had several different sizes. I can't recall the Vendor's name. Does anybody have a Vendor who sells these? Thank you, Carol Ann Bobrowitz Histology Laboratory Department of Physiology - Room 541 Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, Wisconsin 53226 414-456-8179 FAX 414-456-6546 From BWinters <@t> NCH.ORG Mon Jan 10 12:59:08 2005 From: BWinters <@t> NCH.ORG (Winters, Bert) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] CALIBRATING SCALES Message-ID: <270614B321ACB44D8C1D91F4F921FDC3627790@NCH01EX02.nch.org> We have a new balance that needs to be calibrated by our biomed. department before we can put it in service. The weights that we having using for years are no longer the correct standard for calibrating the scale and biomed tells us we need to order special weight that will exceed $1,000. Does anyone know of a service that will come out and calibrate the scale. That seems to be a more cost efficient way of taking care of this problem. Any advice would be greatly appreciated. Bert Winters, Northwest community hospital BWINTERS@NCH.ORG ******************* PLEASE NOTE ******************* This E-Mail/telefax message and any documents accompanying this transmission may contain information that is privileged, confidential, and/or exempt from disclosure under applicable law and is intended solely for the addressee(s) named above. If you are not the intended addressee/recipient, you are hereby notified that any use of, disclosure, copying, distribution, or reliance on the contents of this E-Mail/telefax information is strictly prohibited and may result in legal action against you. Please reply to the sender advising of the error in transmission and immediately delete/destroy the message and any accompanying documents. Thank you. From kmerriam2003 <@t> yahoo.com Mon Jan 10 13:18:26 2005 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] c-fos antibody in mouse FFPE tissues Message-ID: <20050110191827.5893.qmail@web52503.mail.yahoo.com> Hello, Does anyone know what would be the best positive control tissue for this antibody in mouse tissue? Thanks in advance, Kim Merriam Novartis Cambridge, MA Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? The all-new My Yahoo! – Get yours free! From rlprocto <@t> email.unc.edu Mon Jan 10 14:31:48 2005 From: rlprocto <@t> email.unc.edu (rlprocto@email.unc.edu) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Sections floating off of slides Message-ID: <20050110153148.fv63ghmn0ggsssos@webmail3.isis.unc.edu> Hello All, I have encountered a problem H&E staining my paraffin embedded 8um mouse brain sections. The sections are floating right off some of the slides but some of the samples are normal. The sectioning was done between mid-November 2004 through last week. The ones sectioned more recently were embedded simultaneously and seem to be fine. I tried baking the slides at 55 C for an hour, the problem persisted, but at a slower rate. Any tips to correct this problem would be greatly appreciated. Becky Proctor UNC-Chapel Hill From liz <@t> premierlab.com Mon Jan 10 14:59:47 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] CD21, B220, CD4 Message-ID: <000201c4f757$51e16d40$76d48a80@AMY> I need to run these antibodies on mouse tissue. Can anyone recommend me a good source and if they might possibly work in FFPE tissue or do I need to use frozens. Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From mcauliff <@t> umdnj.edu Mon Jan 10 18:16:40 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] CALIBRATING SCALES In-Reply-To: <270614B321ACB44D8C1D91F4F921FDC3627790@NCH01EX02.nch.org> References: <270614B321ACB44D8C1D91F4F921FDC3627790@NCH01EX02.nch.org> Message-ID: <41E31AE8.7020301@umdnj.edu> Hi Brian: I must admit that I was skeptical until I looked in the Fischer catalogue, a complete set of weights lists for $980. That said, what is the matter with your "old" weights? Maybe if you need great accuracy to 4 decimal places you might need some very precise weights, but otherwise I think not. What sort of reagent needs that degree of precision? When we had Mettler come out and calibrate out balances the technician always had his own set of weights. I would talk to the manufacturer, the vendor and whoever is providing service. Geoff Winters, Bert wrote: >We have a new balance that needs to be calibrated by our biomed. department before we can put it in service. The weights that we having using for years are no longer the correct standard for calibrating the scale and biomed tells us we need to order special weight that will exceed $1,000. Does anyone know of a service that will come out and calibrate the scale. That seems to be a more cost efficient way of taking care of this problem. Any advice would be greatly appreciated. > > > Bert Winters, Northwest community hospital > > BWINTERS@NCH.ORG > > >******************* PLEASE NOTE ******************* >This E-Mail/telefax message and any documents accompanying this transmission may contain information that is privileged, confidential, and/or exempt from disclosure under applicable law and is intended solely for the addressee(s) named above. If you are not the intended addressee/recipient, you are hereby notified that any use of, disclosure, copying, distribution, or reliance on the contents of this E-Mail/telefax information is strictly prohibited and may result in legal action against you. Please reply to the sender advising of the error in transmission and immediately delete/destroy the message and any accompanying documents. Thank you. >_______________________________________________ >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 gcallis <@t> montana.edu Mon Jan 10 15:46:23 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Sections floating off of slides In-Reply-To: <20050110153148.fv63ghmn0ggsssos@webmail3.isis.unc.edu> References: <20050110153148.fv63ghmn0ggsssos@webmail3.isis.unc.edu> Message-ID: <6.0.0.22.1.20050110143825.01b0c418@gemini.msu.montana.edu> Several things could be happening 1. No adhesive in either the waterbath - some add a few granules of gelatin as waterbath heats up or not using Plus Charge (silane or poly l lysine coated) slides to keep sections adhered to the slide surface. If you use Plus charge slides, the water should be distilled and not tap water as minerals in tap water in certain parts of the country will attract to the Plus charge and negate section adherance. 2. If you use ammonia water to blue the hematoxylin, it may be chewing the protein sections off the slides aka alkaline protein hydrolysis which makes sections release. Change to Scotts tap water substitute to blue you sections. This problem can occure even after oven drying of sections. If you use Ammonia water, it should be very dilute, just enough to blue the section, change it daily since it is a cheap solution. Sections can release with ammonia water bluing even IF you use slide adhesives. 3. Hopefully, your brain sections are very flat to perfect contact with the slide. Sharp blades are a must. At 01:31 PM 1/10/2005, you wrote: >Hello All, >I have encountered a problem H&E staining my paraffin embedded 8um mouse brain >sections. The sections are floating right off some of the slides but some of >the samples are normal. >The sectioning was done between mid-November 2004 through last week. The ones >sectioned more recently were embedded simultaneously and seem to be fine. >I tried baking the slides at 55 C for an hour, the problem persisted, but at a >slower rate. >Any tips to correct this problem would be greatly appreciated. > >Becky Proctor >UNC-Chapel Hill > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Jan 10 16:00:52 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] CD21, B220, CD4 In-Reply-To: <000201c4f757$51e16d40$76d48a80@AMY> References: <000201c4f757$51e16d40$76d48a80@AMY> Message-ID: <6.0.0.22.1.20050110144809.01b516d8@gemini.msu.montana.edu> Liz, As discussed many times on Histonet, CD4 will NEVER work on FFPE! FYI, CD8 also will not work. If you have to run all these antibodies on the same sample, you will have to do frozen sections in order to accomadate the CD4. FS are air dried overnight at RT over dessicant. To improve morphology, use 75% acetone/25% absolute ethanol (no substitute on alcohol here) for 5 min at RT next day. Do NOT air dry again, go directly from fixative to buffer and then stain. We actually use biotinylated primaries more than purified, then come back with Strepavidin-HRP, then DAKO AEC+ or Chris van der Loos AEC - his formulation is excellent inhouse chromogen. Using biotinylated primaries shortens a protocol! We buy all antibodies, monoclonals from BD PharminGen. SEROTEC is another supplier. These are rat antimouse monoclonals. At 01:59 PM 1/10/2005, you wrote: >I need to run these antibodies on mouse tissue. Can anyone recommend me >a good source and if they might possibly work in FFPE tissue or do I >need to use frozens. > >Thanks > >Liz > >Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC >Premier Laboratory, LLC >P.O. Box 18592 >Boulder, Colorado 80308 >Office: (303) 735-5001 >Fax: (303) 735-3540 >liz@premierlab.com >www.premierlab.com > >Ship to Address: >Premier Laboratory >University of Colorado >MCDB, Room A3B40 >Boulder, Colorado 80309 > >_______________________________________________ >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 northma <@t> ohsu.edu Mon Jan 10 16:50:44 2005 From: northma <@t> ohsu.edu (Mary North) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Sections floating off of slides Message-ID: Another step to help adhesion of the section is rehydration of the block face before sectioning. If the section appears to be chalky-white on the slide it probably will not adhere well. Take care not to overrehydrate, especially brain, as it will swell and get mushy. If you have a warming plate, it also helps after draining excess water from under the section, to let it dry flat at 42 degrees C a few hours or overnight. The sections become translucent if they are properly dried. Mary North, HT, HTL(ASCP) Neuromuscular Laboratory OHSU Portland, OR >>> Gayle Callis 01/10/05 1:46 PM >>> Several things could be happening 1. No adhesive in either the waterbath - some add a few granules of gelatin as waterbath heats up or not using Plus Charge (silane or poly l lysine coated) slides to keep sections adhered to the slide surface. If you use Plus charge slides, the water should be distilled and not tap water as minerals in tap water in certain parts of the country will attract to the Plus charge and negate section adherance. 2. If you use ammonia water to blue the hematoxylin, it may be chewing the protein sections off the slides aka alkaline protein hydrolysis which makes sections release. Change to Scotts tap water substitute to blue you sections. This problem can occure even after oven drying of sections. If you use Ammonia water, it should be very dilute, just enough to blue the section, change it daily since it is a cheap solution. Sections can release with ammonia water bluing even IF you use slide adhesives. 3. Hopefully, your brain sections are very flat to perfect contact with the slide. Sharp blades are a must. At 01:31 PM 1/10/2005, you wrote: >Hello All, >I have encountered a problem H&E staining my paraffin embedded 8um mouse brain >sections. The sections are floating right off some of the slides but some of >the samples are normal. >The sectioning was done between mid-November 2004 through last week. The ones >sectioned more recently were embedded simultaneously and seem to be fine. >I tried baking the slides at 55 C for an hour, the problem persisted, but at a >slower rate. >Any tips to correct this problem would be greatly appreciated. > >Becky Proctor >UNC-Chapel Hill > >_______________________________________________ >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 bwhitaker <@t> brownpathology.com Mon Jan 10 17:33:29 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] CALIBRATING SCALES In-Reply-To: <41E31AE8.7020301@umdnj.edu> Message-ID: <000201c4f76c$ca709140$3601a8c0@brownpathology.net> I would see if someone in your area has a set of traceable weights that you could use to validate your weights. You could take your weights to their institution, weigh yours and theirs and record the difference, etc. I think that with sufficient documentation to back up your weights, the ones you have could be validated for use. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Geoff McAuliffe Sent: Monday, January 10, 2005 6:17 PM To: Winters, Bert Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] CALIBRATING SCALES Hi Brian: I must admit that I was skeptical until I looked in the Fischer catalogue, a complete set of weights lists for $980. That said, what is the matter with your "old" weights? Maybe if you need great accuracy to 4 decimal places you might need some very precise weights, but otherwise I think not. What sort of reagent needs that degree of precision? When we had Mettler come out and calibrate out balances the technician always had his own set of weights. I would talk to the manufacturer, the vendor and whoever is providing service. Geoff Winters, Bert wrote: >We have a new balance that needs to be calibrated by our biomed. >department before we can put it in service. The weights that we having >using for years are no longer the correct standard for calibrating the >scale and biomed tells us we need to order special weight that will >exceed $1,000. Does anyone know of a service that will come out and >calibrate the scale. That seems to be a more cost efficient way of >taking care of this problem. Any advice would be greatly appreciated. > > > Bert Winters, Northwest community hospital > > BWINTERS@NCH.ORG > > >******************* PLEASE NOTE ******************* >This E-Mail/telefax message and any documents accompanying this >transmission may contain information that is privileged, confidential, and/or exempt from disclosure under applicable law and is intended solely for the addressee(s) named above. If you are not the intended addressee/recipient, you are hereby notified that any use of, disclosure, copying, distribution, or reliance on the contents of this E-Mail/telefax information is strictly prohibited and may result in legal action against you. Please reply to the sender advising of the error in transmission and immediately delete/destroy the message and any accompanying documents. Thank you. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From weeks <@t> uoneuro.uoregon.edu Mon Jan 10 20:50:43 2005 From: weeks <@t> uoneuro.uoregon.edu (Janis C. Weeks) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] reversible tissue adhesion to slides Message-ID: <200501110252.j0B2qTUA019259@uoneuro.uoregon.edu> Dear colleagues, We are looking for a method to very firmly attach unfixed larval Drosophila ventral ganglia to a glass slide so we can slice off one surface of the ganglion (to remove neurons that we don't want; it's a long story; e-mail me if you want to know), to be followed by *releasing the attached tissue* from the slide and preparing primary neuronal cultures. The tissue needs to be kept alive and happy under saline. Polylysine doesn't hold tight enough for the slicing step (which we do with a fine blade). Possibilities that have come to mind are using more adhesive slides (e.g., coated with aminosilane?), using some sort of double-stick tape, or a releasable glue (e.g., something like vetbond but that can be made to release?). Obviously part of the difficulty is to release the tissue without destroying it. Does anyone have a suggestion? thanks! Janis Weeks --=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- Janis C. Weeks, Professor of Biology weeks@uoneuro.uoregon.edu http://www.neuro.uoregon.edu/faculty/weeks.html Institute of Neuroscience 1254 University of Oregon Eugene, OR 97403-1254 voice (541)346-4517, FAX (541)346-4548 --=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- From histology.bc <@t> shaw.ca Mon Jan 10 21:45:46 2005 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Sections floating off of slides In-Reply-To: <20050110153148.fv63ghmn0ggsssos@webmail3.isis.unc.edu> References: <20050110153148.fv63ghmn0ggsssos@webmail3.isis.unc.edu> Message-ID: <41E34BEA.8000006@shaw.ca> Hi Becky, Keeping brain sections on the slide has always been a problem. Over the years, I have tried plain slides, coated slides, textured slides, etc. Sometimes they seem to work, sometimes they fail, there does not seem to be a guaranteed method. One adhesive that was very successful is methyl cellulose. A small amount sprinkled on the suface of the water bath and allowed to dissolve was probably the best brain adhesive I have tried. Methyl cellulose is available in most paint/decorating stores as an adhesive for wallpaper. In low concentrations, it does not seem to react with most stains and dyes, so the background stays clear. I have no idea how it reacts with immunohistochemical techniques, I have not used this stuff since IHC became routine. Give it a try, it might work for you too. Pick up the sections onto the slides, allow them to drain and dry them for an hour or two at 60 degrees. Paul Bradbury Kamloops, BC Canada From jkiernan <@t> uwo.ca Mon Jan 10 22:59:59 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] reversible tissue adhesion to slides References: <200501110252.j0B2qTUA019259@uoneuro.uoregon.edu> Message-ID: <41E35D4F.637A8267@uwo.ca> How about this? (No, I've not tried it!) 1. Put a very thin tiny spot of cyanoacrylate (crazy glue) on the slide, followed by the ganglion. Expose to water vapour for a few minutes (to prevent drying, and to ensure that all the cyanoacrylate polymerizes - the reaction is catalyzed by water, metal ions etc). 2. Slice off the cells you don't want. 3. Shave the ganglion off the slide by sliding an inclined razor blade along the glass. 4. Put the removed ganglion in culture medium or whatever comes next. I'd guess that the neurons contacted by cyanoacrylate monomer would die. They might remain stuck to the slide. All the other neuronal cell bodies and the central core of neuropil might be OK. Cyanoacrylate glue is used for sticking unfixed specimens to the chuck of a vibrating microtome, and it does hold the specimens very strongly while they are being sectioned immersed in liquid (water, saline etc according to requirements). Cyanoacrylate adhesives are used as tissue adhesives (instead of stitches); they work well in wet places like the mouth, where sticky tape would be no good. If the polymerized glue is harmless in vivo it might also be harmless if some bits of it end up in your culture. Finally, if you can slice off the surface of the ganglion that you don't want, why can't you slice off and collect the part that you do want to keep? That way, no glue would enter the culture. John Kiernan Anatomy Dept, U.W.O., London, Canada. ________________________________ "Janis C. Weeks" wrote: > > Dear colleagues, > > We are looking for a method to very firmly attach unfixed larval > Drosophila ventral ganglia to a glass slide so we can slice off one > surface of the ganglion (to remove neurons that we don't want; it's a > long story; e-mail me if you want to know), to be followed by > *releasing the attached tissue* from the slide and preparing primary > neuronal cultures. The tissue needs to be kept alive and happy > under saline. Polylysine doesn't hold tight enough for the slicing > step (which we do with a fine blade). Possibilities that have come > to mind are using more adhesive slides (e.g., coated with > aminosilane?), using some sort of double-stick tape, or a > releasable glue (e.g., something like vetbond but that can be made > to release?). Obviously part of the difficulty is to release the tissue > without destroying it. Does anyone have a suggestion? thanks! > > Janis Weeks > > --=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- > Janis C. Weeks, Professor of Biology > weeks@uoneuro.uoregon.edu > http://www.neuro.uoregon.edu/faculty/weeks.html > Institute of Neuroscience > 1254 University of Oregon > Eugene, OR 97403-1254 > voice (541)346-4517, FAX (541)346-4548 > --=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From t.j.a.vaneijl <@t> pharm.uu.nl Tue Jan 11 04:20:50 2005 From: t.j.a.vaneijl <@t> pharm.uu.nl (Sven van Eijl) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Re: Mouse lung fixation In-Reply-To: <20050110183557.37153401A@mail.uu.nl> References: <20050110183557.37153401A@mail.uu.nl> Message-ID: <6.1.2.0.0.20050111102640.037d5050@solismail.uu.nl> At 07:35 10-01-05, you wrote: >In a message dated 1/9/2005 5:44:15 AM US Mountain Standard Time, >Andrew.MacDuff@ed.ac.uk writes: >am just starting a research project and need to fix mouse lungs for embedding >in parafin so that I can use them for histology and immunohistochemistry. >Could you please tell me the best way to fix the lungs? So far I have tried >removing the lungs and then instilling 10% neutral buffered formalin into the >trachea with a syringe. This seems to work but I'm not sure how >reproducible the >alveolar architecture will be. Also do people exsanguinate the mouse before >fixing the tissues? (we've been using cervical dislocation and then just >removing >the lungs) >Andrew, Hi Andrew, Are you going to do any morphometry on the alveoli ? in that case it is important that the lungs are always fixed at a set pressure. We take the lungs out of the body and attach them by the trachea to a set-up in which we keep the column of fixative which is pressurizing the lungs at 25 cm (through the communicating vessels principle). This way you can ensure that each lung is inflated to the same extent. We use Carnoy's, which acts very rapidly, but also seems to leak through the tissue quite a bit once it is fixed, probably because it is chloroform-based. Personally, I rather not use cervical dislocation, afraid it might damage lung structures, but I'm not sure that's really an issue. Sven -- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.300 / Virus Database: 265.6.10 - Release Date: 10-01-05 From hodges420 <@t> msn.com Tue Jan 11 05:20:01 2005 From: hodges420 <@t> msn.com (MARY T HODGES) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] controls needed Message-ID: Good morning all, I hope you all had and wonderful holiday. I am in need of two controls for IHC. One a CMV control and the other a mesotheleoma (sorry I misspelled it )control. If any on would like to trade I have cocci and pneumo control coming out our ears Thanks Tere Hodges St Mary's Hospital From Kristopher.Kalleberg <@t> unilever.com Tue Jan 11 07:39:12 2005 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Azur B Message-ID: HAs anyone used Azur(e) B for a melanin stain. I read the other day that you can use Azur B to stain malanin a green-blue, but everything else I read only says it is used for plant tissue. If anyone has used it for melanin, how does it work and what would be the best counterstain. Thanks. Kris L Kalleberg Unilever R&D 45 River Rd. Edgewater, NJ 07020 201 840 2472 From peoshel <@t> wisc.edu Tue Jan 11 08:32:46 2005 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] reversible tissue adhesion to slides In-Reply-To: <200501110252.j0B2qTUA019259@uoneuro.uoregon.edu> References: <200501110252.j0B2qTUA019259@uoneuro.uoregon.edu> Message-ID: Janis, How about attaching the part you *don't* want? That way the dissection will also release the desired tissue. Use a minute (no! smaller! still smaller ... ) drop of superglue to hold the tissue to the slide -- the same glue used to glue fresh tissue to the mounting plate of a vibrating microtome. Phil >Dear colleagues, > >We are looking for a method to very firmly attach unfixed larval >Drosophila ventral ganglia to a glass slide so we can slice off one >surface of the ganglion (to remove neurons that we don't want; it's a >long story; e-mail me if you want to know), to be followed by >*releasing the attached tissue* from the slide and preparing primary >neuronal cultures. The tissue needs to be kept alive and happy >under saline. Polylysine doesn't hold tight enough for the slicing >step (which we do with a fine blade). Possibilities that have come >to mind are using more adhesive slides (e.g., coated with >aminosilane?), using some sort of double-stick tape, or a >releasable glue (e.g., something like vetbond but that can be made >to release?). Obviously part of the difficulty is to release the tissue >without destroying it. Does anyone have a suggestion? thanks! > >Janis Weeks > >--=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- >Janis C. Weeks, Professor of Biology >weeks@uoneuro.uoregon.edu >http://www.neuro.uoregon.edu/faculty/weeks.html >Institute of Neuroscience >1254 University of Oregon >Eugene, OR 97403-1254 >voice (541)346-4517, FAX (541)346-4548 >--=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From antje.marcantonio <@t> pharma.novartis.com Tue Jan 11 08:49:30 2005 From: antje.marcantonio <@t> pharma.novartis.com (antje.marcantonio@pharma.novartis.com) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] CD68 in monkey frozen tissue - the solution Message-ID: For those of you who asked me to share the results: Beautiful staining with clone Ki-M6 from Serotec, dilution 1:50, over night incubation Cheers, Antje Antje Marcantonio Novartis Pharma AG BU Transplantation Research Basel, Switzerland Tel. +41 61 324 6730 antje.marcantonio@pharma.novartis.com From vazquezr <@t> ohsu.edu Tue Jan 11 09:05:02 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] Sections floating off of slides Message-ID: Paul, Would this work with frozen nasal bone? Robyn OHSU >>> Paul Bradbury 1/10/2005 7:45:46 PM >>> Hi Becky, Keeping brain sections on the slide has always been a problem. Over the years, I have tried plain slides, coated slides, textured slides, etc. Sometimes they seem to work, sometimes they fail, there does not seem to be a guaranteed method. One adhesive that was very successful is methyl cellulose. A small amount sprinkled on the suface of the water bath and allowed to dissolve was probably the best brain adhesive I have tried. Methyl cellulose is available in most paint/decorating stores as an adhesive for wallpaper. In low concentrations, it does not seem to react with most stains and dyes, so the background stays clear. I have no idea how it reacts with immunohistochemical techniques, I have not used this stuff since IHC became routine. Give it a try, it might work for you too. Pick up the sections onto the slides, allow them to drain and dry them for an hour or two at 60 degrees. Paul Bradbury Kamloops, BC Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Lawrence.Brett <@t> luht.scot.nhs.uk Tue Jan 11 09:03:52 2005 From: Lawrence.Brett <@t> luht.scot.nhs.uk (Brett, Lawrence) Date: Fri Sep 16 15:24:28 2005 Subject: FW: [Histonet] Azur B Message-ID: <857344E7D8F90240935A288998A89D811E775E@wgh-ex1.luht.scot.nhs.uk> Kristipher Yes we use Azure B regularly as a counterstain for S100 or other immunocytochemistry for melanoma. Use a simple 1% aqueous solution of Azure B for 5 minutes after the immuno, it turns the melanin green, making a nice contrast to the brown immunostain. We don't usually counterstain it, you may have to stain with haematoxylin first. Lawrence Brett Royal Infirmary Edinburgh Scotland, UK -----Original Message----- From: Kristopher Kalleberg [mailto:Kristopher.Kalleberg@unilever.com] Sent: 11 January 2005 13:39 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Azur B HAs anyone used Azur(e) B for a melanin stain. I read the other day that you can use Azur B to stain malanin a green-blue, but everything else I read only says it is used for plant tissue. If anyone has used it for melanin, how does it work and what would be the best counterstain. Thanks. Kris L Kalleberg Unilever R&D 45 River Rd. Edgewater, NJ 07020 201 840 2472 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** 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 kbroomal <@t> NEMOURS.ORG Tue Jan 11 09:18:31 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:24:28 2005 Subject: [Histonet] CD21, B220, CD4 Message-ID: <9BCAC308B27CAD4196D8AC9ADF037AAA110A78F9@wlmmsx01.nemours.org> Gayle, We've been using CD4 & CD8 on FFPE tissues with not much problem. They are from Neomarkers (www.labvision.com). CD4 Ab-8 http://www.labvision.com/ab.cfm?first=AntiBody&second=1528 CD8 Ab-1 http://www.labvision.com/ab.cfm?first=AntiBody&second=457 Kristen Broomall, HT (ASCP) -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Monday, January 10, 2005 5:01 PM To: Elizabeth Chlipala; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] CD21, B220, CD4 Liz, As discussed many times on Histonet, CD4 will NEVER work on FFPE! FYI, CD8 also will not work. If you have to run all these antibodies on the same sample, you will have to do frozen sections in order to accomadate the CD4. FS are air dried overnight at RT over dessicant. To improve morphology, use 75% acetone/25% absolute ethanol (no substitute on alcohol here) for 5 min at RT next day. Do NOT air dry again, go directly from fixative to buffer and then stain. We actually use biotinylated primaries more than purified, then come back with Strepavidin-HRP, then DAKO AEC+ or Chris van der Loos AEC - his formulation is excellent inhouse chromogen. Using biotinylated primaries shortens a protocol! We buy all antibodies, monoclonals from BD PharminGen. SEROTEC is another supplier. These are rat antimouse monoclonals. At 01:59 PM 1/10/2005, you wrote: >I need to run these antibodies on mouse tissue. Can anyone recommend me >a good source and if they might possibly work in FFPE tissue or do I >need to use frozens. > >Thanks > >Liz > >Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC >Premier Laboratory, LLC >P.O. Box 18592 >Boulder, Colorado 80308 >Office: (303) 735-5001 >Fax: (303) 735-3540 >liz@premierlab.com >www.premierlab.com > >Ship to Address: >Premier Laboratory >University of Colorado >MCDB, Room A3B40 >Boulder, Colorado 80309 > >_______________________________________________ >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 Dorothy.L.Webb <@t> HealthPartners.Com Tue Jan 11 09:51:50 2005 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] testicular biopsies Message-ID: What is the preferred fixative that is used for testicular biopsies, especially when dealing with fertility studies. Does anyone have a preference in using Bouin's, Zenker's, or gluteraldehyde and is there a special procedure to be followed when using any of those fixatives? Thanks ahead of time for your help in this area of concern!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From cornettl <@t> hotmail.com Tue Jan 11 09:57:36 2005 From: cornettl <@t> hotmail.com (Lorraine Cornett) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] AFB Message-ID: Does anyone have extra AFB control blocks that they could spare? We are cutting our last speck out of the block and haven't had any positive cases to go back to for control tissue. Thanks Lorraine Cornett, HT (ASCP) Highlands Pathology Blue Ridge Division, Kingsport, TN 423 224-5793 fax 423 224-5349 _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! hthttp://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From pruegg <@t> ihctech.net Tue Jan 11 10:20:33 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] reversible tissue adhesion to slides In-Reply-To: Message-ID: <200501111620.j0BGKX2p011572@chip.viawest.net> I release super glued tissue using a single edge razor blade. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Philip Oshel Sent: Tuesday, January 11, 2005 7:33 AM To: Histonet@Pathology.swmed.edu Subject: Re: [Histonet] reversible tissue adhesion to slides Janis, How about attaching the part you *don't* want? That way the dissection will also release the desired tissue. Use a minute (no! smaller! still smaller ... ) drop of superglue to hold the tissue to the slide -- the same glue used to glue fresh tissue to the mounting plate of a vibrating microtome. Phil >Dear colleagues, > >We are looking for a method to very firmly attach unfixed larval >Drosophila ventral ganglia to a glass slide so we can slice off one >surface of the ganglion (to remove neurons that we don't want; it's a >long story; e-mail me if you want to know), to be followed by >*releasing the attached tissue* from the slide and preparing primary >neuronal cultures. The tissue needs to be kept alive and happy under >saline. Polylysine doesn't hold tight enough for the slicing step >(which we do with a fine blade). Possibilities that have come to mind >are using more adhesive slides (e.g., coated with aminosilane?), using >some sort of double-stick tape, or a releasable glue (e.g., something >like vetbond but that can be made to release?). Obviously part of the >difficulty is to release the tissue >without destroying it. Does anyone have a suggestion? thanks! > >Janis Weeks > >--=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- Janis C. Weeks, >Professor of Biology weeks@uoneuro.uoregon.edu >http://www.neuro.uoregon.edu/faculty/weeks.html >Institute of Neuroscience >1254 University of Oregon >Eugene, OR 97403-1254 >voice (541)346-4517, FAX (541)346-4548 >--=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- --=[|]=-- > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Tue Jan 11 10:21:42 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Azur B In-Reply-To: Message-ID: <200501111621.j0BGLf2p011905@chip.viawest.net> I use Azure B as a proteoglycan stain, it will stain different tissue components depending on the ph. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristopher Kalleberg Sent: Tuesday, January 11, 2005 6:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Azur B HAs anyone used Azur(e) B for a melanin stain. I read the other day that you can use Azur B to stain malanin a green-blue, but everything else I read only says it is used for plant tissue. If anyone has used it for melanin, how does it work and what would be the best counterstain. Thanks. Kris L Kalleberg Unilever R&D 45 River Rd. Edgewater, NJ 07020 201 840 2472 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Tue Jan 11 10:37:04 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Azur B Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0F352FC@UTHEVS3.mail.uthouston.edu> As far as I remember the sequence is methylene blue, azure B, azure A, azure 4, azure C, Bernthen's methylene violet and thionin. This is a progression and, in general the dyes having greater metachromatic properties the closer that they are to thionin and pH dependent. Barry ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Patsy Ruegg Sent: Tue 1/11/2005 10:21 AM To: 'Kristopher Kalleberg'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Azur B I use Azure B as a proteoglycan stain, it will stain different tissue components depending on the ph. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristopher Kalleberg Sent: Tuesday, January 11, 2005 6:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Azur B HAs anyone used Azur(e) B for a melanin stain. I read the other day that you can use Azur B to stain malanin a green-blue, but everything else I read only says it is used for plant tissue. If anyone has used it for melanin, how does it work and what would be the best counterstain. Thanks. Kris L Kalleberg Unilever R&D 45 River Rd. Edgewater, NJ 07020 201 840 2472 _______________________________________________ Histonet mailing list Histonet@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 MAUGER <@t> email.chop.edu Tue Jan 11 11:22:00 2005 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] CD4, cd8 Message-ID: To All, We have used CD4 and CD8 from Novocastra with great success on FFPE. Also CD4 from Zymed works on FFPE- all need EDTA retreival. Jo >>> "Kristen Broomall" 01/11/05 10:18 AM >>> Gayle, We've been using CD4 & CD8 on FFPE tissues with not much problem. They are from Neomarkers (www.labvision.com). CD4 Ab-8 http://www.labvision.com/ab.cfm?first=AntiBody&second=1528 CD8 Ab-1 http://www.labvision.com/ab.cfm?first=AntiBody&second=457 Kristen Broomall, HT (ASCP) -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Monday, January 10, 2005 5:01 PM To: Elizabeth Chlipala; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] CD21, B220, CD4 Liz, As discussed many times on Histonet, CD4 will NEVER work on FFPE! FYI, CD8 also will not work. If you have to run all these antibodies on the same sample, you will have to do frozen sections in order to accomadate the CD4. FS are air dried overnight at RT over dessicant. To improve morphology, use 75% acetone/25% absolute ethanol (no substitute on alcohol here) for 5 min at RT next day. Do NOT air dry again, go directly from fixative to buffer and then stain. We actually use biotinylated primaries more than purified, then come back with Strepavidin-HRP, then DAKO AEC+ or Chris van der Loos AEC - his formulation is excellent inhouse chromogen. Using biotinylated primaries shortens a protocol! We buy all antibodies, monoclonals from BD PharminGen. SEROTEC is another supplier. These are rat antimouse monoclonals. At 01:59 PM 1/10/2005, you wrote: >I need to run these antibodies on mouse tissue. Can anyone recommend me >a good source and if they might possibly work in FFPE tissue or do I >need to use frozens. > >Thanks > >Liz > >Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC >Premier Laboratory, LLC >P.O. Box 18592 >Boulder, Colorado 80308 >Office: (303) 735-5001 >Fax: (303) 735-3540 >liz@premierlab.com >www.premierlab.com > >Ship to Address: >Premier Laboratory >University of Colorado >MCDB, Room A3B40 >Boulder, Colorado 80309 > >_______________________________________________ >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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Thomas.Crowell <@t> biogenidec.com Tue Jan 11 11:41:40 2005 From: Thomas.Crowell <@t> biogenidec.com (Thomas Crowell) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Humanized mouse monoclonals In-Reply-To: Message-ID: Dear Histonetters, I was wondering if anyone could provide me with a basic protocol using a preformed complex of a humanized mouse monoclonal (human Fc ) with a biotinylated anti-human IgG to detect antigens in human cryostat sections - essentially, human on human IHC staining? I have several antibodies with different total protein concentrations. Thanks Tom Crowell BiogenIDEC Cambridge, MA From jbrod <@t> tvmdl.tamu.edu Tue Jan 11 11:48:28 2005 From: jbrod <@t> tvmdl.tamu.edu (Jordan Brod) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] To Anyone who has the DLM Certification: Message-ID: I am seeking advice from anyone who has this certification. I believe that it may benefit my career in the future, if not immediately. My problem is: I don't have a lot of time to dedicate to studying. >From your experience(s), how difficult is the exam. Would you assume that a laboratory supervisor of five years, who deals with most of the criteria tasks daily could pass the test with little preparation? What study book/guide is proven to be the best? I want to be able to maximize my prep. time with the best materials. Any help would be appreciated. Thanks. Jordan From lhadley <@t> iupui.edu Tue Jan 11 12:04:29 2005 From: lhadley <@t> iupui.edu (Baldridge, Lee Ann) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] CD4, cd8 Message-ID: Hi Everyone, If you check the original post you'll find we're talking about mouse tissue. We too have great success with cd4 & cd8 but in ffpe Human tissue not mouse. Good Luck, Lee Ann Baldridge IUSM Indpls,IN. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joanne Mauger Sent: Tuesday, January 11, 2005 12:22 PM To: Histonet@lists.utsouthwestern.edu; gcallis@montana.edu; kbroomal@NEMOURS.ORG; liz@premierlab.com Subject: RE: [Histonet] CD4, cd8 To All, We have used CD4 and CD8 from Novocastra with great success on FFPE. Also CD4 from Zymed works on FFPE- all need EDTA retreival. Jo >>> "Kristen Broomall" 01/11/05 10:18 AM >>> Gayle, We've been using CD4 & CD8 on FFPE tissues with not much problem. They are from Neomarkers (www.labvision.com). CD4 Ab-8 http://www.labvision.com/ab.cfm?first=AntiBody&second=1528 CD8 Ab-1 http://www.labvision.com/ab.cfm?first=AntiBody&second=457 Kristen Broomall, HT (ASCP) -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Monday, January 10, 2005 5:01 PM To: Elizabeth Chlipala; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] CD21, B220, CD4 Liz, As discussed many times on Histonet, CD4 will NEVER work on FFPE! FYI, CD8 also will not work. If you have to run all these antibodies on the same sample, you will have to do frozen sections in order to accomadate the CD4. FS are air dried overnight at RT over dessicant. To improve morphology, use 75% acetone/25% absolute ethanol (no substitute on alcohol here) for 5 min at RT next day. Do NOT air dry again, go directly from fixative to buffer and then stain. We actually use biotinylated primaries more than purified, then come back with Strepavidin-HRP, then DAKO AEC+ or Chris van der Loos AEC - his formulation is excellent inhouse chromogen. Using biotinylated primaries shortens a protocol! We buy all antibodies, monoclonals from BD PharminGen. SEROTEC is another supplier. These are rat antimouse monoclonals. At 01:59 PM 1/10/2005, you wrote: >I need to run these antibodies on mouse tissue. Can anyone recommend me >a good source and if they might possibly work in FFPE tissue or do I >need to use frozens. > >Thanks > >Liz > >Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC >Premier Laboratory, LLC >P.O. Box 18592 >Boulder, Colorado 80308 >Office: (303) 735-5001 >Fax: (303) 735-3540 >liz@premierlab.com >www.premierlab.com > >Ship to Address: >Premier Laboratory >University of Colorado >MCDB, Room A3B40 >Boulder, Colorado 80309 > >_______________________________________________ >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 _______________________________________________ Histonet mailing list Histonet@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 Thomas.Crowell <@t> biogenidec.com Tue Jan 11 12:08:41 2005 From: Thomas.Crowell <@t> biogenidec.com (Thomas Crowell) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Re: Histonet Digest, Vol 14, Issue 11 In-Reply-To: Message-ID: Dear Histonetters, I'd like to hear your opinions on antibodies that are particularly useful to asses the overall immunoreactivity of human epithelial tumors in various tissue microarrays from commercial sources. I'm interested in using antibodies that would have a broad range of reactivity in these tumors, and are fixation sensitive. Thanks in advance Tom Crowell BiogenIDEC Cambridge, MA From ohenry <@t> dfw.net Tue Jan 11 12:46:46 2005 From: ohenry <@t> dfw.net (Susan Owens) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] HT needed in Texas Message-ID: <000901c4f80d$e98b4ba0$9ddd3040@Nationwide.net> Well guys and gals, we need a full time HT here in Arlington Texas (located between Dallas and Ft.Worth) Cutting,embedding,gross room FS, special stains. If you might be interested contact: Joyce Martin 817-548-6233 or email joycemartin@AMHTX.com Susan Susan Owens-TX ohenry@dfw.net voice: 817-261-7938 fax: 817-548-9876 "A bad day at the dog show is better then a good day at work!" From kbroomal <@t> NEMOURS.ORG Tue Jan 11 13:03:16 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] CD4, cd8 Message-ID: <9BCAC308B27CAD4196D8AC9ADF037AAA110A7900@wlmmsx01.nemours.org> I was just commenting on this statement. "As discussed many times on Histonet, CD4 will NEVER work on FFPE! FYI, CD8 also will not work." -----Original Message----- From: Baldridge, Lee Ann [mailto:lhadley@iupui.edu] Sent: Tuesday, January 11, 2005 1:04 PM To: Joanne Mauger; Histonet@lists.utsouthwestern.edu; gcallis@montana.edu; kbroomal@NEMOURS.ORG; liz@premierlab.com Subject: RE: [Histonet] CD4, cd8 Hi Everyone, If you check the original post you'll find we're talking about mouse tissue. We too have great success with cd4 & cd8 but in ffpe Human tissue not mouse. Good Luck, Lee Ann Baldridge IUSM Indpls,IN. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joanne Mauger Sent: Tuesday, January 11, 2005 12:22 PM To: Histonet@lists.utsouthwestern.edu; gcallis@montana.edu; kbroomal@NEMOURS.ORG; liz@premierlab.com Subject: RE: [Histonet] CD4, cd8 To All, We have used CD4 and CD8 from Novocastra with great success on FFPE. Also CD4 from Zymed works on FFPE- all need EDTA retreival. Jo >>> "Kristen Broomall" 01/11/05 10:18 AM >>> Gayle, We've been using CD4 & CD8 on FFPE tissues with not much problem. They are from Neomarkers (www.labvision.com). CD4 Ab-8 http://www.labvision.com/ab.cfm?first=AntiBody&second=1528 CD8 Ab-1 http://www.labvision.com/ab.cfm?first=AntiBody&second=457 Kristen Broomall, HT (ASCP) -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Monday, January 10, 2005 5:01 PM To: Elizabeth Chlipala; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] CD21, B220, CD4 Liz, As discussed many times on Histonet, CD4 will NEVER work on FFPE! FYI, CD8 also will not work. If you have to run all these antibodies on the same sample, you will have to do frozen sections in order to accomadate the CD4. FS are air dried overnight at RT over dessicant. To improve morphology, use 75% acetone/25% absolute ethanol (no substitute on alcohol here) for 5 min at RT next day. Do NOT air dry again, go directly from fixative to buffer and then stain. We actually use biotinylated primaries more than purified, then come back with Strepavidin-HRP, then DAKO AEC+ or Chris van der Loos AEC - his formulation is excellent inhouse chromogen. Using biotinylated primaries shortens a protocol! We buy all antibodies, monoclonals from BD PharminGen. SEROTEC is another supplier. These are rat antimouse monoclonals. At 01:59 PM 1/10/2005, you wrote: >I need to run these antibodies on mouse tissue. Can anyone recommend me >a good source and if they might possibly work in FFPE tissue or do I >need to use frozens. > >Thanks > >Liz > >Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC >Premier Laboratory, LLC >P.O. Box 18592 >Boulder, Colorado 80308 >Office: (303) 735-5001 >Fax: (303) 735-3540 >liz@premierlab.com >www.premierlab.com > >Ship to Address: >Premier Laboratory >University of Colorado >MCDB, Room A3B40 >Boulder, Colorado 80309 > >_______________________________________________ >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 _______________________________________________ Histonet mailing list Histonet@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 mgr <@t> seattleskincancer.com Tue Jan 11 13:31:30 2005 From: mgr <@t> seattleskincancer.com (Manager) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] job in Seattle Message-ID: <20050111193324.8AF086B0A0@smtp4.pacifier.net> Hi, We have a part time job opening in Seattle cutting frozen sections for Mohs surgery. I am willing to train. Beth Uri, HT (ASCP) Seattle Skin Cancer Center Seattle, WA mgr@seattleskincancer.com From MTitford <@t> aol.com Tue Jan 11 13:38:52 2005 From: MTitford <@t> aol.com (MTitford@aol.com) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Repeat in-situ diagnosis question Message-ID: <056C0D1C.2E6026D6.00762DB1@aol.com> I did not get any feed back to an earlier question so I would like to try it again: Are any histology labs using in-situ hybridization to diagnose tumors? using commercially available kits? (Where the tumor is caused by a single DNA abnormality?) Is this going to be the next big thing to follow immunohistochemistry? Thank you Mike Titford USA Pathology Mobile AL USA From WWmn916 <@t> aol.com Tue Jan 11 13:39:43 2005 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] MS access and histology Message-ID: <3A0500F7.25F1304C.001A2AA1@aol.com> Hello histonetters, Has anyone had any experience using microsoft access and its applications with histology? Deb King, HT(ASCP) Path Logic Sacramento, CA From Matthew_Frank <@t> URMC.Rochester.edu Tue Jan 11 14:01:13 2005 From: Matthew_Frank <@t> URMC.Rochester.edu (Frank, Matthew) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] MS access and histology Message-ID: I use access frequently. It is just a database program. Nothing specific for histology about it. It isn't intended for large data sets like a Lab Information System LIS or networked entry where a central data server is needed. You could use it to set up supplier files or just about anything on a small-med scale. However, it isn't the easiest program to learn if your not computer savy. Does that help at all? Matthew C. Frank MBA,HTL Biological Stain Commission University of Rochester Pathology Department Box 626 601 Elmwood Avenue Rochester, New York 14642-0001 (585) 275-2751 -----Original Message----- From: WWmn916@aol.com [mailto:WWmn916@aol.com] Sent: Tuesday, January 11, 2005 2:40 PM To: histonet@pathology.swmed.edu Subject: [Histonet] MS access and histology Hello histonetters, Has anyone had any experience using microsoft access and its applications with histology? Deb King, HT(ASCP) Path Logic Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Jan 11 14:09:36 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:29 2005 Subject: Question was responded to about antibodies for MOUSE tissue [Histonet] CD21, B220, CD4 In-Reply-To: References: Message-ID: <6.0.0.22.1.20050111130508.01b7d458@gemini.msu.montana.edu> To set the record straight, Liz was asking about doing CD4, and the other markers on MOUSE tissue using FFPE. The answer is - no, CD4 does not work on murine tissue FFPE, only frozen sections. Somehow human tissue crept into this messaging which we know works very well with CD4 and CD8. Doggone mouse is a problem however. At 08:29 AM 1/11/2005, you wrote: >CD4 and CD8 work for us. > >CD4 (Novacastra) antigen retreived in citrate buffer, or trilogy work >fine. CD8(Dako) antigen retreive in citrate buffer. > >Doug Geddes BSc, MLT >London Health Sciences Centre >London Ontario, Canada > > >>> Gayle Callis 01/10/05 05:00PM >>> >Liz, > >As discussed many times on Histonet, CD4 will NEVER work on FFPE! FYI, >CD8 >also will not work. > >If you have to run all these antibodies on the same sample, you will >have >to do frozen sections in order to accomadate the CD4. FS are air dried > >overnight at RT over dessicant. To improve morphology, use 75% >acetone/25% absolute ethanol (no substitute on alcohol here) for 5 min >at >RT next day. Do NOT air dry again, go directly from fixative to buffer >and >then stain. We actually use biotinylated primaries more than purified, > >then come back with Strepavidin-HRP, then DAKO AEC+ or Chris van der >Loos >AEC - his formulation is excellent inhouse chromogen. Using >biotinylated >primaries shortens a protocol! > >We buy all antibodies, monoclonals from BD PharminGen. SEROTEC is >another >supplier. These are rat antimouse monoclonals. > >At 01:59 PM 1/10/2005, you wrote: > >I need to run these antibodies on mouse tissue. Can anyone recommend >me > >a good source and if they might possibly work in FFPE tissue or do I > >need to use frozens. > > > >Thanks > > > >Liz > > > >Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > >Premier Laboratory, LLC > >P.O. Box 18592 > >Boulder, Colorado 80308 > >Office: (303) 735-5001 > >Fax: (303) 735-3540 > >liz@premierlab.com > >www.premierlab.com > > > >Ship to Address: > >Premier Laboratory > >University of Colorado > >MCDB, Room A3B40 > >Boulder, Colorado 80309 > > > >_______________________________________________ > >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 information is directed in confidence solely to the person named above >and may contain confidential and/or privileged material. This information >may not otherwise be distributed, copied or disclosed. If you have >received this e-mail in error, please notify the sender immediately via a >return e-mail and destroy original message. Thank you for your >cooperation. From mward <@t> wfubmc.edu Tue Jan 11 14:27:55 2005 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] MLH1 and MSH2 antibodies Message-ID: <61135F0455D33347B5AAE209B903A3040BBFFBC8@EXCHVS2.medctr.ad.wfubmc.edu> I recently had a pathologist inquire about adding these antibodies to our test menu. Is anyone doing these currently? The only vendor he suggested was BD Biosciences. Any information would be appreciated. Thanks! Martha Ward, MT (ASCP) QIHC From dbpiontek <@t> clinomicsbio.com Tue Jan 11 14:34:47 2005 From: dbpiontek <@t> clinomicsbio.com (Denise Bland-Piontek) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] FW: Job Opportunity in New York Message-ID: <145F33B84793CF488578F86355B32E691704DB@mail.clinomicsbio.com> Lead Histotechnologist Clinomics Biosciences, Inc. is seeking a lead histotechnologist for operation of our Histopathology Laboratory. The candidate will be expected to oversee IHC (many antibody characterizations), TMA (paraffin and frozen) and daily duties for a staff of five. HT or HTL certification required with five plus years of experience. CAP accredited laboratory, maintenance for inspections. Knowledge of in situ hybridization, immunofluoresence and TUNEL a plus. Laser Capture Microdissection is performed in the laboratory. CoolScope is utilized for client interaction and digital library. This is a great opportunity to step outside the box and be a direct participant in a research role. Extremely competitive salary. Clinomics Biosciences is located in Watervliet, NY on Rensselaer Polytechnic Institute campus just outside of Albany, NY. Please contact Denise Bland-Piontek, dbpiontek@clinomicsbio.com (617) 938-1250 for application or questions. Please visit our website www.clinomicsbio.com . Denise Bland-Piontek, HTL(ASCP)CTBS(AATB) Clinomics Biosciences, Inc. Histopathology Manager (617) 938-1250 dbpiontek@clinomicsbio.com From Jackie.O'Connor <@t> abbott.com Tue Jan 11 14:43:10 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Adam-9 Antibody Message-ID: These requests get weirder and weirder - Anyone have a line on an Adam-9 antibody for IHC? I find several for WB - I'd rather find one already proven for IHC. Thank you thank you thank you. Jackie O' From ohenry <@t> dfw.net Tue Jan 11 15:07:31 2005 From: ohenry <@t> dfw.net (Susan Owens) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] HT needed in Texas Message-ID: <003501c4f821$9e6d7800$7bdd3040@Nationwide.net> To answer a couple of questions. Yes, this is for Arlington Memorial Hospital. AMH is located in North Arlington, about 5 minutes from the baseball stadium(Texas Rangers) and if things go as voted for last Nov. the new home of the Dallas Cowboys will be 2 minutes pass the baseball stadium...In other words, a 10 minute or less drive, due east of the hospital. Arlington is a real nice city to live and work in. Susan ---------------------------------------------------------------------------- ---- Well guys and gals, we need a full time HT here at Arlington Memorial Hospital in Arlington Texas (located between Dallas and Ft.Worth). Cutting,embedding,gross room FS, special stains. If you might be interested contact: Joyce Martin 817-548-6233 or email joycemartin@AMHTX.com Susan Susan Owens-TX ohenry@dfw.net voice: 817-261-7938 fax: 817-548-9876 "A bad day at the dog show is better then a good day at work!" From WWmn916 <@t> aol.com Tue Jan 11 15:19:15 2005 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] MS access and histology Message-ID: <713CB459.0E2A9D06.001A2AA1@aol.com> Hello again I'm thinking of setting up a system for not only supplies and inventory....but to use access for logging request (levels, recuts, ipox request, special stains, etc.) I hoping to get rid of binders of manually logged paper tasks, print monthly tables, be able to extract information as I build my data base for use in statistics. There is a two day Access training seminar coming up soon. I'm doing ebook instructionals beforehand. I found myself wondering how extensively Access is being used in histology for logging request, special stains and ipox. Any suggestions are very welcomed. Deb King Sacramento, CA From histo20 <@t> hotmail.com Tue Jan 11 15:37:04 2005 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] CAP question for Immuno Message-ID: The CAP question concerning the temperature of the tissue and slide processing for immuno - AP22400 - How is everyone else responding to this question? Incorporated into our procedures is the temperature of the slide drying oven, but the inspectors felt that that information was insufficient and the tissue temps also need to be incorporated into the procedures. Histology does have a daily recording of paraffin bath temperatures and the temperature range - is that what they want? Thanks a bunch Paula Wilder St. Joseph Medical Center Towson, MD 21204 From victor <@t> pathology.washington.edu Tue Jan 11 15:45:51 2005 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] HT needed in Texas In-Reply-To: <003501c4f821$9e6d7800$7bdd3040@Nationwide.net> References: <003501c4f821$9e6d7800$7bdd3040@Nationwide.net> Message-ID: <41E4490F.5000206@pathology.washington.edu> And the signing bonus is season passes? Susan Owens wrote: >To answer a couple of questions. Yes, this >is for Arlington Memorial Hospital. AMH is located in North Arlington, about >5 minutes from the baseball stadium(Texas Rangers) and if things go as voted >for last Nov. the new home of the Dallas Cowboys will be 2 minutes pass the >baseball stadium...In other words, a 10 minute or less drive, due east of >the hospital. >Arlington is a real nice city to live and work in. > > >Susan > > >---------------------------------------------------------------------------- >---- > >Well guys and gals, we need a full time HT here at Arlington Memorial >Hospital > in Arlington Texas (located between Dallas and Ft.Worth). > >Cutting,embedding,gross room FS, special stains. > >If you might be interested contact: > >Joyce Martin 817-548-6233 >or email >joycemartin@AMHTX.com > > >Susan >Susan Owens-TX >ohenry@dfw.net >voice: 817-261-7938 >fax: 817-548-9876 > >"A bad day at the dog show is better then a good day at work!" > > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. From colesy1222 <@t> yahoo.com Tue Jan 11 16:06:27 2005 From: colesy1222 <@t> yahoo.com (Nicole Puglisi) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] microwave processing Message-ID: <20050111220627.18643.qmail@web30602.mail.mud.yahoo.com> I am wondering how many of you out there have Milestone RHS-1 and RHS-2 microwave processors? How are they working for you? Another question, how often do you feel you would need to order replacement parts (histomodules, pyrex containers, racks, lids, etc.)? I've looked into the price and it is pretty steep. Any replies are welcome and helpful! Thanks! --------------------------------- Do you Yahoo!? The all-new My Yahoo! – Get yours free! From Jackie.O'Connor <@t> abbott.com Tue Jan 11 16:06:41 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] MS access and histology Message-ID: Access was a little too much for some of the PhD's to comprehend here - so we went to a very simple program, FileMakerPro. I use it to log virtually everything - Accession numbers, workload, IHC requests - you name it. I designed the database to fit my needs, so of course I love it. My only gripe is that you don't have to hit a save key when you enter information - so if you make a mistake and accidentally hit the wrong key or enter data in the wrong file, you'd never know it - well, until it was found later. Besides that - it's great. It has a lock out function, too, so that other people cant enter data - we have it on our common drive so the info is accessible to everyone, but only a select few can enter selected data. Jackie WWmn916@aol.com Sent by: histonet-bounces@lists.utsouthwestern.edu 01/11/2005 03:19 PM To: histonet@pathology.swmed.edu cc: Subject: [Histonet] MS access and histology Hello again I'm thinking of setting up a system for not only supplies and inventory....but to use access for logging request (levels, recuts, ipox request, special stains, etc.) I hoping to get rid of binders of manually logged paper tasks, print monthly tables, be able to extract information as I build my data base for use in statistics. There is a two day Access training seminar coming up soon. I'm doing ebook instructionals beforehand. I found myself wondering how extensively Access is being used in histology for logging request, special stains and ipox. Any suggestions are very welcomed. Deb King Sacramento, CA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DWeil <@t> ciit.org Tue Jan 11 16:12:38 2005 From: DWeil <@t> ciit.org (David Weil) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] RE: Mouse Lung Fixation Message-ID: <12816CB59E68184F9F5F28063E68B04701D35CD6@xsrvr.ciit.org> Dear Andrew Investigators here have had very good results inflating mouse lungs for both histology and immunohistochemistry endpoints using this procedure. It preserves the architecture and causes minimal damage to alveolar architecture. A column of 10% neutral buffered formalin is suspended 20 cm above work surface using clamp stand. An extension tube is connected from the formalin column to a blunt 20 gauge needle. The tubing is primed before inflation so no air passed into the lungs The mouse is anesthetized with an injectable anesthetic (pentobarbital). Open the abdominal cavity and then exanguinate by cutting the abdominal aorta. It is useful reveal the trachea at this time . The thoracic cavity is cut open carefully to avoid cutting the lungs or any vessels. Cut a small incisions in the trachea making sure no to cut it in half completely. Insert the 20 gauge needle into the trachea and secure it with a piece of suture. Let the formalin flow into the lungs until fully inflated. Tie off the lungs with a piece of suture so no formalin escapes. Remove the trachea, heart, thymus and lungs and place in a cup of formalin for 48 hours. Switch the lungs to 70% ethanol until ready for gross trim and processing. I do not know how cervical dislocation will affect this procedure, but we have had very good results with this method and it gives the investigators reproducible results. If you have any further questions please contact me. Good luck with the project. David S Weil BS, HTL CIIT Centers For Health Research 6 Davis Dr. RTP, NC 27709-2137 (919) 558-1265 dweil@ciit.org From LuckG <@t> empirehealth.org Tue Jan 11 16:18:49 2005 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Trying to contact Michael Lafriniere Message-ID: Hello, Does anyone have an email address for Michael Lafriniere? Thanks Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org From amosbrooks <@t> earthlink.net Tue Jan 11 16:32:43 2005 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] CD4 & CD8 In-Reply-To: <200501110854.1cOpi854W3NZFpK0@mx-a065b01.pas.sa.earthlink. net> References: <200501110854.1cOpi854W3NZFpK0@mx-a065b01.pas.sa.earthlink.net> Message-ID: <6.2.0.14.0.20050111172214.01d7ab30@pop.earthlink.net> Liz & Gayle, Actually we've had really good luck with both CD4 & CD8. The trick with CD4 is doing a peroxidase block before retrieval or not at all. If you do it after retrieval it will really diminish the reactivity. Use high pH epitope retrieval (I'm using DAKO for this in a steamer for 20 min). You should also use a non-biotin secondary antibody as the retrieval really brings out the endogenous biotin. Try Impress from Vector it's great! For a primary antibody we also use Neomarkers for both. Try around 1:10 - 1:20 for a dilution. Best of luck, Amos Brooks At 11:54 AM 1/11/2005, you wrote: >Message: 8 >Date: Mon, 10 Jan 2005 15:00:52 -0700 >From: Gayle Callis >Subject: Re: [Histonet] CD21, B220, CD4 >To: "Elizabeth Chlipala" , > Histonet@lists.utsouthwestern.edu >Message-ID: > <6.0.0.22.1.20050110144809.01b516d8@gemini.msu.montana.edu> >Content-Type: text/plain; charset="us-ascii"; format=flowed > >Liz, > >As discussed many times on Histonet, CD4 will NEVER work on FFPE! FYI, CD8 >also will not work. > >If you have to run all these antibodies on the same sample, you will have >to do frozen sections in order to accomadate the CD4. FS are air dried >overnight at RT over dessicant. To improve morphology, use 75% >acetone/25% absolute ethanol (no substitute on alcohol here) for 5 min at >RT next day. Do NOT air dry again, go directly from fixative to buffer and >then stain. We actually use biotinylated primaries more than purified, >then come back with Strepavidin-HRP, then DAKO AEC+ or Chris van der Loos >AEC - his formulation is excellent inhouse chromogen. Using biotinylated >primaries shortens a protocol! > >We buy all antibodies, monoclonals from BD PharminGen. SEROTEC is another >supplier. These are rat antimouse monoclonals. From bwhitaker <@t> brownpathology.com Tue Jan 11 16:44:27 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] MS access and histology In-Reply-To: Message-ID: <000001c4f82f$1b2027a0$3601a8c0@brownpathology.net> Maybe Tim Morken will speak up here..... He has used Access extensively, and very successfully. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie M O'Connor Sent: Tuesday, January 11, 2005 4:07 PM To: WWmn916@aol.com Cc: histonet-bounces@lists.utsouthwestern.edu; histonet@pathology.swmed.edu Subject: Re: [Histonet] MS access and histology Access was a little too much for some of the PhD's to comprehend here - so we went to a very simple program, FileMakerPro. I use it to log virtually everything - Accession numbers, workload, IHC requests - you name it. I designed the database to fit my needs, so of course I love it. My only gripe is that you don't have to hit a save key when you enter information - so if you make a mistake and accidentally hit the wrong key or enter data in the wrong file, you'd never know it - well, until it was found later. Besides that - it's great. It has a lock out function, too, so that other people cant enter data - we have it on our common drive so the info is accessible to everyone, but only a select few can enter selected data. Jackie WWmn916@aol.com Sent by: histonet-bounces@lists.utsouthwestern.edu 01/11/2005 03:19 PM To: histonet@pathology.swmed.edu cc: Subject: [Histonet] MS access and histology Hello again I'm thinking of setting up a system for not only supplies and inventory....but to use access for logging request (levels, recuts, ipox request, special stains, etc.) I hoping to get rid of binders of manually logged paper tasks, print monthly tables, be able to extract information as I build my data base for use in statistics. There is a two day Access training seminar coming up soon. I'm doing ebook instructionals beforehand. I found myself wondering how extensively Access is being used in histology for logging request, special stains and ipox. Any suggestions are very welcomed. Deb King Sacramento, CA _______________________________________________ Histonet mailing list Histonet@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 tpmorken <@t> labvision.com Tue Jan 11 17:30:51 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] MS access and histology Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA257F86D@usca0082k08.labvision.apogent.com> Thanks for the plug Bonnie, and now about my NSH Access for Histology workshop -- hopefully we'll have it again in 2005! Anyway, Access is a powerful program, and small databases can be put together very quickly, but it does have a steep learning curve for advanced use. And you need a little visual basic programming to get the full benefit. FilemakerPro is "easier" in the sense that the macro language is plain english and so easier to comprehend. I've done extensive databases in both and think both are good. There are quite a few commercial LIS applications out there built on Access. One caveat about building databases for lab use: Make sure someone besides the original designer knows how it works, because if that person leaves, the application may quickly become useless as no one can fix it or modify it! Tim Morken Lab Vision - Neomarkers www.labvision.com Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonnie Whitaker Sent: Tuesday, January 11, 2005 2:44 PM To: 'Jackie M O'Connor'; WWmn916@aol.com Cc: histonet-bounces@lists.utsouthwestern.edu; histonet@pathology.swmed.edu Subject: RE: [Histonet] MS access and histology Maybe Tim Morken will speak up here..... He has used Access extensively, and very successfully. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie M O'Connor Sent: Tuesday, January 11, 2005 4:07 PM To: WWmn916@aol.com Cc: histonet-bounces@lists.utsouthwestern.edu; histonet@pathology.swmed.edu Subject: Re: [Histonet] MS access and histology Access was a little too much for some of the PhD's to comprehend here - so we went to a very simple program, FileMakerPro. I use it to log virtually everything - Accession numbers, workload, IHC requests - you name it. I designed the database to fit my needs, so of course I love it. My only gripe is that you don't have to hit a save key when you enter information - so if you make a mistake and accidentally hit the wrong key or enter data in the wrong file, you'd never know it - well, until it was found later. Besides that - it's great. It has a lock out function, too, so that other people cant enter data - we have it on our common drive so the info is accessible to everyone, but only a select few can enter selected data. Jackie WWmn916@aol.com Sent by: histonet-bounces@lists.utsouthwestern.edu 01/11/2005 03:19 PM To: histonet@pathology.swmed.edu cc: Subject: [Histonet] MS access and histology Hello again I'm thinking of setting up a system for not only supplies and inventory....but to use access for logging request (levels, recuts, ipox request, special stains, etc.) I hoping to get rid of binders of manually logged paper tasks, print monthly tables, be able to extract information as I build my data base for use in statistics. There is a two day Access training seminar coming up soon. I'm doing ebook instructionals beforehand. I found myself wondering how extensively Access is being used in histology for logging request, special stains and ipox. Any suggestions are very welcomed. Deb King Sacramento, CA _______________________________________________ Histonet mailing list Histonet@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 barnhart717 <@t> comcast.net Tue Jan 11 17:31:09 2005 From: barnhart717 <@t> comcast.net (Becky Barnhart) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] MS access and histology References: <000001c4f82f$1b2027a0$3601a8c0@brownpathology.net> Message-ID: <000701c4f835$a25b3ce0$2b285344@Tuckey> I built an access database to save my statistics. We have Meditech as an LIS and I run monthly reports and enter the data into my access database. This way I can more easily see the specific information I want. Also not everyone has access to the reports in Meditech and to get some of the information there are calculations and it is much easier just to put a date range in and let the computer do all the calculations. Our pathologist loves it. I am at home right now so I can't look in to see all the reports I have but some are: 1.) Workload broken out into surgical blocks and slides, cytology blocks and slides, autopsy blocks and slides, recuts, special stains, conventional pap smears and liquid base paps (I think that is it). 2.) The amount of specimen we receive in just from those outside our regular service area. 3.) Cytotech for her annual and 6 month reviews 4.) Surgical service committee that shows how many cases (surgical, Non gyn, Gyn), how many frozen section, FNA and consults and if they all agree or disagree. 5.) Our turnaround times for each specimen type 6.) Amount we charge out each month All of the reports prompt for a date range so I can get the amount of data I need. If I can be of any help let me know. You may contact me either at home or work (rbarnhart@summithealth.org) is probably better so I can look at my database. Becky Barnhart ----- Original Message ----- From: "Bonnie Whitaker" To: "'Jackie M O'Connor'" ; Cc: ; Sent: Tuesday, January 11, 2005 5:44 PM Subject: RE: [Histonet] MS access and histology > Maybe Tim Morken will speak up here..... He has used Access extensively, > and > very successfully. > > Bonnie Whitaker > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie M > O'Connor > Sent: Tuesday, January 11, 2005 4:07 PM > To: WWmn916@aol.com > Cc: histonet-bounces@lists.utsouthwestern.edu; > histonet@pathology.swmed.edu > Subject: Re: [Histonet] MS access and histology > > > Access was a little too much for some of the PhD's to comprehend here - > so we went to a very simple program, FileMakerPro. I use it to log > virtually everything - Accession numbers, workload, IHC requests - you > name it. I designed the database to fit my needs, so of course I love it. > My only gripe is that you don't have to hit a save key when you enter > information - so if you make a mistake and accidentally hit the wrong key > or enter data in the wrong file, you'd never know it - well, until it was > found later. Besides that - it's great. It has a lock out function, too, > so that other people cant enter data - we have it on our common drive so > the info is accessible to everyone, but only a select few can enter > selected data. > Jackie > > > > > WWmn916@aol.com > Sent by: histonet-bounces@lists.utsouthwestern.edu > 01/11/2005 03:19 PM > > > To: histonet@pathology.swmed.edu > cc: > Subject: [Histonet] MS access and histology > > > Hello again > I'm thinking of setting up a system for not only supplies and > inventory....but to use access for logging request (levels, recuts, ipox > request, special stains, etc.) I hoping to get rid of binders of manually > logged paper tasks, print monthly tables, be able to extract information > as I build my data base for use in statistics. > > There is a two day Access training seminar coming up soon. I'm doing > ebook instructionals beforehand. I found myself wondering how extensively > Access is being used in histology for logging request, special stains and > ipox. > > Any suggestions are very welcomed. > > Deb King > Sacramento, CA > > _______________________________________________ > Histonet mailing list > Histonet@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 jnocito <@t> satx.rr.com Tue Jan 11 18:28:25 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] CAP question for Immuno References: Message-ID: <009001c4f83d$a16d39c0$cf2df318@yourxhtr8hvc4p> Paula, just having survived my CAP inspection yesterday, my inspector also wanted to see the temperature logs for heat retrieval also. When we have to perform antibodies off the Ventana XT, we use the Decloaking Chamber from Biocare Medical. They have temperature strips that turn black after a certain temperature, something like the tape that Micro uses to autoclave. I can send you a copy of what we use. When I was using a steamer, I used to put a thermometer in the retrieval solution and log the temperature on a log sheet. Hope this helps and congratulations for surviving another CAP inspection. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Paula Wilder" To: Sent: Tuesday, January 11, 2005 3:37 PM Subject: [Histonet] CAP question for Immuno > The CAP question concerning the temperature of the tissue and slide > processing for immuno - AP22400 - How is everyone else responding to this > question? Incorporated into our procedures is the temperature of the > slide drying oven, but the inspectors felt that that information was > insufficient and the tissue temps also need to be incorporated into the > procedures. Histology does have a daily recording of paraffin bath > temperatures and the temperature range - is that what they want? Thanks a > bunch > Paula Wilder > St. Joseph Medical Center > Towson, MD 21204 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brian.dias <@t> mail.utexas.edu Tue Jan 11 19:31:32 2005 From: brian.dias <@t> mail.utexas.edu (Brian Dias) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Sections being chewed up. Message-ID: <5.2.0.9.2.20050111192452.00bdee78@mail.utexas.edu> hi everyone, i'm trying to get my immunohistochemistry on slides to work for several antigens and have been successful in doing so in the past whilst preserving tissue morphology on the slides at the same time. of late, i've been getting tissue that looks chewed up, pock-marked (really badly)after i get thru' the ICC (this happens to both my fresh frozen as well as perfused section slides, more with the fresh frozen). the antigen staining still exists BUT is extremely hazy due to the tissue morphology. any input is welcome. thanks a ton regards b ps just thought this might be of interest to the list users: in the past i had issues with my 30um sections falling off slides whilst performing my ICC. that problem was solved by using Superfrost Plus Gold slides from Erie Scientific (thanks barbara!!!) ____________________________________________________________________________ Brian George Dias Graduate Student (Crews lab) Office Phone #: 512-475-6738 The University of Texas at Austin E-mail: brian.dias@mail.utexas.edu Institute for Neuroscience Website: https://webspace.utexas.edu/bgd85/b.htm Patterson 56 1 University Station Stop C0930 Austin, TX 78712 ____________________________________________________________________________ From pengbw <@t> sjtu.edu.cn Tue Jan 11 20:07:37 2005 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Re: Mouse Lung Fixation Message-ID: <20050112020737.18F7C111D9CB@sjtu.edu.cn> Dear David, I\'m wondering if I can use OCT instead of formaloin to inflate mouse lung for frozen sections as decribed in your protocol. Thanks a lot! ----- Original Message ----- From: "David Weil" To: Cc: Subject: [Histonet] RE: Mouse Lung Fixation Dear Andrew Investigators here have had very good results inflating mouse lungs for both histology and immunohistochemistry endpoints using this procedure. It preserves the architecture and causes minimal damage to alveolar architecture. A column of 10% neutral buffered formalin is suspended 20 cm above work surface using clamp stand. An extension tube is connected from the formalin column to a blunt 20 gauge needle. The tubing is primed before inflation so no air passed into the lungs The mouse is anesthetized with an injectable anesthetic (pentobarbital). Open the abdominal cavity and then exanguinate by cutting the abdominal aorta. It is useful reveal the trachea at this time . The thoracic cavity is cut open carefully to avoid cutting the lungs or any vessels. Cut a small incisions in the trachea making sure no to cut it in half completely. Insert the 20 gauge needle into the trachea and secure it with a piece of suture. Let the formalin flow into the lungs until fully inflated. Tie off the lungs with a piece of suture so no formalin escapes. Remove the trachea, heart, thymus and lungs and place in a cup of formalin for 48 hours. Switch the lungs to 70% ethanol until ready for gross trim and processing. I do not know how cervical dislocation will affect this procedure, but we have had very good results with this method and it gives the investigators reproducible results. If you have any further questions please contact me. Good luck with the project. David S Weil BS, HTL CIIT Centers For Health Research 6 Davis Dr. RTP, NC 27709-2137 (919) 558-1265 dweil@ciit.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Baowei Peng 1954 Huashan Road Pharmacy School Shanghai Jiaotong University Shanghai, 200030 China Ph:86-21-62932108 E-mail:pengbw@sjtu.edu.cn From Krat18 <@t> aol.com Wed Jan 12 03:48:04 2005 From: Krat18 <@t> aol.com (Krat18@aol.com) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Static electricity problems Message-ID: <1f2.40fe53c.2f164c54@aol.com> Our lab has sectioning problems in the winter because of static electricity and low humidity. Can anyone tell me what is the ideal humidity for good quality sectioning in histology, and if the humidity is too low in the lab (ours runs between 32% and 38% in the winter), what is the best way to raise it without creating a draft or other sectioning difficulties? Thanks for your advice. karen_raterman@ssmhc.com Karen Raterman St. Mary's Health Center St. Louis, MO 63117 From Kemlo.Rogerson <@t> elht.nhs.uk Wed Jan 12 04:05:23 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Static electricity problems[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EE2F@bhrv-nt-11.bhrv.nwest.nhs.uk> Plants. Loads of plants, especially Monstera Deliciousa. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Krat18@aol.com [mailto:Krat18@aol.com] Sent: 12 January 2005 09:48 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Static electricity problems[Scanned] Our lab has sectioning problems in the winter because of static electricity and low humidity. Can anyone tell me what is the ideal humidity for good quality sectioning in histology, and if the humidity is too low in the lab (ours runs between 32% and 38% in the winter), what is the best way to raise it without creating a draft or other sectioning difficulties? Thanks for your advice. karen_raterman@ssmhc.com Karen Raterman St. Mary's Health Center St. Louis, MO 63117 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Wed Jan 12 04:27:18 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] RE: Humanized mouse monoclonals Message-ID: Dear Tom, For the detection of a "humanized" monoclonal antibody on human tissue one need a procedure similar to the ARKit: in vitro labeling of the primary antibody with a biotinylated goat anti-IgG (of primary species) Fab-fragment. Such a kit is available from Molecular Probes and called a Zenon-kit (http://www.probes.com/products/zenon/zenon.html). According to their website it is available for mouse, rabbit and human primary antibodies and the label is either biotin or a fluorochrome. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 NL-1105 AZ Amsterdam The Netherlands ----- Original Message ----- >From Thomas Crowell Date Tue, 11 Jan 2005 12:41:40 -0500 To histonet@lists.utsouthwestern.edu Subject [Histonet] Humanized mouse monoclonals Dear Histonetters, I was wondering if anyone could provide me with a basic protocol using a preformed complex of a humanized mouse monoclonal (human Fc ) with a biotinylated anti-human IgG to detect antigens in human cryostat sections - essentially, human on human IHC staining? I have several antibodies with different total protein concentrations. Thanks Tom Crowell BiogenIDEC Cambridge, MA From ajennings <@t> unmc.edu Wed Jan 12 08:29:13 2005 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Mouse Lung inflation-OCT In-Reply-To: <20050112020737.18F7C111D9CB@sjtu.edu.cn> Message-ID: OCT is almost impossible to use to inflate the lungs, it is very viscous, but I have used a 1:1 OCT and 5%sucrose solution to inflate adult mouse lungs and it adds tremendously to the morphology of the frozen lung tissue and makes it easier to section also. Immunohistochemistry and in situ procedures have been performed on these samples with no adverse effects. From pmarcum <@t> polysciences.com Wed Jan 12 08:38:01 2005 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Static electricity problems In-Reply-To: <1f2.40fe53c.2f164c54@aol.com> Message-ID: <001201c4f8b4$514a44c0$7f00a8c0@PMARCUM2K> Good Morning, Several things can help even if raising the humidity to 50% or higher is not possible. One of the first things I learned was to watch what I wear in the winter. Many of the polyester, acrylic or other man-made fabrics develop static electricity in the cold and it is good to touch something to discharge any built up static on you or your clothing by touching something metal often. If you see that telltale spark or feel the charge you are adding to the problem. I place a dryer sheet like Bounce on the back of my knife stage and if it gets really bad use it to wipe down the surfaces of the microtome and my hands. They are cheap and do help a little. I have tried using a humidifier near the microtome and is limited in how much help it can be as the heating system is taking it out as fast I can put it in. Don't have any alcohol or xylene near the cutting are if possible that brings it down more. Plants do help if you have sunlight and space. They won't solve the whole problem. (I only rubber sole shoes in the lab.) Hope something here helps. Best Regards, Pamela A Marcum Histology/Microscopy Product Development Manager Polysciences, Inc. 400 Valley Road Warrington, PA 18976 Telephone: 800-523-2575???? Ext. 167 215-343-6484???? Ext. 167 Fax:? 800-343-3291 ???????? 215-343-0214 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of > Krat18@aol.com > Sent: Wednesday, January 12, 2005 4:48 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Static electricity problems > > > Our lab has sectioning problems in the winter because of static > electricity > and low humidity. Can anyone tell me what is the ideal humidity > for good > quality sectioning in histology, and if the humidity is too low > in the lab > (ours runs between 32% and 38% in the winter), what is the best > way to raise it > without creating a draft or other sectioning difficulties? > > Thanks for your advice. > > karen_raterman@ssmhc.com > Karen Raterman > St. Mary's Health Center > St. Louis, MO 63117 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From SAllen <@t> exchange.hsc.mb.ca Wed Jan 12 08:55:40 2005 From: SAllen <@t> exchange.hsc.mb.ca (Sharon Allen) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Precipitate of Bielschowsky slides. Message-ID: <304B5A264EC9974E8121B0EA14A9C3936619D7@hsc01mx1.hsc.mb.ca> Hi, We have in the past made our own AAS slides, but due to cost we are buying commercial ones now. We have noticed that on our Bielschowsky's (silver staining) we are getting a precipitate. Has anyone else had this problem & have a solution? It is very distracting for the neuropathologist reading the slides. Thanks Sharon sallen@hsc.mb.ca This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. From Sue.Tyler <@t> noaa.gov Wed Jan 12 09:29:46 2005 From: Sue.Tyler <@t> noaa.gov (Sue Tyler) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] acid clean glassware Message-ID: <41E5426A.2080008@noaa.gov> I have searched the archives for info. on replacing potassium dichromate sulfuric acid glass cleaner. I came up with a few suggestions namely, nitric acid or a Clorox bleach wash. I was wondering what your current opinions are of these suggestions. Do you still like them and do they work as well as suggested? Is there a commercial cleaner that works just as well? -- Sue Tyler Biologist Cooperative Oxford Laboratory Center for Coastal Environmental Health & Biomolecular Research at Charleston ( CCHEBR) USDOC/NOAA/NOS/NCCOS 904 S. Morris St. Oxford, Maryland 21654-9724 410-226-5193 Fax: 410- 226-5925 Sue.Tyler@noaa.gov From JQB7 <@t> CDC.GOV Wed Jan 12 09:42:22 2005 From: JQB7 <@t> CDC.GOV (Bartlett, Jeanine) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] acid clean glassware Message-ID: I have always used 20% nitric with excellent results. 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 Sue Tyler Sent: Wednesday, January 12, 2005 10:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] acid clean glassware I have searched the archives for info. on replacing potassium dichromate sulfuric acid glass cleaner. I came up with a few suggestions namely, nitric acid or a Clorox bleach wash. I was wondering what your current opinions are of these suggestions. Do you still like them and do they work as well as suggested? Is there a commercial cleaner that works just as well? -- Sue Tyler Biologist Cooperative Oxford Laboratory Center for Coastal Environmental Health & Biomolecular Research at Charleston ( CCHEBR) USDOC/NOAA/NOS/NCCOS 904 S. Morris St. Oxford, Maryland 21654-9724 410-226-5193 Fax: 410- 226-5925 Sue.Tyler@noaa.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Jan 12 11:19:21 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Sorry to disagree! Re:Mouse Lung inflation-OCT In-Reply-To: References: <20050112020737.18F7C111D9CB@sjtu.edu.cn> Message-ID: <6.0.0.22.1.20050112101301.01b59df0@gemini.msu.montana.edu> Ann and others, Sorry to disagree, we do this routinely and never dilute the OCT. Introduction of OCT into lungs is via a v shaped cut on TOP of trachea. The trachea is never severed completely across the lumen or the trachea retracts into lung cavity. We use 2 mls OCT with a dulled 18 guage needle (we dull sharp point and edges with a fingernail file containing various grits sand paper!) inserted into the trachea gently towards the lung. After lung is OCT filled, gently remove lung from mouse, dissect heart away, and snap freeze. We experience no difficulty nor damage to lung morphology. We used to dilute the OCT with PBS 1:1, but found it was not necessary. At 07:29 AM 1/12/2005, you wrote: >OCT is almost impossible to use to inflate the lungs, it is very viscous, >but I have used a 1:1 OCT and 5%sucrose solution to inflate adult mouse >lungs and it adds tremendously to the morphology of the frozen lung tissue >and makes it easier to section also. Immunohistochemistry and in situ >procedures have been performed on these samples with no adverse effects. > > >_______________________________________________ >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 brian.dias <@t> mail.utexas.edu Wed Jan 12 11:42:55 2005 From: brian.dias <@t> mail.utexas.edu (Brian Dias) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] My ICC Protocol- Sections getting chewed Message-ID: <5.2.0.9.2.20050112113137.00bdfbd8@mail.utexas.edu> Protocol: Fresh frozen Animals decapped and brains frozen on dry ice. Stored at -80C. Sectioned (16um) sections on cryostat on to Superfrost Plus slides. Stored at -80C. On day of ICC, removed from -80C, thawed for 30 mins, ice-cold (-20C) acetone (10mins), air dried(30mins), 1XPBS-RT-5min(x2), 3%H2O2/1XPBS/0.3%TritonX-100 RT-30mins, 1XPBS-RT-5min(x2), Block (1XPBS/4%NGS/0.3%TritonX-100)-RT-1 hr, 1 Ab (1XPBS/2%NGS/0.3%TritonX-100)-RT-O/N, 1XPBS-RT-10min(x2), 2Ab (1XPBS/2%NGS/0.3%TritonX-100)-RT-2hrs, 1XPBS-RT-10min(x2), ABC-RT- 1hr, 1XPBS-RT-10min(x2), DAB-RT-10mins, 1XPBS-RT-10min(x2), air dry, dehydrate and coverslip Paraformaldehyde fixed Animals decapped and heads in 4%PFA-4C-5 hrs, Brains dissected, stored in 4%PFA-4C-3 hrs, Brains transferred to 20%sucrose/1XPBS-4C- approx. 12 hours, Sectioned (16um) sections on cryostat on to Superfrost Plus slides, Allowed to dry at RT O/N, Stored at -80C. On day of ICC, removed from -80C, thawed for 30 mins, 4%PFA-RT-10mins, 1XPBS-RT-5min(x2), 3%H2O2/1XPBS/0.3%TritonX-100 RT-30mins, 1XPBS-RT-5min(x2), 1% NaBorohydride in 1XPBS-RT-20mins, 1XPBS-RT-5min(x2), Block (1XPBS/4%NGS/0.3%TritonX-100)-RT-1 hr, 1 Ab (1XPBS/2%NGS/0.3%TritonX-100)-RT-O/N, 1XPBS-RT-10min(x2), 2Ab (1XPBS/2%NGS/0.3%TritonX-100)-RT-2hrs, 1XPBS-RT-10min(x2), ABC-RT- 1hr, 1XPBS-RT-10min(x2), DAB-RT-10mins, 1XPBS-RT-10min(x2), air dry, dehydrate and coverslip hi everyone, i'm trying to get my immunohistochemistry on slides to work for several antigens and have been successful in doing so in the past whilst preserving tissue morphology on the slides at the same time. of late, i've been getting tissue that looks chewed up, pock-marked (really badly)after i get thru' the ICC (this happens to both my fresh frozen as well as perfused section slides, more with the fresh frozen). the antigen staining still exists BUT is extremely hazy due to the tissue morphology. any input is welcome. thanks a ton regards b ps just thought this might be of interest to the list users: in the past i had issues with my 30um sections falling off slides whilst performing my ICC. that problem was solved by using Superfrost Plus Gold slides from Erie Scientific (thanks barbara!!!) ____________________________________________________________________________ Brian George Dias Graduate Student (Crews lab) Office Phone #: 512-475-6738 The University of Texas at Austin E-mail: brian.dias@mail.utexas.edu Institute for Neuroscience Website: https://webspace.utexas.edu/bgd85/b.htm Patterson 56 1 University Station Stop C0930 Austin, TX 78712 ____________________________________________________________________________ From MadaryJ <@t> MedImmune.com Wed Jan 12 13:13:43 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Boiling water near the microtome reduces static too Message-ID: <83899F0EC7671543B305FB5694024DE60A1B46D7@medimmune4.medimmune.com> From TMcNemar <@t> lmhealth.org Wed Jan 12 13:20:04 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Off-sitte storage of slides and blocks.. Message-ID: <6CD94D97ED7D924BA5C2B588FA952821396772@nt_exchange.lmhealth.org> Hello all, We are thinking of storing our blocks and slides off-site. We may contract with a company to store and maintain them, as well as dispose of them when the time comes. We would pay a monthly fee per cubic foot of space used and delivery charges when we need to get them back. Does anyone else do this? If so, how does it work for you? The problem is that we have no place to store them on-site. Our space was taken over by medical records and the slides and blocks were dispersed to any place they could find. They are stashed under and behind duct work, this corner and that. It's quite a mess. I'm thinking that off-site storage would be a good thing. They guarrentee delivery is as little as four hours. Does anyone charge to retrieve slides/blocks? I don't know if we could pass the retrieval/delivery charge on to the requestor..... Any thoughts appreciated. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 From funderwood <@t> mcohio.org Wed Jan 12 13:32:25 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:29 2005 Subject: [BULK] - [Histonet] Off-sitte storage of slides and blocks.. Message-ID: Hi Tom, Off site storage becomes a necessity for almost everyone at some point. I wouldn't put an unmonitored trust in anyone to watch over my blocks/slides. I would make certain that you also have access to the storage facility. It is important that you monitor temperature and humidity. I would recommend a good quality dehumidifier. It's also good to check on their handling of the specimens and make sure there are no rodents or insects having a buffet on all those tasty goodies. Fred >>> Tom McNemar 01/12/05 02:20PM >>> Hello all, We are thinking of storing our blocks and slides off-site. We may contract with a company to store and maintain them, as well as dispose of them when the time comes. We would pay a monthly fee per cubic foot of space used and delivery charges when we need to get them back. Does anyone else do this? If so, how does it work for you? The problem is that we have no place to store them on-site. Our space was taken over by medical records and the slides and blocks were dispersed to any place they could find. They are stashed under and behind duct work, this corner and that. It's quite a mess. I'm thinking that off-site storage would be a good thing. They guarrentee delivery is as little as four hours. Does anyone charge to retrieve slides/blocks? I don't know if we could pass the retrieval/delivery charge on to the requestor..... Any thoughts appreciated. 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 BlazekL <@t> childrensdayton.org Wed Jan 12 13:45:32 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Off-sitte storage of slides and blocks.. Message-ID: Tom, We found that sending things like old reports (the ones we use to have bound into books up until a couple of years ago) and all the extra equipment that just hangs around was better sent to off site storage than trusting them with blocks and slides. That left us with somewhat adequate space for our blocks and slides. 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 RossS <@t> BaylorHealth.edu Wed Jan 12 13:51:29 2005 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Off-sitte storage of slides and blocks.. Message-ID: <17A8DA5A05167C4EA20BFA750167736B0BB6C6@BHDAEXCH13.bhcs.pvt> We just had almost all of our storage moved off site as well. It is working ok. They charge to retrieve/deliver as well as a small amount to refile. The same company takes care of all off site storage for Baylor. When I was in Maryland we had off site storage at warehouse space the hospital maintained. If you wanted timely retrieval you had to drive and get it yourself. Also a previous supervisor had used the cardboard file boxes to store slides and over time multiple stacks had collapsed. We had to refile all of them ourselves. It took months of overtime to fix that problem, as well as hiring a college student on summer break to just work on refiling them into metal trays. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tom McNemar Sent: Wednesday, January 12, 2005 1:20 PM To: 'Histonet (histonet@pathology.swmed.edu)' Subject: [Histonet] Off-sitte storage of slides and blocks.. Hello all, We are thinking of storing our blocks and slides off-site. We may contract with a company to store and maintain them, as well as dispose of them when the time comes. We would pay a monthly fee per cubic foot of space used and delivery charges when we need to get them back. Does anyone else do this? If so, how does it work for you? The problem is that we have no place to store them on-site. Our space was taken over by medical records and the slides and blocks were dispersed to any place they could find. They are stashed under and behind duct work, this corner and that. It's quite a mess. I'm thinking that off-site storage would be a good thing. They guarrentee delivery is as little as four hours. Does anyone charge to retrieve slides/blocks? I don't know if we could pass the retrieval/delivery charge on to the requestor..... Any thoughts appreciated. 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 This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From POWELL_SA <@t> Mercer.edu Wed Jan 12 14:03:07 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Off-site storage of slides and blocks.. In-Reply-To: Message-ID: A caution to all who store off site. Once upon a time there was a hospital that stored slides, blocks and reports off site. One day "someone?" discovered patient reports, a lot of them, in the landfill where garbage was dumped, and the hospital found this out from the media. Be careful who you trust with your archives, make them sign the appropriate disclosure documents, make them accountable. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Linda Blazek Sent: Wednesday, January 12, 2005 2:46 PM To: TMcNemar@lmhealth.org; histonet@pathology.swmed.edu Subject: Re: [Histonet] Off-sitte storage of slides and blocks.. Tom, We found that sending things like old reports (the ones we use to have bound into books up until a couple of years ago) and all the extra equipment that just hangs around was better sent to off site storage than trusting them with blocks and slides. That left us with somewhat adequate space for our blocks and slides. 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpjones <@t> srhs-pa.org Wed Jan 12 14:15:53 2005 From: lpjones <@t> srhs-pa.org (Jones, Laura) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Mark Lewis at Shandon - HELP!! Message-ID: <8E7AD740937B954F947F0DB4467EFEE056E941@mail.srhs-pa.org> Hi Mark! If you are still at Shandon, could you please give us a call at 724-983-3914 ext.4875. We need your expertise regarding the Hypercenter XP and Glyofixx. Thanks! From DOOLEEO <@t> shands.ufl.edu Wed Jan 12 15:07:04 2005 From: DOOLEEO <@t> shands.ufl.edu (Elaine Dooley) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] bone marrow decal for insitu & immuno Message-ID: Dear Histonetters, We have been using Decal Stat (from Decal Corporation) on our bone marrow biopsies. It is HCL , EDTA and water. The sections stain well for most immunohistochemistry. But we would like to insitu hybridization for EBER and Kappa and Lambda. We have these tests working well on lymph nodes but they do not work on our bone marrow biopsies. I tested some bone marrow from an outside hospital and did get good Kappa and Lambda insitu staining. They use a formic acid decal mixture. (1000 cc 88%formic acid, 900cc water, 100cc 37% formaldehyde place bone marrow in decal 2 hours). So I processed and decalcified some bone marrow in this way and the insitu hybridization worked well. But the morphology on H&E is not as good as when we processed with stat decal. We have some Formical from Decal Corporation I could try. Does anyone use this to decal there bone marrow biopsies? If so how long? Or is there another formic acid based decal anyone would recommend that does not effect morphology upon H&E staining negatively? Elaine Dooley 352-265-0111 ext 72117 From katherine-walters <@t> uiowa.edu Wed Jan 12 16:53:24 2005 From: katherine-walters <@t> uiowa.edu (Walters, Katherine S) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] stain for tight junctions Message-ID: Hi all, I have kind of an interesting problem. We want to look at tight junctions in cells. We need a small molecule stain that will not be taken up by the cells. We want the stain to get between the cells and be stopped by the tight junctions. We do not want to use EM and we do not want to use fluorescence. Has anybody done this, or have an idea about how to do this? Thank you, Kathy Walters Central Microscopy Research Facility University of Iowa From robynm <@t> scripps.edu Wed Jan 12 18:24:37 2005 From: robynm <@t> scripps.edu (robynm) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] alpha/beta TCR antibody for IHC on human tissue Message-ID: <41E5DE7C@neo> Hi all, Does anybody know of a good biotin-conjugated anti-human alpha/beta TCR antibody I can order that works well for IHC on paraffin embedded tissue sections? Most of the companies that I've checked have only tested for flow cytometry and don't have information on IHC. Thanks for your help, Robyn From weneng2004 <@t> yahoo.com Wed Jan 12 18:51:55 2005 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] endothelial cell marker other than PECAM Message-ID: <20050113005155.14745.qmail@web53403.mail.yahoo.com> Hello, I am frustrated with CD31 IHC. Has anybody here successfully used other antibody? My tissue is FFPE mouse tissue. Thanks, Wen --------------------------------- Do you Yahoo!? The all-new My Yahoo! – What will yours do? From mike.kirby <@t> nhls.ac.za Thu Jan 13 04:37:10 2005 From: mike.kirby <@t> nhls.ac.za (Mike Kirby) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Monoethanolamine as a formalin neutralizer Message-ID: Hi Guys. Has anyone had any experience in using monoethanolamine as a formalin neutralizer? The reason I ask is that I came across an article in this regard, which stated that the chemical (Used in the automotive industry to clean parts) was found to be an effective neutralizer of formaldehyde, produced no precipitates, was cheap, and the resultant product was a nontoxic imine alcohol. We are familiar with the method of using ammonia as a neutralizer, and are aware that there are many commercial products in the same line, (Unfortunately, not available to us in the wilds of Africa), and we wondering whether monoethanolamine would be a viable alternative. Mike Kirby Johannesburg South Africa. ********************************************************************************** This disclaimer will apply to, but is not limited to a local employee, joint venture personnel, contractors, temporary personnel, outsourced service staff, client personnel working on cooperative engagements, alliance partners, Satellite offices or any other type of user as authorised by the National Health Laboratory Service (NHLS). Email transmission cannot be guaranteed to be secure or error free, as information may be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. The NHLS cannot ensure that the integrity of this communication is maintained nor that it is free from any error, viruses, interception or interference. Neither NHLS nor the sender accepts liability for any errors or omissions in the contents of this message which arise as a result of email transmission. If verification is required, please request a hard copy version. In no event will the NHLS or the sender be liable to anyone for any indirect, special, consequential or direct damages arising from this email or use thereof. No employee is authorised to conclude a binding agreement on behalf of the NHLS by email without the express written confirmation of the Executive Committee of the NHLS. Nothing contained in this email shall be construed as a legally binding agreement or an offer to contract. The use or contents of this email is intended for the NHLS business. If it is used for another purpose, the views and opinions expressed are those of the sender and no liability will attach to the NHLS. *********************************************************************************** From mhorne <@t> upei.ca Thu Jan 13 07:09:43 2005 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] endothelial cell marker other than PECAM In-Reply-To: <20050113005155.14745.qmail@web53403.mail.yahoo.com> Message-ID: <41E648E7.28381.233F3E@localhost> Hi , I haven't started yet , but am about to try von Willebrand's factor ( Factor VIII ). I'll be doing it on rat, first in FFPE to see if it works, and then on osmium fixed Epon embedded. Wish me luck! I chose Factor 8 because of a paper by Ulger et al , Anat Histol Embryol. 2002 Feb:31(1):31-5 where he compares various rat endothelial cell markers : Factor 8 , RECA-1, PECAM-1 , ICAM- 1, OX-43 and ZO-1. I am buying my anti body from Biocare Medical and their tech support suggests trying a pepsin treatment as they use it on the FFPE tissue at 37 C for 5 min. Hope this helps , don't hesitate to email me if you have any more questions, 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 DWeil <@t> ciit.org Thu Jan 13 08:19:10 2005 From: DWeil <@t> ciit.org (David Weil) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] RE: Histonet Digest, Vol 14, Issue 14 Message-ID: <12816CB59E68184F9F5F28063E68B04701D35E1F@xsrvr.ciit.org> Dear Baowei I have had only one project that involved inflating the lungs with OCT and it did not produce good results. However, the protocol that was being used called for several lung lavages with PBS before the lungs were inflated. The lavages leave fluid behind and it seemed to impair the inflation with OCT. The investigator did not continue with the procedure. We filled up the lung in a manner similar to that describe by Gayle. I am glad to hear that other folks have had success inflating lungs with OCT. Our attempts to use OCT for lung inflation just did not work for this particular project. David S Weil BS, HTL CIIT Centers For Health Research 6 Davis Dr. RTP, NC 27709-2137 (919) 558-1265 dweil@ciit.org From mcauliff <@t> umdnj.edu Thu Jan 13 12:21:46 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] stain for tight junctions In-Reply-To: References: Message-ID: <41E6BC3A.9000105@umdnj.edu> Hi Kathy: You can put peroxidase in the vascular system and watch it leak, or not leak, out of the capillaries. I suppose you could put it in the lumen of the G-I or UG tract as well, if that is the tissue you are interested in. I do not know of the specific details of this method, but it has been used since the 1970's, and probably earlier, to show transcapillary transport. Do a google search for "peroxidase tracing" and see what you come up with. Have a look at J. Neurocytology 10:607-643, 1981 for a method using Evans blue as a tracer. It is a small molecule and crosses most endothelia easlily but it might be what you want. I seem to remember that lanthanum has also been used to trace leakage, or lack thereof, across epithelia. Keep in mind that these methods are quite "old" and, while well-documented in the literature, may not show up in an internet search. You may have to go to the library. Geoff Walters, Katherine S wrote: >Hi all, > >I have kind of an interesting problem. We want to look at tight >junctions in cells. We need a small molecule stain that will not be >taken up by the cells. We want the stain to get between the cells and >be stopped by the tight junctions. We do not want to use EM and we do >not want to use fluorescence. Has anybody done this, or have an idea >about how to do this? > >Thank you, >Kathy Walters >Central Microscopy Research Facility >University of Iowa > > >_______________________________________________ >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 glv4708 <@t> bjc.org Thu Jan 13 09:31:31 2005 From: glv4708 <@t> bjc.org (Gretchen Vollmer) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Control Slides for HPV ISH--ThinPrep Message-ID: Good Morning All, I received a nice protocol on making control slides for HPV ISH for SurePath Paps when I posted this question a few weeks ago. Does anyone have a protocol for when ThinPrep (Cytyc) Paps are used? I have the protocol that Ventana sent. I would like to hear from someone who is actually making their own control slides from ThinPrep samples. Thanks in advance, Gretchen Vollmer, CT(ASCP) Cytology Supervisor Christian Hospital NE 11133 Dunn Road St. Louis, MO 63136 314-653-5888, ext. 83044 From pruegg <@t> ihctech.net Thu Jan 13 10:54:51 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] endothelial cell marker other than PECAM In-Reply-To: <41E648E7.28381.233F3E@localhost> Message-ID: <200501131654.j0DGsl2p019039@chip.viawest.net> I use DAKO F8 on rat ffpe tissue using proteinase k digestion for 5 min. with labelled polymer detection. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Margaret Horne Sent: Thursday, January 13, 2005 3:10 AM To: wen eng Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] endothelial cell marker other than PECAM Hi , I haven't started yet , but am about to try von Willebrand's factor ( Factor VIII ). I'll be doing it on rat, first in FFPE to see if it works, and then on osmium fixed Epon embedded. Wish me luck! I chose Factor 8 because of a paper by Ulger et al , Anat Histol Embryol. 2002 Feb:31(1):31-5 where he compares various rat endothelial cell markers : Factor 8 , RECA-1, PECAM-1 , ICAM- 1, OX-43 and ZO-1. I am buying my anti body from Biocare Medical and their tech support suggests trying a pepsin treatment as they use it on the FFPE tissue at 37 C for 5 min. Hope this helps , don't hesitate to email me if you have any more questions, 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfockler <@t> mail1.vcu.edu Thu Jan 13 10:57:24 2005 From: cfockler <@t> mail1.vcu.edu (cfockler@mail1.vcu.edu) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Peloris Tissue Processor Message-ID: <200501131657.j0DGvOf23457@caladan.vcu.edu> Has anyone heard of, or use the Peloris dual retort tissue processor from Vision-Bio? Candyce Fockler Histotechnician Anatomic Pathology From pmarcum <@t> polysciences.com Thu Jan 13 11:04:48 2005 From: pmarcum <@t> polysciences.com (Pamela Marcum) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Peloris Tissue Processor In-Reply-To: <200501131657.j0DGvOf23457@caladan.vcu.edu> Message-ID: <000f01c4f991$fcdc59d0$7f00a8c0@PMARCUM2K> I just went on the internet and typed in Peloris Tissue Processor and got a wealth o information. Granted it is not customer comments however some great pictures and info to ask more questions about. Best Regards, Pamela A Marcum Histology/Microscopy Product Development Manager Polysciences, Inc. 400 Valley Road Warrington, PA 18976 Telephone: 800-523-2575???? Ext. 167 215-343-6484???? Ext. 167 Fax:? 800-343-3291 ???????? 215-343-0214 > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of > cfockler@mail1.vcu.edu > Sent: Thursday, January 13, 2005 11:57 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Peloris Tissue Processor > > > Has anyone heard of, or use the Peloris dual retort tissue processor > from Vision-Bio? > > Candyce Fockler > Histotechnician > Anatomic Pathology > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From vargasb <@t> nova.edu Thu Jan 13 11:16:08 2005 From: vargasb <@t> nova.edu (Bernardo Vargas-Angel) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] decalcification Message-ID: <200501131716.j0DHG7wp025821@pipeline.acast.nova.edu> Could anyone out there suggest a method for rapid acid decalcification that does not interfere with basophilia? Bernardo Vargas-Angel Ph.D. Research Scientist National Coral Reef Institute Nova Southeastern University 8000 N Ocean Drive, Dania Beach, FL, 33004 Voice +(954) 262-3677 Fax +(954) 262-4027 http://www.nova.edu/ocean/vargas-angel/index.html From kbroomal <@t> NEMOURS.ORG Thu Jan 13 11:22:57 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] CD45 Message-ID: <9BCAC308B27CAD4196D8AC9ADF037AAA110A790C@wlmmsx01.nemours.org> Does anyone know of a CD 45 that will work on Rabbit tissue? I think the samples will be coming to us FFPE. Thanks, Kristen Broomall, HT (ASCP) Histotechnology Core Lab Department of Biomedical Research A.I. duPont Hospital for Children NOTICE...This electronic transmission is intended only for the person(s) named. It may contain information that is (i) proprietary to the sender, and/or (ii) privileged, confidential and/or otherwise exempt from disclosure under applicable State and Federal law, including, but not limited to, privacy standards imposed pursuant to the federal Health Insurance Portability and Accountability Act of 1996 (HIPAA). Receipt by anyone other than the named recipient(s) is not a waiver of any applicable privilege. If you received this confidential communication in error, please notify the sender immediately by reply e-mail message and permanently delete the original message from your system. From kbroomal <@t> NEMOURS.ORG Thu Jan 13 11:51:52 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] CD45 Message-ID: <9BCAC308B27CAD4196D8AC9ADF037AAA110A790D@wlmmsx01.nemours.org> I have found 2 mouse anti-rabbit CD45's, one from Research Diagnostics & one from Serotec. Both are indicated for frozen tissue. Does anyone have experience with these? Thanks again! Kristen -----Original Message----- From: Kristen Broomall [mailto:kbroomal@NEMOURS.ORG] Sent: Thursday, January 13, 2005 12:23 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] CD45 Does anyone know of a CD 45 that will work on Rabbit tissue? I think the samples will be coming to us FFPE. Thanks, Kristen Broomall, HT (ASCP) Histotechnology Core Lab Department of Biomedical Research A.I. duPont Hospital for Children NOTICE...This electronic transmission is intended only for the person(s) named. It may contain information that is (i) proprietary to the sender, and/or (ii) privileged, confidential and/or otherwise exempt from disclosure under applicable State and Federal law, including, but not limited to, privacy standards imposed pursuant to the federal Health Insurance Portability and Accountability Act of 1996 (HIPAA). Receipt by anyone other than the named recipient(s) is not a waiver of any applicable privilege. If you received this confidential communication in error, please notify the sender immediately by reply e-mail message and permanently delete the original message from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Jan 13 12:15:58 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] decalcification In-Reply-To: <200501131716.j0DHG7wp025821@pipeline.acast.nova.edu> References: <200501131716.j0DHG7wp025821@pipeline.acast.nova.edu> Message-ID: <6.0.0.22.1.20050113110140.01b66990@gemini.msu.montana.edu> Bernardo, If you control the decalcification using a decalcification endpoint test, you can prevent some loss but strong mineral acids still create some loss of positive hematoxylin staining. If you prevent overexposure aka "overdecalcification" of your fixed calcified tissue to any acid, even formic acid - you can minimize your problem or acid protein hydrolysis will affect nuclear i.e. DNA and RNA staining. I presume, from where you are working, that you are decalcifying coral, sea shells, etc?? Some have used Davidson's fixative which contains acetic acid and actually does some decalcification along with fixation. This is a popular fixative for people in Fish and Wildlife here for their trout, fresh water fish studies. There are ways to speed up decalcification other than just using a different acid decalcifier. Acid, no matter which one, can still damage nuclear staining IF you don't know when your sample is free of calcium. One can use a higher concentration of formic acid, gentler than HCl or nitric, but endpoint determination is still advisable as leaving samples in formic acid will result in the problem you are experiencing. One can use lower concentration of HCl from 10% to 5% or less and it will still be fast. There is no law to say you can't dilute a commercial decalcifier (HCl variety) and usually around 10 - 12% HCl in stock solution. Look at the MSDS and see what HCL concentration is in stock decalcifier and dilute accordingly. Control is still key to avoiding nuclear staining loss. At 10:16 AM 1/13/2005, you wrote: >Could anyone out there suggest a method for rapid acid decalcification that >does not interfere with basophilia? > >Bernardo Vargas-Angel Ph.D. >Research Scientist >National Coral Reef Institute >Nova Southeastern University >8000 N Ocean Drive, Dania Beach, FL, 33004 >Voice +(954) 262-3677 >Fax +(954) 262-4027 >http://www.nova.edu/ocean/vargas-angel/index.html > > >_______________________________________________ >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 dlcowie <@t> prodigy.net Thu Jan 13 12:47:48 2005 From: dlcowie <@t> prodigy.net (Dawn Cowie) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Hirschsprungs Message-ID: <20050113184748.24218.qmail@web81006.mail.yahoo.com> hi netters, Does anyone out there know of a stain that can be performed for Hirschprung's without using acetylcholinesterase? I am trying to eliminate this chemical in our lab. Any response would be greatly appreciated. Thanks, Dawn From Jackie.O'Connor <@t> abbott.com Thu Jan 13 12:54:40 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Hirschsprungs Message-ID: I know a used car parts dealer . . . . . Oh, sorry - I thought you meant Hearse Springs. Sorry. Dawn Cowie Sent by: histonet-bounces@lists.utsouthwestern.edu 01/13/2005 12:47 PM To: histonet cc: Subject: [Histonet] Hirschsprungs hi netters, Does anyone out there know of a stain that can be performed for Hirschprung's without using acetylcholinesterase? I am trying to eliminate this chemical in our lab. Any response would be greatly appreciated. Thanks, Dawn _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Thu Jan 13 13:22:00 2005 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Hirschsprungs Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628001C5C1A7@EXCHANGE1.huntingtonhospital.com> We run an S-100 to stain the ganglion cells. Laurie Colbert -----Original Message----- From: Dawn Cowie [mailto:dlcowie@prodigy.net] Sent: Thursday, January 13, 2005 10:48 AM To: histonet Subject: [Histonet] Hirschsprungs hi netters, Does anyone out there know of a stain that can be performed for Hirschprung's without using acetylcholinesterase? I am trying to eliminate this chemical in our lab. Any response would be greatly appreciated. Thanks, Dawn _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mhorne <@t> upei.ca Thu Jan 13 12:18:28 2005 From: mhorne <@t> upei.ca (Margaret Horne) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] endothelial cell marker other than PECAM In-Reply-To: Message-ID: <41E69143.29808.13DF2C8@localhost> We do an Osmium dissolved in FC-72 ( ie. non aqueous ) fixation and then go straight to 100% EtOH ,then actone as transition to Epon. I am not wedded to Epon but have to stick with the OsO4 + FC- 72 as my fix. Did you try osmium at all? 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 akbitting <@t> geisinger.edu Thu Jan 13 13:47:32 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Antibody to T. pallidum Message-ID: Does anyone do IHC staining for T. pallidum? Where do you buy your antibody? I'm looking for a monoclonal, if possible. Thanks! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From browning <@t> HHSC.CA Thu Jan 13 14:04:59 2005 From: browning <@t> HHSC.CA (Browning Deb) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Hirschsprungs Message-ID: <3AADFB88753AD31189C100902786B91C0E278643@hch_nt_exchange.hhsc.ca> Try calretinin. If you have a ffpe case to start with, do the calretinin stain and then compare it to a frozen section to determine the quantity and position of the positive cells. We tried it with a polymer detection system, the staining was pretty good, but have not pursued this any further. -----Original Message----- From: Dawn Cowie [mailto:dlcowie@prodigy.net] Sent: Thursday, January 13, 2005 1:48 PM To: histonet Subject: [Histonet] Hirschsprungs hi netters, Does anyone out there know of a stain that can be performed for Hirschprung's without using acetylcholinesterase? I am trying to eliminate this chemical in our lab. Any response would be greatly appreciated. Thanks, Dawn _______________________________________________ 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 jhabecke <@t> seattlecca.org Thu Jan 13 15:44:59 2005 From: jhabecke <@t> seattlecca.org (Randolph-Habecker, Julie) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Xylene Substitutes Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAE31@wala01.seattlecca.org> Histonetters, My lab is considering switching to a xylene substitute for tissue processing and other routine use. Someone had mentioned that they tried to switch to a xylene substitute a number of years ago and they had trouble getting a few of their antibodies to work. Has anyone else had a similar problem? If so, what substitute did you try and what antibodies were a problem? 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 bucana <@t> audumla.mdacc.tmc.edu Thu Jan 13 15:45:56 2005 From: bucana <@t> audumla.mdacc.tmc.edu (Corazon D. Bucana) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] cryostat, ergonomics Message-ID: <5.1.1.6.0.20050113152842.00a8b0b8@audumla.mdacc.tmc.edu> We have a 6 year old cryostat that has been used heavily and I am looking for input from experts on a good cryostat that is user friendly and ergonomically designed (?). We had problems with some users having shoulder and back pains after prolonged use of the cryostat. We had our institution's wellness expert come and take a look at how the cryostat is used and essentially he said to get rid of it. So, before we invest money on a new unit we would like to hear from cryostat users in the field. Needless to say we will arrange for demos of different units but any tips or comments beforehand will be most appreciated. Thanks, Cora Bucana From jhabecke <@t> seattlecca.org Thu Jan 13 15:49:11 2005 From: jhabecke <@t> seattlecca.org (Randolph-Habecker, Julie) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Ki-67 MIB-5 on FFPE mouse tissue Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAE32@wala01.seattlecca.org> I have another question. I am trying to get Dako's Ki-67 MIB-5 working on FFPE mouse tissue. I have heard that it works on mouse tissue and it works great on FFPE human tonsil. Does anyone have a great protocol for mouse? 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 John.Garlits <@t> STJUDE.ORG Thu Jan 13 16:09:35 2005 From: John.Garlits <@t> STJUDE.ORG (Garlits, John) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] automated IHC on bone sections? Message-ID: <1E0CC447E59C974CA5C7160D2A2854EC091655E7@SJMEMXMB04.stjude.sjcrh.local> Hello, I am writing to ask if anyone out there has successfully done automated IHC on decalcified, paraffin-embedded sections. I have been staining these for GFP, manually, for some time, but I have found that my IHC results are quite variable. I would like to work out an automated method that might be faster and more consistent. I have submitted sections for automated staining at an in-house facility, but the problem is that the actual bone part of my mouse femur sections always peels off. I use an enzyme-based antigen retrieval method when I do them manually, but I understand that automated stainers typically use a heat-based method. I suspect that the heat-based retrieval is what makes the bone peel off because I have tried a manual version of this and witnessed the same result. Do these automated stainers have the ability to omit the heated retrieval step, or even pause it to introduce an enzyme solution? Thanks, John Garlits St. Jude Children's Research Hospital Memphis, TN From Rcartun <@t> harthosp.org Thu Jan 13 16:20:26 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Antibody to T. pallidum Message-ID: We use BioCare Medical's rabbit polyclonal with excellent results. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Angela Bitting" 01/13/05 02:47PM >>> Does anyone do IHC staining for T. pallidum? Where do you buy your antibody? I'm looking for a monoclonal, if possible. Thanks! Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 23-00 100 N Academy Ave. Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Thu Jan 13 16:22:32 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Ki-67 MIB-5 on FFPE mouse tissue In-Reply-To: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAE32@wala01.seattlecca.org> Message-ID: <000201c4f9be$5ff1a170$76d48a80@AMY> Julie The Ki-67 from Dako that works on mouse tissue is the rat anti-mouse Ki-67 (clone TEC-3), cat # M7249. The MIB-5 works on rat tissue. As far as I am aware these Ki-67 antibodies from dako do not cross react with other species. For Ki-67 I use three different antibodies, one for human, one for mouse and one for rat. I have tried the human on rat and mouse and I know that does not work. I'm assuming the MIB-5 will not work on mouse, plus it's a mouse monoclonal. I normally try to stay away from mouse on mouse if other antibodies are available. I use the rat anti-mouse Ki-67 at 1:50 with steam retrieval. I have images and a more detailed protocol if you would like Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Randolph-Habecker, Julie Sent: Thursday, January 13, 2005 2:49 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Ki-67 MIB-5 on FFPE mouse tissue I have another question. I am trying to get Dako's Ki-67 MIB-5 working on FFPE mouse tissue. I have heard that it works on mouse tissue and it works great on FFPE human tonsil. Does anyone have a great protocol for mouse? 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 From liz <@t> premierlab.com Thu Jan 13 16:29:57 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] automated IHC on bone sections? In-Reply-To: <1E0CC447E59C974CA5C7160D2A2854EC091655E7@SJMEMXMB04.stjude.sjcrh.local> Message-ID: <000c01c4f9bf$697101e0$76d48a80@AMY> John I have run FFPE, formic acid decalcifed joint sections on the Dako autostainer. I try to stick with enzyme digestion for the joint sections. I have ran F4/80, IL-6 and complement C3 with success on the autostainer. The F4/80 and IL-6 require proteinase K digestion and that can be done on the autostainer. I'll e-mail you off the histonet some images. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Garlits, John Sent: Thursday, January 13, 2005 3:10 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] automated IHC on bone sections? Hello, I am writing to ask if anyone out there has successfully done automated IHC on decalcified, paraffin-embedded sections. I have been staining these for GFP, manually, for some time, but I have found that my IHC results are quite variable. I would like to work out an automated method that might be faster and more consistent. I have submitted sections for automated staining at an in-house facility, but the problem is that the actual bone part of my mouse femur sections always peels off. I use an enzyme-based antigen retrieval method when I do them manually, but I understand that automated stainers typically use a heat-based method. I suspect that the heat-based retrieval is what makes the bone peel off because I have tried a manual version of this and witnessed the same result. Do these automated stainers have the ability to omit the heated retrieval step, or even pause it to introduce an enzyme solution? Thanks, John Garlits St. Jude Children's Research Hospital Memphis, TN _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Thu Jan 13 16:51:37 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] SURVEY Message-ID: <83AACDB0810528418AA106F9AE9B7F7E5078C9@sjhaexc02.sjha.org> Two questions for "O ye wise ones" 1. Do you all, especially hospitals, match technical CPT billing codes with your pathologist's professional billing codes? And if so, how do you do this? Who does this? Do your pathologists? Is it done before or after diagnosis? How do you meet the hospital billing window? 2. When your pathologist is paid to perform autopsies by the hospital, does the fee include lab testing or does the hospital pay additional monies to the pathologist to cover cost of immunos, special stains, cultutes, drug screens, etc.? As always, thanks for your help. j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s 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 mucram11 <@t> comcast.net Thu Jan 13 17:33:30 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] mucram11@earthlink.net has a new email address Message-ID: <19003102.1105659210949.JavaMail.vmail@service2.colo.trueswitch.com> mucram11@earthlink.net Mail Account has a new email address Hello, I have just switched my e-mail address from mucram11@earthlink.net to mucram11@comcast.net. Please use this new address for all future e-mails and instant messages. Switching was easy using EasyChange's automatic Internet account switching service. You should check it out! Thanks, mucram11@comcast.net Note: This message was sent at the request of [1]mucram11@comcast.net [2][USEMAP:btm_sign.gif] References 1. mailto:mucram11@comcast.net 2. LYNXIMGMAP:file://localhost/tmp/@29187.1.html#Map From gcallis <@t> montana.edu Thu Jan 13 17:50:48 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Re:cryostat, ergonomics In-Reply-To: <5.1.1.6.0.20050113152842.00a8b0b8@audumla.mdacc.tmc.edu> References: <5.1.1.6.0.20050113152842.00a8b0b8@audumla.mdacc.tmc.edu> Message-ID: <6.0.0.22.1.20050113163814.01b46348@gemini.msu.montana.edu> WE have 3 Leica 1850, but we work at cryostats for hours using a pneumatic chair to adjust height to accomodate for tall or short technicians. If you have to do any weird reaching for controls inside or outside the cryostat, watch out, they should be within easy reach of fingertips. Check for easy reach to flywheel and any strange angled position of fingers, thumb or wrist when working touchpads is uncomfortable. Go for semi automated or automated and reevaluate how you work at a cryostat that adds to back and shoulder problems. Ergonomics is more than just a new cryostat, but how you operate, sit, bend even at the new ones. The variable height chair has proven invaluable, not just some funky cheap stool, or bending over the chamber - seat yourself comfortably and it improves access to all controls. At 02:45 PM 1/13/2005, you wrote: >We have a 6 year old cryostat that has been used heavily and I am looking >for input from experts on a good cryostat that is user friendly and >ergonomically designed (?). We had problems with some users having >shoulder and back pains after prolonged use of the cryostat. We had our >institution's wellness expert come and take a look at how the cryostat is >used and essentially he said to get rid of it. So, before we >invest money on a new unit we would like to hear from cryostat users in >the field. Needless to say we will arrange for demos of different units >but any tips or comments beforehand will be most appreciated. > >Thanks, > >Cora Bucana > > >_______________________________________________ >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 Kemlo.Rogerson <@t> elht.nhs.uk Fri Jan 14 02:26:11 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] decalcification[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EE56@bhrv-nt-11.bhrv.nwest.nhs.uk> EDTA? Not as rapid as you would expect but does not interfere with basophilia, depends which is the main controlling parameter. -----Original Message----- From: Bernardo Vargas-Angel [mailto:vargasb@nova.edu] Sent: 13 January 2005 17:16 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] decalcification[Scanned] Could anyone out there suggest a method for rapid acid decalcification that does not interfere with basophilia? Bernardo Vargas-Angel Ph.D. Research Scientist National Coral Reef Institute Nova Southeastern University 8000 N Ocean Drive, Dania Beach, FL, 33004 Voice +(954) 262-3677 Fax +(954) 262-4027 http://www.nova.edu/ocean/vargas-angel/index.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Fri Jan 14 02:30:29 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] decalcification[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EE57@bhrv-nt-11.bhrv.nwest.nhs.uk> Ergo, EDTA. If the main desire is to retain staining integrity, then anything containing an acid will compromise that. Never tried using EDTA at raised temperatures, that may increase the rate of decalcification; depends on the size of the lump too. -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: 13 January 2005 18:16 To: Bernardo Vargas-Angel; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] decalcification[Scanned] Bernardo, If you control the decalcification using a decalcification endpoint test, you can prevent some loss but strong mineral acids still create some loss of positive hematoxylin staining. If you prevent overexposure aka "overdecalcification" of your fixed calcified tissue to any acid, even formic acid - you can minimize your problem or acid protein hydrolysis will affect nuclear i.e. DNA and RNA staining. I presume, from where you are working, that you are decalcifying coral, sea shells, etc?? Some have used Davidson's fixative which contains acetic acid and actually does some decalcification along with fixation. This is a popular fixative for people in Fish and Wildlife here for their trout, fresh water fish studies. There are ways to speed up decalcification other than just using a different acid decalcifier. Acid, no matter which one, can still damage nuclear staining IF you don't know when your sample is free of calcium. One can use a higher concentration of formic acid, gentler than HCl or nitric, but endpoint determination is still advisable as leaving samples in formic acid will result in the problem you are experiencing. One can use lower concentration of HCl from 10% to 5% or less and it will still be fast. There is no law to say you can't dilute a commercial decalcifier (HCl variety) and usually around 10 - 12% HCl in stock solution. Look at the MSDS and see what HCL concentration is in stock decalcifier and dilute accordingly. Control is still key to avoiding nuclear staining loss. At 10:16 AM 1/13/2005, you wrote: >Could anyone out there suggest a method for rapid acid decalcification that >does not interfere with basophilia? > >Bernardo Vargas-Angel Ph.D. >Research Scientist >National Coral Reef Institute >Nova Southeastern University >8000 N Ocean Drive, Dania Beach, FL, 33004 >Voice +(954) 262-3677 >Fax +(954) 262-4027 >http://www.nova.edu/ocean/vargas-angel/index.html > > >_______________________________________________ >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 immrstambo <@t> hotmail.com Fri Jan 14 06:40:16 2005 From: immrstambo <@t> hotmail.com (Christine Tambasco) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] SURVEY In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E5078C9@sjhaexc02.sjha.org> Message-ID: We have a small hospital that uses Meditech computer software. When we enter the specimen it is automatically connected to the CPT code and we daily print the patient billing list that pulls up that code. We do alter it after reading the final diagnosis if it turns out to be incorrect or to add special stain CPT codes. Our turn around time is typically 1 day and the billing dept automatically gets the codes thru the computer. Our Pathologist is not paid for hospital autopsies as he is paid just to be the laboratory director. The county, however, pays for coroners cases and no extra fees are charged by the hospital. It's just a courtesy thing I quess. Good Luck! Christine Tambasco, HT (ASCP) St. Mary's Hospital 427 Guy Park Ave. Amsterdam, New York 12010 ph-5188417287 From: "Weems, Joyce" >To: "Histonet" >Subject: [Histonet] SURVEY >Date: Thu, 13 Jan 2005 17:51:37 -0500 > >Two questions for "O ye wise ones" > >1. Do you all, especially hospitals, match technical CPT billing codes >with your pathologist's professional billing codes? And > if so, how do you do this? Who does this? Do your pathologists? >Is it done before or after diagnosis? How do you meet the > hospital billing window? > > > > 2. When your pathologist is paid to perform autopsies by the hospital, >does the fee include lab testing or does the hospital > > pay additional monies to the pathologist to cover cost of >immunos, special stains, cultutes, drug screens, etc.? > > > >As always, thanks for your help. j:>) > > > >Joyce Weems > >Pathology Manager >Saint Joseph's Hospital >5665 Peachtree Dunwoody Rd NE >Atlanta, GA 30342 >404-851-7376 - Phone >404-851-7831 - Fax > > > > > >Confidentiality Notice ** The information contained in this message may >be privileged and is confidential information intended for the use of >the addressee listed above. If you are neither the intended recipient >nor the employee or agent responsible for delivering this message to the >intended recipient, you are hereby notified that any disclosure, >copying, distribution or the taking of any action in reliance on the >contents of this information is strictly prohibited. If you have >received this communication in error, please notify us immediately by >replying to the message and deleting it from your computer. >Thank you. Saint Josephâs Health System, Inc. > ><< disclaimer.txt >> >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From immrstambo <@t> hotmail.com Fri Jan 14 06:46:46 2005 From: immrstambo <@t> hotmail.com (Christine Tambasco) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] Xylene Substitutes In-Reply-To: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAE31@wala01.seattlecca.org> Message-ID: Hello histonetters- Once again I will offer some information. I use Clear Rite 3 from Richard Allen and havent had any problems with my immunos. I use the Biogenex kit and antibodies. Maybe its the type of antibodies you are using. It has been my experience that you have to be a detective to be in this line of work! Enjoy troubleshooting! Have a good day everyone. Christine Tambasco, HT (ASCP) St. Mary's Hospital 427 Guy Park Ave Amsterdam, New York 12010 phone 5188417287 >From: "Randolph-Habecker, Julie" >To: "'histonet@lists.utsouthwestern.edu'" >Subject: [Histonet] Xylene Substitutes >Date: Thu, 13 Jan 2005 13:44:59 -0800 > >Histonetters, > >My lab is considering switching to a xylene substitute for tissue processing >and other routine use. Someone had mentioned that they tried to switch to a >xylene substitute a number of years ago and they had trouble getting a few >of their antibodies to work. Has anyone else had a similar problem? If so, >what substitute did you try and what antibodies were a problem? > >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 From RBARNHART <@t> summithealth.org Fri Jan 14 07:39:22 2005 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:29 2005 Subject: [Histonet] SURVEY Message-ID: We have billing explosion codes built into our computer system (Meditech) which contains both the technical and professional changes, the code explodes out into 2 charges once it hits the billing side. As the pathologist signs out the case he enters the correct mnemonic for the explosion code which crosses over to the billing side at midnight. We have like 3 days to post charges before the bill drops and we have an average of a 24 hour turnaround time. We only do a few autopsies and usually the pathologist does not request any additional testing. Usually it is a patient that has died in the hospital and all the lab test are already done and if there are special stains or immuno needed the hospital pays for them. >>> "Weems, Joyce" 1/13/2005 5:51:37 PM >>> Two questions for "O ye wise ones" 1. Do you all, especially hospitals, match technical CPT billing codes with your pathologist's professional billing codes? And if so, how do you do this? Who does this? Do your pathologists? Is it done before or after diagnosis? How do you meet the hospital billing window? 2. When your pathologist is paid to perform autopsies by the hospital, does the fee include lab testing or does the hospital pay additional monies to the pathologist to cover cost of immunos, special stains, cultutes, drug screens, etc.? As always, thanks for your help. j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From dmccaig <@t> ckha.on.ca Fri Jan 14 08:13:59 2005 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Control block for copper Message-ID: <3E5A3F039F0BD8118B4700C00D00202404342F@CKHA9> Does anyone have a positive control block they could spare to demonstrate copper using the rhodanine method. This would be truly appreciated. Diana McCaig, MLT Charge Tech, Histology Laboratory Chatham Kent Health Alliance 519-352-6401 (6604) From hymclab <@t> hyhc.com Fri Jan 14 09:34:11 2005 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Xylene Substitutes Message-ID: We have used Histosolve from Thermo Electron (formerly Thermo Shandon) for over 10 years for all of our processing and staining. We have had no troubles with any applications including our immunos. The only thing we use real xylene for anymore is the purge cycles on our processors. Dawn -----Original Message----- From: Randolph-Habecker, Julie [mailto:jhabecke@seattlecca.org] Sent: Thursday, January 13, 2005 3:45 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Xylene Substitutes Histonetters, My lab is considering switching to a xylene substitute for tissue processing and other routine use. Someone had mentioned that they tried to switch to a xylene substitute a number of years ago and they had trouble getting a few of their antibodies to work. Has anyone else had a similar problem? If so, what substitute did you try and what antibodies were a problem? 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 From Stephen.J.Scholz <@t> osfhealthcare.org Fri Jan 14 09:40:33 2005 From: Stephen.J.Scholz <@t> osfhealthcare.org (Scholz, Stephen J.) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] cross contamination Message-ID: <7F1312711CA7474A89B3DF8BA0BA54D0F5F17C@pmc-rfd-mx01.intranet.osfnet.org> Hello all; I have a question for the masses regarding cross contamination in surgical specimens. I would like know how others are handling situations when a small fragment from one specimen gets embedded with a different case. (probably stuck on forceps) When it is obvious upon reading the slide that the fragment doesn't belong does the Histologist remove it? Does the Pathologist comment in the Path Report and what is the common language used (debris, cross-contaminate, ect)? What is done from the Pathologist perspective when the contaminate tissue is similar but logic dictates that it doesn't belong with that case. Again, is it mentioned in the report and what language is used to state the Pathologist believes there is incorrect tissue fragments with the case? I eagerly await your replies, Stephen J. Scholz HT(ASCP) From meyenhmf <@t> umdnj.edu Fri Jan 14 10:05:49 2005 From: meyenhmf <@t> umdnj.edu (meyenhmf@umdnj.edu) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] cryostat, ergonomics In-Reply-To: <5.1.1.6.0.20050113152842.00a8b0b8@audumla.mdacc.tmc.edu> References: <5.1.1.6.0.20050113152842.00a8b0b8@audumla.mdacc.tmc.edu> Message-ID: <1105718749.41e7eddd36051@webmail.umdnj.edu> The only cryostats ever produced (or designed) for a prolonged use without breaking your back was by Dittes-Duspiva/Lide in Germany in the 60ties (I have used them for years, daily! there is a picture in the older Pearse Histochemistry Book). Maybe Slee or Bright has also some advantages over the "bad designs"> I always wonderd why nobody ever came up with a better design then this "bend over the cryostat, braking your back, into the top" type design! So 30 years ago, I designed an ergonomic cryostat, but space, funds and other interests prevented me from pursuing this venture. (Hello manufacturers, I am available!!!) Regards, Markus F. Meyenhofer Quoting "Corazon D. Bucana" : > We have a 6 year old cryostat that has been used heavily and I am looking > > for input from experts on a good cryostat that is user friendly and > ergonomically designed (?). We had problems with some users having > shoulder and back pains after prolonged use of the cryostat. We had our > > institution's wellness expert come and take a look at how the cryostat is > > used and essentially he said to get rid of it. So, before we > invest money on a new unit we would like to hear from cryostat users in > > the field. Needless to say we will arrange for demos of different units > > but any tips or comments beforehand will be most appreciated. > > Thanks, > > Cora Bucana > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- From jengirl1014 <@t> yahoo.com Fri Jan 14 10:28:47 2005 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] re: xylene substitutes Message-ID: <20050114162847.13429.qmail@web60603.mail.yahoo.com> My lab uses Clear Rite 3 from Richard Allan Scientific. I haven't heard any complaints about IHC done on there slides, just compliments that the slides turned out great. If you call them, they can send you out a free sample to test it to see if you like it. I called and got 4 gallons for free!!! It was worth it too. Now I buy it all the time and even use it for staining! I get really nice H&E and other simple stains. Fell free to e-mail me for more info! 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!? Take Yahoo! Mail with you! Get it on your mobile phone. From Terry.Marshall <@t> rothgen.nhs.uk Fri Jan 14 10:36:08 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] cross contamination Message-ID: This is my practise. It is not held out to be perfect, but is honest. If it's very obvious, as in a case yesterday with a colonic gland sticking onto the surface of a piece of skin, I ignore it in all respects. If less than an expert might misconceive it, I mention it. E.g. "a fragment of endometrium, which is clearly a cross-over from another patient is noted." The crunch comes when you suspect it might be from another patient but you can't know it. There is no easy way out of this one - you have to tell it as it is. Then wait for the "can't you do a test?" - "would immunochemistry help?" - "can you do DNA testing" - "what do I tell the patient" and dozens more possible witless comments or questions. As to the language, cross-over or cross contaminant seems to cover any of cutting board, processing and water bath contamination. If you can see it in the block you can be more specific, but there is little point to being so. Luckily, the bad scenario happens infrequently. The worst scenario, where the cross-over is not recognised or suspected seems even less frequent, and of course, can only be suspected in retrospect. 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: Scholz, Stephen J. [mailto:Stephen.J.Scholz@osfhealthcare.org] Sent: 14 January 2005 15:41 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cross contamination Hello all; I have a question for the masses regarding cross contamination in surgical specimens. I would like know how others are handling situations when a small fragment from one specimen gets embedded with a different case. (probably stuck on forceps) When it is obvious upon reading the slide that the fragment doesn't belong does the Histologist remove it? Does the Pathologist comment in the Path Report and what is the common language used (debris, cross-contaminate, ect)? What is done from the Pathologist perspective when the contaminate tissue is similar but logic dictates that it doesn't belong with that case. Again, is it mentioned in the report and what language is used to state the Pathologist believes there is incorrect tissue fragments with the case? I eagerly await your replies, Stephen J. Scholz HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MAUGER <@t> email.chop.edu Fri Jan 14 10:38:09 2005 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Re: CROYSTAT-ERGONOMICS Message-ID: Dear Gayle, What kind of pneumatic chair do you use? Where can I look for one? Thanks, Jo >>> "Rebecca Barnhart" 01/14/05 8:39 AM >>> We have billing explosion codes built into our computer system (Meditech) which contains both the technical and professional changes, the code explodes out into 2 charges once it hits the billing side. As the pathologist signs out the case he enters the correct mnemonic for the explosion code which crosses over to the billing side at midnight. We have like 3 days to post charges before the bill drops and we have an average of a 24 hour turnaround time. We only do a few autopsies and usually the pathologist does not request any additional testing. Usually it is a patient that has died in the hospital and all the lab test are already done and if there are special stains or immuno needed the hospital pays for them. >>> "Weems, Joyce" 1/13/2005 5:51:37 PM >>> Two questions for "O ye wise ones" 1. Do you all, especially hospitals, match technical CPT billing codes with your pathologist's professional billing codes? And if so, how do you do this? Who does this? Do your pathologists? Is it done before or after diagnosis? How do you meet the hospital billing window? 2. When your pathologist is paid to perform autopsies by the hospital, does the fee include lab testing or does the hospital pay additional monies to the pathologist to cover cost of immunos, special stains, cultutes, drug screens, etc.? As always, thanks for your help. j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From chiggerson <@t> memhosp.com Fri Jan 14 11:31:27 2005 From: chiggerson <@t> memhosp.com (chiggerson@memhosp.com) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Pathology Report Retention in Lab Message-ID: Hi! How long is everyone else keeping the lab copy (hard copy) of the pathology report in the lab files? Original kept in Medical Records indefinitely and lab copy is available on-line but does not include associated items such as drawings, reqs, outside consultation reports, etc.. Thanks, Cindy From RBARNHART <@t> summithealth.org Fri Jan 14 12:09:28 2005 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Pathology Report Retention in Lab Message-ID: We no longer keep a hard copy in the lab. We scan the request, consult report, immuno report, FCM report, cytogenetic report and anything else pertaining to the cases into our computer system. If our computer system goes down (which is a very rare occurrence) we can always go to medical records. Becky >>> 1/14/2005 12:31:27 PM >>> Hi! How long is everyone else keeping the lab copy (hard copy) of the pathology report in the lab files? Original kept in Medical Records indefinitely and lab copy is available on-line but does not include associated items such as drawings, reqs, outside consultation reports, etc.. Thanks, Cindy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Jan 14 12:35:16 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Pathology Report Retention in Lab Message-ID: <83AACDB0810528418AA106F9AE9B7F7E5078DD@sjhaexc02.sjha.org> And how long does your Medical Record Dept keep them? j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rebecca Barnhart Sent: Friday, January 14, 2005 1:09 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Pathology Report Retention in Lab We no longer keep a hard copy in the lab. We scan the request, consult report, immuno report, FCM report, cytogenetic report and anything else pertaining to the cases into our computer system. If our computer system goes down (which is a very rare occurrence) we can always go to medical records. Becky >>> 1/14/2005 12:31:27 PM >>> Hi! How long is everyone else keeping the lab copy (hard copy) of the pathology report in the lab files? Original kept in Medical Records indefinitely and lab copy is available on-line but does not include associated items such as drawings, reqs, outside consultation reports, etc.. Thanks, Cindy _______________________________________________ Histonet mailing list Histonet@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 hymclab <@t> hyhc.com Fri Jan 14 13:17:27 2005 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Pathology Report Retention in Lab Message-ID: We only keep the hard copy in lab if it does have an attachment (consult, FS written diagnosis, stone analysis, etc...). If we don't know that it will have a consult until later we just print another one out to attach the outside report to. Last year was the first year we did that and boy did it save room. Dawn -----Original Message----- From: chiggerson@memhosp.com [mailto:chiggerson@memhosp.com] Sent: Friday, January 14, 2005 11:31 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Pathology Report Retention in Lab Hi! How long is everyone else keeping the lab copy (hard copy) of the pathology report in the lab files? Original kept in Medical Records indefinitely and lab copy is available on-line but does not include associated items such as drawings, reqs, outside consultation reports, etc.. Thanks, Cindy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PatPatterson <@t> mhd.com Fri Jan 14 13:37:50 2005 From: PatPatterson <@t> mhd.com (Patterson, Pat) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] FW: MediTech AP Users Message-ID: <293C7C19EFF7D611AE1A0002A53F81140CB43C34@omega.mhd.com> > I'm looking for some contact names of other MediTech IS users for AP lab. > > We have the Client Server system - currently V 5.3 - soon on 5.4. > > Thanks > Pat Patterson > Manager, Pathology > Methodist Dallas Medical Center > pat.patterson@mhd.com > *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. From GDawson <@t> dynacaremilwaukee.com Fri Jan 14 13:44:17 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] endothelial cell marker other than PECAM Message-ID: I have been messing around with a CD200 stain from BD Biosciences that seems to stain endothelium very well. Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI From RBARNHART <@t> summithealth.org Fri Jan 14 13:53:44 2005 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Pathology Report Retention in Lab Message-ID: They are kept indefinitely in medical records. >>> "Weems, Joyce" 1/14/2005 1:35:16 PM >>> And how long does your Medical Record Dept keep them? j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rebecca Barnhart Sent: Friday, January 14, 2005 1:09 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Pathology Report Retention in Lab We no longer keep a hard copy in the lab. We scan the request, consult report, immuno report, FCM report, cytogenetic report and anything else pertaining to the cases into our computer system. If our computer system goes down (which is a very rare occurrence) we can always go to medical records. Becky >>> 1/14/2005 12:31:27 PM >>> Hi! How long is everyone else keeping the lab copy (hard copy) of the pathology report in the lab files? Original kept in Medical Records indefinitely and lab copy is available on-line but does not include associated items such as drawings, reqs, outside consultation reports, etc.. Thanks, Cindy _______________________________________________ Histonet mailing list Histonet@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 info <@t> instrumedics.com Fri Jan 14 14:16:11 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Cross Contamination Message-ID: <013b01c4fa75$ebb40e20$6401a8c0@INSTRUMEDICS22> One of the values of the Cryo-Vac-Away vacuum system for the cryostat is to prevent debris from one block from contaminating a section from a subsequent block. The nozzle at the blockface suctions away the trimming debris as it is generated leaving the knife clean and the cryostat spotless. Bernice Instrumedics www.instrumedics.com From mward <@t> wfubmc.edu Fri Jan 14 14:51:34 2005 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] pathology orders Message-ID: <61135F0455D33347B5AAE209B903A3040BD0EB8C@EXCHVS2.medctr.ad.wfubmc.edu> Recently our clinical labs have gone to electronic orders and no longer receive written or paper requisitions. The tubes are labeled with barcodes which contain all the necessary information for them to receive and act upon the requests. The clinical labs use one type of computer system and we in anatomic pathology use CoPath. We do some tests within our department (Anatomic Pathology) on blood samples (drawn by the phlebotomists) that is orderable on the floors through the clinical lab computer system but are resulted by our Medical Director/Pathologist in CoPath as a pathology report (given a surgical number). It is the policy of our AP department that all specimens must be accompanied by a written requisition, which I assume is standard practice among other institutions. We have recently started having instances where we receive the blood samples but no requisition. I have been trying to explain to the clinical labs our policy and the reasons for it but they keep asking why, if the order is electronic, we need a piece of paper. Currently we are accepting print outs of the electronic order from the computer as our requisition. So far everyone is trying to remember and doing a pretty good job at it, but sometimes they slip up. Are there any other institutions that have a similar situation? Has anyone in Anatomic Pathology gone to electronic ordering of specimens? I would interested in other scenarios like ours. Thanks, and sorry for the length of this posting. Martha Ward, MT (ASCP) QIHC Molecular Diagnostics Lab Wake Forest University Baptist Medical Center From JWEEMS <@t> sjha.org Fri Jan 14 15:09:19 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] pathology orders Message-ID: <83AACDB0810528418AA106F9AE9B7F7E5078E2@sjhaexc02.sjha.org> We accept our non-gyn cytologies electronically, but they are required to send a copy of the transmittal request. Can you pull out the CAP reg in order to document the requirement? Maybe that would help. Good luck, Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Martha Ward Sent: Friday, January 14, 2005 3:52 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] pathology orders Recently our clinical labs have gone to electronic orders and no longer receive written or paper requisitions. The tubes are labeled with barcodes which contain all the necessary information for them to receive and act upon the requests. The clinical labs use one type of computer system and we in anatomic pathology use CoPath. We do some tests within our department (Anatomic Pathology) on blood samples (drawn by the phlebotomists) that is orderable on the floors through the clinical lab computer system but are resulted by our Medical Director/Pathologist in CoPath as a pathology report (given a surgical number). It is the policy of our AP department that all specimens must be accompanied by a written requisition, which I assume is standard practice among other institutions. We have recently started having instances where we receive the blood samples but no requisition. I have been trying to explain to the clinical labs our policy and the reasons for it but they keep asking why, if the order is electronic, we need a piece of paper. Currently we are accepting print outs of the electronic order from the computer as our requisition. So far everyone is trying to remember and doing a pretty good job at it, but sometimes they slip up. Are there any other institutions that have a similar situation? Has anyone in Anatomic Pathology gone to electronic ordering of specimens? I would interested in other scenarios like ours. Thanks, and sorry for the length of this posting. Martha Ward, MT (ASCP) QIHC Molecular Diagnostics Lab Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 chiggerson <@t> memhosp.com Fri Jan 14 15:14:17 2005 From: chiggerson <@t> memhosp.com (chiggerson@memhosp.com) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Pathology Report Retention in Lab In-Reply-To: Message-ID: How long do you keep the hard copy before you pitch/shred it? Cindy hymclab Sent by: "Schneider, Dawn" 01/14/2005 01:17 PM To "'chiggerson@memhosp.com'" , histonet@lists.utsouthwestern.edu cc Subject RE: [Histonet] Pathology Report Retention in Lab We only keep the hard copy in lab if it does have an attachment (consult, FS written diagnosis, stone analysis, etc...). If we don't know that it will have a consult until later we just print another one out to attach the outside report to. Last year was the first year we did that and boy did it save room. Dawn -----Original Message----- From: chiggerson@memhosp.com [mailto:chiggerson@memhosp.com] Sent: Friday, January 14, 2005 11:31 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Pathology Report Retention in Lab Hi! How long is everyone else keeping the lab copy (hard copy) of the pathology report in the lab files? Original kept in Medical Records indefinitely and lab copy is available on-line but does not include associated items such as drawings, reqs, outside consultation reports, etc.. Thanks, Cindy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mickie2507 <@t> netzero.net Sun Jan 16 12:21:22 2005 From: mickie2507 <@t> netzero.net (mickie2507@netzero.net) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Question about IHC Stains on Mohs frozen sections Message-ID: <20050116.102147.10833.175703@webmail10.lax.untd.com> Hello. I would like to solicit responses from anyone currently performing immunohistochemical(IHC) stains on Mohs frozen sections on a routine or semi-routine basis. I am interested in information about antibodies used for melanoma as well as for basal cell or squamous cell carcinomas. This is a relatively new area of use for IHC stains and there is growing interest in the application of these technics for routine use in the clinical Mohs lab. Any of you who know me know I have many years of histology experience including performing IHC on paraffin sections, both manually and by automated IHC stainers. I am interested in use of specific antibodies, including manufacturers and protocols for manual staining. Thank you in advance for taking the time to help me out. Best Regards, Mickie Johnson Mohs Histology Temporary Services Providing Training, Consultation and Vacation Relief to the Mohs Micrographic Surgery community and vacation relief coverage for routine histology. web-site: www.mohshistotemp.com ______________________________________________________________________ Speed up your surfing with NetZero HiSpeed. Now includes pop-up blocker! Only $14.95/month -visit http://www.netzero.com/surf to sign up today! From KDwyer3322 <@t> aol.com Sun Jan 16 15:49:18 2005 From: KDwyer3322 <@t> aol.com (KDwyer3322@aol.com) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] 2005 Texas Society for Histotechnology State Meeting Message-ID: <1ac.2fbe074d.2f1c3b5e@aol.com> To All: TSH 2005 State Meeting Preview Texas Society for Histotechnology 2005 Symposium/Convention Marriott Dallas/Plano at Legacy Town Center 7120 Dallas Parkway, Plano, Texas Thursday, April 21, 2005 - 11:00 a.m. - Golf Outing Tribute Golf Course Friday, April 22, 2005 2:00-5:30 p.m. Workshop 1) HT (ASCP) Examination Readiness - Glenda Hoye Saturday April 23, 2005 8-11:30 A.M. Morning Workshops 2) Quality Issues in Immunohistochemistry - Bryan Hewlett 3) Microwave: The Whole Enchilada - Donna Willis and Jan Minshew 4) Advanced Excellence with Routine H&E Staining and Immunochemical Staining Of Bone Specimens and Bone Marrow Biopsies Following Decalcifications - James Biesecker, M.D. PhD 5) Do You Know What Your Laboratory Workflow Is? - Ms. Ritu Ward Morning Symposiums 1) Immunohistochemistry in the Analysis of Forensic Evidence from a Double Homicide in New Zealand: The Lundy Case - Rodney Miller, M.D. 2) Antibodies As Diagnostic Tools And Their Utility Beyond Diagnostics: Discussion using GI Stromal tumors/KIT as an example - Saime Aksoy, M.D. 3) Keys to Understanding and Applying Academic Articles in the Histopathology Laboratory - Mark Bailey Saturday April 23, 2005 1:00-4:30p.m. Afternoon Workshops 6) Technical Immunohistochemistry: Achieving Reliability and Reproducibility of Immunostains - Rodney Miller, M.D. 7) Is Your Tissue Processor Fighting You? Fight Back - Pam Marcum 8) Preparing for a CAP Inspection - Hector Hernandez and Joe Nocito 9) Color Your World - an Overview of Special Stains - Kim Rhatigan-Drexler Afternoon Symposiums Student Presentations- Texas Histology Schools 4a) - Cancer: A Collection of Diseases - Olukayode Awotesu, Sujatha Kakuru, and Umadevi Narra - MD Anderson Cancer Center and Mary Anderson - Houston Community College 4b) - Special Histologic Techniques - Michael Scabeto - St. Phillips College and Yvonne Koening University of Texas Health Science Center in San Antonio Saturday, April 23, 2005 - 6:30-7:30 p.m. Guest Speaker - Beck Weathers, M.D. Sunday April 24, 2005 8-11:30 A.M. Workshops 10) IHC Mathematics in the Laboratory - Joel Martinez 11) Shipping Diagnostic Specimens- Linda Durbin 12) Quality Standards for Staining - Bryan Hewlett 13) Quality Assurance in the Histology Laboratory - Kathy Dwyer and Debbie Siena Symposiums 5) Team Building 101 - Evelyn Sandberg 6) Pathologists Looks at the Passion & Death of Jesus Christ- Kevin J. McQuaid, M.D. Program Information - Veronica Davis - 972-579-8291 Vendor Information - Judy Webb - 817-927-1024 Golf Outing Information - Donna Willis -817-878-5644 Programs Will Be Mailed in January From jincan <@t> itsa.ucsf.edu Sun Jan 16 23:38:22 2005 From: jincan <@t> itsa.ucsf.edu (jincan) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Question about IHC Stains on Mohs frozen sections Message-ID: <5.1.1.6.0.20050116213733.00b48d50@itsa.ucsf.edu> Dear Mickie, There is an article by Dr. John Zitelli about IHC on frozen mohs sections in Dermatology Surgery, 2004, 30:403-408. Hope it would be of help. Best wishes! James At 06:21 PM 1/16/2005 +0000, you wrote: >Hello. > >I would like to solicit responses from anyone currently performing >immunohistochemical(IHC) stains on Mohs frozen sections on a routine or >semi-routine basis. I am interested in information about antibodies used >for melanoma as well as for basal cell or squamous cell carcinomas. > >This is a relatively new area of use for IHC stains and there is growing >interest in the application of these technics for routine use in the >clinical Mohs lab. > >Any of you who know me know I have many years of histology experience >including performing IHC on paraffin sections, both manually and by >automated IHC stainers. I am interested in use of specific antibodies, >including manufacturers and protocols for manual staining. > >Thank you in advance for taking the time to help me out. > >Best Regards, > >Mickie Johnson >Mohs Histology Temporary Services >Providing Training, Consultation and Vacation Relief to the Mohs >Micrographic Surgery community and vacation relief coverage for routine >histology. >web-site: www.mohshistotemp.com > > >______________________________________________________________________ >Speed up your surfing with NetZero HiSpeed. >Now includes pop-up blocker! >Only $14.95/month -visit http://www.netzero.com/surf to sign up today! > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jincan <@t> itsa.ucsf.edu Sun Jan 16 23:40:31 2005 From: jincan <@t> itsa.ucsf.edu (jincan) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Question about IHC Stains on Mohs frozen sections Message-ID: <5.1.1.6.0.20050116214015.00b48d50@itsa.ucsf.edu> Dear Mickie, There is an article by Dr. John Zitelli about IHC on frozen mohs sections in Dermatology Surgery, 2004, 30:403-408. Hope it would be of help. Best wishes! James At 06:21 PM 1/16/2005 +0000, you wrote: >Hello. > >I would like to solicit responses from anyone currently performing >immunohistochemical(IHC) stains on Mohs frozen sections on a routine or >semi-routine basis. I am interested in information about antibodies used >for melanoma as well as for basal cell or squamous cell carcinomas. > >This is a relatively new area of use for IHC stains and there is growing >interest in the application of these technics for routine use in the >clinical Mohs lab. > >Any of you who know me know I have many years of histology experience >including performing IHC on paraffin sections, both manually and by >automated IHC stainers. I am interested in use of specific antibodies, >including manufacturers and protocols for manual staining. > >Thank you in advance for taking the time to help me out. > >Best Regards, > >Mickie Johnson >Mohs Histology Temporary Services >Providing Training, Consultation and Vacation Relief to the Mohs >Micrographic Surgery community and vacation relief coverage for routine >histology. >web-site: www.mohshistotemp.com > > >______________________________________________________________________ >Speed up your surfing with NetZero HiSpeed. >Now includes pop-up blocker! >Only $14.95/month -visit http://www.netzero.com/surf to sign up today! > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Mon Jan 17 02:26:42 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] RE: Ki-67 MIB-5 on FFPE mouse tissue Message-ID: <4ccf954cc97f.4cc97f4ccf95@amc.uva.nl> Dear Julie and Liz, How about using the anti-Ki67 rabbit monoclonal (clone SP6, LabVision/Neomarkers)? This antibody worked in my hands on human, mouse and rat tissue very well! And you don't have this mouse-on-mouse problem. Just 15 min HIER in EDTA9.0 and a polymer/HRP detection. Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: c.m.vanderloos@amc.uva.nl ----- Original Message ----- >From Elizabeth Chlipala Date Thu, 13 Jan 2005 15:22:32 -0700 To "'Randolph-Habecker, Julie'" , histonet@lists.utsouthwestern.edu Subject RE: [Histonet] Ki-67 MIB-5 on FFPE mouse tissue Julie The Ki-67 from Dako that works on mouse tissue is the rat anti-mouse Ki-67 (clone TEC-3), cat # M7249. The MIB-5 works on rat tissue. As far as I am aware these Ki-67 antibodies from dako do not cross react with other species. For Ki-67 I use three different antibodies, one for human, one for mouse and one for rat. I have tried the human on rat and mouse and I know that does not work. I'm assuming the MIB-5 will not work on mouse, plus it's a mouse monoclonal. I normally try to stay away from mouse on mouse if other antibodies are available. I use the rat anti-mouse Ki-67 at 1:50 with steam retrieval. I have images and a more detailed protocol if you would like Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Randolph-Habecker, Julie Sent: Thursday, January 13, 2005 2:49 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Ki-67 MIB-5 on FFPE mouse tissue I have another question. I am trying to get Dako's Ki-67 MIB-5 working on FFPE mouse tissue. I have heard that it works on mouse tissue and it works great on FFPE human tonsil. Does anyone have a great protocol for mouse? 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 From Kemlo.Rogerson <@t> elht.nhs.uk Mon Jan 17 03:07:26 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] cross contamination[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EE68@bhrv-nt-11.bhrv.nwest.nhs.uk> Do you log it as an incident using IR1? -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 January 2005 16:36 To: Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] This is my practise. It is not held out to be perfect, but is honest. If it's very obvious, as in a case yesterday with a colonic gland sticking onto the surface of a piece of skin, I ignore it in all respects. If less than an expert might misconceive it, I mention it. E.g. "a fragment of endometrium, which is clearly a cross-over from another patient is noted." The crunch comes when you suspect it might be from another patient but you can't know it. There is no easy way out of this one - you have to tell it as it is. Then wait for the "can't you do a test?" - "would immunochemistry help?" - "can you do DNA testing" - "what do I tell the patient" and dozens more possible witless comments or questions. As to the language, cross-over or cross contaminant seems to cover any of cutting board, processing and water bath contamination. If you can see it in the block you can be more specific, but there is little point to being so. Luckily, the bad scenario happens infrequently. The worst scenario, where the cross-over is not recognised or suspected seems even less frequent, and of course, can only be suspected in retrospect. 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: Scholz, Stephen J. [mailto:Stephen.J.Scholz@osfhealthcare.org] Sent: 14 January 2005 15:41 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cross contamination Hello all; I have a question for the masses regarding cross contamination in surgical specimens. I would like know how others are handling situations when a small fragment from one specimen gets embedded with a different case. (probably stuck on forceps) When it is obvious upon reading the slide that the fragment doesn't belong does the Histologist remove it? Does the Pathologist comment in the Path Report and what is the common language used (debris, cross-contaminate, ect)? What is done from the Pathologist perspective when the contaminate tissue is similar but logic dictates that it doesn't belong with that case. Again, is it mentioned in the report and what language is used to state the Pathologist believes there is incorrect tissue fragments with the case? I eagerly await your replies, Stephen J. Scholz HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Mon Jan 17 05:36:20 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] cross contamination[Scanned] Message-ID: I have only done the few cases I consider "iffy". Logging all seems to me a senseless and futile flogging of techs (irrespective of what I "should" do). Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 09:07 To: Marshall Terry Dr, Consultant Histopathologist; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Do you log it as an incident using IR1? -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 January 2005 16:36 To: Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] This is my practise. It is not held out to be perfect, but is honest. If it's very obvious, as in a case yesterday with a colonic gland sticking onto the surface of a piece of skin, I ignore it in all respects. If less than an expert might misconceive it, I mention it. E.g. "a fragment of endometrium, which is clearly a cross-over from another patient is noted." The crunch comes when you suspect it might be from another patient but you can't know it. There is no easy way out of this one - you have to tell it as it is. Then wait for the "can't you do a test?" - "would immunochemistry help?" - "can you do DNA testing" - "what do I tell the patient" and dozens more possible witless comments or questions. As to the language, cross-over or cross contaminant seems to cover any of cutting board, processing and water bath contamination. If you can see it in the block you can be more specific, but there is little point to being so. Luckily, the bad scenario happens infrequently. The worst scenario, where the cross-over is not recognised or suspected seems even less frequent, and of course, can only be suspected in retrospect. 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: Scholz, Stephen J. [mailto:Stephen.J.Scholz@osfhealthcare.org] Sent: 14 January 2005 15:41 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cross contamination Hello all; I have a question for the masses regarding cross contamination in surgical specimens. I would like know how others are handling situations when a small fragment from one specimen gets embedded with a different case. (probably stuck on forceps) When it is obvious upon reading the slide that the fragment doesn't belong does the Histologist remove it? Does the Pathologist comment in the Path Report and what is the common language used (debris, cross-contaminate, ect)? What is done from the Pathologist perspective when the contaminate tissue is similar but logic dictates that it doesn't belong with that case. Again, is it mentioned in the report and what language is used to state the Pathologist believes there is incorrect tissue fragments with the case? I eagerly await your replies, Stephen J. Scholz HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Jan 17 05:56:34 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] cross contamination[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EE6B@bhrv-nt-11.bhrv.nwest.nhs.uk> Depends how you sell it to Staff. You are too intelligent not to understand the benefit of logging errors so that you can detect trends. My feelings are that mistakes are bad enough but if you can learn from them then they have fulfilled at least a positive function. Very hard concept for Medical Staff and BMS to accept; that we all make mistakes, but systems can be changed to trap these errors. But the errors need to be quantified and recorded; I had problems trying to get Staff to accept that they fallible and change their working practices to account for that. Carry over, logically, only occurs at two or three points in the procedure. Not rocket science to find out which bit of the system is failing, is it? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 11:36 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] I have only done the few cases I consider "iffy". Logging all seems to me a senseless and futile flogging of techs (irrespective of what I "should" do). Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 09:07 To: Marshall Terry Dr, Consultant Histopathologist; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Do you log it as an incident using IR1? -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 January 2005 16:36 To: Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] This is my practise. It is not held out to be perfect, but is honest. If it's very obvious, as in a case yesterday with a colonic gland sticking onto the surface of a piece of skin, I ignore it in all respects. If less than an expert might misconceive it, I mention it. E.g. "a fragment of endometrium, which is clearly a cross-over from another patient is noted." The crunch comes when you suspect it might be from another patient but you can't know it. There is no easy way out of this one - you have to tell it as it is. Then wait for the "can't you do a test?" - "would immunochemistry help?" - "can you do DNA testing" - "what do I tell the patient" and dozens more possible witless comments or questions. As to the language, cross-over or cross contaminant seems to cover any of cutting board, processing and water bath contamination. If you can see it in the block you can be more specific, but there is little point to being so. Luckily, the bad scenario happens infrequently. The worst scenario, where the cross-over is not recognised or suspected seems even less frequent, and of course, can only be suspected in retrospect. 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: Scholz, Stephen J. [mailto:Stephen.J.Scholz@osfhealthcare.org] Sent: 14 January 2005 15:41 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cross contamination Hello all; I have a question for the masses regarding cross contamination in surgical specimens. I would like know how others are handling situations when a small fragment from one specimen gets embedded with a different case. (probably stuck on forceps) When it is obvious upon reading the slide that the fragment doesn't belong does the Histologist remove it? Does the Pathologist comment in the Path Report and what is the common language used (debris, cross-contaminate, ect)? What is done from the Pathologist perspective when the contaminate tissue is similar but logic dictates that it doesn't belong with that case. Again, is it mentioned in the report and what language is used to state the Pathologist believes there is incorrect tissue fragments with the case? I eagerly await your replies, Stephen J. Scholz HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Jan 17 06:05:29 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Telepathology[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EE6C@bhrv-nt-11.bhrv.nwest.nhs.uk> Anyone any experience of Telepathology in Cell Path? Specifically the Nikon one using Coolscopes and Eclipse Net. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 From Terry.Marshall <@t> rothgen.nhs.uk Mon Jan 17 06:30:28 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] cross contamination[Scanned] Message-ID: Well Kemlo, I regard carry-over as inevitable, not necessarily a failing. To prevent it you would need new instruments and cutting board for each specimen. Each specimen would need to be processed individually in new reagents. Each specimen would need to be floated out in a cleaned water bath with fresh water. To be sure to be sure, a new forceps would be used for each specimen embedded. In short, the system of block specimen processing, using processing to cover "the lot", is incapable of producing carry-over free results. Were there to be a perceived increased or unacceptable (yes, I know none is "acceptable") amount of carry-over, that is a different matter. Finding out where the carry-over occurred is nothing to do with logging and IR1s. Indeed, probably the commonest place is the water bath. Do *you* in your lab keep a record of who floated out each and every specimen? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 11:57 To: Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Depends how you sell it to Staff. You are too intelligent not to understand the benefit of logging errors so that you can detect trends. My feelings are that mistakes are bad enough but if you can learn from them then they have fulfilled at least a positive function. Very hard concept for Medical Staff and BMS to accept; that we all make mistakes, but systems can be changed to trap these errors. But the errors need to be quantified and recorded; I had problems trying to get Staff to accept that they fallible and change their working practices to account for that. Carry over, logically, only occurs at two or three points in the procedure. Not rocket science to find out which bit of the system is failing, is it? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 11:36 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] I have only done the few cases I consider "iffy". Logging all seems to me a senseless and futile flogging of techs (irrespective of what I "should" do). Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 09:07 To: Marshall Terry Dr, Consultant Histopathologist; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Do you log it as an incident using IR1? -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 January 2005 16:36 To: Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] This is my practise. It is not held out to be perfect, but is honest. If it's very obvious, as in a case yesterday with a colonic gland sticking onto the surface of a piece of skin, I ignore it in all respects. If less than an expert might misconceive it, I mention it. E.g. "a fragment of endometrium, which is clearly a cross-over from another patient is noted." The crunch comes when you suspect it might be from another patient but you can't know it. There is no easy way out of this one - you have to tell it as it is. Then wait for the "can't you do a test?" - "would immunochemistry help?" - "can you do DNA testing" - "what do I tell the patient" and dozens more possible witless comments or questions. As to the language, cross-over or cross contaminant seems to cover any of cutting board, processing and water bath contamination. If you can see it in the block you can be more specific, but there is little point to being so. Luckily, the bad scenario happens infrequently. The worst scenario, where the cross-over is not recognised or suspected seems even less frequent, and of course, can only be suspected in retrospect. 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: Scholz, Stephen J. [mailto:Stephen.J.Scholz@osfhealthcare.org] Sent: 14 January 2005 15:41 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cross contamination Hello all; I have a question for the masses regarding cross contamination in surgical specimens. I would like know how others are handling situations when a small fragment from one specimen gets embedded with a different case. (probably stuck on forceps) When it is obvious upon reading the slide that the fragment doesn't belong does the Histologist remove it? Does the Pathologist comment in the Path Report and what is the common language used (debris, cross-contaminate, ect)? What is done from the Pathologist perspective when the contaminate tissue is similar but logic dictates that it doesn't belong with that case. Again, is it mentioned in the report and what language is used to state the Pathologist believes there is incorrect tissue fragments with the case? I eagerly await your replies, Stephen J. Scholz HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Jan 17 07:06:34 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] cross contamination[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EE6D@bhrv-nt-11.bhrv.nwest.nhs.uk> Obviously, I am as aware of the dangers inherent in producing H&E's, as only up to recently have I been preparing them since I was a pup. Certain carry over I concede, the most desperate being on the processing machine, cannot be eradicated, but that which can, ought. I think my point is that there may a be certain baseline, below which, you may not ever pass. But if you eradicate those you can control, then you can concentrate on those you can't. Innovative changes, I'm sure, can be introduced by the scientific Staff and Manufacturers for the reduction of carry over on machines. As for forceps? What ever happened to the habit of flaming forceps to incinerate carry over on them? I know alas, health and safety!!! Personally I will be having a zero tolerance on carry over and for those of my Staff reading this Listserv, my intentions are as Terry suggests. Both CPA, GLP and the ISO's concentrate on quality management systems and I see no reason why the 'cutter-up', the 'embedder', the 'sectioner', the 'floater out' and the Q/C person are not identified. That was my intention in my last Lab and that is my intention in this. If you don't know who is doing what and when, then how can you audit the system? It's really not that difficult to do, for a good Manager. In fact I'm an advocate of the team system which allows each team the autonomy of producing a finished product that they 'own' and they can Q/C and order up deeper sections, if appropriate, and some special stains previously agreed with the Pathologist. For example PAS's on 'inflammatory' skins, etc. I remain staggered that some BMS's cannot recognise basic tissue and at least some disease processes; maybe you taught me too well Terry, the traits of the master come out in the student, don't they? I agree the water bath is the most common culprit and it accounted for most of the BCC's I managed to get on your sections of a wart. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 12:30 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Well Kemlo, I regard carry-over as inevitable, not necessarily a failing. To prevent it you would need new instruments and cutting board for each specimen. Each specimen would need to be processed individually in new reagents. Each specimen would need to be floated out in a cleaned water bath with fresh water. To be sure to be sure, a new forceps would be used for each specimen embedded. In short, the system of block specimen processing, using processing to cover "the lot", is incapable of producing carry-over free results. Were there to be a perceived increased or unacceptable (yes, I know none is "acceptable") amount of carry-over, that is a different matter. Finding out where the carry-over occurred is nothing to do with logging and IR1s. Indeed, probably the commonest place is the water bath. Do *you* in your lab keep a record of who floated out each and every specimen? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 11:57 To: Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Depends how you sell it to Staff. You are too intelligent not to understand the benefit of logging errors so that you can detect trends. My feelings are that mistakes are bad enough but if you can learn from them then they have fulfilled at least a positive function. Very hard concept for Medical Staff and BMS to accept; that we all make mistakes, but systems can be changed to trap these errors. But the errors need to be quantified and recorded; I had problems trying to get Staff to accept that they fallible and change their working practices to account for that. Carry over, logically, only occurs at two or three points in the procedure. Not rocket science to find out which bit of the system is failing, is it? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 11:36 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] I have only done the few cases I consider "iffy". Logging all seems to me a senseless and futile flogging of techs (irrespective of what I "should" do). Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 09:07 To: Marshall Terry Dr, Consultant Histopathologist; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Do you log it as an incident using IR1? -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 January 2005 16:36 To: Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] This is my practise. It is not held out to be perfect, but is honest. If it's very obvious, as in a case yesterday with a colonic gland sticking onto the surface of a piece of skin, I ignore it in all respects. If less than an expert might misconceive it, I mention it. E.g. "a fragment of endometrium, which is clearly a cross-over from another patient is noted." The crunch comes when you suspect it might be from another patient but you can't know it. There is no easy way out of this one - you have to tell it as it is. Then wait for the "can't you do a test?" - "would immunochemistry help?" - "can you do DNA testing" - "what do I tell the patient" and dozens more possible witless comments or questions. As to the language, cross-over or cross contaminant seems to cover any of cutting board, processing and water bath contamination. If you can see it in the block you can be more specific, but there is little point to being so. Luckily, the bad scenario happens infrequently. The worst scenario, where the cross-over is not recognised or suspected seems even less frequent, and of course, can only be suspected in retrospect. 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: Scholz, Stephen J. [mailto:Stephen.J.Scholz@osfhealthcare.org] Sent: 14 January 2005 15:41 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cross contamination Hello all; I have a question for the masses regarding cross contamination in surgical specimens. I would like know how others are handling situations when a small fragment from one specimen gets embedded with a different case. (probably stuck on forceps) When it is obvious upon reading the slide that the fragment doesn't belong does the Histologist remove it? Does the Pathologist comment in the Path Report and what is the common language used (debris, cross-contaminate, ect)? What is done from the Pathologist perspective when the contaminate tissue is similar but logic dictates that it doesn't belong with that case. Again, is it mentioned in the report and what language is used to state the Pathologist believes there is incorrect tissue fragments with the case? I eagerly await your replies, Stephen J. Scholz HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Mon Jan 17 07:18:42 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] cross contamination[Scanned] Message-ID: If it is feasible, the concept of "ownership" of a case is excellent, and likely to lead to friendly competition and hopefully, pride in the finished product. When I worked with Kemlo years ago, we sequestered the skin cases and he was solely responsible for processing them, separate from the rubbish (AKA anything not a skin), and quickly going through them with me on a double header. This was of course, an ideal and highly satisfactory situation - but I suspect something of a luxury. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 13:07 To: Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Obviously, I am as aware of the dangers inherent in producing H&E's, as only up to recently have I been preparing them since I was a pup. Certain carry over I concede, the most desperate being on the processing machine, cannot be eradicated, but that which can, ought. I think my point is that there may a be certain baseline, below which, you may not ever pass. But if you eradicate those you can control, then you can concentrate on those you can't. Innovative changes, I'm sure, can be introduced by the scientific Staff and Manufacturers for the reduction of carry over on machines. As for forceps? What ever happened to the habit of flaming forceps to incinerate carry over on them? I know alas, health and safety!!! Personally I will be having a zero tolerance on carry over and for those of my Staff reading this Listserv, my intentions are as Terry suggests. Both CPA, GLP and the ISO's concentrate on quality management systems and I see no reason why the 'cutter-up', the 'embedder', the 'sectioner', the 'floater out' and the Q/C person are not identified. That was my intention in my last Lab and that is my intention in this. If you don't know who is doing what and when, then how can you audit the system? It's really not that difficult to do, for a good Manager. In fact I'm an advocate of the team system which allows each team the autonomy of producing a finished product that they 'own' and they can Q/C and order up deeper sections, if appropriate, and some special stains previously agreed with the Pathologist. For example PAS's on 'inflammatory' skins, etc. I remain staggered that some BMS's cannot recognise basic tissue and at least some disease processes; maybe you taught me too well Terry, the traits of the master come out in the student, don't they? I agree the water bath is the most common culprit and it accounted for most of the BCC's I managed to get on your sections of a wart. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 12:30 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Well Kemlo, I regard carry-over as inevitable, not necessarily a failing. To prevent it you would need new instruments and cutting board for each specimen. Each specimen would need to be processed individually in new reagents. Each specimen would need to be floated out in a cleaned water bath with fresh water. To be sure to be sure, a new forceps would be used for each specimen embedded. In short, the system of block specimen processing, using processing to cover "the lot", is incapable of producing carry-over free results. Were there to be a perceived increased or unacceptable (yes, I know none is "acceptable") amount of carry-over, that is a different matter. Finding out where the carry-over occurred is nothing to do with logging and IR1s. Indeed, probably the commonest place is the water bath. Do *you* in your lab keep a record of who floated out each and every specimen? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 11:57 To: Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Depends how you sell it to Staff. You are too intelligent not to understand the benefit of logging errors so that you can detect trends. My feelings are that mistakes are bad enough but if you can learn from them then they have fulfilled at least a positive function. Very hard concept for Medical Staff and BMS to accept; that we all make mistakes, but systems can be changed to trap these errors. But the errors need to be quantified and recorded; I had problems trying to get Staff to accept that they fallible and change their working practices to account for that. Carry over, logically, only occurs at two or three points in the procedure. Not rocket science to find out which bit of the system is failing, is it? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 11:36 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] I have only done the few cases I consider "iffy". Logging all seems to me a senseless and futile flogging of techs (irrespective of what I "should" do). Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 09:07 To: Marshall Terry Dr, Consultant Histopathologist; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Do you log it as an incident using IR1? -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 January 2005 16:36 To: Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] This is my practise. It is not held out to be perfect, but is honest. If it's very obvious, as in a case yesterday with a colonic gland sticking onto the surface of a piece of skin, I ignore it in all respects. If less than an expert might misconceive it, I mention it. E.g. "a fragment of endometrium, which is clearly a cross-over from another patient is noted." The crunch comes when you suspect it might be from another patient but you can't know it. There is no easy way out of this one - you have to tell it as it is. Then wait for the "can't you do a test?" - "would immunochemistry help?" - "can you do DNA testing" - "what do I tell the patient" and dozens more possible witless comments or questions. As to the language, cross-over or cross contaminant seems to cover any of cutting board, processing and water bath contamination. If you can see it in the block you can be more specific, but there is little point to being so. Luckily, the bad scenario happens infrequently. The worst scenario, where the cross-over is not recognised or suspected seems even less frequent, and of course, can only be suspected in retrospect. 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: Scholz, Stephen J. [mailto:Stephen.J.Scholz@osfhealthcare.org] Sent: 14 January 2005 15:41 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cross contamination Hello all; I have a question for the masses regarding cross contamination in surgical specimens. I would like know how others are handling situations when a small fragment from one specimen gets embedded with a different case. (probably stuck on forceps) When it is obvious upon reading the slide that the fragment doesn't belong does the Histologist remove it? Does the Pathologist comment in the Path Report and what is the common language used (debris, cross-contaminate, ect)? What is done from the Pathologist perspective when the contaminate tissue is similar but logic dictates that it doesn't belong with that case. Again, is it mentioned in the report and what language is used to state the Pathologist believes there is incorrect tissue fragments with the case? I eagerly await your replies, Stephen J. Scholz HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Jan 17 07:32:46 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] cross contamination[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EE6E@bhrv-nt-11.bhrv.nwest.nhs.uk> "This was of course, an ideal and highly satisfactory situation - but I suspect something of a luxury." I'm glad you viewed it as a luxury, I enjoyed it myself too! Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 13:19 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] If it is feasible, the concept of "ownership" of a case is excellent, and likely to lead to friendly competition and hopefully, pride in the finished product. When I worked with Kemlo years ago, we sequestered the skin cases and he was solely responsible for processing them, separate from the rubbish (AKA anything not a skin), and quickly going through them with me on a double header. This was of course, an ideal and highly satisfactory situation - but I suspect something of a luxury. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 13:07 To: Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Obviously, I am as aware of the dangers inherent in producing H&E's, as only up to recently have I been preparing them since I was a pup. Certain carry over I concede, the most desperate being on the processing machine, cannot be eradicated, but that which can, ought. I think my point is that there may a be certain baseline, below which, you may not ever pass. But if you eradicate those you can control, then you can concentrate on those you can't. Innovative changes, I'm sure, can be introduced by the scientific Staff and Manufacturers for the reduction of carry over on machines. As for forceps? What ever happened to the habit of flaming forceps to incinerate carry over on them? I know alas, health and safety!!! Personally I will be having a zero tolerance on carry over and for those of my Staff reading this Listserv, my intentions are as Terry suggests. Both CPA, GLP and the ISO's concentrate on quality management systems and I see no reason why the 'cutter-up', the 'embedder', the 'sectioner', the 'floater out' and the Q/C person are not identified. That was my intention in my last Lab and that is my intention in this. If you don't know who is doing what and when, then how can you audit the system? It's really not that difficult to do, for a good Manager. In fact I'm an advocate of the team system which allows each team the autonomy of producing a finished product that they 'own' and they can Q/C and order up deeper sections, if appropriate, and some special stains previously agreed with the Pathologist. For example PAS's on 'inflammatory' skins, etc. I remain staggered that some BMS's cannot recognise basic tissue and at least some disease processes; maybe you taught me too well Terry, the traits of the master come out in the student, don't they? I agree the water bath is the most common culprit and it accounted for most of the BCC's I managed to get on your sections of a wart. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 12:30 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Well Kemlo, I regard carry-over as inevitable, not necessarily a failing. To prevent it you would need new instruments and cutting board for each specimen. Each specimen would need to be processed individually in new reagents. Each specimen would need to be floated out in a cleaned water bath with fresh water. To be sure to be sure, a new forceps would be used for each specimen embedded. In short, the system of block specimen processing, using processing to cover "the lot", is incapable of producing carry-over free results. Were there to be a perceived increased or unacceptable (yes, I know none is "acceptable") amount of carry-over, that is a different matter. Finding out where the carry-over occurred is nothing to do with logging and IR1s. Indeed, probably the commonest place is the water bath. Do *you* in your lab keep a record of who floated out each and every specimen? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 11:57 To: Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Depends how you sell it to Staff. You are too intelligent not to understand the benefit of logging errors so that you can detect trends. My feelings are that mistakes are bad enough but if you can learn from them then they have fulfilled at least a positive function. Very hard concept for Medical Staff and BMS to accept; that we all make mistakes, but systems can be changed to trap these errors. But the errors need to be quantified and recorded; I had problems trying to get Staff to accept that they fallible and change their working practices to account for that. Carry over, logically, only occurs at two or three points in the procedure. Not rocket science to find out which bit of the system is failing, is it? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 11:36 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] I have only done the few cases I consider "iffy". Logging all seems to me a senseless and futile flogging of techs (irrespective of what I "should" do). Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 09:07 To: Marshall Terry Dr, Consultant Histopathologist; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Do you log it as an incident using IR1? -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 January 2005 16:36 To: Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] This is my practise. It is not held out to be perfect, but is honest. If it's very obvious, as in a case yesterday with a colonic gland sticking onto the surface of a piece of skin, I ignore it in all respects. If less than an expert might misconceive it, I mention it. E.g. "a fragment of endometrium, which is clearly a cross-over from another patient is noted." The crunch comes when you suspect it might be from another patient but you can't know it. There is no easy way out of this one - you have to tell it as it is. Then wait for the "can't you do a test?" - "would immunochemistry help?" - "can you do DNA testing" - "what do I tell the patient" and dozens more possible witless comments or questions. As to the language, cross-over or cross contaminant seems to cover any of cutting board, processing and water bath contamination. If you can see it in the block you can be more specific, but there is little point to being so. Luckily, the bad scenario happens infrequently. The worst scenario, where the cross-over is not recognised or suspected seems even less frequent, and of course, can only be suspected in retrospect. 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: Scholz, Stephen J. [mailto:Stephen.J.Scholz@osfhealthcare.org] Sent: 14 January 2005 15:41 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cross contamination Hello all; I have a question for the masses regarding cross contamination in surgical specimens. I would like know how others are handling situations when a small fragment from one specimen gets embedded with a different case. (probably stuck on forceps) When it is obvious upon reading the slide that the fragment doesn't belong does the Histologist remove it? Does the Pathologist comment in the Path Report and what is the common language used (debris, cross-contaminate, ect)? What is done from the Pathologist perspective when the contaminate tissue is similar but logic dictates that it doesn't belong with that case. Again, is it mentioned in the report and what language is used to state the Pathologist believes there is incorrect tissue fragments with the case? I eagerly await your replies, Stephen J. Scholz HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Jan 17 08:53:04 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] cross contamination Scanned][Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EE6F@bhrv-nt-11.bhrv.nwest.nhs.uk> Given procedural alertness is the answer, and then who teaches whom to be alert over what? 'Quality Circles' were popular in the UK some years ago and I suppose has been replaced by our learning culture of management. Some time ago when a more 'Scientific' style of management was promulgated as the means of running organisations there had to be a means of effective communication outside of the established lines, so that one group could share their experiences with another, and learn; ergo 'Quality Circles' which was plagiarised from the Japanese Kaizen. Kaizen and Kanban were used to re-engineer the Automobile Industry in the UK. I think we are tending to adopt a more matrix style of management in UK Labs which is based on good communication (in fact its gets gridlocked without it, a risk). My point is, I think, the only way to drive down errors in any process is to adopt a quality management system; that system will have personnel with certain abilities that will share their skills across the matrix for the benefit of the organisation. In the Lab processes should be risk assessed and subject to error logging, the Quality Manager needs to interpret these errors with due regard to these assessments; you can then tell whom to look where and at what. Ultimately if these errors are not attended to you will find yourself with a Clinical Governance issue. Once you have identified those 'risk processes' you can then alter working practices to drive down errors. But only after entering another cycle of risk assessment, change, rewrite standard operational procedures then audit. So please fill in those IR1's! A more scientific method of continuous incremental improvement. Well, that's the theory anyway........ Available for weddings, funerals, etc... Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: McCormick, James [mailto:JMcCormick@schosp.org] Sent: 17 January 2005 13:50 To: Marshall Terry Dr,Consultant Histopathologist; Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination Scanned][Scanned] Stephen, Kemlo and Terry, I have noted the brisk and important dialogue going back and forth as it concerns "cross over" contamination from one specimen to another. Indeed, we all recognize the issue of "error" and the need to have a "zero defect" system. Deming, who is famous for his concept and teaching of management systems that promote "quality" production, defined "quality circles" as a method of promoting dialogue among production units to identify and FIX production problems. We (and include myself), are an ad hoc group in pursuit of that excellence. The problem is best dealt with by understanding the "chain of custody" from patient to slide. The responsibility for "getting it right" is distributed over the entire chain of custody...beginning with the patient. The patient is the "owner" of the "chain", all of the transaction "links" add one to the other, and the longer the chain the greater is the opportunity of failure. FIRST, failure opportunity occurs in the collection site selected by the physician. SECOND, in the labeling, THIRD, in the storage and transport, FOURTH, in the aliquot sampling and the instruments of the grossing station, FIFTH, cassette label and security, SIXTH processing fluids and paraffin, SEVENTH, at the embedding station, EIGHTH, the microtome blade and apparatus, NINTH, the water bath and related instruments, TENTH, the cover slip station and media, ELEVENTH, slide labeling, TWELFTH, ( I can't think of it, but it must be there !) If you wish to see something scary....take up the "droppings" from the bottom of any processing fluid or paraffin and examine them under the microscope. "vegetable soup" of specimens. The remedy is procedural alertness to the links in the chain. Detection is an experienced and alert histotechnonogist or pathologist. In reporting, it is a good idea to identify the contamination on the report. Best procedure ,if possible, is to resample the specimen and repeat the examination. A bit awkward but it will stand up best in COURT. I do not often step up to the bat but this is one of my "pet" concerns. At some future NSH meeting I would enjoy meeting each of you to continue the dialogue. It's worth a lecture demonstration and if invited I might take up that cause ! J.B.McCormick,M.D. FCAP,(father of Tissue-Tek ) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Monday, January 17, 2005 6:30 AM To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Well Kemlo, I regard carry-over as inevitable, not necessarily a failing. To prevent it you would need new instruments and cutting board for each specimen. Each specimen would need to be processed individually in new reagents. Each specimen would need to be floated out in a cleaned water bath with fresh water. To be sure to be sure, a new forceps would be used for each specimen embedded. In short, the system of block specimen processing, using processing to cover "the lot", is incapable of producing carry-over free results. Were there to be a perceived increased or unacceptable (yes, I know none is "acceptable") amount of carry-over, that is a different matter. Finding out where the carry-over occurred is nothing to do with logging and IR1s. Indeed, probably the commonest place is the water bath. Do *you* in your lab keep a record of who floated out each and every specimen? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 11:57 To: Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Depends how you sell it to Staff. You are too intelligent not to understand the benefit of logging errors so that you can detect trends. My feelings are that mistakes are bad enough but if you can learn from them then they have fulfilled at least a positive function. Very hard concept for Medical Staff and BMS to accept; that we all make mistakes, but systems can be changed to trap these errors. But the errors need to be quantified and recorded; I had problems trying to get Staff to accept that they fallible and change their working practices to account for that. Carry over, logically, only occurs at two or three points in the procedure. Not rocket science to find out which bit of the system is failing, is it? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 11:36 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] I have only done the few cases I consider "iffy". Logging all seems to me a senseless and futile flogging of techs (irrespective of what I "should" do). Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 09:07 To: Marshall Terry Dr, Consultant Histopathologist; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Do you log it as an incident using IR1? -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 January 2005 16:36 To: Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] This is my practise. It is not held out to be perfect, but is honest. If it's very obvious, as in a case yesterday with a colonic gland sticking onto the surface of a piece of skin, I ignore it in all respects. If less than an expert might misconceive it, I mention it. E.g. "a fragment of endometrium, which is clearly a cross-over from another patient is noted." The crunch comes when you suspect it might be from another patient but you can't know it. There is no easy way out of this one - you have to tell it as it is. Then wait for the "can't you do a test?" - "would immunochemistry help?" - "can you do DNA testing" - "what do I tell the patient" and dozens more possible witless comments or questions. As to the language, cross-over or cross contaminant seems to cover any of cutting board, processing and water bath contamination. If you can see it in the block you can be more specific, but there is little point to being so. Luckily, the bad scenario happens infrequently. The worst scenario, where the cross-over is not recognised or suspected seems even less frequent, and of course, can only be suspected in retrospect. 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: Scholz, Stephen J. [mailto:Stephen.J.Scholz@osfhealthcare.org] Sent: 14 January 2005 15:41 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cross contamination Hello all; I have a question for the masses regarding cross contamination in surgical specimens. I would like know how others are handling situations when a small fragment from one specimen gets embedded with a different case. (probably stuck on forceps) When it is obvious upon reading the slide that the fragment doesn't belong does the Histologist remove it? Does the Pathologist comment in the Path Report and what is the common language used (debris, cross-contaminate, ect)? What is done from the Pathologist perspective when the contaminate tissue is similar but logic dictates that it doesn't belong with that case. Again, is it mentioned in the report and what language is used to state the Pathologist believes there is incorrect tissue fragments with the case? I eagerly await your replies, Stephen J. Scholz HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet *** Confidentiality 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 HornHV <@t> archildrens.org Mon Jan 17 10:37:37 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] SURVEY Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322BA30@EMAIL.archildrens.org> We also have Meditech and it used bomb codes that match cpt codes for the technical and the professional charges. We do all our charging up front. Corrections are made if necessary. Special stains, immunos, etc...are added later when performed. They too bomb out. We do not bill anyone for autopsies. The staff do not get paid extra for these. 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 Weems, Joyce Sent: Thursday, January 13, 2005 4:52 PM To: Histonet Subject: [Histonet] SURVEY Two questions for "O ye wise ones" 1. Do you all, especially hospitals, match technical CPT billing codes with your pathologist's professional billing codes? And if so, how do you do this? Who does this? Do your pathologists? Is it done before or after diagnosis? How do you meet the hospital billing window? 2. When your pathologist is paid to perform autopsies by the hospital, does the fee include lab testing or does the hospital pay additional monies to the pathologist to cover cost of immunos, special stains, cultutes, drug screens, etc.? As always, thanks for your help. j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. ------------------------------------------------------------------------------ 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 mainlinemohs <@t> yahoo.com Mon Jan 17 14:15:10 2005 From: mainlinemohs <@t> yahoo.com (Michael Lehrer) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Mohs tech needed Message-ID: <20050117201510.11050.qmail@web30903.mail.mud.yahoo.com> I apologize if this is an incorrect use of this forum, but I am unsure how else to reach out to the histotech community. I am starting a Mohs practice in the Philadelphia suburbs and will need a tech to work with me. I am open to bringing on a fully trained Mohs tech, or, helping to reorient a histotech with frozen section experience. I can guarrantee a fun, laid back environment with very stable hours. Salary, benefits, profit sharing, partnerships, etc., are negotiable if someone wants to grow with me. Thanks! --------------------------------- Do you Yahoo!? Yahoo! Mail - now with 250MB free storage. Learn more. From funderwood <@t> mcohio.org Mon Jan 17 14:49:20 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:30 2005 Subject: [BULK] - RE: [Histonet] cross contamination[Scanned] Message-ID: Cleaning forceps with a test tube brush between specimens (or atleast after those likely to have carry over) is effective. Remembering to clean the brush daily. Fred >>> Kemlo Rogerson 01/17/05 08:06AM >>> Obviously, I am as aware of the dangers inherent in producing H&E's, as only up to recently have I been preparing them since I was a pup. Certain carry over I concede, the most desperate being on the processing machine, cannot be eradicated, but that which can, ought. I think my point is that there may a be certain baseline, below which, you may not ever pass. But if you eradicate those you can control, then you can concentrate on those you can't. Innovative changes, I'm sure, can be introduced by the scientific Staff and Manufacturers for the reduction of carry over on machines. As for forceps? What ever happened to the habit of flaming forceps to incinerate carry over on them? I know alas, health and safety!!! Personally I will be having a zero tolerance on carry over and for those of my Staff reading this Listserv, my intentions are as Terry suggests. Both CPA, GLP and the ISO's concentrate on quality management systems and I see no reason why the 'cutter-up', the 'embedder', the 'sectioner', the 'floater out' and the Q/C person are not identified. That was my intention in my last Lab and that is my intention in this. If you don't know who is doing what and when, then how can you audit the system? It's really not that difficult to do, for a good Manager. In fact I'm an advocate of the team system which allows each team the autonomy of producing a finished product that they 'own' and they can Q/C and order up deeper sections, if appropriate, and some special stains previously agreed with the Pathologist. For example PAS's on 'inflammatory' skins, etc. I remain staggered that some BMS's cannot recognise basic tissue and at least some disease processes; maybe you taught me too well Terry, the traits of the master come out in the student, don't they? I agree the water bath is the most common culprit and it accounted for most of the BCC's I managed to get on your sections of a wart. Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 12:30 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Well Kemlo, I regard carry-over as inevitable, not necessarily a failing. To prevent it you would need new instruments and cutting board for each specimen. Each specimen would need to be processed individually in new reagents. Each specimen would need to be floated out in a cleaned water bath with fresh water. To be sure to be sure, a new forceps would be used for each specimen embedded. In short, the system of block specimen processing, using processing to cover "the lot", is incapable of producing carry-over free results. Were there to be a perceived increased or unacceptable (yes, I know none is "acceptable") amount of carry-over, that is a different matter. Finding out where the carry-over occurred is nothing to do with logging and IR1s. Indeed, probably the commonest place is the water bath. Do *you* in your lab keep a record of who floated out each and every specimen? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 11:57 To: Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Depends how you sell it to Staff. You are too intelligent not to understand the benefit of logging errors so that you can detect trends. My feelings are that mistakes are bad enough but if you can learn from them then they have fulfilled at least a positive function. Very hard concept for Medical Staff and BMS to accept; that we all make mistakes, but systems can be changed to trap these errors. But the errors need to be quantified and recorded; I had problems trying to get Staff to accept that they fallible and change their working practices to account for that. Carry over, logically, only occurs at two or three points in the procedure. Not rocket science to find out which bit of the system is failing, is it? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 11:36 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] I have only done the few cases I consider "iffy". Logging all seems to me a senseless and futile flogging of techs (irrespective of what I "should" do). Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 09:07 To: Marshall Terry Dr, Consultant Histopathologist; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Do you log it as an incident using IR1? -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 January 2005 16:36 To: Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] This is my practise. It is not held out to be perfect, but is honest. If it's very obvious, as in a case yesterday with a colonic gland sticking onto the surface of a piece of skin, I ignore it in all respects. If less than an expert might misconceive it, I mention it. E.g. "a fragment of endometrium, which is clearly a cross-over from another patient is noted." The crunch comes when you suspect it might be from another patient but you can't know it. There is no easy way out of this one - you have to tell it as it is. Then wait for the "can't you do a test?" - "would immunochemistry help?" - "can you do DNA testing" - "what do I tell the patient" and dozens more possible witless comments or questions. As to the language, cross-over or cross contaminant seems to cover any of cutting board, processing and water bath contamination. If you can see it in the block you can be more specific, but there is little point to being so. Luckily, the bad scenario happens infrequently. The worst scenario, where the cross-over is not recognised or suspected seems even less frequent, and of course, can only be suspected in retrospect. 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: Scholz, Stephen J. [mailto:Stephen.J.Scholz@osfhealthcare.org] Sent: 14 January 2005 15:41 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cross contamination Hello all; I have a question for the masses regarding cross contamination in surgical specimens. I would like know how others are handling situations when a small fragment from one specimen gets embedded with a different case. (probably stuck on forceps) When it is obvious upon reading the slide that the fragment doesn't belong does the Histologist remove it? Does the Pathologist comment in the Path Report and what is the common language used (debris, cross-contaminate, ect)? What is done from the Pathologist perspective when the contaminate tissue is similar but logic dictates that it doesn't belong with that case. Again, is it mentioned in the report and what language is used to state the Pathologist believes there is incorrect tissue fragments with the case? I eagerly await your replies, Stephen J. Scholz HT(ASCP) _______________________________________________ Histonet mailing list Histonet@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 Cathy.Stevens <@t> HealthONEcares.com Mon Jan 17 15:00:20 2005 From: Cathy.Stevens <@t> HealthONEcares.com (Stevens Cathy) Date: Fri Sep 16 15:24:30 2005 Subject: **SPAM** [Histonet] SURVEY Message-ID: We default charges in Meditech using CPT codes. The pathologist' have access to the billing codes and print copies of reports and bill for the professional charges from that. they also review most of the daily reports for accuracy in diagnosis coding matching the CPT code. As others have said, we make corrections within the 3 day window. If possible. Your second question is one I also have. We never bill for autopsies, just pay to have them performed but I am not sure about the other billing. I was going to check with HCA'S billing helpline. Especially when we have fetal demise with autopsy and cytogenetics requested. -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Thursday, January 13, 2005 3:52 PM To: Histonet Subject: **SPAM** [Histonet] SURVEY Two questions for "O ye wise ones" 1. Do you all, especially hospitals, match technical CPT billing codes with your pathologist's professional billing codes? And if so, how do you do this? Who does this? Do your pathologists? Is it done before or after diagnosis? How do you meet the hospital billing window? 2. When your pathologist is paid to perform autopsies by the hospital, does the fee include lab testing or does the hospital pay additional monies to the pathologist to cover cost of immunos, special stains, cultutes, drug screens, etc.? As always, thanks for your help. j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From Rachael_Emerson <@t> URMC.Rochester.edu Tue Jan 18 09:37:51 2005 From: Rachael_Emerson <@t> URMC.Rochester.edu (Emerson, Rachael) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] endogenous alk phos Message-ID: Hello. Is there another method(s) to block endogenous alkaline phosphotase in mice, on frozen sections, besides levamisole? Thanks!!!!! Rachael L. Emerson Center for Human Genetics and Molecular Pediatric Diseases University of Rochester Medical Center 575 Elmwood Avenue MRBX 1.11301 Rochester, NY 14642 Tel (585) 275-5073 Fax (585) 276-0232 From Liam.Brennan <@t> bll.n-i.nhs.uk Tue Jan 18 09:53:26 2005 From: Liam.Brennan <@t> bll.n-i.nhs.uk (Brennan, Liam) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Double Immuno staining on the ventana NexES immunostainer Message-ID: Netters- Anyone doing double label immunostaining on the Ventana NexES and willing to share their experiences and/or protocols? thanks in anticipation of your help. Liam Brennan Histopathology Dept Belfast City Hospital Northern Ireland From Kristopher.Kalleberg <@t> unilever.com Tue Jan 18 09:56:43 2005 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Fontana-Masson Message-ID: Is it wise to filter FM silver stain to determine if it is possibly contaminated? Thanks. From jkiernan <@t> uwo.ca Tue Jan 18 10:34:54 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] endogenous alk phos References: Message-ID: <41ED3AAE.56BE1B94@uwo.ca> An older method of inhibiting endogenous alkaline phosphatase is to treat the section with Lugol's or Gram's iodine solution. Obviously not highly specific, but does that matter for your purposes? According to Humason's Animal Tissue Techniques, alkaline phosphatase can also be inhibited by EDTA, formaldehyde, KMnO4, cysteine or sodium arsenate. -- ------------------------------- 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 _______________________________ "Emerson, Rachael" wrote: > > Hello. Is there another method(s) to block endogenous alkaline phosphotase > in mice, on frozen sections, besides levamisole? > > Thanks!!!!! > > Rachael L. Emerson > Center for Human Genetics and Molecular Pediatric Diseases > University of Rochester Medical Center > 575 Elmwood Avenue MRBX 1.11301 > Rochester, NY 14642 > > Tel (585) 275-5073 > Fax (585) 276-0232 From pruegg <@t> ihctech.net Tue Jan 18 10:54:50 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] knife stage for Leica Cryocut 1800 Message-ID: <200501181654.j0IGsn2p019358@chip.viawest.net> Folks, I asked this before but got no response. I am in need of a knife stage to hold tungsten carbide or steel blades (not disposables) for my Leica cryostat so I can cut bone. I know these are laying around in drawers not being used since most switched them out for the stage holding disposable blades. I would be willing to purchase this. Thank you, Patsy From Lawrence.Brett <@t> luht.scot.nhs.uk Tue Jan 18 11:02:58 2005 From: Lawrence.Brett <@t> luht.scot.nhs.uk (Brett, Lawrence) Date: Fri Sep 16 15:24:30 2005 Subject: FW: [Histonet] endogenous alk phos Message-ID: <857344E7D8F90240935A288998A89D811E775F@wgh-ex1.luht.scot.nhs.uk> 20% Acetic Acid for 10 minutes does a good job on paraffins, but I've not tried it on frozens. Might be too harsh. It also has the advantage that it blocks ALL alk phos' including intestinal. Lawrence Brett Royal Infirmary, Edinburgh UK -----Original Message----- From: Emerson, Rachael [mailto:Rachael_Emerson@URMC.Rochester.edu] Sent: 18 January 2005 15:38 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] endogenous alk phos Hello. Is there another method(s) to block endogenous alkaline phosphotase in mice, on frozen sections, besides levamisole? Thanks!!!!! Rachael L. Emerson Center for Human Genetics and Molecular Pediatric Diseases University of Rochester Medical Center 575 Elmwood Avenue MRBX 1.11301 Rochester, NY 14642 Tel (585) 275-5073 Fax (585) 276-0232 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** 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 gcallis <@t> montana.edu Tue Jan 18 11:15:24 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] endogenous alk phos In-Reply-To: References: Message-ID: <6.0.0.22.1.20050118101152.01af4120@gemini.msu.montana.edu> What tissues are you working with? Gut? There is a blocker from KPL, a universal blocker that will work, but you use it at beginning of staining, as one uses peroxidase blocking. It also blocks endog peroxidase. I am not sure what it will do to frozen sections - a shorter time for application, but they say it can be used with frozen sections. It is wonderful with paraffin sections and if one uses this blocker, you do NOT use levamisole in the chromogen. The reagent is ready to use, we just recycle dropper bottles for application of drops. At 08:37 AM 1/18/2005, you wrote: >Hello. Is there another method(s) to block endogenous alkaline phosphotase >in mice, on frozen sections, besides levamisole? > >Thanks!!!!! > >Rachael L. Emerson >Center for Human Genetics and Molecular Pediatric Diseases >University of Rochester Medical Center >575 Elmwood Avenue MRBX 1.11301 >Rochester, NY 14642 > >Tel (585) 275-5073 >Fax (585) 276-0232 > > >_______________________________________________ >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 hhawkins <@t> UTMB.EDU Tue Jan 18 13:30:28 2005 From: hhawkins <@t> UTMB.EDU (Hawkins, Hal K.) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Monastral blue Message-ID: <8D6F233E2A5D574B929F3944F3316FD01A58EA@EXCH2K3.utmb.edu> We need to study changes in vascular permeability in animal models using Monastral Blue B dye. Apparently Sigma-Aldrich no longer supplies this pigment. Does anyone know of another source, or would anyone be willing to share/sell us some? Alternatively, is there another colloid that is useful in the same way at the level of both light and electron microscopy? Thanks, Hal Hawkins University of Texas Medical Branch Galveston, Texas hhawkins@utmb.edu From renee'matherly <@t> louisville.edu Fri Jan 14 17:31:52 2005 From: renee'matherly <@t> louisville.edu (Renee C Matherly) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Kentucky Histology Symposium Notice Message-ID: Kentucky Society for Histotechnology 28th Annual Symposium March 18 & 19, 2005 Louisville, Ky For information contact: Renee' Matherly rmath0516@aol.com From JMcCormick <@t> schosp.org Mon Jan 17 07:49:51 2005 From: JMcCormick <@t> schosp.org (McCormick, James) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] cross contamination Scanned] Message-ID: <913FAC2B773C19488E26AE6572180FA5D5CB47@exch01.schosp.org> Stephen, Kemlo and Terry, I have noted the brisk and important dialogue going back and forth as it concerns "cross over" contamination from one specimen to another. Indeed, we all recognize the issue of "error" and the need to have a "zero defect" system. Deming, who is famous for his concept and teaching of management systems that promote "quality" production, defined "quality circles" as a method of promoting dialogue among production units to identify and FIX production problems. We (and include myself), are an ad hoc group in pursuit of that excellence. The problem is best dealt with by understanding the "chain of custody" from patient to slide. The responsibility for "getting it right" is distributed over the entire chain of custody...beginning with the patient. The patient is the "owner" of the "chain", all of the transaction "links" add one to the other, and the longer the chain the greater is the opportunity of failure. FIRST, failure opportunity occurs in the collection site selected by the physician. SECOND, in the labeling, THIRD, in the storage and transport, FOURTH, in the aliquot sampling and the instruments of the grossing station, FIFTH, cassette label and security, SIXTH processing fluids and paraffin, SEVENTH, at the embedding station, EIGHTH, the microtome blade and apparatus, NINTH, the water bath and related instruments, TENTH, the cover slip station and media, ELEVENTH, slide labeling, TWELFTH, ( I can't think of it, but it must be there !) If you wish to see something scary....take up the "droppings" from the bottom of any processing fluid or paraffin and examine them under the microscope. "vegetable soup" of specimens. The remedy is procedural alertness to the links in the chain. Detection is an experienced and alert histotechnonogist or pathologist. In reporting, it is a good idea to identify the contamination on the report. Best procedure ,if possible, is to resample the specimen and repeat the examination. A bit awkward but it will stand up best in COURT. I do not often step up to the bat but this is one of my "pet" concerns. At some future NSH meeting I would enjoy meeting each of you to continue the dialogue. It's worth a lecture demonstration and if invited I might take up that cause ! J.B.McCormick,M.D. FCAP,(father of Tissue-Tek ) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Monday, January 17, 2005 6:30 AM To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Well Kemlo, I regard carry-over as inevitable, not necessarily a failing. To prevent it you would need new instruments and cutting board for each specimen. Each specimen would need to be processed individually in new reagents. Each specimen would need to be floated out in a cleaned water bath with fresh water. To be sure to be sure, a new forceps would be used for each specimen embedded. In short, the system of block specimen processing, using processing to cover "the lot", is incapable of producing carry-over free results. Were there to be a perceived increased or unacceptable (yes, I know none is "acceptable") amount of carry-over, that is a different matter. Finding out where the carry-over occurred is nothing to do with logging and IR1s. Indeed, probably the commonest place is the water bath. Do *you* in your lab keep a record of who floated out each and every specimen? Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 11:57 To: Marshall Terry Dr, Consultant Histopathologist; Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Depends how you sell it to Staff. You are too intelligent not to understand the benefit of logging errors so that you can detect trends. My feelings are that mistakes are bad enough but if you can learn from them then they have fulfilled at least a positive function. Very hard concept for Medical Staff and BMS to accept; that we all make mistakes, but systems can be changed to trap these errors. But the errors need to be quantified and recorded; I had problems trying to get Staff to accept that they fallible and change their working practices to account for that. Carry over, logically, only occurs at two or three points in the procedure. Not rocket science to find out which bit of the system is failing, is it? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 17 January 2005 11:36 To: Kemlo Rogerson; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] I have only done the few cases I consider "iffy". Logging all seems to me a senseless and futile flogging of techs (irrespective of what I "should" do). Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 17 January 2005 09:07 To: Marshall Terry Dr, Consultant Histopathologist; Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] Do you log it as an incident using IR1? -----Original Message----- From: Marshall Terry Dr,Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 14 January 2005 16:36 To: Scholz, Stephen J.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cross contamination[Scanned] This is my practise. It is not held out to be perfect, but is honest. If it's very obvious, as in a case yesterday with a colonic gland sticking onto the surface of a piece of skin, I ignore it in all respects. If less than an expert might misconceive it, I mention it. E.g. "a fragment of endometrium, which is clearly a cross-over from another patient is noted." The crunch comes when you suspect it might be from another patient but you can't know it. There is no easy way out of this one - you have to tell it as it is. Then wait for the "can't you do a test?" - "would immunochemistry help?" - "can you do DNA testing" - "what do I tell the patient" and dozens more possible witless comments or questions. As to the language, cross-over or cross contaminant seems to cover any of cutting board, processing and water bath contamination. If you can see it in the block you can be more specific, but there is little point to being so. Luckily, the bad scenario happens infrequently. The worst scenario, where the cross-over is not recognised or suspected seems even less frequent, and of course, can only be suspected in retrospect. 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: Scholz, Stephen J. [mailto:Stephen.J.Scholz@osfhealthcare.org] Sent: 14 January 2005 15:41 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] cross contamination Hello all; I have a question for the masses regarding cross contamination in surgical specimens. I would like know how others are handling situations when a small fragment from one specimen gets embedded with a different case. (probably stuck on forceps) When it is obvious upon reading the slide that the fragment doesn't belong does the Histologist remove it? Does the Pathologist comment in the Path Report and what is the common language used (debris, cross-contaminate, ect)? What is done from the Pathologist perspective when the contaminate tissue is similar but logic dictates that it doesn't belong with that case. Again, is it mentioned in the report and what language is used to state the Pathologist believes there is incorrect tissue fragments with the case? I eagerly await your replies, Stephen J. Scholz HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet *** Confidentiality 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 liz <@t> premierlab.com Tue Jan 18 13:57:00 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Monastral blue In-Reply-To: <8D6F233E2A5D574B929F3944F3316FD01A58EA@EXCH2K3.utmb.edu> Message-ID: <000801c4fd97$df5dec20$76d48a80@AMY> Hal When we have done vascular permeability studies in the past we have used Evans Blue Dye (30 mg/kg in a volume of 4 ml/kg (mice) or 1 ml/kg (rat)), injected IV when we are using capsaicin in a systemic model or Evans blue dye (1 mg/kg in a volume of 1 ml/kg (rat)) is injected IV and then 20 ?l intradermal injections of test articles. I hope this helps. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Hawkins, Hal K. Sent: Tuesday, January 18, 2005 12:30 PM To: Histonet Subject: [Histonet] Monastral blue We need to study changes in vascular permeability in animal models using Monastral Blue B dye. Apparently Sigma-Aldrich no longer supplies this pigment. Does anyone know of another source, or would anyone be willing to share/sell us some? Alternatively, is there another colloid that is useful in the same way at the level of both light and electron microscopy? Thanks, Hal Hawkins University of Texas Medical Branch Galveston, Texas hhawkins@utmb.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cbono <@t> epl-inc.com Tue Jan 18 14:23:15 2005 From: cbono <@t> epl-inc.com (Cyndi Bono) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Phospholipids stain Message-ID: <002901c4fd9b$8aa5b3d0$f201a8c0@cbono1> I'm looking for a stain for phospholipids with similar results to the Baker's Method. Is there a modified Bakers that has shorter timing? Or an alternative stain? Cyndi Bono Client Services Director EPL, Inc. PO Box 474 Herndon, VA 20172-0474 Tel: 703-471-7060 X221 Fax: 703-471-8447 email: cbono@epl-inc.com From Don.Birgerson <@t> leica-microsystems.com Tue Jan 18 15:05:48 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] knife stage for Leica Cryocut 1800 Message-ID: Hi Patsy, I think you scared people with your reference to carbide tungsten blades. You need a standard steel blade holder. Leica calls these "CN" knife holders. Please feel free to phone if you have further questions. Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 CT "Patsy Ruegg" To: Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] knife stage for Leica Cryocut 1800 western.edu 01/18/2005 10:54 AM Folks, I asked this before but got no response. I am in need of a knife stage to hold tungsten carbide or steel blades (not disposables) for my Leica cryostat so I can cut bone. I know these are laying around in drawers not being used since most switched them out for the stage holding disposable blades. I would be willing to purchase this. Thank you, Patsy _______________________________________________ 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 galinadeyneko <@t> yahoo.com Tue Jan 18 16:14:23 2005 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] heart specimens Message-ID: <20050118221423.25869.qmail@web14525.mail.yahoo.com> Galina Deyneko wrote: Dear colleagues please help! I am performing evaluation of the mouse's heart model of cardiac hypertrophy and infarct, but I ran into some problems with microtome sectioning of paraffin blocks of the hearts. It is first time in my practice and I am upset. After harvesting (without perfusion with BNF) hearts are fixed in 10% BNF (I also tried fixation in Bouin's solution and 4% paraphormaldehyde) for 48 hrs and also the time of fixation was extended to 1 week, dehydrated overnight in 70% Ethanol in Cold Room, dissected in short and long axes, and processed in Shandon Processing Center. Ethanol : 60%- 30min Xylene1- 45min 70%-30 min Xylene2- 45 min 95%-40 min Xylene3- 45 min 95%-40 min Wax1- 45 min 100%-40 min Wax2- 1. hrs 100%-45 min Wax3- 1hrs. 100%-45 min I tried to increase and decrease time of processing with and without vacuum and kept in last wax until next morning. Heart tissues appear very dry; they also crumble during sectioning. Before sectioning I kept blocks on ice tray and I also tried to soak them in soap solution. I wetted them with 1% ammonium hydroxide. For waterbath I use distilled water of 43*C. Tissue sections under objective X2 and X4 look unsatisfactory: muscle fibers are torn in many places, crumbled, with pieces torn off, but the remain tissue section mostly without wrinkles. What is significant is that under higher magnification (X100-X400) the cells and nuclei appear fine and clear, but for the photography and measurement my slides are inadequate. Any advice would be highly appreciated. Sincerely, Galina Deyneko. --------------------------------- Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. --------------------------------- Do you Yahoo!? Read only the mail you want - Yahoo! Mail SpamGuard. From bettina.hutz <@t> orionpharma.com Tue Jan 18 23:12:09 2005 From: bettina.hutz <@t> orionpharma.com (bettina.hutz@orionpharma.com) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] heart specimens Message-ID: <243E1A79AF5D89438C70513DBADAA4F0019E40A1@sfies-exchange1.orionnet.org> Hello, I am working on a similar heart model, and first of all: I replaced the Shandon Processing Center by Sakura Tissue Tek VIP5, and suddenly everything works fine:) Nothing against Shandon, but their Histology equipment sucks a big time! Greetings, Tina -----Original Message----- From: Galina Deyneko [mailto:galinadeyneko@yahoo.com] Sent: Wednesday, January 19, 2005 12:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] heart specimens Galina Deyneko wrote: Dear colleagues please help! I am performing evaluation of the mouse's heart model of cardiac hypertrophy and infarct, but I ran into some problems with microtome sectioning of paraffin blocks of the hearts. It is first time in my practice and I am upset. After harvesting (without perfusion with BNF) hearts are fixed in 10% BNF (I also tried fixation in Bouin's solution and 4% paraphormaldehyde) for 48 hrs and also the time of fixation was extended to 1 week, dehydrated overnight in 70% Ethanol in Cold Room, dissected in short and long axes, and processed in Shandon Processing Center. Ethanol : 60%- 30min Xylene1- 45min 70%-30 min Xylene2- 45 min 95%-40 min Xylene3- 45 min 95%-40 min Wax1- 45 min 100%-40 min Wax2- 1. hrs 100%-45 min Wax3- 1hrs. 100%-45 min I tried to increase and decrease time of processing with and without vacuum and kept in last wax until next morning. Heart tissues appear very dry; they also crumble during sectioning. Before sectioning I kept blocks on ice tray and I also tried to soak them in soap solution. I wetted them with 1% ammonium hydroxide. For waterbath I use distilled water of 43*C. Tissue sections under objective X2 and X4 look unsatisfactory: muscle fibers are torn in many places, crumbled, with pieces torn off, but the remain tissue section mostly without wrinkles. What is significant is that under higher magnification (X100-X400) the cells and nuclei appear fine and clear, but for the photography and measurement my slides are inadequate. Any advice would be highly appreciated. Sincerely, Galina Deyneko. --------------------------------- Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. --------------------------------- Do you Yahoo!? Read only the mail you want - Yahoo! Mail SpamGuard. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ewrona <@t> yahoo.com Tue Jan 18 23:13:05 2005 From: ewrona <@t> yahoo.com (Erin Wrona) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Super Tech Message-ID: <20050119051305.44639.qmail@web52501.mail.yahoo.com> Hi Everyone, Does anyone know have a mailing address, website or phone number for Super Tech? We need to reorder TdT and I can't find any information on them. Thanks in advance, Erin From jkiernan <@t> uwo.ca Tue Jan 18 23:52:48 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Re: Phospholipids stain (Long answer) References: <002901c4fd9b$8aa5b3d0$f201a8c0@cbono1> Message-ID: <41EDF5B0.C802F74@uwo.ca> It depends what kind of phospholipids you want to demonstrate, and what else you want to do with the stained slides. Baker's acid haematein method allows you to dehydrate and make a permanent mount because the chromium-haematein end product is not extracted by organic solvents. The method was shown by Adams, in the 1960s, to stain phosphatidylcholines (lecithins) and sphingomyelins but not other types of phospholipid. Bear in mind that cytomembranes contain many kinds of phospholipid and also other hydrophilic lipids. A histochemical method shows mainly sites of high concentration of membranous material such as myelin sheaths, mitochondria and some pathological inclusions. Baker's acid haematein method does not have to be a _very_ long procedure. Baker's original 4-day chromation (of the block, and then of the sections) is nowadays usually replaced by a 4-hour chromation of the frozen sections at 60C, and this is followed by staining in acid haematein and differentiation in borax-ferricyanide, both for 2h at 37C. That's 8-hours for the whole procedure. Alternatively the chromation can take place overnight at 60C so the rest of the method needs only 4 hours (plus a few extra minutes for the rinses, dehydration, clearing and coverslipping). There are plenty of other staining methods for hydrophilic lipids (a group that includes the phospholipids and indeed most lipids other than fats, waxes, cholesterol and cholesterol esters). Techniques are also available for staining hydrophilic and hydrophobic lipids in contrasting colours. There are various control procedures (solvent extractions and chemical blocking reactions) that can support the contention that a particular method has really stained a certain type of lipid. There is also an excellent, small, cheap paperback called Lipid Histochemistry by Olga Bayliss High, who was Adams's principal collaborator. This book gives all the instructions, most of the explanations, and lots of references. Do not even think of doing lipid staining in a research context before reading most of this book. I think it's been out of print for a few years. If it's not in your library, get "Neurohistochemistry" by CWM Adams (1965). Every library has that one; it came out in the years when universities were spending on books, journals, librarians, professors and scholarships for clever students. Those were the days! Adams (1965) is a big book, but it contains detailed explanations that are still valid. There have been several lipid histochemistry innovations since the 1960s, but this is not a rapidly changing field. It is a field in which you need to understand the mechanisms and limitations of the techniques. Several controls are needed to identify a granule, droplet or smudge with even a broad category of lipids. The information is in the literature but anything on the internet (including this message from me) should be carefully checked against trustworthy sources. Anyone can send a reply to a question. That's why you should never believe any internet answer until you have checked out references to substantial sources such as articles in peer-reviewed scientific journals or printed books. Journals generally have the highest standards. Book publishers (especially for textbooks) now have really stringent reviewing. If you read something in a textbook published since 2002 it was probably the "truth" the time of printing. Book standard were high even in the 1960s when Adams's big fat Neurohistochemistry book hit the market. If you can use thr internet to find Lipid Histochemistry by Olga Bayliss High, buy it. It's a classic. John Kiernan London, Canada ___________________________________________ Cyndi Bono wrote: > > I'm looking for a stain for phospholipids with similar results to the Baker's Method. Is there a modified Bakers that has shorter timing? Or an alternative stain? > > Cyndi Bono > Client Services Director > EPL, Inc. > PO Box 474 > Herndon, VA 20172-0474 > Tel: 703-471-7060 X221 > Fax: 703-471-8447 > email: cbono@epl-inc.com > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ajennings <@t> unmc.edu Wed Jan 19 05:17:54 2005 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] heart specimens Message-ID: Have you Checked the temperatures in your paraffin? From BMolinari <@t> heart.thi.tmc.edu Wed Jan 19 05:42:24 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] contamination/molds Message-ID: While on this subject I was just curious..how often do you clean your molds and how do you clean them? Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 From gu.lang <@t> gmx.at Wed Jan 19 05:52:28 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] contamination/molds Message-ID: -----Urspr?ngliche Nachricht----- Von: Gudrun Lang [mailto:gu.lang@gmx.at] Gesendet: Mittwoch, 19. J?nner 2005 12:50 An: 'Molinari, Betsy' Betreff: AW: [Histonet] contamination/molds We clean our molds daily in a ultrasonic bath. Water with tensid, 35 degree; Paraffin swims on the surface, molds become clean. Gudrun Lang Austria -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Molinari, Betsy Gesendet: Mittwoch, 19. J?nner 2005 12:42 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] contamination/molds While on this subject I was just curious..how often do you clean your molds and how do you clean them? Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Wed Jan 19 06:23:37 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] formic acid / Nasd on bone marrow Message-ID: To those, who use formic acid to decal bone marrow trephines. Have you experience with performing Naphtol-AS-D-chloroacetat-esterase on your slides? Until now we decal with buffered EDTA an our NASD works well. Immunos show some difficulties. So we start to test formic acid. I would like to know, if the esterase is influenced by the acid. Thank you Gudrun Lang Akh Linz, Austria From hodges420 <@t> msn.com Wed Jan 19 09:09:16 2005 From: hodges420 <@t> msn.com (MARY T HODGES) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] lecture to students Message-ID: Good morning all, I need your input again. My doctor is going to do a lecture to the histo students about GI biopsys enbedding ,cutting and staining. If anyone has any articles ect. please let me know Thanks Tere Hodges From Gabriel.Marceau <@t> inrs-iaf.uquebec.ca Wed Jan 19 08:31:52 2005 From: Gabriel.Marceau <@t> inrs-iaf.uquebec.ca (Marceau, Gabriel (IAF)(LAVAL)) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Myelin basic Protein in mouse Message-ID: Ray, Serotec has a Rat anti-MBP under the catalogue number MCA408S but it is a culture supernatant, is this what you are using at the dilution of 1:2000? Usually supernatant are used pure or nearly-pure this is why I ask. If not can you still order number MCA408 even if no longuer in the catalogue? Thanks, Gabriel ------------------------------------------------------------ Gabriel Marceau INRS-Institut Armand-Frappier 531, boul. des Prairies Laval (Qu?bec) H7V 1B7 T?l.: (450) 687-5010 poste 4282 From: "Koelling, Ray" Cynthia, for a couple of years we've run rat-antiMBP from Serotec (MCA408) on mouse brain fixed 24 hours in NBF. At 1:1000-1:2000 overnight at 4 degrees with no retrieval what-so-ever. Then biotinylated rabbit anti rat and so forth. Ray Raymond Koelling Research Scientist, Pathology Amgen Corp. Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Favara, Cynthia (NIH/NIAID) Sent: Friday, January 07, 2005 11:21 AM To: Histonet Subject: [Histonet] Myelin basic Protein in mouse I have run a quick search in the archives and found nothing. Anyone with experience with Myelin Basic Protein immunohistochemistry in FFPE murine tissue? Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From HornHV <@t> archildrens.org Wed Jan 19 08:47:19 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] disposable lab coats Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322BA34@EMAIL.archildrens.org> I am wanting to know how many of you are using disposable lab coats. If you use them, can you estimate how long your techs wear one coat? Do you have a cheap supply source? Thanks! Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 ------------------------------------------------------------------------------ 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 juan.gutierrez <@t> christushealth.org Wed Jan 19 08:50:56 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Super Tech Message-ID: Supertechs, Inc. 9610 Medical Center Drive, Suite 101 Rockville, MD 20850 Ph. (301)309-6695 Fx. (301)294-2451 I could not find a website, but I hope this helps. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Erin Wrona Sent: Tuesday, January 18, 2005 11:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Super Tech Hi Everyone, Does anyone know have a mailing address, website or phone number for Super Tech? We need to reorder TdT and I can't find any information on them. Thanks in advance, Erin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Wed Jan 19 08:58:01 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] disposable lab coats Message-ID: My techs go thru one a week on average. I believe we get them from Fischer, the hospital orders them for us, but I rather have the real thing. They are hot and they rip real easy, plus fluids go through them as if they weren't even there. Just my two cents. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Wednesday, January 19, 2005 8:47 AM To: Histonet Subject: [Histonet] disposable lab coats I am wanting to know how many of you are using disposable lab coats. If you use them, can you estimate how long your techs wear one coat? Do you have a cheap supply source? Thanks! Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From koellinr <@t> amgen.com Wed Jan 19 09:08:12 2005 From: koellinr <@t> amgen.com (Koelling, Ray) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Myelin basic Protein in mouse Message-ID: <16834C6DFFA6004C88DE4507FB8AE544018CC3FC@wa-mb4-sea.amgen.com> Hi Gabriel, I see your dilema. The 2004 Serotec catalog lists MCA408S as a supernatant. Can't find my old catalog and I haven't ordered for a while but my vial is 250 ul of MCA408 (no S) and is batch 050599. It is from Aug 2001 and still working nicely at 1:2000 or so. Maybe a call to company can trace why the #'s are different. Possibly clone died off or drifted or they are trying to re-establish it in vivo??? Ray Raymond Koelling Research Scientist-Pathology Amgen Corp. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marceau, Gabriel (IAF)(LAVAL) Sent: Wednesday, January 19, 2005 6:32 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Myelin basic Protein in mouse Ray, Serotec has a Rat anti-MBP under the catalogue number MCA408S but it is a culture supernatant, is this what you are using at the dilution of 1:2000? Usually supernatant are used pure or nearly-pure this is why I ask. If not can you still order number MCA408 even if no longuer in the catalogue? Thanks, Gabriel ------------------------------------------------------------ Gabriel Marceau INRS-Institut Armand-Frappier 531, boul. des Prairies Laval (Qu?bec) H7V 1B7 T?l.: (450) 687-5010 poste 4282 From: "Koelling, Ray" Cynthia, for a couple of years we've run rat-antiMBP from Serotec (MCA408) on mouse brain fixed 24 hours in NBF. At 1:1000-1:2000 overnight at 4 degrees with no retrieval what-so-ever. Then biotinylated rabbit anti rat and so forth. Ray Raymond Koelling Research Scientist, Pathology Amgen Corp. Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Favara, Cynthia (NIH/NIAID) Sent: Friday, January 07, 2005 11:21 AM To: Histonet Subject: [Histonet] Myelin basic Protein in mouse I have run a quick search in the archives and found nothing. Anyone with experience with Myelin Basic Protein immunohistochemistry in FFPE murine tissue? Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sa.drew <@t> hosp.wisc.edu Wed Jan 19 09:11:40 2005 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Is anyone familiar w/BNP? Message-ID: We have a resident who wanted to know where we could send autopsy slides for brain type naterietic(sp?) peptide...Help please! Thank you! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 From Dndsomi <@t> aol.com Wed Jan 19 09:15:08 2005 From: Dndsomi <@t> aol.com (Dndsomi@aol.com) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Fatty Specimen Message-ID: <02DD4254.6D041FAE.0016AED6@aol.com> Having a problem with breast and colon blocks not being fixed well. We are using Rapid Fix and this has worked well in the past - any suggestions? Thanks, DDietz From Patty.Lott <@t> ORTHO.UAB.EDU Wed Jan 19 09:31:33 2005 From: Patty.Lott <@t> ORTHO.UAB.EDU (Patty Lott) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] H&E on plastic sections Message-ID: <85F6C7A1330E794DB8540AFD001CC77E05328D69@rosco.ortho.uab.edu> Does anyone have a good H&E procedure for thin plastic sections (MMA)? (I think we've been thru this before on histonet, but wanted to check again!) Thanks, Patty Lott, Laboratory Supervisor Orthopaedic Research Laboratory Center for Metabolic Bone Disease Laboratory University of Alabama at Birmingham LHRB B37 0007 1919 7th Ave. South Birmingham, AL 35294 (205) 934-2007 From gcallis <@t> montana.edu Wed Jan 19 09:35:04 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Re: H&E on plastic sections In-Reply-To: <85F6C7A1330E794DB8540AFD001CC77E05328D69@rosco.ortho.uab.e du> References: <85F6C7A1330E794DB8540AFD001CC77E05328D69@rosco.ortho.uab.edu> Message-ID: <6.0.0.22.1.20050119083125.01b3da30@gemini.msu.montana.edu> Hi Patty, This was discussed on Histonet just a couple of months ago, in detail. You have to remove the plastic with 60C xylene so there is NO plastic left in the tissue. Neil Hand did this nicely, and did at least 3 changes to get the MMA out, then rehydrate section and stain - you may have to adjust for your H&E protocol, whatever tweaks are necessary. One nice lady put her staining protocol on 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 Myri37 <@t> aol.com Wed Jan 19 10:00:32 2005 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] staining MMA Message-ID: <0B3F323E.13FB8250.0005167B@aol.com> Hello everyone this is my question about a titane implant embeded with MMA (like always !!!) : I done a thik section with titane and polished to 50um stained with richardson's stain and i see a tissue with no fibers of collagen directed towards titane. and when i remove the titane, and then i can do sections of 5um thikness, staining with goldner trichrome, i see collagen fibers directed towards titane. do you think if it's because of the stain not appropriated with fiber collagen with thik section about 50 um ? Or something else ? is it possible to stain very thin collagen fibers in sections about 50 um thinkness. Thank you very much for any helm and advice. Myriam natural implant France From lyonm <@t> upstate.edu Wed Jan 19 10:16:26 2005 From: lyonm <@t> upstate.edu (Michael Lyon) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] myosin Message-ID: Hi: I am looking for a good marker of developmental myosin that will work in formalin fixed tissues. Most of the antibodies that I have looked at do not work in fixed tissue. This is a problem since it has become very difficult for us to get anything that is unfixed. Any suggestions are welcome. Thanks Michael J. Lyon, Ph.D. Associate Professor of Otolaryngology SUNY Upstate Medical University 156 Weiskotten Hall 750 East Adams Street Syracuse, New York 13210 Voice 315-464-7253 FAX 315-464-5572 From Patty.Lott <@t> ORTHO.UAB.EDU Wed Jan 19 10:24:25 2005 From: Patty.Lott <@t> ORTHO.UAB.EDU (Patty Lott) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] etching? Message-ID: <85F6C7A1330E794DB8540AFD001CC77E05328D6D@rosco.ortho.uab.edu> For you boneheads, what are your favorite methods of etching? Patty Lott, Laboratory Supervisor Orthopaedic Research Laboratory Center for Metabolic Bone Disease Laboratory University of Alabama at Birmingham LHRB B37 0007 1919 7th Ave. South Birmingham, AL 35294 (205) 934-2007 From gcallis <@t> montana.edu Wed Jan 19 10:30:03 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] staining MMA In-Reply-To: <0B3F323E.13FB8250.0005167B@aol.com> References: <0B3F323E.13FB8250.0005167B@aol.com> Message-ID: <6.0.0.22.1.20050119090917.01af5450@gemini.msu.montana.edu> Myriam, What is Richardson's stain? What dyes does it contain? It more than likely is the failure of a dye's (probably of high molecular weight) being able to penetrate MMA plastic in order to stain collagen. There are some ways to help this along, etching with solvents to soften plastic is one way. If it is bone in PMMA, one can 1% formic acid etch for 1 min, rinse well and stain the surface. With thin sections, total removal of plastic probably solves the problem. You should read this publication - Staining of plastic sections: a review of problems, explanations and possible solutions. RW Horobin, J Microscopy 131(2):173-186, 1983. This article will help you understand the problems you encounter and how to try and solve them. PMMA is very hydrophobic, GMA is also hydrophobic but less so than MMA. Dyes with low molecular weight penetrate MMA better than others, these are basic fuchsin, light green, toluidine blue, Stevenels blue or Sandersons rapid bone stain (potassium permangante oxidized methylene blue produces T blue, thionin, azures, etc), the dyes in McNeals tetrachrome, and methylene blue/basic fuchsin comination. High molecular weight dyes probably will require MMA removal, really only possible in a thin section mounted on a slide unless you try solvent etching. Did you remove the MMA from your 5 um thick sections? If so, then dyes can get to the fibers since the MMA is gone. Horobin discusses collagen fibers and how one can reverse colors in trichromes when staining MMA embedded tissues - very interesting chemistry. There are other factors involved, heat, pH, thickness of section. Have you ever tried doing a surface stain of your 50 um sections using McNeals Tetrachrome (do NOT buy this as a commercial solution, make it in house) - use it alone or combine it with toluidine blue as a double stain. Brilliant results. () At 09:00 AM 1/19/2005, you wrote: >Hello everyone >this is my question about a titane implant embeded with MMA (like always >!!!) : >I done a thik section with titane and polished to 50um stained with >richardson's stain and i see a tissue with no fibers of collagen directed >towards titane. >and when i remove the titane, and then i can do sections of 5um thikness, >staining with goldner trichrome, i see collagen fibers directed towards titane. >do you think if it's because of the stain not appropriated with fiber >collagen with thik section about 50 um ? >Or something else ? is it possible to stain very thin collagen fibers in >sections about 50 um thinkness. >Thank you very much for any helm and advice. 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 themagoos <@t> rushmore.com Wed Jan 19 10:33:47 2005 From: themagoos <@t> rushmore.com (Jason and Heather) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] disposable lab coats References: <9AE8AA9E1F644B4AA6C155FB6FD51C6322BA34@EMAIL.archildrens.org> Message-ID: <000f01c4fe44$a8122850$b3fc9fd1@magoo> We use them for about 1-2 weeks at a time. ----- Original Message ----- From: "Horn, Hazel V" To: "Histonet" Sent: Wednesday, January 19, 2005 7:47 AM Subject: [Histonet] disposable lab coats I am wanting to know how many of you are using disposable lab coats. If you use them, can you estimate how long your techs wear one coat? Do you have a cheap supply source? Thanks! Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital Phone - 501.364.4240 Fax - 501.364.3912 ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Gabriel.Marceau <@t> inrs-iaf.uquebec.ca Wed Jan 19 10:49:48 2005 From: Gabriel.Marceau <@t> inrs-iaf.uquebec.ca (Marceau, Gabriel (IAF)(LAVAL)) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Myelin basic Protein in mouse Message-ID: For those it may concern the product in question was sold as an ascite until 2003 (found an old catalogue) and now it is sold only as a culture supernatant. It is the same epitope, same clone, etc. They probably cannot make ascite anymore for the same reasons we can no longuer do that here at the research center. The animal care committee is restricting ascite production. Sadly for the use I need the antibody for it will cost too much to use that product if I cannot dilute it. I want to label myelin on spinal cord sections (cryostat sections of 4%para fixed tissu) but I have "alot" of slides to label... Gabriel ------------------------------------------------------------ Gabriel Marceau INRS-Institut Armand-Frappier 531, boul. des Prairies Laval (Qu?bec) H7V 1B7 -----Original Message----- Sent: Wed 19/01/05 10:08 To: Marceau, Gabriel (IAF)(LAVAL); histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Myelin basic Protein in mouse Hi Gabriel, I see your dilema. The 2004 Serotec catalog lists MCA408S as a supernatant. Can't find my old catalog and I haven't ordered for a while but my vial is 250 ul of MCA408 (no S) and is batch 050599. It is from Aug 2001 and still working nicely at 1:2000 or so. Maybe a call to company can trace why the #'s are different. Possibly clone died off or drifted or they are trying to re-establish it in vivo??? Ray Raymond Koelling Research Scientist-Pathology Amgen Corp. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Marceau, Gabriel (IAF)(LAVAL) Sent: Wednesday, January 19, 2005 6:32 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Myelin basic Protein in mouse Ray, Serotec has a Rat anti-MBP under the catalogue number MCA408S but it is a culture supernatant, is this what you are using at the dilution of 1:2000? Usually supernatant are used pure or nearly-pure this is why I ask. If not can you still order number MCA408 even if no longuer in the catalogue? Thanks, Gabriel ------------------------------------------------------------ Gabriel Marceau INRS-Institut Armand-Frappier 531, boul. des Prairies Laval (Qu?bec) H7V 1B7 T?l.: (450) 687-5010 poste 4282 From: "Koelling, Ray" Cynthia, for a couple of years we've run rat-antiMBP from Serotec (MCA408) on mouse brain fixed 24 hours in NBF. At 1:1000-1:2000 overnight at 4 degrees with no retrieval what-so-ever. Then biotinylated rabbit anti rat and so forth. Ray Raymond Koelling Research Scientist, Pathology Amgen Corp. Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Favara, Cynthia (NIH/NIAID) Sent: Friday, January 07, 2005 11:21 AM To: Histonet Subject: [Histonet] Myelin basic Protein in mouse I have run a quick search in the archives and found nothing. Anyone with experience with Myelin Basic Protein immunohistochemistry in FFPE murine tissue? Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk Wed Jan 19 11:06:07 2005 From: JOHN.PHILLIPS <@t> new-tr.wales.nhs.uk (JOHN PHILLIPS) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] formic acid / Nasd on bone marrow Message-ID: <166A1E642B5B644DA694C08FD29D0ADC603465@ztroy.new-tr.wales.nhs.uk> Esterase activity is lost after acid decalcification. Investigations for naphthol as-d chloroacetate esterase for granulocytes and alpha-naphthyl acetate esterase for monocytes will not work. Both investigations work fine after ADTA decal. John, Wrexham, UK. -----Original Message----- From: Gudrun Lang [mailto:gu.lang@gmx.at] Sent: 19 January 2005 12:24 To: Histonetliste (Histonetliste) Subject: [Histonet] formic acid / Nasd on bone marrow To those, who use formic acid to decal bone marrow trephines. Have you experience with performing Naphtol-AS-D-chloroacetat-esterase on your slides? Until now we decal with buffered EDTA an our NASD works well. Immunos show some difficulties. So we start to test formic acid. I would like to know, if the esterase is influenced by the acid. Thank you Gudrun Lang Akh Linz, Austria _______________________________________________ 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 histo <@t> bthosp.com Wed Jan 19 11:10:08 2005 From: histo <@t> bthosp.com (O'Brien, Sue) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Faxing of Pathology Reports Message-ID: <4D95C24EC0E4C84787A6919F1165B25E24B0A6@btmhems01.BTHOSP.INT> I am looking into how many institutions utilize fax or auto-fax to deliver pathology reports as a method of report delivery. If you do utilize faxing, which service? (e.g. direct fax to physician office, auto-fax, fax via pre-set code in the fax machine, etc). Also, how much of your pathology reports are faxed this way? (e.g. All, Only STAT's, etc.). Thank-you for taking the time to respond, Sue O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May Court House, NJ 08210 From mari.ann.mailhiot <@t> leica-microsystems.com Wed Jan 19 11:14:11 2005 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] heart specimens Message-ID: Galina Your fixation sounds fine. You may want to rethink your time in the alcohols and Xylene. Animal tissue by itself is dry. Placing the sample in three 100% alcohols for 45 minutes is really taking the bound water out of the tissue. This will account for the dry crumbly sections. Cut back to two 100% alcohols. You can also cut back on the Xylene. Two stations would be more than enough. Too much time in Xylene will make the tissue brittle. The last thing you may want to consider is the time in each station including paraffin. Change the timing to no more than 20 - 30 minutes in each station. There is an excellent book available through NSH. It has a selection of processing protocols for animal tissue from several different researchers. The title is "Aminal Processing Manual". The editors are Gayle Callis and Diane Sterchi. You will find this book to be very usefull! Best Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com Galina Deyneko To: histonet@lists.utsouthwestern.edu Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] heart specimens western.edu 01/18/2005 04:14 PM Galina Deyneko wrote: Dear colleagues please help! I am performing evaluation of the mouse's heart model of cardiac hypertrophy and infarct, but I ran into some problems with microtome sectioning of paraffin blocks of the hearts. It is first time in my practice and I am upset. After harvesting (without perfusion with BNF) hearts are fixed in 10% BNF (I also tried fixation in Bouin's solution and 4% paraphormaldehyde) for 48 hrs and also the time of fixation was extended to 1 week, dehydrated overnight in 70% Ethanol in Cold Room, dissected in short and long axes, and processed in Shandon Processing Center. Ethanol : 60%- 30min Xylene1- 45min 70%-30 min Xylene2- 45 min 95%-40 min Xylene3- 45 min 95%-40 min Wax1- 45 min 100%-40 min Wax2- 1. hrs 100%-45 min Wax3- 1hrs. 100%-45 min I tried to increase and decrease time of processing with and without vacuum and kept in last wax until next morning. Heart tissues appear very dry; they also crumble during sectioning. Before sectioning I kept blocks on ice tray and I also tried to soak them in soap solution. I wetted them with 1% ammonium hydroxide. For waterbath I use distilled water of 43*C. Tissue sections under objective X2 and X4 look unsatisfactory: muscle fibers are torn in many places, crumbled, with pieces torn off, but the remain tissue section mostly without wrinkles. What is significant is that under higher magnification (X100-X400) the cells and nuclei appear fine and clear, but for the photography and measurement my slides are inadequate. Any advice would be highly appreciated. Sincerely, Galina Deyneko. --------------------------------- Do you Yahoo!? Yahoo! Mail - 250MB free storage. Do more. Manage less. --------------------------------- Do you Yahoo!? Read only the mail you want - Yahoo! Mail SpamGuard. _______________________________________________ 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 ekh9535 <@t> bjc.org Wed Jan 19 11:38:02 2005 From: ekh9535 <@t> bjc.org (Erin Herter) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] LEICA ASP300 Message-ID: Hi again all histonetters..... Making a plea again to anyone (else) who has a Leica ASP300. Have you had problems and if so how were they resolved? Our processor fails routinely in the standard cleaning cycle and also during the routine overnight processing. The service company is good, but we are now out of warranty. (We are working on getting that extended because it has been a real "lemon") Thanks, Erin Herter From RBARNHART <@t> summithealth.org Wed Jan 19 11:44:16 2005 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Faxing of Pathology Reports Message-ID: We use autofax and manual fax of reports we receive from one off site surgery center (we autofax to the doctors and manual fax to the surgery center). Doctors that are part of our organization we print directly to thier office since they are on our network. Anytime we need to fax (anything from our LIS, a word document, excel document, etc) we have a feature to fax directly from our computer I believe it is call CDS Desktop Fax. >>> "O'Brien, Sue" 1/19/2005 12:10:08 PM >>> I am looking into how many institutions utilize fax or auto-fax to deliver pathology reports as a method of report delivery. If you do utilize faxing, which service? (e.g. direct fax to physician office, auto-fax, fax via pre-set code in the fax machine, etc). Also, how much of your pathology reports are faxed this way? (e.g. All, Only STAT's, etc.). Thank-you for taking the time to respond, Sue O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May Court House, NJ 08210 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From komo <@t> bu.edu Wed Jan 19 12:31:11 2005 From: komo <@t> bu.edu (Robert Komorowski) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] De-Ionized vs. Distilled Water In-Reply-To: <200501191802.j0JI2kUb028664@relay8.bu.edu> Message-ID: Wonder if some people could provide some useful advice. Are any differences in running immuno using de-ionizied vs. distilled water to do c-fos IEG labelling? Thanks Rob Komorowski From marytedo <@t> hotmail.com Wed Jan 19 13:06:52 2005 From: marytedo <@t> hotmail.com (María Teresa Domínguez) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] contamination/molds In-Reply-To: Message-ID: I usually clean the molds in three steps: I wrappe all the metal molds with a paper towell and put in the oven at 42şC (over night), when the paper is wet with the paraffin, I pass all the molds to a bowl filled of Xilene, and lte them there for 1 hour. To finish all the molds can be dry into the oven at 37şC.The paraffin "rest in peace" in the paper towell, and the Xilene. Good luck! Ht. Maria Teresa Dominguez Anatomic Patology Service Río Grande's Regional Hospital Río Grande, Tierra del Fuego, Argentina. Ma. Teresa >From: "Gudrun Lang" >Reply-To: gu.lang@gmx.at >To: "Histonetliste (Histonetliste)" >Subject: [Histonet] contamination/molds >Date: Wed, 19 Jan 2005 12:52:28 +0100 > > > >-----Ursprüngliche Nachricht----- >Von: Gudrun Lang [mailto:gu.lang@gmx.at] >Gesendet: Mittwoch, 19. Jänner 2005 12:50 >An: 'Molinari, Betsy' >Betreff: AW: [Histonet] contamination/molds > >We clean our molds daily in a ultrasonic bath. Water with tensid, 35 degree; >Paraffin swims on the surface, molds become clean. > >Gudrun Lang >Austria > >-----Ursprüngliche Nachricht----- >Von: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Molinari, >Betsy >Gesendet: Mittwoch, 19. Jänner 2005 12:42 >An: histonet@lists.utsouthwestern.edu >Betreff: [Histonet] contamination/molds > >While on this subject I was just curious..how often do you clean your >molds and how do you clean them? >Betsy Molinari HT(ASCP) >Texas Heart Institute >Cardiovascular Pathology >Houston,TX 77030 >832-355-6524 >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Express yourself instantly with MSN Messenger! [1]MSN Messenger Download today it's FREE! References 1. http://g.msn.com/8HMBEN/2728??PS=47575 From lfidgen <@t> vt.edu Wed Jan 19 13:18:48 2005 From: lfidgen <@t> vt.edu (Laura Fidgen) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] Cleaning molds Message-ID: <6.0.0.22.0.20050119141806.02576958@pop.vt.edu> We put our molds through the cleaning cycle of the tissue processor. Laura L. Fidgen, MScF, BSc, HT(ASCP) Laboratory and Research Practioner VMRCVM, Histopatholgy/Clinical Pathology Dept. Blacksburg, VA 24060-0443 From juan.gutierrez <@t> christushealth.org Wed Jan 19 13:20:27 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] contamination/molds Message-ID: We put them in a big metal bowl with deionized water and regular lab soap and boil them for a while. Let the water cool off and the paraffin solidifies on the surface of the water. Rinse thoroughly and either air dry or put them in the oven. It's always a good idea to stir the molds while they're boiling to release any air or paraffin trapped under molds that are upside down. Good luck. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Molinari, Betsy Sent: Wednesday, January 19, 2005 5:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] contamination/molds While on this subject I was just curious..how often do you clean your molds and how do you clean them? Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology Houston,TX 77030 832-355-6524 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SBarnes <@t> elch.org Wed Jan 19 14:01:07 2005 From: SBarnes <@t> elch.org (Sue Barnes) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] contamination/molds Message-ID: We place the molds on the embedding chamber lid until they warm, so that the moisture doesn't get in the xylene, We place them in a container of xylene for the day, In the afternoon we place the molds in a basin of Dawn dish liquid and hot water, this displaces the xylene. Rinse the molds in clear water and leave on the counter to dry. It has worked for years. From jbirkner <@t> colabserv.com Wed Jan 19 13:58:55 2005 From: jbirkner <@t> colabserv.com (Jeff Birkner) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] workloads Message-ID: I am looking for U.S. Histology lab data. Specifically, what is the average size histology lab, what is the average number of paraffin blocks processed per day, etc. I am also curious about the Laboratory consumables market. Who are the big players? What is the sales volume or market share for reagents (alcohol, xylene, formalin) . Thank you for your assistance. Jeff Birkner, CT(ASCP) Pathology Section Manager Collaborative Laboratory Services, LLC From mprice26 <@t> juno.com Wed Jan 19 14:36:00 2005 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:24:30 2005 Subject: [Histonet] RE: BELAIR INSTRUMENT COMPANY'S # Message-ID: <20050119.123604.12711.236379@webmail32.nyc.untd.com> Histonetters, I need the telephone number to BELAIR INSTRUMENT COMPANY. Thank you. Marsha Price From galinadeyneko <@t> yahoo.com Wed Jan 19 15:26:13 2005 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] heart specimens. Message-ID: <20050119212613.94678.qmail@web14526.mail.yahoo.com> Dear colleagues, I more than appreciate your help and always read histonet with interest. I will follow your advices for next processing. I have new model Shandon Excelsior processor ,.fully automatic and enclosed and I am not able to add or remove any steps, do not have access to reagents and can not use other reagents, only can play with time and on/off vacuum.Thank you again.If you have some more tips I will be glad. I will let you know how the corrections will work. Galina Deyneko Novartis Cambridge, MA 617-871-7613 --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From cfavara <@t> niaid.nih.gov Wed Jan 19 15:32:26 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Formalin vapor fixation Message-ID: Has anyone ever fixed frozen sections with formaldehyde vapor? I have looked in Bancroft and Stevens and they mention it as a suitable fixation. I can not find a protocol. My intention is to place a cotton ball soaked with 0.5ml of NBF in the bottom of a coplin jar, put slides in cover a leave @RT for 15 minutes. Sound good? C Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From Barry.R.Rittman <@t> uth.tmc.edu Wed Jan 19 15:48:35 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry) Date: Fri Sep 16 15:24:31 2005 Subject: [SPAM] - [Histonet] Formalin vapor fixation - Found word(s) list error in the Text body Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0012C9167@UTHEVS3.mail.uthouston.edu> Cynthia Cynthia Vapor fixation is an excellent way to fix frozen sections and thin slices of tissue as the vapor dissolves in the tissue fluid and the problem of loss of materials in solution is minimized. Can use this method ith formalin, osmium tetroxide, alcohol etc. The concentration of the formalin is not that important as long as there is sufficient vapor. The method you suggest should work fine. Don't need to worry about the amount of fluid as long as there is sufficient to produce a good amount of vapor and the tissue does not contact the fluid. I would place the soaked cotton in the container, place a lid on it for 10 minutes or so to give time for vapor to be generated and then add the sections. Of course the type (degree)of fixation as regards bond etc. that you achieve still depends on the amount of time that you leave the tissue exposed. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Favara, Cynthia (NIH/NIAID) Sent: Wednesday, January 19, 2005 3:32 PM To: Histonet Subject: [SPAM] - [Histonet] Formalin vapor fixation - Found word(s) list error in the Text body Has anyone ever fixed frozen sections with formaldehyde vapor? I have looked in Bancroft and Stevens and they mention it as a suitable fixation. I can not find a protocol. My intention is to place a cotton ball soaked with 0.5ml of NBF in the bottom of a coplin jar, put slides in cover a leave @RT for 15 minutes. Sound good? C Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kristenhinkle <@t> yahoo.com Wed Jan 19 16:06:14 2005 From: kristenhinkle <@t> yahoo.com (Kristen Reynolds) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Perfusion suggestions Message-ID: <20050119220615.55980.qmail@web50006.mail.yahoo.com> I need advice on perfusion solutions for rat/monkey brains. I need to use the tissue for both light microscopy and EM. Any suggestions? Kristen Reynolds UT Austin __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From daniel.eberhard <@t> uni-bielefeld.de Wed Jan 19 16:04:08 2005 From: daniel.eberhard <@t> uni-bielefeld.de (Daniel Eberhard) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Formalin vapor fixation In-Reply-To: References: Message-ID: <41EED958.2070302@uni-bielefeld.de> Cynthia, perhaps - http://www.jhc.org/cgi/content/abstract/51/3/401 - might help. Best, Daniel Favara, Cynthia (NIH/NIAID) wrote: >Has anyone ever fixed frozen sections with formaldehyde vapor? I have looked >in Bancroft and Stevens and they mention it as a suitable fixation. I can >not find a protocol. My intention is to place a cotton ball soaked with >0.5ml of NBF in the bottom of a coplin jar, put slides in cover a leave @RT >for 15 minutes. > >Sound good? > >C > >Cynthia Favara >NIAID/NIH/RML/LPVD >903 South 4th Street >Hamilton, MT 59840 >406-363-9317 > >Disclaimer: >The information in this e-mail and any of its attachments is confidential >and may contain sensitive information. It should not be used by anyone who >is not the original intended recipient. If you have received this e-mail in >error please inform the sender and delete it from your mailbox or any other >storage devices. National Institute of Allergy and Infectious Diseases shall >not accept liability for any statements made that are sender's own and not >expressly made on behalf of the NIAID by one of its representatives > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- (-)-(-) --------------------------- \"/ --- Dr. Daniel Eberhard =V= Developmental Biology & Molecular Pathology Graduate Programe on Pattern Formation University of Bielefeld D 33501 Bielefeld/Germany FAX: xx49(0)521-106-5654 (-)-(-) --------------------------- \"/ --- =V= From cfockler <@t> mail1.vcu.edu Wed Jan 19 16:39:10 2005 From: cfockler <@t> mail1.vcu.edu (cfockler@mail1.vcu.edu) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] heart specimens. In-Reply-To: <20050119212613.94678.qmail@web14526.mail.yahoo.com> Message-ID: <200501192239.j0JMdAf10171@caladan.vcu.edu> ------------------- > Dear colleagues, > I more than appreciate your help and always read histonet with interest. I will follow your advices for next processing. > I have new model Shandon Excelsior processor ,.fully automatic and enclosed and I am not able to add or remove any steps, do not have access to reagents and can not use other reagents, only can play with time and on/off vacuum.Thank you again.If you have some more tips I will be glad. I will let you know how the corrections will work. > Galina Deyneko > Novartis Cambridge, MA > 617-871-7613 > > > > --------------------------------- > Do you Yahoo!? > Yahoo! Search presents - Jib Jab's 'Second Term' > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > We have the excelsior as a demo, and if you need to you can access the reagents. You just have to do a dump on them. For us, I can go into the "Check wax, reagent" menu (its the last one on our machine) and you can inspect each reagent and either put it back, or discard it into an empty waste container and replace it with fresh. In order to change the settings you would have to call shandon themselves. I can change ours, but like I said its a demo and if you have an access code in place you'll have to call about it. If you really do want to change the reagents I would just give shandon a call, they are the most helpful, service oriented company I have dealt with. Good luck. Candyce Fockler Histotechnician Anatomic Pathology From cfavara <@t> niaid.nih.gov Wed Jan 19 17:45:34 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Formalin vapor thanks Message-ID: Wonderful replies the most useful so far is this reference for anyone else interested. Has implications for 2 projects I am involved with. Thanks so much to all C http://www.jhc.org/cgi/content/abstract/51/3/401 - Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From cfavara <@t> niaid.nih.gov Wed Jan 19 17:52:16 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Perfusion suggestions Message-ID: Have done some mice with a glut/formaldehyde mix. Do not know the percentages off the top of my head. Following the perfusion we grossly section the brain either coronally or sagittally usually to a 2-4 mm thickness and them do a punch in the area of interest for EM. The remaining specimen is post fixed for light microscopy in NBF for minimum of 24 hours up to 5 days. I believe the EM crew post fixes there specimens as well. Nice thing is that EM seems to be happy and we know exactly where the specimen was taken because the punch is visible on the light microscopy slide. c -----Original Message----- From: Kristen Reynolds [mailto:kristenhinkle@yahoo.com] Sent: Wednesday, January 19, 2005 3:06 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Perfusion suggestions I need advice on perfusion solutions for rat/monkey brains. I need to use the tissue for both light microscopy and EM. Any suggestions? Kristen Reynolds UT Austin __________________________________________________ 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 cindipqr <@t> wowway.com Wed Jan 19 18:12:01 2005 From: cindipqr <@t> wowway.com (cindipqr@wowway.com) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] PLP fixative Message-ID: <20050120000855.M49506@wowway.com> Hi, does anyone have the recipe for PLP fixative and how it is used? After fixation can the tissue (brain) be processed thru paraffin as usual? Cindi Roberts Neurology Research Henry Ford Hospital -- WOW! Homepage (http://www.wowway.com) From FANDORFP <@t> aol.com Wed Jan 19 18:30:46 2005 From: FANDORFP <@t> aol.com (FANDORFP@aol.com) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Re: Unsubscribe Message-ID: <6A3509AD.3E89F693.00678004@aol.com> Please Unsubscibe. thank-you. From RCHIOVETTI <@t> aol.com Wed Jan 19 18:41:32 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] PLP fixative Message-ID: <128.54808e9e.2f20583c@aol.com> In a message dated 1/19/2005 5:12:42 PM US Mountain Standard Time, cindipqr@wowway.com writes: > does anyone have the recipe for PLP fixative and how it is used? > After fixation can the tissue (brain) be processed thru paraffin as usual? > Cindi Roberts > Neurology Research > Henry Ford Hospital > > Cindi, The recipe for PLP (Periodate-Lysine-Paraformaldehyde) fixative and a sample protocol (at least for immuno) using perfusion fixation is on the Chemicon website at: http://www.chemicon.com/techsupp/Protocol/perfusion.asp There are a few instances of using PLP with paraffin embedding, so the processing should work OK. Whether you can retrieve reactivity from paraffin sections seems to be related to the antigen. For example, see the NIEHS website: http://dir.niehs.nih.gov/dirlep/immuno/protocols.htm Cheers, Bob Chiovetti From phesch <@t> charter.net Wed Jan 19 19:39:30 2005 From: phesch <@t> charter.net (Pam Hesch) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] LEICA ASP300 References: Message-ID: <002301c4fe90$e3169430$72d0c518@yourae066c3a9b> Erin, Where I worked I had 4 ASP300, there were some problems in the beginning but they are a workhorse. If you would like other information feel free to give me a call. (507)289-4046 Pam ----- Original Message ----- From: "Erin Herter" To: Sent: Wednesday, January 19, 2005 11:38 AM Subject: [Histonet] LEICA ASP300 > Hi again all histonetters..... > Making a plea again to anyone (else) who has a Leica ASP300. Have you > had problems and if so how were they resolved? Our processor fails > routinely in the standard cleaning cycle and also during the routine > overnight processing. The service company is good, but we are now out > of warranty. (We are working on getting that extended because it has > been a real "lemon") > Thanks, > Erin Herter > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Thu Jan 20 00:35:10 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Perfusion suggestions References: <20050119220615.55980.qmail@web50006.mail.yahoo.com> Message-ID: <41EF511E.9862D624@uwo.ca> Is there nobody near you who understands the ins and outs of perfusion fixation? What about your boss? It isn't possible to give complete instructions in an email reply. The answer to your "Any suggestions?" has to be: Get your boss to send you for a course on EM techniques for research, and get her (or him) to buy a few books for the lab. Your boss must have a large research grant to be doing EM work with monkey tissues. If he/she cannot answer your simple question, why doesn't she/he go to the library, or send you there with clear instructions? How did your boss get a grant to finance EM of monkey tissues without having a clue about how to do the work? Forward this reply to your mistress/master and advise him/her/it to: (a) go to the library (b) buy a few books for the lab (c) read in library and from books (d) become a Histonet subscriber (e) all of the above (f) invoke Harry Potter baddy's curse upon me. ---- John Kiernan London, Canada. _______________________________________________ Kristen Reynolds wrote: > > I need advice on perfusion solutions for rat/monkey > brains. I need to use the tissue for both light > microscopy and EM. Any suggestions? > > Kristen Reynolds > UT Austin ________________________________________________ From Kemlo.Rogerson <@t> elht.nhs.uk Thu Jan 20 02:06:48 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Faxing of Pathology Reports[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EE94@bhrv-nt-11.bhrv.nwest.nhs.uk> Maybe it's wise to make sure that the FAX you are sending goes to a secure machine by whatever means. Cleaners and others may take interest in a result on their mate(s); get a disclaimer from the recipient accepting responsibility for the secureness of their telecommunication system and file it. Plus I think you have a responsibility to send to the correct FAX and to record that. -----Original Message----- From: O'Brien, Sue [mailto:histo@bthosp.com] Sent: 19 January 2005 17:10 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Faxing of Pathology Reports[Scanned] I am looking into how many institutions utilize fax or auto-fax to deliver pathology reports as a method of report delivery. If you do utilize faxing, which service? (e.g. direct fax to physician office, auto-fax, fax via pre-set code in the fax machine, etc). Also, how much of your pathology reports are faxed this way? (e.g. All, Only STAT's, etc.). Thank-you for taking the time to respond, Sue O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May Court House, NJ 08210 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hasselblatt <@t> uni-muenster.de Thu Jan 20 03:46:14 2005 From: hasselblatt <@t> uni-muenster.de (Martin Hasselblatt) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Collagen III antibody Message-ID: <41EF7DE6.3080601@uni-muenster.de> Hi, is anybody aware of an type III Collagen antibody that works on formalin-fixed,paraffin-embedded tissue? I sure hope that someone out there has some experience to share on this subject. Thanks in advance! Martin -- __________________________________________________________________________ Martin Hasselblatt MD Institute of Neuropathology Universit?tsklinikum M?nster Domagkstr. 19 48129 M?nster, Germany http://www.klinikum.uni-muenster.de/institute/npatho/mitarbeiter/hasselblatt/index.html From antje.marcantonio <@t> pharma.novartis.com Thu Jan 20 04:15:50 2005 From: antje.marcantonio <@t> pharma.novartis.com (antje.marcantonio@pharma.novartis.com) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Collagen III antibody Message-ID: Hi Martin we used the mAb to Collagen III from BioGenex,clone HWD1.1 for FFPE tissue. Absolutely necessary: pretreatment with pepsin as indicated on the datasheet. We used it in monkeys and dogs with a concentration of 1:50, incubation in the fridge over night. Hope this helps. Antje Marcantonio Novartis Pharma AG BU Transplantation Research Basel, Switzerland Tel. +41 61 324 6730 antje.marcantonio@pharma.novartis.com From Terry.Marshall <@t> rothgen.nhs.uk Thu Jan 20 04:54:21 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Faxing of Pathology Reports[Scanned] Message-ID: As you may gather from Kemlo's post, here in the UK we are rather hot on patient confidentiality. This extends to protecting patient's path reports from being seen by most of the world, which would have no interest or understanding of them whatsoever. Be that as it may vast amounts of money and mental energy are expended in this aim. On the rare occasions we fax, our practise is to be on the phone during a fax, so that its safe receipt by the requester can be recorded, or, if not feasible, be on the phone immediately before and after for the same purpose. BTW - I acknowledge that this aspect of faxing was not part of the original query. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Kemlo Rogerson [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 20 January 2005 08:07 To: 'O'Brien, Sue'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Faxing of Pathology Reports[Scanned] Maybe it's wise to make sure that the FAX you are sending goes to a secure machine by whatever means. Cleaners and others may take interest in a result on their mate(s); get a disclaimer from the recipient accepting responsibility for the secureness of their telecommunication system and file it. Plus I think you have a responsibility to send to the correct FAX and to record that. -----Original Message----- From: O'Brien, Sue [mailto:histo@bthosp.com] Sent: 19 January 2005 17:10 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Faxing of Pathology Reports[Scanned] I am looking into how many institutions utilize fax or auto-fax to deliver pathology reports as a method of report delivery. If you do utilize faxing, which service? (e.g. direct fax to physician office, auto-fax, fax via pre-set code in the fax machine, etc). Also, how much of your pathology reports are faxed this way? (e.g. All, Only STAT's, etc.). Thank-you for taking the time to respond, Sue O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May Court House, NJ 08210 _______________________________________________ Histonet mailing list Histonet@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 Don.Birgerson <@t> leica-microsystems.com Thu Jan 20 07:17:35 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] RE: BELAIR INSTRUMENT COMPANY'S # Message-ID: Hi Marsha, Belair Instruments at 1-800-783-9424 or 1-973-912-8900 ask for Sheila and she can direct you. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "mprice26@juno.com" whnt.nhs.uk Thu Jan 20 08:46:23 2005 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Re: heart specimens - Shandon processing Message-ID: Tina You don't say which Shandon processor you were using. We have 2, a Pathcentre and an Excelsior. There were problems with the Excelsior at the beginning but Shandon (UK) were brilliant in looking after us during this time. I think you must have had an undiagnosed problem with yours as merely changing processor if all else stays the same should make little difference. By the way when looking to buy the Pathcentre I conacted Bayer for a quote etc, the local rep couldn't even be bothered to telephone to follow up on the quote. Not the service I expected as a previous Bayer user in another hospital. 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 From: bettina.hutz@orionpharma.com Subject: RE: [Histonet] heart specimens To: galinadeyneko@yahoo.com, histonet@lists.utsouthwestern.edu Message-ID: <243E1A79AF5D89438C70513DBADAA4F0019E40A1@sfies-exchange1.orionnet.org> Content-Type: text/plain; charset="iso-8859-1" Hello, I am working on a similar heart model, and first of all: I replaced the Shandon Processing Center by Sakura Tissue Tek VIP5, and suddenly everything works fine:) Nothing against Shandon, but their Histology equipment sucks a big time! Greetings, Tina From mcauliff <@t> umdnj.edu Thu Jan 20 12:00:08 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Perfusion suggestions In-Reply-To: <20050119220615.55980.qmail@web50006.mail.yahoo.com> References: <20050119220615.55980.qmail@web50006.mail.yahoo.com> Message-ID: <41EFF1A8.5080701@umdnj.edu> Dear Kristen: I will second John Kiernan's response, suggestions in an e-mail won't do the job. Find a lab that does perfusions for EM and spend a day or two there to observe their materials and methods. That way you can do it right the first time. Geoff Kristen Reynolds wrote: >I need advice on perfusion solutions for rat/monkey >brains. I need to use the tissue for both light >microscopy and EM. Any suggestions? > >Kristen Reynolds >UT Austin > >__________________________________________________ >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 > > > -- -- ********************************************** 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 gcallis <@t> montana.edu Thu Jan 20 09:31:28 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] PLP fixative In-Reply-To: <128.54808e9e.2f20583c@aol.com> References: <128.54808e9e.2f20583c@aol.com> Message-ID: <6.0.0.22.1.20050120081726.01b505d0@gemini.msu.montana.edu> Our prion people do this routinely . The PLP is made up fresh just before animal euthanasia begins. PLP perfusion of rodent model via heart (after anesthetic) using a tidy little circulating pump followed by 5 hours immersion fixation for tissues dissected out post-perfusion. Each brain (mouse, hamster, etc) has a different processing schedule protocol to avoid overdehydration. Brain and spinal cord, other tissues section without problems as long as hair and any bone fragments are avoided at dissection. To facilitate tongue fixation, it is bisected before immersion. Brains are sliced using a matrix (go to www.myneurolab.com, hope this is correct) for precise sectioning, orientation of brain). IHC for PrP antigens is done with sections are put into formic acid, a form of retrieval for this unique PrP. They have excellent results with IHC. PLP fixed tissues are never processed with or after NBF fixed tissue. The researcher is concerned NBF (even a sniff of!) will compromise antigen. Frequently, PLP perfused hamster brain and tongue are cryoprotected with 30% sucrose and cut at 50 um with dedicated Leica 1850 cryostat using antiroll device. >Cindi, > >The recipe for PLP (Periodate-Lysine-Paraformaldehyde) fixative and a sample >protocol (at least for immuno) using perfusion fixation is on the Chemicon >website at: >http://www.chemicon.com/techsupp/Protocol/perfusion.asp > >There are a few instances of using PLP with paraffin embedding, so the >processing should work OK. Whether you can retrieve reactivity from paraffin >sections seems to be related to the antigen. For example, see the NIEHS >website: >http://dir.niehs.nih.gov/dirlep/immuno/protocols.htm > >Cheers, > >Bob Chiovetti > >_______________________________________________ >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 kristenhinkle <@t> yahoo.com Thu Jan 20 09:47:16 2005 From: kristenhinkle <@t> yahoo.com (Kristen Reynolds) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] perfusion suggestions Message-ID: <20050120154716.70554.qmail@web50010.mail.yahoo.com> Wow, I can't believe how rude of a response I got. I thought this was for helpful comments. I know how to perfuse for light microscopy and EM. I just thought the fixative combination for wanting to do both on the same tissue might be out there. I guess I won't be asking histonet anything anymore. __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From anaflorenti <@t> yahoo.com Thu Jan 20 10:31:18 2005 From: anaflorenti <@t> yahoo.com (Anne Flare) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Zink fixation of frozen tissue sections protocol?! Message-ID: <20050120163119.53195.qmail@web52606.mail.yahoo.com> Hello everybody! Does anyone have some protocols for Zink fixation of frozen tissue sections? I would really appreciate any help! Thank you! __________________________________ Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. http://promotions.yahoo.com/new_mail From mcauliff <@t> umdnj.edu Thu Jan 20 13:44:27 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] perfusion suggestions follow up In-Reply-To: <20050120154716.70554.qmail@web50010.mail.yahoo.com> References: <20050120154716.70554.qmail@web50010.mail.yahoo.com> Message-ID: <41F00A1B.6060101@umdnj.edu> Dear Kristen: Your original posting (see below) gave no indication that you were familiar with perfusion for LM or EM. You did not ask for fixative composition, you just asked for "any suggestions". Given the broad scope of your question, the advice you got WAS helpful. The fixative combination for wanting to do both on the same tissue IS out there, but you have to be more specific in your question. The correct fixative combinatio will depend on: 1. Are you doing histochemistry? If so, what reaction? Pre-embedding or post-embedding reaction? 2. Are you doing immunohistochemistry? If so, what antigen? Pre-embedding or post-embedding reaction? 3. What plastic are you embedding in? 4. What routine stains do you need to do? 5. What part of the brain will you be looking at? The brain is a big place. 6. Do you need to do LM and EM on the same piece of tissue/same cells? Advice on HistoNet is free. Some of it is quite good. Checking published papers in refereed journals and looking at the results is always a good idea, espcially given the cost of experimental animals. Geoff Kristen Reynolds wrote: I need advice on perfusion solutions for rat/monkey brains. I need to use the tissue for both light microscopy and EM. Any suggestions? >Wow, I can't believe how rude of a response I got. I >thought this was for helpful comments. I know how to >perfuse for light microscopy and EM. I just thought >the fixative combination for wanting to do both on the >same tissue might be out there. I guess I won't be >asking histonet anything anymore. > >__________________________________________________ >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 > > > -- -- ********************************************** 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 julien_lambreydesouza <@t> uqar.qc.ca Thu Jan 20 11:07:15 2005 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] lab management software Message-ID: <014e01c4ff12$821f4c80$9310d784@uqar.qc.ca> Hello all, Does anybody know of a good lab-management software (free if possible) that would help out in inventory keeping, product specifications, etc. Thanks, Julien Lambrey de Souza Biologie ?volutive, Universit? du Qu?bec ? Rimouski, (418) 723-1986 #1438 From jkiernan <@t> uwo.ca Thu Jan 20 11:24:28 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Formalin vapor fixation References: <41EED958.2070302@uni-bielefeld.de> Message-ID: <41EFE94C.76487A84@uwo.ca> Daniel's 2003 paper in J Histochem Cytochem has excellent coloured photomicrographs that show what can be achieved by vapour fixation of cryosections. It's worth looking up the full text as well as the abstract. John Kiernan London, Canada ------------------------------------------------ Daniel Eberhard wrote: > > Cynthia, > perhaps - http://www.jhc.org/cgi/content/abstract/51/3/401 > - might help. > Best, > Daniel > > Favara, Cynthia (NIH/NIAID) wrote: > > >Has anyone ever fixed frozen sections with formaldehyde vapor? I have looked > >in Bancroft and Stevens and they mention it as a suitable fixation. I can > >not find a protocol. My intention is to place a cotton ball soaked with > >0.5ml of NBF in the bottom of a coplin jar, put slides in cover a leave @RT > >for 15 minutes. > > > >Sound good? > > > >C > > > >Cynthia Favara > >NIAID/NIH/RML/LPVD > >903 South 4th Street > >Hamilton, MT 59840 > >406-363-9317 > > > >Disclaimer: > >The information in this e-mail and any of its attachments is confidential > >and may contain sensitive information. It should not be used by anyone who > >is not the original intended recipient. If you have received this e-mail in > >error please inform the sender and delete it from your mailbox or any other > >storage devices. National Institute of Allergy and Infectious Diseases shall > >not accept liability for any statements made that are sender's own and not > >expressly made on behalf of the NIAID by one of its representatives > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > -- > (-)-(-) > --------------------------- \"/ --- > Dr. Daniel Eberhard =V= > > Developmental Biology > & Molecular Pathology > > Graduate Programe on Pattern Formation > > University of Bielefeld > D 33501 Bielefeld/Germany > > FAX: xx49(0)521-106-5654 (-)-(-) > --------------------------- \"/ --- > =V= > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Thu Jan 20 11:59:53 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Monastral blue References: <8D6F233E2A5D574B929F3944F3316FD01A58EA@EXCH2K3.utmb.edu> Message-ID: <41EFF199.1DD54B5A@uwo.ca> Monastral blue B is not a dye; it's a pigment (CI 74160; Pigment blue 15). Its chemical name is copper(II) phthalocyanine, and this is in the Sigma and Aldrich catalogues. Aldrich sells three grades, with a wide variation in prices. My guess is that the cheapest would be OK. None are hugely expensive, however, so it might be best to try more than one. For vascular permeability studies you need a small particle size that will remain in suspension. Here are a few references that may help. The first one in the list shows that the circulation time is only 3-4 minutes. That is similar to Indian ink, which you might be able to use instead. Drake WT, Creighan M, Sims DE (1992) Limitations of monastral blue as a vascular label: rapid rate of clearance is age-dependent, and interactions with anesthetics depress arterial blood pressure in rats. Microscopy Research and Technique 23: 219-224. Dux M, Jancso G (1994) A new technique for the direct demonstration of overlapping cutaneous innervation territories of peptidergic C-fibre afferents of rat hindlimb nerves. Journal of Neuroscience Methods 55: 47-52. Baluk P, Hirata A, Thurston G, Fujiwara T, Neal CR, Michel CC, McDonald DM (1997) Endothelial gaps: Time course of formation and closure in inflamed venules of rats. American Journal of Physiology - Lung Cellular and Molecular Physiology 16: L155-L170. Hu P, Pollard J, Hunt N, Taylor J, Chanling T (1998) Microvascular and cellular responses in the optic nerve of rats with acute experimental allergic encephalomyelitis (EAE). Brain Pathology 8: 475-486. Horobin, RW (2002) Phthalocyanines, porphyrins and related aza[18]annulenes. Chapter 26 in Conn's Biological Stains, 10th ed. Oxford: BIOS. (Contains basic information, and refers to recent literature of various uses of monastral blue B) -- ------------------------------- 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 _______________________________ "Hawkins, Hal K." wrote: > > > We need to study changes in vascular permeability in animal models using > Monastral Blue B dye. Apparently Sigma-Aldrich no longer supplies this > pigment. Does anyone know of another source, or would anyone be willing > to share/sell us some? Alternatively, is there another colloid that is > useful in the same way at the level of both light and electron > microscopy? > > Thanks, > Hal Hawkins > University of Texas Medical Branch > Galveston, Texas > hhawkins@utmb.edu > From subratab <@t> bdonline.com Thu Jan 20 12:02:21 2005 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] NIH image J surface area unit Message-ID: <200501201804.j0KI4YYb008313@mailout.proshikanet.com> Dear Histo experts My friend is measuring glomerular surface area by using Image J software. He is in trouble to get the unit of the area calculated by the software, I mean, if the calculated area is to be expressed in square milimeter or square micrometer. He is capturing the figures from microscope with X100 magnification (oil-emersion lense) and then analyzing the glomerular surface area by the software in a computer. Could anybody please help to solve this. Sincerely Subrata Biswas University of Campinas SP, Brazil. -- http://www.histosearch.com/homepages/DrSubrataKumarBiswas.html From flemons <@t> bhset.org Thu Jan 20 12:26:57 2005 From: flemons <@t> bhset.org (Fran Lemons) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Certification requirements Message-ID: Has anyone out there seen anything in writing about not being able to work in histology labs without HT certification? Thanks Fran Walker From pruegg <@t> ihctech.net Thu Jan 20 13:16:05 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] frozens on fat to image GFP Message-ID: <200501201916.j0KJG02p023804@chip.viawest.net> Help! Here is the situation. An investigator wants me to cut fat tissue (briefly paraformaldehyde fixed,snap frozen in OCT) which does not cut well, so I had the brillant (I thought) idea of using the Instrumedics tape transfer system. I cut the sections and transfered them to the coated slides, air dryed them, and they looked pretty good. Since they want to preserve the fat I thought I would just use some aqueous mount and put a cs on them so they could look at the sections under UV. Well there was some kind of reaction from the ?water or glycerine or something that turned white so we can't see the tissue. Tryed soaking the slides in water to remove mount, the cs came off put the white reaction did not. so is the water reacting with the polymer coating (?might be GMA). Should I take these sections thru solvent anyway to get them coverslipped so they are clear??? Patsy From jhabecke <@t> seattlecca.org Thu Jan 20 13:27:09 2005 From: jhabecke <@t> seattlecca.org (Randolph-Habecker, Julie) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Rudeness on histonet Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAE7E@wala01.seattlecca.org> Dear Kristen and others: I too feel that Kristen was treated rudely. I have noticed this problem from time to time on HistoNet and I feel I have to say something. Some people generally assume that because you ask an open question that you are clueless. While you might get better answers by being more specific, the better way to respond to "any suggestions" is to ask if you could provide more details. Just assuming that you and your boss have no idea what you are doing and have ventured into this project naively is not fair. That response was not helpful - it was nothing but rude. Did they suggest a class or a book that they found practically good? No! The response was condescending and pointless. Please ask for clarification before you humiliate a person. And please do not tell someone they deserved to be treated rudely - that is even worse! People write into histonet with questions because there is a wealth of knowledge amongst the subscribers. This is valuable information and constitutes years and years of experience. Keep in mind, all of our experiences have been different. Just because a person might not have experience in one area doesn't mean that they are not very knowledgeable in another. In other words, if you are rude to a person they might not want to share valuable information with you when you ask. Or worse, they might leave the Histonet and deprive all of us of their knowledge! I too asked an open question one time about processing tissue with plastic beads in it and received an incredibly condescending response instructing me on how to design an experiment. I have a Ph.D. for god's sake. I have spent 15 years designing experiments. I also saw a young student publicly filleted for cheating when her instructor encouraged her to send her inquiry out to the Histonet. Kristen, please don't leave the histonet because of one (incredibly) rude response. Take it for what it is - useless! Please hold out for helpful people that actually have information to contribute. Hang in there! 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 Message: 2 Date: Thu, 20 Jan 2005 11:44:27 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] perfusion suggestions follow up To: Kristen Reynolds Cc: histonet@lists.utsouthwestern.edu Message-ID: <41F00A1B.6060101@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Dear Kristen: Your original posting (see below) gave no indication that you were familiar with perfusion for LM or EM. You did not ask for fixative composition, you just asked for "any suggestions". Given the broad scope of your question, the advice you got WAS helpful. The fixative combination for wanting to do both on the same tissue IS out there, but you have to be more specific in your question. The correct fixative combinatio will depend on: 1. Are you doing histochemistry? If so, what reaction? Pre-embedding or post-embedding reaction? 2. Are you doing immunohistochemistry? If so, what antigen? Pre-embedding or post-embedding reaction? 3. What plastic are you embedding in? 4. What routine stains do you need to do? 5. What part of the brain will you be looking at? The brain is a big place. 6. Do you need to do LM and EM on the same piece of tissue/same cells? Advice on HistoNet is free. Some of it is quite good. Checking published papers in refereed journals and looking at the results is always a good idea, espcially given the cost of experimental animals. Geoff Kristen Reynolds wrote: I need advice on perfusion solutions for rat/monkey brains. I need to use the tissue for both light microscopy and EM. Any suggestions? >Wow, I can't believe how rude of a response I got. I >thought this was for helpful comments. I know how to >perfuse for light microscopy and EM. I just thought >the fixative combination for wanting to do both on the >same tissue might be out there. I guess I won't be >asking histonet anything anymore. > >__________________________________________________ > 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 rocan <@t> mac.com Thu Jan 20 13:28:47 2005 From: rocan <@t> mac.com (Rocan) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] NIH image J surface area unit In-Reply-To: <200501201804.j0KI4YYb008313@mailout.proshikanet.com> References: <200501201804.j0KI4YYb008313@mailout.proshikanet.com> Message-ID: <816F081F-6B19-11D9-8D0A-000A9589219E@mac.com> I guess there are a couple of ways to do this. I use a stage micrometer (you can google "stage micrometer" and find it on the pictures). I take a picture of the ruler imprinted. Typically these rulers have a one millimeter ruler and 100 divisions of 10 micrometers each. So, with the photo using the same scope and objective you go to any of these programs like NIH ImageJ and open the document (same size as the photos you are measuring). There is a command where you place the mouse on one end drag and click on the other end of the markings. So, if you drag it over ten marking you then instruct the program that that distance is 100 micrometers. Then using these settings you do your measurements and the measurements would then be very precise and will be in micrometers. I do this with a different program but I know you can do it with ImageJ. I hope this helps. Rocio ----- Dr.Rocio Sierra-Honigmann Director Engineered Wound Repair Laboratory Cedars Sinai Research Institute Davis 1091 310-423-1882 Honigmannr@cshs.org On Jan 20, 2005, at 10:02 AM, Subratab wrote: > Dear Histo experts > My friend is measuring glomerular surface area by using Image J > software. He > is in trouble to get the unit of the area calculated by the software, I > mean, if the calculated area is to be expressed in square milimeter or > square micrometer. > He is capturing the figures from microscope with X100 magnification > (oil-emersion lense) and then analyzing the glomerular surface area by > the > software in a computer. > Could anybody please help to solve this. > Sincerely > > Subrata Biswas > University of Campinas > SP, Brazil. > > -- > http://www.histosearch.com/homepages/DrSubrataKumarBiswas.html > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mrsgbd2001 <@t> yahoo.com Thu Jan 20 14:09:42 2005 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Certification requirements In-Reply-To: Message-ID: <20050120200942.71181.qmail@web52708.mail.yahoo.com> Fran, I believe it's based on what state you work in. I know that Florida requires an ASCP certification as well as a Florida license. Tennessee, where I work, does not require it. Gareth Davis Fran Lemons wrote: Has anyone out there seen anything in writing about not being able to work in histology labs without HT certification? Thanks Fran Walker _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From mrsgbd2001 <@t> yahoo.com Thu Jan 20 14:14:32 2005 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Rudeness on histonet In-Reply-To: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAE7E@wala01.seattlecca.org> Message-ID: <20050120201432.90980.qmail@web52710.mail.yahoo.com> Here! Here! to Julie. I have also experienced the rudeness and seen others put down and treated as if they were stupid. It's very discouraging. Gareth "Randolph-Habecker, Julie" wrote: Dear Kristen and others: I too feel that Kristen was treated rudely. I have noticed this problem from time to time on HistoNet and I feel I have to say something. Some people generally assume that because you ask an open question that you are clueless. While you might get better answers by being more specific, the better way to respond to "any suggestions" is to ask if you could provide more details. Just assuming that you and your boss have no idea what you are doing and have ventured into this project naively is not fair. That response was not helpful - it was nothing but rude. Did they suggest a class or a book that they found practically good? No! The response was condescending and pointless. Please ask for clarification before you humiliate a person. And please do not tell someone they deserved to be treated rudely - that is even worse! People write into histonet with questions because there is a wealth of knowledge amongst the subscribers. This is valuable information and constitutes years and years of experience. Keep in mind, all of our experiences have been different. Just because a person might not have experience in one area doesn't mean that they are not very knowledgeable in another. In other words, if you are rude to a person they might not want to share valuable information with you when you ask. Or worse, they might leave the Histonet and deprive all of us of their knowledge! I too asked an open question one time about processing tissue with plastic beads in it and received an incredibly condescending response instructing me on how to design an experiment. I have a Ph.D. for god's sake. I have spent 15 years designing experiments. I also saw a young student publicly filleted for cheating when her instructor encouraged her to send her inquiry out to the Histonet. Kristen, please don't leave the histonet because of one (incredibly) rude response. Take it for what it is - useless! Please hold out for helpful people that actually have information to contribute. Hang in there! 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 Message: 2 Date: Thu, 20 Jan 2005 11:44:27 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] perfusion suggestions follow up To: Kristen Reynolds Cc: histonet@lists.utsouthwestern.edu Message-ID: <41F00A1B.6060101@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Dear Kristen: Your original posting (see below) gave no indication that you were familiar with perfusion for LM or EM. You did not ask for fixative composition, you just asked for "any suggestions". Given the broad scope of your question, the advice you got WAS helpful. The fixative combination for wanting to do both on the same tissue IS out there, but you have to be more specific in your question. The correct fixative combinatio will depend on: 1. Are you doing histochemistry? If so, what reaction? Pre-embedding or post-embedding reaction? 2. Are you doing immunohistochemistry? If so, what antigen? Pre-embedding or post-embedding reaction? 3. What plastic are you embedding in? 4. What routine stains do you need to do? 5. What part of the brain will you be looking at? The brain is a big place. 6. Do you need to do LM and EM on the same piece of tissue/same cells? Advice on HistoNet is free. Some of it is quite good. Checking published papers in refereed journals and looking at the results is always a good idea, espcially given the cost of experimental animals. Geoff Kristen Reynolds wrote: I need advice on perfusion solutions for rat/monkey brains. I need to use the tissue for both light microscopy and EM. Any suggestions? >Wow, I can't believe how rude of a response I got. I >thought this was for helpful comments. I know how to >perfuse for light microscopy and EM. I just thought >the fixative combination for wanting to do both on the >same tissue might be out there. I guess I won't be >asking histonet anything anymore. > >__________________________________________________ > 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 --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' From dlcowie <@t> prodigy.net Thu Jan 20 14:49:37 2005 From: dlcowie <@t> prodigy.net (Dawn Cowie) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] certification requirements to work in histo labs Message-ID: <20050120204937.7153.qmail@web81008.mail.yahoo.com> Fran, In Florida where I am, to perform tech duties you must be licensed. However, you may employ non techs to work as lab aides. These aides may perform a wide variety of duties that usually are done by techs. Specifically what they cannot do is: embed, cut, stain, frozen sections etc. What they can do is: change reagents, coverslip, start and stop an automated stainer. I hope this helps. Dawn Cowie, HT Pensacola Path From vazquezr <@t> ohsu.edu Thu Jan 20 14:57:54 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Rudeness on histonet Message-ID: BRAVO ladies for speaking your mind!!!! :0) Robyn >>> "Randolph-Habecker, Julie" 1/20/2005 11:27:09 AM >>> Dear Kristen and others: I too feel that Kristen was treated rudely. I have noticed this problem from time to time on HistoNet and I feel I have to say something. Some people generally assume that because you ask an open question that you are clueless. While you might get better answers by being more specific, the better way to respond to "any suggestions" is to ask if you could provide more details. Just assuming that you and your boss have no idea what you are doing and have ventured into this project naively is not fair. That response was not helpful - it was nothing but rude. Did they suggest a class or a book that they found practically good? No! The response was condescending and pointless. Please ask for clarification before you humiliate a person. And please do not tell someone they deserved to be treated rudely - that is even worse! People write into histonet with questions because there is a wealth of knowledge amongst the subscribers. This is valuable information and constitutes years and years of experience. Keep in mind, all of our experiences have been different. Just because a person might not have experience in one area doesn't mean that they are not very knowledgeable in another. In other words, if you are rude to a person they might not want to share valuable information with you when you ask. Or worse, they might leave the Histonet and deprive all of us of their knowledge! I too asked an open question one time about processing tissue with plastic beads in it and received an incredibly condescending response instructing me on how to design an experiment. I have a Ph.D. for god's sake. I have spent 15 years designing experiments. I also saw a young student publicly filleted for cheating when her instructor encouraged her to send her inquiry out to the Histonet. Kristen, please don't leave the histonet because of one (incredibly) rude response. Take it for what it is - useless! Please hold out for helpful people that actually have information to contribute. Hang in there! 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 Message: 2 Date: Thu, 20 Jan 2005 11:44:27 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] perfusion suggestions follow up To: Kristen Reynolds Cc: histonet@lists.utsouthwestern.edu Message-ID: <41F00A1B.6060101@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Dear Kristen: Your original posting (see below) gave no indication that you were familiar with perfusion for LM or EM. You did not ask for fixative composition, you just asked for "any suggestions". Given the broad scope of your question, the advice you got WAS helpful. The fixative combination for wanting to do both on the same tissue IS out there, but you have to be more specific in your question. The correct fixative combinatio will depend on: 1. Are you doing histochemistry? If so, what reaction? Pre-embedding or post-embedding reaction? 2. Are you doing immunohistochemistry? If so, what antigen? Pre-embedding or post-embedding reaction? 3. What plastic are you embedding in? 4. What routine stains do you need to do? 5. What part of the brain will you be looking at? The brain is a big place. 6. Do you need to do LM and EM on the same piece of tissue/same cells? Advice on HistoNet is free. Some of it is quite good. Checking published papers in refereed journals and looking at the results is always a good idea, espcially given the cost of experimental animals. Geoff Kristen Reynolds wrote: I need advice on perfusion solutions for rat/monkey brains. I need to use the tissue for both light microscopy and EM. Any suggestions? >Wow, I can't believe how rude of a response I got. I >thought this was for helpful comments. I know how to >perfuse for light microscopy and EM. I just thought >the fixative combination for wanting to do both on the >same tissue might be out there. I guess I won't be >asking histonet anything anymore. > >__________________________________________________ > 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 stewart_georgia <@t> sbcglobal.net Thu Jan 20 14:58:14 2005 From: stewart_georgia <@t> sbcglobal.net (Georgia Stewart) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] HIPPA rules and fax transmittals going to wrong person Message-ID: <20050120205814.70166.qmail@web81402.mail.yahoo.com> Hello and Good Afternoon Everyone In this era of HIPPA rules and regulations in the United States, I have not seen any comment on HIPPA as it relates to fax transmittal's of reports that might go to the wrong person/facility. If a report goes to the the wrong person/facility, isn't this is a breach of patient confidentiality covered by HIPPA? How are you handling faxs that go, or might go, to the wrong person/facility? Appreciate your comments. Georgia Stewart, BS, HTL Georgia From Barry.R.Rittman <@t> uth.tmc.edu Thu Jan 20 15:00:39 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] perfusion suggestions Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0012C92C3@UTHEVS3.mail.uthouston.edu> Kristen I am not trying to justify any response that you received but I think that I understand why the response was phrased in that particular manner. Many of the questions that arrive at Histonet are very specific, and generally get a lot of responses. The broader questions are often difficult to answer (and in general I personally do not respond to these) and often receive few responses. It is usually not possible to determine the level of knowledge of the individual who is asking the question and there may be a wide variety of answers. Here on Histonet we are often between a rock and a hard place. I believe that it is a natural tendency nowadays to pose the question in the shortest way possible on the assumption that nobody wishes to spend their time to read a long discourse. This unfortunately often results in a communication that is open to a wide variety of interpretations. One interpretation is that the individual posing the question wants to be informed about all aspects of a particular technique from anesthetizing an animal to interpreting the final result, rather than carrying out the initial groundwork themselves. There is usually no way to judge if this is true from the original communications. This is the era of rapid fire information but unfortunately the recipients do not always have the same background or experiences to view that information in the same light. In many cases I feel that in response to complex questions, individuals with the appropriate expertise should provide their telephone number so that a dialogue can be set up. While Histonet is very useful, a one on one dialogue generally will provide the most relevant information. I hope that you will continue to ask questions on Histonet. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Reynolds Sent: Thursday, January 20, 2005 9:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] perfusion suggestions Wow, I can't believe how rude of a response I got. I thought this was for helpful comments. I know how to perfuse for light microscopy and EM. I just thought the fixative combination for wanting to do both on the same tissue might be out there. I guess I won't be asking histonet anything anymore. __________________________________________________ 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 victor <@t> pathology.washington.edu Thu Jan 20 15:24:08 2005 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] HIPPA rules and fax transmittals going to wrong person In-Reply-To: <20050120205814.70166.qmail@web81402.mail.yahoo.com> References: <20050120205814.70166.qmail@web81402.mail.yahoo.com> Message-ID: <41F02178.1080606@pathology.washington.edu> We had an incident where a patient report went to a Barnes and Noble bookstore. At the time of the incident many users had access to our database for inputting new referring physicians. It is very easy to make a typo and the fax is going who knows where. The bookstore contacted us regarding the incident. We confirmed that we had their fax number in our database. An incident report was filed with risk management. Since the incident, we have restricted access to our database. Our facility recommendation is to verify fax numbers quarterly. With our staffing levels this is not feasible. We do send out annual verifications, that whoever signs the form is stating the fax number is correct and the machine is in a secure location. We will make several attempts to get the verification back. If there is no response then we will stop faxing to them and send a print copy through the mail. We have custom reports created to query the database when a physician is listed on a current case and their fax verification is more than a year old. Our facility policy states that a reasonable effort will be made to verify a fax number. What happens when a doctors office calls and asks for a copy of a patient report? We document in the database who we sent a copy of the report to. Victor Georgia Stewart wrote: >Hello and Good Afternoon Everyone > >In this era of HIPPA rules and regulations in the United States, I have not seen any comment on HIPPA as it relates to fax transmittal's of reports that might go to the wrong person/facility. If a report goes to the the wrong person/facility, isn't this is a breach of patient confidentiality covered by HIPPA? How are you handling faxs that go, or might go, to the wrong person/facility? > >Appreciate your comments. > >Georgia Stewart, BS, HTL > > > >Georgia >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. From mprice26 <@t> juno.com Thu Jan 20 16:04:33 2005 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] RE: Rudeness on Histonet Message-ID: <20050120.140511.23783.21286@webmail26.nyc.untd.com> I have been treated rudely also. I have e-mailed the histonet for a vendor's # or for other info and I will have people rerspond back with try google search. I know how to search for something via google or yellowpages.com but prefer to use the histonet because of the vast number of vendors that subscribe to the histonet. So not only do I receive the # I am asking for I also receive info from other vendors that I would not receive if I look it up on google. I wish those that think I am an idiot for not using google would just not respond to my request. If it bothers them so much that I am requesting info via the Histonet. It takes much less time to hit their delete button than it does for them to e-mail me a rude, condescending message. I would like to add that for the most part I receive prompt useful replys, it is just a few people that reply with rude answers. Marsha Price From paw555 <@t> yahoo.com Thu Jan 20 16:13:17 2005 From: paw555 <@t> yahoo.com (pam plumlee) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] GLP lab staining question Message-ID: <20050120221317.48866.qmail@web11605.mail.yahoo.com> Hello fellow histonetters: I'm setting up a special stain control/slide bank for our research pharm lab. I'm curious how special stain controls are validated. Is it as simple as screening the stained slides/blocks as they are cut and determining if they are positive or not, along with the proper documentation? Anyone care to share info? Thanks, PP __________________________________ Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. http://mobile.yahoo.com/maildemo From gcallis <@t> montana.edu Thu Jan 20 16:14:25 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] certification requirements to work in histo labs In-Reply-To: <20050120204937.7153.qmail@web81008.mail.yahoo.com> References: <20050120204937.7153.qmail@web81008.mail.yahoo.com> Message-ID: <6.0.0.22.1.20050120150521.01b1fbc0@gemini.msu.montana.edu> I am curious or rather need clarification here. Licensed means licensed by the state but HT(SCP) is a certification granted by a certification agency, ASCP/CAP. I don't think ASCP/CAP is governed by any state's' licensing regulations. Many moons ago, in New York, one did not have to be certified as an HT to be licensed by the state (one had to take a test for NY licensure), and there were NY licensed technicians without HT(ASCP) certification performing tech duties and, in fact, was the supervisor of our histopathology lab. At 01:49 PM 1/20/2005, you wrote: >Fran, > >In Florida where I am, to perform tech duties you must be licensed. >However, you may employ non techs to work as lab aides. These aides may >perform a wide variety of duties that usually are done by techs. >Specifically what they cannot do is: embed, cut, stain, frozen sections >etc. What they can do is: change reagents, coverslip, start and stop an >automated stainer. I hope this helps. > >Dawn Cowie, HT > >Pensacola Path > >_______________________________________________ >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 llewllew <@t> shaw.ca Thu Jan 20 16:36:24 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] GLP lab staining question References: <20050120221317.48866.qmail@web11605.mail.yahoo.com> Message-ID: <001601c4ff40$7923b6b0$80004246@yourlk4rlmsu> The way I was taught was to cut a series of sections, 50 or so, then stain the first and the last to make sure they were positive If they both are, it is presumed that all the others are also. If not, stain intermediate sections until one is positive, then use from the first one to that. Bryan Llewellyn ----- Original Message ----- From: "pam plumlee" To: "HistoNet Server" Sent: Thursday, January 20, 2005 2:13 PM Subject: [Histonet] GLP lab staining question > Hello fellow histonetters: I'm setting up a special > stain control/slide bank for our research pharm lab. > I'm curious how special stain controls are validated. > Is it as simple as screening the stained slides/blocks > as they are cut and determining if they are positive > or not, along with the proper documentation? Anyone > care to share info? Thanks, PP > > > > __________________________________ > Do you Yahoo!? > Take Yahoo! Mail with you! Get it on your mobile phone. > http://mobile.yahoo.com/maildemo > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ljones <@t> pathology.umsmed.edu Thu Jan 20 16:45:10 2005 From: ljones <@t> pathology.umsmed.edu (Linda Jones) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Special stain Message-ID: How do you charge for certain stains? Should charge due to the length of time it take to do the stain or the price of the chemicals? thanks Linda Harper-Jones BS.,HT/ HTL(ASCP) University of Mississippi Medical Center Department of Pathology Chief Histotechnologist Supervisor (601) 984-1576 (601) 984-4968 Fax ljones@pathology.umsmed.edu From JMacDonald <@t> mtsac.edu Thu Jan 20 17:49:45 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Special stain In-Reply-To: Message-ID: Special stains are based on the type of stain. For billing purposes there are two types: microorganism or not. 88312 and 88313. Cost or length of time is not a factor when billing. "Linda Jones" Sent by: histonet-bounces@lists.utsouthwestern.edu 01/20/2005 02:45 PM To cc Subject [Histonet] Special stain How do you charge for certain stains? Should charge due to the length of time it take to do the stain or the price of the chemicals? thanks Linda Harper-Jones BS.,HT/ HTL(ASCP) University of Mississippi Medical Center Department of Pathology Chief Histotechnologist Supervisor (601) 984-1576 (601) 984-4968 Fax ljones@pathology.umsmed.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kim.Osullivan <@t> med.monash.edu.au Thu Jan 20 22:04:26 2005 From: Kim.Osullivan <@t> med.monash.edu.au (Kim O'Sullivan) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Bouins fixation problem Message-ID: <130.194.114.210.1106279796.85555@my.monash.edu.au> Hi everybody, We have used bouin's fixative for fixing mouse kidneys for years with no problem, and have recently been experiencing problems cutting them once they have been processed and wax embedded. The kidney tissue has holes in it in the medulla region, is very difficult to cut(tissue brittle) and it appears almost like the wax has not infiltrated the kidneys properly. Additonally once the sections have been mounted on glass slides they appear fine but when we go to PAS stain them half of them come off the slides in solution (this has never happened before in the last 3 years)I have checked the processing schedule and that is fine (as is all the solutions) I have checked the temperature of the wax (which is fine). Does anyone out there know what our problem may be?? Additionally is there any way that any of the solutions used to make bouin's (picric acid, formalin and glacial acetic acid)can go off? And how would you check for this. Would appreciate any help!! Kim O'Sullivan From kmerriam2003 <@t> yahoo.com Fri Jan 21 07:24:15 2005 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] c-fos antibody in mouse tissue Message-ID: <20050121132416.20426.qmail@web52502.mail.yahoo.com> Hello All, I was wondering if anyone knew what tissue would make the best positive control for this antibody in mouse tissue. Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From akbitting <@t> geisinger.edu Fri Jan 21 08:23:30 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Coverslipping tape Message-ID: What is the concensus of opinion on Mercedes Medicals coverslipping tape? I got a sample to try and the girls I work with said they had it before, (I don't know how long ago) and in a couple days it started lifting. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 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 DPALLP <@t> aol.com Fri Jan 21 08:25:41 2005 From: DPALLP <@t> aol.com (DPALLP@aol.com) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] saturated fingertips Message-ID: <191.372a8459.2f226ae5@aol.com> Does anyone have any suggestions to alleviate water saturated fingertips from handling blocks from ice trays? Wearing gloves is too cumbersome and the different barrier lotions that we have tried do not seem to have an effect. Susie From mcauliff <@t> umdnj.edu Fri Jan 21 11:39:29 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Bouins fixation problem In-Reply-To: <130.194.114.210.1106279796.85555@my.monash.edu.au> References: <130.194.114.210.1106279796.85555@my.monash.edu.au> Message-ID: <41F13E51.1010507@umdnj.edu> Hi Kim: Hmm. Certainly sounds like an infiltration problem. My first though is that you are not getting all of the alcohol out of the medulla so xylene (or substitute) is not getting in so the wax is not getting in. When you say that you have checked all of the processing solutions and they are fine, how did you check them? What is the criteria for deciding that a solution is good? I would dump ALL of the processing solutions and ALL of the wax and start fresh. Run some kidneys, cut them, and see if that solves the problem. There is a small chance that the manufacturer of your wax has changed the formula but with modern waxes and additives such a change seems unlikely to cause problems. As for reagents going bad, picric acid last forever, formaldehyde is fine as long as there is no ppt. on the bottom of the bottle, I don't think glacial acetic acid goes bad. Even if the fixation was incomplete (you did not say how long you fix the kidneys, if you perfuse fixative, or if you slice them open) the alcohols should finish the fixation. Good luck! Geoff Kim O'Sullivan wrote: >Hi everybody, > >We have used bouin's fixative for fixing mouse kidneys for years with no problem, and have recently been experiencing problems cutting them once they have been processed and wax embedded. The kidney tissue has holes in it in the medulla region, is very difficult to cut(tissue brittle) and it appears almost like the wax has not infiltrated the kidneys properly. Additonally once the sections have been mounted on glass slides they appear fine but when we go to PAS stain them half of them come off the slides in solution (this has never happened before in the last 3 years)I have checked the processing schedule and that is fine (as is all the solutions) I have checked the temperature of the wax (which is fine). Does anyone out there know what our problem may be?? Additionally is there any way that any of the solutions used to make bouin's (picric acid, formalin and glacial acetic acid)can go off? And how would you check for this. > >Would appreciate any help!! > >Kim O'Sullivan > >_______________________________________________ >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 Michael.Rice <@t> holy-cross.com Fri Jan 21 09:00:37 2005 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Coverslipping tape Message-ID: <3BC92F29BE821745AB15E04C98EE028D693742@HCH2KMAIL.holy-cross.com> I have been using it for several months now and have seen no evidence of lifting, not to say that it wont happen in the future. A friend of mine has beeen using it for several years now with no problems Mike Rice Holy Cross hospital ft lauderdale -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Friday, January 21, 2005 9:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Coverslipping tape What is the concensus of opinion on Mercedes Medicals coverslipping tape? I got a sample to try and the girls I work with said they had it before, (I don't know how long ago) and in a couple days it started lifting. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- From GDawson <@t> dynacaremilwaukee.com Fri Jan 21 09:01:05 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] RE: Rudeness on Histonet Message-ID: Rude people on a listserver like this are unavoidable. Unfortunately, there is no "pre-membership rudeness testing". If you are tired of being chewed on like a bloody wildabeast in the midst of a den of lions, do what I was forced to do and avoid lengthy general postings as much as possible. Submit a pithy comment/question and pursue only those who seem to genuinely want to help and delete those who are jerks. Trying to rail against rudeness can become a full time job which doesn't pay anything. Histonet is too valuable a tool to just get rid of, though, at times, it is tempting. Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI From KarBieber <@t> aol.com Fri Jan 21 09:10:31 2005 From: KarBieber <@t> aol.com (KarBieber@aol.com) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Use of Sta-On for Immuno stains Message-ID: <194.36bbc139.2f227567@aol.com> I was just wondering how other labs are handling this situation: For prostate and breast core biopsies we cut extra slides for potential immuno staining, between the levels. The problem is the use of adhesive in the water bath. Since we are cutting H&E's, there is the possibility of fall off during staining if we DON'T use sta-on. But using it increases the possibility of fall off during immunos staining. And if we don't use the sta-on, we run the risk of fall off on ALL the cases the techs are cutting that day. I know there are lots of labs out there performing immunos, so I'd like to know how you've resolved this. Thanks, Karen From Terry.Marshall <@t> rothgen.nhs.uk Fri Jan 21 09:12:23 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Coverslipping tape Message-ID: The impression I get from my very limited experience, but mostly from this forum, is that it will happen, the only question being when. 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: Rice, Michael [mailto:Michael.Rice@holy-cross.com] Sent: 21 January 2005 15:01 To: Angela Bitting; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Coverslipping tape I have been using it for several months now and have seen no evidence of lifting, not to say that it wont happen in the future. A friend of mine has beeen using it for several years now with no problems Mike Rice Holy Cross hospital ft lauderdale -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Friday, January 21, 2005 9:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Coverslipping tape What is the concensus of opinion on Mercedes Medicals coverslipping tape? I got a sample to try and the girls I work with said they had it before, (I don't know how long ago) and in a couple days it started lifting. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From contact <@t> excaliburpathology.com Fri Jan 21 09:34:09 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Saturated fingertips Message-ID: <20050121153410.64391.qmail@web50305.mail.yahoo.com> What about picking the block up with forceps and blotting on a paper towel before handling? Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From HornHV <@t> archildrens.org Fri Jan 21 09:35:50 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Coverslipping tape Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6322BA44@EMAIL.archildrens.org> We do not use coverslipping tape, but we do get slides coverslipped with tape for consults. And all of the ones I have seen have lifted a bit if not all the way depending on how old the slides are. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr, Consultant Histopathologist Sent: Friday, January 21, 2005 9:12 AM To: Rice, Michael; Angela Bitting; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Coverslipping tape The impression I get from my very limited experience, but mostly from this forum, is that it will happen, the only question being when. 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: Rice, Michael [mailto:Michael.Rice@holy-cross.com] Sent: 21 January 2005 15:01 To: Angela Bitting; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Coverslipping tape I have been using it for several months now and have seen no evidence of lifting, not to say that it wont happen in the future. A friend of mine has beeen using it for several years now with no problems Mike Rice Holy Cross hospital ft lauderdale -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Friday, January 21, 2005 9:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Coverslipping tape What is the concensus of opinion on Mercedes Medicals coverslipping tape? I got a sample to try and the girls I work with said they had it before, (I don't know how long ago) and in a couple days it started lifting. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From Barry.R.Rittman <@t> uth.tmc.edu Fri Jan 21 09:35:59 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Rudeness Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0012C936B@UTHEVS3.mail.uthouston.edu> Not wishing to prolong the debate but... As I get older I realize that I had some misconceptions earlier in my life (I have different ones now). I used to use terms such as common courtesy and common sense. Now I am not so sure these are true terms. I believe that it is common courtesy to hold a door open for someone no matter what gender, age, ethnicity (or dress sense). It is also common courtesy to thank the person holding the door open. It is common courtesy not to block the grocery aisle when talking to another person, etc., etc. These are however things that are taught and while easy to learn are often not taught. Unfortunately in many cases individuals do not think about what they are doing and these "niceties" are often ignored. Common sense is also in many cases a misconception. This is evident by the Darwinian awards and other examples. I remember seeing an individual some years ago weighing out 20 gms of EDTA from a new 500 gm. bottle. Half the bottle was on the balance pan. No dah! I refrained from using the initial words that cast doubt on the genetic makeup of the individual and pointed out their error in a kind and diplomatic manner that first time. Mr. John Linder the senior technician who trained my wife and I, taught us to think carefully about everything that we did in the lab. This he regarded as the most important part of our training. In our modern world we spend too much time carrying out tasks and too little time thinking about what we are doing. I believe that thinking carefully about what we are doing to a large extent is responsible for what we perceive as "common sense". The world is becoming so computer dependent that often we do not look carefully at what we write or what we do. (I did spell check this article but also read it before sending out). Think about what would happen if the cash registers in stores were suddenly to be eliminated - all trade would immediately cease. As you can tell I've been thinking, but cannot guarantee it will result in my having more common sense. Have a great weekend (only if you want and are able to!!). Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rice, Michael Sent: Friday, January 21, 2005 9:01 AM To: Angela Bitting; histonet@lists.utsouthwestern.edu Subject: [SPAM] - RE: [Histonet] Coverslipping tape - Found word(s) list error in the Text body I have been using it for several months now and have seen no evidence of lifting, not to say that it wont happen in the future. A friend of mine has beeen using it for several years now with no problems Mike Rice Holy Cross hospital ft lauderdale -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Friday, January 21, 2005 9:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Coverslipping tape What is the concensus of opinion on Mercedes Medicals coverslipping tape? I got a sample to try and the girls I work with said they had it before, (I don't know how long ago) and in a couple days it started lifting. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TMcNemar <@t> lmhealth.org Fri Jan 21 09:37:30 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Use of Sta-On for Immuno stains Message-ID: <6CD94D97ED7D924BA5C2B588FA952821396778@nt_exchange.lmhealth.org> How did you determine that it was the Sta-on that caused your section to fall off? We have been using Sta-on for all slides for many, many years and have been doing IHC for 10 or so years without a problem. If you are doing HIER, you will need positively charged slide in addition to the Sta-on. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: KarBieber@aol.com [mailto:KarBieber@aol.com] Sent: Friday, January 21, 2005 10:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Use of Sta-On for Immuno stains I was just wondering how other labs are handling this situation: For prostate and breast core biopsies we cut extra slides for potential immuno staining, between the levels. The problem is the use of adhesive in the water bath. Since we are cutting H&E's, there is the possibility of fall off during staining if we DON'T use sta-on. But using it increases the possibility of fall off during immunos staining. And if we don't use the sta-on, we run the risk of fall off on ALL the cases the techs are cutting that day. I know there are lots of labs out there performing immunos, so I'd like to know how you've resolved this. Thanks, Karen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From plucas <@t> biopath.org Fri Jan 21 09:37:49 2005 From: plucas <@t> biopath.org (Paula Lucas) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Looking for Crystals Message-ID: <004301c4ffcf$2a24bb60$7c01a8c0@biopath.org> Hello all, When a specimen is sent for crystals, as in a case of joint fluid or gout, should the specimen be sent to us fresh or should it be in some sort of fixative. I had a discussion with the pathologist, and I told him that the specimen should be received fresh. He thought that fixing it in alcohol should be OK too. Paula Bio-Path Medical Group From funderwood <@t> mcohio.org Fri Jan 21 09:49:04 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:31 2005 Subject: [BULK] - [Histonet] saturated fingertips Message-ID: Finger cots might work well. Fred >>> 01/21/05 09:25AM >>> Does anyone have any suggestions to alleviate water saturated fingertips from handling blocks from ice trays? Wearing gloves is too cumbersome and the different barrier lotions that we have tried do not seem to have an effect. Susie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GDawson <@t> dynacaremilwaukee.com Fri Jan 21 09:49:55 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Use of Sta-On for Immuno stains Message-ID: Karen, I have found that sta-on doesn't produce alot of non-specific staining with IHC. It does, however, react with high molecular weight cytokeratins so what we did was to switch over to using charged slides for all cases so that we could forego the use of sta-on on stuff that the IHC techs cut themselves. If you look around, you should be able to find a reasonable price on charged slides. We still do IHC like AE1/AE3 & CK903's on the slides you mentioned which were precut by routine histology but I let the pathologists know what the situation is and they are able to interpret around the non-specific staining resulting from the sta-on. Good Luck, Glen Dawson -----Original Message----- From: KarBieber@aol.com [mailto:KarBieber@aol.com] Sent: Friday, January 21, 2005 9:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Use of Sta-On for Immuno stains I was just wondering how other labs are handling this situation: For prostate and breast core biopsies we cut extra slides for potential immuno staining, between the levels. The problem is the use of adhesive in the water bath. Since we are cutting H&E's, there is the possibility of fall off during staining if we DON'T use sta-on. But using it increases the possibility of fall off during immunos staining. And if we don't use the sta-on, we run the risk of fall off on ALL the cases the techs are cutting that day. I know there are lots of labs out there performing immunos, so I'd like to know how you've resolved this. Thanks, Karen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Fri Jan 21 09:53:57 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:24:31 2005 Subject: [BULK] - [Histonet] Looking for Crystals Message-ID: It might be safer to receive it fresh, then fix in absolute alcohol. That way you know it hasn't been placed in an aqueous solution. When processing, be sure to start in an absolute alcohol. Also avoid water when staining. Fred >>> "Paula Lucas" 01/21/05 10:37AM >>> Hello all, When a specimen is sent for crystals, as in a case of joint fluid or gout, should the specimen be sent to us fresh or should it be in some sort of fixative. I had a discussion with the pathologist, and I told him that the specimen should be received fresh. He thought that fixing it in alcohol should be OK too. Paula Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Fri Jan 21 09:59:08 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Use of Sta-On for Immuno stains Message-ID: We got rid of the Sta-on and started using charged slides for everything. The increase in volume, decreased the price of the charged slides. Sta-on will null the charge of the slides and hence the sections will fall off during HIER. In a perfect world this would be enough, but since we're not perfect, always check your lots of slides because sometimes the charge is just not there. The easiest way for us to tell, is by making sure the sections do not travel down the slide when you pick them up from the waterbath. Good luck! Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of KarBieber@aol.com Sent: Friday, January 21, 2005 9:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Use of Sta-On for Immuno stains I was just wondering how other labs are handling this situation: For prostate and breast core biopsies we cut extra slides for potential immuno staining, between the levels. The problem is the use of adhesive in the water bath. Since we are cutting H&E's, there is the possibility of fall off during staining if we DON'T use sta-on. But using it increases the possibility of fall off during immunos staining. And if we don't use the sta-on, we run the risk of fall off on ALL the cases the techs are cutting that day. I know there are lots of labs out there performing immunos, so I'd like to know how you've resolved this. Thanks, Karen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Jan 21 10:01:43 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Use of Sta-On for Immuno stains In-Reply-To: <194.36bbc139.2f227567@aol.com> References: <194.36bbc139.2f227567@aol.com> Message-ID: <6.0.0.22.1.20050121085607.01b90c48@gemini.msu.montana.edu> We bite the bullet, and buy Plus charge slides for ALL tissues and use distilled water in flotation bath. OR Polysine (Erie Scientific) - the poly l lysine commercial editions which allow one to manipulate sections a tidge onto slide surface. Maybe a tidge more expense but with shopping around for a good discount, vendors are usually very good about this, it eliminates a lot of frustration and recuts. Never looked back and everyone is happy. At 08:10 AM 1/21/2005, you wrote: >I was just wondering how other labs are handling this situation: For >prostate and breast core biopsies we cut extra slides for potential immuno >staining, >between the levels. The problem is the use of adhesive in the water bath. >Since we are cutting H&E's, there is the possibility of fall off during >staining >if we DON'T use sta-on. But using it increases the possibility of fall off >during immunos staining. And if we don't use the sta-on, we run the risk of >fall off on ALL the cases the techs are cutting that day. > >I know there are lots of labs out there performing immunos, so I'd like to >know how you've resolved this. >Thanks, >Karen 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 ploykasek <@t> phenopath.com Fri Jan 21 10:13:22 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Use of Sta-On for Immuno stains In-Reply-To: <194.36bbc139.2f227567@aol.com> Message-ID: I had a similar situation in a previous job. All H&Es, plus unstained levels were cut by the main histology lab, and they used sta-on in the water bath. I never had a problem with section fall off during immunos or non-specific staining. Perhaps this is dependent on the amount of sta-on used in the water bath? The majority of immunos I did cut myself, using a water bath with only water in it. Just my experience. Patti Loykasek PhenoPath Laboratories> I was just wondering how other labs are handling this situation: For > prostate and breast core biopsies we cut extra slides for potential immuno > staining, > between the levels. The problem is the use of adhesive in the water bath. > Since we are cutting H&E's, there is the possibility of fall off during > staining > if we DON'T use sta-on. But using it increases the possibility of fall off > during immunos staining. And if we don't use the sta-on, we run the risk of > fall off on ALL the cases the techs are cutting that day. > > I know there are lots of labs out there performing immunos, so I'd like to > know how you've resolved this. > Thanks, > Karen > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Fri Jan 21 10:18:07 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] IHC tech position open Message-ID: I would like to post the following job opening that we have at PhenoPath Laboratories. If you would like any further information, please contact me or Paul Moore. Thanks. Patti Loykasek PhenoPath Laboratories Seattle, WA IMMUNOHISTOCHEMISTRY TECHNOLOGIST PhenoPath Laboratories, a pathology reference laboratory, has an opportunity in the clinical immunohistochemistry division for a full-time technologist, day shift. We are in a state-of-the-art facility located on the scenic Ship Canal in the eclectic Fremont neighborhood of Seattle, WA. Job Description: Responsibilities may include performing immunohistochemistry, immunofluorescence, molecular testing on patient samples and tissue sectioning (paraffin and frozen). Participation in the development of new tests or technologies, as well as participation in clinical research projects are also included. Quality control, safety, and preventive maintenance procedures and documentation are required elements of this position as well. Required Skills/Experience: Strong preference given to ASCP certified (or certification-eligible) laboratory techs. Trained laboratory techs of any discipline are encouraged to apply (histotech, med tech, cytotech?..). PhenoPath Laboratories is committed to hiring the best person for the job. PhenoPath Laboratories is a national specialty pathology laboratory with several pathologists and technologists committed to providing outstanding clinical and research services. We offer an excellent compensation and benefits package, including 401k. To apply, please e-mail or fax completed application for employment and resume to: Paul Moore, Assistant to the Director of Human Resources PhenoPath Laboratories 551 N. 34th St., Suite 100 Seattle, WA 98103 Phone: 206 374-9000 Fax: 206 374-9009 E-mail: pmoore@phenopath.com From AMiller <@t> Elliot-HS.org Fri Jan 21 10:27:09 2005 From: AMiller <@t> Elliot-HS.org (Miller, Ann-Marie) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] RE: Histonet Digest, Vol 14, Issue 26 Message-ID: <6EE8A0B96E21D811BF440002557CEFDDCD96AB@exchange.elliot-hs.org> Karen, We save intervening levels between our prostates and breasts as well. One tech will cut those first with just distilled water on positively charged slides. Then he'll add gelatin to the water and cut everything else. Hope this helps. Ann -----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 14, Issue 26 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. NIH image J surface area unit (Subratab) 2. Certification requirements (Fran Lemons) 3. frozens on fat to image GFP (Patsy Ruegg) 4. Rudeness on histonet (Randolph-Habecker, Julie) 5. Re: NIH image J surface area unit (Rocan) 6. Re: Certification requirements (Gareth Davis) 7. Re: Rudeness on histonet (Gareth Davis) 8. certification requirements to work in histo labs (Dawn Cowie) 9. Re: Rudeness on histonet (Robyn Vazquez) 10. HIPPA rules and fax transmittals going to wrong person (Georgia Stewart) 11. RE: perfusion suggestions (Rittman, Barry) 12. Re: HIPPA rules and fax transmittals going to wrong person (Victor Tobias) 13. RE: Rudeness on Histonet (mprice26@juno.com) 14. GLP lab staining question (pam plumlee) 15. Re: certification requirements to work in histo labs (Gayle Callis) 16. Re: GLP lab staining question (Bryan Llewellyn) 17. Special stain (Linda Jones) 18. Re: Special stain (Jennifer MacDonald) 19. Bouins fixation problem (Kim O'Sullivan) 20. c-fos antibody in mouse tissue (Kim Merriam) 21. Coverslipping tape (Angela Bitting) 22. saturated fingertips (DPALLP@aol.com) 23. Re: Bouins fixation problem (Geoff McAuliffe) 24. RE: Coverslipping tape (Rice, Michael) 25. RE: Rudeness on Histonet (Dawson, Glen) 26. Use of Sta-On for Immuno stains (KarBieber@aol.com) ---------------------------------------------------------------------- Message: 1 Date: Fri, 21 Jan 2005 0:02:21 +0600 From: Subratab Subject: [Histonet] NIH image J surface area unit To: Message-ID: <200501201804.j0KI4YYb008313@mailout.proshikanet.com> Content-Type: text/plain; charset="iso-8859-1"; Dear Histo experts My friend is measuring glomerular surface area by using Image J software. He is in trouble to get the unit of the area calculated by the software, I mean, if the calculated area is to be expressed in square milimeter or square micrometer. He is capturing the figures from microscope with X100 magnification (oil-emersion lense) and then analyzing the glomerular surface area by the software in a computer. Could anybody please help to solve this. Sincerely Subrata Biswas University of Campinas SP, Brazil. -- http://www.histosearch.com/homepages/DrSubrataKumarBiswas.html ------------------------------ Message: 2 Date: Thu, 20 Jan 2005 13:26:57 -0500 From: "Fran Lemons" Subject: [Histonet] Certification requirements To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Has anyone out there seen anything in writing about not being able to work in histology labs without HT certification? Thanks Fran Walker ------------------------------ Message: 3 Date: Thu, 20 Jan 2005 12:16:05 -0700 From: "Patsy Ruegg" Subject: [Histonet] frozens on fat to image GFP To: Message-ID: <200501201916.j0KJG02p023804@chip.viawest.net> Content-Type: text/plain; charset="US-ASCII" Help! Here is the situation. An investigator wants me to cut fat tissue (briefly paraformaldehyde fixed,snap frozen in OCT) which does not cut well, so I had the brillant (I thought) idea of using the Instrumedics tape transfer system. I cut the sections and transfered them to the coated slides, air dryed them, and they looked pretty good. Since they want to preserve the fat I thought I would just use some aqueous mount and put a cs on them so they could look at the sections under UV. Well there was some kind of reaction from the ?water or glycerine or something that turned white so we can't see the tissue. Tryed soaking the slides in water to remove mount, the cs came off put the white reaction did not. so is the water reacting with the polymer coating (?might be GMA). Should I take these sections thru solvent anyway to get them coverslipped so they are clear??? Patsy ------------------------------ Message: 4 Date: Thu, 20 Jan 2005 11:27:09 -0800 From: "Randolph-Habecker, Julie" Subject: [Histonet] Rudeness on histonet To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAE7E@wala01.seattlecca.org> Content-Type: text/plain Dear Kristen and others: I too feel that Kristen was treated rudely. I have noticed this problem from time to time on HistoNet and I feel I have to say something. Some people generally assume that because you ask an open question that you are clueless. While you might get better answers by being more specific, the better way to respond to "any suggestions" is to ask if you could provide more details. Just assuming that you and your boss have no idea what you are doing and have ventured into this project naively is not fair. That response was not helpful - it was nothing but rude. Did they suggest a class or a book that they found practically good? No! The response was condescending and pointless. Please ask for clarification before you humiliate a person. And please do not tell someone they deserved to be treated rudely - that is even worse! People write into histonet with questions because there is a wealth of knowledge amongst the subscribers. This is valuable information and constitutes years and years of experience. Keep in mind, all of our experiences have been different. Just because a person might not have experience in one area doesn't mean that they are not very knowledgeable in another. In other words, if you are rude to a person they might not want to share valuable information with you when you ask. Or worse, they might leave the Histonet and deprive all of us of their knowledge! I too asked an open question one time about processing tissue with plastic beads in it and received an incredibly condescending response instructing me on how to design an experiment. I have a Ph.D. for god's sake. I have spent 15 years designing experiments. I also saw a young student publicly filleted for cheating when her instructor encouraged her to send her inquiry out to the Histonet. Kristen, please don't leave the histonet because of one (incredibly) rude response. Take it for what it is - useless! Please hold out for helpful people that actually have information to contribute. Hang in there! 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 Message: 2 Date: Thu, 20 Jan 2005 11:44:27 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] perfusion suggestions follow up To: Kristen Reynolds Cc: histonet@lists.utsouthwestern.edu Message-ID: <41F00A1B.6060101@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Dear Kristen: Your original posting (see below) gave no indication that you were familiar with perfusion for LM or EM. You did not ask for fixative composition, you just asked for "any suggestions". Given the broad scope of your question, the advice you got WAS helpful. The fixative combination for wanting to do both on the same tissue IS out there, but you have to be more specific in your question. The correct fixative combinatio will depend on: 1. Are you doing histochemistry? If so, what reaction? Pre-embedding or post-embedding reaction? 2. Are you doing immunohistochemistry? If so, what antigen? Pre-embedding or post-embedding reaction? 3. What plastic are you embedding in? 4. What routine stains do you need to do? 5. What part of the brain will you be looking at? The brain is a big place. 6. Do you need to do LM and EM on the same piece of tissue/same cells? Advice on HistoNet is free. Some of it is quite good. Checking published papers in refereed journals and looking at the results is always a good idea, espcially given the cost of experimental animals. Geoff Kristen Reynolds wrote: I need advice on perfusion solutions for rat/monkey brains. I need to use the tissue for both light microscopy and EM. Any suggestions? >Wow, I can't believe how rude of a response I got. I >thought this was for helpful comments. I know how to >perfuse for light microscopy and EM. I just thought >the fixative combination for wanting to do both on the >same tissue might be out there. I guess I won't be >asking histonet anything anymore. > >__________________________________________________ > 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. ------------------------------ Message: 5 Date: Thu, 20 Jan 2005 11:28:47 -0800 From: Rocan Subject: Re: [Histonet] NIH image J surface area unit To: "'histonet@pathology.swmed.edu'" , Subratab Message-ID: <816F081F-6B19-11D9-8D0A-000A9589219E@mac.com> Content-Type: text/plain; charset=US-ASCII; format=flowed I guess there are a couple of ways to do this. I use a stage micrometer (you can google "stage micrometer" and find it on the pictures). I take a picture of the ruler imprinted. Typically these rulers have a one millimeter ruler and 100 divisions of 10 micrometers each. So, with the photo using the same scope and objective you go to any of these programs like NIH ImageJ and open the document (same size as the photos you are measuring). There is a command where you place the mouse on one end drag and click on the other end of the markings. So, if you drag it over ten marking you then instruct the program that that distance is 100 micrometers. Then using these settings you do your measurements and the measurements would then be very precise and will be in micrometers. I do this with a different program but I know you can do it with ImageJ. I hope this helps. Rocio ----- Dr.Rocio Sierra-Honigmann Director Engineered Wound Repair Laboratory Cedars Sinai Research Institute Davis 1091 310-423-1882 Honigmannr@cshs.org On Jan 20, 2005, at 10:02 AM, Subratab wrote: > Dear Histo experts > My friend is measuring glomerular surface area by using Image J > software. He > is in trouble to get the unit of the area calculated by the software, I > mean, if the calculated area is to be expressed in square milimeter or > square micrometer. > He is capturing the figures from microscope with X100 magnification > (oil-emersion lense) and then analyzing the glomerular surface area by > the > software in a computer. > Could anybody please help to solve this. > Sincerely > > Subrata Biswas > University of Campinas > SP, Brazil. > > -- > http://www.histosearch.com/homepages/DrSubrataKumarBiswas.html > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Thu, 20 Jan 2005 12:09:42 -0800 (PST) From: Gareth Davis Subject: Re: [Histonet] Certification requirements To: Fran Lemons , Histonet Message-ID: <20050120200942.71181.qmail@web52708.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Fran, I believe it's based on what state you work in. I know that Florida requires an ASCP certification as well as a Florida license. Tennessee, where I work, does not require it. Gareth Davis Fran Lemons wrote: Has anyone out there seen anything in writing about not being able to work in histology labs without HT certification? Thanks Fran Walker _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' ------------------------------ Message: 7 Date: Thu, 20 Jan 2005 12:14:32 -0800 (PST) From: Gareth Davis Subject: Re: [Histonet] Rudeness on histonet To: "Randolph-Habecker, Julie" , Histonet Message-ID: <20050120201432.90980.qmail@web52710.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Here! Here! to Julie. I have also experienced the rudeness and seen others put down and treated as if they were stupid. It's very discouraging. Gareth "Randolph-Habecker, Julie" wrote: Dear Kristen and others: I too feel that Kristen was treated rudely. I have noticed this problem from time to time on HistoNet and I feel I have to say something. Some people generally assume that because you ask an open question that you are clueless. While you might get better answers by being more specific, the better way to respond to "any suggestions" is to ask if you could provide more details. Just assuming that you and your boss have no idea what you are doing and have ventured into this project naively is not fair. That response was not helpful - it was nothing but rude. Did they suggest a class or a book that they found practically good? No! The response was condescending and pointless. Please ask for clarification before you humiliate a person. And please do not tell someone they deserved to be treated rudely - that is even worse! People write into histonet with questions because there is a wealth of knowledge amongst the subscribers. This is valuable information and constitutes years and years of experience. Keep in mind, all of our experiences have been different. Just because a person might not have experience in one area doesn't mean that they are not very knowledgeable in another. In other words, if you are rude to a person they might not want to share valuable information with you when you ask. Or worse, they might leave the Histonet and deprive all of us of their knowledge! I too asked an open question one time about processing tissue with plastic beads in it and received an incredibly condescending response instructing me on how to design an experiment. I have a Ph.D. for god's sake. I have spent 15 years designing experiments. I also saw a young student publicly filleted for cheating when her instructor encouraged her to send her inquiry out to the Histonet. Kristen, please don't leave the histonet because of one (incredibly) rude response. Take it for what it is - useless! Please hold out for helpful people that actually have information to contribute. Hang in there! 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 Message: 2 Date: Thu, 20 Jan 2005 11:44:27 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] perfusion suggestions follow up To: Kristen Reynolds Cc: histonet@lists.utsouthwestern.edu Message-ID: <41F00A1B.6060101@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Dear Kristen: Your original posting (see below) gave no indication that you were familiar with perfusion for LM or EM. You did not ask for fixative composition, you just asked for "any suggestions". Given the broad scope of your question, the advice you got WAS helpful. The fixative combination for wanting to do both on the same tissue IS out there, but you have to be more specific in your question. The correct fixative combinatio will depend on: 1. Are you doing histochemistry? If so, what reaction? Pre-embedding or post-embedding reaction? 2. Are you doing immunohistochemistry? If so, what antigen? Pre-embedding or post-embedding reaction? 3. What plastic are you embedding in? 4. What routine stains do you need to do? 5. What part of the brain will you be looking at? The brain is a big place. 6. Do you need to do LM and EM on the same piece of tissue/same cells? Advice on HistoNet is free. Some of it is quite good. Checking published papers in refereed journals and looking at the results is always a good idea, espcially given the cost of experimental animals. Geoff Kristen Reynolds wrote: I need advice on perfusion solutions for rat/monkey brains. I need to use the tissue for both light microscopy and EM. Any suggestions? >Wow, I can't believe how rude of a response I got. I >thought this was for helpful comments. I know how to >perfuse for light microscopy and EM. I just thought >the fixative combination for wanting to do both on the >same tissue might be out there. I guess I won't be >asking histonet anything anymore. > >__________________________________________________ > 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 --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' ------------------------------ Message: 8 Date: Thu, 20 Jan 2005 12:49:37 -0800 (PST) From: Dawn Cowie Subject: [Histonet] certification requirements to work in histo labs To: histonet Message-ID: <20050120204937.7153.qmail@web81008.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Fran, In Florida where I am, to perform tech duties you must be licensed. However, you may employ non techs to work as lab aides. These aides may perform a wide variety of duties that usually are done by techs. Specifically what they cannot do is: embed, cut, stain, frozen sections etc. What they can do is: change reagents, coverslip, start and stop an automated stainer. I hope this helps. Dawn Cowie, HT Pensacola Path ------------------------------ Message: 9 Date: Thu, 20 Jan 2005 12:57:54 -0800 From: "Robyn Vazquez" Subject: Re: [Histonet] Rudeness on histonet To: histonet@lists.utsouthwestern.edu, jhabecke@seattlecca.org Message-ID: Content-Type: text/plain; charset=us-ascii BRAVO ladies for speaking your mind!!!! :0) Robyn >>> "Randolph-Habecker, Julie" 1/20/2005 11:27:09 AM >>> Dear Kristen and others: I too feel that Kristen was treated rudely. I have noticed this problem from time to time on HistoNet and I feel I have to say something. Some people generally assume that because you ask an open question that you are clueless. While you might get better answers by being more specific, the better way to respond to "any suggestions" is to ask if you could provide more details. Just assuming that you and your boss have no idea what you are doing and have ventured into this project naively is not fair. That response was not helpful - it was nothing but rude. Did they suggest a class or a book that they found practically good? No! The response was condescending and pointless. Please ask for clarification before you humiliate a person. And please do not tell someone they deserved to be treated rudely - that is even worse! People write into histonet with questions because there is a wealth of knowledge amongst the subscribers. This is valuable information and constitutes years and years of experience. Keep in mind, all of our experiences have been different. Just because a person might not have experience in one area doesn't mean that they are not very knowledgeable in another. In other words, if you are rude to a person they might not want to share valuable information with you when you ask. Or worse, they might leave the Histonet and deprive all of us of their knowledge! I too asked an open question one time about processing tissue with plastic beads in it and received an incredibly condescending response instructing me on how to design an experiment. I have a Ph.D. for god's sake. I have spent 15 years designing experiments. I also saw a young student publicly filleted for cheating when her instructor encouraged her to send her inquiry out to the Histonet. Kristen, please don't leave the histonet because of one (incredibly) rude response. Take it for what it is - useless! Please hold out for helpful people that actually have information to contribute. Hang in there! 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 Message: 2 Date: Thu, 20 Jan 2005 11:44:27 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] perfusion suggestions follow up To: Kristen Reynolds Cc: histonet@lists.utsouthwestern.edu Message-ID: <41F00A1B.6060101@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Dear Kristen: Your original posting (see below) gave no indication that you were familiar with perfusion for LM or EM. You did not ask for fixative composition, you just asked for "any suggestions". Given the broad scope of your question, the advice you got WAS helpful. The fixative combination for wanting to do both on the same tissue IS out there, but you have to be more specific in your question. The correct fixative combinatio will depend on: 1. Are you doing histochemistry? If so, what reaction? Pre-embedding or post-embedding reaction? 2. Are you doing immunohistochemistry? If so, what antigen? Pre-embedding or post-embedding reaction? 3. What plastic are you embedding in? 4. What routine stains do you need to do? 5. What part of the brain will you be looking at? The brain is a big place. 6. Do you need to do LM and EM on the same piece of tissue/same cells? Advice on HistoNet is free. Some of it is quite good. Checking published papers in refereed journals and looking at the results is always a good idea, espcially given the cost of experimental animals. Geoff Kristen Reynolds wrote: I need advice on perfusion solutions for rat/monkey brains. I need to use the tissue for both light microscopy and EM. Any suggestions? >Wow, I can't believe how rude of a response I got. I >thought this was for helpful comments. I know how to >perfuse for light microscopy and EM. I just thought >the fixative combination for wanting to do both on the >same tissue might be out there. I guess I won't be >asking histonet anything anymore. > >__________________________________________________ > 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 ------------------------------ Message: 10 Date: Thu, 20 Jan 2005 12:58:14 -0800 (PST) From: Georgia Stewart Subject: [Histonet] HIPPA rules and fax transmittals going to wrong person To: histonet Message-ID: <20050120205814.70166.qmail@web81402.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hello and Good Afternoon Everyone In this era of HIPPA rules and regulations in the United States, I have not seen any comment on HIPPA as it relates to fax transmittal's of reports that might go to the wrong person/facility. If a report goes to the the wrong person/facility, isn't this is a breach of patient confidentiality covered by HIPPA? How are you handling faxs that go, or might go, to the wrong person/facility? Appreciate your comments. Georgia Stewart, BS, HTL Georgia ------------------------------ Message: 11 Date: Thu, 20 Jan 2005 15:00:39 -0600 From: "Rittman, Barry" Subject: RE: [Histonet] perfusion suggestions To: "histonet" Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0012C92C3@UTHEVS3.mail.uthouston.edu> Content-Type: text/plain; charset="us-ascii" Kristen I am not trying to justify any response that you received but I think that I understand why the response was phrased in that particular manner. Many of the questions that arrive at Histonet are very specific, and generally get a lot of responses. The broader questions are often difficult to answer (and in general I personally do not respond to these) and often receive few responses. It is usually not possible to determine the level of knowledge of the individual who is asking the question and there may be a wide variety of answers. Here on Histonet we are often between a rock and a hard place. I believe that it is a natural tendency nowadays to pose the question in the shortest way possible on the assumption that nobody wishes to spend their time to read a long discourse. This unfortunately often results in a communication that is open to a wide variety of interpretations. One interpretation is that the individual posing the question wants to be informed about all aspects of a particular technique from anesthetizing an animal to interpreting the final result, rather than carrying out the initial groundwork themselves. There is usually no way to judge if this is true from the original communications. This is the era of rapid fire information but unfortunately the recipients do not always have the same background or experiences to view that information in the same light. In many cases I feel that in response to complex questions, individuals with the appropriate expertise should provide their telephone number so that a dialogue can be set up. While Histonet is very useful, a one on one dialogue generally will provide the most relevant information. I hope that you will continue to ask questions on Histonet. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Reynolds Sent: Thursday, January 20, 2005 9:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] perfusion suggestions Wow, I can't believe how rude of a response I got. I thought this was for helpful comments. I know how to perfuse for light microscopy and EM. I just thought the fixative combination for wanting to do both on the same tissue might be out there. I guess I won't be asking histonet anything anymore. __________________________________________________ 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 ------------------------------ Message: 12 Date: Thu, 20 Jan 2005 13:24:08 -0800 From: Victor Tobias Subject: Re: [Histonet] HIPPA rules and fax transmittals going to wrong person To: Georgia Stewart Cc: histonet Message-ID: <41F02178.1080606@pathology.washington.edu> Content-Type: text/plain; charset=us-ascii; format=flowed We had an incident where a patient report went to a Barnes and Noble bookstore. At the time of the incident many users had access to our database for inputting new referring physicians. It is very easy to make a typo and the fax is going who knows where. The bookstore contacted us regarding the incident. We confirmed that we had their fax number in our database. An incident report was filed with risk management. Since the incident, we have restricted access to our database. Our facility recommendation is to verify fax numbers quarterly. With our staffing levels this is not feasible. We do send out annual verifications, that whoever signs the form is stating the fax number is correct and the machine is in a secure location. We will make several attempts to get the verification back. If there is no response then we will stop faxing to them and send a print copy through the mail. We have custom reports created to query the database when a physician is listed on a current case and their fax verification is more than a year old. Our facility policy states that a reasonable effort will be made to verify a fax number. What happens when a doctors office calls and asks for a copy of a patient report? We document in the database who we sent a copy of the report to. Victor Georgia Stewart wrote: >Hello and Good Afternoon Everyone > >In this era of HIPPA rules and regulations in the United States, I have not seen any comment on HIPPA as it relates to fax transmittal's of reports that might go to the wrong person/facility. If a report goes to the the wrong person/facility, isn't this is a breach of patient confidentiality covered by HIPPA? How are you handling faxs that go, or might go, to the wrong person/facility? > >Appreciate your comments. > >Georgia Stewart, BS, HTL > > > >Georgia >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. ------------------------------ Message: 13 Date: Thu, 20 Jan 2005 22:04:33 GMT From: "mprice26@juno.com" Subject: [Histonet] RE: Rudeness on Histonet To: histonet@lists.utsouthwestern.edu Message-ID: <20050120.140511.23783.21286@webmail26.nyc.untd.com> Content-Type: text/plain I have been treated rudely also. I have e-mailed the histonet for a vendor's # or for other info and I will have people rerspond back with try google search. I know how to search for something via google or yellowpages.com but prefer to use the histonet because of the vast number of vendors that subscribe to the histonet. So not only do I receive the # I am asking for I also receive info from other vendors that I would not receive if I look it up on google. I wish those that think I am an idiot for not using google would just not respond to my request. If it bothers them so much that I am requesting info via the Histonet. It takes much less time to hit their delete button than it does for them to e-mail me a rude, condescending message. I would like to add that for the most part I receive prompt useful replys, it is just a few people that reply with rude answers. Marsha Price ------------------------------ Message: 14 Date: Thu, 20 Jan 2005 14:13:17 -0800 (PST) From: pam plumlee Subject: [Histonet] GLP lab staining question To: HistoNet Server Message-ID: <20050120221317.48866.qmail@web11605.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hello fellow histonetters: I'm setting up a special stain control/slide bank for our research pharm lab. I'm curious how special stain controls are validated. Is it as simple as screening the stained slides/blocks as they are cut and determining if they are positive or not, along with the proper documentation? Anyone care to share info? Thanks, PP __________________________________ Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. http://mobile.yahoo.com/maildemo ------------------------------ Message: 15 Date: Thu, 20 Jan 2005 15:14:25 -0700 From: Gayle Callis Subject: Re: [Histonet] certification requirements to work in histo labs To: Dawn Cowie , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050120150521.01b1fbc0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I am curious or rather need clarification here. Licensed means licensed by the state but HT(SCP) is a certification granted by a certification agency, ASCP/CAP. I don't think ASCP/CAP is governed by any state's' licensing regulations. Many moons ago, in New York, one did not have to be certified as an HT to be licensed by the state (one had to take a test for NY licensure), and there were NY licensed technicians without HT(ASCP) certification performing tech duties and, in fact, was the supervisor of our histopathology lab. At 01:49 PM 1/20/2005, you wrote: >Fran, > >In Florida where I am, to perform tech duties you must be licensed. >However, you may employ non techs to work as lab aides. These aides may >perform a wide variety of duties that usually are done by techs. >Specifically what they cannot do is: embed, cut, stain, frozen sections >etc. What they can do is: change reagents, coverslip, start and stop an >automated stainer. I hope this helps. > >Dawn Cowie, HT > >Pensacola Path > >_______________________________________________ >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: 16 Date: Thu, 20 Jan 2005 14:36:24 -0800 From: Bryan Llewellyn Subject: Re: [Histonet] GLP lab staining question To: Histonet Message-ID: <001601c4ff40$7923b6b0$80004246@yourlk4rlmsu> Content-Type: text/plain; reply-type=original; charset=iso-8859-1; format=flowed The way I was taught was to cut a series of sections, 50 or so, then stain the first and the last to make sure they were positive If they both are, it is presumed that all the others are also. If not, stain intermediate sections until one is positive, then use from the first one to that. Bryan Llewellyn ----- Original Message ----- From: "pam plumlee" To: "HistoNet Server" Sent: Thursday, January 20, 2005 2:13 PM Subject: [Histonet] GLP lab staining question > Hello fellow histonetters: I'm setting up a special > stain control/slide bank for our research pharm lab. > I'm curious how special stain controls are validated. > Is it as simple as screening the stained slides/blocks > as they are cut and determining if they are positive > or not, along with the proper documentation? Anyone > care to share info? Thanks, PP > > > > __________________________________ > Do you Yahoo!? > Take Yahoo! Mail with you! Get it on your mobile phone. > http://mobile.yahoo.com/maildemo > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Thu, 20 Jan 2005 16:45:10 -0600 From: "Linda Jones" Subject: [Histonet] Special stain To: Message-ID: Content-Type: text/plain; charset=US-ASCII How do you charge for certain stains? Should charge due to the length of time it take to do the stain or the price of the chemicals? thanks Linda Harper-Jones BS.,HT/ HTL(ASCP) University of Mississippi Medical Center Department of Pathology Chief Histotechnologist Supervisor (601) 984-1576 (601) 984-4968 Fax ljones@pathology.umsmed.edu ------------------------------ Message: 18 Date: Thu, 20 Jan 2005 15:49:45 -0800 From: Jennifer MacDonald Subject: Re: [Histonet] Special stain To: "Linda Jones" Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Special stains are based on the type of stain. For billing purposes there are two types: microorganism or not. 88312 and 88313. Cost or length of time is not a factor when billing. "Linda Jones" Sent by: histonet-bounces@lists.utsouthwestern.edu 01/20/2005 02:45 PM To cc Subject [Histonet] Special stain How do you charge for certain stains? Should charge due to the length of time it take to do the stain or the price of the chemicals? thanks Linda Harper-Jones BS.,HT/ HTL(ASCP) University of Mississippi Medical Center Department of Pathology Chief Histotechnologist Supervisor (601) 984-1576 (601) 984-4968 Fax ljones@pathology.umsmed.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Fri, 21 Jan 2005 04:04:26 +0000 From: Kim O'Sullivan Subject: [Histonet] Bouins fixation problem To: histonet@lists.utsouthwestern.edu Message-ID: <130.194.114.210.1106279796.85555@my.monash.edu.au> Content-Type: text/plain Hi everybody, We have used bouin's fixative for fixing mouse kidneys for years with no problem, and have recently been experiencing problems cutting them once they have been processed and wax embedded. The kidney tissue has holes in it in the medulla region, is very difficult to cut(tissue brittle) and it appears almost like the wax has not infiltrated the kidneys properly. Additonally once the sections have been mounted on glass slides they appear fine but when we go to PAS stain them half of them come off the slides in solution (this has never happened before in the last 3 years)I have checked the processing schedule and that is fine (as is all the solutions) I have checked the temperature of the wax (which is fine). Does anyone out there know what our problem may be?? Additionally is there any way that any of the solutions used to make bouin's (picric acid, formalin and glacial acetic acid)can go off? And how would you check for this. Would appreciate any help!! Kim O'Sullivan ------------------------------ Message: 20 Date: Fri, 21 Jan 2005 05:24:15 -0800 (PST) From: Kim Merriam Subject: [Histonet] c-fos antibody in mouse tissue To: Histonet Message-ID: <20050121132416.20426.qmail@web52502.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hello All, I was wondering if anyone knew what tissue would make the best positive control for this antibody in mouse tissue. Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 21 Date: Fri, 21 Jan 2005 09:23:30 -0500 From: "Angela Bitting" Subject: [Histonet] Coverslipping tape To: Message-ID: Content-Type: text/plain; charset=US-ASCII What is the concensus of opinion on Mercedes Medicals coverslipping tape? I got a sample to try and the girls I work with said they had it before, (I don't know how long ago) and in a couple days it started lifting. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 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: 22 Date: Fri, 21 Jan 2005 09:25:41 EST From: DPALLP@aol.com Subject: [Histonet] saturated fingertips To: histonet@pathology.swmed.edu Message-ID: <191.372a8459.2f226ae5@aol.com> Content-Type: text/plain; charset="US-ASCII" Does anyone have any suggestions to alleviate water saturated fingertips from handling blocks from ice trays? Wearing gloves is too cumbersome and the different barrier lotions that we have tried do not seem to have an effect. Susie ------------------------------ Message: 23 Date: Fri, 21 Jan 2005 09:39:29 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] Bouins fixation problem To: Kim O'Sullivan Cc: histonet@lists.utsouthwestern.edu Message-ID: <41F13E51.1010507@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Hi Kim: Hmm. Certainly sounds like an infiltration problem. My first though is that you are not getting all of the alcohol out of the medulla so xylene (or substitute) is not getting in so the wax is not getting in. When you say that you have checked all of the processing solutions and they are fine, how did you check them? What is the criteria for deciding that a solution is good? I would dump ALL of the processing solutions and ALL of the wax and start fresh. Run some kidneys, cut them, and see if that solves the problem. There is a small chance that the manufacturer of your wax has changed the formula but with modern waxes and additives such a change seems unlikely to cause problems. As for reagents going bad, picric acid last forever, formaldehyde is fine as long as there is no ppt. on the bottom of the bottle, I don't think glacial acetic acid goes bad. Even if the fixation was incomplete (you did not say how long you fix the kidneys, if you perfuse fixative, or if you slice them open) the alcohols should finish the fixation. Good luck! Geoff Kim O'Sullivan wrote: >Hi everybody, > >We have used bouin's fixative for fixing mouse kidneys for years with no problem, and have recently been experiencing problems cutting them once they have been processed and wax embedded. The kidney tissue has holes in it in the medulla region, is very difficult to cut(tissue brittle) and it appears almost like the wax has not infiltrated the kidneys properly. Additonally once the sections have been mounted on glass slides they appear fine but when we go to PAS stain them half of them come off the slides in solution (this has never happened before in the last 3 years)I have checked the processing schedule and that is fine (as is all the solutions) I have checked the temperature of the wax (which is fine). Does anyone out there know what our problem may be?? Additionally is there any way that any of the solutions used to make bouin's (picric acid, formalin and glacial acetic acid)can go off? And how would you check for this. > >Would appreciate any help!! > >Kim O'Sullivan > >_______________________________________________ >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: 24 Date: Fri, 21 Jan 2005 10:00:37 -0500 From: "Rice, Michael" Subject: RE: [Histonet] Coverslipping tape To: "Angela Bitting" , Message-ID: <3BC92F29BE821745AB15E04C98EE028D693742@HCH2KMAIL.holy-cross.com> Content-Type: text/plain I have been using it for several months now and have seen no evidence of lifting, not to say that it wont happen in the future. A friend of mine has beeen using it for several years now with no problems Mike Rice Holy Cross hospital ft lauderdale -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Friday, January 21, 2005 9:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Coverslipping tape What is the concensus of opinion on Mercedes Medicals coverslipping tape? I got a sample to try and the girls I work with said they had it before, (I don't know how long ago) and in a couple days it started lifting. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- ------------------------------ Message: 25 Date: Fri, 21 Jan 2005 09:01:05 -0600 From: "Dawson, Glen" Subject: [Histonet] RE: Rudeness on Histonet To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Rude people on a listserver like this are unavoidable. Unfortunately, there is no "pre-membership rudeness testing". If you are tired of being chewed on like a bloody wildabeast in the midst of a den of lions, do what I was forced to do and avoid lengthy general postings as much as possible. Submit a pithy comment/question and pursue only those who seem to genuinely want to help and delete those who are jerks. Trying to rail against rudeness can become a full time job which doesn't pay anything. Histonet is too valuable a tool to just get rid of, though, at times, it is tempting. Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI ------------------------------ Message: 26 Date: Fri, 21 Jan 2005 10:10:31 EST From: KarBieber@aol.com Subject: [Histonet] Use of Sta-On for Immuno stains To: histonet@lists.utsouthwestern.edu Message-ID: <194.36bbc139.2f227567@aol.com> Content-Type: text/plain; charset="US-ASCII" I was just wondering how other labs are handling this situation: For prostate and breast core biopsies we cut extra slides for potential immuno staining, between the levels. The problem is the use of adhesive in the water bath. Since we are cutting H&E's, there is the possibility of fall off during staining if we DON'T use sta-on. But using it increases the possibility of fall off during immunos staining. And if we don't use the sta-on, we run the risk of fall off on ALL the cases the techs are cutting that day. I know there are lots of labs out there performing immunos, so I'd like to know how you've resolved this. Thanks, Karen ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 14, Issue 26 **************************************** CONFIDENTIAL COMMUNICATION - PLEASE READ PRIVACY NOTICE This communication is confidential and may be read only by its intended recipient(s). It may contain legally privileged and protected information. If you believe you have received this communication in error, please "Reply" to the Sender and so indicate or call (603) 663-2800. Then, please promptly "Delete" this communication from your computer. This communication, and any information contained herein, may only be forwarded, printed, disclosed, copied or disseminated by those specifically authorized to do so. UNAUTHORIZED DISCLOSURE MAY RESULT IN LEGAL LIABILITY FOR THOSE PERSONS RESPONSIBLE. From Dorothy.L.Webb <@t> HealthPartners.Com Fri Jan 21 10:26:11 2005 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] RE: Histonet Digest, Vol 14, Issue 26 Message-ID: We have been using the Mercedes coverslipping tape and find it adhering very well. I switched this hospital over to it over a year ago due to the cost savings and we have no trouble with lifting, even the older slides! There are times that we have had a "splicing" withing the middle of the tape, but, they replaced those and not for a long time has this happened. The tape is slightly thicker, so you have to make certain your coverslipper is set to push the tape down with the right amount of pressure. Also, to the person cutting IHC's with the adhesive in it. We bought a small waterbath to keep sterile water in and this is where the IHC's get laid out on as well as out AFB's and Ahoramine Rhodamine stains. It has worked well for us! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Friday, January 21, 2005 9:17 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 14, Issue 26 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. NIH image J surface area unit (Subratab) 2. Certification requirements (Fran Lemons) 3. frozens on fat to image GFP (Patsy Ruegg) 4. Rudeness on histonet (Randolph-Habecker, Julie) 5. Re: NIH image J surface area unit (Rocan) 6. Re: Certification requirements (Gareth Davis) 7. Re: Rudeness on histonet (Gareth Davis) 8. certification requirements to work in histo labs (Dawn Cowie) 9. Re: Rudeness on histonet (Robyn Vazquez) 10. HIPPA rules and fax transmittals going to wrong person (Georgia Stewart) 11. RE: perfusion suggestions (Rittman, Barry) 12. Re: HIPPA rules and fax transmittals going to wrong person (Victor Tobias) 13. RE: Rudeness on Histonet (mprice26@juno.com) 14. GLP lab staining question (pam plumlee) 15. Re: certification requirements to work in histo labs (Gayle Callis) 16. Re: GLP lab staining question (Bryan Llewellyn) 17. Special stain (Linda Jones) 18. Re: Special stain (Jennifer MacDonald) 19. Bouins fixation problem (Kim O'Sullivan) 20. c-fos antibody in mouse tissue (Kim Merriam) 21. Coverslipping tape (Angela Bitting) 22. saturated fingertips (DPALLP@aol.com) 23. Re: Bouins fixation problem (Geoff McAuliffe) 24. RE: Coverslipping tape (Rice, Michael) 25. RE: Rudeness on Histonet (Dawson, Glen) 26. Use of Sta-On for Immuno stains (KarBieber@aol.com) ---------------------------------------------------------------------- Message: 1 Date: Fri, 21 Jan 2005 0:02:21 +0600 From: Subratab Subject: [Histonet] NIH image J surface area unit To: Message-ID: <200501201804.j0KI4YYb008313@mailout.proshikanet.com> Content-Type: text/plain; charset="iso-8859-1"; Dear Histo experts My friend is measuring glomerular surface area by using Image J software. He is in trouble to get the unit of the area calculated by the software, I mean, if the calculated area is to be expressed in square milimeter or square micrometer. He is capturing the figures from microscope with X100 magnification (oil-emersion lense) and then analyzing the glomerular surface area by the software in a computer. Could anybody please help to solve this. Sincerely Subrata Biswas University of Campinas SP, Brazil. -- http://www.histosearch.com/homepages/DrSubrataKumarBiswas.html ------------------------------ Message: 2 Date: Thu, 20 Jan 2005 13:26:57 -0500 From: "Fran Lemons" Subject: [Histonet] Certification requirements To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Has anyone out there seen anything in writing about not being able to work in histology labs without HT certification? Thanks Fran Walker ------------------------------ Message: 3 Date: Thu, 20 Jan 2005 12:16:05 -0700 From: "Patsy Ruegg" Subject: [Histonet] frozens on fat to image GFP To: Message-ID: <200501201916.j0KJG02p023804@chip.viawest.net> Content-Type: text/plain; charset="US-ASCII" Help! Here is the situation. An investigator wants me to cut fat tissue (briefly paraformaldehyde fixed,snap frozen in OCT) which does not cut well, so I had the brillant (I thought) idea of using the Instrumedics tape transfer system. I cut the sections and transfered them to the coated slides, air dryed them, and they looked pretty good. Since they want to preserve the fat I thought I would just use some aqueous mount and put a cs on them so they could look at the sections under UV. Well there was some kind of reaction from the ?water or glycerine or something that turned white so we can't see the tissue. Tryed soaking the slides in water to remove mount, the cs came off put the white reaction did not. so is the water reacting with the polymer coating (?might be GMA). Should I take these sections thru solvent anyway to get them coverslipped so they are clear??? Patsy ------------------------------ Message: 4 Date: Thu, 20 Jan 2005 11:27:09 -0800 From: "Randolph-Habecker, Julie" Subject: [Histonet] Rudeness on histonet To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EAE7E@wala01.seattlecca.org> Content-Type: text/plain Dear Kristen and others: I too feel that Kristen was treated rudely. I have noticed this problem from time to time on HistoNet and I feel I have to say something. Some people generally assume that because you ask an open question that you are clueless. While you might get better answers by being more specific, the better way to respond to "any suggestions" is to ask if you could provide more details. Just assuming that you and your boss have no idea what you are doing and have ventured into this project naively is not fair. That response was not helpful - it was nothing but rude. Did they suggest a class or a book that they found practically good? No! The response was condescending and pointless. Please ask for clarification before you humiliate a person. And please do not tell someone they deserved to be treated rudely - that is even worse! People write into histonet with questions because there is a wealth of knowledge amongst the subscribers. This is valuable information and constitutes years and years of experience. Keep in mind, all of our experiences have been different. Just because a person might not have experience in one area doesn't mean that they are not very knowledgeable in another. In other words, if you are rude to a person they might not want to share valuable information with you when you ask. Or worse, they might leave the Histonet and deprive all of us of their knowledge! I too asked an open question one time about processing tissue with plastic beads in it and received an incredibly condescending response instructing me on how to design an experiment. I have a Ph.D. for god's sake. I have spent 15 years designing experiments. I also saw a young student publicly filleted for cheating when her instructor encouraged her to send her inquiry out to the Histonet. Kristen, please don't leave the histonet because of one (incredibly) rude response. Take it for what it is - useless! Please hold out for helpful people that actually have information to contribute. Hang in there! 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 Message: 2 Date: Thu, 20 Jan 2005 11:44:27 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] perfusion suggestions follow up To: Kristen Reynolds Cc: histonet@lists.utsouthwestern.edu Message-ID: <41F00A1B.6060101@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Dear Kristen: Your original posting (see below) gave no indication that you were familiar with perfusion for LM or EM. You did not ask for fixative composition, you just asked for "any suggestions". Given the broad scope of your question, the advice you got WAS helpful. The fixative combination for wanting to do both on the same tissue IS out there, but you have to be more specific in your question. The correct fixative combinatio will depend on: 1. Are you doing histochemistry? If so, what reaction? Pre-embedding or post-embedding reaction? 2. Are you doing immunohistochemistry? If so, what antigen? Pre-embedding or post-embedding reaction? 3. What plastic are you embedding in? 4. What routine stains do you need to do? 5. What part of the brain will you be looking at? The brain is a big place. 6. Do you need to do LM and EM on the same piece of tissue/same cells? Advice on HistoNet is free. Some of it is quite good. Checking published papers in refereed journals and looking at the results is always a good idea, espcially given the cost of experimental animals. Geoff Kristen Reynolds wrote: I need advice on perfusion solutions for rat/monkey brains. I need to use the tissue for both light microscopy and EM. Any suggestions? >Wow, I can't believe how rude of a response I got. I >thought this was for helpful comments. I know how to >perfuse for light microscopy and EM. I just thought >the fixative combination for wanting to do both on the >same tissue might be out there. I guess I won't be >asking histonet anything anymore. > >__________________________________________________ > 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. ------------------------------ Message: 5 Date: Thu, 20 Jan 2005 11:28:47 -0800 From: Rocan Subject: Re: [Histonet] NIH image J surface area unit To: "'histonet@pathology.swmed.edu'" , Subratab Message-ID: <816F081F-6B19-11D9-8D0A-000A9589219E@mac.com> Content-Type: text/plain; charset=US-ASCII; format=flowed I guess there are a couple of ways to do this. I use a stage micrometer (you can google "stage micrometer" and find it on the pictures). I take a picture of the ruler imprinted. Typically these rulers have a one millimeter ruler and 100 divisions of 10 micrometers each. So, with the photo using the same scope and objective you go to any of these programs like NIH ImageJ and open the document (same size as the photos you are measuring). There is a command where you place the mouse on one end drag and click on the other end of the markings. So, if you drag it over ten marking you then instruct the program that that distance is 100 micrometers. Then using these settings you do your measurements and the measurements would then be very precise and will be in micrometers. I do this with a different program but I know you can do it with ImageJ. I hope this helps. Rocio ----- Dr.Rocio Sierra-Honigmann Director Engineered Wound Repair Laboratory Cedars Sinai Research Institute Davis 1091 310-423-1882 Honigmannr@cshs.org On Jan 20, 2005, at 10:02 AM, Subratab wrote: > Dear Histo experts > My friend is measuring glomerular surface area by using Image J > software. He > is in trouble to get the unit of the area calculated by the software, I > mean, if the calculated area is to be expressed in square milimeter or > square micrometer. > He is capturing the figures from microscope with X100 magnification > (oil-emersion lense) and then analyzing the glomerular surface area by > the > software in a computer. > Could anybody please help to solve this. > Sincerely > > Subrata Biswas > University of Campinas > SP, Brazil. > > -- > http://www.histosearch.com/homepages/DrSubrataKumarBiswas.html > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Thu, 20 Jan 2005 12:09:42 -0800 (PST) From: Gareth Davis Subject: Re: [Histonet] Certification requirements To: Fran Lemons , Histonet Message-ID: <20050120200942.71181.qmail@web52708.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Fran, I believe it's based on what state you work in. I know that Florida requires an ASCP certification as well as a Florida license. Tennessee, where I work, does not require it. Gareth Davis Fran Lemons wrote: Has anyone out there seen anything in writing about not being able to work in histology labs without HT certification? Thanks Fran Walker _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' ------------------------------ Message: 7 Date: Thu, 20 Jan 2005 12:14:32 -0800 (PST) From: Gareth Davis Subject: Re: [Histonet] Rudeness on histonet To: "Randolph-Habecker, Julie" , Histonet Message-ID: <20050120201432.90980.qmail@web52710.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Here! Here! to Julie. I have also experienced the rudeness and seen others put down and treated as if they were stupid. It's very discouraging. Gareth "Randolph-Habecker, Julie" wrote: Dear Kristen and others: I too feel that Kristen was treated rudely. I have noticed this problem from time to time on HistoNet and I feel I have to say something. Some people generally assume that because you ask an open question that you are clueless. While you might get better answers by being more specific, the better way to respond to "any suggestions" is to ask if you could provide more details. Just assuming that you and your boss have no idea what you are doing and have ventured into this project naively is not fair. That response was not helpful - it was nothing but rude. Did they suggest a class or a book that they found practically good? No! The response was condescending and pointless. Please ask for clarification before you humiliate a person. And please do not tell someone they deserved to be treated rudely - that is even worse! People write into histonet with questions because there is a wealth of knowledge amongst the subscribers. This is valuable information and constitutes years and years of experience. Keep in mind, all of our experiences have been different. Just because a person might not have experience in one area doesn't mean that they are not very knowledgeable in another. In other words, if you are rude to a person they might not want to share valuable information with you when you ask. Or worse, they might leave the Histonet and deprive all of us of their knowledge! I too asked an open question one time about processing tissue with plastic beads in it and received an incredibly condescending response instructing me on how to design an experiment. I have a Ph.D. for god's sake. I have spent 15 years designing experiments. I also saw a young student publicly filleted for cheating when her instructor encouraged her to send her inquiry out to the Histonet. Kristen, please don't leave the histonet because of one (incredibly) rude response. Take it for what it is - useless! Please hold out for helpful people that actually have information to contribute. Hang in there! 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 Message: 2 Date: Thu, 20 Jan 2005 11:44:27 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] perfusion suggestions follow up To: Kristen Reynolds Cc: histonet@lists.utsouthwestern.edu Message-ID: <41F00A1B.6060101@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Dear Kristen: Your original posting (see below) gave no indication that you were familiar with perfusion for LM or EM. You did not ask for fixative composition, you just asked for "any suggestions". Given the broad scope of your question, the advice you got WAS helpful. The fixative combination for wanting to do both on the same tissue IS out there, but you have to be more specific in your question. The correct fixative combinatio will depend on: 1. Are you doing histochemistry? If so, what reaction? Pre-embedding or post-embedding reaction? 2. Are you doing immunohistochemistry? If so, what antigen? Pre-embedding or post-embedding reaction? 3. What plastic are you embedding in? 4. What routine stains do you need to do? 5. What part of the brain will you be looking at? The brain is a big place. 6. Do you need to do LM and EM on the same piece of tissue/same cells? Advice on HistoNet is free. Some of it is quite good. Checking published papers in refereed journals and looking at the results is always a good idea, espcially given the cost of experimental animals. Geoff Kristen Reynolds wrote: I need advice on perfusion solutions for rat/monkey brains. I need to use the tissue for both light microscopy and EM. Any suggestions? >Wow, I can't believe how rude of a response I got. I >thought this was for helpful comments. I know how to >perfuse for light microscopy and EM. I just thought >the fixative combination for wanting to do both on the >same tissue might be out there. I guess I won't be >asking histonet anything anymore. > >__________________________________________________ > 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 --------------------------------- Do you Yahoo!? Yahoo! Search presents - Jib Jab's 'Second Term' ------------------------------ Message: 8 Date: Thu, 20 Jan 2005 12:49:37 -0800 (PST) From: Dawn Cowie Subject: [Histonet] certification requirements to work in histo labs To: histonet Message-ID: <20050120204937.7153.qmail@web81008.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Fran, In Florida where I am, to perform tech duties you must be licensed. However, you may employ non techs to work as lab aides. These aides may perform a wide variety of duties that usually are done by techs. Specifically what they cannot do is: embed, cut, stain, frozen sections etc. What they can do is: change reagents, coverslip, start and stop an automated stainer. I hope this helps. Dawn Cowie, HT Pensacola Path ------------------------------ Message: 9 Date: Thu, 20 Jan 2005 12:57:54 -0800 From: "Robyn Vazquez" Subject: Re: [Histonet] Rudeness on histonet To: histonet@lists.utsouthwestern.edu, jhabecke@seattlecca.org Message-ID: Content-Type: text/plain; charset=us-ascii BRAVO ladies for speaking your mind!!!! :0) Robyn >>> "Randolph-Habecker, Julie" 1/20/2005 11:27:09 AM >>> Dear Kristen and others: I too feel that Kristen was treated rudely. I have noticed this problem from time to time on HistoNet and I feel I have to say something. Some people generally assume that because you ask an open question that you are clueless. While you might get better answers by being more specific, the better way to respond to "any suggestions" is to ask if you could provide more details. Just assuming that you and your boss have no idea what you are doing and have ventured into this project naively is not fair. That response was not helpful - it was nothing but rude. Did they suggest a class or a book that they found practically good? No! The response was condescending and pointless. Please ask for clarification before you humiliate a person. And please do not tell someone they deserved to be treated rudely - that is even worse! People write into histonet with questions because there is a wealth of knowledge amongst the subscribers. This is valuable information and constitutes years and years of experience. Keep in mind, all of our experiences have been different. Just because a person might not have experience in one area doesn't mean that they are not very knowledgeable in another. In other words, if you are rude to a person they might not want to share valuable information with you when you ask. Or worse, they might leave the Histonet and deprive all of us of their knowledge! I too asked an open question one time about processing tissue with plastic beads in it and received an incredibly condescending response instructing me on how to design an experiment. I have a Ph.D. for god's sake. I have spent 15 years designing experiments. I also saw a young student publicly filleted for cheating when her instructor encouraged her to send her inquiry out to the Histonet. Kristen, please don't leave the histonet because of one (incredibly) rude response. Take it for what it is - useless! Please hold out for helpful people that actually have information to contribute. Hang in there! 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 Message: 2 Date: Thu, 20 Jan 2005 11:44:27 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] perfusion suggestions follow up To: Kristen Reynolds Cc: histonet@lists.utsouthwestern.edu Message-ID: <41F00A1B.6060101@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Dear Kristen: Your original posting (see below) gave no indication that you were familiar with perfusion for LM or EM. You did not ask for fixative composition, you just asked for "any suggestions". Given the broad scope of your question, the advice you got WAS helpful. The fixative combination for wanting to do both on the same tissue IS out there, but you have to be more specific in your question. The correct fixative combinatio will depend on: 1. Are you doing histochemistry? If so, what reaction? Pre-embedding or post-embedding reaction? 2. Are you doing immunohistochemistry? If so, what antigen? Pre-embedding or post-embedding reaction? 3. What plastic are you embedding in? 4. What routine stains do you need to do? 5. What part of the brain will you be looking at? The brain is a big place. 6. Do you need to do LM and EM on the same piece of tissue/same cells? Advice on HistoNet is free. Some of it is quite good. Checking published papers in refereed journals and looking at the results is always a good idea, espcially given the cost of experimental animals. Geoff Kristen Reynolds wrote: I need advice on perfusion solutions for rat/monkey brains. I need to use the tissue for both light microscopy and EM. Any suggestions? >Wow, I can't believe how rude of a response I got. I >thought this was for helpful comments. I know how to >perfuse for light microscopy and EM. I just thought >the fixative combination for wanting to do both on the >same tissue might be out there. I guess I won't be >asking histonet anything anymore. > >__________________________________________________ > 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 ------------------------------ Message: 10 Date: Thu, 20 Jan 2005 12:58:14 -0800 (PST) From: Georgia Stewart Subject: [Histonet] HIPPA rules and fax transmittals going to wrong person To: histonet Message-ID: <20050120205814.70166.qmail@web81402.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hello and Good Afternoon Everyone In this era of HIPPA rules and regulations in the United States, I have not seen any comment on HIPPA as it relates to fax transmittal's of reports that might go to the wrong person/facility. If a report goes to the the wrong person/facility, isn't this is a breach of patient confidentiality covered by HIPPA? How are you handling faxs that go, or might go, to the wrong person/facility? Appreciate your comments. Georgia Stewart, BS, HTL Georgia ------------------------------ Message: 11 Date: Thu, 20 Jan 2005 15:00:39 -0600 From: "Rittman, Barry" Subject: RE: [Histonet] perfusion suggestions To: "histonet" Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0012C92C3@UTHEVS3.mail.uthouston.edu> Content-Type: text/plain; charset="us-ascii" Kristen I am not trying to justify any response that you received but I think that I understand why the response was phrased in that particular manner. Many of the questions that arrive at Histonet are very specific, and generally get a lot of responses. The broader questions are often difficult to answer (and in general I personally do not respond to these) and often receive few responses. It is usually not possible to determine the level of knowledge of the individual who is asking the question and there may be a wide variety of answers. Here on Histonet we are often between a rock and a hard place. I believe that it is a natural tendency nowadays to pose the question in the shortest way possible on the assumption that nobody wishes to spend their time to read a long discourse. This unfortunately often results in a communication that is open to a wide variety of interpretations. One interpretation is that the individual posing the question wants to be informed about all aspects of a particular technique from anesthetizing an animal to interpreting the final result, rather than carrying out the initial groundwork themselves. There is usually no way to judge if this is true from the original communications. This is the era of rapid fire information but unfortunately the recipients do not always have the same background or experiences to view that information in the same light. In many cases I feel that in response to complex questions, individuals with the appropriate expertise should provide their telephone number so that a dialogue can be set up. While Histonet is very useful, a one on one dialogue generally will provide the most relevant information. I hope that you will continue to ask questions on Histonet. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Reynolds Sent: Thursday, January 20, 2005 9:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] perfusion suggestions Wow, I can't believe how rude of a response I got. I thought this was for helpful comments. I know how to perfuse for light microscopy and EM. I just thought the fixative combination for wanting to do both on the same tissue might be out there. I guess I won't be asking histonet anything anymore. __________________________________________________ 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 ------------------------------ Message: 12 Date: Thu, 20 Jan 2005 13:24:08 -0800 From: Victor Tobias Subject: Re: [Histonet] HIPPA rules and fax transmittals going to wrong person To: Georgia Stewart Cc: histonet Message-ID: <41F02178.1080606@pathology.washington.edu> Content-Type: text/plain; charset=us-ascii; format=flowed We had an incident where a patient report went to a Barnes and Noble bookstore. At the time of the incident many users had access to our database for inputting new referring physicians. It is very easy to make a typo and the fax is going who knows where. The bookstore contacted us regarding the incident. We confirmed that we had their fax number in our database. An incident report was filed with risk management. Since the incident, we have restricted access to our database. Our facility recommendation is to verify fax numbers quarterly. With our staffing levels this is not feasible. We do send out annual verifications, that whoever signs the form is stating the fax number is correct and the machine is in a secure location. We will make several attempts to get the verification back. If there is no response then we will stop faxing to them and send a print copy through the mail. We have custom reports created to query the database when a physician is listed on a current case and their fax verification is more than a year old. Our facility policy states that a reasonable effort will be made to verify a fax number. What happens when a doctors office calls and asks for a copy of a patient report? We document in the database who we sent a copy of the report to. Victor Georgia Stewart wrote: >Hello and Good Afternoon Everyone > >In this era of HIPPA rules and regulations in the United States, I have not seen any comment on HIPPA as it relates to fax transmittal's of reports that might go to the wrong person/facility. If a report goes to the the wrong person/facility, isn't this is a breach of patient confidentiality covered by HIPPA? How are you handling faxs that go, or might go, to the wrong person/facility? > >Appreciate your comments. > >Georgia Stewart, BS, HTL > > > >Georgia >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. ------------------------------ Message: 13 Date: Thu, 20 Jan 2005 22:04:33 GMT From: "mprice26@juno.com" Subject: [Histonet] RE: Rudeness on Histonet To: histonet@lists.utsouthwestern.edu Message-ID: <20050120.140511.23783.21286@webmail26.nyc.untd.com> Content-Type: text/plain I have been treated rudely also. I have e-mailed the histonet for a vendor's # or for other info and I will have people rerspond back with try google search. I know how to search for something via google or yellowpages.com but prefer to use the histonet because of the vast number of vendors that subscribe to the histonet. So not only do I receive the # I am asking for I also receive info from other vendors that I would not receive if I look it up on google. I wish those that think I am an idiot for not using google would just not respond to my request. If it bothers them so much that I am requesting info via the Histonet. It takes much less time to hit their delete button than it does for them to e-mail me a rude, condescending message. I would like to add that for the most part I receive prompt useful replys, it is just a few people that reply with rude answers. Marsha Price ------------------------------ Message: 14 Date: Thu, 20 Jan 2005 14:13:17 -0800 (PST) From: pam plumlee Subject: [Histonet] GLP lab staining question To: HistoNet Server Message-ID: <20050120221317.48866.qmail@web11605.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hello fellow histonetters: I'm setting up a special stain control/slide bank for our research pharm lab. I'm curious how special stain controls are validated. Is it as simple as screening the stained slides/blocks as they are cut and determining if they are positive or not, along with the proper documentation? Anyone care to share info? Thanks, PP __________________________________ Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. http://mobile.yahoo.com/maildemo ------------------------------ Message: 15 Date: Thu, 20 Jan 2005 15:14:25 -0700 From: Gayle Callis Subject: Re: [Histonet] certification requirements to work in histo labs To: Dawn Cowie , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050120150521.01b1fbc0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I am curious or rather need clarification here. Licensed means licensed by the state but HT(SCP) is a certification granted by a certification agency, ASCP/CAP. I don't think ASCP/CAP is governed by any state's' licensing regulations. Many moons ago, in New York, one did not have to be certified as an HT to be licensed by the state (one had to take a test for NY licensure), and there were NY licensed technicians without HT(ASCP) certification performing tech duties and, in fact, was the supervisor of our histopathology lab. At 01:49 PM 1/20/2005, you wrote: >Fran, > >In Florida where I am, to perform tech duties you must be licensed. >However, you may employ non techs to work as lab aides. These aides may >perform a wide variety of duties that usually are done by techs. >Specifically what they cannot do is: embed, cut, stain, frozen sections >etc. What they can do is: change reagents, coverslip, start and stop an >automated stainer. I hope this helps. > >Dawn Cowie, HT > >Pensacola Path > >_______________________________________________ >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: 16 Date: Thu, 20 Jan 2005 14:36:24 -0800 From: Bryan Llewellyn Subject: Re: [Histonet] GLP lab staining question To: Histonet Message-ID: <001601c4ff40$7923b6b0$80004246@yourlk4rlmsu> Content-Type: text/plain; reply-type=original; charset=iso-8859-1; format=flowed The way I was taught was to cut a series of sections, 50 or so, then stain the first and the last to make sure they were positive If they both are, it is presumed that all the others are also. If not, stain intermediate sections until one is positive, then use from the first one to that. Bryan Llewellyn ----- Original Message ----- From: "pam plumlee" To: "HistoNet Server" Sent: Thursday, January 20, 2005 2:13 PM Subject: [Histonet] GLP lab staining question > Hello fellow histonetters: I'm setting up a special > stain control/slide bank for our research pharm lab. > I'm curious how special stain controls are validated. > Is it as simple as screening the stained slides/blocks > as they are cut and determining if they are positive > or not, along with the proper documentation? Anyone > care to share info? Thanks, PP > > > > __________________________________ > Do you Yahoo!? > Take Yahoo! Mail with you! Get it on your mobile phone. > http://mobile.yahoo.com/maildemo > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Thu, 20 Jan 2005 16:45:10 -0600 From: "Linda Jones" Subject: [Histonet] Special stain To: Message-ID: Content-Type: text/plain; charset=US-ASCII How do you charge for certain stains? Should charge due to the length of time it take to do the stain or the price of the chemicals? thanks Linda Harper-Jones BS.,HT/ HTL(ASCP) University of Mississippi Medical Center Department of Pathology Chief Histotechnologist Supervisor (601) 984-1576 (601) 984-4968 Fax ljones@pathology.umsmed.edu ------------------------------ Message: 18 Date: Thu, 20 Jan 2005 15:49:45 -0800 From: Jennifer MacDonald Subject: Re: [Histonet] Special stain To: "Linda Jones" Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Special stains are based on the type of stain. For billing purposes there are two types: microorganism or not. 88312 and 88313. Cost or length of time is not a factor when billing. "Linda Jones" Sent by: histonet-bounces@lists.utsouthwestern.edu 01/20/2005 02:45 PM To cc Subject [Histonet] Special stain How do you charge for certain stains? Should charge due to the length of time it take to do the stain or the price of the chemicals? thanks Linda Harper-Jones BS.,HT/ HTL(ASCP) University of Mississippi Medical Center Department of Pathology Chief Histotechnologist Supervisor (601) 984-1576 (601) 984-4968 Fax ljones@pathology.umsmed.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Fri, 21 Jan 2005 04:04:26 +0000 From: Kim O'Sullivan Subject: [Histonet] Bouins fixation problem To: histonet@lists.utsouthwestern.edu Message-ID: <130.194.114.210.1106279796.85555@my.monash.edu.au> Content-Type: text/plain Hi everybody, We have used bouin's fixative for fixing mouse kidneys for years with no problem, and have recently been experiencing problems cutting them once they have been processed and wax embedded. The kidney tissue has holes in it in the medulla region, is very difficult to cut(tissue brittle) and it appears almost like the wax has not infiltrated the kidneys properly. Additonally once the sections have been mounted on glass slides they appear fine but when we go to PAS stain them half of them come off the slides in solution (this has never happened before in the last 3 years)I have checked the processing schedule and that is fine (as is all the solutions) I have checked the temperature of the wax (which is fine). Does anyone out there know what our problem may be?? Additionally is there any way that any of the solutions used to make bouin's (picric acid, formalin and glacial acetic acid)can go off? And how would you check for this. Would appreciate any help!! Kim O'Sullivan ------------------------------ Message: 20 Date: Fri, 21 Jan 2005 05:24:15 -0800 (PST) From: Kim Merriam Subject: [Histonet] c-fos antibody in mouse tissue To: Histonet Message-ID: <20050121132416.20426.qmail@web52502.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hello All, I was wondering if anyone knew what tissue would make the best positive control for this antibody in mouse tissue. Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 21 Date: Fri, 21 Jan 2005 09:23:30 -0500 From: "Angela Bitting" Subject: [Histonet] Coverslipping tape To: Message-ID: Content-Type: text/plain; charset=US-ASCII What is the concensus of opinion on Mercedes Medicals coverslipping tape? I got a sample to try and the girls I work with said they had it before, (I don't know how long ago) and in a couple days it started lifting. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 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: 22 Date: Fri, 21 Jan 2005 09:25:41 EST From: DPALLP@aol.com Subject: [Histonet] saturated fingertips To: histonet@pathology.swmed.edu Message-ID: <191.372a8459.2f226ae5@aol.com> Content-Type: text/plain; charset="US-ASCII" Does anyone have any suggestions to alleviate water saturated fingertips from handling blocks from ice trays? Wearing gloves is too cumbersome and the different barrier lotions that we have tried do not seem to have an effect. Susie ------------------------------ Message: 23 Date: Fri, 21 Jan 2005 09:39:29 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] Bouins fixation problem To: Kim O'Sullivan Cc: histonet@lists.utsouthwestern.edu Message-ID: <41F13E51.1010507@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Hi Kim: Hmm. Certainly sounds like an infiltration problem. My first though is that you are not getting all of the alcohol out of the medulla so xylene (or substitute) is not getting in so the wax is not getting in. When you say that you have checked all of the processing solutions and they are fine, how did you check them? What is the criteria for deciding that a solution is good? I would dump ALL of the processing solutions and ALL of the wax and start fresh. Run some kidneys, cut them, and see if that solves the problem. There is a small chance that the manufacturer of your wax has changed the formula but with modern waxes and additives such a change seems unlikely to cause problems. As for reagents going bad, picric acid last forever, formaldehyde is fine as long as there is no ppt. on the bottom of the bottle, I don't think glacial acetic acid goes bad. Even if the fixation was incomplete (you did not say how long you fix the kidneys, if you perfuse fixative, or if you slice them open) the alcohols should finish the fixation. Good luck! Geoff Kim O'Sullivan wrote: >Hi everybody, > >We have used bouin's fixative for fixing mouse kidneys for years with no problem, and have recently been experiencing problems cutting them once they have been processed and wax embedded. The kidney tissue has holes in it in the medulla region, is very difficult to cut(tissue brittle) and it appears almost like the wax has not infiltrated the kidneys properly. Additonally once the sections have been mounted on glass slides they appear fine but when we go to PAS stain them half of them come off the slides in solution (this has never happened before in the last 3 years)I have checked the processing schedule and that is fine (as is all the solutions) I have checked the temperature of the wax (which is fine). Does anyone out there know what our problem may be?? Additionally is there any way that any of the solutions used to make bouin's (picric acid, formalin and glacial acetic acid)can go off? And how would you check for this. > >Would appreciate any help!! > >Kim O'Sullivan > >_______________________________________________ >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: 24 Date: Fri, 21 Jan 2005 10:00:37 -0500 From: "Rice, Michael" Subject: RE: [Histonet] Coverslipping tape To: "Angela Bitting" , Message-ID: <3BC92F29BE821745AB15E04C98EE028D693742@HCH2KMAIL.holy-cross.com> Content-Type: text/plain I have been using it for several months now and have seen no evidence of lifting, not to say that it wont happen in the future. A friend of mine has beeen using it for several years now with no problems Mike Rice Holy Cross hospital ft lauderdale -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Friday, January 21, 2005 9:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Coverslipping tape What is the concensus of opinion on Mercedes Medicals coverslipping tape? I got a sample to try and the girls I work with said they had it before, (I don't know how long ago) and in a couple days it started lifting. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- ------------------------------ Message: 25 Date: Fri, 21 Jan 2005 09:01:05 -0600 From: "Dawson, Glen" Subject: [Histonet] RE: Rudeness on Histonet To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Rude people on a listserver like this are unavoidable. Unfortunately, there is no "pre-membership rudeness testing". If you are tired of being chewed on like a bloody wildabeast in the midst of a den of lions, do what I was forced to do and avoid lengthy general postings as much as possible. Submit a pithy comment/question and pursue only those who seem to genuinely want to help and delete those who are jerks. Trying to rail against rudeness can become a full time job which doesn't pay anything. Histonet is too valuable a tool to just get rid of, though, at times, it is tempting. Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI ------------------------------ Message: 26 Date: Fri, 21 Jan 2005 10:10:31 EST From: KarBieber@aol.com Subject: [Histonet] Use of Sta-On for Immuno stains To: histonet@lists.utsouthwestern.edu Message-ID: <194.36bbc139.2f227567@aol.com> Content-Type: text/plain; charset="US-ASCII" I was just wondering how other labs are handling this situation: For prostate and breast core biopsies we cut extra slides for potential immuno staining, between the levels. The problem is the use of adhesive in the water bath. Since we are cutting H&E's, there is the possibility of fall off during staining if we DON'T use sta-on. But using it increases the possibility of fall off during immunos staining. And if we don't use the sta-on, we run the risk of fall off on ALL the cases the techs are cutting that day. I know there are lots of labs out there performing immunos, so I'd like to know how you've resolved this. Thanks, Karen ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 14, Issue 26 **************************************** ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From wbedale <@t> biochem.wisc.edu Fri Jan 21 10:32:03 2005 From: wbedale <@t> biochem.wisc.edu (wbedale@biochem.wisc.edu) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Intestinal Tissue Questions Message-ID: <2128.128.104.117.175.1106325123.squirrel@128.104.117.175> Hi to all of you, I'm quite new to histology and have a few basic questions. Here's what I'm doing: Tissue: mouse intestine; small and large intestine is harvested, cut open longitudinally, and rinsed with PBS prior to fixation; 3 cm lengths are fixed. Fixation: 10% formalin Processing and embedding: manual processing, paraffin embedding, 4-6 micron sections Staining: H&E staining Two questions: 1. After fixation, is it better to store my tissue (for ~1 month or so) in fixative, or in 70% ethanol? 2. I'm attempting to get transverse sections of the intestine; sometimes I notice that my tissue ends up inside out (mucosa facing outward). I think it may happen when I use a razor blade to cut the tissue longitudinally vs. using scissors to make the longitudinal incision, and am going to test that. Has anyone had any experience with this? Thanks for any advice. Wendy Wendy Bedale, Ph.D. Assistant Scientist Department of Biochemistry University of Wisconsin-Madison 433 Babcock Dr. Madison, WI 53706 608-262-3099 ext. 3266 wbedale@biochem.wisc.edu From mbecker <@t> pathlabinc.com Fri Jan 21 10:33:08 2005 From: mbecker <@t> pathlabinc.com (Michelle Becker) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] Coverslipping tape In-Reply-To: <3BC92F29BE821745AB15E04C98EE028D693742@HCH2KMAIL.holy-cross.com> Message-ID: We had alot of problems with the Mercedes Medical coverslipping tape. Our Coverslipper did not operate well, even after adjustments were made by their service rep. We also experienced alot of "curling" of the tape. It was better for us to stick with the Sakura tape eventhough it was a little more expensive. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rice, Michael Sent: Friday, January 21, 2005 10:01 AM To: Angela Bitting; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Coverslipping tape I have been using it for several months now and have seen no evidence of lifting, not to say that it wont happen in the future. A friend of mine has beeen using it for several years now with no problems Mike Rice Holy Cross hospital ft lauderdale -----Original Message----- From: Angela Bitting [mailto:akbitting@geisinger.edu] Sent: Friday, January 21, 2005 9:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Coverslipping tape What is the concensus of opinion on Mercedes Medicals coverslipping tape? I got a sample to try and the girls I work with said they had it before, (I don't know how long ago) and in a couple days it started lifting. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ---------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Fri Jan 21 10:35:47 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:24:31 2005 Subject: [Histonet] perfusion suggestions Message-ID: Kristen, Many have replied before, and all wisely. However, rude to one is terse or one of many other descriptions to another. Invariably it will be well meaning. Many of us have been rude, albeit unintentionally, and been rude to. Ask yourself this question when dealing with a conceived rude post - "Was this posted merely to be rude to me?" Unless the answer is unquestionably yes ..... my advice is to learn to ride it (the style). 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: Kristen Reynolds [mailto:kristenhinkle@yahoo.com] Sent: 20 January 2005 15:47 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] perfusion suggestions Wow, I can't believe how rude of a response I got. I thought this was for helpful comments. I know how to perfuse for light microscopy and EM. I just thought the fixative combination for wanting to do both on the same tissue might be out there. I guess I won't be asking histonet anything anymore. __________________________________________________ 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 Fri Jan 21 10:52:06 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Intestinal Tissue Questions Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0012C939D@UTHEVS3.mail.uthouston.edu> Hi Here in the lone star state we generally say y'all to an individual or y'all y'all to a group. Hi to y'all in Wisconsin - we lived in Iowa for several years and enjoyed visits to several wonderful places in your state. 1. My suggestion is that once you have fixed in buffered formalin is to store in 70% ethanol. If for a longer time then you may wish to consider adding 10-20% glycerin. This is expensive but guarantees that tissue will never dry out. Only reason not to use 70% is if you wish to do frozens for lipids some time later as some lipids will be lost. 2. Most mucosae will tend to curl in this manner. What I would recommend is to pin the tissue on a piece of cork and then place tissue side down in the fixative. Must be careful to use stainless steel or plastic pins as some regular steel pins will rust onto the tissue. (we used to use small hedgehog quills for this purpose but these are not readily available here). Once tissue is fixed then it should not curl up and can be processed normally. It does not take long for tissue to become hardened sufficiently to allow you to remove from the cork so that fixation can be completed after removal. If you have really small pieces can place the fresh tissue on a piece of thin card such as postcard with the connective tissue side down for fixing. The tissue fluids will bind the gut to the card. Would recommend removal of card prior to sectioning! Hope this helps Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of wbedale@biochem.wisc.edu Sent: Friday, January 21, 2005 10:32 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Intestinal Tissue Questions Hi to all of you, I'm quite new to histology and have a few basic questions. Here's what I'm doing: Tissue: mouse intestine; small and large intestine is harvested, cut open longitudinally, and rinsed with PBS prior to fixation; 3 cm lengths are fixed. Fixation: 10% formalin Processing and embedding: manual processing, paraffin embedding, 4-6 micron sections Staining: H&E staining Two questions: 1. After fixation, is it better to store my tissue (for ~1 month or so) in fixative, or in 70% ethanol? 2. I'm attempting to get transverse sections of the intestine; sometimes I notice that my tissue ends up inside out (mucosa facing outward). I think it may happen when I use a razor blade to cut the tissue longitudinally vs. using scissors to make the longitudinal incision, and am going to test that. Has anyone had any experience with this? Thanks for any advice. Wendy Wendy Bedale, Ph.D. Assistant Scientist Department of Biochemistry University of Wisconsin-Madison 433 Babcock Dr. Madison, WI 53706 608-262-3099 ext. 3266 wbedale@biochem.wisc.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Fri Jan 21 14:13:09 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Intestinal Tissue Questions In-Reply-To: <2128.128.104.117.175.1106325123.squirrel@128.104.117.175> References: <2128.128.104.117.175.1106325123.squirrel@128.104.117.175> Message-ID: <41F16255.1040308@umdnj.edu> Hi Wendy: wbedale@biochem.wisc.edu wrote: >Hi to all of you, > >I'm quite new to histology and have a few basic questions. > >Here's what I'm doing: > >Tissue: mouse intestine; small and large intestine is harvested, cut >open longitudinally, and rinsed with PBS prior to fixation; 3 cm lengths >are fixed. >Fixation: 10% formalin >Processing and embedding: manual processing, paraffin embedding, 4-6 >micron sections >Staining: H&E staining > >Two questions: > >1. After fixation, is it better to store my tissue (for ~1 month or so) >in fixative, or in 70% ethanol? > How long are you fixing? 48 hours is a minimum, a week is probably better. Formalin works slowly. I don't know exactly what you are looking for but I would try to standardize my processing schedule so all tissues were treated for similar time periods. Storage in 70% is OK, but some cytoplasmic components may be extracted. I would just go ahead and embed rather than store in 70% for more than a week. Someone has certainly studied this, but back in the 1950's, or even earlier. >2. I'm attempting to get transverse sections of the intestine; sometimes >I notice that my tissue ends up inside out (mucosa facing outward). I >think it may happen when I use a razor blade to cut the tissue >longitudinally vs. using scissors to make the longitudinal incision, and >am going to test that. Has anyone had any experience with this? > Smooth muscle (in the muscularis externa) does this. If you want nice cross sections cut the intestine open and pin it out flat on a piece of cork sheet (Home Depot) or dental baseplate wax (see your dentist, much cheaper than a supply house) using insect pins (#2 is a good size) while it is fixing. You can turn the cork/was sheet upside down in a jar of fixative. BETTER: don't cut it lengthwise, use a syringe or a pasteur pipet to wash the food, etc out of the lumen then tie off both ends with suture material/dental floss/thread leaving some fix in the lumen, enough to fill the gut but not stretch it out too much. Make a sausage, so to speak. After fixation, cut off the tied parts and you will have a nice, round piece of gut. Works better on small intestine than on large, more substantial wall in the small gut. Call me on the phone for a method to "jelly roll" the whole gut. Geoff (who worked on the small intesting during the last century) > >Thanks for any advice. > >Wendy > >Wendy Bedale, Ph.D. >Assistant Scientist >Department of Biochemistry >University of Wisconsin-Madison >433 Babcock Dr. >Madison, WI 53706 >608-262-3099 ext. 3266 >wbedale@biochem.wisc.edu > > >_______________________________________________ >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 POWELL_SA <@t> Mercer.edu Fri Jan 21 11:17:25 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Use of Sta-On for Immuno stains In-Reply-To: <194.36bbc139.2f227567@aol.com> Message-ID: In the lab where I do PRN work, they sit a small glass dish of deionized water inside one end of regular waterbath where they use Sta-On. The deionized water gets heated with the regular waterbath and immuno sections can be floated on the deionized water and picked up with positive charged slides. If your waterbath is not large enough to accommodate a small dish in it, set up a second waterbath beside it. Shirley -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of KarBieber@aol.com Sent: Friday, January 21, 2005 10:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Use of Sta-On for Immuno stains I was just wondering how other labs are handling this situation: For prostate and breast core biopsies we cut extra slides for potential immuno staining, between the levels. The problem is the use of adhesive in the water bath. Since we are cutting H&E's, there is the possibility of fall off during staining if we DON'T use sta-on. But using it increases the possibility of fall off during immunos staining. And if we don't use the sta-on, we run the risk of fall off on ALL the cases the techs are cutting that day. I know there are lots of labs out there performing immunos, so I'd like to know how you've resolved this. Thanks, Karen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lu_ze <@t> sbcglobal.net Fri Jan 21 12:21:21 2005 From: lu_ze <@t> sbcglobal.net (Ze Lu) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Re: NIH image J surface area unit Message-ID: <00b801c4ffe6$02186290$1302a8c0@OPTIMUM2> Dear Subrata, It needs to do spatial calibration if you want to measure the actual unit. There is function called "set scale". First you should observe a standard scale with known distance under microscope. Then measure the pixel number of a line with known distance. Using "set scale" function, input actual length, unit, choose "global" option. There is also a plug in that you can download. It is called "microscope scale". So you don't need to calibrate everytime when you start the program. Ze Lu, Ph.D. Senior Scientist Optimum Therapeutics From gcallis <@t> montana.edu Fri Jan 21 12:41:22 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] mouse intestine fixation (long description, two ways) In-Reply-To: <2128.128.104.117.175.1106325123.squirrel@128.104.117.175> References: <2128.128.104.117.175.1106325123.squirrel@128.104.117.175> Message-ID: <6.0.0.22.1.20050121105230.01af3dc0@gemini.msu.montana.edu> Wendy, We have done this two ways: 1) Never open the intestine. Remove whole intestine carefully to prevent holes which leak during rinses. For easier handling of whole intestinal tract, we remove large intestine from small intestine at cecal junction- easier to rinse two pieces. It takes a bit of practice to see where you can start the rinse. In general, we start rinse at narrow opening and rinse toward larger opening. Rinse fecal contents out with PBS - 50 mls, it may take a bit more. Use a 50 ml syringe with a #18 guage needle (dulled so as to not damage internal mucosa) for easy insertion into open end of intestine. Be gentle, do go int too far or tear tissue. After rinse, fill intestine with NBF until it runs out, it should appear sausage like (sorry folks! for the analogy). Immerse NBF filled intestine into 1:20 volume tissue to NBF for desired fixation time. After fixation, slice intestine into lengths. We found NBF filled intestine stays distended by fixative, and prevents lumen from collapsing on itself. This allows for immediate, good fixation of delicate villi damaged by residual gut digestive enzymes. We maintain the "tube", so to speak, in order to not have the problem you are experiencing. The processed lengths fill nicely with paraffin, and remain as a tubular structure. We embed for midsagittal or longitudinal orientation to visualize PP on serosal on outside (serosal surface) and villi on internal mucosal surface. 2) Be sure you open gut on midline away from peritoneal fascia attachments. This can be done with a tiny straight dissection scissors, called iris scissors, or a very sharp, fine tipped cuticle scissors from WalMart. Lay your opened strips on lengths of filter paper (buy large sheets, cut into strips, get smooth paper) , wet strips with PBS before putting unfixed tissue on paper, prevents sticking. You can manipulate gut a bit so it sticks, with villi facing up, then fold paperstrip like an accordian, immerse into NBF. If you work in a hood, then wet the paper with NBF, the serosal surface will fix a bit, and you won't leave gut sticking to paper when you embed. The fixed intestine will be perfectly flat. Process folded paper with gut inside a cassette, process but remove paper gently at embedding. If all you do is H&E, you can store in NBF. Good luck, you wrote: >Hi to all of you, > >I'm quite new to histology and have a few basic questions. > >Here's what I'm doing: > >Tissue: mouse intestine; small and large intestine is harvested, cut >open longitudinally, and rinsed with PBS prior to fixation; 3 cm lengths >are fixed. >Fixation: 10% formalin >Processing and embedding: manual processing, paraffin embedding, 4-6 >micron sections >Staining: H&E staining > >Two questions: > >1. After fixation, is it better to store my tissue (for ~1 month or so) >in fixative, or in 70% ethanol? > >2. I'm attempting to get transverse sections of the intestine; sometimes >I notice that my tissue ends up inside out (mucosa facing outward). I >think it may happen when I use a razor blade to cut the tissue >longitudinally vs. using scissors to make the longitudinal incision, and >am going to test that. Has anyone had any experience with this? > >Thanks for any advice. > >Wendy Bedale, Ph.D. 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 ploykasek <@t> phenopath.com Fri Jan 21 13:14:23 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] ISH control probe Message-ID: I have a question for those folks doing ISH. What type of probe are you using to determine that the DNA in the tissue is available for hybridization? Thanks for the help. Patti Loykasek PhenoPath Laboratories Seattle, WA From cfavara <@t> niaid.nih.gov Fri Jan 21 13:15:41 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Intestinal Tissue Questions Message-ID: Wendy, You have gotten fabulous advice. I only add what I do. I perfuse the mouse as we are studying brain and this is far superior. I leave the gut intact in the animal. Cut just beneath the stomach, and at the cecal junction, insert a dulled needle per Gayle Callis rinse jelly roll and put on lens paper. Do the same for the lower intestines. Can do all in one but I find this easier. Get some normal mice try everything and practice, practice, practice..pretty soon you will be giving advice. Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: wbedale@biochem.wisc.edu [mailto:wbedale@biochem.wisc.edu] Sent: Friday, January 21, 2005 9:32 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Intestinal Tissue Questions Hi to all of you, I'm quite new to histology and have a few basic questions. Here's what I'm doing: Tissue: mouse intestine; small and large intestine is harvested, cut open longitudinally, and rinsed with PBS prior to fixation; 3 cm lengths are fixed. Fixation: 10% formalin Processing and embedding: manual processing, paraffin embedding, 4-6 micron sections Staining: H&E staining Two questions: 1. After fixation, is it better to store my tissue (for ~1 month or so) in fixative, or in 70% ethanol? 2. I'm attempting to get transverse sections of the intestine; sometimes I notice that my tissue ends up inside out (mucosa facing outward). I think it may happen when I use a razor blade to cut the tissue longitudinally vs. using scissors to make the longitudinal incision, and am going to test that. Has anyone had any experience with this? Thanks for any advice. Wendy Wendy Bedale, Ph.D. Assistant Scientist Department of Biochemistry University of Wisconsin-Madison 433 Babcock Dr. Madison, WI 53706 608-262-3099 ext. 3266 wbedale@biochem.wisc.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Daydawning <@t> wideopenwest.com Fri Jan 21 13:49:50 2005 From: Daydawning <@t> wideopenwest.com (Dawn Truscott) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] rudeness Message-ID: <001801c4fff2$5ee5bf20$6401a8c0@wowway.com> I have not been personally subjected to the histonet rudeness, but have read many reponses from others who have a "holier than thou" attitude. Even though I have 28 yrs of expereince in Histology I now abstain from commenting because I am in sales. I have many times given this resource to my customers to turn to for help. I would hate for any of them to be treated rudely after telling them what a great place it is to go for help. I guessing if you think the question is beneath you, don't bother typing a reply. As mom always said "If you don't have anything nice to say, don't say anything at all". Dawn M. Truscott Recycling New Year's resolutions since 1987. From bmcmahill <@t> incytepathology.com Fri Jan 21 14:07:14 2005 From: bmcmahill <@t> incytepathology.com (Bonnie J. McMahill) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] ISH control probe Message-ID: Patti, We use a "general" human DNA or RNA probe. DAKO has one (X1414) for DNA. Bonnie McMahill Histology Supervisor InCyte Pathology Spokane, WA > -----Original Message----- > From: Patti Loykasek [SMTP:ploykasek@phenopath.com] > Sent: Friday, January 21, 2005 11:14 AM > To: histonet > Subject: [Histonet] ISH control probe > > I have a question for those folks doing ISH. What type of probe are you > using to determine that the DNA in the tissue is available for > hybridization? Thanks for the help. > > Patti Loykasek > PhenoPath Laboratories > Seattle, WA > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ljb <@t> medicine.wisc.edu Fri Jan 21 14:25:26 2005 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] rudeness Message-ID: I very happily sent someone a protocol of mine and then got several messages from others who wanted to inform me that my ideas were all misguided. It felt as though I'd been slapped in the face. Since then, I have refrained from offering any advice to anyone else. Some folks have put themselves on dangerously high pedestals, leaving the rest of to gaze upward to a nasty view! Cindy B >>> Dawn Truscott 1/21/2005 1:49:50 PM >>> I have not been personally subjected to the histonet rudeness, but have read many reponses from others who have a "holier than thou" attitude. Even though I have 28 yrs of expereince in Histology I now abstain from commenting because I am in sales. I have many times given this resource to my customers to turn to for help. I would hate for any of them to be treated rudely after telling them what a great place it is to go for help. I guessing if you think the question is beneath you, don't bother typing a reply. As mom always said "If you don't have anything nice to say, don't say anything at all". Dawn M. Truscott Recycling New Year's resolutions since 1987. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gentras <@t> vetmed.auburn.edu Fri Jan 21 14:34:03 2005 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:24:32 2005 Subject: Fwd: { SPAM 2 }::Re: [Histonet] rudeness Message-ID: <6.0.1.1.0.20050121143342.01b72ec8@mailhost.vetmed.auburn.edu> Amen! I second that motion. Atoska >I guessing if you think the question is beneath you, don't bother >typing a reply. As mom always said "If you don't have anything nice to >say, don't say anything at all". > > >Dawn M. Truscott From gcallis <@t> montana.edu Fri Jan 21 15:13:49 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] A nice reply for TGIF! RE: Intestinal Tissue Questions In-Reply-To: References: Message-ID: <6.0.0.22.1.20050121141200.01b34cd8@gemini.msu.montana.edu> Dear Cynthia, Great message and enough to lift the spirits of all Histonetters!!! It made my day. Have a good weekend everyone! 12:15 PM 1/21/2005, you wrote: >Wendy, > >You have gotten fabulous advice. I only add what I do. I perfuse the mouse >as we are studying brain and this is far superior. I leave the gut intact in >the animal. Cut just beneath the stomach, and at the cecal junction, insert >a dulled needle per Gayle Callis rinse jelly roll and put on lens paper. Do >the same for the lower intestines. Can do all in one but I find this easier. >Get some normal mice try everything and practice, practice, practice..pretty >soon you will be giving advice. > >Cynthia Favara >NIAID/NIH/RML/LPVD >903 South 4th Street >Hamilton, MT 59840 >406-363-9317 > >Disclaimer: >The information in this e-mail and any of its attachments is confidential >and may contain sensitive information. It should not be used by anyone who >is not the original intended recipient. If you have received this e-mail in >error please inform the sender and delete it from your mailbox or any other >storage devices. National Institute of Allergy and Infectious Diseases shall >not accept liability for any statements made that are sender's own and not >expressly made on behalf of the NIAID by one of its representatives > > > >-----Original Message----- >From: wbedale@biochem.wisc.edu [mailto:wbedale@biochem.wisc.edu] >Sent: Friday, January 21, 2005 9:32 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Intestinal Tissue Questions > >Hi to all of you, > >I'm quite new to histology and have a few basic questions. > >Here's what I'm doing: > >Tissue: mouse intestine; small and large intestine is harvested, cut open >longitudinally, and rinsed with PBS prior to fixation; 3 cm lengths are >fixed. >Fixation: 10% formalin >Processing and embedding: manual processing, paraffin embedding, 4-6 micron >sections >Staining: H&E staining > >Two questions: > >1. After fixation, is it better to store my tissue (for ~1 month or so) in >fixative, or in 70% ethanol? > >2. I'm attempting to get transverse sections of the intestine; sometimes I >notice that my tissue ends up inside out (mucosa facing outward). I think >it may happen when I use a razor blade to cut the tissue longitudinally vs. >using scissors to make the longitudinal incision, and am going to test that. >Has anyone had any experience with this? > >Thanks for any advice. > >Wendy > >Wendy Bedale, Ph.D. >Assistant Scientist >Department of Biochemistry >University of Wisconsin-Madison >433 Babcock Dr. >Madison, WI 53706 >608-262-3099 ext. 3266 >wbedale@biochem.wisc.edu > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DOtt <@t> lifecell.com Fri Jan 21 15:23:21 2005 From: DOtt <@t> lifecell.com (Diane Ott) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Unsubscribe Message-ID: <913CC534DD5B7D458B5FABB4322160F6292C60@exch1.lifecell.biz> Please unsubscribe. Thank you. From Barry.R.Rittman <@t> uth.tmc.edu Fri Jan 21 15:53:08 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] rudeness Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0012C941F@UTHEVS3.mail.uthouston.edu> When advice is given it is not necessary for it to be accepted. This is what we all say but if I send something that I know from my experience works then it is a bit "off......" if this is ignored or ridiculed. I feel that in many cases we are comparing chalk and cheese. If you send out a request for determining which is the best hematoxylin you will probably get 20 responses, many of them different. They may all be correct for that particular application in that particular location. Even if an individual states, "I have been using this for 20 years and it has always worked well for me and my pathologist", it does not mean that it will be true for everyone. The response does not tell us what the particular requirements of either the tech or the pathologist are. Some pathologists may be happy with a wide range of results, some with sections that I would consider overstained with eosin and so on. If you feel that a technique that is put forward has some problems you can of respond on Histonet, just keep quiet or call the individual. Many of us keep quiet because of the flack we usually get. In many ways this is a disservice to those that are less experienced in that particular field. If only one technique is out there and we feel that it is in error it is our responsibility to point this out. This can be done by contacting the author or being diplomatic. "In my experience", or "I have found that" are reasonable ways to bring a different point of view to the Histonet audience. The audience will then have more than one point of view and can make up their own minds. Dawn, I appreciate where you are coming from. I attended a presentation on processing of bone that I and a lot of the audience who were experienced in bone work felt had so many errors that it would be a problem if the methods presented were used. No one said a word. For my part and to prevent any embarrassment to the presenter I wrote a one and a half page detailed critique. I also signed it (perhaps a mistake). It did result in a lot of crap for me but did get the message across to the author. I look back and realize that while it spared the presenter from any embarrassment, it did nothing for the less experienced in the audience, many of whom had never been exposed to processing bone. They were only exposed to one point of view. If I had to do this again, my preference would be to yank the presenter off the stage with a shepherds crook - failing that to be diplomatic and bring up the major points during the presentation, for benefit of the audience. The emphasis I place here is on the word diplomatic, people can be constructively critical in a nice way. I hope that y'all have a great weekend. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of LaCinda Burchell Sent: Friday, January 21, 2005 2:25 PM To: Histonet@lists.utsouthwestern.edu; Daydawning@wideopenwest.com Subject: Re: [Histonet] rudeness I very happily sent someone a protocol of mine and then got several messages from others who wanted to inform me that my ideas were all misguided. It felt as though I'd been slapped in the face. Since then, I have refrained from offering any advice to anyone else. Some folks have put themselves on dangerously high pedestals, leaving the rest of to gaze upward to a nasty view! Cindy B >>> Dawn Truscott 1/21/2005 1:49:50 PM >>> I have not been personally subjected to the histonet rudeness, but have read many reponses from others who have a "holier than thou" attitude. Even though I have 28 yrs of expereince in Histology I now abstain from commenting because I am in sales. I have many times given this resource to my customers to turn to for help. I would hate for any of them to be treated rudely after telling them what a great place it is to go for help. I guessing if you think the question is beneath you, don't bother typing a reply. As mom always said "If you don't have anything nice to say, don't say anything at all". Dawn M. Truscott Recycling New Year's resolutions since 1987. _______________________________________________ Histonet mailing list Histonet@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 histology.bc <@t> shaw.ca Fri Jan 21 16:10:58 2005 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Perceived Rudeness on Histonet ... Message-ID: <41F17DF2.2090502@shaw.ca> I have not been paying attention to the Histonet for the past couple of days and so my comments are a little late. The initial inquiry about "advice on perfusion solutions" was a little vague. My own interpretation of this message was that the writer had minimal understanding of the topic and needed comprehensive advice starting at a very basic level. (having read later responses, this is apparently not the case) I am sure John Kiernan's response was not intended to the rude or condescending. I have known John for many years and he is one of the most "giving" individuals on the Histonet. One has only to look through the archives to see the vast number of times that he has provided responses to complex and/or obscure inquiries. He is probably one of Histonet's most patient and valued contributors. My personal interpretation of his response was that he was giving Kristen sound advice so that she would not be drifting vaguely through unknown territory. He was providing "fatherly" advice so that she could go to her boss with a list of steps to be followed to obtain the information she was apparently seeking. There are many enquiries on Histonet where it seems that the writer has minimal understanding of the topic, or has little knowledge of basic concepts. A vaguely worded enquiry may prompt a reply of "buy a book", "go read the book", or "take a course". These comments are not meant to be rude or condescending; sometimes there is just not enough time or space to provide reams of basic information. The Histonet is an invaluable resource, with numerous highly talented, knowledgeable contributors, but it is not the ideal forum for fundamental instruction. Sometimes, people have to go out there and do what John (and many others) did ... read, take courses, experiment with different procedures ... don't expect to be spoon-fed. Just my thoughts, now I have to go shovel snow off the driveway, Paul Bradbury Kamloops, BC Canada From mcauliff <@t> umdnj.edu Fri Jan 21 19:19:32 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] rudeness In-Reply-To: <566FB0B522443D43AF02D2ADBE35A6F0012C941F@UTHEVS3.mail.uthouston.edu> References: <566FB0B522443D43AF02D2ADBE35A6F0012C941F@UTHEVS3.mail.uthouston.edu> Message-ID: <41F1AA24.2040703@umdnj.edu> Continuing to flog this thread ........ I saw only two responses to the original question, apparently both were deemed rude. Whether or not they were helpful seems to be beside the point. Did any of the many offended parties offer answers? Geoff Rittman, Barry wrote: >When advice is given it is not necessary for it to be accepted. This is >what we all say but if I send something that I know from my experience >works then it is a bit "off......" if this is ignored or ridiculed. >I feel that in many cases we are comparing chalk and cheese. >If you send out a request for determining which is the best hematoxylin >you will probably get 20 responses, many of them different. They may all >be correct for that particular application in that particular location. >Even if an individual states, "I have been using this for 20 years and >it has always worked well for me and my pathologist", it does not mean >that it will be true for everyone. The response does not tell us what >the particular requirements of either the tech or the pathologist are. >Some pathologists may be happy with a wide range of results, some with >sections that I would consider overstained with eosin and so on. >If you feel that a technique that is put forward has some problems you >can of respond on Histonet, just keep quiet or call the individual. >Many of us keep quiet because of the flack we usually get. In many ways >this is a disservice to those that are less experienced in that >particular field. If only one technique is out there and we feel that it >is in error it is our responsibility to point this out. This can be done >by contacting the author or being diplomatic. "In my experience", or "I >have found that" are reasonable ways to bring a different point of view >to the Histonet audience. >The audience will then have more than one point of view and can make up >their own minds. >Dawn, I appreciate where you are coming from. I attended a presentation >on processing of bone that I and a lot of the audience who were >experienced in bone work felt had so many errors that it would be a >problem if the methods presented were used. No one said a word. For my >part and to prevent any embarrassment to the presenter I wrote a one and >a half page detailed critique. I also signed it (perhaps a mistake). It >did result in a lot of crap for me but did get the message across to the >author. I look back and realize that while it spared the presenter from >any embarrassment, it did nothing for the less experienced in the >audience, many of whom had never been exposed to processing bone. They >were only exposed to one point of view. If I had to do this again, my >preference would be to yank the presenter off the stage with a shepherds >crook - failing that to be diplomatic and bring up the major points >during the presentation, for benefit of the audience. >The emphasis I place here is on the word diplomatic, people can be >constructively critical in a nice way. >I hope that y'all have a great weekend. >Barry > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of LaCinda >Burchell >Sent: Friday, January 21, 2005 2:25 PM >To: Histonet@lists.utsouthwestern.edu; Daydawning@wideopenwest.com >Subject: Re: [Histonet] rudeness > >I very happily sent someone a protocol of mine and then got several >messages from others who wanted to inform me that my ideas were all >misguided. It felt as though I'd been slapped in the face. Since then, >I have refrained from offering any advice to anyone else. Some folks >have put themselves on dangerously high pedestals, leaving the rest of >to gaze upward to a nasty view! Cindy B > > > >>>>Dawn Truscott 1/21/2005 1:49:50 PM >>>> >>>> >>>> >I have not been personally subjected to the histonet rudeness, but have >read many reponses from others who have a "holier than thou" attitude. >Even though I have 28 yrs of expereince in Histology I now abstain from >commenting because I am in sales. > >I have many times given this resource to my customers to turn to for >help. I would hate for any of them to be treated rudely after telling >them what a great place it is to go for help. > >I guessing if you think the question is beneath you, don't bother >typing a reply. As mom always said "If you don't have anything nice to >say, don't say anything at all". > > >Dawn M. Truscott > >Recycling New Year's resolutions since 1987. >_______________________________________________ >Histonet mailing list >Histonet@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 TJasper <@t> smdc.org Fri Jan 21 16:25:24 2005 From: TJasper <@t> smdc.org (Jasper, Thomas G.) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Rudeness on the Histonet Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E4505F@harrier> Dear All, I have been following this thread with interest and am not surprised that it has finally come to this much discussion. I must concur with Dawn Truscott's comment about not saying anything. Despite what some people might think there really are no dumb questions. The only thing that's dumb is not asking questions. There is a tremendous amount skill, expertise and knowledge on this listserver, yet we are all human and less than perfect. I also understand the reluctance that people could feel after some of the "high horse" responses that have come out on the Histonet. A certain individual that posts frequently on the Histonet seems to be uniquely skilled at delivering thinly veiled derogatory responses. I find this to be tactless and totally unnecessary. It's also a downright shame as many people have sought out this person as a resource only to be rebuffed and chastised. I recall an episode back in 2001 which so surprised me that I printed out the thread and saved it. I then privately e-mailed the party asking for help, a reassuring message that the question posted was valid and deserving of a well mannered response. I realize people are going to say and do what they want to and they should. If someone's post irritates another so much, that a snide remark or some less than kind response is deemed necessary, I would suggest e-mailing these remarks privately. When it comes to answering questions and sharing knowledge, most certainly this information should be out in the public forum for all. The condescension and elitism don't need to be on the Histonet. Thank you. Thomas Jasper HT(ASCP)BAS Anatomic Pathology Coordinator SMDC Clinical Laboratory Duluth, MN tjasper@smdc.org This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients named above. If you are not the intended recipient, you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. As required by federal and state laws, you need to hold this information as privileged and confidential. If you have received this communication in error, please notify the sender and destroy all copies of this communication and any attachments. From immrstambo <@t> hotmail.com Fri Jan 21 19:16:23 2005 From: immrstambo <@t> hotmail.com (Christine Tambasco) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] LEICA ASP300 In-Reply-To: Message-ID: i have had the leica asp300 for at least 2 years and have not experienced any cleaning or processing problems that werent tech errors. I love it! Christine Tambasco HT (ASCP), St Marys Hosptial, Amsterdam, New York 12010, ph-5188417287 >From: "Erin Herter" >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] LEICA ASP300 >Date: Wed, 19 Jan 2005 11:38:02 -0600 > >Hi again all histonetters..... >Making a plea again to anyone (else) who has a Leica ASP300. Have you >had problems and if so how were they resolved? Our processor fails >routinely in the standard cleaning cycle and also during the routine >overnight processing. The service company is good, but we are now out >of warranty. (We are working on getting that extended because it has >been a real "lemon") >Thanks, >Erin Herter >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMacDonald <@t> mtsac.edu Sat Jan 22 00:37:04 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Looking for Crystals Message-ID: Uric acid crystals (urates) should be fixed in absolute alcohol.& nbsp; You want to preserve them, but they are soluble in aqueous solutions. on the proc Jennifer MacDonald -----his To: "Histonet \(E-mail\)" < From: "Paula Lucas" sbcglobal.net Sat Jan 22 10:00:55 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Certification requirements References: Message-ID: <011a01c5009b$8e953080$dea5ff44@domainnotset.invalid> Don't know exactly where you work, or who is saying this, so I'll cover it from a couple of angles: As several other individuals as pointed out, only Florida has a law that says histotechs must be certified (as far as I'm aware, also). It used to be that the lab techs had to take a Florida exam, but that was changed about a year or two ago, and now the ASCP exam is accepted. That said, I also want to add that each institution and/or lab can also set their own standards. CLIA '88 has basically ignored histotechs, so if techs are doing routine histology (i.e., no ISH for example which is high complexity), then they are low complexity, which means just a high school education but with training. But many labs set up their own standards - HS degree - some college - some college bio-chem course - associate degree with bio-chem courses - not HT certified - HT certified - HT certification eligible - must take & pass certifiication exam within X # years of becoming eligible - etc. Same with HTL, with BA/BS degree. If the institution says must take the HT ASCP exam, and the new ASCP requirements are that the candidate must have 60 college credit hours minimum with 12 hours of bio/chem, then that becomes the standard for those taking the certification exam. If you give us a little more information about the situation, we can provide a little more information specific to your query. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48037 ----- Original Message ----- From: "Fran Lemons" To: ; Sent: Thursday, January 20, 2005 1:26 PM Subject: [Histonet] Certification requirements Has anyone out there seen anything in writing about not being able to work in histology labs without HT certification? Thanks Fran Walker _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Sat Jan 22 10:19:47 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Rudeness Message-ID: <012001c5009e$31a42180$dea5ff44@domainnotset.invalid> Two additional comments: 1. Interpretation - Histonet is a WRITTEN format, but when we read it, we put our own inflections to the words. So sometimes I have read a response, and think the person is being rude, and later when I re-read it (or have more information), I realize that I had put the wrong spin on it. Conversely, I have read responses that I thought were helpful, and was later surprised to see others "attacking" the response for being rude. 2. Additional information needed in the original question - I think this is more true of "first time askers" - but those questions that often get the "rude" responses are often the one sentence questions. "I need a schedule to process tissue X." I read this, and don't even know where to start to respond, because I don't know the background on this. - biopsy or 5x5x2 mm? - by hand or which processor? - which solutions? - which fixative? - has the person been a histotech for 20 years and this is the first time for this tissue, or is this person in the second week of a college histology class and doesn't even know how to pronounce "xylene"? This is where I see the comments - "read a book". So - for those asking the questions - please give us a little more background information, so we know exactly where to help you. - What exactly you want - What you have available - What you've tried so far. - Maybe even your certification and maybe your lab (or a hint as to type (hospital, research, private, histology, IHC, etc.) if you don't want to give out the name of your institution. For those willing to help - feel free to politely ask questions for clarification. For those not willing to take the time to ask the questions for clarification, that's OK too. Just don't answer. Answering on Histonet is optional, and a privilege. Just my opinions. Peggy A. Wenk, HTL(ASCP)SLS From lpwenk <@t> sbcglobal.net Sat Jan 22 10:30:11 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Special stain References: Message-ID: <014301c5009f$a5a54900$dea5ff44@domainnotset.invalid> Charges for specials stains relate more to HOW LONG (time) a pathologist needs to diagnose a slide. Microorganisms take longer to find/diagnosis than non-microorganisms stains. So a PAS done for glycogen or mucin is one charge (88312), while a PAS done for fungus is a different charge (88313). Yet, it is the same PAS stain and it costs the same for histology reagents/tech time. Conversely, a GMS done for fungus is the same charge as a PAS done for fungus (88313). Yet GMS costs more - cost of silver, cost of silver disposal, longer tech time, etc. Same with looking for helicobacter - lab cost of Steiner vs. Diff-Quik, but the same reimbursement charge. Pathologists are often not aware of the COST of the stain (reagents, disposal, tech time) in comparison to the REIMBURSEMENT (difference which equals PROFIT or LOSS). When we do a cost analysis, we can often persuade the pathologists to switch to a cheaper stain. Who does the billing and the determination of which stains go into which categories at your institution? There are also different charges if the tissue needed a frozen section, to be decalcified, is an enzyme procedure, etc. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Linda Jones" To: Sent: Thursday, January 20, 2005 5:45 PM Subject: [Histonet] Special stain > How do you charge for certain stains? Should charge due to the length of > time it take to do the stain or the price of the chemicals? > thanks > > > Linda Harper-Jones BS.,HT/ HTL(ASCP) > University of Mississippi Medical Center > Department of Pathology > Chief Histotechnologist Supervisor > (601) 984-1576 > (601) 984-4968 Fax > ljones@pathology.umsmed.edu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Sat Jan 22 10:33:44 2005 From: lpwenk <@t> sbcglobal.net (lpwenk@sbcglobal.net) Date: Fri Sep 16 15:24:32 2005 Subject: [BULK] - [Histonet] saturated fingertips References: Message-ID: <015001c500a0$24469520$dea5ff44@domainnotset.invalid> Partially unwrap a gauze square. Place the ice cube in the middle, and wrap the gauze around the cube, pulling the extra material into a "tail". Now you have a "handle" to hold onto the ice cube, without holding onto the ice. So your fingers don't get as cold, the ice cube doesn't melt as quickly, your fingers don't turn into prunes, the ice lasts longer, etc. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48037 ----- Original Message ----- From: "Fred Underwood" To: ; Sent: Friday, January 21, 2005 10:49 AM Subject: Re: [BULK] - [Histonet] saturated fingertips > Finger cots might work well. > > Fred > > >>> 01/21/05 09:25AM >>> > Does anyone have any suggestions to alleviate water saturated > fingertips > from handling blocks from ice trays? Wearing gloves is too > cumbersome and the > different barrier lotions that we have tried do not seem to have an > effect. > > Susie > _______________________________________________ > Histonet mailing list > Histonet@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 stewart_georgia <@t> sbcglobal.net Sat Jan 22 10:57:05 2005 From: stewart_georgia <@t> sbcglobal.net (Georgia Stewart) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Certification requirements In-Reply-To: <011a01c5009b$8e953080$dea5ff44@domainnotset.invalid> Message-ID: <20050122165705.47041.qmail@web81405.mail.yahoo.com> Good morning I work in California which does not require certification to work in histology labs but I'm unsure how CLIA classifies performing IHC. Is IHC also high complexity? What education/certification is required by CLIA '88 for techs performing high complexity stains? Thanks in advance Georgia Stewart lpwenk@sbcglobal.net wrote: Don't know exactly where you work, or who is saying this, so I'll cover it from a couple of angles: As several other individuals as pointed out, only Florida has a law that says histotechs must be certified (as far as I'm aware, also). It used to be that the lab techs had to take a Florida exam, but that was changed about a year or two ago, and now the ASCP exam is accepted. That said, I also want to add that each institution and/or lab can also set their own standards. CLIA '88 has basically ignored histotechs, so if techs are doing routine histology (i.e., no ISH for example which is high complexity), then they are low complexity, which means just a high school education but with training. But many labs set up their own standards - HS degree - some college - some college bio-chem course - associate degree with bio-chem courses - not HT certified - HT certified - HT certification eligible - must take & pass certifiication exam within X # years of becoming eligible - etc. Same with HTL, with BA/BS degree. If the institution says must take the HT ASCP exam, and the new ASCP requirements are that the candidate must have 60 college credit hours minimum with 12 hours of bio/chem, then that becomes the standard for those taking the certification exam. If you give us a little more information about the situation, we can provide a little more information specific to your query. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48037 ----- Original Message ----- From: "Fran Lemons" To: ; Sent: Thursday, January 20, 2005 1:26 PM Subject: [Histonet] Certification requirements Has anyone out there seen anything in writing about not being able to work in histology labs without HT certification? Thanks Fran Walker _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Georgia From contact <@t> excaliburpathology.com Sat Jan 22 12:21:17 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Perceived rudeness Message-ID: <20050122182118.69540.qmail@web50306.mail.yahoo.com> Bravo Paul Bradbury. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 631 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From jgjulander <@t> cc.usu.edu Sat Jan 22 14:35:16 2005 From: jgjulander <@t> cc.usu.edu (Justin Julander) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] integrin IHC Message-ID: <41F2BE12@webster.usu.edu> I want to do immunohistochemistry on cell culture for integrin alphaV and beta3. Does anyone here have experience staining for this integrin? I have a peroxidase conjugated secondary Ab, and also am looking for a good basic protocol using that kind of 2?. Thanks, Justin J 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 asmith <@t> mail.barry.edu Sat Jan 22 14:41:53 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] calling all muscle folks... PHOS coverslipping Message-ID: <4C051EAE581BB646BF53A749A73FBA2D1F3C81@exchsrv01.barrynet.barry.edu> In their original paper, Takeuchi and Kuriaki pointed out that one could preserve the stain by adding iodine to the alcohols, xylene, and mounting resin. Haruo Machida, Edwin Perkins, and I took the trouble to find the ideal iodine concentrations: 1 mg/ml of alcohol, xylene, or mounting resin. We found that the best resin for this purpose was Clay-Adams' "Histoclad". Harleco's HSR was the only other satisfactory mounting resin. (Durable mounts of the iodine stain for the phosphorylase reaction, Stain Technology 41: 346-347, 1966) I was able to take publishable photographs of amylo-phosphorylase reactions preserved this way 25 years after the slides were made. 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 Jennifer Hofecker Sent: Friday, January 07, 2005 12:22 PM To: Celebre Julia Cc: histonet posts Subject: RE: [Histonet] calling all muscle folks... PHOS coverslipping Hi Julia, after applying the iodine and air drying, we use 85% Karo syrup to put a coverslip on the slide. It stays readable for about a weeek, but never gets solid and of course, is not considered permanent. We have a big gooey mess of PHOS slides in a drawer that are unreadable. I may try some glycerine. Thanks for responding, Jennifer --- Celebre Julia wrote: > Hi there... > Just curious, when you do coverslip your > Phosphorylases is this permanent? > I personally don't coverslip mine, I grade them for > the pathologist. But one > of my co-workers would use a few drops of iodine > with glycerine and place a > coverslip on top... > > Julia Celebre MLT > Anatomic Pathology > Hamilton General Hospital > 905-527-0271 ext46145 > email: celebrej@hhsc.ca > > > -----Original Message----- > From: Jennifer Hofecker [mailto:jhofecker@yahoo.com] > Sent: Friday, January 07, 2005 11:50 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] calling all muscle folks... PHOS > coverslipping > > > Hello to all muscle pros. > I was wondering what you are using to coverslip your > amylo-phosphorylase (PHOS) slides? Any alternatives > to 85% Karo syrup? > Thanks in advance, > Jennifer > > > > __________________________________ > Do you Yahoo!? > Yahoo! Mail - Easier than ever with enhanced search. > Learn more. > http://info.mail.yahoo.com/mail_250 > > _______________________________________________ > 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. > > > __________________________________ Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. http://promotions.yahoo.com/new_mail _______________________________________________ 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 subratab <@t> bdonline.com Sun Jan 23 13:00:56 2005 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Re: NIH image J surface area unit Message-ID: <200501231903.j0NJ3pYb032315@mailout.proshikanet.com> Dear Ze Lu thank you very much to all of you replyed to my question. Subrata. -- http://www.histosearch.com/homepages/DrSubrataKumarBiswas.html Message: 1 Date: Fri, 21 Jan 2005 13:21:21 -0500 From: "Ze Lu" Subject: [Histonet] Re: NIH image J surface area unit To: Message-ID: <00b801c4ffe6$02186290$1302a8c0@OPTIMUM2> Content-Type: text/plain; charset="iso-8859-1" Dear Subrata, It needs to do spatial calibration if you want to measure the actual unit. There is function called "set scale". First you should observe a standard scale with known distance under microscope. Then measure the pixel number of a line with known distance. Using "set scale" function, input actual length, unit, choose "global" option. There is also a plug in that you can download. It is called "microscope scale". So you don't need to calibrate everytime when you start the program. Ze Lu, Ph.D. Senior Scientist Optimum Therapeutics From greddy9734 <@t> hotmail.com Sun Jan 23 17:26:19 2005 From: greddy9734 <@t> hotmail.com (goutham reddy) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] I was looking for Ki-67 antibody for immunohistochemistry from NovoCastra labs Message-ID: I can't seem to find a website for NovoCastra labs. Can anyone comment on the Ki-67 antibody from Novocastra specifically for immunohistochemistry. Any help would be greatly appreciated. Thanks, Brain tumor From hodges420 <@t> msn.com Mon Jan 24 06:01:17 2005 From: hodges420 <@t> msn.com (MARY T HODGES) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] sucscribe Message-ID: From dsnider <@t> shrinenet.org Mon Jan 24 06:23:41 2005 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] processing and staining filter membranes Message-ID: Hi Everyone, Does anyone out there have some insight on processing and staining filter membranes from cell cultures? So far I have discovered they must be hand processed and cut thicker than routine tissue. I have come up with a protocol that is working, and the membranes and cells are staying together and adhereing to the slide. So far I have not found a written protocol for this and was curious if anyone in histoland has experience with this and if there are some tips that would make it better. Thanks in advance... Deanna Snider HT Shriners Hospital for Children Research Dept. Cincinnati, OH CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From suda <@t> med.niigata-u.ac.jp Mon Jan 24 09:33:55 2005 From: suda <@t> med.niigata-u.ac.jp (Takeshi Suda) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] anti-luciferase antibody Message-ID: <20050124103355.171CDC98.suda@med.niigata-u.ac.jp> Hi everybody I am looking for a nice anti-luciferase antibody for immunohistochemistry especially after LacZ staining in mouse liver. Thanks a lot for your help! Best regards =========================== Pharmaceutical Sciences University of Pittsburgh Takeshi Suda suda@med.niigta-u.ac.jp =========================== From ddeibler <@t> centexpathlab.com Mon Jan 24 10:36:09 2005 From: ddeibler <@t> centexpathlab.com (David Deibler) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] HT Position Waco,Texas Message-ID: <001201c50232$cfde4d40$2f01010a@tamtron.ctpl.dns> From lyonm <@t> upstate.edu Mon Jan 24 10:56:13 2005 From: lyonm <@t> upstate.edu (Michael J. Lyon) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] developmental myosin antibody Message-ID: I had posted something last week but didn't get any responses. It was likely my subject line. I am looking for a good antibody for developmental myosin that will work in formalin fixed human tissues. Most of the antibodies that I have looked at do not work in fixed tissue. This is a problem since it has become very difficult to get anything that is unfixed in a timely manner. Thanks Michael J. Lyon, Ph.D. Otolaryngology Research Labs SUNY Upstate Medical University 750 East Adams Street Syracuse, NY 13210 Voice 315-464-7253 Fax 315-464-5572 From yej2 <@t> leicester.ac.uk Mon Jan 24 11:12:22 2005 From: yej2 <@t> leicester.ac.uk (El Jai, Y.) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] RE: Rudeness Message-ID: <1F2CE8D4B0195E488213E8B8CCF71486034C9745@Saffron.cfs.le.ac.uk> I am one of the people that had no experience in histology at all until the past few months. Besides barely anyone in my lab or the department is doing any. So when I found the Histonet, and realised I could get advice, it was a great opportunity for me as it is difficult sometimes to to some research via google, without spending hours and without being sure of having found what we need. Having discussions with people who have the experience is most times just as useful if not more than reading a protocol. Fortunately the experience I had was really helpful and I even stayed in touch with another histonetter who sent me more advice later on. I think that is the exact aim of this forum: share experiences and give advice (receive in my case), whatever our background is. It is a real shame that some people get rude answers as it is a place to be communicative and construtive. I hope that I will get some advice and help the next time I need any and would be more than happy to share my experiences with someone who might need it. Happy new year to everyone! Yasmine. From wood <@t> dcpah.msu.edu Mon Jan 24 11:54:41 2005 From: wood <@t> dcpah.msu.edu (Thomas Wood) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] I was looking for Ki-67 antibody forimmunohistochemistry from NovoCastra labs Message-ID: NovaCastra web site: www.novacastra.co.uk -Tom Wood >>> "goutham reddy" 01/23/05 06:26PM >>> I can't seem to find a website for NovoCastra labs. Can anyone comment on the Ki-67 antibody from Novocastra specifically for immunohistochemistry. Any help would be greatly appreciated. Thanks, Brain tumor _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carolb <@t> mail.phys.mcw.edu Mon Jan 24 12:40:42 2005 From: carolb <@t> mail.phys.mcw.edu (Carol Bobrowitz) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] TIMP-1 antibody Message-ID: <59DF6640CDC0D7119DF000D0B761393C1A9621@thor.phys.mcw.edu> Has anyone used TIMP-1 on FFPE rat tissue? If you have, may I ask your source of this primary. Also, I was able to find a antibody to TIMP-2 from a company "DBS" in CA. Does anyone know what DBS stands for? Do companies provide antibodies at a discount if they have not been tested on different species except human tissue? Thank you in advance, Carol Ann Bobrowitz Histology Laboratory Department of Physiology - Room 541 Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, Wisconsin 53226 414-456-8179 FAX 414-456-6546 From dsnider <@t> shrinenet.org Mon Jan 24 12:46:20 2005 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] plastic embedding resins Message-ID: Hello again everyone! I am trying to learn GMA procedures. So far I am just reding and geting familiar with the process. I have heard different opinions on different embedding medias, ie; EMS Embed 812, Spurrs, and the kit the lab has, Technovit 7100. I was also told by different people that Technovit 8100 would be better suited for the area I am in. I would like to have peoples comments on the different products out there. The positive and the negative about the different products. I will be embedding very small, thin biopsies of cultured skin substitute. The processor used is an RMC EMP 5160. Any and all advice and comments welcome.. Thanks Deanna Snider HT Shriners Hospital Cincinnati, OH CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From twheelock <@t> mclean.harvard.edu Mon Jan 24 12:46:00 2005 From: twheelock <@t> mclean.harvard.edu (Timothy R. Wheelock) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] CLEANING GLASSWARE Message-ID: <41F54268.8030000@mclean.harvard.edu> Hi Sue: I use Contrad 70, a liquid detergent, which you can find at Fisher Scientific on-line, or the Fisher catalogue. I dilute it down to 10%, and then immerse my coplin jars and flasks in it (in my case, for the Bielschowsky silver stain), either over the working day or overnight. I change it every 4 staining runs, just as a rule of thumb. You can probably get away with a less than 10% dilution, shorter immersion times, and other changing schedules. It is far safer than the concentrated Nitric Acid that I had been using for years. I had an accident with the Nitric Acid a while ago (major spill), which prompted me to find a safer substitute. Someone on Histonet suggested Contrad 70. It has worked just as well as pure Nitric Acid for my silver stains. They say that you can even dump it down the drain after use, but I always put it in waste container anyway. Check with other people on Histonet though, for their opinions about this product. Tim Wheelock Harvard Brain Tissue Resource Center McLean Hospital Belmont, MA Any information, including protected health information (PHI), transmitted in this email is intended only for the person or entity to which it is addressed and may contain information that is privileged, confidential and or exempt from disclosure under applicable Federal or State law. Any review, retransmission, dissemination or other use of or taking of any action in reliance upon, protected health information (PHI) by persons or entities other than the intended recipient is prohibited. If you received this email in error, please contact the sender and delete the material from any computer. From lteves <@t> uhnres.utoronto.ca Mon Jan 24 14:02:47 2005 From: lteves <@t> uhnres.utoronto.ca (Lucy Teves) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] In situ pcr on paraffin rat brain sections Message-ID: <41F55467.34776BE4@uhnres.utoronto.ca> Hi Our lab is interested in doing In Situ PCR on paraffin rat brain sections on slides. We are trying to identify location of an increase or decrease of assorted proteins. We have arranged for a demo on the Eppendorf Mastercyler which comes with a slide plate. Can any one give me a basic protocol that works so I can test this machine. Thanks in advance Lucy From gcallis <@t> montana.edu Mon Jan 24 14:08:35 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] GMA in RE: plastic embedding resins (long) In-Reply-To: References: Message-ID: <6.0.0.22.1.20050124121958.01b42368@gemini.msu.montana.edu> Deanna, You did not say what kind of application you are using (immunostaining, regular staining for light microscopy, Electron microscopy). This often determines the optimal plastic choice before you launch you tissues into a resin. GMA means glycol methacrylate, and is water miscible however cannot be removed from a section after the plastic is polymerized. EMS embed 812, Spurrs, Araldite, EPON are used (in general) for electron microscopy ultrathin sections and plastic is not removed for staining, unless other than toluidine blue in sodium borate, pH 9 - 11 is used, or methylene blue basic fuchsin. Unless you are doing EM, these plastics may not be ideal. Are you planning to do immunohistochemical staining? If so, methyl methacrylate can be use instead, but the plastic must be totally removed before staining, however, IHC is possible aftert the plastic is gone, something that will NOT happen with GMA, it cannot be removed after polymerization. Technovits 7100 and 8100 (EBS) and JB-4 and JB-4 Plus (Polysciences) are glycol methacrylate kits, and tissue can be stained with most water soluble stains i.e H&E, PAS, etc, etc Immunohistochemistry is poor since GMA cannot be removed from the tissue once polymerized. GMA will not allow large immunoglobulins to reach antigens in tissue, cannot penetrate the plastic. When choosing GMA, I would assume either would work well, 8100 or 7100 - I know the 7100 sections like a dream! I am not sure why you say one is suited better than another for your area, unless you mean humidity. There are ways to counteract that problem when working with GMA. Plastic monomers cannot be used on processors and it would take far too much monomer to fill reservoirs, plus it is toxic and expensive. You should work in a hood at all times with these plastics, and keep them off your skin, they are very sensitizing. ts. Hand processing is generally done OR you can use the processor to dehydrate the tissue (2 mm thick, thicker may lead to polymerization problems) up through absolute ethanol and then do plastic monomer infitrations separately, in a hood and refrigerator. Since tissues are so thin and small, hand processing takes very little time with infiltration done overnight in a refrigerator or even a few hours without vacuum at RT. Early polymerization must be avoided, hence cold temps. If you exclude air, the plastic will polymerize BEFORE you want it to and there is no going back to recover tissue when this happens, been there, had it happen! Huge tearing of hair!! Polysciences should have a processing protocol in their technical sheet, maybe on their website or you can contact pmarcum@polysciences.com for information. For Technovits - go to these websites - both are processing/handling protocols, very nice and tidy. EBS sells Technovits http://www.biol.s.u-tokyo.ac.jp/users/hassei/plotocol/plasti_%20embedding_8100.html http://www.ebsstore.com/control/category/~category_id=E1/~pcategory=E Gamble and Bancrofts Theory and Practice of Histological Technique, second edition, 2002 has a whole chapter devoted to using plastics, i.e. GMA. Be sure to go back in Histonet too, there have been many discussion about plastics. Type in resin names as keywords. There have also been many excellent articles published over the years in J of Histotechnology on glycol methacrylate. If you have a medical library handy and they have the journal, it is worth some digging out to learn about this plastic along with staining methods. etc. J of Histotechnology is available on line for at least the issues. There are special embedding molds from these companies, designed for GMA work (Polysciences) and block holders - these are very handy and worth the investment, molds are resuable. You will need extremely sharp knives, we prefer glass knives although tungsten carbide. Some have success with disposable microtome blades. Good luck At 11:46 AM 1/24/2005, you wrote: >I am trying to learn GMA procedures. So far I am just reding and geting >familiar with the process. I have heard different opinions on different >embedding medias, ie; EMS Embed 812, Spurrs, and the kit the lab has, >Technovit 7100. I was also told by different people that Technovit 8100 >would be better suited for the area I am in. I would like to have peoples >comments on the different products out there. The positive and the negative >about the different products. I will be embedding very small, thin biopsies >of cultured skin substitute. The processor used is an RMC EMP 5160. > >Any and all advice and comments welcome.. >Thanks >Deanna Snider HT >Shriners Hospital >Cincinnati, OH 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 zhuxx003 <@t> umn.edu Mon Jan 24 16:05:21 2005 From: zhuxx003 <@t> umn.edu (Danielle Jin) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] bubbles Message-ID: <41F57121.4BA14859@umn.edu> Hello, I am new to histonet. I hope someone out there can help me solve my problem. When I do muscle staining (eg. acid phosphatase), the last step is use glycerol Gelatin to cover the coverslip. Somehow it always trap some bubbles in the tissue section. How can I get rid of the bubbles? Thanks Danielle Jin From jmahoney <@t> alegent.org Mon Jan 24 16:14:42 2005 From: jmahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] bubbles Message-ID: First be sure there are no bubbles in the glycerol/gelatin. Then be sure that the sliide is still wet (with distilled water) when you coverslip. Hope this helps. Jan Omaha >>> Danielle Jin 01/24/2005 4:05:21 PM >>> Hello, I am new to histonet. I hope someone out there can help me solve my problem. When I do muscle staining (eg. acid phosphatase), the last step is use glycerol Gelatin to cover the coverslip. Somehow it always trap some bubbles in the tissue section. How can I get rid of the bubbles? Thanks Danielle Jin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Mon Jan 24 16:17:20 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] ISH control probe In-Reply-To: Message-ID: <000001c50262$7932a560$76d48a80@AMY> Patti I think I can help you, but I need to know what your target is. Typically we normally are going after the mRNA rather than the DNA that has only one to a few copies and is duplexed. So you would be more concerned about the integrity of RNA rather than DNA. If you are looking for RNA then we use a probe that hits 18s rRNA that hits all eucarya. It's an oligo probe that you can get any oligo company to make up with a fluor or hapten attached to it. Its 5'GGGCATCACAGACCTG3' with Tm = 52?C. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patti Loykasek Sent: Friday, January 21, 2005 12:14 PM To: histonet Subject: [Histonet] ISH control probe I have a question for those folks doing ISH. What type of probe are you using to determine that the DNA in the tissue is available for hybridization? Thanks for the help. Patti Loykasek PhenoPath Laboratories Seattle, WA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Mon Jan 24 17:21:25 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] bubbles Message-ID: Danielle, Did you make the glycerol fresh or make it and let it set over night? I do immunofluorescence and use a 1:10 glycerol/pbs solution. Could you use the same? Robyn OHSU >>> Danielle Jin 1/24/2005 2:05:21 PM >>> Hello, I am new to histonet. I hope someone out there can help me solve my problem. When I do muscle staining (eg. acid phosphatase), the last step is use glycerol Gelatin to cover the coverslip. Somehow it always trap some bubbles in the tissue section. How can I get rid of the bubbles? Thanks Danielle Jin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LuckG <@t> empirehealth.org Mon Jan 24 17:23:26 2005 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Job Opening Message-ID: Good afternoon, We have an opening for a histotech, full time, day shift, Monday through Friday. Routine histology duties (embedding, cutting and special stains). ASCP certification preferred but not required. Anyone not familiar with Spokane can get a good overview by visiting the website www.visitspokane.com . Spokane has a low cost of living and a great quality of life. Anyone interested can apply on-line at www.deaconessmedicalcenter.org . If you want any additional information about the job, the organization or anything else feel free to contact me and remember (I know because it took me some time to learn) "that making a living is not the same thing as making a life". Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org From ronkent3 <@t> comcast.net Mon Jan 24 18:11:13 2005 From: ronkent3 <@t> comcast.net (ron) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] bubbles In-Reply-To: References: Message-ID: <41F58EA1.8050806@comcast.net> Robyn Vazquez wrote: Danielle, Did you make the glycerol fresh or make it and let it set over night? I do immunofluorescence and use a 1:10 glycerol/pbs solution. Could you use the same? Robyn OHSU Danielle Jin [1] 1/24/2005 2:05:21 PM >>> Hello, I am new to histonet. I hope someone out there can help me solve my problem. When I do muscle staining (eg. acid phosphatase), the last step is use glycerol Gelatin to cover the coverslip. Somehow it always trap some bubbles in the tissue section. How can I get rid of the bubbles? Thanks Danielle Jin _______________________________________________ Histonet mailing list [2]Histonet@lists.utsouthwestern.edu [3]http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list [4]Histonet@lists.utsouthwestern.edu [5]http://lists.utsouthwestern.edu/mailman/listinfo/histonet References 1. mailto:zhuxx003@umn.edu 2. mailto:Histonet@lists.utsouthwestern.edu 3. http://lists.utsouthwestern.edu/mailman/listinfo/histonet 4. mailto:Histonet@lists.utsouthwestern.edu 5. http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Lynda.King <@t> vision-bio.com Mon Jan 24 18:55:21 2005 From: Lynda.King <@t> vision-bio.com (Lynda King) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] I was looking for Ki-67 antibody forimmunohistochemistry from NovoCastra labs Message-ID: <5E06BFED29594F4C9C5EBE230DE320C608DEA44A@ewok.vsl.com.au> Try this for the Novocastra website www.novocastra.co.uk Thanks Lynda King -----Original Message----- From: goutham reddy [mailto:greddy9734@hotmail.com] Sent: Monday, 24 January 2005 10:26 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] I was looking for Ki-67 antibody forimmunohistochemistry from NovoCastra labs I can't seem to find a website for NovoCastra labs. Can anyone comment on the Ki-67 antibody from Novocastra specifically for immunohistochemistry. Any help would be greatly appreciated. Thanks, Brain tumor _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hodges420 <@t> msn.com Mon Jan 24 20:25:06 2005 From: hodges420 <@t> msn.com (MARY T HODGES) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] studenta and presentation Message-ID: Good evening all. Does anyone have any ideas for presenting a talk to histo students regarding biopsies, cutting and embedding. Any ideas about serial sections. levels and different cutting techniques used by pathologists for GI biopsy. If you have any articles pictures ect. email me please Thanks to you all, Tere Hodges St Mary's Hospital Tucson, Az. From pathrm35 <@t> adelphia.net Mon Jan 24 20:57:41 2005 From: pathrm35 <@t> adelphia.net (Ron Martin) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Histotech position in Palm Beach Gardens,Florida Message-ID: <000601c50289$a49f8520$27233418@Pathrm35> Small but growing dermatology practice in southeast Florida seeking a Florida licensed/eligible technologist. HT and/or HTL required and a college degree a plus but not required. Early shift (6am-2:30pm) and duties include embedding, sectioning and special stains. This is a great opportunity for an independent ambitious person who wants to grow with an expanding company. Ron Martin, B.Sc., HT (ASCP) HTL Histology Supervisor From hodges420 <@t> msn.com Tue Jan 25 06:40:04 2005 From: hodges420 <@t> msn.com (MARY T HODGES) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] email Message-ID: Can you tell me if I am back on the email list. I have several questains out for info. Thanks tere hodges From arvind <@t> nbrc.res.in Tue Jan 25 07:41:06 2005 From: arvind <@t> nbrc.res.in (Arvind Pundir) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Ache staining querry Message-ID: <7DE0291A47C4BC4D9CDEF65D5017CEF0A714@mail.nbrc.res.in> i do Ache staining frequently on human brain sections, but facing problem that there is no staining at the edges of my section can anyone sort what it may be due to thanks in advance for any suggestion or advice Arvind Singh Pundir National Brain Research Centre INDIA From tom_c_nguyen <@t> yahoo.com Tue Jan 25 10:02:21 2005 From: tom_c_nguyen <@t> yahoo.com (Tom C. Nguyen) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] tissue slicer Message-ID: <20050125160221.46980.qmail@web51107.mail.yahoo.com> dear all, i am a postdoctoral research fellow at stanford university working in the cardiovascular lab. one of our projects involve mapping out the fiber orientation of the heart. we cut select areas of the heart into 1cmx1cmx1cm blocks. i was wondering anyone knew of a device that can reliable cut this heart block into 1mm slices... i guess similar to one of the machines at the deli that slices of a big chunk of turkey into this slices. thanks for your thoughts. bests, -tom http://scalpel.stanford.edu http://surgery.stanford.edu From eca9 <@t> georgetown.edu Tue Jan 25 10:10:41 2005 From: eca9 <@t> georgetown.edu (Eva C Anderson) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Fluorescence staining Message-ID: <41F66F81.2030706@georgetown.edu> Hi, We are looking into trying some Fluorescence staining. We want to try a TSA Fluorescence System from PerkinElmer. The IHC protocol does however leave me with some questions. We always dehydrate our samples after staining. The protocol however doesn't mention this. It only says that the samples can be counterstained and mounted after incubation with the Fluorophore Tyramide. Should we dehydrate and is there a particular mounting media we should use to prolong the time that fluorescence can be detected? Any help would be greatly appreciated. Thanks in advance for all your help, Eva Georgetown University Research Assistant From mcauliff <@t> umdnj.edu Tue Jan 25 13:35:55 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] tissue slicer In-Reply-To: <20050125160221.46980.qmail@web51107.mail.yahoo.com> References: <20050125160221.46980.qmail@web51107.mail.yahoo.com> Message-ID: <41F69F9B.4050401@umdnj.edu> Hi Tom: Sort of depends on how big the heart is. A deli slicer seems like a good idea, as long as it gives reproducable sections. For small hearts (mouse, rat) you could get a bunch of razor blades and glue them together with "super glue" including metal spacers of the appropriate dimensions in between the blades. A lot cheaper that a deli slicer. You could also invest in a "Vibratome" a device for cutting sections of either fixed or unfixed tissue. Since I teach Gross Anatomy I would be very interested in the results of your research! Geoff Tom C. Nguyen wrote: >dear all, >i am a postdoctoral research fellow at stanford university working in the cardiovascular lab. one of our projects involve mapping out the fiber orientation of the heart. we cut select areas of the heart into 1cmx1cmx1cm blocks. i was wondering anyone knew of a device that can reliable cut this heart block into 1mm slices... i guess similar to one of the machines at the deli that slices of a big chunk of turkey into this slices. thanks for your thoughts. >bests, -tom > > > > > >http://scalpel.stanford.edu >http://surgery.stanford.edu > > > > > >_______________________________________________ >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 pwebster <@t> hei.org Tue Jan 25 10:44:55 2005 From: pwebster <@t> hei.org (Paul Webster) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] tissue slicer In-Reply-To: <20050125160221.46980.qmail@web51107.mail.yahoo.com> Message-ID: Dear Tom, I have the most ideal machine for you. A company in Northern California has just sent me a machine that does exactly what you are looking for, slice fresh tissue into 1mm slices. I am currently evaluating it and it is pretty good. He tissue is cut in one pass through the machine much as bread is sliced. I can give you contact details for the makers of this machine (the "Equal-Z) if you are interested. As far as I know, this is a new innovation that is not currently on the market. I have no links with the company other than agreeing to be a beta test site. Regards, Paul Webster. Paul Webster, Ph.D. Scientist II and Director Ahmanson Advanced EM and Imaging Center House Ear Institute 2100 West Third Street Los Angeles CA 90057-1922 Phone: 213 273 8026 Fax: 213 13 739 E-mail: pwebster@hei.org On 1/25/05 8:02 AM, "Tom C. Nguyen" wrote: > dear all, > i am a postdoctoral research fellow at stanford university working in the > cardiovascular lab. one of our projects involve mapping out the fiber > orientation of the heart. we cut select areas of the heart into 1cmx1cmx1cm > blocks. i was wondering anyone knew of a device that can reliable cut this > heart block into 1mm slices... i guess similar to one of the machines at the > deli that slices of a big chunk of turkey into this slices. thanks for your > thoughts. > bests, -tom > > > > > > http://scalpel.stanford.edu > http://surgery.stanford.edu > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gcallis <@t> montana.edu Tue Jan 25 11:52:03 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Fluorescence staining In-Reply-To: <41F66F81.2030706@georgetown.edu> References: <41F66F81.2030706@georgetown.edu> Message-ID: <6.0.0.22.1.20050125104715.01afd888@gemini.msu.montana.edu> Molecular Probes has mounting media designed to set up hard but mainly prevent photobleaching of fluorophore, in your case FITC? Go to their website and look for Prolong Gold, ready to use. We love it. You mount the sections from PBS, and do not dehydrate. They recommend sealing the edges of coverslip, which is done nicely with toluene or xylene diluted mounting media at consistency of very thin nail polish. We avoid latter due to alcohol in nail polish. It is pricey but worth every penny of cost. At 09:10 AM 1/25/2005, you wrote: >Hi, >We are looking into trying some Fluorescence staining. We want to try a >TSA Fluorescence System from PerkinElmer. The IHC protocol does however >leave me with some questions. >We always dehydrate our samples after staining. The protocol however >doesn't mention this. It only says that the samples can be counterstained >and mounted after incubation with the Fluorophore Tyramide. Should we >dehydrate and is there a particular mounting media we should use to >prolong the time that fluorescence can be detected? Any help would be >greatly appreciated. >Thanks in advance for all your help, >Eva >Georgetown University >Research Assistant 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 deb.vaneyck <@t> phci.org Tue Jan 25 12:59:04 2005 From: deb.vaneyck <@t> phci.org (Van Eyck, Deb) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] cd15 and morphology on bone marrows and lymph nodes Message-ID: Hi all, 1.I would really appreciate some replies to a couple of questions. One is CD15; I thought I had a pretty good antibody going, but it is still problematic in some cases. I would like to know what others are using consistently with heme path approval. The same goes for kappa and lambda and clonality. 2. The other issue is handling bone marrows and lymph nodes---we have recently gotten rid of B-5 and we use NBF as our main fixative. I would like some replies in regards to what is the best fixation/decal situation for morphology purposes (does anyone feel that they have reached the b-5 nirvana state for good morphology) and still maintain immuno staining???? Times and protocols and products would be great. Thanks----Deb Van Eyck-Anatomic Path Waukesha Memorial Hospital 725 American Ave. Waukesha,WI 53188 262-928-2112 fax:262-928-4849 This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. From jbirkner <@t> colabserv.com Tue Jan 25 13:17:01 2005 From: jbirkner <@t> colabserv.com (Jeff Birkner) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] reagent pricing Message-ID: What is everyone paying for reagent alcohol, xylene and formalin? We are paying the following per gallon, not including shipping: Reagent alcohol $15.00 Xylene $28.00 10% NB Formalin $5.45 Jeff Birkner, CT(ASCP) Pathology Section Manager Collaborative Laboratory Services, LLC 1005 Pennsylvania Ave. Ottumwa, IA 52501 From jbirkner <@t> colabserv.com Tue Jan 25 13:56:47 2005 From: jbirkner <@t> colabserv.com (Jeff Birkner) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] RE: reagent pricing Message-ID: I will put the responses in an excel spreadsheet and share them with anyone that wants them. _____ From: Jeff Birkner Sent: Tuesday, January 25, 2005 1:17 PM To: 'histonet@lists.utsouthwestern.edu' Subject: reagent pricing What is everyone paying for reagent alcohol, xylene and formalin? We are paying the following per gallon, not including shipping: Reagent alcohol $15.00 Xylene $28.00 10% NB Formalin $5.45 Jeff Birkner, CT(ASCP) Pathology Section Manager Collaborative Laboratory Services, LLC 1005 Pennsylvania Ave. Ottumwa, IA 52501 From juan.gutierrez <@t> christushealth.org Tue Jan 25 14:14:17 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] reagent pricing Message-ID: Reagent alcohol 8.73/gal Xylene 7.85/gal Formalin 2.92/gal Say Hi to Radar for me. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jeff Birkner Sent: Tuesday, January 25, 2005 1:17 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] reagent pricing What is everyone paying for reagent alcohol, xylene and formalin? We are paying the following per gallon, not including shipping: Reagent alcohol $15.00 Xylene $28.00 10% NB Formalin $5.45 Jeff Birkner, CT(ASCP) Pathology Section Manager Collaborative Laboratory Services, LLC 1005 Pennsylvania Ave. Ottumwa, IA 52501 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Tue Jan 25 14:49:42 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Ache staining querry References: <7DE0291A47C4BC4D9CDEF65D5017CEF0A714@mail.nbrc.res.in> Message-ID: <41F6B0E6.5B99D9E2@uwo.ca> Assuming "Ache staining" is acetylcholinesterase activity histochemistry, not a painful bruise: The surface of the specimen will have been fixed more than the inside. AChE activity resists short fixation in formaldehyde (such as overnight). Perhaps the outside of the block has had more than is good for the enzyme. In small specimens of animal tissues, both the choline esterases (and other esterases) are well preserved by overnight fixation in formaldehyde. Human brains need much longer because of the large size, and this can be expected to wreck enzyme activities. You might still be able to stain the AChE protein with an immunohistochemical method. -- ------------------------------- 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 _______________________________ Arvind Pundir wrote: > > i do Ache staining frequently on human brain sections, but facing problem that there is no staining at the edges of my section can anyone sort what it may be due to > > > > thanks in advance for any suggestion or advice > > > > Arvind Singh Pundir > > National Brain Research Centre > > INDIA > > ------------------------------------------------------------ > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Patty.Lott <@t> ORTHO.UAB.EDU Tue Jan 25 16:30:45 2005 From: Patty.Lott <@t> ORTHO.UAB.EDU (Patty Lott) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Rank-L Message-ID: <85F6C7A1330E794DB8540AFD001CC77E05328DA6@rosco.ortho.uab.edu> Does anyone know of an antibody for RANK-L to be used on FFPE mouse bones? Patty Lott, Laboratory Supervisor Orthopaedic Research Laboratory Center for Metabolic Bone Disease Laboratory University of Alabama at Birmingham LHRB B37 0007 1919 7th Ave. South Birmingham, AL 35294 (205) 934-2007 From lhartman18 <@t> adelphia.net Tue Jan 25 19:38:45 2005 From: lhartman18 <@t> adelphia.net (Linda Hartman) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Job opportunity central PA Message-ID: <002501c50347$c7335a80$1d04a245@linda47p53fn9e> Mount Nittany Medical Center in State College, PA has an opening for a full time histology technician. The applicant will be responsible for routine histology, frozen sections, special staining and IHC. Mount Nittany Medical Center is a growing regional medical center located mid-way between Philadelphia and Pittsburgh. State College is the home of the main campus of Penn State University. Please check out our website at www.mountnittany.org Linda Hartman HT(ASCP) Histology section head Mount Nittany Medical Center State College, PA 16803 Phone: 814-234-6117 email: lhartman@mountnittany.org From amosbrooks <@t> earthlink.net Tue Jan 25 20:07:30 2005 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Freezing Biopsies Message-ID: <6.2.0.14.0.20050125210242.01da7198@pop.earthlink.net> Hi, I'm snap freezing kidney biopsies in OCT using isopentane that is chilled with liquid nitrogen. I'm having a problem where the area that the biopsy is gets raised up above the surface of the block. I'm not sure what is going wrong and more importantly what to do about it. Has anyone had similar problems and how did you solve it? Thanks, Amos Brooks From kappeler <@t> patho.unibe.ch Wed Jan 26 00:52:43 2005 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] cd15 and morphology on bone marrows and lymph nodes References: Message-ID: <002401c50373$a3254be0$27955c82@patho.unibe.ch> Hi Deb We use mo-a-CD15, clone MMA, Becton-Dickinson 347420. Working conc. is 2.5 ug Ig/ml, pretreatment is HIER (citrate) in pressure cooker. The 'secret' with this clone (and most other commercially available CD15 mAbs, such as C3D-1, BY87, 80H5, and more) is, that it is of Ig class IgM (and not IgG1, IgG2a, as most monoclonals that you commonly use). While most secondary antibodies show some cross-reactivity with mAbs of Ig class IgM, you have to use a mouse-IgM specific secondary antibody to get optimal results (e.g. DakoCytomation E 0465, rb-a-mo IgM/Biotin). This applies also to a couple of other mAbs: they will work with 'regular' secondary antibodies, however, they will work better with an IgM-specific secondary antibody. Therefore it is always useful to check the SpecSheet for the Ig-class of a given mAb. If you are using a visualization system that comes from the supplier of your staining machine, you may have a problem, depending on the stainer. Some of them are obviously very reluctant to let you use secondary reagents of your choice... Then it is good to know, that immunos can be done manually ... Hope this helps. Good luck. Andi Kappeler Insitute of Pathology, University of Bern, Switzerland ----- Original Message ----- From: "Van Eyck, Deb" To: Sent: Tuesday, January 25, 2005 7:59 PM Subject: [Histonet] cd15 and morphology on bone marrows and lymph nodes > > Hi all, > 1.I would really appreciate some replies to a couple of questions. One is > CD15; I thought I had a pretty good antibody going, but it is still > problematic in some cases. I would like to know what others are using > consistently with heme path approval. The same goes for kappa and lambda > and clonality. > > > 2. The other issue is handling bone marrows and lymph nodes---we have > recently gotten rid of B-5 and we use NBF as our main fixative. I would > like some replies in regards to what is the best fixation/decal situation > for morphology purposes (does anyone feel that they have reached the b-5 > nirvana state for good morphology) and still maintain immuno staining???? > Times and protocols and products would be great. > > > Thanks----Deb Van Eyck-Anatomic Path > Waukesha Memorial Hospital > 725 American Ave. > Waukesha,WI 53188 > 262-928-2112 > fax:262-928-4849 > > > > This information is confidential and intended solely for the use of the > individual or entity to whom it is addressed. > If you have received this email in error please notify the sender or our > Customer Support Center at (262) 928-2777. > > We have scanned this email and its attachments for malicious content. > However, the recipient should check this email and any attachments for the > presence of viruses. > ProHealth Care accepts no liability for any damage caused by any virus > transmitted by this email. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ctsblack <@t> capeheart.uct.ac.za Wed Jan 26 01:21:12 2005 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Eluting Solution - Glycin-HCl Message-ID: Hi There Can anybody give me a method for making up Glycin-HCl(0.1M, pH 2.2) to be used an a blocking buffer in double immuno staining??? 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 c.m.vanderloos <@t> amc.uva.nl Wed Jan 26 01:52:21 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] RE: Fluorescence staining Message-ID: <12c276c12c2991.12c299112c276c@amc.uva.nl> Dear Eva, To my knowledge most of the traditional fluorochromes like FITC, rhodamine, doesn't stand dehydration at all. So in your case it fully depends on what type of fluorochrome you end up with after the TSA procedure. If you end up with FITC, the use of VectaShield ([1]www.vectorlabs.com) for mounting is highly recommended for the prevention of photobleaching. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: [2]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From Eva C Anderson Date Tue, 25 Jan 2005 11:10:41 -0500 To histonet@lists.utsouthwestern.edu Subject [Histonet] Fluorescence staining Hi, We are looking into trying some Fluorescence staining. We want to try a TSA Fluorescence System from PerkinElmer. The IHC protocol does however leave me with some questions. We always dehydrate our samples after staining. The protocol however doesn't mention this. It only says that the samples can be counterstained and mounted after incubation with the Fluorophore Tyramide. Should we dehydrate and is there a particular mounting media we should use to prolong the time that fluorescence can be detected? Any help would be greatly appreciated. Thanks in advance for all your help, Eva Georgetown University Research Assistant References 1. http://www.vectorlabs.com/ 2. mailto:c.m.vanderloos@amc.uva.nl From carl.hobbs <@t> kcl.ac.uk Wed Jan 26 02:09:43 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] testing connection Message-ID: <000301c5037e$646dfea0$e8345c9f@Carlos> My emails keep getting bounced From wayneholland1959 <@t> msn.com Wed Jan 26 06:10:50 2005 From: wayneholland1959 <@t> msn.com (Wayne Holland) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] subscribe to the histonet please, thanks Message-ID: _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From Tbarnhart <@t> primecare.org Wed Jan 26 07:29:02 2005 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Hairy Cell Leukemia Spleen Tissue Message-ID: <1779904B5E82D511914C00D0B793339205BFD8B1@exchangent> I am in need of the above control tissue. We have not had a case here that we can find and would like to use it as a control for our TRAcP antibody. Willing to pay, hopefully not too much. Or, trade for other tissue :) Thanks.... Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND tbarnhart@primecare.org 701-530-6730 Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. From dobbin <@t> upei.ca Wed Jan 26 06:48:59 2005 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Attn: Roxanne Soto & Emily Smith Message-ID: <41F7678B.22027.1B921F@localhost> Roxanne and Emily, I am getting planning underway again for the Histonet Outpost. I sent out an e-mail to our `ad hoc' committee members yesterday and your e-mails bounced. Can you confirm that your e-mail addresses (below) are correct and if not, provide the proper one so I can forward you the e-mail again as well as all the replies I received to it. Thanks. Greg ESmith@CuraGen.com GREYTRUNK@aol.com ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Research Technologist, Pathology Lab, Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From colins <@t> thor.uk.com Wed Jan 26 09:07:05 2005 From: colins <@t> thor.uk.com (Colin Smith) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Manual fixation/processing of reconstructed skin cultures grown on polycarbonate filters Message-ID: <3E985CD1D7CA89499D1902BC3202AF3CC1A75E@uk-res-srv-03.thorspecialities.com> Can anyone help? I am currently setting up a basic histology system for analysing reconstructed tissues grown in culture on polycarbonate filters, similar to Mattek's Epiderm. We currently have begged borrowed or stolen, equipment-wise: two water baths (ambient - 100C), a circulating warm air oven, a rocking microtome and cold blocks, along with glass staining jars/racks, forceps etc. At the moment, equipment such as a wax dispenser, purpose built cooling plates etc are out of the question until I can show that we can show that the model we are growing is validated, which will include immunohistochemistry of cytokeratin and cell adhesion markers etc. Thus we are in the catch 22 situation of having to prove something without the equipment needed to do the job, in order to justify buying the equipment!! But thats accountants for you, go figure. Anyway, can anyone give any useful pointers/protocols for fixing/processing/embedding/sectioning such tissues using such primative equipment Any help, no matter how trivial will be greatfully received. Many Thanks in Advance, Colin Smith BSc CBiol MIBiol Eurotox Registered Toxicologist In Vitro Toxicologist Thor Specialities UK Ltd Wincham Avenue, Northwich CW9 6GB Tel: +44 1606 818873 Fax: +44 1606 818801 colins@thor.uk.com The information contained in this e-mail may be confidential and may also be legally privileged. It is intended solely for the addressee(s). If you are not the intended recipient then please delete forthwith. Views expressed in this email are views of the individual and are not necessarily the views of this company. ************************************************************************************************* e-mail scanned for Viruses by Thor Specialities (UK) Limited using Gfi Mail Security ************************************************************************************************* From kspencer <@t> utmem.edu Wed Jan 26 09:39:59 2005 From: kspencer <@t> utmem.edu (Kathleen Spencer) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Freezing Biopsies In-Reply-To: <6.2.0.14.0.20050125210242.01da7198@pop.earthlink.net> References: <6.2.0.14.0.20050125210242.01da7198@pop.earthlink.net> Message-ID: <89982A33-6FB0-11D9-8BD6-000393967904@utmem.edu> Amos, The way we did this was to place the biopsy in a plastic mold, add OCT, grab the huge forceps and plunge it into liquid nitrogen and count to 5 or 10, I can't remember, but it always worked fine. We did not use isopentane. The surface of the block was always nice and smooth. It had to be! The biopsies were so tiny. What you can do is get a fresh mold, put a little OCT in the bottom, put the frozen block in and spray it a little with cryo spray. You should now have a smooth surface for sectioning. Good Luck! Kathleen Spencer HT (ASCP) Lab Manager/LCM Supervisor UTHSC On Jan 25, 2005, at 8:07 PM, Amos Brooks wrote: > Hi, > I'm snap freezing kidney biopsies in OCT using isopentane that is > chilled with liquid nitrogen. I'm having a problem where the area that > the biopsy is gets raised up above the surface of the block. I'm not > sure what is going wrong and more importantly what to do about it. Has > anyone had similar problems and how did you solve it? > > Thanks, > Amos Brooks > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kappeler <@t> patho.unibe.ch Wed Jan 26 10:18:31 2005 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] cd15 and morphology on bone marrows and lymph nodes References: Message-ID: <006301c503c2$ad285ed0$27955c82@patho.unibe.ch> We use the rb-a-mo IgM/Biotin in stead of our usual anti-mouse Ig secondary antibody (which is gt-a-mo Ig/Biotin). As we still stain manually (yes that exists...!) this is no problem. Adding the rb-a-mo IgM to your usual secondary is theoretically possible, however if your secondary is a 'multi link' reagent (anti-mouse and anti-rabbit cocktail) then you should not add a rb-a-mo IgM/Biotin as it will be bound by the anti-rb component of your 'multi link' reagent. The system may still work, but I would prefer to know what I'm doing rather than to 'let something happen' inside a tube where I don't have control. If you want to add an anti-mouse IgM component to your 'multi link', make sure it does not intefere with any of the components that is already in there (e.g. use a gt-a-mo IgM/Biotin). Our protocol for CD15 looks as follows (in brief): 1. Deparaffinization --> TBS 2. Pretreatment, citrate buffer, pressure cooker, 7 min at 121 C, 1 bar; cool down, wash 3. Incubate with mo-a-CD15, clone MMA, 2.5 ug Ig/ml 4. Wash 5. Incubate with rb-a-mo IgM (DakoCytomation E 0465, 1:500) 6. Wash 7. Incubate with enzyme system (ABC/HRP, LSAB+/HRP, whatever...) 8. Wash 9. Develop with 3,3'-diaminobenzidin /H2O2 10. Rinse, counterstain, mount Best regards, Andi ----- Original Message ----- From: "Van Eyck, Deb" To: "'Andi Kappeler'" Sent: Wednesday, January 26, 2005 4:20 PM Subject: RE: [Histonet] cd15 and morphology on bone marrows and lymph nodes > thanks very much-------for the secondary system do you just use this with > another Dako product for example---are you using lsab or lsab +. I know > some people use this system and just add the IgM to the secondary--what > specifically do you do. this has been a real help Andi! > > > -----Original Message----- > > From: Andi Kappeler [SMTP:kappeler@patho.unibe.ch] > > Sent: Wednesday, January 26, 2005 12:53 AM > > To: Histonet; Van Eyck, Deb > > Subject: Re: [Histonet] cd15 and morphology on bone marrows and lymph > > nodes > > > > Hi Deb > > > > We use mo-a-CD15, clone MMA, Becton-Dickinson 347420. Working conc. is 2.5 > > ug Ig/ml, pretreatment is HIER (citrate) in pressure cooker. The 'secret' > > with this clone (and most other commercially available CD15 mAbs, such as > > C3D-1, BY87, 80H5, and more) is, that it is of Ig class IgM (and not > > IgG1, > > IgG2a, as most monoclonals that you commonly use). While most secondary > > antibodies show some cross-reactivity with mAbs of Ig class IgM, you > > have > > to use a mouse-IgM specific secondary antibody to get optimal results > > (e.g. > > DakoCytomation E 0465, rb-a-mo IgM/Biotin). This applies also to a couple > > of > > other mAbs: they will work with 'regular' secondary antibodies, however, > > they will work better with an IgM-specific secondary antibody. Therefore > > it > > is always useful to check the SpecSheet for the Ig-class of a given mAb. > > If you are using a visualization system that comes from the supplier of > > your > > staining machine, you may have a problem, depending on the stainer. Some > > of > > them are obviously very reluctant to let you use secondary reagents of > > your > > choice... Then it is good to know, that immunos can be done manually ... > > Hope this helps. Good luck. > > > > Andi Kappeler > > Insitute of Pathology, University of Bern, Switzerland > > > > > > > > ----- Original Message ----- > > From: "Van Eyck, Deb" > > To: > > Sent: Tuesday, January 25, 2005 7:59 PM > > Subject: [Histonet] cd15 and morphology on bone marrows and lymph nodes > > > > > > > > > > Hi all, > > > 1.I would really appreciate some replies to a couple of questions. One > > is > > > CD15; I thought I had a pretty good antibody going, but it is still > > > problematic in some cases. I would like to know what others are using > > > consistently with heme path approval. The same goes for kappa and > > lambda > > > and clonality. > > > > > > > > > 2. The other issue is handling bone marrows and lymph nodes---we have > > > recently gotten rid of B-5 and we use NBF as our main fixative. I would > > > like some replies in regards to what is the best fixation/decal > > situation > > > for morphology purposes (does anyone feel that they have reached the b-5 > > > nirvana state for good morphology) and still maintain immuno > > staining???? > > > Times and protocols and products would be great. > > > > > > > > > Thanks----Deb Van Eyck-Anatomic Path > > > Waukesha Memorial Hospital > > > 725 American Ave. > > > Waukesha,WI 53188 > > > 262-928-2112 > > > fax:262-928-4849 > > > > > > > > > > > > This information is confidential and intended solely for the use of the > > > individual or entity to whom it is addressed. > > > If you have received this email in error please notify the sender or our > > > Customer Support Center at (262) 928-2777. > > > > > > We have scanned this email and its attachments for malicious content. > > > However, the recipient should check this email and any attachments for > > the > > > presence of viruses. > > > ProHealth Care accepts no liability for any damage caused by any virus > > > transmitted by this email. > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > This information is confidential and intended solely for the use of the > individual or entity to whom it is addressed. > If you have received this email in error please notify the sender or our > Customer Support Center at (262) 928-2777. > > We have scanned this email and its attachments for malicious content. > However, the recipient should check this email and any attachments for the > presence of viruses. > ProHealth Care accepts no liability for any damage caused by any virus > transmitted by this email. > > From KarBieber <@t> aol.com Wed Jan 26 10:23:56 2005 From: KarBieber <@t> aol.com (KarBieber@aol.com) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] RMSF Message-ID: <145.3e0c9baa.2f291e1c@aol.com> Does anyone have a good source for an antibody for RMSF? Thanks, Karen From gcallis <@t> montana.edu Wed Jan 26 10:25:16 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Freezing Biopsies In-Reply-To: <6.2.0.14.0.20050125210242.01da7198@pop.earthlink.net> References: <6.2.0.14.0.20050125210242.01da7198@pop.earthlink.net> Message-ID: <6.0.0.22.1.20050126085604.01af5e68@gemini.msu.montana.edu> Amos, We experienced this too. We solved by using Tissue Tek plastic embedding molds 10 x 10 mm size. Cut excess edges for a better heat sink around embedding well except for a small tab to hold onto. Carefully pick a mold that is not deformed already - look for the flat ones, not rounded! Tissue Tek has newer round molds that do not require trimming, very nice, tidy and FLAT! Unfortunately, other brands of plastic molds we tried had thicker plastic compared to Tissue Tek molds and blocks twisted out at freezing. Coat bottom of mold with a THIN film of OCT. Biopsy is laid out so it is as flat as possible in OCT film on bottom. Carefully, slowly - add more OCT down corner of mold or away from top of biopsy to keep it from floating up. Overfilling with OCT results in cracked blocks; fill to top of well. We tilt the beaker with cold isopentane , and introduce mold so bottom makes contact with isopentant first. When OCT starts to turn white ON BOTTOM, immerse fully -about 10 sec or so. Bottom freezing first anchors biopsy and keeps it flat, no lift up syndrome. If frozen block face has any deformity to it caused by a funky mold - just smear a thin layer of OCT over block face to fill in these depressions, etc. to have a flatter block face before trimming. Thin layer is important as we tried to avoid any excess warming of block once it is frozen solid. Have fun!! At 07:07 PM 1/25/2005, you wrote: >Hi, > I'm snap freezing kidney biopsies in OCT using isopentane that is > chilled with liquid nitrogen. I'm having a problem where the area that > the biopsy is gets raised up above the surface of the block. I'm not sure > what is going wrong and more importantly what to do about it. Has anyone > had similar problems and how did you solve it? > >Thanks, >Amos Brooks > > > >_______________________________________________ >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 Wed Jan 26 10:33:09 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] RE: Fluorescence staining In-Reply-To: <12c276c12c2991.12c299112c276c@amc.uva.nl> References: <12c276c12c2991.12c299112c276c@amc.uva.nl> Message-ID: <6.0.0.22.1.20050126092847.01b583c0@gemini.msu.montana.edu> Chris and Eva, Vectashield also come in a Hard Set, so you get permanent coverslipping (no coverglass sliding around!) as does the Molecular Probes Prolong Gold ready to use. If you want a counterfluorescence marker for nuclei, and have the appropriate DAPI filter on microscope, you can buy these with DAPI which "counterstains via fluorescence" the DNA in cells. At 12:52 AM 1/26/2005, you wrote: > Dear Eva, > > To my knowledge most of the traditional fluorochromes like FITC, > rhodamine, doesn't stand dehydration at all. So in your case it fully > depends on what type of fluorochrome you end up with after the TSA > procedure. > > If you end up with FITC, the use of VectaShield > ([1]www.vectorlabs.com) for mounting is highly recommended for > the prevention of photobleaching. > > Chris van der Loos, PhD > Dept. of Pathology > Academical Medical Center M2-230 > Meibergdreef 9 > NL-1105 AZ Amsterdam > The Netherlands > > phone: +31 20 5665631 > fax: +31 20 6960389 > e-mail: [2]c.m.vanderloos@amc.uva.nl 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 ajennings <@t> unmc.edu Wed Jan 26 10:33:22 2005 From: ajennings <@t> unmc.edu (ajennings@unmc.edu) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Freezing Biopsies In-Reply-To: <89982A33-6FB0-11D9-8BD6-000393967904@utmem.edu> Message-ID: Bernice from Instrumedics I am sure can give you better insight into what use to be called "Gentle Jane" freezing. Helps with heat extraction and overall consistent freezing. It may help. I use to have the same problem with whole mouse embryos, because they were freezing from the outside in and by the time the liver froze/expanded it would pop out of the block. I know biopsies are by far smaller but it could be the same general theory with the OCT freezing from the outside around the sample and the sample having no place else to expand. It actually helped me to freeze in the lN2 vapor instead of dropping the sample into isopentane. The time difference was negligible and I didn't get that pop out peak that I believe you are referring to. The freezing occurred from the bottom of the boat to the top instead from all sides at once. It might not be appropriate for all but it has worked in my lab for many many years. just sharing a thought On Jan 25, 2005, at 8:07 PM, Amos Brooks wrote: > Hi, > I'm snap freezing kidney biopsies in OCT using isopentane that is > chilled with liquid nitrogen. I'm having a problem where the area that > the biopsy is gets raised up above the surface of the block. I'm not > sure what is going wrong and more importantly what to do about it. Has > anyone had similar problems and how did you solve it? > > Thanks, > Amos Brooks From liz <@t> premierlab.com Wed Jan 26 10:36:26 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Manual fixation/processing of reconstructed skincultures grown on polycarbonate filters In-Reply-To: <3E985CD1D7CA89499D1902BC3202AF3CC1A75E@uk-res-srv-03.thorspecialities.com> Message-ID: <000001c503c5$2e345770$76d48a80@AMY> Colin We have worked quite a bit with the epioral and epigingival cultures from Mattek. We have stained with H&E, Ki-67 and Cleaved caspase 3. We processed these specimens on a tissue processor, but you will be able to do this by hand. I'm not sure of the size of your culture wells, but we used an 8mm biopsy punch to remove the constructs from the wells, processed the construct whole between two biopsy pads and bisected with a razor blade prior to embedding on end. As far as fixation 10% NBF will work. The wells were labeled on the side and the entire well was placed in a 50 ml conical tube with fixative. After adequate fixation the specimens were grossed, processed and embedded into paraffin. We processed during the day (not overnight) at 15 minutes per station. We sectioned at 4 microns and stained with both H&E and various Immunohistochemical stains, I can send you images if you are interested. If you need more info, give me a call or e-mail me back. Also we are writing an article that will be in the spring edition of Histologic that basically covers the methods (both processing and analysis) we used to assess proliferation in these in vitro tissue models. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Colin Smith Sent: Wednesday, January 26, 2005 8:07 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Manual fixation/processing of reconstructed skincultures grown on polycarbonate filters Can anyone help? I am currently setting up a basic histology system for analysing reconstructed tissues grown in culture on polycarbonate filters, similar to Mattek's Epiderm. We currently have begged borrowed or stolen, equipment-wise: two water baths (ambient - 100C), a circulating warm air oven, a rocking microtome and cold blocks, along with glass staining jars/racks, forceps etc. At the moment, equipment such as a wax dispenser, purpose built cooling plates etc are out of the question until I can show that we can show that the model we are growing is validated, which will include immunohistochemistry of cytokeratin and cell adhesion markers etc. Thus we are in the catch 22 situation of having to prove something without the equipment needed to do the job, in order to justify buying the equipment!! But thats accountants for you, go figure. Anyway, can anyone give any useful pointers/protocols for fixing/processing/embedding/sectioning such tissues using such primative equipment Any help, no matter how trivial will be greatfully received. Many Thanks in Advance, Colin Smith BSc CBiol MIBiol Eurotox Registered Toxicologist In Vitro Toxicologist Thor Specialities UK Ltd Wincham Avenue, Northwich CW9 6GB Tel: +44 1606 818873 Fax: +44 1606 818801 colins@thor.uk.com The information contained in this e-mail may be confidential and may also be legally privileged. It is intended solely for the addressee(s). If you are not the intended recipient then please delete forthwith. Views expressed in this email are views of the individual and are not necessarily the views of this company. ************************************************************************ ************************* e-mail scanned for Viruses by Thor Specialities (UK) Limited using Gfi Mail Security ************************************************************************ ************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From migueljarrin <@t> yahoo.com Wed Jan 26 11:48:38 2005 From: migueljarrin <@t> yahoo.com (Jarrin Miguel) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] mouse eye lens Message-ID: <20050126174838.64967.qmail@web41901.mail.yahoo.com> Dear all, Let me introduce myself. My name is Miguel and I am PhD student. I am considering to carry out immunochemistry and in situ hybridization studies in parafin sections of eye lens in adult mouse. I would like receive futher information about the best fixative that use. I have certain experience with Paraformaldehide, but I have been advised about Davison's fixative and Penfix. I never have heared about Penfix before. Somebody has protocols or receipt for the penfix? Thanking in advance for your help Miguel --------------------------------- Do you Yahoo!? Take Yahoo! Mail with you! Get it on your mobile phone. From jkiernan <@t> uwo.ca Wed Jan 26 12:26:10 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Eluting Solution - Glycin-HCl References: Message-ID: <41F7E0C2.65F46AF7@uwo.ca> Glycine-HCl buffer, pH 2.2 Glycine (0.2M=15.01 g/L): 50 ml HCl (0.2M): 44 ml Water to make 200 ml. >From Lillie & Fullmer 1976. Histopathologic Technic ... 4th edn p.881, where the original references (Sorensen 1909 - 2 papers in a German journal) are given. Approximately 0.2M HCl can be made by 50-60-fold dilution of the concentrated (10-12M) acid. pH 2.2 is lowest item in the glycine-HCl buffer table, so adjustment will probably be needed after making the mixture. -- ------------------------------- 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 _______________________________ Melanie Black wrote: > > Hi There > > Can anybody give me a method for making up Glycin-HCl(0.1M, pH 2.2) > to be used an a blocking buffer in double immuno staining??? > > 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 From bosborn <@t> hsc.usf.edu Wed Jan 26 12:57:08 2005 From: bosborn <@t> hsc.usf.edu (Barbara Osborn) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Re: I was looking for Ki-67 antibody for immunohistochemistry from NovoCastra labs References: <200501241814.j0OIEGS00324@hsc.usf.edu> Message-ID: <00fe01c503d8$d6247560$9d46f783@hscnet.hsc.usf.edu> I use Ki-67 (MM1) mouse monoclonal from NC with great results on FFPE . Citrate boiling for 20 min./ cool down 20 min. used at 1:100 for 1 hour RT. I use M.O.M peroxidase kit from Vector labs on mouse tissue, and Elite ABC peroxidase kit from Vector on other species. > Message: 2 > Date: Sun, 23 Jan 2005 23:26:19 +0000 > From: "goutham reddy" > Subject: [Histonet] I was looking for Ki-67 antibody for > immunohistochemistry from NovoCastra labs > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; format=flowed > > I can't seem to find a website for NovoCastra labs. Can anyone comment on > the Ki-67 antibody from Novocastra specifically for immunohistochemistry. > Any help would be greatly appreciated. > > Thanks, > Brain tumor > > > > > > From m-degutes <@t> northwestern.edu Wed Jan 26 16:22:43 2005 From: m-degutes <@t> northwestern.edu (Mathew DeGutes) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Luxol Fast Blue tissue damage Message-ID: <200501262222.j0QMMtVi028958@casbah.it.northwestern.edu> Hey all any help is appreciated. I am doing some myelin staining with luxol fast blue and have found far too often that I get regular large "holes" in my tissue after staining. My basic protocol is this: -Cryosections on slides rinse in ddH20 then PBS then 70% ethanol -Place in .1% Luxol Fast Blue in 95% Ethanol and .05% Acetic Acid over night at 55C -Remove from LFB and wash in ddH20 then PBS -Differentiate in .05% Lithium Carbonate (usually about 30 sec) -Wash in 70% ethanol and then counterstain or dehydrate and coverslip The actual staining looks pretty good, but I just get these Round white holes as if something bubbled up there and destroyed the tissue. Nothing is boiling so I am curious if anyone else has run into this problem and what might ameliorate it. Thanks again. From komuves <@t> corgentech.com Wed Jan 26 19:09:26 2005 From: komuves <@t> corgentech.com (Laszlo Komuves) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] RE: how to search for antibodies (a user's advice) Message-ID: Dear Histonetters, There have been a lot of postings about antibody sources recently. I would highly recommend to search the following resources: ABCAM ( http://www.abcam.com ) : The search is free but you have to sign up. It is an excellent resource albeit it is biased towards their own antibodies (BTW, I have used many of them, they are excellent) or partner (?) companies (such as Novus). Linscott's directory ( http://www.linscottsdirectory.com ) : There is a yearly membership fee. The database is enormous, well-updated, with links to manufacturers and/or vendors. I used to use a third one (something like MSDS) but somehow lost connection to it (if any one of you can direct me to this site, greatly appreciated. In addition, there are several biomedical resource sites ( I use Biosource, http://www.biocompare.com ), where you can find antibodies. Also there are not that many immunoreagent suppliers. My list includes about 30 companies and I probably buy a wider range of antibodies than most labs because of the diverse interest/need at our company. It is not that difficult to search those major suppliers. For a recent inquire, I run a Linscott's search and found several anti-mouse RANK-L antibodies made by R&D Systems ( http://www.rndsystems ). In fact, knowing their product line and their emphasis on cytokine/chemokine/signalling Abs, I should have searched their site first. If you search these databases, and build your on database (just add the links of your major suppliers to your favorites folder in your browser), you will find most of the antibodies faster than wait for a Histonet response (which might not be accurate anyway). It could save you your personal time as well as the time of people who read HISTONET. Laszlo Komuves FROM: L?szl? G. Komuves PhD, Senior Principal Scientist, Manager, Microscopy Core Laboratory CORGENTECH, Inc., 650 Gateway Blvd., South San Francisco, CA 94080 Phone: (650) 246-6905, Fax: (650) 624-7540, E-mail: komuves@corgentech.com BTW, I have not gotten and not going to accept any financial or non-financial compensation from any companies I mentioned. Furthermore, I am not associated with these companies (apart from being a user/customer). From lori.langiano <@t> medtronic.com Wed Jan 26 19:44:20 2005 From: lori.langiano <@t> medtronic.com (Langiano, Lori, MS, HT) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] bubbles around tissue in MMA Message-ID: <52E5C33DF6727A448293BEE0B19D01DF4E7A3F@STSM1BMSGM01> Hello all, this is my first query to the histonet. I recently started embedding tissues in MMA using the Technovit 9100 kit. I will mostly be embedding stented vessels. I have had problems with bubbles forming around the tissue during polymerization. I have not tried destabilizing the monomer yet, but did try using a vacuum before placing the sealed samples at 4 degrees C in a dessicator. Any ideas you have would be welcome. I did not find anything on this in the archives. Also, I would like to know what people use for embedding molds. Thanks, Lori From jkiernan <@t> uwo.ca Thu Jan 27 00:22:38 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] Luxol Fast Blue tissue damage References: <200501262222.j0QMMtVi028958@casbah.it.northwestern.edu> Message-ID: <41F888AE.A06418A9@uwo.ca> You have provided a superb description of ice crystal holes in CNS tissue. This is due to slow freezing of the specimen, which must be prevented. An abundance of advice is available on the Web, and it's all been there in printed books for more than 40 years. Your staining method includes some crazy steps. Have you looked up the method in a book? There are various modifications of the original method, and the mechanism of myelin staining by luxol dyes is quite well understood. Your 'regular large "holes" in my tissue' do not originate "after staining" for myelin. The empty spaces were formed when the specimens were too slowly frozen. John Kiernan London, Canada. ________________________________________________ Mathew DeGutes wrote: > > Hey all any help is appreciated. I am doing some myelin staining with luxol fast blue and have > found far too often that I get regular large "holes" in my tissue after staining. My basic > protocol is this: > > -Cryosections on slides rinse in ddH20 then PBS then 70% ethanol > -Place in .1% Luxol Fast Blue in 95% Ethanol and .05% Acetic Acid over night at 55C > -Remove from LFB and wash in ddH20 then PBS > -Differentiate in .05% Lithium Carbonate (usually about 30 sec) > -Wash in 70% ethanol and then counterstain or dehydrate and coverslip > > The actual staining looks pretty good, but I just get these Round white holes as if something > bubbled up there and destroyed the tissue. Nothing is boiling so I am curious if anyone else has > run into this problem and what might ameliorate it. Thanks again. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rentonlf <@t> bru.wits.ac.za Thu Jan 27 03:31:56 2005 From: rentonlf <@t> bru.wits.ac.za (renton louise mrs) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] bubbles around tissue in MMA Message-ID: <1106818316.8e722d7crentonlf@bru.wits.ac.za> Hi Lori, We too have experienced this problem, and there may be several expalantions: 1.In large, imperfectly infiltrated specimens...you do not mention a size, but I assume that they are reasonably small, so this might not be the case 2.Too much polymer in the mould, polymerising too fast. How long do you mix the embedding mix? I usually do 50mls no longer than 20 mins. You can try layering the polymer over the tissue and this may help Finally, if the tissue & sectioning is OK, why worry? Just carry on as usual, but if you like we can try to solve this problem, even though I am only a novice at using this resin too. Oh yes, I use polypropylene pill vials as moulds 20, 30 & 50 mls. I layer a little of the polymer on the bottom of the tube & let it set before I use the molds in order to help orient the specimen. If the specimens are tiny then BEEM capsules, Eppendorff tubes or cryotubes can be used as long as they are polyprop & not styrene. Best regards -----Original Message----- From: "Langiano, Lori, MS, HT" To: Date: Wed, 26 Jan 2005 17:44:20 -0800 Subject: [Histonet] bubbles around tissue in MMA Hello all, this is my first query to the histonet. I recently started embedding tissues in MMA using the Technovit 9100 kit. I will mostly be embedding stented vessels. I have had problems with bubbles forming around the tissue during polymerization. I have not tried destabilizing the monomer yet, but did try using a vacuum before placing the sealed samples at 4 degrees C in a dessicator. Any ideas you have would be welcome. I did not find anything on this in the archives. Also, I would like to know what people use for embedding molds. Thanks, Lori _______________________________________________ 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 marco.prunotto <@t> medecine.unige.ch Thu Jan 27 06:00:25 2005 From: marco.prunotto <@t> medecine.unige.ch (Marco Prunotto) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] immunofluorescence on paraffin sections (looking for a detailed protocol) Message-ID: <41F8D7D9.8050307@medecine.unige.ch> I hope someone can send me a good protocol to make immunofluorescence on paraffin sections. I saw that there is a an heat induced antigen retrieval method with Tris 100 mM and 5% urea.. Can someone send me a detail protocol? thanks a lot best regards ------------------------------------- Marco Prunotto, PhD Pathology Dept., University of Geneva - CH From bruyntjes <@t> voeding.tno.nl Thu Jan 27 06:26:31 2005 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:24:32 2005 Subject: [Histonet] (no subject) Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079D83@ntexch1.voeding.tno.nl> Hi all Is anyone of you familiar with an antibody directed against low or high affinity IgE receptor for rat tissue. Some time ago I found a commercial available antibody directed against low aff. IgE, but when I ordered it, the production of this antibody was finished. Joost Bruijntjes TNO Nutrition and Food Research Zeist The Netherlands This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From david.martinez <@t> dbasp.uhu.es Thu Jan 27 06:36:58 2005 From: david.martinez <@t> dbasp.uhu.es (=?iso-8859-1?Q?David_Mart=EDnez?=) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] =?iso-8859-1?q?Timm=B4s_Stain?= Message-ID: <002201c5046c$e4cee2f0$289f10ac@BCdavid> Hi, my name is David and Im a PhD student we are attempting to develop Timm's protocol in plant tissue . Now we are using the protocol posted in histonet for Greg Dobbin, but the problem is that we need a counterstained. Somebody use a counterstained for vegetal tissues after Timms protocol application? Thanks David Mart?nez ------------------------------------------------------------------------------------ Aviso del Servicio de Inform?tica y Comunicaciones (Universidad de Huelva) ------------------------------------------------------------------------------------ Se ha producido un cambio en la marcaci?n exterior de los n?meros de tel?fono de la Universidad de Huelva, que consiste simplemente en el cuarto d?gito: antes era un "0" y ahora es un "2". Por tanto, para llamadas desde el exterior, todos los n?meros han pasado de ser 95901xxxx a ser 95921xxxx. ------------------------------------------------------------------------------------ From antje.marcantonio <@t> pharma.novartis.com Thu Jan 27 06:59:02 2005 From: antje.marcantonio <@t> pharma.novartis.com (antje.marcantonio@pharma.novartis.com) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] immunofluorescence on paraffin sections (looking for a detailed protocol) Message-ID: Hi Marco the protocol depends on the antigen your are looking for , therefore I cannot give you a ready protocol. Not all antibodies require antigen retrieval to react on paraffin sections. So tell us more about the antigen then you'll have a chance to get more help and if you are lucky even a detailed protocol. Cheers, Antje Marcantonio Novartis Pharma AG BU Transplantation Research Basel, Switzerland Tel. +41 61 324 6730 antje.marcantonio@pharma.novartis.com From gcallis <@t> montana.edu Thu Jan 27 09:22:09 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Re: bubbles around tissue in MMA In-Reply-To: <52E5C33DF6727A448293BEE0B19D01DF4E7A3F@STSM1BMSGM01> References: <52E5C33DF6727A448293BEE0B19D01DF4E7A3F@STSM1BMSGM01> Message-ID: <6.0.0.22.1.20050127080356.01b2a938@gemini.msu.montana.edu> Lori, Bubbles often indicate there is rapid polymerization at too high a temperature. If you tissue is properly infiltrated, embedding can be in polypropylene vials (your stents are probably small) or some other polypropylene tube with a cap on it. I have even used conical 15 ml centrifuge tubes - you can cut the end off, push block out. Some find vials or tubes that have o-ring design within the cap - for a good seal. All major vendors have these - shipping tubes or storage vials. An open embedding mold permits excessive monomer evaporation, you get funky, hard to cut blocks since you have changed the ratio of your MMA components. Use a sealed tube, scintillation vials have been used. It is nice to have a grinder to shape a block prior to cutting and get extra plastic trimmed away down to sample to save knives and time. We tried Peel away molds but the blocks were horrible, since they were open at the top during polymerization. You really don''t need to pull a vacuum before or during polymerization. If the tissue is perfectly infiltrated, the MMA around it will polymerize along with what is in the tissue. Also, you can make a prepolymerized layer of MMA on the bottom of a tube, lay the infiltrated tissue on top of that and add your embedding MMA, the nonpolymerized monomer forms an interface with the prepolymerized layer and actually aids in polymerization, a start so to specak. Smooth, clear, bubble free polymerization occurs. This worked very well for us many times. Make sure the temperature at polymerization IS not too warm. We had bubbles if we increased the temperature to 40C, although waterbath polymerization at 37C overnight was trouble free. We made prepolymerized layers from the last infiltration mixture rather than dispose of that. Just fill tubes, tightly cap, stack them and let them sit around to polymerize. Nice way to dispose of waste MMA and have ready made prepolymerized layers. It also aids in having a flat faced block in the end. After embedding let your samples sit at RT in a hood, somehow this "rest" seems to allow bubble free polymerization - then go to a 37C waterbath, DO NOT use an incubator. Incubators do not have even heating due to prevailing air currents and you can have bubble trouble - big time. At 06:44 PM 1/26/2005, you wrote: >Hello all, this is my first query to the histonet. I recently started >embedding tissues in MMA using the Technovit 9100 kit. I will mostly be >embedding stented vessels. I have had problems with bubbles forming around >the tissue during polymerization. I have not tried destabilizing the >monomer yet, but did try using a vacuum before placing the sealed samples >at 4 degrees C in a dessicator. Any ideas you have would be welcome. I did >not find anything on this in the archives. Also, I would like to know what >people use for embedding molds. Thanks, >Lori > >_______________________________________________ >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 catherine.scott <@t> mpiresearch.com Thu Jan 27 10:20:45 2005 From: catherine.scott <@t> mpiresearch.com (Catherine Scott) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] RE:Job Opportunity (Catherine Scott) Message-ID: We (MPI Research) currently have an opening for Associate Scientist in our plastics Lab. This person would be responsible for the day-to-day operations in our newly built Plastics Lab. Candidate must have 5+ years of histology experience along with (ASCP) HT/ HTL certification. Candidate must have experience working with GMA, MMA, cardiac stents, undecalcified bone, histomorphometry and special stains. The successful candidate must be a self-starter, have leadership qualities and troubleshooting abilities. If interested please contact me a Catherine.scott@mpiresearch.com, Also, for more information please check out our web site at mpiresearch.com Thank you, Cathy Catherine L Scott, B.A., HT (ASCP), ALAT Associate Director, Pathology Services 54943 North Main Street Mattawan, MI 49071-9399 USA Telephone: 269.668-3336 ext. 1218 Email:Catherine.scott@mpiresearch.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, January 26, 2005 1:15 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 14, Issue 33 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. cd15 and morphology on bone marrows and lymph nodes (Van Eyck, Deb) 2. reagent pricing (Jeff Birkner) 3. RE: reagent pricing (Jeff Birkner) 4. RE: reagent pricing (GUTIERREZ, JUAN) 5. Re: Ache staining querry (John Kiernan) 6. Rank-L (Patty Lott) 7. Job opportunity central PA (Linda Hartman) 8. Freezing Biopsies (Amos Brooks) 9. Re: cd15 and morphology on bone marrows and lymph nodes (Andi Kappeler) 10. Eluting Solution - Glycin-HCl (Melanie Black) 11. RE: Fluorescence staining (C.M. van der Loos) 12. testing connection (Carl Hobbs) 13. subscribe to the histonet please, thanks (Wayne Holland) 14. Hairy Cell Leukemia Spleen Tissue (Barnhart, Tammy) 15. Attn: Roxanne Soto & Emily Smith (Greg Dobbin) 16. Manual fixation/processing of reconstructed skin cultures grown on polycarbonate filters (Colin Smith) 17. Re: Freezing Biopsies (Kathleen Spencer) 18. Re: cd15 and morphology on bone marrows and lymph nodes (Andi Kappeler) 19. RMSF (KarBieber@aol.com) 20. Re: Freezing Biopsies (Gayle Callis) 21. Re: RE: Fluorescence staining (Gayle Callis) 22. Re: Freezing Biopsies (ajennings@unmc.edu) 23. RE: Manual fixation/processing of reconstructed skincultures grown on polycarbonate filters (Elizabeth Chlipala) 24. mouse eye lens (Jarrin Miguel) ---------------------------------------------------------------------- Message: 1 Date: Tue, 25 Jan 2005 12:59:04 -0600 From: "Van Eyck, Deb" Subject: [Histonet] cd15 and morphology on bone marrows and lymph nodes To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain Hi all, 1.I would really appreciate some replies to a couple of questions. One is CD15; I thought I had a pretty good antibody going, but it is still problematic in some cases. I would like to know what others are using consistently with heme path approval. The same goes for kappa and lambda and clonality. 2. The other issue is handling bone marrows and lymph nodes---we have recently gotten rid of B-5 and we use NBF as our main fixative. I would like some replies in regards to what is the best fixation/decal situation for morphology purposes (does anyone feel that they have reached the b-5 nirvana state for good morphology) and still maintain immuno staining???? Times and protocols and products would be great. Thanks----Deb Van Eyck-Anatomic Path Waukesha Memorial Hospital 725 American Ave. Waukesha,WI 53188 262-928-2112 fax:262-928-4849 This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. ------------------------------ Message: 2 Date: Tue, 25 Jan 2005 13:17:01 -0600 From: Jeff Birkner Subject: [Histonet] reagent pricing To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain What is everyone paying for reagent alcohol, xylene and formalin? We are paying the following per gallon, not including shipping: Reagent alcohol $15.00 Xylene $28.00 10% NB Formalin $5.45 Jeff Birkner, CT(ASCP) Pathology Section Manager Collaborative Laboratory Services, LLC 1005 Pennsylvania Ave. Ottumwa, IA 52501 ------------------------------ Message: 3 Date: Tue, 25 Jan 2005 13:56:47 -0600 From: Jeff Birkner Subject: [Histonet] RE: reagent pricing To: "'histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain I will put the responses in an excel spreadsheet and share them with anyone that wants them. _____ From: Jeff Birkner Sent: Tuesday, January 25, 2005 1:17 PM To: 'histonet@lists.utsouthwestern.edu' Subject: reagent pricing What is everyone paying for reagent alcohol, xylene and formalin? We are paying the following per gallon, not including shipping: Reagent alcohol $15.00 Xylene $28.00 10% NB Formalin $5.45 Jeff Birkner, CT(ASCP) Pathology Section Manager Collaborative Laboratory Services, LLC 1005 Pennsylvania Ave. Ottumwa, IA 52501 ------------------------------ Message: 4 Date: Tue, 25 Jan 2005 14:14:17 -0600 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] reagent pricing To: "Jeff Birkner" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Reagent alcohol 8.73/gal Xylene 7.85/gal Formalin 2.92/gal Say Hi to Radar for me. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jeff Birkner Sent: Tuesday, January 25, 2005 1:17 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] reagent pricing What is everyone paying for reagent alcohol, xylene and formalin? We are paying the following per gallon, not including shipping: Reagent alcohol $15.00 Xylene $28.00 10% NB Formalin $5.45 Jeff Birkner, CT(ASCP) Pathology Section Manager Collaborative Laboratory Services, LLC 1005 Pennsylvania Ave. Ottumwa, IA 52501 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Tue, 25 Jan 2005 15:49:42 -0500 From: John Kiernan Subject: Re: [Histonet] Ache staining querry To: Arvind Pundir Cc: histonet@lists.utsouthwestern.edu Message-ID: <41F6B0E6.5B99D9E2@uwo.ca> Content-Type: text/plain; charset=us-ascii Assuming "Ache staining" is acetylcholinesterase activity histochemistry, not a painful bruise: The surface of the specimen will have been fixed more than the inside. AChE activity resists short fixation in formaldehyde (such as overnight). Perhaps the outside of the block has had more than is good for the enzyme. In small specimens of animal tissues, both the choline esterases (and other esterases) are well preserved by overnight fixation in formaldehyde. Human brains need much longer because of the large size, and this can be expected to wreck enzyme activities. You might still be able to stain the AChE protein with an immunohistochemical method. -- ------------------------------- 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 _______________________________ Arvind Pundir wrote: > > i do Ache staining frequently on human brain sections, but facing problem that there is no staining at the edges of my section can anyone sort what it may be due to > > > > thanks in advance for any suggestion or advice > > > > Arvind Singh Pundir > > National Brain Research Centre > > INDIA > > ------------------------------------------------------------ > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Tue, 25 Jan 2005 16:30:45 -0600 From: Patty Lott Subject: [Histonet] Rank-L To: 'histonet' Message-ID: <85F6C7A1330E794DB8540AFD001CC77E05328DA6@rosco.ortho.uab.edu> Content-Type: text/plain Does anyone know of an antibody for RANK-L to be used on FFPE mouse bones? Patty Lott, Laboratory Supervisor Orthopaedic Research Laboratory Center for Metabolic Bone Disease Laboratory University of Alabama at Birmingham LHRB B37 0007 1919 7th Ave. South Birmingham, AL 35294 (205) 934-2007 ------------------------------ Message: 7 Date: Tue, 25 Jan 2005 20:38:45 -0500 From: "Linda Hartman" Subject: [Histonet] Job opportunity central PA To: Message-ID: <002501c50347$c7335a80$1d04a245@linda47p53fn9e> Content-Type: text/plain; charset="iso-8859-1" Mount Nittany Medical Center in State College, PA has an opening for a full time histology technician. The applicant will be responsible for routine histology, frozen sections, special staining and IHC. Mount Nittany Medical Center is a growing regional medical center located mid-way between Philadelphia and Pittsburgh. State College is the home of the main campus of Penn State University. Please check out our website at www.mountnittany.org Linda Hartman HT(ASCP) Histology section head Mount Nittany Medical Center State College, PA 16803 Phone: 814-234-6117 email: lhartman@mountnittany.org ------------------------------ Message: 8 Date: Tue, 25 Jan 2005 21:07:30 -0500 From: Amos Brooks Subject: [Histonet] Freezing Biopsies To: histonet@lists.utsouthwestern.edu Message-ID: <6.2.0.14.0.20050125210242.01da7198@pop.earthlink.net> Content-Type: text/plain; charset="us-ascii"; format=flowed Hi, I'm snap freezing kidney biopsies in OCT using isopentane that is chilled with liquid nitrogen. I'm having a problem where the area that the biopsy is gets raised up above the surface of the block. I'm not sure what is going wrong and more importantly what to do about it. Has anyone had similar problems and how did you solve it? Thanks, Amos Brooks ------------------------------ Message: 9 Date: Wed, 26 Jan 2005 07:52:43 +0100 From: "Andi Kappeler" Subject: Re: [Histonet] cd15 and morphology on bone marrows and lymph nodes To: "Histonet" , "Van Eyck, Deb" Message-ID: <002401c50373$a3254be0$27955c82@patho.unibe.ch> Content-Type: text/plain; charset="Windows-1252" Hi Deb We use mo-a-CD15, clone MMA, Becton-Dickinson 347420. Working conc. is 2.5 ug Ig/ml, pretreatment is HIER (citrate) in pressure cooker. The 'secret' with this clone (and most other commercially available CD15 mAbs, such as C3D-1, BY87, 80H5, and more) is, that it is of Ig class IgM (and not IgG1, IgG2a, as most monoclonals that you commonly use). While most secondary antibodies show some cross-reactivity with mAbs of Ig class IgM, you have to use a mouse-IgM specific secondary antibody to get optimal results (e.g. DakoCytomation E 0465, rb-a-mo IgM/Biotin). This applies also to a couple of other mAbs: they will work with 'regular' secondary antibodies, however, they will work better with an IgM-specific secondary antibody. Therefore it is always useful to check the SpecSheet for the Ig-class of a given mAb. If you are using a visualization system that comes from the supplier of your staining machine, you may have a problem, depending on the stainer. Some of them are obviously very reluctant to let you use secondary reagents of your choice... Then it is good to know, that immunos can be done manually ... Hope this helps. Good luck. Andi Kappeler Insitute of Pathology, University of Bern, Switzerland ----- Original Message ----- From: "Van Eyck, Deb" To: Sent: Tuesday, January 25, 2005 7:59 PM Subject: [Histonet] cd15 and morphology on bone marrows and lymph nodes > > Hi all, > 1.I would really appreciate some replies to a couple of questions. One is > CD15; I thought I had a pretty good antibody going, but it is still > problematic in some cases. I would like to know what others are using > consistently with heme path approval. The same goes for kappa and lambda > and clonality. > > > 2. The other issue is handling bone marrows and lymph nodes---we have > recently gotten rid of B-5 and we use NBF as our main fixative. I would > like some replies in regards to what is the best fixation/decal situation > for morphology purposes (does anyone feel that they have reached the b-5 > nirvana state for good morphology) and still maintain immuno staining???? > Times and protocols and products would be great. > > > Thanks----Deb Van Eyck-Anatomic Path > Waukesha Memorial Hospital > 725 American Ave. > Waukesha,WI 53188 > 262-928-2112 > fax:262-928-4849 > > > > This information is confidential and intended solely for the use of the > individual or entity to whom it is addressed. > If you have received this email in error please notify the sender or our > Customer Support Center at (262) 928-2777. > > We have scanned this email and its attachments for malicious content. > However, the recipient should check this email and any attachments for the > presence of viruses. > ProHealth Care accepts no liability for any damage caused by any virus > transmitted by this email. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 10 Date: Wed, 26 Jan 2005 09:21:12 +0200 From: Melanie Black Subject: [Histonet] Eluting Solution - Glycin-HCl To: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Hi There Can anybody give me a method for making up Glycin-HCl(0.1M, pH 2.2) to be used an a blocking buffer in double immuno staining??? 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 ------------------------------ Message: 11 Date: Wed, 26 Jan 2005 08:52:21 +0100 From: "C.M. van der Loos" Subject: [Histonet] RE: Fluorescence staining To: histonet@lists.utsouthwestern.edu Message-ID: <12c276c12c2991.12c299112c276c@amc.uva.nl> Content-Type: text/plain; charset="us-ascii" Dear Eva, To my knowledge most of the traditional fluorochromes like FITC, rhodamine, doesn't stand dehydration at all. So in your case it fully depends on what type of fluorochrome you end up with after the TSA procedure. If you end up with FITC, the use of VectaShield ([1]www.vectorlabs.com) for mounting is highly recommended for the prevention of photobleaching. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: [2]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From Eva C Anderson Date Tue, 25 Jan 2005 11:10:41 -0500 To histonet@lists.utsouthwestern.edu Subject [Histonet] Fluorescence staining Hi, We are looking into trying some Fluorescence staining. We want to try a TSA Fluorescence System from PerkinElmer. The IHC protocol does however leave me with some questions. We always dehydrate our samples after staining. The protocol however doesn't mention this. It only says that the samples can be counterstained and mounted after incubation with the Fluorophore Tyramide. Should we dehydrate and is there a particular mounting media we should use to prolong the time that fluorescence can be detected? Any help would be greatly appreciated. Thanks in advance for all your help, Eva Georgetown University Research Assistant References 1. http://www.vectorlabs.com/ 2. mailto:c.m.vanderloos@amc.uva.nl ------------------------------ Message: 12 Date: Wed, 26 Jan 2005 08:09:43 -0000 From: "Carl Hobbs" Subject: [Histonet] testing connection To: Message-ID: <000301c5037e$646dfea0$e8345c9f@Carlos> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original My emails keep getting bounced ------------------------------ Message: 13 Date: Wed, 26 Jan 2005 07:10:50 -0500 From: "Wayne Holland" Subject: [Histonet] subscribe to the histonet please, thanks To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ ------------------------------ Message: 14 Date: Wed, 26 Jan 2005 07:29:02 -0600 From: "Barnhart, Tammy" Subject: [Histonet] Hairy Cell Leukemia Spleen Tissue To: "Histonet (E-mail)" Message-ID: <1779904B5E82D511914C00D0B793339205BFD8B1@exchangent> Content-Type: text/plain; charset="iso-8859-1" I am in need of the above control tissue. We have not had a case here that we can find and would like to use it as a control for our TRAcP antibody. Willing to pay, hopefully not too much. Or, trade for other tissue :) Thanks.... Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND tbarnhart@primecare.org 701-530-6730 Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. ------------------------------ Message: 15 Date: Wed, 26 Jan 2005 09:48:59 ADT From: "Greg Dobbin" Subject: [Histonet] Attn: Roxanne Soto & Emily Smith To: histonet@lists.utsouthwestern.edu Message-ID: <41F7678B.22027.1B921F@localhost> Content-Type: text/plain; charset=US-ASCII Roxanne and Emily, I am getting planning underway again for the Histonet Outpost. I sent out an e-mail to our `ad hoc' committee members yesterday and your e-mails bounced. Can you confirm that your e-mail addresses (below) are correct and if not, provide the proper one so I can forward you the e-mail again as well as all the replies I received to it. Thanks. Greg ESmith@CuraGen.com GREYTRUNK@aol.com ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Research Technologist, Pathology Lab, Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. ------------------------------ Message: 16 Date: Wed, 26 Jan 2005 15:07:05 -0000 From: "Colin Smith" Subject: [Histonet] Manual fixation/processing of reconstructed skin cultures grown on polycarbonate filters To: Message-ID: <3E985CD1D7CA89499D1902BC3202AF3CC1A75E@uk-res-srv-03.thorspecialities.c om> Content-Type: text/plain; charset="us-ascii" Can anyone help? I am currently setting up a basic histology system for analysing reconstructed tissues grown in culture on polycarbonate filters, similar to Mattek's Epiderm. We currently have begged borrowed or stolen, equipment-wise: two water baths (ambient - 100C), a circulating warm air oven, a rocking microtome and cold blocks, along with glass staining jars/racks, forceps etc. At the moment, equipment such as a wax dispenser, purpose built cooling plates etc are out of the question until I can show that we can show that the model we are growing is validated, which will include immunohistochemistry of cytokeratin and cell adhesion markers etc. Thus we are in the catch 22 situation of having to prove something without the equipment needed to do the job, in order to justify buying the equipment!! But thats accountants for you, go figure. Anyway, can anyone give any useful pointers/protocols for fixing/processing/embedding/sectioning such tissues using such primative equipment Any help, no matter how trivial will be greatfully received. Many Thanks in Advance, Colin Smith BSc CBiol MIBiol Eurotox Registered Toxicologist In Vitro Toxicologist Thor Specialities UK Ltd Wincham Avenue, Northwich CW9 6GB Tel: +44 1606 818873 Fax: +44 1606 818801 colins@thor.uk.com The information contained in this e-mail may be confidential and may also be legally privileged. It is intended solely for the addressee(s). If you are not the intended recipient then please delete forthwith. Views expressed in this email are views of the individual and are not necessarily the views of this company. ************************************************************************ ************************* e-mail scanned for Viruses by Thor Specialities (UK) Limited using Gfi Mail Security ************************************************************************ ************************* ------------------------------ Message: 17 Date: Wed, 26 Jan 2005 09:39:59 -0600 From: Kathleen Spencer Subject: Re: [Histonet] Freezing Biopsies To: Amos Brooks Cc: histonet@lists.utsouthwestern.edu Message-ID: <89982A33-6FB0-11D9-8BD6-000393967904@utmem.edu> Content-Type: text/plain; charset=US-ASCII; format=flowed Amos, The way we did this was to place the biopsy in a plastic mold, add OCT, grab the huge forceps and plunge it into liquid nitrogen and count to 5 or 10, I can't remember, but it always worked fine. We did not use isopentane. The surface of the block was always nice and smooth. It had to be! The biopsies were so tiny. What you can do is get a fresh mold, put a little OCT in the bottom, put the frozen block in and spray it a little with cryo spray. You should now have a smooth surface for sectioning. Good Luck! Kathleen Spencer HT (ASCP) Lab Manager/LCM Supervisor UTHSC On Jan 25, 2005, at 8:07 PM, Amos Brooks wrote: > Hi, > I'm snap freezing kidney biopsies in OCT using isopentane that is > chilled with liquid nitrogen. I'm having a problem where the area that > the biopsy is gets raised up above the surface of the block. I'm not > sure what is going wrong and more importantly what to do about it. Has > anyone had similar problems and how did you solve it? > > Thanks, > Amos Brooks > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Wed, 26 Jan 2005 17:18:31 +0100 From: "Andi Kappeler" Subject: Re: [Histonet] cd15 and morphology on bone marrows and lymph nodes To: "Histonet" Message-ID: <006301c503c2$ad285ed0$27955c82@patho.unibe.ch> Content-Type: text/plain; charset="iso-8859-1" We use the rb-a-mo IgM/Biotin in stead of our usual anti-mouse Ig secondary antibody (which is gt-a-mo Ig/Biotin). As we still stain manually (yes that exists...!) this is no problem. Adding the rb-a-mo IgM to your usual secondary is theoretically possible, however if your secondary is a 'multi link' reagent (anti-mouse and anti-rabbit cocktail) then you should not add a rb-a-mo IgM/Biotin as it will be bound by the anti-rb component of your 'multi link' reagent. The system may still work, but I would prefer to know what I'm doing rather than to 'let something happen' inside a tube where I don't have control. If you want to add an anti-mouse IgM component to your 'multi link', make sure it does not intefere with any of the components that is already in there (e.g. use a gt-a-mo IgM/Biotin). Our protocol for CD15 looks as follows (in brief): 1. Deparaffinization --> TBS 2. Pretreatment, citrate buffer, pressure cooker, 7 min at 121 C, 1 bar; cool down, wash 3. Incubate with mo-a-CD15, clone MMA, 2.5 ug Ig/ml 4. Wash 5. Incubate with rb-a-mo IgM (DakoCytomation E 0465, 1:500) 6. Wash 7. Incubate with enzyme system (ABC/HRP, LSAB+/HRP, whatever...) 8. Wash 9. Develop with 3,3'-diaminobenzidin /H2O2 10. Rinse, counterstain, mount Best regards, Andi ----- Original Message ----- From: "Van Eyck, Deb" To: "'Andi Kappeler'" Sent: Wednesday, January 26, 2005 4:20 PM Subject: RE: [Histonet] cd15 and morphology on bone marrows and lymph nodes > thanks very much-------for the secondary system do you just use this with > another Dako product for example---are you using lsab or lsab +. I know > some people use this system and just add the IgM to the secondary--what > specifically do you do. this has been a real help Andi! > > > -----Original Message----- > > From: Andi Kappeler [SMTP:kappeler@patho.unibe.ch] > > Sent: Wednesday, January 26, 2005 12:53 AM > > To: Histonet; Van Eyck, Deb > > Subject: Re: [Histonet] cd15 and morphology on bone marrows and lymph > > nodes > > > > Hi Deb > > > > We use mo-a-CD15, clone MMA, Becton-Dickinson 347420. Working conc. is 2.5 > > ug Ig/ml, pretreatment is HIER (citrate) in pressure cooker. The 'secret' > > with this clone (and most other commercially available CD15 mAbs, such as > > C3D-1, BY87, 80H5, and more) is, that it is of Ig class IgM (and not > > IgG1, > > IgG2a, as most monoclonals that you commonly use). While most secondary > > antibodies show some cross-reactivity with mAbs of Ig class IgM, you > > have > > to use a mouse-IgM specific secondary antibody to get optimal results > > (e.g. > > DakoCytomation E 0465, rb-a-mo IgM/Biotin). This applies also to a couple > > of > > other mAbs: they will work with 'regular' secondary antibodies, however, > > they will work better with an IgM-specific secondary antibody. Therefore > > it > > is always useful to check the SpecSheet for the Ig-class of a given mAb. > > If you are using a visualization system that comes from the supplier of > > your > > staining machine, you may have a problem, depending on the stainer. Some > > of > > them are obviously very reluctant to let you use secondary reagents of > > your > > choice... Then it is good to know, that immunos can be done manually ... > > Hope this helps. Good luck. > > > > Andi Kappeler > > Insitute of Pathology, University of Bern, Switzerland > > > > > > > > ----- Original Message ----- > > From: "Van Eyck, Deb" > > To: > > Sent: Tuesday, January 25, 2005 7:59 PM > > Subject: [Histonet] cd15 and morphology on bone marrows and lymph nodes > > > > > > > > > > Hi all, > > > 1.I would really appreciate some replies to a couple of questions. One > > is > > > CD15; I thought I had a pretty good antibody going, but it is still > > > problematic in some cases. I would like to know what others are using > > > consistently with heme path approval. The same goes for kappa and > > lambda > > > and clonality. > > > > > > > > > 2. The other issue is handling bone marrows and lymph nodes---we have > > > recently gotten rid of B-5 and we use NBF as our main fixative. I would > > > like some replies in regards to what is the best fixation/decal > > situation > > > for morphology purposes (does anyone feel that they have reached the b-5 > > > nirvana state for good morphology) and still maintain immuno > > staining???? > > > Times and protocols and products would be great. > > > > > > > > > Thanks----Deb Van Eyck-Anatomic Path > > > Waukesha Memorial Hospital > > > 725 American Ave. > > > Waukesha,WI 53188 > > > 262-928-2112 > > > fax:262-928-4849 > > > > > > > > > > > > This information is confidential and intended solely for the use of the > > > individual or entity to whom it is addressed. > > > If you have received this email in error please notify the sender or our > > > Customer Support Center at (262) 928-2777. > > > > > > We have scanned this email and its attachments for malicious content. > > > However, the recipient should check this email and any attachments for > > the > > > presence of viruses. > > > ProHealth Care accepts no liability for any damage caused by any virus > > > transmitted by this email. > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > This information is confidential and intended solely for the use of the > individual or entity to whom it is addressed. > If you have received this email in error please notify the sender or our > Customer Support Center at (262) 928-2777. > > We have scanned this email and its attachments for malicious content. > However, the recipient should check this email and any attachments for the > presence of viruses. > ProHealth Care accepts no liability for any damage caused by any virus > transmitted by this email. > > ------------------------------ Message: 19 Date: Wed, 26 Jan 2005 11:23:56 EST From: KarBieber@aol.com Subject: [Histonet] RMSF To: histonet@pathology.swmed.edu Message-ID: <145.3e0c9baa.2f291e1c@aol.com> Content-Type: text/plain; charset="US-ASCII" Does anyone have a good source for an antibody for RMSF? Thanks, Karen ------------------------------ Message: 20 Date: Wed, 26 Jan 2005 09:25:16 -0700 From: Gayle Callis Subject: Re: [Histonet] Freezing Biopsies To: Amos Brooks , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050126085604.01af5e68@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Amos, We experienced this too. We solved by using Tissue Tek plastic embedding molds 10 x 10 mm size. Cut excess edges for a better heat sink around embedding well except for a small tab to hold onto. Carefully pick a mold that is not deformed already - look for the flat ones, not rounded! Tissue Tek has newer round molds that do not require trimming, very nice, tidy and FLAT! Unfortunately, other brands of plastic molds we tried had thicker plastic compared to Tissue Tek molds and blocks twisted out at freezing. Coat bottom of mold with a THIN film of OCT. Biopsy is laid out so it is as flat as possible in OCT film on bottom. Carefully, slowly - add more OCT down corner of mold or away from top of biopsy to keep it from floating up. Overfilling with OCT results in cracked blocks; fill to top of well. We tilt the beaker with cold isopentane , and introduce mold so bottom makes contact with isopentant first. When OCT starts to turn white ON BOTTOM, immerse fully -about 10 sec or so. Bottom freezing first anchors biopsy and keeps it flat, no lift up syndrome. If frozen block face has any deformity to it caused by a funky mold - just smear a thin layer of OCT over block face to fill in these depressions, etc. to have a flatter block face before trimming. Thin layer is important as we tried to avoid any excess warming of block once it is frozen solid. Have fun!! At 07:07 PM 1/25/2005, you wrote: >Hi, > I'm snap freezing kidney biopsies in OCT using isopentane that is > chilled with liquid nitrogen. I'm having a problem where the area that > the biopsy is gets raised up above the surface of the block. I'm not sure > what is going wrong and more importantly what to do about it. Has anyone > had similar problems and how did you solve it? > >Thanks, >Amos Brooks > > > >_______________________________________________ >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: 21 Date: Wed, 26 Jan 2005 09:33:09 -0700 From: Gayle Callis Subject: Re: [Histonet] RE: Fluorescence staining To: "C.M. van der Loos" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050126092847.01b583c0@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Chris and Eva, Vectashield also come in a Hard Set, so you get permanent coverslipping (no coverglass sliding around!) as does the Molecular Probes Prolong Gold ready to use. If you want a counterfluorescence marker for nuclei, and have the appropriate DAPI filter on microscope, you can buy these with DAPI which "counterstains via fluorescence" the DNA in cells. At 12:52 AM 1/26/2005, you wrote: > Dear Eva, > > To my knowledge most of the traditional fluorochromes like FITC, > rhodamine, doesn't stand dehydration at all. So in your case it fully > depends on what type of fluorochrome you end up with after the TSA > procedure. > > If you end up with FITC, the use of VectaShield > ([1]www.vectorlabs.com) for mounting is highly recommended for > the prevention of photobleaching. > > Chris van der Loos, PhD > Dept. of Pathology > Academical Medical Center M2-230 > Meibergdreef 9 > NL-1105 AZ Amsterdam > The Netherlands > > phone: +31 20 5665631 > fax: +31 20 6960389 > e-mail: [2]c.m.vanderloos@amc.uva.nl 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: 22 Date: Wed, 26 Jan 2005 10:33:22 -0600 From: ajennings@unmc.edu Subject: Re: [Histonet] Freezing Biopsies Cc: histonet@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu, Amos Brooks Message-ID: Content-Type: text/plain; charset=US-ASCII Bernice from Instrumedics I am sure can give you better insight into what use to be called "Gentle Jane" freezing. Helps with heat extraction and overall consistent freezing. It may help. I use to have the same problem with whole mouse embryos, because they were freezing from the outside in and by the time the liver froze/expanded it would pop out of the block. I know biopsies are by far smaller but it could be the same general theory with the OCT freezing from the outside around the sample and the sample having no place else to expand. It actually helped me to freeze in the lN2 vapor instead of dropping the sample into isopentane. The time difference was negligible and I didn't get that pop out peak that I believe you are referring to. The freezing occurred from the bottom of the boat to the top instead from all sides at once. It might not be appropriate for all but it has worked in my lab for many many years. just sharing a thought On Jan 25, 2005, at 8:07 PM, Amos Brooks wrote: > Hi, > I'm snap freezing kidney biopsies in OCT using isopentane that is > chilled with liquid nitrogen. I'm having a problem where the area that > the biopsy is gets raised up above the surface of the block. I'm not > sure what is going wrong and more importantly what to do about it. Has > anyone had similar problems and how did you solve it? > > Thanks, > Amos Brooks ------------------------------ Message: 23 Date: Wed, 26 Jan 2005 09:36:26 -0700 From: "Elizabeth Chlipala" Subject: RE: [Histonet] Manual fixation/processing of reconstructed skincultures grown on polycarbonate filters To: "'Colin Smith'" , Message-ID: <000001c503c5$2e345770$76d48a80@AMY> Content-Type: text/plain; charset="us-ascii" Colin We have worked quite a bit with the epioral and epigingival cultures from Mattek. We have stained with H&E, Ki-67 and Cleaved caspase 3. We processed these specimens on a tissue processor, but you will be able to do this by hand. I'm not sure of the size of your culture wells, but we used an 8mm biopsy punch to remove the constructs from the wells, processed the construct whole between two biopsy pads and bisected with a razor blade prior to embedding on end. As far as fixation 10% NBF will work. The wells were labeled on the side and the entire well was placed in a 50 ml conical tube with fixative. After adequate fixation the specimens were grossed, processed and embedded into paraffin. We processed during the day (not overnight) at 15 minutes per station. We sectioned at 4 microns and stained with both H&E and various Immunohistochemical stains, I can send you images if you are interested. If you need more info, give me a call or e-mail me back. Also we are writing an article that will be in the spring edition of Histologic that basically covers the methods (both processing and analysis) we used to assess proliferation in these in vitro tissue models. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Colin Smith Sent: Wednesday, January 26, 2005 8:07 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Manual fixation/processing of reconstructed skincultures grown on polycarbonate filters Can anyone help? I am currently setting up a basic histology system for analysing reconstructed tissues grown in culture on polycarbonate filters, similar to Mattek's Epiderm. We currently have begged borrowed or stolen, equipment-wise: two water baths (ambient - 100C), a circulating warm air oven, a rocking microtome and cold blocks, along with glass staining jars/racks, forceps etc. At the moment, equipment such as a wax dispenser, purpose built cooling plates etc are out of the question until I can show that we can show that the model we are growing is validated, which will include immunohistochemistry of cytokeratin and cell adhesion markers etc. Thus we are in the catch 22 situation of having to prove something without the equipment needed to do the job, in order to justify buying the equipment!! But thats accountants for you, go figure. Anyway, can anyone give any useful pointers/protocols for fixing/processing/embedding/sectioning such tissues using such primative equipment Any help, no matter how trivial will be greatfully received. Many Thanks in Advance, Colin Smith BSc CBiol MIBiol Eurotox Registered Toxicologist In Vitro Toxicologist Thor Specialities UK Ltd Wincham Avenue, Northwich CW9 6GB Tel: +44 1606 818873 Fax: +44 1606 818801 colins@thor.uk.com The information contained in this e-mail may be confidential and may also be legally privileged. It is intended solely for the addressee(s). If you are not the intended recipient then please delete forthwith. Views expressed in this email are views of the individual and are not necessarily the views of this company. ************************************************************************ ************************* e-mail scanned for Viruses by Thor Specialities (UK) Limited using Gfi Mail Security ************************************************************************ ************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 24 Date: Wed, 26 Jan 2005 09:48:38 -0800 (PST) From: Jarrin Miguel Subject: [Histonet] mouse eye lens To: histonet@lists.utsouthwestern.edu Message-ID: <20050126174838.64967.qmail@web41901.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Dear all, Let me introduce myself. My name is Miguel and I am PhD student. I am considering to carry out immunochemistry and in situ hybridization studies in parafin sections of eye lens in adult mouse. I would like receive futher information about the best fixative that use. I have certain experience with Paraformaldehide, but I have been advised about Davison's fixative and Penfix. I never have heared about Penfix before. Somebody has protocols or receipt for the penfix? Thanking in advance for your help Miguel --------------------------------- 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 End of Histonet Digest, Vol 14, Issue 33 **************************************** ****************************************************************************** Visit the MPI Website at http://www.mpiresearch.com Confidentiality Notice: "By accessing this e-mail communication, the intended recipient hereby consents to the transmittal and receipt of confidential information via electronicmedium. This e-mail is strictly confidential and intended solely for the addressee. If you are not the intended recipient you must not use, disclose, or copy this transmission. If you have received this message in error, please contact the sender and delete the material from any computer, disc drive, diskette, or other storage device or media. This e-mail is not intended to impose, nor shall it be construed as imposing, any legally binding obligation upon MPI Research, Inc., or any of its subsidiaries or affiliated companies. Neither MPI Research, Inc. nor any of its subsidiaries or affiliated companies gives any representation or warranty as to the accuracy or completeness of the contents of this e-mail. MPI Research, Inc. shall not be held liable to any person resulting from the use of any information contained in this e-mail, and shall not be liable to any person who acts or omits to do anything in reliance upon receipt of this e-mail. MPI Research, Inc. hereby expressly disclaims any and all liability for any viruses or data corrupting contents which may result from recipient’s access or use of this e-mail or its contents." ******************************************************************************* MPI Research, Inc. - (269) 668-3336 From pruegg <@t> ihctech.net Thu Jan 27 10:51:03 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] reagent pricing In-Reply-To: Message-ID: <200501271651.j0RGoxpL019929@chip.viawest.net> StatLabs has the best prices on reagents that I have found. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of GUTIERREZ, JUAN Sent: Tuesday, January 25, 2005 1:14 PM To: Jeff Birkner; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] reagent pricing Reagent alcohol 8.73/gal Xylene 7.85/gal Formalin 2.92/gal Say Hi to Radar for me. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jeff Birkner Sent: Tuesday, January 25, 2005 1:17 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] reagent pricing What is everyone paying for reagent alcohol, xylene and formalin? We are paying the following per gallon, not including shipping: Reagent alcohol $15.00 Xylene $28.00 10% NB Formalin $5.45 Jeff Birkner, CT(ASCP) Pathology Section Manager Collaborative Laboratory Services, LLC 1005 Pennsylvania Ave. Ottumwa, IA 52501 _______________________________________________ Histonet mailing list Histonet@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 rocan <@t> mac.com Thu Jan 27 11:15:54 2005 From: rocan <@t> mac.com (Rocan) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] immunofluorescence on paraffin sections (looking for a detailed protocol) In-Reply-To: <41F8D7D9.8050307@medecine.unige.ch> References: <41F8D7D9.8050307@medecine.unige.ch> Message-ID: <1A5E7A72-7087-11D9-9735-000A9589219E@mac.com> Marco, In my hands, immunofluorescence on paraffin is a very frustrating and unrewarding task. I am sorry to have to say this. Fixatives that crosslink tissue (para- and formaldehyde) cause massive collagen crosslinking. Collagen is inherently highly auto-fluorescent. If the bundles of collagen are cross-linked the autofluorescence is increased to a point in which it strongly interferes with most fluorochromes used for immunofluorescence. There may be some ways to reduce the autofluorescence of cross-linked tissue and I am sure you would be receiving the best advice in this forum. However, I thought it would be important to mention the major setbacks of the technique just incase you may have not been aware of this. When possible, use frozen section of fresh unfixed tissue or even better, whole explants if and when this possibility exists (tissue slices, retinas,peritoneum, etc) ----- Dr.Rocio Sierra-Honigmann Director Engineered Wound Repair Laboratory Cedars Sinai Research Institute Davis 1091 310-423-1882 Honigmannr@cshs.org On Jan 27, 2005, at 4:00 AM, Marco Prunotto wrote: > I hope someone can send me a good protocol to make immunofluorescence > on paraffin sections. > I saw that there is a an heat induced antigen retrieval method with > Tris 100 mM and 5% urea.. > Can someone send me a detail protocol? > > thanks a lot > > best regards > > > ------------------------------------- > Marco Prunotto, PhD > Pathology Dept., University of Geneva - CH > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hymclab <@t> hyhc.com Thu Jan 27 12:46:33 2005 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] reagent pricing Message-ID: SURGIPATH has my vote. We pay less than those prices listed below. Dawn -----Original Message----- From: Patsy Ruegg [mailto:pruegg@ihctech.net] Sent: Thursday, January 27, 2005 10:51 AM To: 'GUTIERREZ, JUAN'; 'Jeff Birkner'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] reagent pricing StatLabs has the best prices on reagents that I have found. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of GUTIERREZ, JUAN Sent: Tuesday, January 25, 2005 1:14 PM To: Jeff Birkner; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] reagent pricing Reagent alcohol 8.73/gal Xylene 7.85/gal Formalin 2.92/gal Say Hi to Radar for me. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jeff Birkner Sent: Tuesday, January 25, 2005 1:17 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] reagent pricing What is everyone paying for reagent alcohol, xylene and formalin? We are paying the following per gallon, not including shipping: Reagent alcohol $15.00 Xylene $28.00 10% NB Formalin $5.45 Jeff Birkner, CT(ASCP) Pathology Section Manager Collaborative Laboratory Services, LLC 1005 Pennsylvania Ave. Ottumwa, IA 52501 _______________________________________________ Histonet mailing list Histonet@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 Wanda.Smith <@t> HCAhealthcare.com Thu Jan 27 12:54:27 2005 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Fri Sep 16 15:24:33 2005 Subject: **SPAM** [Histonet] reagent pricing Message-ID: <8784A3EE34199644AF60DBE517F5B0A60A8A5786@louex04.lou.medcity.net> Jeff, We pay the following: Reagent alcohol $10.79/gal Xylene $9.47/gal 10% NBF $25.70/per 5 gal cube 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: Jeff Birkner [mailto:jbirkner@colabserv.com] Sent: Tuesday, January 25, 2005 2:17 PM To: 'histonet@lists.utsouthwestern.edu' Subject: **SPAM** [Histonet] reagent pricing What is everyone paying for reagent alcohol, xylene and formalin? We are paying the following per gallon, not including shipping: Reagent alcohol $15.00 Xylene $28.00 10% NB Formalin $5.45 Jeff Birkner, CT(ASCP) Pathology Section Manager Collaborative Laboratory Services, LLC 1005 Pennsylvania Ave. Ottumwa, IA 52501 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Dorothy.L.Webb <@t> HealthPartners.Com Thu Jan 27 14:34:23 2005 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] processing help Message-ID: I have a dermatopathologist who complained to me that his slides on the skins we short processed yesterday looked very eosinophilic and the collagen was separating out. Does anyone out there have any ideas what the problem could be?? We changed nothing in our processor from one day to the next nor any of the times. OUr routine tissues from today looked great! What could possibly be the problem when a pathologist comes across with this problem?? HELP!! And Thanks!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From sharon.osborn <@t> dnax.org Thu Jan 27 15:47:32 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Shandon Excelsior processing center Message-ID: <29B25753F6B1D51196110002A589D44402397EB1@PALMSG30.us.schp.com> Histonetters, The organizational controller and my direct supervisor have requested I ask for this information. Question: What is your experience with the ThermoShandon Excelsior processing center for reliability? Do you find it beneficial or necessary to have the service contract and net monitoring? We have had one for a year and are told the service contract and net monitoring is beneficial/necessary to receive the upgrades to the unit and for monitoring problems. How many of you have a service contract on these for 1 or 2 years and do not renew it? In our company, these service contracts are reviewed legally and fiscally so it is important I research information to support whatever decision is recommended. Thanks for your input. 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 Bultema.Brent <@t> mayo.edu Thu Jan 27 15:51:03 2005 From: Bultema.Brent <@t> mayo.edu (Bultema, Brent T.) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Histopathology Supervisor opportunity - Mayo Clinic Message-ID: <3CE454ED960EEB4387BDE95E401776A801E678D5@excsrv18.mayo.edu> HISTOPATHOLOGY SUPERVISOR Mayo Clinic in Rochester, MN, currently has an opportunity available for a Histopathology Supervisor, Histology Lab. The selected candidate will manage both resources and personnel in his/her particular area, ensuring sound fiscal leadership, quality service, process improvements and quality performance. Must be able to develop, evaluate and motivate staff members. A bachelor's degree - preferably in Medical Technology, management or a health-or science-related field - prefer HTL (ASCP) registration or eligible (30 semester hours of biology and chemistry and 1 year histology lab experience). Other qualifications: 4+ years' clinical lab medicine experience, excellent communication and interpersonal skills, and computer proficiency. Mayo Clinic offers an excellent salary and benefits package. Please apply online at www.mayoclinic.org and reference job posting #2103. Contact: Mayo Clinic Tina Cronk - Human Resources 200 1st Street SW, Rochester, MN 55905 Phone: 800-562-7984 From pruegg <@t> ihctech.net Thu Jan 27 16:09:40 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] endogenous peroxidase blues Message-ID: <200501272209.j0RM9apL014191@chip.viawest.net> Is there a sure fire way of iliminating endogenous peroxidase from granulecytes in acetone fixed cytospins for hrp IHC? Can I solve this problem without changing to a non-hrp detection enzyme? I am trying to stain T lymphocytes in cytospins of sputum. I do get cd3 positive lymphocyte looking cells, but agressive efforts has not completely blocked endogenous peroxidase in some of the larger cells with granules. Thanks for your help, Patsy From gcallis <@t> montana.edu Thu Jan 27 17:51:22 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] endogenous peroxidase blues In-Reply-To: <200501272209.j0RM9apL014191@chip.viawest.net> References: <200501272209.j0RM9apL014191@chip.viawest.net> Message-ID: <6.0.0.22.1.20050127165016.01b33078@gemini.msu.montana.edu> Glucose oxidase endogenous peroxidase and pseudoperoxidase block, I will send to you privately. It works on cells, frozen sections AND paraffin sections. At 03:09 PM 1/27/2005, you wrote: >Is there a sure fire way of iliminating endogenous peroxidase from >granulecytes in acetone fixed cytospins for hrp IHC? Can I solve this >problem without changing to a non-hrp detection enzyme? I am trying to >stain T lymphocytes in cytospins of sputum. I do get cd3 positive >lymphocyte looking cells, but agressive efforts has not completely blocked >endogenous peroxidase in some of the larger cells with granules. >Thanks for your help, >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 ctsblack <@t> capeheart.uct.ac.za Thu Jan 27 23:58:59 2005 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Thanks Message-ID: Thank you all for the methods you kindly sent me on Glycine HCl buffer. It is much appreciated. Thanx 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 JGordon <@t> cellmarque.com Fri Jan 28 00:41:11 2005 From: JGordon <@t> cellmarque.com (Jeff Gordon) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] endogenous peroxidase blues Message-ID: Patsy, we have a product that would probably help you out. It's called Peroxfree, and it is an endogenous peroxide block that can be used on any staining medium because it isn't hydrogen peroxide, so it doesn't lyse the cells like H2O2 would. Because of this feature, it is especially useful for cytosmears and cytospins. Check out our website for the product: http://www.cellmarque.com/products/reagents.php If you have any further questions, let us know. We would be happy to send you more information. 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 Patsy Ruegg Sent: Thursday, January 27, 2005 4:10 PM To: Histonet@Pathology.swmed.edu Subject: [Histonet] endogenous peroxidase blues Is there a sure fire way of iliminating endogenous peroxidase from granulecytes in acetone fixed cytospins for hrp IHC? Can I solve this problem without changing to a non-hrp detection enzyme? I am trying to stain T lymphocytes in cytospins of sputum. I do get cd3 positive lymphocyte looking cells, but agressive efforts has not completely blocked endogenous peroxidase in some of the larger cells with granules. Thanks for your help, Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Fri Jan 28 01:48:16 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] processing help[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3EEDB@bhrv-nt-11.bhrv.nwest.nhs.uk> I personally would suggest, logically its fixation. Have you changed anything there? Are you receiving them in proper formalin or water? Happened to me once. PS Dermatopathologist's are funny creatures, don't let them smell your fear. -----Original Message----- From: Dorothy.L.Webb@HealthPartners.Com [mailto:Dorothy.L.Webb@HealthPartners.Com] Sent: 27 January 2005 20:34 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] processing help[Scanned] I have a dermatopathologist who complained to me that his slides on the skins we short processed yesterday looked very eosinophilic and the collagen was separating out. Does anyone out there have any ideas what the problem could be?? We changed nothing in our processor from one day to the next nor any of the times. OUr routine tissues from today looked great! What could possibly be the problem when a pathologist comes across with this problem?? HELP!! And Thanks!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Martina.Urbanek <@t> uibk.ac.at Fri Jan 28 04:26:38 2005 From: Martina.Urbanek <@t> uibk.ac.at (Martina Urbanek) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] de Olmos staining Message-ID: <1106907998.41fa135ee5ed0@web-mail1.uibk.ac.at> Hello, Everyone, Does anyone have a detailed protocol for de Olmos amino-cupric-silver stain which works on paraffin sections (does it even work on paraffin?)? We work with mice brains which are fixed in 10% formalin, but not perfused and we are looking for a technique to stain degenerated neuronal cells. Your help is greatly appreciated. Martina Urbanek Ms. Martina Urbanek Forschungslabor der Klin.Abt. f?r Neonatologie neonatal neuroscience research laboratory Med. University Innsbruck Innrain 66, 4th floor A-6020 Innsbruck Tel. +43 (0)512 504 27755/27765 Fax: +43 (0)512 504 27766 Email: Martina.Urbanek@uibk.ac.at From vandries <@t> vub.ac.be Fri Jan 28 04:50:40 2005 From: vandries <@t> vub.ac.be (Veronique Andriessen) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] endogenous peroxidase blues In-Reply-To: <200501272209.j0RM9apL014191@chip.viawest.net> Message-ID: Hi Patsy, I have encountered somewhat of the same problem with endogenous peroxidase in acetone fixed frozen sections of acute pancreatitis, which is invaded with white blood cells. I tested several ways of blocking, home made and commercial including glucose oxidase. I never got it to work properly (believe me, I've tried!). What worked for me in the end was a new product by DAKO called "Dual Endogenous Enzyme Blocking Reagent" (S2003) (Thank you Mr. van der Loos!) It should work on cell preparations, frozen tissue and FFPE tissue (according to the manual). I only have tested it on frozen tissue. Just 10 minutes incubation at RT. This is just my honest opinion, I hope this can help you. Good luck Veronique Andriessen BAS Lab. Molecular Liver Cell Biology Free University Brussels (VUB), Belgium -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: donderdag 27 januari 2005 23:10 To: Histonet@Pathology.swmed.edu Subject: [Histonet] endogenous peroxidase blues Is there a sure fire way of iliminating endogenous peroxidase from granulecytes in acetone fixed cytospins for hrp IHC? Can I solve this problem without changing to a non-hrp detection enzyme? I am trying to stain T lymphocytes in cytospins of sputum. I do get cd3 positive lymphocyte looking cells, but agressive efforts has not completely blocked endogenous peroxidase in some of the larger cells with granules. Thanks for your help, Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Muskett <@t> RLC.NHS.UK Fri Jan 28 08:58:28 2005 From: David.Muskett <@t> RLC.NHS.UK (Muskett David) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Senior Biomedical Scientist vacancy in Liverpool UK[Scanned] Message-ID: <6AB79F0BA7C4914BA8DD8F5E0C21B9752E2897@AHEXMAIL01.xalderhey.com> Dear all Just a short note to advertise a vacancy within our department. ________________________________ ROYAL LIVERPOOL CHILDREN'S NHS TRUST, ALDER HEY (Working Towards Equal Opportunities) Post : Senior Biomedical Scientist (BMS2)- Paediatric Histopathology Hours: 37 Salary : ?21,287-26,937 ( discretionary points may be available to an exceptional candidate) The Royal Liverpool Children's NHS Trust is one of the largest paediatric hospitals in Western Europe. As a centre of excellence the trust treats more than 200 000 children a year from the North West , North Wales , Shropshire and the Isle of Man . Liverpool is a vibrant and lively city, its nightlife and culture is well known. The city is to be the European Capital of Culture in 2008 .This combined with modest house prices and easy access to the sea and countryside makes Liverpool the ideal location to live. The Histopathology Department has full CPA accreditation and is accredited for training with the IBMS . The department has recently moved to a purpose built laboratory and mortuary facility which won an award for 'Special Recognition for Service Design' at the 'Building Better Healthcare Awards 2003'. The department provides a full diagnostic paediatric histopathology service including immunocytochemistry and acetyl cholinesterase methods. The department provides a managed clinical network in fetal and perinatal pathology for the Merseyside and Cheshire Region. Applications are invited from experienced Registered Biomedical Scientists. You will be expected to possess a MSc. in Biomedical Sciences, and / or FIBMS with extensive experience of all aspects of histopathology. You will be expected to show evidence of continuing professional development. You will be responsible for the specialist techniques within the laboratory (immunocytochemistry, enzyme histochemistry, and fluorescent in-situ hybridisation) as well as participating in the routine work. You will take part in the organ retention work of the department and be expected to participate in the training of other staff. The department is committed to the development of its staff, who are encouraged to register for higher degrees and the IBMS Professional qualifications . Informal discussions are welcomed by David Muskett, Chief Biomedical Scientist, Histology on 0151 293 3656 e-mail david.muskett@rlc.nhs.uk The Royal Liverpool Children's NHS trust is committed to achieving equal opportunities in employment. The trust is committed to carefully screening all job applicants to ensure the safeguard of children. Application forms and job description are available from Recruit Direct on Telephone number 0151 529 8899. Please quote reference number Closing Date: Monday 21st February 2005 We operate a restricted smoking policy. From AJENKINS <@t> dental.ufl.edu Fri Jan 28 09:49:53 2005 From: AJENKINS <@t> dental.ufl.edu (Alan Jenkins) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] confirm i want to be added Message-ID: confirm i want to be added From pathrm35 <@t> adelphia.net Fri Jan 28 09:52:43 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] charge codes for toenails Message-ID: <3364338.1106927563731.JavaMail.root@web8.mail.adelphia.net> Fellow Techs, What charge codes are you using for nails? We do an H&E and a PAS for fungus. We also soften the nail with amm. hydroxide before cutting it. Is there a charge that can apply for the softening aspect of it? I know this is not a true decal but would the decal charge apply? Thanks, Ron From AJENKINS <@t> dental.ufl.edu Fri Jan 28 10:02:43 2005 From: AJENKINS <@t> dental.ufl.edu (Alan Jenkins) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Has anyone tried decalifying bone for frozen sections without fixing first? Message-ID: Has anyone tried decalifying bone for frozen sections without fixing first? From tom_c_nguyen <@t> yahoo.com Fri Jan 28 10:08:31 2005 From: tom_c_nguyen <@t> yahoo.com (Tom C. Nguyen) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Tissue Sectioning Message-ID: <20050128160831.23994.qmail@web51109.mail.yahoo.com> Thank you to everyone that replied to my previous inquiry regarding a device that can reliably cut tissue into 1mm slices. I'm a physician/surgeon (Stanford University) by trade and all this is very new to me. I've narrowed my choices to either the Vibratome or the EMS 5000 tissue slicer. Does anyone out there have personal hands on experience with these devices? If so, could you offer your thoughts or possibly give me some contact information. Thanks again. -tom http://scalpel.stanford.edu http://surgery.stanford.edu From vazquezr <@t> ohsu.edu Fri Jan 28 10:33:27 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Has anyone tried decalifying bone for frozen sections without fixing first? Message-ID: Alan, I do a 10% hydrochloric acid to decal for about 30min rinse, mount, cut and stain (which I have to baby the whole way) and I mainly do nasal bone. Robyn OHSU >>> "Alan Jenkins" 1/28/2005 8:02:43 AM >>> Has anyone tried decalcifying bone for frozen sections without fixing first? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Fri Jan 28 10:45:44 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Has anyone tried decalifying bone for frozen sections without fixing first? Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0012C99D7@UTHEVS3.mail.uthouston.edu> Alan It would be most helpful to us if you would please let us know precisely what you wish to finally accomplish with your frozen sections. It is possible to demineralize before fixation and retain the basic structure of the hard tissues so that you can demonstrate Haversian canals, Volkmann canals, canaliculi etc. but generally any soft tissue will be macerated or at least greatly distorted. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Alan Jenkins Sent: Friday, January 28, 2005 10:03 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Has anyone tried decalifying bone for frozen sectionswithout fixing first? Has anyone tried decalifying bone for frozen sections without fixing first? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Dixon.Leslie <@t> mayo.edu Fri Jan 28 10:50:16 2005 From: Dixon.Leslie <@t> mayo.edu (Dixon, Leslie E.) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Osteoblast stain with NBT-BCIP Message-ID: <115ED45FBA6D3D4B89E7F40A9EB5339D01826D6D@excsrv60.mayo.edu> Good morning, I want to stain osteoblasts using their natural phosphatase activity with NBT-BCIP. I needed to find out what most feel the optimum pH of the NBT-BCIP solution should be. I currently have liquid NBT-BCIP and will be diluting that if need be. Thanks in advance, Leslie From Jessica.Vaughn <@t> UCHSC.edu Fri Jan 28 11:19:03 2005 From: Jessica.Vaughn <@t> UCHSC.edu (Jessica.Vaughn@UCHSC.edu) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] B-gal staining of frozen mouse tissue Message-ID: <6294A7A909EDBF4FB5BD6FCDD46F0B4DCDE74C@hscex6> I am at wit's end when it comes to staining my frozen tissue of mouse lung (late stage embryo and newborn) for B-gal (w/ the enzyme, not Ab). Actually, I know that the staining works, the problem I am having is that whenever I go through the staining procedure, my tissue falls off of the slides. I have tried slides coated with gelatin, poly-l-lysine, and silane, and it never fails - the tissue falls off. I don't know if it is some sort of chemical reaction that is taking place with the staining solution or what it happening. I was just wondering if anyone else has had this problem and if anyone has any tips or tricks on how to overcome this issue. Please let me know if you do or if you have any protocols I could try that work for you. Thanks so much - I hope someone has some good advice for me before I lose my sanity! Jessica J. Vaughn Professional Research Assistant Dept. of Pediatrics University of Colorado Health Sciences Center From gcallis <@t> montana.edu Fri Jan 28 11:22:16 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Re: Has anyone tried decalifying bone for frozen sections without fixing first? In-Reply-To: References: Message-ID: <6.0.0.22.1.20050128100912.01b27ef0@gemini.msu.montana.edu> It has been done and is published. I have not done it. 1) Jonsson R et al. A demineralization procedure for immunohistopathological use: EDTA treatment preserves lymphoid cell surface antigens J Immunological Methods 88:109114,1986. This group had very good staining after a LONG EDTA decalcification on murine hindlegs (14 days) , a rather tedious protocol but their CD markers were well stained and tissue was preserved. After snap freezing EDTA decalcified bone, they cut frozen sections, and fixed them with acetone, did IHC for T and B lymphocytes, Ia antigens. 2) Kim L. Kusser and Troy D. Randall Simultaneous Detection of EGFP and Cell Surface Markers by Fluorescence Microscopy in Lymphoid Tissues J. Histochem. Cytochem., Jan 2003; 51: 5 - 14. Kusser also cites other publications in her article that may be helpful. At 09:02 AM 1/28/2005, you wrote: >Has anyone tried decalifying bone for frozen sections without fixing >first? >_______________________________________________ >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 Fri Jan 28 12:48:05 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] endogenous peroxidase blues In-Reply-To: Message-ID: <200501281848.j0SIm0pL025362@chip.viawest.net> Thank you all. You all are such a great resource, it saves me so much time to not have to reinvent the wheel everytime something troubling comes up. I am wondering if others have tried this Dual Endogenous Block from Dako for granulocytes? Patsy -----Original Message----- From: Veronique Andriessen [mailto:vandries@vub.ac.be] Sent: Friday, January 28, 2005 3:51 AM To: Histonet; Patsy Ruegg Subject: RE: [Histonet] endogenous peroxidase blues Hi Patsy, I have encountered somewhat of the same problem with endogenous peroxidase in acetone fixed frozen sections of acute pancreatitis, which is invaded with white blood cells. I tested several ways of blocking, home made and commercial including glucose oxidase. I never got it to work properly (believe me, I've tried!). What worked for me in the end was a new product by DAKO called "Dual Endogenous Enzyme Blocking Reagent" (S2003) (Thank you Mr. van der Loos!) It should work on cell preparations, frozen tissue and FFPE tissue (according to the manual). I only have tested it on frozen tissue. Just 10 minutes incubation at RT. This is just my honest opinion, I hope this can help you. Good luck Veronique Andriessen BAS Lab. Molecular Liver Cell Biology Free University Brussels (VUB), Belgium -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: donderdag 27 januari 2005 23:10 To: Histonet@Pathology.swmed.edu Subject: [Histonet] endogenous peroxidase blues Is there a sure fire way of iliminating endogenous peroxidase from granulecytes in acetone fixed cytospins for hrp IHC? Can I solve this problem without changing to a non-hrp detection enzyme? I am trying to stain T lymphocytes in cytospins of sputum. I do get cd3 positive lymphocyte looking cells, but agressive efforts has not completely blocked endogenous peroxidase in some of the larger cells with granules. Thanks for your help, Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From moss <@t> cut.net Fri Jan 28 15:40:43 2005 From: moss <@t> cut.net (MOSS) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] OT: Help with TEM work Message-ID: Greetings all. If there is anyone out there with an overload of TEM work, please contact me privately. We may be able to help. Connie McManus Mt Ogden Scientific Services 950 W Kershaw, Suite E Ogden UT 84401 tel: 866/933-6677 fax: 435/514-1781 moss@cut.net or conniemoss@relia.net From jefthompson <@t> salud.unm.edu Fri Jan 28 15:48:05 2005 From: jefthompson <@t> salud.unm.edu (Jeffrey Thompson) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Hello Histonet, Message-ID: Hello Histonet, I would like to stain 10 um rat sections for oligodendrocytes. The brains were fixed with PLP and then cryosectioned. Any suggestions for antibodies that people have had good resultes with (or those to avoid) would be greatly appreciated. Thanks, Jeff Thompson Jeffrey F. Thompson Associate Scientist University of New Mexico Health Sciences Center Department of Neurology BRF Room 136 915 Camino de Salud NE Albuquerque, NM 87131 USA jefthompson@salud.unm.edu Phone: (505) 272-8010 FAX: (505) 272-0607 From BRIAN.CHELACK <@t> usask.ca Fri Jan 28 17:31:31 2005 From: BRIAN.CHELACK <@t> usask.ca (Brian Chelack) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Re: endogenous peroxidase blocking in acetone fixed preps. Message-ID: <41FACB53.6ECB5B4C@sask.usask.ca> I have had success using the following treatment: 1% H2O2 0.1% Na Azide in PBS add to acetone fixed section/cell prep/smear for 10-12 minutes rinse 3X with PBS Carry on with blocking, primary antibody etc. regards, Brian Chelack From ynwang <@t> u.washington.edu Fri Jan 28 17:55:32 2005 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] cell characterization labels/stain Message-ID: I have a question about cell characterization. I know this isn't exactly your normal histology question but I've learnt so much from the people on this list in the past I was hoping someone would have a suggestion. We have started a primary cell culture from porcine anterior cruciate ligament (ACL). We have so far harvested the ACL, minced it and placed the pieces in culture to let the cells grow out of the pieces. After 5 days or so we have cell out growth from the pieces, however, we have cells with a variety of cell morphologies. I always thought we could generally tell cell type from the cell morphology however, it is proving to be rather difficult as we have a range of 'spindle-like' cells from very spindle-like to sort of spindle like but a bit more triangular. So, my question is: Does anyone know of a label/stain that can be used to characterize (ideally ACL) fibroblasts so that we can prove that our cells are ligament fibroblasts and not something else. I have done a search in some cell culture books, on the web and of past papers. I've even found images of what people think are ligament fibroblasts growing out of ligament pieces (ACL and MCL) however, I can't seem to find any information on characterizing these cells with something more reliable than saying 'these are fibroblasts because they have the characteristic spindle-like morphology'(am I not looking in the right places or with the correct key words?). The closest I've come was a paper on primary culture of urethra fibroblasts, which they identified as being fibroblasts because they were not smooth muscle cells (labeled for SM actin) or endothelial cells (labeled with an endothelial cell marker)! Any suggestions would be greatly appreciated. Yak-Nam Wang Senior Fellow Department of Bioengineering University of Washington Box 357962 Seattle, WA 98195 Tel.: (206)-221-5873 Fax.: (206)-221-5874 From jnocito <@t> satx.rr.com Sat Jan 29 08:42:49 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] charge codes for toenails References: <3364338.1106927563731.JavaMail.root@web8.mail.adelphia.net> Message-ID: <014201c50610$ce15dd70$ce22f318@yourxhtr8hvc4p> Ron, we have become the toenail capitol of South Texas remember "Joe the Toe"? We charge an 88304 for the nail itself and an 88312 for the PAS/Fungus. We use softening techniques also, but can't charge for them. Basically, nails are keratin and nor bone, therefore no decal. Of course, we always err on the side of caution because if we get audited by Medicare /Medicaid, we would be hard pressed to explain the decal charges. Joe Nocito BS, HT(ASCP)QIHC a.k.a Joe the Toe Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: To: Sent: Friday, January 28, 2005 9:52 AM Subject: [Histonet] charge codes for toenails > Fellow Techs, > What charge codes are you using for nails? We do an H&E and a PAS for > fungus. We also soften the nail with amm. hydroxide before cutting it. Is > there a charge that can apply for the softening aspect of it? I know this > is not a true decal but would the decal charge apply? > Thanks, > Ron > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From subratab <@t> bdonline.com Sat Jan 29 12:18:56 2005 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] HRP-conjugated secondary and biotinylated-secondary Message-ID: <200501291823.j0TINDYb016016@mailout.proshikanet.com> Hi, Is it a common practice to use HRP-conjugated secondary antibody instead of biotinylated secondary antibody for immunohistochemistry (with DAB developing system)? I am confused about the advantage/ disadvantage of using HRP-conjugated secondary antibody for immunos. Subrata Biswas UNICAMP, SP, Brazil. From pruegg <@t> ihctech.net Sun Jan 30 14:39:42 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] HRP-conjugated secondary and biotinylated-secondary In-Reply-To: <200501291823.j0TINDYb016016@mailout.proshikanet.com> Message-ID: <200501302042.j0UKfsZ0046090@pro12.abac.com> Subrata, An advantage to using hrp conjugated secondary antibody instead of biotinylated is that you can avoid the possible interference of endogenous biotin which for some tissues may be difficult or impossible to get rid of in my experience. A disadvantage is that the indirect, two step hrp conjugated method is not as sensitive as the ABC method (you build on to the molecule by having more binding sites with the ABC method than with the hrp conjugated method. My choice for detection when possible is to use a labeled polymer which is conjugated to hrp. The sensitivity is as great or greater as the ABC method without the endogenous biotin problem. Patsy Ruegg -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Subratab Sent: Saturday, January 29, 2005 11:19 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HRP-conjugated secondary and biotinylated-secondary Hi, Is it a common practice to use HRP-conjugated secondary antibody instead of biotinylated secondary antibody for immunohistochemistry (with DAB developing system)? I am confused about the advantage/ disadvantage of using HRP-conjugated secondary antibody for immunos. Subrata Biswas UNICAMP, SP, Brazil. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jason.PALMER <@t> svhm.org.au Sun Jan 30 23:10:34 2005 From: Jason.PALMER <@t> svhm.org.au (PALMER Jason (SVHM)) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] queries re freezing PFA, and storing tissue after fixation Message-ID: Hi histonetters. I know there has been quite a lot written on histonet about how formaldehyde fixatives work, and comparisons between PFA vs buffered formalin solutions for fixation etc, and I have read many of these, but there are still a couple of issues relevant to my lab that I'm not quite sure about. I work in a research setting and the tissues we process are mostly mouse or rat, and quite small, I think - say 3 x 6 mm on average, or something like that. We do lots of immunohistochemistry, and normally fix our samples in 4% PFA / PBS for 24 hrs at 4 degrees C. There are a couple of practices in our lab that worry me a little, and I'm not sure who instigated them and nobody can give me good explanations why they are acceptable. One is the use of PFA that has been frozen. It is made up in the usual way, aliqoutted and put into the -20 c freezer on the same day, stored there, and thawed out on the day (or day before) of use. Making PFA in one biggish lot and freezing aliquots makes sense in terms of saving time and effort, but is the PFA going to remain "stable" and not start to repolymerise over what could be weeks, or even months, in the -20 c freezer?? Personally, I'm not even convinced that freshly made and used PFA is better than 10% buffered formalin for our immuno work, on the whole, though that's another issue... Secondly, it is often the case here that after fixation, tissues are stored in PBS (in the 4 degree C fridge) for what may be weeks rather than days before processing. I personally haven't notices any difference in the immunostaining seen with these samples, compared with samples processed immediately or within days, for a number of antigens, but I still feel that storage for weeks in PBS is not a good idea. From what I've read, formaldehyde fixation for the time we use (24 hrs or sometimes even a little less) is reversible over time, in PBS especially. I guess an alternative is to store in 70% ethanol, but again I can't quite help wondering if this might not be conducive to good and consistent immunostaining. Any thoughts much appreciated, 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 -------------- 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 c.m.vanderloos <@t> amc.uva.nl Mon Jan 31 01:50:22 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] RE: endogenous peroxidase blues Message-ID: <8a45c28abdde.8abdde8a45c2@amc.uva.nl> Hi all, Before everybody is trying to solve their problem of endogenous peroxidase activity with Dual Enzyme Blocking Reagent as Veronique wrote us, I would like to make a comment. This stuff works great indeed in many cases, that's true. But I also found some antigens that didn't survive this treatement at all. So if you are going to test Dual Enzyme Blocking Reagent, keep this in mind and be sure to have the right controls involved. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From "Veronique Andriessen" Date Fri, 28 Jan 2005 11:50:40 +0100 To "Histonet" , "Patsy Ruegg" Subject RE: [Histonet] endogenous peroxidase blues Hi Patsy, I have encountered somewhat of the same problem with endogenous peroxidase in acetone fixed frozen sections of acute pancreatitis, which is invaded with white blood cells. I tested several ways of blocking, home made and commercial including glucose oxidase. I never got it to work properly (believe me, I've tried!). What worked for me in the end was a new product by DAKO called "Dual Endogenous Enzyme Blocking Reagent" (S2003) (Thank you Mr. van der Loos!) It should work on cell preparations, frozen tissue and FFPE tissue (according to the manual). I only have tested it on frozen tissue. Just 10 minutes incubation at RT. This is just my honest opinion, I hope this can help you. Good luck Veronique Andriessen BAS Lab. Molecular Liver Cell Biology Free University Brussels (VUB), Belgium -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu[ References 1. mailto:c.m.vanderloos@amc.uva.nl From c.m.vanderloos <@t> amc.uva.nl Mon Jan 31 01:55:53 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Re: endogenous peroxidase blocking in acetone fixed preps Message-ID: <14ed81d14eb1f3.14eb1f314ed81d@amc.uva.nl> Dear Brian, The solution as you suggested works fine indeed for abolishing peroxidase activity in erythrocytes. However, it kills the endogenous peroxidase activity in neutrophils just marginally. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From Brian Chelack Date Fri, 28 Jan 2005 17:31:31 -0600 To Histonet@Pathology.swmed.edu Subject [Histonet] Re: endogenous peroxidase blocking in acetone fixed preps. I have had success using the following treatment: 1% H2O2 0.1% Na Azide in PBS add to acetone fixed section/cell prep/smear for 10-12 minutes rinse 3X with PBS Carry on with blocking, primary antibody etc. regards, Brian Chelack References 1. mailto:c.m.vanderloos@amc.uva.nl From c.m.vanderloos <@t> amc.uva.nl Mon Jan 31 02:23:42 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] RE: HRP-conjugated secondary and biotinylated-secondary Message-ID: Hi, The existence of both biotinylated- and HRP-conjuated secondary antibodies reflects the development of IHC techniques into more staining efficiency/sensitivity. In the old days, a 2-step staining method using a HRP-conjugated secondary anti-mouse or anti-rabbit was common practice. Mid 80-ies the 3-step staining method using a biotinylated 2nd step reagent and streptavidin/HRP conjugate or streptavidin-biotinylated HRP complex became very popular because of its superior staining efficiency/sensitivity. The disadvantage here is the occurrence of endogenous biotin in many tissues. Endogenous biotin is not present in untreated formalin-fixed and paraffin-embedded tissue sections, but will be retrieved especially when using Tris-EDTA pH8-9. Only recently we have gone to a 2-step staining procedure again, now based on both secondary antibody and HRP attached to a polymer backbone. The efficiency/sensitivity of these polymers is comparable with the 3-step streptavidin/HRP procedure, but avoids the problem of endogenous biotin. I hope this small historical overview helps. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From Subratab Date Sun, 30 Jan 2005 0:18:56 +0600 To Subject [Histonet] HRP-conjugated secondary and biotinylated-secondary Hi, Is it a common practice to use HRP-conjugated secondary antibody instead of biotinylated secondary antibody for immunohistochemistry (with DAB developing system)? I am confused about the advantage/ disadvantage of using HRP-conjugated secondary antibody for immunos. Subrata Biswas UNICAMP, SP, Brazil. References 1. mailto:c.m.vanderloos@amc.uva.nl From colins <@t> thor.uk.com Mon Jan 31 03:34:31 2005 From: colins <@t> thor.uk.com (Colin Smith) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Manual fixation/processing of reconstructedskincultures grown on polycarbonate filters Message-ID: <3E985CD1D7CA89499D1902BC3202AF3CC1A762@uk-res-srv-03.thorspecialities.com> Liz, Thanks for your help. I would be interested in a reprint of the article you are writng for Histologic. Best Regards, Colin -----Original Message----- From: Elizabeth Chlipala [mailto:liz@premierlab.com] Sent: 26 January 2005 16:36 To: Colin Smith; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Manual fixation/processing of reconstructedskincultures grown on polycarbonate filters Colin We have worked quite a bit with the epioral and epigingival cultures from Mattek. We have stained with H&E, Ki-67 and Cleaved caspase 3. We processed these specimens on a tissue processor, but you will be able to do this by hand. I'm not sure of the size of your culture wells, but we used an 8mm biopsy punch to remove the constructs from the wells, processed the construct whole between two biopsy pads and bisected with a razor blade prior to embedding on end. As far as fixation 10% NBF will work. The wells were labeled on the side and the entire well was placed in a 50 ml conical tube with fixative. After adequate fixation the specimens were grossed, processed and embedded into paraffin. We processed during the day (not overnight) at 15 minutes per station. We sectioned at 4 microns and stained with both H&E and various Immunohistochemical stains, I can send you images if you are interested. If you need more info, give me a call or e-mail me back. Also we are writing an article that will be in the spring edition of Histologic that basically covers the methods (both processing and analysis) we used to assess proliferation in these in vitro tissue models. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Colin Smith Sent: Wednesday, January 26, 2005 8:07 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Manual fixation/processing of reconstructed skincultures grown on polycarbonate filters Can anyone help? I am currently setting up a basic histology system for analysing reconstructed tissues grown in culture on polycarbonate filters, similar to Mattek's Epiderm. We currently have begged borrowed or stolen, equipment-wise: two water baths (ambient - 100C), a circulating warm air oven, a rocking microtome and cold blocks, along with glass staining jars/racks, forceps etc. At the moment, equipment such as a wax dispenser, purpose built cooling plates etc are out of the question until I can show that we can show that the model we are growing is validated, which will include immunohistochemistry of cytokeratin and cell adhesion markers etc. Thus we are in the catch 22 situation of having to prove something without the equipment needed to do the job, in order to justify buying the equipment!! But thats accountants for you, go figure. Anyway, can anyone give any useful pointers/protocols for fixing/processing/embedding/sectioning such tissues using such primative equipment Any help, no matter how trivial will be greatfully received. Many Thanks in Advance, Colin Smith BSc CBiol MIBiol Eurotox Registered Toxicologist In Vitro Toxicologist Thor Specialities UK Ltd Wincham Avenue, Northwich CW9 6GB Tel: +44 1606 818873 Fax: +44 1606 818801 colins@thor.uk.com The information contained in this e-mail may be confidential and may also be legally privileged. It is intended solely for the addressee(s). If you are not the intended recipient then please delete forthwith. Views expressed in this email are views of the individual and are not necessarily the views of this company. ************************************************************************ ************************* e-mail scanned for Viruses by Thor Specialities (UK) Limited using Gfi Mail Security ************************************************************************ ************************* _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Mon Jan 31 09:25:40 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] RE: endogenous peroxidase blues In-Reply-To: <8a45c28abdde.8abdde8a45c2@amc.uva.nl> Message-ID: <200501311527.j0VFRq0G018003@pro12.abac.com> Chris can you share with us which antigens didn't survive this enzyme block in your hands. Thanks, Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of C.M. van der Loos Sent: Monday, January 31, 2005 12:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: endogenous peroxidase blues Hi all, Before everybody is trying to solve their problem of endogenous peroxidase activity with Dual Enzyme Blocking Reagent as Veronique wrote us, I would like to make a comment. This stuff works great indeed in many cases, that's true. But I also found some antigens that didn't survive this treatement at all. So if you are going to test Dual Enzyme Blocking Reagent, keep this in mind and be sure to have the right controls involved. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From "Veronique Andriessen" Date Fri, 28 Jan 2005 11:50:40 +0100 To "Histonet" , "Patsy Ruegg" Subject RE: [Histonet] endogenous peroxidase blues Hi Patsy, I have encountered somewhat of the same problem with endogenous peroxidase in acetone fixed frozen sections of acute pancreatitis, which is invaded with white blood cells. I tested several ways of blocking, home made and commercial including glucose oxidase. I never got it to work properly (believe me, I've tried!). What worked for me in the end was a new product by DAKO called "Dual Endogenous Enzyme Blocking Reagent" (S2003) (Thank you Mr. van der Loos!) It should work on cell preparations, frozen tissue and FFPE tissue (according to the manual). I only have tested it on frozen tissue. Just 10 minutes incubation at RT. This is just my honest opinion, I hope this can help you. Good luck Veronique Andriessen BAS Lab. Molecular Liver Cell Biology Free University Brussels (VUB), Belgium -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu[ References 1. mailto:c.m.vanderloos@amc.uva.nl _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kspencer <@t> utmem.edu Mon Jan 31 09:43:12 2005 From: kspencer <@t> utmem.edu (Kathleen Spencer) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] queries re freezing PFA, and storing tissue after fixation In-Reply-To: References: Message-ID: We store rat brains in 20% sucrose in the fridge until we take them out for cryosectioing. Our animals are perfused with very fresh 4% PFA only. Kathleen On Jan 30, 2005, at 11:10 PM, PALMER Jason (SVHM) wrote: > > Hi histonetters. > > I know there has been quite a lot written on histonet about how > formaldehyde fixatives work, and comparisons between PFA vs buffered > formalin solutions for fixation etc, and I have read many of these, > but there are still a couple of issues relevant to my lab that I'm not > quite sure about. > > I work in a research setting and the tissues we process are mostly > mouse or rat, and quite small, I think - say 3 x 6 mm on average, or > something like that. We do lots of immunohistochemistry, and normally > fix our samples in 4% PFA / PBS for 24 hrs at 4 degrees C. There are a > couple of practices in our lab that worry me a little, and I'm not > sure who instigated them and nobody can give me good explanations why > they are acceptable. One is the use of PFA that has been frozen. It is > made up in the usual way, aliqoutted and put into the -20 c freezer on > the same day, stored there, and thawed out on the day (or day before) > of use. Making PFA in one biggish lot and freezing aliquots makes > sense in terms of saving time and effort, but is the PFA going to > remain "stable" and not start to repolymerise over what could be > weeks, or even months, in the -20 c freezer?? Personally, I'm not even > convinced that freshly made and used PFA is better than 10% buffered > formalin for our immuno work, on the whole, though that's another > issue... > > Secondly, it is often the case here that after fixation, tissues are > stored in PBS (in the 4 degree C fridge) for what may be weeks rather > than days before processing. I personally haven't notices any > difference in the immunostaining seen with these samples, compared > with samples processed immediately or within days, for a number of > antigens, but I still feel that storage for weeks in PBS is not a good > idea. From what I've read, formaldehyde fixation for the time we use > (24 hrs or sometimes even a little less) is reversible over time, in > PBS especially. I guess an alternative is to store in 70% ethanol, but > again I can't quite help wondering if this might not be conducive to > good and consistent immunostaining. > > Any thoughts much appreciated, > > 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 > > _____________________________________________ > __ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tessajmurray <@t> hotmail.com Mon Jan 31 10:00:33 2005 From: tessajmurray <@t> hotmail.com (Tessa Murray) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] IHC in resin embedded sections Message-ID: Dear Histonetters, I am looking for some advice about the feasibility of doing IHC on tissues embedded for EM work. I need to section very small tissues in a specific orientation which is difficult to determine when embedding in paraffin. I was wondering if we could borrow the EM technique where (as I understand) the tissue is embedded in resin and then temporarily fixed to a "peg" for sectioning - thus allowing us to cut some sections, check the orientation and then rotate the block accordingly until we get the orientation exactly right. Obviously I am concerned about how well antibodies are going to recognize the tissue after processing, how are the sections de-resined, is there a specific antigen retreival step for resin tissues etc. Any help appreciated - perhaps there is a way to adjust sectioning angles in FFPE tissues that I've not thought of? Frozen tissues are not an option as we want to preserve morphology as much as possible. Cheers guys. Tessa J Murray PhD Tufts University School of Medicine From am458 <@t> cornell.edu Mon Jan 31 10:38:24 2005 From: am458 <@t> cornell.edu (Amanda MacFarlane) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] aqueous mounting medium Message-ID: <1864.132.236.104.213.1107189504.squirrel@132.236.104.213> I have stained whole tissues for beta-galactosidase (X-gal) and am now cryosectioning the tissues. I have been fixing the sections in 4% PFA overnight at 4 degrees Celsius and cover slipping with 70% glycerol. Coverslipping with glycerol is a time-consuming pain in the butt so I've checked out glycerol vinyl alcohol aqueous (GVA) mounting solution (Zymed) and Supermount aqueous mounting medium (Biogenex) as alternatives. It appears that these products have not been tested with beta-gal staining, so I'm wondering if people use these products and/or what people recommend. Thanks. Amanda MacFarlane, PhD Division of Nutritional Sciences Cornell University From Gina.Sennello <@t> osip.com Mon Jan 31 10:46:52 2005 From: Gina.Sennello <@t> osip.com (Sennello, Gina) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] FW: total lobectomy Message-ID: > Hello everyone, > > I work in research was wondering if hospitals have protocols for the > number of blocks that are taken for total lung lobectomy cases. Is the > number of blocks determined by the stage of the cancer at the time of > diagnosis? > > Thanks in advance , > > Gina > > Gina Sennello > Senior Associate Scientist > Histotechnologist > > OSIP > 2860 Wilderness Place > Boulder, CO > 80301 > > phone 303-546-7739 > fax 303-444-0672 > From georgecole <@t> ev1.net Mon Jan 31 10:51:54 2005 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] Fixed VS unfixed tissues for immunofluorescence and antibody work Message-ID: <000001c507b5$2f428210$014dbad0@hppav> My histological career began in June 1962. When I was assigned immuno studies, the thinking of the day was that fixation blocked/ruined those reactions as well as most antibody preps. SO it went until about 1980 or so, when muscle and nerve biopsy work got so heavy, those immuno and antibody needs were met in the main lab. Somewhere in there, I got word that immuno and antibody studies were being done on fixed tissues. This was startling news to me---a reversal of an Aboslute. And I tell you, such is the power the question-generating innards of my head, I would have compared with-and-without-fixative preps of any immuno or antibody studies that I might have been assigned. But I was hard enough pressed to deal with my work load---I always picture biopsy patients , waiting anxiously for the results of their tissues. So the question of fixed-unfixed immunos and antibody work didn't really revive until I retired and became a regular visitor to the Histonet. I know that I am a born fuss budget and I would NEVER opt for ANY choice of diagnostic procedure without rating it with all candidate procedures for that specific study. So far, I have had no comforting word from the world-wide Histology community that EVERY immuno and antibody technique now being discussed on the Histonet had been cleared for action by initial comparisons of results with and without fixation. All I have seen in mentioning this picky-picky compulsion of mine, is the flat statement is THIS IS HOW WE DO IT!! The work being done in any lab is almost never reported with credentials of the procedures used---that that particular procedure was being done, because it had won a contest of effectiveness over other procedures. Furthermore, just beneath the surface, is the feeling that the "This is the way I do it. Period!!" attitude was lurking. Now, this aging retiree wishes to avoid conflict and arousing tempers in you younguns. Age does not cater to warfare. But I would treasure any reports from you active histotechs about the credentials of choice from among candidate procedures--- the winners of effectiveness resulting from comparisons of techniques that are rivals to your chosen ones. Such reports would receive an honored place, hanging on the wall here over my computer. I would honor them---and you would add peace to the Age-Naps of this old histotech!!. georgecole@ev1.net From gcallis <@t> montana.edu Mon Jan 31 11:16:07 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] aqueous mounting medium In-Reply-To: <1864.132.236.104.213.1107189504.squirrel@132.236.104.213> References: <1864.132.236.104.213.1107189504.squirrel@132.236.104.213> Message-ID: <6.0.0.22.1.20050131101120.01b52760@gemini.msu.montana.edu> Amanda, I have used the equivalent to Supermount, only it was the Biomeda Crystal mount. We did the liquid coverslip, generously over section, to let it set up then mounted a permanent coverslip over that. Worked great. Instructions tell you how to let this liquid coverslip dry before putting a permanent coverslip over the top. In general, we air dry our beta gal stained whole tissues, then counterstain with inhouse Nuclear fast red, rinse well with water, air dry and mount a coverslip with a permanent mounting media. No problems. We hate messy mounting media that cause coverslip glissading (slip sliding around), and poor storage of goo that never sets up. At 09:38 AM 1/31/2005, you wrote: >I have stained whole tissues for beta-galactosidase (X-gal) and am now >cryosectioning the tissues. I have been fixing the sections in 4% PFA >overnight at 4 degrees Celsius and cover slipping with 70% glycerol. >Coverslipping with glycerol is a time-consuming pain in the butt so I've >checked out glycerol vinyl alcohol aqueous (GVA) mounting solution (Zymed) >and Supermount aqueous mounting medium (Biogenex) as alternatives. It >appears that these products have not been tested with beta-gal staining, >so I'm wondering if people use these products and/or what people >recommend. Thanks. > > >Amanda MacFarlane, PhD >Division of Nutritional Sciences >Cornell University > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From ynwang <@t> u.washington.edu Mon Jan 31 15:50:38 2005 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] cell characterization labels/stain In-Reply-To: <0c5c01c505a6$196f4600$0200a8c0@desktop> References: <0c5c01c505a6$196f4600$0200a8c0@desktop> Message-ID: Diane, Thanks for your suggestion! I had a look at the website and spoke to their US distributor, unfortunately, no-one has tested it in porcine tissue. I still want to try to find out the epitopes to see if there is a possiblity of cross-reactivity. If I make any progress or I get any other responses I will definately let you know. Thanks again for the suggestion. Yak-Nam > Yak-- > > Couldn't you just stain with an antibody that stains Fibroblasts? > > of antibodies for staining Fibroblasts in human, mouse and rat tissues... ... Antibodies to human Fibroblasts. ... > www.acris-antibodies.com/focuson/focuson-new-08.html - 57k - Cached - Similar pages > > I'd be interested in hearing what responses you get. I'm working with endothelium cells. > > Diane > Senior Research Assistant > BioMedical Engineering, OGI, OHSU. > 503-784-6444 > ----- Original Message ----- > From: Y. Wang > To: Histonet@lists.utsouthwestern.edu > Sent: Friday, January 28, 2005 3:55 PM > Subject: [Histonet] cell characterization labels/stain > > > I have a question about cell characterization. I know this isn't exactly > your normal histology question but I've learnt so much from the people on > this list in the past I was hoping someone would have a suggestion. > > We have started a primary cell culture from porcine anterior cruciate > ligament (ACL). We have so far harvested the ACL, minced it and placed the > pieces in culture to let the cells grow out of the pieces. After 5 days or > so we have cell out growth from the pieces, however, we have cells with a > variety of cell morphologies. I always thought we could generally tell > cell type from the cell morphology however, it is proving to be rather > difficult as we have a range of 'spindle-like' cells from very > spindle-like to sort of spindle like but a bit more triangular. > > So, my question is: Does anyone know of a label/stain that can be used to > characterize (ideally ACL) fibroblasts so that we can prove that our cells > are ligament fibroblasts and not something else. > > I have done a search in some cell culture books, on the web and of past > papers. I've even found images of what people think are ligament > fibroblasts growing out of ligament pieces (ACL and MCL) however, I can't > seem to find any information on characterizing these cells with something > more reliable than saying 'these are fibroblasts because they have the > characteristic spindle-like morphology'(am I not looking in the right > places or with the correct key words?). The closest I've come was a paper > on primary culture of urethra fibroblasts, which they identified as being > fibroblasts because they were not smooth muscle cells (labeled for SM > actin) or endothelial cells (labeled with an endothelial cell marker)! > > Any suggestions would be greatly appreciated. > Yak-Nam Wang > > Senior Fellow > Department of Bioengineering > University of Washington > Box 357962 > Seattle, WA 98195 > > Tel.: (206)-221-5873 > Fax.: (206)-221-5874 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Mon Jan 31 19:10:11 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] IHC in resin embedded sections In-Reply-To: References: Message-ID: <41FED6F3.6060709@umdnj.edu> Your success will depend on the antigen in question and how it reacts to fixation and embedding. I would try fixing in buffered formalin or paraformaldehyde and embedding in Araldite or an Epon substitute, avoid Spurr's resin. There are some resins that are quite antigen friendly, Lowicryl for one, but they are a bit harder to use. You might be able to use some glutaraldehyde in the fixative but I would first find a method that works, then try for better morphology with stronger fixation. You might also try Vibratome sections of 40-100 microns before embedding. Then do IHC on those before embedding or just use those sections to check orientation of the tissue. Frozen sections of fixed material should provide very good morphology if freezing is done very quickly after proper cryoprotection. Geoff Tessa Murray wrote: > > Dear Histonetters, > > I am looking for some advice about the feasibility of doing IHC on > tissues embedded for EM work. I need to section very small tissues in > a specific orientation which is difficult to determine when embedding > in paraffin. I was wondering if we could borrow the EM technique > where (as I understand) the tissue is embedded in resin and then > temporarily fixed to a "peg" for sectioning - thus allowing us to cut > some sections, check the orientation and then rotate the block > accordingly until we get the orientation exactly right. Obviously I > am concerned about how well antibodies are going to recognize the > tissue after processing, how are the sections de-resined, is there a > specific antigen retreival step for resin tissues etc. Any help > appreciated - perhaps there is a way to adjust sectioning angles in > FFPE tissues that I've not thought of? Frozen tissues are not an > option as we want to preserve morphology as much as possible. Cheers > guys. > > Tessa J Murray PhD > Tufts University School of Medicine > _______________________________________________ > 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 gcallis <@t> montana.edu Mon Jan 31 17:34:17 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] More on Re: IHC in resin embedded sections In-Reply-To: <41FED6F3.6060709@umdnj.edu> References: <41FED6F3.6060709@umdnj.edu> Message-ID: <6.0.0.22.1.20050131162315.01b3bbe0@gemini.msu.montana.edu> There is a way to remove resin, but it is not very friendly at times to the tissue, called sodium ethoxide. Recipe for this can be found in Histonet archives, www.histosearch.org as it was discussed not too long ago. You can search for this info IF you think you need it. Also, antigen retrieval methods have been done on tissues embedded in resins for EM purposes. Shi and Taylors book, Antigen Retrieval Techniques has a whole chapter devoted to this. The book can be purchased from Eaton Publishing, and is worth having around a lab for other retrieval technics, hints, etc. I have seen retrieval methods published for EM resins, but you would have to hit PUBMED to find them, they can be found in EM type journals - it is something we do not do, but cruising journals can be fun. An aside - have heard Spurrs is being discontinued as one of the ingredients is not being made anymore. At 06:10 PM 1/31/2005, you wrote: > Your success will depend on the antigen in question and how it reacts > to fixation and embedding. >I would try fixing in buffered formalin or paraformaldehyde and embedding >in Araldite or an Epon substitute, avoid Spurr's resin. There are some >resins that are quite antigen friendly, Lowicryl for one, but they are a >bit harder to use. You might be able to use some glutaraldehyde in the >fixative but I would first find a method that works, then try for better >morphology with stronger fixation. > You might also try Vibratome sections of 40-100 microns before > embedding. Then do IHC on those before embedding or just use those > sections to check orientation of the tissue. > Frozen sections of fixed material should provide very good morphology > if freezing is done very quickly after proper cryoprotection. > >Geoff > >Tessa Murray wrote: > >> >> Dear Histonetters, >> >> I am looking for some advice about the feasibility of doing IHC on >> tissues embedded for EM work. I need to section very small tissues in >> a specific orientation which is difficult to determine when embedding >> in paraffin. I was wondering if we could borrow the EM technique >> where (as I understand) the tissue is embedded in resin and then >> temporarily fixed to a "peg" for sectioning - thus allowing us to cut >> some sections, check the orientation and then rotate the block >> accordingly until we get the orientation exactly right. Obviously I >> am concerned about how well antibodies are going to recognize the >> tissue after processing, how are the sections de-resined, is there a >> specific antigen retreival step for resin tissues etc. Any help >> appreciated - perhaps there is a way to adjust sectioning angles in >> FFPE tissues that I've not thought of? Frozen tissues are not an >> option as we want to preserve morphology as much as possible. Cheers >> guys. >> >> Tessa J Murray PhD >> Tufts University School of Medicine >>_______________________________________________ >>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 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 jfish <@t> gladstone.ucsf.edu Mon Jan 31 17:50:57 2005 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] More on Re: IHC in resin embedded sections In-Reply-To: <6.0.0.22.1.20050131162315.01b3bbe0@gemini.msu.montana.edu> References: <41FED6F3.6060709@umdnj.edu> <6.0.0.22.1.20050131162315.01b3bbe0@gemini.msu.montana.edu> Message-ID: Tessa, You can also use Unicryl. It is a resin that can be polymerized at low temperatures, e.g. 4 degrees under U.V. lamp. The resin does not need to be etched because when sectioned it is cleaved, exposing antigen sights. It is very easy to use, too. Take care, Jo Dee At 4:34 PM -0700 1/31/05, Gayle Callis wrote: >There is a way to remove resin, but it is not very friendly at times >to the tissue, called sodium ethoxide. Recipe for this can be found >in Histonet archives, www.histosearch.org as it was discussed not >too long ago. You can search for this info IF you think you need it. > >Also, antigen retrieval methods have been done on tissues embedded >in resins for EM purposes. Shi and Taylors book, Antigen >Retrieval Techniques has a whole chapter devoted to this. The book >can be purchased from Eaton Publishing, and is worth having around a >lab for other retrieval technics, hints, etc. I have seen >retrieval methods published for EM resins, but you would have to hit >PUBMED to find them, they can be found in EM type journals - it is >something we do not do, but cruising journals can be fun. > >An aside - have heard Spurrs is being discontinued as one of the >ingredients is not being made anymore. > > > >At 06:10 PM 1/31/2005, you wrote: >> Your success will depend on the antigen in question and how it >>reacts to fixation and embedding. >>I would try fixing in buffered formalin or paraformaldehyde and >>embedding in Araldite or an Epon substitute, avoid Spurr's resin. >>There are some resins that are quite antigen friendly, Lowicryl for >>one, but they are a bit harder to use. You might be able to use >>some glutaraldehyde in the fixative but I would first find a method >>that works, then try for better morphology with stronger fixation. >> You might also try Vibratome sections of 40-100 microns before >>embedding. Then do IHC on those before embedding or just use those >>sections to check orientation of the tissue. >> Frozen sections of fixed material should provide very good >>morphology if freezing is done very quickly after proper >>cryoprotection. >> >>Geoff >> >>Tessa Murray wrote: >> >>> >>> Dear Histonetters, >>> >>> I am looking for some advice about the feasibility of doing IHC on >>> tissues embedded for EM work. I need to section very small tissues in >>> a specific orientation which is difficult to determine when embedding >>> in paraffin. I was wondering if we could borrow the EM technique >>> where (as I understand) the tissue is embedded in resin and then >>> temporarily fixed to a "peg" for sectioning - thus allowing us to cut >>> some sections, check the orientation and then rotate the block >>> accordingly until we get the orientation exactly right. Obviously I >>> am concerned about how well antibodies are going to recognize the >>> tissue after processing, how are the sections de-resined, is there a >>> specific antigen retreival step for resin tissues etc. Any help >>> appreciated - perhaps there is a way to adjust sectioning angles in >>> FFPE tissues that I've not thought of? Frozen tissues are not an >>> option as we want to preserve morphology as much as possible. Cheers >>> guys. >>> >>> Tessa J Murray PhD >>> Tufts University School of Medicine >>>_______________________________________________ >>>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 > >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 ********************************************************************** ********** Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease Telephone: (415) 734-5766 Fax: (415) 355-0230 E-mail: jfish@gladstone.ucsf.edu Mailing address: The J. David Gladstone Institutes 1650 Owens Street San Francisco, CA 94158 From histomjans <@t> yahoo.com Mon Jan 31 19:40:03 2005 From: histomjans <@t> yahoo.com (Melissa Jans) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] yeast and bacteria on slides Message-ID: <20050201014003.84482.qmail@web54705.mail.yahoo.com> I need some help!! We have been experiencing some yeast and bacteria showing up on some of our special stains. It started about 2 weeks ago and seems to be sporadic (one day...no problems, the next day, several stains are affected). The yeast and bacteria (rods) are clumped together all over the slide, on the tissue sections, around the sections and on the slide in areas where there is no tissue. The stains we have seen the contamination are the Gram, PAS/D, PAS and GMS. All of these stains are automated except the Gram, so I do not think the special stainer is the problem. Here is what I have eliminated as problem areas: -Processor solutions and paraffin (we cut and stained a block from another institute and had contamination) -Waterbaths, slide drying boards, water containers (used to fill waterbathes), stainer containers, ice pans (all of the mentioned have been thoroughly bleached) -Tap water and DI water (all water supplies have been tested by sending a sample to micro...there was no bacteria or yeast detected) -Stay-on used as a slide adhesive (smeared some on a slide and stained it) -Bad batch of slides (we have had contamination on two different slide sources (vendors)) Does anybody have any ideas? What am I missing? Any suggestions would be much appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From failm <@t> musc.edu Mon Jan 31 19:51:12 2005 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Sep 16 15:24:33 2005 Subject: [Histonet] yeast and bacteria on slides Message-ID: Do you have tubing on your faucets? >>> Melissa Jans 01/31/05 20:41 PM >>> I need some help!! We have been experiencing some yeast and bacteria showing up on some of our special stains. It started about 2 weeks ago and seems to be sporadic (one day...no problems, the next day, several stains are affected). The yeast and bacteria (rods) are clumped together all over the slide, on the tissue sections, around the sections and on the slide in areas where there is no tissue. The stains we have seen the contamination are the Gram, PAS/D, PAS and GMS. All of these stains are automated except the Gram, so I do not think the special stainer is the problem. Here is what I have eliminated as problem areas: -Processor solutions and paraffin (we cut and stained a block from another institute and had contamination) -Waterbaths, slide drying boards, water containers (used to fill waterbathes), stainer containers, ice pans (all of the mentioned have been thoroughly bleached) -Tap water and DI water (all water supplies have been tested by sending a sample to micro...there was no bacteria or yeast detected) -Stay-on used as a slide adhesive (smeared some on a slide and stained it) -Bad batch of slides (we have had contamination on two different slide sources (vendors)) Does anybody have any ideas? What am I missing? Any suggestions would be much appreciated. Melissa Jans University of Iowa Hospitals and Clinics __________________________________________________ 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