From histology.bc <@t> shaw.ca Mon Aug 1 00:31:43 2005 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Excessive background staining with Millers elastic stain References: <000001c5962c$6b43cca0$e46b140a@ITP36161> Message-ID: <42EDB3BF.3030304@shaw.ca> Hi Lachlan, I believe the problem you are having is due to the presence of very similar reactive groups in both cartilage and elastic fibres, Elastin (the protein ) is heavily sulphated with an abundance of disulphide groups (hence the yellowish appearance of elastic fibres when viewed unstained or even macroscopically). When you oxidize the sections in potassium permanganate, you are converting the disulphides to sulphates, the sulphates react with Miller's elastin stain. Miller's stain is essentially as modification of aldeyde fuchsin which iis a well known method for demonstrating sulphated groups in mucopolysaccharides, etc. The extracellular matrix of cartilage consists of chondroitin sulphate (plus a few other assorted bits of connective tissue). These groups will react quite strongly with Miller's stain solution. So unfortunately, I think you may well be out of luck as far as improving the specificity of the staining reactions. The same low specificity reactions occur with most other elastin staining methods when they are used on cartilage, the reactive groups present in both locations stain with the dye in use. I cannot think of a method that stains only elastin and not cartilage matrix. Thinner sections may give you a better chance of distinguishing one from the other. Try sections at several thinner settings and see what the outcome is. Maybe there is a thickness at which you can see sufficient enough detail in the elastic fibres without the cartilage obscuring it. Paul Bradbury, Kamloops, BC Canada Lachlan Smith wrote: >Dear Subscribers > >I am staining cartilage using Millers elastic fibre stain, and while elastic >fibres are staining well, there is a lot of blue background staining which >makes thresholding for histoquantitation almost impossible. My sections are >30 >micron paraffin embedded. I am also oxidising with 0.5% aqueous potassium >permanaganate and bleaching with 2% oxalic acid. > >Any suggestings for reducing this background staining would be greatly >appreciated. > >Kind regards, > >Lachlan Smith >Institute of Medical and Veterinary Science >Adelaide, Australia > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From Kemlo.Rogerson <@t> elht.nhs.uk Mon Aug 1 05:06:44 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] New Lab Design Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4C0@elht-exch1.xelht.nhs.uk> AFOS (or something similar), formalin extraction for grossing. Includes a downdraught table and formalin making up facilities. AFOS formalin extraction cabinets for stored samples in formalin. Be careful over height of benches; some are for sitting down carrying out work, others for cutting sections using a microtome. Nothing worse than a microtome that's too high or too low. You need solvent extraction hoods for your processing machines, a suitable flammable store with antiflash lighting etc., The best lighting to get it that stuff they sell that goes in Offices; it's expensive but gives you a good colour temperature with few shadows. If you can see, stop people asphyxiating on formalin and alcohol, stop the place blowing up and have benches the right height that people can work, nowt else much to do. Oh yes, coffee machine, clean area and a Mars Bar vending machine. Ergo, all done! -----Original Message----- From: Breeden, Sara [mailto:sbreeden@nmda.nmsu.edu] Sent: 29 July 2005 17:27 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New Lab Design A new histology lab (in a new building) is in the works (yipee!!). I have some definite ideas about what I'll need in my new digs, but I need more opinions! Within the next couple weeks, I will be asked by the architects for my input. This is a State veterinary lab where I do "surgicals", necropsies, standard special stains, with a rapidly expanding IHC workload. My total workload was roughly 10,000 blocks last year and I expect it to grow by 5% each year. I expect to have roughly 500 sq ft to work with in this new space. I work alone for three DVM pathologists. What would you consider absolutely essential in a new histology lab? What would you do differently if you'd had a chance? What kind of: bench space and type of surface, ventilation, lighting, hoods, haz mat storage, supply storage, windows? I only get once chance at this and I want to make it count...I have 8 more years until retirement and I'd rather not kick myself for missing something obvious. Please reply directly to me. Thank you in advance!! I rely on 'netters for great information. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ROrr <@t> enh.org Mon Aug 1 07:26:53 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Biotinylating Antibodies Message-ID: Vernon, I have been successful with a biotinylation kit from Biocare Medical. It uses a different chemistry than the Vector kits. I think the mopping reagent may be what makes the difference. Their number is 800 799 9499 ask for Hari, he's the technical sales manager. Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 From DDittus787 <@t> aol.com Mon Aug 1 07:46:13 2005 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] EGFR Message-ID: <9b.648c607f.301f7395@aol.com> Josie: Are you aware of the clone 31g7 and Allen Gowns ASCO 2005 poster. It showed that 31g7 from Zymed was best in class and selected 10-15% more patients. dana From pruegg <@t> ihctech.net Mon Aug 1 11:21:23 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] (no subject) Message-ID: <200508011621.j71GLFpf017202@chip.viawest.net> Does anyone know where I could get a service manual for an old IEC cryostat? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From pruegg <@t> ihctech.net Mon Aug 1 11:23:37 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] (no subject) In-Reply-To: <1122805089.42eca561c7451@mail.imvs.org> Message-ID: <200508011623.j71GNTpf017949@chip.viawest.net> We used picro sirrus stain for collagen fibers and viewed the collagen bundles using polarized light filter for histomorphometry. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of lachlan.smith@imvs.sa.gov.au Sent: Sunday, July 31, 2005 3:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Dear Subscribers I am staining cartilage using Millers elastic fibre stain, and while elastic fibres are staining well, there is a lot of blue background staining which makes thresholding for histoquantitation almost impossible. My sections are 30 micron paraffin embedded. I am also oxidising with 0.5% aqueous potassium permanaganate and bleaching with 2% oxalic acid. Any suggestings for reducing this background staining would be greatly appreciated. Kind regards, Lachlan Smith Institute of Medical and Veterinary Science Adelaide, Australia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Aug 1 13:06:41 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Problems of processing brain for paraffin embedding In-Reply-To: <20050730083029.26833.qmail@web52102.mail.yahoo.com> References: <20050730083029.26833.qmail@web52102.mail.yahoo.com> Message-ID: <6.0.0.22.1.20050801120504.01b73828@gemini.msu.montana.edu> Are you processing whole brain, or coronal slices? Once you indicate, I will be happy to send our protocols via private email attachement. At 02:30 AM 7/30/2005, you wrote: >Dear Members, >I have problems with paraffin processing of mouse and hamster brains and >spinal cords. I have perfused the animals with PFA 4% and I postfixed them >in the same buffer for 7-8 days and then stored in Alc 70% (up to now). >After sectioning, tissues wrinkle in a way that it is torn after >flattenning (on a slide warmer), therefore the quality is not good at all. > >Can anyone give me hints to overcome the problem. A detailted protocol of >paraffin processing would be great. > >Regard >A.P. Tafreshi 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 sbreeden <@t> nmda.nmsu.edu Mon Aug 1 13:33:25 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] New Lab Design Message-ID: Just a quick note to thank all of you that sent me your ideas/comments/input on what to put into a new histo lab. These ideas have been duly noted and will be part of My Grand Plan. There were some meaty ideas and some real sparklers! Your help is invaluable -- and if you've thought of anything else since last week, let me know. Thanks again! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Monday, August 01, 2005 11:02 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 21, Issue 1 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Excessive background staining with Millers elastic stain (Lachlan Smith) 2. staining for iron in microglia (Sharon Cooperman) 3. Re: Excessive background staining with Millers elastic stain (Paul Bradbury) 4. RE: New Lab Design (Rogerson Kemlo (ELHT) Pathology) 5. Biotinylating Antibodies (Orr, Rebecca) 6. Re: EGFR (DDittus787@aol.com) 7. (no subject) (Patsy Ruegg) 8. RE: (no subject) (Patsy Ruegg) ---------------------------------------------------------------------- Message: 1 Date: Mon, 1 Aug 2005 09:33:17 +0930 From: "Lachlan Smith" Subject: [Histonet] Excessive background staining with Millers elastic stain To: Message-ID: <000001c5962c$6b43cca0$e46b140a@ITP36161> Content-Type: text/plain; charset="iso-8859-1" Dear Subscribers I am staining cartilage using Millers elastic fibre stain, and while elastic fibres are staining well, there is a lot of blue background staining which makes thresholding for histoquantitation almost impossible. My sections are 30 micron paraffin embedded. I am also oxidising with 0.5% aqueous potassium permanaganate and bleaching with 2% oxalic acid. Any suggestings for reducing this background staining would be greatly appreciated. Kind regards, Lachlan Smith Institute of Medical and Veterinary Science Adelaide, Australia ------------------------------ Message: 2 Date: Sun, 31 Jul 2005 21:39:40 -0400 From: Sharon Cooperman Subject: [Histonet] staining for iron in microglia To: Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Dear Histonetters, I have a semi-scientific question: macrophages in almost all tissues can be stained for iron with Perl's stain to give a blue color - is this also true for microglia? If not, is there any other way to see how much iron is in microglia (DAB enhanced Perl's or some other method)? Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 ------------------------------ Message: 3 Date: Sun, 31 Jul 2005 22:31:43 -0700 From: Paul Bradbury Subject: Re: [Histonet] Excessive background staining with Millers elastic stain To: Lachlan Smith , HistoNet Server Message-ID: <42EDB3BF.3030304@shaw.ca> Content-Type: text/plain; format=flowed; charset=us-ascii Hi Lachlan, I believe the problem you are having is due to the presence of very similar reactive groups in both cartilage and elastic fibres, Elastin (the protein ) is heavily sulphated with an abundance of disulphide groups (hence the yellowish appearance of elastic fibres when viewed unstained or even macroscopically). When you oxidize the sections in potassium permanganate, you are converting the disulphides to sulphates, the sulphates react with Miller's elastin stain. Miller's stain is essentially as modification of aldeyde fuchsin which iis a well known method for demonstrating sulphated groups in mucopolysaccharides, etc. The extracellular matrix of cartilage consists of chondroitin sulphate (plus a few other assorted bits of connective tissue). These groups will react quite strongly with Miller's stain solution. So unfortunately, I think you may well be out of luck as far as improving the specificity of the staining reactions. The same low specificity reactions occur with most other elastin staining methods when they are used on cartilage, the reactive groups present in both locations stain with the dye in use. I cannot think of a method that stains only elastin and not cartilage matrix. Thinner sections may give you a better chance of distinguishing one from the other. Try sections at several thinner settings and see what the outcome is. Maybe there is a thickness at which you can see sufficient enough detail in the elastic fibres without the cartilage obscuring it. Paul Bradbury, Kamloops, BC Canada Lachlan Smith wrote: >Dear Subscribers > >I am staining cartilage using Millers elastic fibre stain, and while >elastic fibres are staining well, there is a lot of blue background >staining which makes thresholding for histoquantitation almost >impossible. My sections are 30 micron paraffin embedded. I am also >oxidising with 0.5% aqueous potassium permanaganate and bleaching with >2% oxalic acid. > >Any suggestings for reducing this background staining would be greatly >appreciated. > >Kind regards, > >Lachlan Smith >Institute of Medical and Veterinary Science Adelaide, Australia > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ Message: 4 Date: Mon, 1 Aug 2005 11:06:44 +0100 From: "Rogerson Kemlo (ELHT) Pathology" Subject: RE: [Histonet] New Lab Design To: "Breeden, Sara" , Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4C0@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" AFOS (or something similar), formalin extraction for grossing. Includes a downdraught table and formalin making up facilities. AFOS formalin extraction cabinets for stored samples in formalin. Be careful over height of benches; some are for sitting down carrying out work, others for cutting sections using a microtome. Nothing worse than a microtome that's too high or too low. You need solvent extraction hoods for your processing machines, a suitable flammable store with antiflash lighting etc., The best lighting to get it that stuff they sell that goes in Offices; it's expensive but gives you a good colour temperature with few shadows. If you can see, stop people asphyxiating on formalin and alcohol, stop the place blowing up and have benches the right height that people can work, nowt else much to do. Oh yes, coffee machine, clean area and a Mars Bar vending machine. Ergo, all done! -----Original Message----- From: Breeden, Sara [mailto:sbreeden@nmda.nmsu.edu] Sent: 29 July 2005 17:27 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New Lab Design A new histology lab (in a new building) is in the works (yipee!!). I have some definite ideas about what I'll need in my new digs, but I need more opinions! Within the next couple weeks, I will be asked by the architects for my input. This is a State veterinary lab where I do "surgicals", necropsies, standard special stains, with a rapidly expanding IHC workload. My total workload was roughly 10,000 blocks last year and I expect it to grow by 5% each year. I expect to have roughly 500 sq ft to work with in this new space. I work alone for three DVM pathologists. What would you consider absolutely essential in a new histology lab? What would you do differently if you'd had a chance? What kind of: bench space and type of surface, ventilation, lighting, hoods, haz mat storage, supply storage, windows? I only get once chance at this and I want to make it count...I have 8 more years until retirement and I'd rather not kick myself for missing something obvious. Please reply directly to me. Thank you in advance!! I rely on 'netters for great information. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 1 Aug 2005 07:26:53 -0500 From: "Orr, Rebecca" Subject: [Histonet] Biotinylating Antibodies To: Message-ID: Content-Type: text/plain; charset="us-ascii" Vernon, I have been successful with a biotinylation kit from Biocare Medical. It uses a different chemistry than the Vector kits. I think the mopping reagent may be what makes the difference. Their number is 800 799 9499 ask for Hari, he's the technical sales manager. Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 ------------------------------ Message: 6 Date: Mon, 1 Aug 2005 08:46:13 EDT From: DDittus787@aol.com Subject: Re: [Histonet] EGFR To: JosefaNava@texashealth.org, histonet@pathology.swmed.edu Message-ID: <9b.648c607f.301f7395@aol.com> Content-Type: text/plain; charset="US-ASCII" Josie: Are you aware of the clone 31g7 and Allen Gowns ASCO 2005 poster. It showed that 31g7 from Zymed was best in class and selected 10-15% more patients. dana ------------------------------ Message: 7 Date: Mon, 1 Aug 2005 10:21:23 -0600 From: "Patsy Ruegg" Subject: [Histonet] (no subject) To: "'Histonet'" Message-ID: <200508011621.j71GLFpf017202@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Does anyone know where I could get a service manual for an old IEC cryostat? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. ------------------------------ Message: 8 Date: Mon, 1 Aug 2005 10:23:37 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] (no subject) To: , Message-ID: <200508011623.j71GNTpf017949@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" We used picro sirrus stain for collagen fibers and viewed the collagen bundles using polarized light filter for histomorphometry. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of lachlan.smith@imvs.sa.gov.au Sent: Sunday, July 31, 2005 3:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Dear Subscribers I am staining cartilage using Millers elastic fibre stain, and while elastic fibres are staining well, there is a lot of blue background staining which makes thresholding for histoquantitation almost impossible. My sections are 30 micron paraffin embedded. I am also oxidising with 0.5% aqueous potassium permanaganate and bleaching with 2% oxalic acid. Any suggestings for reducing this background staining would be greatly appreciated. Kind regards, Lachlan Smith Institute of Medical and Veterinary Science Adelaide, Australia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 21, Issue 1 *************************************** From sbreeden <@t> nmda.nmsu.edu Mon Aug 1 14:17:21 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Results of "New Lab Ideas" Poll Message-ID: And with many thanks to all of you that had input, herewithin is a list of the thoughts. And, might I add, this is just one reason I think this medium (Histonet) is absolutely necessary!! Lookie what I found out!!... Large SINKS - and more than one; sink in HOOD for specials, IHC, etc.; ergonomically correct SEATING and COUNTER HEIGHT for task; proper VENTILATION and AIR DIRECTION (as in no airflow above microtome/water bath, but proper ventilation for fume removal and room temperature control); BRIGHT WHITE LIGHTING in task areas; VENT (ambient air pressure?) or similar over embedding area/processor (for heat removal); DEDICATED CIRCUITS for essential equipment and BACKUP POWER UNITS for same; D.I. WATER; separate HAZMAT STORAGE and STORAGE for acids and bases (separately); STORAGE for supplies/inventory (larger than you think you'll ever need); EYEWASH/SHOWER; TEXTURED FLOOR; 30" deep COUNTERS; rounded COUNTERTOP edges; COUNTERTOPS that don't stain or corrode (stainless steel not recommended); no GLASS-INSERT DOORS (visible clutter); overhead VENTS w/snorkels; and appropriate WORKFLOW planning. Of course, this prompted a few "needs" for my particular space, which I will list just for informational purposes. But it may jog a thought for someone in a similar position: HAZMAT storage out of work area or in an area that's near the need (i.e., processor); trash cans out of walkway (!); area for slide return (post-pathologist); locking cabinets and drawers (for blades, knives, Secret Histology Stuff, etc.); and under-cabinet lighting. I'm sure the list will grow. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From Jane_Casey <@t> brown.edu Mon Aug 1 14:22:58 2005 From: Jane_Casey <@t> brown.edu (Casey, Jane) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Embedding human cartilage/joint tissue in plastic Message-ID: <471F15E271937A4DA89AC12211DDAD980266E9E3@MAIL2.AD.Brown.Edu> We are trying to use histology to validate our method of measuring cartilage thickness on wrist bones using ?CT scans. Initially we tried fixing a trapezoid fragment in formalin, de-mineralizing it, and embedding it in paraffin before slicing it into 5 ?m sections. This method led to non-uniform shrinkage of the tissue and clearly visible artifact. We would like to try a method, perhaps embedment in plastic, which will yield a more accurate measure of cartilage thickness. Does anyone have any suggestions? Thank you for your suggestions. Jane Casey Department of Orthopaedics Brown Medical School / Rhode Island Hospital Providence, RI 02912 Jane_Casey@Brown.edu From eca9 <@t> georgetown.edu Mon Aug 1 15:25:34 2005 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] mouse anti-Her2 Message-ID: <42EE853E.2010103@georgetown.edu> Hello Everyone, I just wanted to ask about a mouse anti-Her2 clone CB11 antibody that I have bought from Zymed. It says the optimal dilutions are to be determined by each lab but I just want to know if someone could tell me which dilution they use. Also do you use an antigen retrieval method and which detection method do you use? Thank you Eva Georgetown University From gcallis <@t> montana.edu Mon Aug 1 15:28:34 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:24 2005 Subject: Avoiding shrinkage Re: [Histonet] Embedding human cartilage/joint tissue in plastic In-Reply-To: <471F15E271937A4DA89AC12211DDAD980266E9E3@MAIL2.AD.Brown.Ed u> References: <471F15E271937A4DA89AC12211DDAD980266E9E3@MAIL2.AD.Brown.Edu> Message-ID: <6.0.0.22.1.20050801141209.01b3ac10@gemini.msu.montana.edu> A clever, simple use of low voltage xray machines i.e FAXITRON can help you out. We did specimen radiographs with a FAXITRON, and underexposed the sample to enhance soft tissues i.e tumors and cartilage. These will be like "contact" photographs, showing no shrinkage of the soft tissue components. After fixation DO not decalcify, radiograph the sample then decalcify (by the way, xray is the most sensitive decalcification endpoint determination) and do another radiograph at the same initial exposure. You can do thicker slabs even 5 mm thick but make sure you are consistent with thickness of slab, a good bone saw will do this - Buehler Isomet or the MarMed saw, a lovely device. The bones can be NBF fixed but you avoid all dehydration/clearing and heat of paraffin which will cause shrinkage. If you have to know exact edge of cartilage, a radioopaque substance can be painted on surface of cartilage to define that sharp edge. There is a publication where a group tested the amount of shrinkage you get with both paraffin and plastic processing, results were interesting in that BOTH methods caused as much as 20% shrinkage, possibly more of the hard tissue, so you will not avoid shrinkage with plastic embedding methods involving alcohols or any other solvent to remove water. Since cartilage has a high water content and by doing specimen radiography, you can avoid alcohols, clearing agents and heat of paraffin that cause shrinkage by water removal. At 01:22 PM 8/1/2005, you wrote: > > >We are trying to use histology to validate our method of measuring >cartilage thickness on wrist bones using ?CT scans. Initially we tried >fixing a trapezoid fragment in formalin, de-mineralizing it, and embedding >it in paraffin before slicing it into 5 ?m sections. This method led to >non-uniform shrinkage of the tissue and clearly visible artifact. We would >like to try a method, perhaps embedment in plastic, which will yield a >more accurate measure of cartilage thickness. Does anyone have any >suggestions? > >Thank you for your suggestions. > > >Jane Casey >Department of Orthopaedics >Brown Medical School / Rhode Island Hospital >Providence, RI 02912 >Jane_Casey@Brown.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 RCazares <@t> schosp.org Mon Aug 1 16:09:48 2005 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Histo tech position available Message-ID: <913FAC2B773C19488E26AE6572180FA5029A992C@exch01.schosp.org> Hi all, We have a position open for a histo tech at Swedish Covenant Hospital in Chicago. The position is alternate Saturdays (8 hour day) and fill in for vacations if possible. There is also opportunity to pick up extra hours during the week on evenings but this is optional. There will also be a full time position open in December when one of my FT techs retires. Interested techs (certified) can call or fax their resume; Ruth Cazares Department of Pathology Phone: 773 878-8200 ext-5190 Fax: 773 271-1501 (to my attention) *** 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 jkiernan <@t> uwo.ca Mon Aug 1 23:16:09 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] staining for iron in microglia References: Message-ID: <42EEF389.A65BF4AC@uwo.ca> Sharon, Your question is 100% scientific, not semi-scientific! You concisely provide an introduction, expand it into a testable hypothesis, and ask if the test has been done. The answer is "Yes, but ..." Iron is seen in phagocytic cells in various pathological conditions of the brain. Its presence in the frontal cortex (seen with the regular Perls method) is a classical feature of advanced syphilis (general paralysis of the insane). There are some cells in the normal rat's hypothalamus that have a variety of histochemical peculiarities including being Perls-positive, but these cells are astrocytes (Young & McKenzie, 2004). The DAB-enhanced Perls method shows non-haem iron in microglia, oligodendroglia and neurons. Research in this field, from several labs, was published in the late 1980s and early 1990s, about 10 years after the publication of the DAB amplification procedure by Nguyen-Legros et al in 1980. A few references are given at the end of this message. They are ones that I've seen and made notes on; there will be many more, findable by way of PubMed, Web of Science or Scopus. At NIH all those resources and more will be available to you. Another iron-related approach to microglia has been immunostaining for ferritin (Kaneko et al 1989). There are some lectins with affinity for microglia that can provide impressive pictures when correctly applied. For the old and bold there are the original silver stains, developed by Del Rio Hortega and perfected by Penfield and Cone in the 1920s. Summary. Sensitive methods for iron do not provide specific staining of resident, activated or immigrant microglial cells. The classical Perls method can detect microglia that have collected iron in sites of injury or disease. Iron is also present in macroglia and neurons, and is stainable in these cells if the sensitivity of the Perls prussian blue method is enhanced. References. Gerber MR, Connor JR (1989) Do oligodendrocytes mediate iron regulation in the human brain? Ann. Neurol. 26: 95-98. Kaneko Y, Kitamoto T, Tateishi J, Yamaguchi K (1989) Ferritin immunohistochemistry as a marker for microglia. Acta Neuropathol. (Berl.) 79: 129-136. Morris CM, Candy JM, Oakley AE, Bloxham CA, Edwardson JA (1992) Histochemical distribution of non-haem iron in the human brain. Acta Anat. 144: 235-257. Connor JR, Pavlick G, Karli D, Menzies SL, Palmer C (1995) A histochemical study of iron-positive cells in the developing rat brain. J. Comp. Neurol. 355: 111-123. Palmer C, Menzies SL, Roberts RL, Pavlick G, Connor JR (1999) Changes in iron histochemistry after hypoxic-ischemic brain injury in the neonatal rat. J. Neurosci. Res. 56: 60-71. Young JK, McKenzie JC (2004) GLUT2 immunoreactivity in Gomori-positive astrocytes of the hypothalamus. J. Histochem. Cytochem. 52: 1519-1524. John Kiernan Anatomy, UWO London, Canada --------------------------------------- Sharon Cooperman wrote: > > Dear Histonetters, > > I have a semi-scientific question: macrophages in almost all tissues > can be stained for iron with Perl's stain to give a blue color - is > this also true for microglia? If not, is there any other way to see > how much iron is in microglia (DAB enhanced Perl's or some other > method)? > > Thanks, > Sharon > -- > Sharon Cooperman > NIH, NICHD, CBMB 301.435-8417 > Building 18T, room 101 301.402-0078 fax > Bethesda, MD 20892 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Tue Aug 2 02:41:07 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] RE: New lab design Message-ID: One thing you need to over-estimate on is block and slide storage, and if you are not on a ground floor, they weigh a ton so you'll need some additional support system. Cheers Jacqui Malam DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From peter_bannister <@t> hotmail.co.uk Tue Aug 2 06:01:29 2005 From: peter_bannister <@t> hotmail.co.uk (Peter Bannister) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Does anyone have a spare tissue processor that is getting in the way? Message-ID: Hello again Histonetters, I realise that this is a slightly cheeky message, but I need to ask if any of you (particularly those based in the UK) have an old carousel style tissue processor taking up valuable storage space in your lab? We are currently setting up a new research lab here in london that will have a relatively low specimen throughput and a rather limited equipment budget. We would therefore be eternally grateful if you would allow us to help you dispose of your old kit! We will of course collect and I will personally chip in towards your christmas party. Yes it's cheeky - but 'if you don't ask, you don't get!' If you can help, please contact me directly. Many thanks for your time. Peter Bannister _________________________________________________________________ It's fast, it's easy and it's free. Get MSN Messenger 7.0 today! http://messenger.msn.co.uk From jluis.palazon <@t> icman.csic.es Tue Aug 2 06:36:04 2005 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] ANTI-MYOSIN AND ANTIRENIN Message-ID: <20050802113604.74D8A89C1F5@perceval.uca.es> Dear List-members I am finishing my doctoral thesis and would like to demonstrate renin in an aglomerular teleost kidney, smooth muscle in the iris, and muscle in the bulbus arteriosus. I know that this can be done inmunohistologically but my lab does not work in inmunohistochemistry so I do not have the antibodies. I would like to know if there is someone of our list here in Spain (hospital, University) that can do this kind of inmunos and the posibility of doing my samples with them or maybe sending the samples and paying for the inmunos. thanks in advance Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From jluis.palazon <@t> icman.csic.es Tue Aug 2 06:37:43 2005 From: jluis.palazon <@t> icman.csic.es (Jose Luis Palazon Fernandez) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] retina neurons Message-ID: <20050802113743.3F3E989C5A1@perceval.uca.es> Dear list fellows is there any simple stain that I can use to differentiate between amacrine, horizontal and bipolar cells in the retina. I have tried using H&E but as I have not experience in this kind of tissue I dont know how to differentiate between them, mainly between amacrine and bipolar cells. thanks in advance jose luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es From cdemarinis <@t> SARATOGACARE.ORG Tue Aug 2 06:43:35 2005 From: cdemarinis <@t> SARATOGACARE.ORG (Demarinis, Carolyn) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] cassette labeller Message-ID: We are interested in obtaining a cassette labeller. Does anyone have any recommendations? We are a Meditech hospital. *************************************************************************************** CONFIDENTIALITY NOTICE: This e-mail communication and any attachmentsmay 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. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. From emerald_lake77 <@t> yahoo.com Tue Aug 2 08:05:22 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Elastic Trichrome - any good protocol out there not involving microwave??? Message-ID: <20050802130522.4981.qmail@web31710.mail.mud.yahoo.com> Hello all, I am in need of a good protocol for a combination elastic and trichrome stain that doesn't involve microwave. Thanks. Gustave Hebert Scientist II Wyeth Research Cambridge MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From PMonfils <@t> Lifespan.org Tue Aug 2 08:52:22 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Elastic Trichrome - any good protocol out there no t involving microwave??? Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171759D@lsexch.lsmaster.lifespan.org> I often combine the Verhoeff Elastin Stain with the Gomori Trichrome. I do the Bouin's treatment required for the trichrome procedure first. Then I do the complete Verhoeff stain, eliminatiing the final Van Gieson counterstain. Then I apply the one-step Gomori Trichrome. It is quite simple and gives excellent results. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of GT > Hebert > Sent: Tuesday, August 2, 2005 6:05 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Elastic Trichrome - any good protocol out there > not involving microwave??? > > Hello all, > > I am in need of a good protocol for a combination elastic and trichrome > stain that doesn't involve microwave. Thanks. > > Gustave Hebert > Scientist II > Wyeth Research > Cambridge MA > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From bhewlett <@t> cogeco.ca Tue Aug 2 09:20:58 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Elastic Trichrome - any good protocol out there not involving microwave??? References: <09C945920A6B654199F7A58A1D7D1FDE0171759D@lsexch.lsmaster.lifespan.org> Message-ID: <01a301c5976d$67bdd250$6400a8c0@mainbox> Paul, The Van Gieson counterstain IS a trichrome! It also has the advantage of not requiring the Bouin pretreatment. An excellent alternative to Verhoff, is the Miller elastic stain which also incorporates the Van Gieson. Bryan ----- Original Message ----- From: "Monfils, Paul" To: Sent: Tuesday, August 02, 2005 9:52 AM Subject: RE: [Histonet] Elastic Trichrome - any good protocol out there not involving microwave??? >I often combine the Verhoeff Elastin Stain with the Gomori Trichrome. I do > the Bouin's treatment required for the trichrome procedure first. Then I > do > the complete Verhoeff stain, eliminatiing the final Van Gieson > counterstain. > Then I apply the one-step Gomori Trichrome. It is quite simple and gives > excellent results. > > Paul M. > >> ---------- >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of GT >> Hebert >> Sent: Tuesday, August 2, 2005 6:05 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Elastic Trichrome - any good protocol out there >> not involving microwave??? >> >> Hello all, >> >> I am in need of a good protocol for a combination elastic and trichrome >> stain that doesn't involve microwave. Thanks. >> >> Gustave Hebert >> Scientist II >> Wyeth Research >> Cambridge MA >> >> __________________________________________________ >> Do You Yahoo!? >> Tired of spam? Yahoo! Mail has the best spam protection around >> http://mail.yahoo.com >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Tue Aug 2 09:31:15 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Elastic Trichrome - any good protocol out there no t involving microwave??? Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB434C@fh2k093.fhmis.net> Generally, we find that the steps are so short anyway that we don't use the microwave on the masson's trichrome-except for the Bouin's Solution and even then we try not to because the solution becomes compromised. ---Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Monfils, Paul; histonet@lists.utsouthwestern.edu Sent: 8/2/2005 10:20 AM Subject: Re: [Histonet] Elastic Trichrome - any good protocol out there not involving microwave??? Paul, The Van Gieson counterstain IS a trichrome! It also has the advantage of not requiring the Bouin pretreatment. An excellent alternative to Verhoff, is the Miller elastic stain which also incorporates the Van Gieson. Bryan ----- Original Message ----- From: "Monfils, Paul" To: Sent: Tuesday, August 02, 2005 9:52 AM Subject: RE: [Histonet] Elastic Trichrome - any good protocol out there not involving microwave??? >I often combine the Verhoeff Elastin Stain with the Gomori Trichrome. I do > the Bouin's treatment required for the trichrome procedure first. Then I > do > the complete Verhoeff stain, eliminatiing the final Van Gieson > counterstain. > Then I apply the one-step Gomori Trichrome. It is quite simple and gives > excellent results. > > Paul M. > >> ---------- >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of GT >> Hebert >> Sent: Tuesday, August 2, 2005 6:05 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Elastic Trichrome - any good protocol out there >> not involving microwave??? >> >> Hello all, >> >> I am in need of a good protocol for a combination elastic and trichrome >> stain that doesn't involve microwave. Thanks. >> >> Gustave Hebert >> Scientist II >> Wyeth Research >> Cambridge MA >> >> __________________________________________________ >> Do You Yahoo!? >> Tired of spam? Yahoo! Mail has the best spam protection around >> http://mail.yahoo.com >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bjrenquist <@t> ucdavis.edu Tue Aug 2 11:03:42 2005 From: bjrenquist <@t> ucdavis.edu (Ben) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Whole Slide Imaging Message-ID: <200508021603.j72G3gUj002726@pop19.ucdavis.edu> Hello All, I'm interested in whole slide imaging for some research work. I was hoping that anyone who has worked with it could give me their opinion on the equipment they use, the important features (by the way I'm looking to count nuclei), or suggest a good place to get this done for a decent price. If this message is against the rules of histonet, I apologize, but if not I appreciate your help. Ben Renquist UC Davis From lrichey <@t> u.washington.edu Tue Aug 2 12:50:21 2005 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Elastic Trichrome - any good protocol out there not involving microwave??? In-Reply-To: <20050802130522.4981.qmail@web31710.mail.mud.yahoo.com> References: <20050802130522.4981.qmail@web31710.mail.mud.yahoo.com> Message-ID: <42EFB25D.6020705@u.washington.edu> Movats might be a good one. It takes about 3 hours and does not need a microwave. GT Hebert wrote: >Hello all, > >I am in need of a good protocol for a combination elastic and trichrome stain that doesn't involve microwave. Thanks. > >Gustave Hebert >Scientist II >Wyeth Research >Cambridge MA > >__________________________________________________ >Do You Yahoo!? >Tired of spam? Yahoo! Mail has the best spam protection around >http://mail.yahoo.com >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From emry <@t> u.washington.edu Tue Aug 2 13:23:34 2005 From: emry <@t> u.washington.edu (Trisha Emry) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] nasal septum/Prefer Message-ID: We are still having trouble keeping nasal septums on the slides doing brdu. Now we are asking if the problem could be the fixative. Is anyone doing cartilage using Prefer? Any problems or special techniques needed to make it work? Thanks, Trisha Seattle From TillRenee <@t> uams.edu Tue Aug 2 13:45:38 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] rat on rat Message-ID: Any suggestions? The antibody is a mouse anti-rat and it is to be used on rat tissue. Does that present the same problems like mouse on mouse? If so, what do you can you do about it? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 From pwg1 <@t> cdc.gov Tue Aug 2 13:50:13 2005 From: pwg1 <@t> cdc.gov (Greer, Patricia) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Job opportunity in Atlanta Message-ID: The following is an excerpt from a job announcement for Biologist and/or Medical Technologist that will be posted sometime this week for a position in the Infectious Disease Pathology Laboratory at The Centers for Disease Control and Prevention in Atlanta: The incumbent processes animal and human tissues for histologic examination utilizing routine (hematoxylin and eosin) and special staining procedures for the identification of infectious agents and tissue components; processes tissue sections for immunohistochemical (immunofluorescence and immunoenzymatic) and in situ hybridization studies. Performs specialized pathology techniques to detect the presence of infectious disease agent antigens, proteins and nucleic acids in clinical/research tissue.... KNOWLEDGE, SKILLS AND ABILITIES (KSAs): KSAs are the specific characteristics that applicants should possess in order to perform the major duties of the position. Applications should address the specific KSAs as part of your application. KSAs identified as (M) are considered critical to the position and are considered to be mandatory for qualifications. KSAs identified as (D) are considered to be desirable. FAILURE TO ADDRESS KSAs MAY RESULT IN A LOWER RATING. 1. Please explain how you meet the qualification requirements and specialized experience for this position as defined in the vacancy announcement. 2. Ability to perform routine and specialized histology procedures. (M) 3. Skill in performing specialized immunohhistochemical and molecular pathology techniques. (M) 4. Ability to process and track a large number of biologic samples according to standard protocols. (M) 5. Skill in the use of laboratory microcomputers and software. (D) 6. Knowledge of biosafety and chemical safety relevant to common pathology laboratory procedures. (D) For each of the above, give examples of how you gained the knowledge, skill, or ability and the dates of such experience and education. Click here for instruction on how to respond to KSA's. You can search for the position/positions and fill out the on-line application at: http://www2.cdc.gov/hrmo/vsearch.asp Pat Greer Infectious Disease Pathology Activity Centers for Disease Control and Prevention Mail Stop G-32 Atlanta, GA 30333 404-639-2811 From portera203 <@t> yahoo.com Tue Aug 2 14:34:58 2005 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] rat on rat In-Reply-To: Message-ID: <20050802193458.70000.qmail@web34113.mail.mud.yahoo.com> Renee' - the host of the primary is a mouse if I am reading your email right...you should be able to use a secondary against mouse that is rat absorbed. We do this quite often and have minimal problems. "Till, Renee" wrote:Any suggestions? The antibody is a mouse anti-rat and it is to be used on rat tissue. Does that present the same problems like mouse on mouse? If so, what do you can you do about it? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Amy S.Porter, HT(ASCP), QIHC Laboratory Supervisor Michigan State University - Department of Physiology Division of Human Pathology Investigative Histopathology Laboratory 2201 Biomedical Physical Sciences Room 2133 East Lansing, MI 48824-3320 Ph: 517-355-6475 ext 1480/1481 Fax: 517-432-1368 portera@msu.edu / www.humanpathology.msu.edu __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From TillRenee <@t> uams.edu Tue Aug 2 14:49:10 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] ooops, rat on rat Message-ID: Okay, I knew I'd say that wrong. It is a rat anti-mouse antibody to be used on rat tissues. How would you handle that? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 From bwhitaker <@t> brownpathology.com Tue Aug 2 15:03:21 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] slide disposal Message-ID: <000301c5979d$3b7721d0$3601a8c0@brownpathology.net> Hi All, Can some of you give me ideas of how you handle old slide disposal? Particularly those of you in smaller, private labs. I have about 20 years worth of slides that I need to dispose of. If I have to send them with my biohazardous waste, I can, but it seems like an unnecessary expense. Currently that is what the procedure says that we do, but is it really necessary? Maybe someone out there would like to build a glass house : ) Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 From gcallis <@t> montana.edu Tue Aug 2 15:13:10 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Re: rat on rat In-Reply-To: References: Message-ID: <6.0.0.22.1.20050802140415.01b3b210@gemini.msu.montana.edu> No problems - but you would have rat on rat problems if you were using a rat host primary i.e. Rat antiRat primary. You can use a donkey or goat antiMouse secondary adsorbed to rat tissues, preferably an F(ab')2 frag of IgG - Jackson has excellent secondaries for this. We used a goat antiMouse isotype i.e IgG1, 2, etc -biotin conjugate from Southern Biotechnology. The latter was very clean and I strongly recommend these. Just do normal serum blocking carefully - you could even add 1% rat serum to the normal serum block with either goat or donkey serum but I don't think that will be needed. At 12:45 PM 8/2/2005, you wrote: >Any suggestions? The antibody is a mouse anti-rat and it is to be used >on rat tissue. Does that present the same problems like mouse on mouse? >If so, what do you can you do about it? > > > > > >Renee' Till, HT > > > >Arkansas Childrens Nutrition Center > >1120 Marshall > >Little Rock, AR > >72202 > > > >(501) 364-2774 > > > >_______________________________________________ >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 PMonfils <@t> Lifespan.org Tue Aug 2 15:15:53 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Whole Slide Imaging Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171759E@lsexch.lsmaster.lifespan.org> Hi Ben, For imaging of whole sections up to about 1 cm in length we use a Nikon microscope with a 1x objective. For whole imaging of larger sections we use a Polaroid Sprint Scan slide scanner, which does a great job. It is actually designed for scanning photographic slides, but has an adaptor which allows direct scanning of microscope slides at resolutions up to 2500 DPI. Both instruments capture images directly to the computer. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ben > Sent: Tuesday, August 2, 2005 9:03 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Whole Slide Imaging > > Hello All, > > > > I'm interested in whole slide imaging for some research work. I was > hoping > that anyone who has worked with it could give me their opinion on the > equipment they use, the important features (by the way I'm looking to > count > nuclei), or suggest a good place to get this done for a decent price. If > this message is against the rules of histonet, I apologize, but if not I > appreciate your help. > > > > Ben Renquist > > UC Davis > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gcallis <@t> montana.edu Tue Aug 2 15:21:17 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Re: ooops, rat on rat In-Reply-To: References: Message-ID: <6.0.0.22.1.20050802141439.01b3b210@gemini.msu.montana.edu> You could use a biotinylated primary, as in the DAKO ARK methodology, eliminate the secondary to avoid having some "host" antiRat detect not only the rat primary, but also bind to rat tissue, then come back with a Strepavidin-HRP or AP. Sometimes you have to dilute out the primary more - I am not sure if anyone has a rat on rat kit, try Scytek out of Logan UT, you might get lucky on a kit. At 01:49 PM 8/2/2005, you wrote: >Okay, I knew I'd say that wrong. It is a rat anti-mouse antibody to be >used on rat tissues. How would you handle that? > > > >Renee' Till, HT > > > >Arkansas Childrens Nutrition Center > >1120 Marshall > >Little Rock, AR > >72202 > > > >(501) 364-2774 > > > >_______________________________________________ >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 sthevar <@t> bu.edu Tue Aug 2 17:21:20 2005 From: sthevar <@t> bu.edu (Sandy Thevarkunnel) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Kainate Receptor radioligand Message-ID: <000001c597b0$82a11440$6b83299b@Rita> Hi, I've been searching all over to find [3H] Kainate (which isn't offered by New England Nuclear anymore) or another type of ligand that is commercially available that binds to kainate receptors in human/primate cerebellum. Thanks in advance, Sandy Graduate Student From clarissabush <@t> sbcglobal.net Tue Aug 2 18:19:19 2005 From: clarissabush <@t> sbcglobal.net (CM Bush) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Shrinkage of frozen fixed brain tissue mounted on slides Message-ID: <20050802231919.58798.qmail@web80305.mail.yahoo.com> Dear Histonet, I have been cutting frozen fixed human brain tissue, using a freezing stage on a rotary microtome. The tissue has been very well fixed in 10%NBF, and cryoprotected with sucrose. I cut the sections at 50 um, floated the sections in PBS, mounted them on slides, and allowed them to dry for 48hrs and then Nissl stained. Everything looks nice except that the sections have shrunk down to 20um as measured using a stereology set up. This is very sad. I guess we will just have to cut thicker sections... My question is, does this sound like the amount of shrinkage to expect with frozen fixed brain tissue mounted onto slides? It makes sense that there would be some change due to expansion of freezing water, and then drying on the slide but greater than 50% change? Thank you in advance, for your help. CM Bush From Eric <@t> ategra.com Tue Aug 2 21:28:39 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Histo Tech Mananger Supervisor and Bench Tech's needed for immediate openings Message-ID: Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisor, Histo Managers and Histo Bench Techs. These are fulltime permanent positions. Right now my Hottest jobs are in Colorado and Oregon who are seeking Lab Managers and Lab Supervisor. I have positions in California and Michigan who are seeking Histotech Supervisors. I also have Histo bench positions in Colorado,California,Pennsylvania,Illinois,New Hampshire,Michigan, Georgia,Florida,South Carolina, and Massachusetts. The client is currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recriuter 1-800-466-9919 ext 223 From brian.dias <@t> mail.utexas.edu Tue Aug 2 22:32:38 2005 From: brian.dias <@t> mail.utexas.edu (Brian Dias) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] HPLC and Protein extraction Message-ID: <5.2.0.9.2.20050802222655.00c2ed48@mail.utexas.edu> Hi Everyone, Have just embarked upon HPLC analysis for 5-HT and 5-HIAA in lizards. Got some HPLC data that looked quite good (5-HT and 5-HIAA levels in fmol being different in pharmacologically manipulated animals). But when i normalized the data to amount of protein in the pellet, my results vanished due to the differences in protein levels in my samples (which were all over the place). 1.I figure that if i take the same number of punches per region per animal, my protein levels should not vary drastically within a region. Is that a wise assumption to make? 2. While performing the HPLC, i suspend my tissue punches in perchloric acid and spin it down. Does anyone know of a protocol i might look into to extract protein from this pellet. I'm presently using Sigma's Mammalian Cell Lysis kit and then using a protein assay by Pierce Biotech? Any input will be appreciated. brian ________________________________________________________________________________________ Brian George Dias Graduate Student (Crews lab) Office Phone #: 512-475-6738 The University of Texas at Austin E-mail: brian.dias@mail.utexas.edu Institute for Neuroscience Website: https://webspace.utexas.edu/bgd85/Webpage/BrianD.htm Patterson 56 1 University Station Stop C0930 Austin, TX 78712 ________________________________________________________________________________________ From jkiernan <@t> uwo.ca Wed Aug 3 00:00:29 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Shrinkage of frozen fixed brain tissue mounted on slides References: <20050802231919.58798.qmail@web80305.mail.yahoo.com> Message-ID: <42F04F6D.58BD5FD8@uwo.ca> Your explanation of the measurements is rather hazy, but a 75% linear shrinkage of fixed tissue from water to stained, dehydrated, cleared and mounted is reasonable. For brain tisue this was all reviewed long ago in "Methods in Brain Research", ed. P. B. Bradley; London: John Wiley, in the 1970s. The book is probably in your library. There is a large body of literature, dating back more than a century, about shrinkage. Bear in mind that a section stuck to a slide shrinks less in the plane of the glass than in its thickness when water is removed by any method (alcohol, air-drying etc). John Kiernan Anatomy, UWO London, Canada ______________________________ CM Bush wrote: > > Dear Histonet, > > I have been cutting frozen fixed human brain tissue, using a freezing stage on a rotary microtome. The tissue has been very well fixed in 10%NBF, and cryoprotected with sucrose. > > I cut the sections at 50 um, floated the sections in PBS, mounted them on slides, and allowed them to dry for 48hrs and then Nissl stained. > > Everything looks nice except that the sections have shrunk down to 20um as measured using a stereology set up. This is very sad. > > I guess we will just have to cut thicker sections... > > My question is, does this sound like the amount of shrinkage to expect with frozen fixed brain tissue mounted onto slides? It makes sense that there would be some change due to expansion of freezing water, and then drying on the slide but greater than 50% change? > > Thank you in advance, for your help. > > CM Bush _____ From louise.renton <@t> gmail.com Wed Aug 3 02:29:27 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] slide disposal In-Reply-To: <000301c5979d$3b7721d0$3601a8c0@brownpathology.net> References: <000301c5979d$3b7721d0$3601a8c0@brownpathology.net> Message-ID: Bonnie, I don't klnow if this would be applicable to the USA, but I disposed of slides via a glass recycling works - however they treat the glass for the recycling process renders any biohazard minimal if not non existant. However, not all companies will deal with this type of glass - check first. On 8/2/05, Bonnie Whitaker wrote: > Hi All, > > Can some of you give me ideas of how you handle old slide disposal? > Particularly those of you in smaller, private labs. I have about 20 years > worth of slides that I need to dispose of. If I have to send them with my > biohazardous waste, I can, but it seems like an unnecessary expense. > Currently that is what the procedure says that we do, but is it really > necessary? Maybe someone out there would like to build a glass house : ) > > Bonnie Whitaker > Lab Manager > Brown & Associates Medical Laboratories > 8076 El Rio > Houston, Texas 77054 > 713-741-6677 > > > _______________________________________________ > 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 "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere From Tanni.Ahmed <@t> intervet.com Wed Aug 3 02:48:55 2005 From: Tanni.Ahmed <@t> intervet.com (Ahmed, T (Tanni)) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Does anyone have a spare tissue processor that is getting in the way? Message-ID: <09E7875E806DFB4BBECBADBF966BDBB6D75BAB@mksn79.d50.intra> Peter, We have a old 2L carousel style tissue processor (Shandon) in the archive area which is not in use, but is in good working condition - has been serviced and maintained in recent years. If you are interested, please let me know... Regards, Tanni Tanni S Ahmed Scientific Officer - Histopathology, R&D Intervet UK Ltd. Walton Manor, Walton, Milton Keynes, Buckinghamshire, MK7 7AJ, UK. Tel. +44(0)1908 685552/685543 Fax +44(0)1908 685614 E-mail: tanni.ahmed@intervet.com Message: 10 Date: Tue, 02 Aug 2005 12:01:29 +0100 From: "Peter Bannister" peter_bannister@hotmail.co.uk Subject: [Histonet] Does anyone have a spare tissue processor that is getting in the way? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hello again Histonetters, I realise that this is a slightly cheeky message, but I need to ask if any of you (particularly those based in the UK) have an old carousel style tissue processor taking up valuable storage space in your lab? We are currently setting up a new research lab here in london that will have a relatively low specimen throughput and a rather limited equipment budget. We would therefore be eternally grateful if you would allow us to help you dispose of your old kit!We will of course collect and I will personally chip in towards your christmas party.Yes it's cheeky - but 'if you don't ask, you don't get!' If you can help, please contact me directly. Many thanks for your time. Peter Bannister -------------------------------------- This message, including attachments, is confidential and may be privileged. If you are not an intended recipient, please notify the sender then delete and destroy the original message and all copies. You should not copy, forward and/or disclose this message, in whole or in part, without permission of the sender. -------------------------------------- From Kemlo.Rogerson <@t> elht.nhs.uk Wed Aug 3 01:25:46 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Whole Slide Imaging Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4C4@elht-exch1.xelht.nhs.uk> We have a couple of Coolpics' that I assume are capable of whole slide imaging. But we are in the UK are in the NHS so I don't think we can charge or do it FOC. If you let me now what you are looking at then I'll try our Coolpics out to see if they can do it; you then have some info if you want to buy one. They are about ?35,000 I think so I guess that's $35,000 as we in the UK tend to have to pay ?1 for $1 and I don't know why; History Channel told me it was about 'lend lease' and some unfair bargaining after the 2nd World War by the Americans; is that true? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ben Sent: 02 August 2005 17:04 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Whole Slide Imaging Hello All, I'm interested in whole slide imaging for some research work. I was hoping that anyone who has worked with it could give me their opinion on the equipment they use, the important features (by the way I'm looking to count nuclei), or suggest a good place to get this done for a decent price. If this message is against the rules of histonet, I apologize, but if not I appreciate your help. Ben Renquist UC Davis _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From klapka <@t> uni-duesseldorf.de Wed Aug 3 04:13:33 2005 From: klapka <@t> uni-duesseldorf.de (klapka) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Procollagen IV antibody Message-ID: <42F08ABD.6020706@uni-duesseldorf.de> Hello all, I am looking for an antibody against procollagen type IV. Maybe you know whether I can also use another procollagen antibody? I would like to use it for rat tissue, but if you know anything for human tissue as well, I would like to test the cross reactivity. Thank you , bye, Nicole From Stephen.Eyres <@t> sanofi-aventis.com Wed Aug 3 05:56:33 2005 From: Stephen.Eyres <@t> sanofi-aventis.com (Stephen.Eyres@sanofi-aventis.com) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] Effectiveness of Leica gravity Laser microdissection collection system Message-ID: Hi All, We are in the process of purchasing an LMD. I have picked up a few concerns about the effectiveness of the method used to collect dissected samples. Leica use a gravity system and there appears to be some problems with this, i.e. dissected areas do not fall into the collecting cap cleanly. Can anyone offer their experiences? Many thanks Steve ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- From AFeatherstone <@t> KaleidaHealth.Org Wed Aug 3 06:52:24 2005 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:25:24 2005 Subject: [Histonet] RE: Histonet Digest, Vol 21, Issue 2 Message-ID: <139141F8BAF4A642A945ECC528511AF0012A7841@kalmb02.kaleidahealth.org> The Movat Pentachrome stain will give you your elastic, trichrome and alcian blue. We use it all the time for our temporal biopsies. Annette Featherstone HT/MLT -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, August 02, 2005 13:03 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 21, Issue 2 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Re: Problems of processing brain for paraffin embedding (Gayle Callis) 2. New Lab Design (Breeden, Sara) 3. Results of "New Lab Ideas" Poll (Breeden, Sara) 4. Embedding human cartilage/joint tissue in plastic (Casey, Jane) 5. mouse anti-Her2 (Eva C Andersson) 6. Avoiding shrinkage Re: [Histonet] Embedding human cartilage/joint tissue in plastic (Gayle Callis) 7. Histo tech position available (Cazares, Ruth) 8. Re: staining for iron in microglia (John Kiernan) 9. RE: New lab design (Malam Jacqueline) 10. Does anyone have a spare tissue processor that is getting in the way? (Peter Bannister) 11. ANTI-MYOSIN AND ANTIRENIN (Jose Luis Palazon Fernandez) 12. retina neurons (Jose Luis Palazon Fernandez) 13. cassette labeller (Demarinis, Carolyn) 14. Elastic Trichrome - any good protocol out there not involving microwave??? (GT Hebert) 15. RE: Elastic Trichrome - any good protocol out there no t involving microwave??? (Monfils, Paul) 16. Re: Elastic Trichrome - any good protocol out there not involving microwave??? (Bryan Hewlett) 17. RE: Elastic Trichrome - any good protocol out there no t involving microwave??? (Bonner, Janet) 18. Whole Slide Imaging (Ben) ---------------------------------------------------------------------- Message: 1 Date: Mon, 01 Aug 2005 12:06:41 -0600 From: Gayle Callis Subject: Re: [Histonet] Problems of processing brain for paraffin embedding To: azita parvaneh tafreshi , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050801120504.01b73828@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Are you processing whole brain, or coronal slices? Once you indicate, I will be happy to send our protocols via private email attachement. At 02:30 AM 7/30/2005, you wrote: >Dear Members, >I have problems with paraffin processing of mouse and hamster brains and >spinal cords. I have perfused the animals with PFA 4% and I postfixed them >in the same buffer for 7-8 days and then stored in Alc 70% (up to now). >After sectioning, tissues wrinkle in a way that it is torn after >flattenning (on a slide warmer), therefore the quality is not good at all. > >Can anyone give me hints to overcome the problem. A detailted protocol of >paraffin processing would be great. > >Regard >A.P. Tafreshi Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 2 Date: Mon, 1 Aug 2005 12:33:25 -0600 From: "Breeden, Sara" Subject: [Histonet] New Lab Design To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Just a quick note to thank all of you that sent me your ideas/comments/input on what to put into a new histo lab. These ideas have been duly noted and will be part of My Grand Plan. There were some meaty ideas and some real sparklers! Your help is invaluable -- and if you've thought of anything else since last week, let me know. Thanks again! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Monday, August 01, 2005 11:02 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 21, Issue 1 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Excessive background staining with Millers elastic stain (Lachlan Smith) 2. staining for iron in microglia (Sharon Cooperman) 3. Re: Excessive background staining with Millers elastic stain (Paul Bradbury) 4. RE: New Lab Design (Rogerson Kemlo (ELHT) Pathology) 5. Biotinylating Antibodies (Orr, Rebecca) 6. Re: EGFR (DDittus787@aol.com) 7. (no subject) (Patsy Ruegg) 8. RE: (no subject) (Patsy Ruegg) ---------------------------------------------------------------------- Message: 1 Date: Mon, 1 Aug 2005 09:33:17 +0930 From: "Lachlan Smith" Subject: [Histonet] Excessive background staining with Millers elastic stain To: Message-ID: <000001c5962c$6b43cca0$e46b140a@ITP36161> Content-Type: text/plain; charset="iso-8859-1" Dear Subscribers I am staining cartilage using Millers elastic fibre stain, and while elastic fibres are staining well, there is a lot of blue background staining which makes thresholding for histoquantitation almost impossible. My sections are 30 micron paraffin embedded. I am also oxidising with 0.5% aqueous potassium permanaganate and bleaching with 2% oxalic acid. Any suggestings for reducing this background staining would be greatly appreciated. Kind regards, Lachlan Smith Institute of Medical and Veterinary Science Adelaide, Australia ------------------------------ Message: 2 Date: Sun, 31 Jul 2005 21:39:40 -0400 From: Sharon Cooperman Subject: [Histonet] staining for iron in microglia To: Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Dear Histonetters, I have a semi-scientific question: macrophages in almost all tissues can be stained for iron with Perl's stain to give a blue color - is this also true for microglia? If not, is there any other way to see how much iron is in microglia (DAB enhanced Perl's or some other method)? Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 ------------------------------ Message: 3 Date: Sun, 31 Jul 2005 22:31:43 -0700 From: Paul Bradbury Subject: Re: [Histonet] Excessive background staining with Millers elastic stain To: Lachlan Smith , HistoNet Server Message-ID: <42EDB3BF.3030304@shaw.ca> Content-Type: text/plain; format=flowed; charset=us-ascii Hi Lachlan, I believe the problem you are having is due to the presence of very similar reactive groups in both cartilage and elastic fibres, Elastin (the protein ) is heavily sulphated with an abundance of disulphide groups (hence the yellowish appearance of elastic fibres when viewed unstained or even macroscopically). When you oxidize the sections in potassium permanganate, you are converting the disulphides to sulphates, the sulphates react with Miller's elastin stain. Miller's stain is essentially as modification of aldeyde fuchsin which iis a well known method for demonstrating sulphated groups in mucopolysaccharides, etc. The extracellular matrix of cartilage consists of chondroitin sulphate (plus a few other assorted bits of connective tissue). These groups will react quite strongly with Miller's stain solution. So unfortunately, I think you may well be out of luck as far as improving the specificity of the staining reactions. The same low specificity reactions occur with most other elastin staining methods when they are used on cartilage, the reactive groups present in both locations stain with the dye in use. I cannot think of a method that stains only elastin and not cartilage matrix. Thinner sections may give you a better chance of distinguishing one from the other. Try sections at several thinner settings and see what the outcome is. Maybe there is a thickness at which you can see sufficient enough detail in the elastic fibres without the cartilage obscuring it. Paul Bradbury, Kamloops, BC Canada Lachlan Smith wrote: >Dear Subscribers > >I am staining cartilage using Millers elastic fibre stain, and while >elastic fibres are staining well, there is a lot of blue background >staining which makes thresholding for histoquantitation almost >impossible. My sections are 30 micron paraffin embedded. I am also >oxidising with 0.5% aqueous potassium permanaganate and bleaching with >2% oxalic acid. > >Any suggestings for reducing this background staining would be greatly >appreciated. > >Kind regards, > >Lachlan Smith >Institute of Medical and Veterinary Science Adelaide, Australia > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ Message: 4 Date: Mon, 1 Aug 2005 11:06:44 +0100 From: "Rogerson Kemlo (ELHT) Pathology" Subject: RE: [Histonet] New Lab Design To: "Breeden, Sara" , Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4C0@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" AFOS (or something similar), formalin extraction for grossing. Includes a downdraught table and formalin making up facilities. AFOS formalin extraction cabinets for stored samples in formalin. Be careful over height of benches; some are for sitting down carrying out work, others for cutting sections using a microtome. Nothing worse than a microtome that's too high or too low. You need solvent extraction hoods for your processing machines, a suitable flammable store with antiflash lighting etc., The best lighting to get it that stuff they sell that goes in Offices; it's expensive but gives you a good colour temperature with few shadows. If you can see, stop people asphyxiating on formalin and alcohol, stop the place blowing up and have benches the right height that people can work, nowt else much to do. Oh yes, coffee machine, clean area and a Mars Bar vending machine. Ergo, all done! -----Original Message----- From: Breeden, Sara [mailto:sbreeden@nmda.nmsu.edu] Sent: 29 July 2005 17:27 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] New Lab Design A new histology lab (in a new building) is in the works (yipee!!). I have some definite ideas about what I'll need in my new digs, but I need more opinions! Within the next couple weeks, I will be asked by the architects for my input. This is a State veterinary lab where I do "surgicals", necropsies, standard special stains, with a rapidly expanding IHC workload. My total workload was roughly 10,000 blocks last year and I expect it to grow by 5% each year. I expect to have roughly 500 sq ft to work with in this new space. I work alone for three DVM pathologists. What would you consider absolutely essential in a new histology lab? What would you do differently if you'd had a chance? What kind of: bench space and type of surface, ventilation, lighting, hoods, haz mat storage, supply storage, windows? I only get once chance at this and I want to make it count...I have 8 more years until retirement and I'd rather not kick myself for missing something obvious. Please reply directly to me. Thank you in advance!! I rely on 'netters for great information. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 1 Aug 2005 07:26:53 -0500 From: "Orr, Rebecca" Subject: [Histonet] Biotinylating Antibodies To: Message-ID: Content-Type: text/plain; charset="us-ascii" Vernon, I have been successful with a biotinylation kit from Biocare Medical. It uses a different chemistry than the Vector kits. I think the mopping reagent may be what makes the difference. Their number is 800 799 9499 ask for Hari, he's the technical sales manager. Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 ------------------------------ Message: 6 Date: Mon, 1 Aug 2005 08:46:13 EDT From: DDittus787@aol.com Subject: Re: [Histonet] EGFR To: JosefaNava@texashealth.org, histonet@pathology.swmed.edu Message-ID: <9b.648c607f.301f7395@aol.com> Content-Type: text/plain; charset="US-ASCII" Josie: Are you aware of the clone 31g7 and Allen Gowns ASCO 2005 poster. It showed that 31g7 from Zymed was best in class and selected 10-15% more patients. dana ------------------------------ Message: 7 Date: Mon, 1 Aug 2005 10:21:23 -0600 From: "Patsy Ruegg" Subject: [Histonet] (no subject) To: "'Histonet'" Message-ID: <200508011621.j71GLFpf017202@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Does anyone know where I could get a service manual for an old IEC cryostat? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. ------------------------------ Message: 8 Date: Mon, 1 Aug 2005 10:23:37 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] (no subject) To: , Message-ID: <200508011623.j71GNTpf017949@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" We used picro sirrus stain for collagen fibers and viewed the collagen bundles using polarized light filter for histomorphometry. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of lachlan.smith@imvs.sa.gov.au Sent: Sunday, July 31, 2005 3:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] (no subject) Dear Subscribers I am staining cartilage using Millers elastic fibre stain, and while elastic fibres are staining well, there is a lot of blue background staining which makes thresholding for histoquantitation almost impossible. My sections are 30 micron paraffin embedded. I am also oxidising with 0.5% aqueous potassium permanaganate and bleaching with 2% oxalic acid. Any suggestings for reducing this background staining would be greatly appreciated. Kind regards, Lachlan Smith Institute of Medical and Veterinary Science Adelaide, Australia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 21, Issue 1 *************************************** ------------------------------ Message: 3 Date: Mon, 1 Aug 2005 13:17:21 -0600 From: "Breeden, Sara" Subject: [Histonet] Results of "New Lab Ideas" Poll To: Message-ID: Content-Type: text/plain; charset="US-ASCII" And with many thanks to all of you that had input, herewithin is a list of the thoughts. And, might I add, this is just one reason I think this medium (Histonet) is absolutely necessary!! Lookie what I found out!!... Large SINKS - and more than one; sink in HOOD for specials, IHC, etc.; ergonomically correct SEATING and COUNTER HEIGHT for task; proper VENTILATION and AIR DIRECTION (as in no airflow above microtome/water bath, but proper ventilation for fume removal and room temperature control); BRIGHT WHITE LIGHTING in task areas; VENT (ambient air pressure?) or similar over embedding area/processor (for heat removal); DEDICATED CIRCUITS for essential equipment and BACKUP POWER UNITS for same; D.I. WATER; separate HAZMAT STORAGE and STORAGE for acids and bases (separately); STORAGE for supplies/inventory (larger than you think you'll ever need); EYEWASH/SHOWER; TEXTURED FLOOR; 30" deep COUNTERS; rounded COUNTERTOP edges; COUNTERTOPS that don't stain or corrode (stainless steel not recommended); no GLASS-INSERT DOORS (visible clutter); overhead VENTS w/snorkels; and appropriate WORKFLOW planning. Of course, this prompted a few "needs" for my particular space, which I will list just for informational purposes. But it may jog a thought for someone in a similar position: HAZMAT storage out of work area or in an area that's near the need (i.e., processor); trash cans out of walkway (!); area for slide return (post-pathologist); locking cabinets and drawers (for blades, knives, Secret Histology Stuff, etc.); and under-cabinet lighting. I'm sure the list will grow. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX ------------------------------ Message: 4 Date: Mon, 1 Aug 2005 15:22:58 -0400 From: "Casey, Jane" Subject: [Histonet] Embedding human cartilage/joint tissue in plastic To: Message-ID: <471F15E271937A4DA89AC12211DDAD980266E9E3@MAIL2.AD.Brown.Edu> Content-Type: text/plain; charset="iso-8859-1" We are trying to use histology to validate our method of measuring cartilage thickness on wrist bones using ?CT scans. Initially we tried fixing a trapezoid fragment in formalin, de-mineralizing it, and embedding it in paraffin before slicing it into 5 ?m sections. This method led to non-uniform shrinkage of the tissue and clearly visible artifact. We would like to try a method, perhaps embedment in plastic, which will yield a more accurate measure of cartilage thickness. Does anyone have any suggestions? Thank you for your suggestions. Jane Casey Department of Orthopaedics Brown Medical School / Rhode Island Hospital Providence, RI 02912 Jane_Casey@Brown.edu ------------------------------ Message: 5 Date: Mon, 01 Aug 2005 16:25:34 -0400 From: Eva C Andersson Subject: [Histonet] mouse anti-Her2 To: histonet@lists.utsouthwestern.edu Message-ID: <42EE853E.2010103@georgetown.edu> Content-Type: text/plain; charset=us-ascii; format=flowed Hello Everyone, I just wanted to ask about a mouse anti-Her2 clone CB11 antibody that I have bought from Zymed. It says the optimal dilutions are to be determined by each lab but I just want to know if someone could tell me which dilution they use. Also do you use an antigen retrieval method and which detection method do you use? Thank you Eva Georgetown University ------------------------------ Message: 6 Date: Mon, 01 Aug 2005 14:28:34 -0600 From: Gayle Callis Subject: Avoiding shrinkage Re: [Histonet] Embedding human cartilage/joint tissue in plastic To: "Casey, Jane" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050801141209.01b3ac10@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed A clever, simple use of low voltage xray machines i.e FAXITRON can help you out. We did specimen radiographs with a FAXITRON, and underexposed the sample to enhance soft tissues i.e tumors and cartilage. These will be like "contact" photographs, showing no shrinkage of the soft tissue components. After fixation DO not decalcify, radiograph the sample then decalcify (by the way, xray is the most sensitive decalcification endpoint determination) and do another radiograph at the same initial exposure. You can do thicker slabs even 5 mm thick but make sure you are consistent with thickness of slab, a good bone saw will do this - Buehler Isomet or the MarMed saw, a lovely device. The bones can be NBF fixed but you avoid all dehydration/clearing and heat of paraffin which will cause shrinkage. If you have to know exact edge of cartilage, a radioopaque substance can be painted on surface of cartilage to define that sharp edge. There is a publication where a group tested the amount of shrinkage you get with both paraffin and plastic processing, results were interesting in that BOTH methods caused as much as 20% shrinkage, possibly more of the hard tissue, so you will not avoid shrinkage with plastic embedding methods involving alcohols or any other solvent to remove water. Since cartilage has a high water content and by doing specimen radiography, you can avoid alcohols, clearing agents and heat of paraffin that cause shrinkage by water removal. At 01:22 PM 8/1/2005, you wrote: > > >We are trying to use histology to validate our method of measuring >cartilage thickness on wrist bones using ?CT scans. Initially we tried >fixing a trapezoid fragment in formalin, de-mineralizing it, and embedding >it in paraffin before slicing it into 5 ?m sections. This method led to >non-uniform shrinkage of the tissue and clearly visible artifact. We would >like to try a method, perhaps embedment in plastic, which will yield a >more accurate measure of cartilage thickness. Does anyone have any >suggestions? > >Thank you for your suggestions. > > >Jane Casey >Department of Orthopaedics >Brown Medical School / Rhode Island Hospital >Providence, RI 02912 >Jane_Casey@Brown.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) ------------------------------ Message: 7 Date: Mon, 1 Aug 2005 16:09:48 -0500 From: "Cazares, Ruth" Subject: [Histonet] Histo tech position available To: Message-ID: <913FAC2B773C19488E26AE6572180FA5029A992C@exch01.schosp.org> Content-Type: text/plain; charset="us-ascii" Hi all, We have a position open for a histo tech at Swedish Covenant Hospital in Chicago. The position is alternate Saturdays (8 hour day) and fill in for vacations if possible. There is also opportunity to pick up extra hours during the week on evenings but this is optional. There will also be a full time position open in December when one of my FT techs retires. Interested techs (certified) can call or fax their resume; Ruth Cazares Department of Pathology Phone: 773 878-8200 ext-5190 Fax: 773 271-1501 (to my attention) *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. ------------------------------ Message: 8 Date: Tue, 02 Aug 2005 00:16:09 -0400 From: John Kiernan Subject: Re: [Histonet] staining for iron in microglia To: Sharon Cooperman , histonet@lists.utsouthwestern.edu Message-ID: <42EEF389.A65BF4AC@uwo.ca> Content-Type: text/plain; charset=us-ascii Sharon, Your question is 100% scientific, not semi-scientific! You concisely provide an introduction, expand it into a testable hypothesis, and ask if the test has been done. The answer is "Yes, but ..." Iron is seen in phagocytic cells in various pathological conditions of the brain. Its presence in the frontal cortex (seen with the regular Perls method) is a classical feature of advanced syphilis (general paralysis of the insane). There are some cells in the normal rat's hypothalamus that have a variety of histochemical peculiarities including being Perls-positive, but these cells are astrocytes (Young & McKenzie, 2004). The DAB-enhanced Perls method shows non-haem iron in microglia, oligodendroglia and neurons. Research in this field, from several labs, was published in the late 1980s and early 1990s, about 10 years after the publication of the DAB amplification procedure by Nguyen-Legros et al in 1980. A few references are given at the end of this message. They are ones that I've seen and made notes on; there will be many more, findable by way of PubMed, Web of Science or Scopus. At NIH all those resources and more will be available to you. Another iron-related approach to microglia has been immunostaining for ferritin (Kaneko et al 1989). There are some lectins with affinity for microglia that can provide impressive pictures when correctly applied. For the old and bold there are the original silver stains, developed by Del Rio Hortega and perfected by Penfield and Cone in the 1920s. Summary. Sensitive methods for iron do not provide specific staining of resident, activated or immigrant microglial cells. The classical Perls method can detect microglia that have collected iron in sites of injury or disease. Iron is also present in macroglia and neurons, and is stainable in these cells if the sensitivity of the Perls prussian blue method is enhanced. References. Gerber MR, Connor JR (1989) Do oligodendrocytes mediate iron regulation in the human brain? Ann. Neurol. 26: 95-98. Kaneko Y, Kitamoto T, Tateishi J, Yamaguchi K (1989) Ferritin immunohistochemistry as a marker for microglia. Acta Neuropathol. (Berl.) 79: 129-136. Morris CM, Candy JM, Oakley AE, Bloxham CA, Edwardson JA (1992) Histochemical distribution of non-haem iron in the human brain. Acta Anat. 144: 235-257. Connor JR, Pavlick G, Karli D, Menzies SL, Palmer C (1995) A histochemical study of iron-positive cells in the developing rat brain. J. Comp. Neurol. 355: 111-123. Palmer C, Menzies SL, Roberts RL, Pavlick G, Connor JR (1999) Changes in iron histochemistry after hypoxic-ischemic brain injury in the neonatal rat. J. Neurosci. Res. 56: 60-71. Young JK, McKenzie JC (2004) GLUT2 immunoreactivity in Gomori-positive astrocytes of the hypothalamus. J. Histochem. Cytochem. 52: 1519-1524. John Kiernan Anatomy, UWO London, Canada --------------------------------------- Sharon Cooperman wrote: > > Dear Histonetters, > > I have a semi-scientific question: macrophages in almost all tissues > can be stained for iron with Perl's stain to give a blue color - is > this also true for microglia? If not, is there any other way to see > how much iron is in microglia (DAB enhanced Perl's or some other > method)? > > Thanks, > Sharon > -- > Sharon Cooperman > NIH, NICHD, CBMB 301.435-8417 > Building 18T, room 101 301.402-0078 fax > Bethesda, MD 20892 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Tue, 2 Aug 2005 08:41:07 +0100 From: Malam Jacqueline Subject: [Histonet] RE: New lab design To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain One thing you need to over-estimate on is block and slide storage, and if you are not on a ground floor, they weigh a ton so you'll need some additional support system. Cheers Jacqui Malam DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. ------------------------------ Message: 10 Date: Tue, 02 Aug 2005 12:01:29 +0100 From: "Peter Bannister" Subject: [Histonet] Does anyone have a spare tissue processor that is getting in the way? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hello again Histonetters, I realise that this is a slightly cheeky message, but I need to ask if any of you (particularly those based in the UK) have an old carousel style tissue processor taking up valuable storage space in your lab? We are currently setting up a new research lab here in london that will have a relatively low specimen throughput and a rather limited equipment budget. We would therefore be eternally grateful if you would allow us to help you dispose of your old kit! We will of course collect and I will personally chip in towards your christmas party. Yes it's cheeky - but 'if you don't ask, you don't get!' If you can help, please contact me directly. Many thanks for your time. Peter Bannister _________________________________________________________________ It's fast, it's easy and it's free. Get MSN Messenger 7.0 today! http://messenger.msn.co.uk ------------------------------ Message: 11 Date: Tue, 2 Aug 2005 13:36:04 +0200 (CEST) From: Jose Luis Palazon Fernandez Subject: [Histonet] ANTI-MYOSIN AND ANTIRENIN To: Histonet@lists.utsouthwestern.edu Message-ID: <20050802113604.74D8A89C1F5@perceval.uca.es> Content-Type: text/plain; charset="iso-8859-1" Dear List-members I am finishing my doctoral thesis and would like to demonstrate renin in an aglomerular teleost kidney, smooth muscle in the iris, and muscle in the bulbus arteriosus. I know that this can be done inmunohistologically but my lab does not work in inmunohistochemistry so I do not have the antibodies. I would like to know if there is someone of our list here in Spain (hospital, University) that can do this kind of inmunos and the posibility of doing my samples with them or maybe sending the samples and paying for the inmunos. thanks in advance Jos? Luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es ------------------------------ Message: 12 Date: Tue, 2 Aug 2005 13:37:43 +0200 (CEST) From: Jose Luis Palazon Fernandez Subject: [Histonet] retina neurons To: Histonet@lists.utsouthwestern.edu Message-ID: <20050802113743.3F3E989C5A1@perceval.uca.es> Content-Type: text/plain; charset="iso-8859-1" Dear list fellows is there any simple stain that I can use to differentiate between amacrine, horizontal and bipolar cells in the retina. I have tried using H&E but as I have not experience in this kind of tissue I dont know how to differentiate between them, mainly between amacrine and bipolar cells. thanks in advance jose luis Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de Ciencias Marinas de Andalucia Puerto Real, C?diz, Espa?a. email: jluis.palazon@icman.csic.es ------------------------------ Message: 13 Date: Tue, 2 Aug 2005 07:43:35 -0400 From: "Demarinis, Carolyn" Subject: [Histonet] cassette labeller To: "HISTONET \(E-mail\)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" We are interested in obtaining a cassette labeller. Does anyone have any recommendations? We are a Meditech hospital. **************************************************************************** *********** CONFIDENTIALITY NOTICE: This e-mail communication and any attachmentsmay 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. If you have received this communication in error, please notify Saratoga Hospital immediately by e-mail at privacy@saratogacare.org and destroy all copies of this communication and any attachments. ------------------------------ Message: 14 Date: Tue, 2 Aug 2005 06:05:22 -0700 (PDT) From: GT Hebert Subject: [Histonet] Elastic Trichrome - any good protocol out there not involving microwave??? To: histonet@lists.utsouthwestern.edu Message-ID: <20050802130522.4981.qmail@web31710.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello all, I am in need of a good protocol for a combination elastic and trichrome stain that doesn't involve microwave. Thanks. Gustave Hebert Scientist II Wyeth Research Cambridge MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 15 Date: Tue, 2 Aug 2005 09:52:22 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] Elastic Trichrome - any good protocol out there no t involving microwave??? To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171759D@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="ISO-8859-1" I often combine the Verhoeff Elastin Stain with the Gomori Trichrome. I do the Bouin's treatment required for the trichrome procedure first. Then I do the complete Verhoeff stain, eliminatiing the final Van Gieson counterstain. Then I apply the one-step Gomori Trichrome. It is quite simple and gives excellent results. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of GT > Hebert > Sent: Tuesday, August 2, 2005 6:05 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Elastic Trichrome - any good protocol out there > not involving microwave??? > > Hello all, > > I am in need of a good protocol for a combination elastic and trichrome > stain that doesn't involve microwave. Thanks. > > Gustave Hebert > Scientist II > Wyeth Research > Cambridge MA > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 16 Date: Tue, 2 Aug 2005 10:20:58 -0400 From: "Bryan Hewlett" Subject: Re: [Histonet] Elastic Trichrome - any good protocol out there not involving microwave??? To: "Monfils, Paul" , Message-ID: <01a301c5976d$67bdd250$6400a8c0@mainbox> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Paul, The Van Gieson counterstain IS a trichrome! It also has the advantage of not requiring the Bouin pretreatment. An excellent alternative to Verhoff, is the Miller elastic stain which also incorporates the Van Gieson. Bryan ----- Original Message ----- From: "Monfils, Paul" To: Sent: Tuesday, August 02, 2005 9:52 AM Subject: RE: [Histonet] Elastic Trichrome - any good protocol out there not involving microwave??? >I often combine the Verhoeff Elastin Stain with the Gomori Trichrome. I do > the Bouin's treatment required for the trichrome procedure first. Then I > do > the complete Verhoeff stain, eliminatiing the final Van Gieson > counterstain. > Then I apply the one-step Gomori Trichrome. It is quite simple and gives > excellent results. > > Paul M. > >> ---------- >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of GT >> Hebert >> Sent: Tuesday, August 2, 2005 6:05 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Elastic Trichrome - any good protocol out there >> not involving microwave??? >> >> Hello all, >> >> I am in need of a good protocol for a combination elastic and trichrome >> stain that doesn't involve microwave. Thanks. >> >> Gustave Hebert >> Scientist II >> Wyeth Research >> Cambridge MA >> >> __________________________________________________ >> Do You Yahoo!? >> Tired of spam? Yahoo! Mail has the best spam protection around >> http://mail.yahoo.com >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Tue, 2 Aug 2005 10:31:15 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] Elastic Trichrome - any good protocol out there no t involving microwave??? To: "'Bryan Hewlett '" , "'histonet-bounces@lists.utsouthwestern.edu '" , "'Monfils, Paul '" , "'histonet@lists.utsouthwestern.edu '" Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB434C@fh2k093.fhmis.net> Content-Type: text/plain; charset=iso-8859-1 Generally, we find that the steps are so short anyway that we don't use the microwave on the masson's trichrome-except for the Bouin's Solution and even then we try not to because the solution becomes compromised. ---Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Monfils, Paul; histonet@lists.utsouthwestern.edu Sent: 8/2/2005 10:20 AM Subject: Re: [Histonet] Elastic Trichrome - any good protocol out there not involving microwave??? Paul, The Van Gieson counterstain IS a trichrome! It also has the advantage of not requiring the Bouin pretreatment. An excellent alternative to Verhoff, is the Miller elastic stain which also incorporates the Van Gieson. Bryan ----- Original Message ----- From: "Monfils, Paul" To: Sent: Tuesday, August 02, 2005 9:52 AM Subject: RE: [Histonet] Elastic Trichrome - any good protocol out there not involving microwave??? >I often combine the Verhoeff Elastin Stain with the Gomori Trichrome. I do > the Bouin's treatment required for the trichrome procedure first. Then I > do > the complete Verhoeff stain, eliminatiing the final Van Gieson > counterstain. > Then I apply the one-step Gomori Trichrome. It is quite simple and gives > excellent results. > > Paul M. > >> ---------- >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of GT >> Hebert >> Sent: Tuesday, August 2, 2005 6:05 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Elastic Trichrome - any good protocol out there >> not involving microwave??? >> >> Hello all, >> >> I am in need of a good protocol for a combination elastic and trichrome >> stain that doesn't involve microwave. Thanks. >> >> Gustave Hebert >> Scientist II >> Wyeth Research >> Cambridge MA >> >> __________________________________________________ >> Do You Yahoo!? >> Tired of spam? Yahoo! Mail has the best spam protection around >> http://mail.yahoo.com >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Tue, 2 Aug 2005 09:03:42 -0700 From: "Ben" Subject: [Histonet] Whole Slide Imaging To: Message-ID: <200508021603.j72G3gUj002726@pop19.ucdavis.edu> Content-Type: text/plain; charset="us-ascii" Hello All, I'm interested in whole slide imaging for some research work. I was hoping that anyone who has worked with it could give me their opinion on the equipment they use, the important features (by the way I'm looking to count nuclei), or suggest a good place to get this done for a decent price. If this message is against the rules of histonet, I apologize, but if not I appreciate your help. Ben Renquist UC Davis ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 21, Issue 2 *************************************** CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From bruyntjes <@t> voeding.tno.nl Wed Aug 3 08:39:08 2005 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] fixation of fresh frozen section Message-ID: <3B070848E7C2204F9DEB8BCFD7677280055CE513@ntexch1.voeding.tno.nl> Hi all When I do immunocytochemistry on fresh frozen section I fix most, if not all slides with acetone. Sometimes I use cold methanol because the datasheet tells me to do so. Is anyone of you who prefer other fixatives? Joost Bruijntjes TNO Quality of Life Zeist Holland This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From contact <@t> excaliburpathology.com Wed Aug 3 10:04:16 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Tiny 7x7mm base embedding molds Message-ID: <20050803150416.3091.qmail@web50308.mail.yahoo.com> Hello there, I am in search of the tiny 7x7mm base embedding molds. They are the perfect size for my mouse eyes. I can use either the metal or plastic disposable ones. I have an extra, brand new box of the disposable ones, measuring 15x15mm, I will trade. Paula Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From mclarke <@t> allsaintshealthcare.org Wed Aug 3 10:20:03 2005 From: mclarke <@t> allsaintshealthcare.org (Clarke, Mary) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Disposal of glass slides Message-ID: <6FE2A453DA437F4D96FAAF363F20ABB20FC86E@WFEXBE04.wfsi.priv> My safety officer has told us that we can dispose of glass slides in the regular trash, because the tissue is fixed and considered no longer hazardous. Terri Clarke Histology Supervisor All Saints Laboratory 3801 Spring Street Racine, Wi 53405 mclarke@allsaintshealthcare.org 262-687-6609 Privileged/Confidential information may be contained in this message. The information contained in this message is intended only for the use of the recipient(s) named above and their co-workers who are working on the same matter. The recipient of this information is prohibited from disclosing the information to any other party unless this disclosure has been authorized in advance. If you are not intended recipient of this message or any agent responsible for delivery of the message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or action taken in reliance on the contents of this message is strictly prohibited. You should immediately destroy this message and kindly notify the sender by reply E-Mail. Please advise immediately if you or your employer does not consent to Internet E-Mail for messages of this kind. Opinions, conclusions and other information in this message that do not relate to the official business of the firm shall be understood as neither given nor endorsed by it. From gcallis <@t> montana.edu Wed Aug 3 11:22:34 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Re: fixation of fresh frozen section In-Reply-To: <3B070848E7C2204F9DEB8BCFD7677280055CE513@ntexch1.voeding.t no.nl> References: <3B070848E7C2204F9DEB8BCFD7677280055CE513@ntexch1.voeding.tno.nl> Message-ID: <6.0.0.22.1.20050803102107.01b41840@gemini.msu.montana.edu> Joost, What are you staining for, i.e. what antigens? That determines what fixative we choose to use. At 07:39 AM 8/3/2005, you wrote: >Hi all > > > >When I do immunocytochemistry on fresh frozen section I fix most, if not >all slides with acetone. Sometimes I use cold methanol because the >datasheet tells me to do so. Is anyone of you who prefer other >fixatives? >Joost Bruijntjes >TNO Quality of Life >Zeist >Holland > > 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 Wed Aug 3 11:25:09 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Whole Slide Imaging References: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4C4@elht-exch1.xelht.nhs.uk> Message-ID: <001a01c59847$eaa04880$7e034246@yourlk4rlmsu> ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" "we in the UK tend to have to pay ?1 for $1 and I don't know why; History Channel told me it was about 'lend lease' and some unfair bargaining after the 2nd World War by the Americans; is that true?" When I was growing up in North London in the 50's and early 60's, the excuse for high prices was "we still have the debt from the first world war". I'm sure that in negotiations after WW2, the US bargained for its own national interest, as did the US (and Canada, and France, and .......). However, I don't think British economic policy has much to do with what happened during negotiations over who won the war 60 years ago, pre-EU. I suspect the real reason for British pound for dollar costs is commercial greed and taxes. Commercial greed is otherwise known as pricing according to "what the market will bear", or "gouge the buggers for every penny they've got". As for taxes, well... bureaucracy costs and the UK has one hell of a bureaucracy from what I recall before I emigrated. Bryan Llewellyn From gcallis <@t> montana.edu Wed Aug 3 11:30:38 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Re: Disposal of glass slides In-Reply-To: <6FE2A453DA437F4D96FAAF363F20ABB20FC86E@WFEXBE04.wfsi.priv> References: <6FE2A453DA437F4D96FAAF363F20ABB20FC86E@WFEXBE04.wfsi.priv> Message-ID: <6.0.0.22.1.20050803102857.01b6af50@gemini.msu.montana.edu> If you can dispose of slides in trash, how about large broken glass diposal box - safer for janitors and other personnel. At 09:20 AM 8/3/2005, you wrote: >My safety officer has told us that we can dispose of glass slides in the >regular trash, because the tissue is fixed and considered no longer >hazardous. > >Terri Clarke 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 David.Edmondson <@t> christie-tr.nwest.nhs.uk Wed Aug 3 11:36:46 2005 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] fixation of fresh frozen section Message-ID: Hi I recently heard a presentation where they spoke of Periodate-Lysine-Paraformaldehyde in the context of Frozen sections. The coagulant fixatives do not alter the proteins chemistry so on might suggest no loss of antigenicity, the "Gold Standard". But the morphology of PLP is much better. It is fifteen and more years since I used it myself and maybe a search would be a quicker way to find the concentrations. I ahve an email contact if you need to ask him. David Christie Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bruijntjes, J.P. Sent: 03 August 2005 14:39 To: histonet@pathology.swmed.edu Subject: [Histonet] fixation of fresh frozen section Hi all When I do immunocytochemistry on fresh frozen section I fix most, if not all slides with acetone. Sometimes I use cold methanol because the datasheet tells me to do so. Is anyone of you who prefer other fixatives? Joost Bruijntjes TNO Quality of Life Zeist Holland This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From contact <@t> excaliburpathology.com Wed Aug 3 11:40:46 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Tiny 7x7 molds Message-ID: <20050803164046.36329.qmail@web50308.mail.yahoo.com> Wow, thanks to all that offered to trade embedding molds. I am now getting out the brown wrapping paper to send you yours. Paula Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From sharon.osborn <@t> dnax.org Wed Aug 3 11:56:47 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Slide Labeler Message-ID: <29B25753F6B1D51196110002A589D444023980F7@PALMSG30.us.schp.com> I have used the TBS slide etch labeler and cassette labeler for over 10 years. At the time, they were one of the few on the market that could handle the volume of the laboratories. Now, there is a really good system available in the Leica printers. The technology is trouble free in comparison to the etching systems available. You have greater choices of what information to place onto the cassettes and slides. I suggest you demo any that you are interested in before making a final decision; or, go visit laboratories that have the automated labelers and ask lots of questions while watching it work. The Leica rep brings prospective clients to our lab to view the Leica printers and to talk freely with us. What problems we have, the service and sales reps both work quickly to solve. It is so much easier and nicer than with the previous labeler. sharon osborn DNAX, SP BioPharma Palo Alto, CA Date: Tue, 2 Aug 2005 07:43:35 -0400 From: "Demarinis, Carolyn" Subject: [Histonet] cassette labeller To: "HISTONET \(E-mail\)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" We are interested in obtaining a cassette labeller. Does anyone have any recommendations? We are a Meditech hospital. ********************************************************************* 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 liz <@t> premierlab.com Wed Aug 3 13:42:30 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] cytospin preps and plus slides Message-ID: <000001c5985b$1c3023d0$a7d48a80@AMY> Hello Everyone I'm working on a project for a client who is trying to isolate prostate epithelial cells. I'm getting the cells submitted to me fixed in 10% NBF in eppendorf tubes. I'm using a cytospin for 5 minutes at 1000 rpm and plus slides. The problem is that I'm not getting the correct yield of cells. They check the number of cells in the prep prior to me receiving them. They are submitting numbers of cells in the thousands, but when I prepare the cytospin slides, I'm only getting cells numbering in the hundreds and sometimes less. Does anyone have any suggestions? Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 9msm8 <@t> qlink.queensu.ca Wed Aug 3 13:53:35 2005 From: 9msm8 <@t> qlink.queensu.ca (9msm8@qlink.queensu.ca) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Casp-3 advice Message-ID: <51618.192.75.165.28.1123095215.squirrel@qlink.queensu.ca> Hello Histonet. I am a student in a lab where histologists are rare to non-existant. I am working with 5 micrometer thick mouse hippocampal parrafin embedded sections (I used paraformaldehyde) and I want to do an immunohistochemical stain for casp-3. I am using the marker as a detection for relatively early apoptosis. The QUESTION I am asking is: Should I be using this method with a fluorescence signal or a horshradish peroxidase counterstain? Given that I probably will want my analysis to be more of a qualitative descritpion. What should I be aware of before making my decision? What are the advantages and disadvantages? Any help will be greatly appreciated. From LewisS <@t> ccri.net Wed Aug 3 14:32:00 2005 From: LewisS <@t> ccri.net (Lewis, Sarah) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] fixation of fresh frozen section Message-ID: <714B9F12B4E18C4C843B66E8E190F2AD447562@res2k3ms01.CRII.ORG> I would love more information on this subject. Thanks in advance!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net -----Original Message----- From: Edmondson David (RBV) NHS Christie Tr [mailto:David.Edmondson@christie-tr.nwest.nhs.uk] Sent: Wednesday, August 03, 2005 12:37 PM To: Bruijntjes, J.P. Cc: Histonet (E-mail 2) Subject: RE: [Histonet] fixation of fresh frozen section Hi I recently heard a presentation where they spoke of Periodate-Lysine-Paraformaldehyde in the context of Frozen sections. The coagulant fixatives do not alter the proteins chemistry so on might suggest no loss of antigenicity, the "Gold Standard". But the morphology of PLP is much better. It is fifteen and more years since I used it myself and maybe a search would be a quicker way to find the concentrations. I ahve an email contact if you need to ask him. David Christie Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bruijntjes, J.P. Sent: 03 August 2005 14:39 To: histonet@pathology.swmed.edu Subject: [Histonet] fixation of fresh frozen section Hi all When I do immunocytochemistry on fresh frozen section I fix most, if not all slides with acetone. Sometimes I use cold methanol because the datasheet tells me to do so. Is anyone of you who prefer other fixatives? Joost Bruijntjes TNO Quality of Life Zeist Holland This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Aug 3 15:55:57 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] PLP fixation info RE: fixation of fresh frozen section In-Reply-To: <714B9F12B4E18C4C843B66E8E190F2AD447562@res2k3ms01.CRII.ORG > References: <714B9F12B4E18C4C843B66E8E190F2AD447562@res2k3ms01.CRII.ORG> Message-ID: <6.0.0.22.1.20050803134024.01b5c108@gemini.msu.montana.edu> Sarah, This was a previous message on Histonet concerning PLP fixation. We have great success with this for mouse and hamster perfusion fixation for a prion study then paraffin processing of tissues. The recipe for PLP has been put on Histonet previously, going to Histonet archives should access fixative recipe. I haven't had success with frozen section fixation, although some have done so, my impatience in taking time to work with it to do immunofluorescence work. Bob Chiovette wrote: 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 Good luck Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From cindipqr <@t> wowway.com Wed Aug 3 19:17:07 2005 From: cindipqr <@t> wowway.com (cindipqr@wowway.com) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Re: Disposal of glass slides In-Reply-To: <6.0.0.22.1.20050803102857.01b6af50@gemini.msu.montana.edu> References: <6FE2A453DA437F4D96FAAF363F20ABB20FC86E@WFEXBE04.wfsi.priv> <6.0.0.22.1.20050803102857.01b6af50@gemini.msu.montana.edu> Message-ID: <20050803231204.M24570@wowway.com> Our facility likes them in smaller boxes, taped shut and labeled as glass slides for disposal. This way it's safe and not as heavy and bulky as the regular glass disposal boxes. Cindi HFH Detroit Mi ---------- Original Message ----------- From: Gayle Callis To: "Clarke, Mary" , Histonet@lists.utsouthwestern.edu Sent: Wed, 03 Aug 2005 10:30:38 -0600 Subject: [Histonet] Re: Disposal of glass slides > If you can dispose of slides in trash, how about large broken glass > diposal box - safer for janitors and other personnel. > > At 09:20 AM 8/3/2005, you wrote: > >My safety officer has told us that we can dispose of glass slides in the > >regular trash, because the tissue is fixed and considered no longer > >hazardous. > > > >Terri Clarke > > 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 ------- End of Original Message ------- From Eric <@t> ategra.com Wed Aug 3 10:43:28 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Histo Tech Mananger Supervisor and Bench Tech's needed for immediate openings Message-ID: Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisor, Histo Managers and Histo Bench Techs. These are fulltime permanent positions. Right now my Hottest jobs are in Colorado and Oregon who are seeking Lab Managers and Lab Supervisor. I have positions in California and Michigan who are seeking Histotech Supervisors. I also have Histo bench positions in Colorado,California,Pennsylvania,Illinois,New Hampshire,Michigan, Georgia,Florida,South Carolina, and Massachusetts. The client is currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recriuter 1-800-466-9919 ext 223 From Eric <@t> ategra.com Wed Aug 3 20:42:59 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Histo Tech Mananger Supervisor, Bench Tech's and Temps needed for immediate openings (this is not a duplicate) Message-ID: Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisor, Histo Managers and Histo Bench Techs. These are fulltime permanent positions. Right now my Hottest jobs are in Colorado and Oregon who are seeking Lab Managers and Lab Supervisor. I have positions in California and Michigan who are seeking Histotech Supervisors. I also have Histo bench positions in Colorado,California,Pennsylvania,Illinois,New Hampshire,Michigan, Georgia,Florida,South Carolina, and Massachusetts. I also have a temp position available in South Carolina at a Dermapathology lab. The assignment would be for 1 to 2 months and you must have dermpath experience.. This job has a deadline of 8/12/05. Please call ASAP if your interested.... There is also an opening for a Histotech traverler in Southern Florida. The client is currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recriuter 1-800-466-9919 ext 223 From meryl50 <@t> hotmail.com Thu Aug 4 02:31:57 2005 From: meryl50 <@t> hotmail.com (Meryl Roberts) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Problems with certification Message-ID: Hi there, I am a histologist in the UK with nearly 7 years experience, but I wish to move to the USA. However I'm having terrible problems finding work as it seems it's virtually impossible for me to become certified in the USA as none of my work experience is in an American lab- I've spoken to the ASCP and the NCA and they both say that I'm not elligeable to apply, and I'm currently waiting to hear back from the AMT (fingers crossed!).....is there any way I can become certified or should I just give up? I'm starting to get a bit despondant. sincerely, Meryl Roberts. From Mandy <@t> serotec.co.uk Thu Aug 4 02:50:35 2005 From: Mandy <@t> serotec.co.uk (Mandy Townsend) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Re: ooops, rat on rat Message-ID: Serotec are able to supply a polymer based kit that is suitable for use in rat on rat (and mouse on mouse) protocols - STAR4000A. Please contact our US office for further information. Serotec North & South America (website) 3200 Atlantic Avenue, Suite 105 Raleigh, NC 27604, USA Toll free: 1-800-265-7376 Fax: 919-878-3751 serotec@serotec-inc.com Product enquiry I hope this helps. Mandy Townsend MSc Technical Services Advisor Serotec Ltd 22 Bankside Station Approach Kidlington Oxfordshire OX5 1JE Tel: +44 1865 852733 Fax: +44 1865 852739 email: mandy@serotec.co.uk URL: http://www.serotec.com/ Serotec-Your first choice for antibodies! Join our free email update service IMPORTANT NOTICE: This message and any attachments may be confidential. If this has been sent to you in error, please contact the sender as soon as possible. Serotec Ltd. Registered in England No.1604642 Registered Office: Boswell House, 1-5 Broad Street, Oxford, OX1 SAW. UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle Callis Sent: Tuesday, August 02, 2005 9:21 PM To: Till, Renee; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: ooops, rat on rat You could use a biotinylated primary, as in the DAKO ARK methodology, eliminate the secondary to avoid having some "host" antiRat detect not only the rat primary, but also bind to rat tissue, then come back with a Strepavidin-HRP or AP. Sometimes you have to dilute out the primary more - I am not sure if anyone has a rat on rat kit, try Scytek out of Logan UT, you might get lucky on a kit. At 01:49 PM 8/2/2005, you wrote: >Okay, I knew I'd say that wrong. It is a rat anti-mouse antibody to be >used on rat tissues. How would you handle that? > > > >Renee' Till, HT > > > >Arkansas Childrens Nutrition Center > >1120 Marshall > >Little Rock, AR > >72202 > > > >(501) 364-2774 > > > >_______________________________________________ >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 e-mail has been scanned for all viruses by Star. The service is powered by MessageLabs. For more information on a proactive anti-virus service working around the clock, around the globe, visit: http://www.star.net.uk ________________________________________________________________________ From Kemlo.Rogerson <@t> elht.nhs.uk Thu Aug 4 02:13:14 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Re: Disposal of glass slides[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698E53@elht-exch1.xelht.nhs.uk> But what about confidentiality? You've got to grind them up first! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: 03 August 2005 17:31 To: Clarke, Mary; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Disposal of glass slides[Scanned] If you can dispose of slides in trash, how about large broken glass diposal box - safer for janitors and other personnel. At 09:20 AM 8/3/2005, you wrote: >My safety officer has told us that we can dispose of glass slides in the >regular trash, because the tissue is fixed and considered no longer >hazardous. > >Terri Clarke 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 Kemlo.Rogerson <@t> elht.nhs.uk Thu Aug 4 02:07:04 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] slide disposal[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698E50@elht-exch1.xelht.nhs.uk> Hire a cement mixer and get some lumps of metal or ball bearings. Put the slides and bearings in the mixer, turn on. Voila powdered glass; take to dump. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonnie Whitaker Sent: 02 August 2005 21:03 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] slide disposal[Scanned] Hi All, Can some of you give me ideas of how you handle old slide disposal? Particularly those of you in smaller, private labs. I have about 20 years worth of slides that I need to dispose of. If I have to send them with my biohazardous waste, I can, but it seems like an unnecessary expense. Currently that is what the procedure says that we do, but is it really necessary? Maybe someone out there would like to build a glass house : ) Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Thu Aug 4 02:09:42 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Whole Slide Imaging[Scanned] Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698E51@elht-exch1.xelht.nhs.uk> I'm sure you are correct but the initial bargaining went wrong when we sent an upper class twit with an attitude to the negotiating table; so I'm told. Never understood why we have less disposable earnings than the Americans AND everything costs more. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: 03 August 2005 17:25 To: Histonet Subject: Re: [Histonet] Whole Slide Imaging[Scanned] ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" "we in the UK tend to have to pay ?1 for $1 and I don't know why; History Channel told me it was about 'lend lease' and some unfair bargaining after the 2nd World War by the Americans; is that true?" When I was growing up in North London in the 50's and early 60's, the excuse for high prices was "we still have the debt from the first world war". I'm sure that in negotiations after WW2, the US bargained for its own national interest, as did the US (and Canada, and France, and .......). However, I don't think British economic policy has much to do with what happened during negotiations over who won the war 60 years ago, pre-EU. I suspect the real reason for British pound for dollar costs is commercial greed and taxes. Commercial greed is otherwise known as pricing according to "what the market will bear", or "gouge the buggers for every penny they've got". As for taxes, well... bureaucracy costs and the UK has one hell of a bureaucracy from what I recall before I emigrated. Bryan Llewellyn _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Thu Aug 4 07:49:22 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Whole Slide Imaging[Scanned] Message-ID: Rogerson Glad that you mentioned this. I have been in the USA for 3 years and during that time, the general attitude here that I have seen is that the USA gave the UK all the war goods. The fact that UK paid this all back several years ago never seemed to hit the headlines. As to why you have less, this is a complex question. Americans do, in general, have higher salaries, less expensive housing and living costs in many states. However, we do not have a national medical system. We have to pay one way or the other for medical care with our costs especially for medications that are far higher than Europe and most countries. Good new is that medical care if you can afford it is usually very good; bad news is that there are a lot of people here who cannot afford it. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rogerson Kemlo (ELHT) Pathology Sent: Thursday, August 04, 2005 2:10 AM To: Bryan Llewellyn; Histonet Subject: RE: [Histonet] Whole Slide Imaging[Scanned] I'm sure you are correct but the initial bargaining went wrong when we sent an upper class twit with an attitude to the negotiating table; so I'm told. Never understood why we have less disposable earnings than the Americans AND everything costs more. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: 03 August 2005 17:25 To: Histonet Subject: Re: [Histonet] Whole Slide Imaging[Scanned] ----- Original Message ----- From: "Rogerson Kemlo (ELHT) Pathology" "we in the UK tend to have to pay ?1 for $1 and I don't know why; History Channel told me it was about 'lend lease' and some unfair bargaining after the 2nd World War by the Americans; is that true?" When I was growing up in North London in the 50's and early 60's, the excuse for high prices was "we still have the debt from the first world war". I'm sure that in negotiations after WW2, the US bargained for its own national interest, as did the US (and Canada, and France, and .......). However, I don't think British economic policy has much to do with what happened during negotiations over who won the war 60 years ago, pre-EU. I suspect the real reason for British pound for dollar costs is commercial greed and taxes. Commercial greed is otherwise known as pricing according to "what the market will bear", or "gouge the buggers for every penny they've got". As for taxes, well... bureaucracy costs and the UK has one hell of a bureaucracy from what I recall before I emigrated. Bryan Llewellyn _______________________________________________ Histonet mailing list Histonet@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 specialstainsqueen <@t> hotmail.com Thu Aug 4 08:04:23 2005 From: specialstainsqueen <@t> hotmail.com (Sherri Anderson) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] cytospin preps and plus slides Message-ID: Dear Elizabeth- What is the overall volume of sample that you are actually receiving? If you are getting at least several mL of sample from your client, you might want to spin it down in a standard centrifuge prior to running it on the Cytospin -- this would allow you to concentrate the specimen and possibly use it in its entirety (i.e. spin it down, discard the supernatant, resuspend the pellet, and then pipette the sample into the sample chambers that are to be run on the Cytospin). Also, what type of sample chambers are you using on your Cytospin? For example, are you using the single white Cytofunnel, single brown Cytofunnel, double Cytofunnel, TPX chambers or Megafunnel? Also, what are your sample volumes that are being pipetted into the sample chambers? Each type of funnel has optimal volume ranges that should be observed. I would suggest referring back to the instructions for your particular type of funnel. You are using a standard cytology protocol (1000 RPM for 5 minutes) that should be sufficient. I would not recommend increasing the time (you don't want to "overspin" the cells and produce a drying artifact)...but you DO have a little room to "tweak" the RPM a bit. I would try increasing it slightly to 1100 or even 1200 and see what that does for you. If you feel that actual cell RECOVERY is not so much the issue and that it might be more of an issue with cellular ADHESION (to the slide during staining), would it be possible to get your client to use a fixative other than 10% NBF? When I worked in the lab, we often supplied doctors' offices with pre-filled collection cups in which they would subsequently send their samples to us. You would probably find greater success in terms of actual cellular adhesion if a cytology fixative containing Carbowax was utilized. This would allow you to actually let the slides air dry prior to staining (the Carbowax would coat the deposited cells and prevent any air-drying artifact). The combination of the Carbowax and allowing the slides to dry prior to running them through a staining protocol should significantly improve adhesion (as a wet slide will sometimes have some of its cells washed away during staining). These are just a few pointers and thoughts to ponder -- I hope that they help you. Sincerely, Sherri L. Anderson BS, HTL(ASCP) Product Specialist Anatomical Pathology Thermo Electron Corp. >From: "Elizabeth Chlipala" >To: "'Histonet'" >Subject: [Histonet] cytospin preps and plus slides >Date: Wed, 3 Aug 2005 12:42:30 -0600 > >Hello Everyone > >I'm working on a project for a client who is trying to isolate prostate >epithelial cells. I'm getting the cells submitted to me fixed in 10% >NBF in eppendorf tubes. I'm using a cytospin for 5 minutes at 1000 rpm >and plus slides. The problem is that I'm not getting the correct yield >of cells. They check the number of cells in the prep prior to me >receiving them. They are submitting numbers of cells in the thousands, >but when I prepare the cytospin slides, I'm only getting cells numbering >in the hundreds and sometimes less. Does anyone have any suggestions? > >Thanks in advance > >Liz > >Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC >Manager >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 _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From slappycraw <@t> yahoo.com Thu Aug 4 08:25:34 2005 From: slappycraw <@t> yahoo.com (Larry Woody) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Problems with certification In-Reply-To: Message-ID: <20050804132534.56873.qmail@web52610.mail.yahoo.com> Not sure who you talked to but there are plenty of places here in the US that will hire a tech without being certified. I can't tell you how many people I've worked with that were not certified, some good some not so good but keep trying and good luck. Meryl Roberts wrote:Hi there, I am a histologist in the UK with nearly 7 years experience, but I wish to move to the USA. However I'm having terrible problems finding work as it seems it's virtually impossible for me to become certified in the USA as none of my work experience is in an American lab- I've spoken to the ASCP and the NCA and they both say that I'm not elligeable to apply, and I'm currently waiting to hear back from the AMT (fingers crossed!).....is there any way I can become certified or should I just give up? I'm starting to get a bit despondant. sincerely, Meryl Roberts. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Barry.R.Rittman <@t> uth.tmc.edu Thu Aug 4 08:37:36 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Problems with certification Message-ID: In general I have found that Histotechs trained in the UK are highly regarded. The UK training is generally more formal with practical examinations that are extensive. Are you histo certified in the UK? If so I think that you will not have much of a problem finding a job here, USA certified or not. Have to be careful to research cost of living etc., as medical insurance takes a big bite. Also conditions of employment including career opportunities and type and quantity of work, supervisory role etc. Good luck Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Larry Woody Sent: Thursday, August 04, 2005 8:26 AM To: Meryl Roberts; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Problems with certification Not sure who you talked to but there are plenty of places here in the US that will hire a tech without being certified. I can't tell you how many people I've worked with that were not certified, some good some not so good but keep trying and good luck. Meryl Roberts wrote:Hi there, I am a histologist in the UK with nearly 7 years experience, but I wish to move to the USA. However I'm having terrible problems finding work as it seems it's virtually impossible for me to become certified in the USA as none of my work experience is in an American lab- I've spoken to the ASCP and the NCA and they both say that I'm not elligeable to apply, and I'm currently waiting to hear back from the AMT (fingers crossed!).....is there any way I can become certified or should I just give up? I'm starting to get a bit despondant. sincerely, Meryl Roberts. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Thu Aug 4 09:29:14 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] cytospin preps and plus slides Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698E5C@elht-exch1.xelht.nhs.uk> Maybe you need to spin faster and/or add protein to the vial to act as an adhesive; you may have better success with Cytyc LBC. Try the adhesive, bovine albumin, that may 'glue' ore of the cells onto the slide but I think you will always get cell loss with the filter paper. I found sedimentation techniques to be more successful. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: 03 August 2005 19:43 To: 'Histonet' Subject: [Histonet] cytospin preps and plus slides Hello Everyone I'm working on a project for a client who is trying to isolate prostate epithelial cells. I'm getting the cells submitted to me fixed in 10% NBF in eppendorf tubes. I'm using a cytospin for 5 minutes at 1000 rpm and plus slides. The problem is that I'm not getting the correct yield of cells. They check the number of cells in the prep prior to me receiving them. They are submitting numbers of cells in the thousands, but when I prepare the cytospin slides, I'm only getting cells numbering in the hundreds and sometimes less. Does anyone have any suggestions? Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 ajohnson <@t> aipathology.com Thu Aug 4 10:18:24 2005 From: ajohnson <@t> aipathology.com (Amy Johnson) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Negative controls Message-ID: <016A64931E1ED511B12C0002B3026080523368@SERV001> Our laboratory has been going thru the new Anatomic Pathology Checklist for CAP and we came across the Question "Are appropriate negative controls used?" We use to run one protocal for the negative mouse antibody and one protocol fro the negative rabbit antibody which was: no antigen retrieval no enzyme antibody incubation was only 2 minutes no amplification, no A/B block counterstain and post counterstain Now since we reviewed the CAP checklist we found out we were wrong, that we need to run the negative protocol the same as the primary antibody protocol. We have tried this and been having problems. We are getting positive staining in our negative controls even though there is no primary antibody being applied. How is this happening? How can this be solved? Thanks in advance for the help, Amy Johnson Associates in Pathology Wausau WI From steven.p.postl <@t> abbott.com Thu Aug 4 10:36:56 2005 From: steven.p.postl <@t> abbott.com (Steven P Postl) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Slide trays -wooden Message-ID: We are looking for slide trays that are made of wood. Remember those great trays that would hold 10 slides in each row, with 5 rows on each tray? Does any company out there still make them? Thanks. From TMcNemar <@t> lmhealth.org Thu Aug 4 10:35:40 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] basic auto stainer for paps Message-ID: <6CD94D97ED7D924BA5C2B588FA9528213967EA@mail.lmhealth.org> I'm looking to replace an older stainer. I'd like to get just a basic autostainer to use for PAPs. No bells or whistles but I'd like to run a basket of at least 60 slides at a time. I've just started looking but so far the ones I've found have far more features that I need, would use, or want to pay for. Anybody got any recommendations? Thanks, Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Hospital Newark, Ohio 43055 From tracy.bergeron <@t> crl.com Thu Aug 4 11:25:48 2005 From: tracy.bergeron <@t> crl.com (tracy.bergeron@crl.com) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Tissues for HTL Practical exam needed In-Reply-To: <016A64931E1ED511B12C0002B3026080523368@SERV001> Message-ID: HI All, I live in New Hampshire, and work in Wilmington, MA, and am looking to get a hold of human tissue for my HTL practical exam. I work in a research facility so I only have access to rodent tissue. Below is a list of the tissues I am looking to get a hold of as well as the final processed size each of them must be. Tissue Final processed size Uterus 1.5x1.5 square Cervix 1.5cm length Appendix Complete XC Ovary 1.0x1.0 square Any thoughts on the easiest way to access these tissues??? Responses can be private... Thank you in advance. Tracy E. Bergeron, BS, HT (ASCP) Histotechnician Charles River Laboratories Wilmington, MA 978-658-6000 x 1229 From Eric <@t> ategra.com Tue Aug 2 21:28:38 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Histo Tech Mananger Supervisor and Bench Tech's needed for immediate openings Message-ID: Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisor, Histo Managers and Histo Bench Techs. These are fulltime permanent positions. Right now my Hottest jobs are in Colorado and Oregon who are seeking Lab Managers and Lab Supervisor. I have positions in California and Michigan who are seeking Histotech Supervisors. I also have Histo bench positions in Colorado,California,Pennsylvania,Illinois,New Hampshire,Michigan, Georgia,Florida,South Carolina, and Massachusetts. The client is currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recriuter 1-800-466-9919 ext 223 From vd38 <@t> georgetown.edu Thu Aug 4 11:49:51 2005 From: vd38 <@t> georgetown.edu (Vernon Dailey) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] VEGF-R-2 Message-ID: <42F2472F.4050009@georgetown.edu> Hi Histonet, I am planning to do an IHC for the VEGF-receptor-2 antigen. I do not have much information about a good antibody for this project and was wondering if anyone could recommend a vendor/clone. I am planning on using the antibody to stain endothelial cells of a blood vessel in a mouse mammary tumor using vector's ABC kit to visualize. Thanks in advance for your help. -Vernon From pruegg <@t> ihctech.net Thu Aug 4 12:05:49 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Problems with certification In-Reply-To: Message-ID: <200508041705.j74H5mpf016595@chip.viawest.net> If you are certified in the UK as a histotech, I would think that you would not have any trouble getting a job in the US, as Barry points out the UK certification is usually much more in deapth than US. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R Sent: Thursday, August 04, 2005 6:38 AM To: histonet Subject: RE: [Histonet] Problems with certification In general I have found that Histotechs trained in the UK are highly regarded. The UK training is generally more formal with practical examinations that are extensive. Are you histo certified in the UK? If so I think that you will not have much of a problem finding a job here, USA certified or not. Have to be careful to research cost of living etc., as medical insurance takes a big bite. Also conditions of employment including career opportunities and type and quantity of work, supervisory role etc. Good luck Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Larry Woody Sent: Thursday, August 04, 2005 8:26 AM To: Meryl Roberts; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Problems with certification Not sure who you talked to but there are plenty of places here in the US that will hire a tech without being certified. I can't tell you how many people I've worked with that were not certified, some good some not so good but keep trying and good luck. Meryl Roberts wrote:Hi there, I am a histologist in the UK with nearly 7 years experience, but I wish to move to the USA. However I'm having terrible problems finding work as it seems it's virtually impossible for me to become certified in the USA as none of my work experience is in an American lab- I've spoken to the ASCP and the NCA and they both say that I'm not elligeable to apply, and I'm currently waiting to hear back from the AMT (fingers crossed!).....is there any way I can become certified or should I just give up? I'm starting to get a bit despondant. sincerely, Meryl Roberts. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@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 ajohnson <@t> aipathology.com Thu Aug 4 12:13:27 2005 From: ajohnson <@t> aipathology.com (Amy Johnson) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] More questions about negative controls Message-ID: <016A64931E1ED511B12C0002B302608052336B@SERV001> Do you think the timing of the negative control antibody has to be the same.....That is how we made the negative antibody protocol. Thanks Amy Johnson From esmith <@t> ikonisys.com Thu Aug 4 12:31:05 2005 From: esmith <@t> ikonisys.com (Emily Smith) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] cytospin preps and plus slides In-Reply-To: <200508041711.j74HB2cm029763@inbound-mx8.atl.registeredsite.com> Message-ID: Hello HistoNetters! What percentage cell recovery is expected/observed/acceptable for cytospin or for sedimentation techniques? Thank you! ~Emily Emily Smith Ikonisys Inc. 5 Science Park New Haven, CT 06511 ---------------------------------------------------- Message: 17 Date: Thu, 4 Aug 2005 15:29:14 +0100 From: "Rogerson Kemlo \(ELHT\) Pathology" Subject: RE: [Histonet] cytospin preps and plus slides To: "Elizabeth Chlipala" , "Histonet" Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698E5C@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" Maybe you need to spin faster and/or add protein to the vial to act as an adhesive; you may have better success with Cytyc LBC. Try the adhesive, bovine albumin, that may 'glue' ore of the cells onto the slide but I think you will always get cell loss with the filter paper. I found sedimentation techniques to be more successful. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: 03 August 2005 19:43 To: 'Histonet' Subject: [Histonet] cytospin preps and plus slides Hello Everyone I'm working on a project for a client who is trying to isolate prostate epithelial cells. I'm getting the cells submitted to me fixed in 10% NBF in eppendorf tubes. I'm using a cytospin for 5 minutes at 1000 rpm and plus slides. The problem is that I'm not getting the correct yield of cells. They check the number of cells in the prep prior to me receiving them. They are submitting numbers of cells in the thousands, but when I prepare the cytospin slides, I'm only getting cells numbering in the hundreds and sometimes less. Does anyone have any suggestions? Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 NoraBRL <@t> aol.com Thu Aug 4 12:55:01 2005 From: NoraBRL <@t> aol.com (NoraBRL@aol.com) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] RE: Wooden Slide Trays Message-ID: <213.62d0226.3023b075@aol.com> Brain Research Laboratories carries the wooden slide trays you are looking for, catalog #2550. Visit www.brainresearchlab.com for more information or call us at 1(888)BRL-5544. Nora Richards Brain Research Laboratories 1(888)BRL-5544 toll-free (US & Canada) 1(617)965-5544 tele steven.p.postl@abbott.com posted: We are looking for slide trays that are made of wood.? Remember those great trays that would hold 10 slides in each row, with 5 rows on each tray?? Does any company out there still make them?? Thanks. From ander093 <@t> tc.umn.edu Thu Aug 4 14:48:39 2005 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] looking for Ford Royer Message-ID: <6.2.3.4.0.20050804144736.01df1508@ander093.email.umn.edu> Does anyone have contact information for Ford Royer?? Need to get in touch with him. Thanks in advance. LuAnn From froyer <@t> bitstream.net Thu Aug 4 15:10:06 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] looking for Ford Royer In-Reply-To: <6.2.3.4.0.20050804144736.01df1508@ander093.email.umn.edu> Message-ID: I have recently changed companies and therefore address & phone... FORD M. ROYER Refurbished & New Histology Equipment Minnesota Medical Specialists, Inc. (aka: MN-Med) 7177 Madison Ave. Golden Valley, MN 55427 Phone: 888-790-9686 (toll free) 763-542-8725 (local) Fax: 763-546-4830 Email: URL: Sorry for the inconvenience, LuAnn (and others)... ~ Ford -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of LuAnn Anderson Sent: Thursday, August 04, 2005 2:49 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] looking for Ford Royer Does anyone have contact information for Ford Royer?? Need to get in touch with him. Thanks in advance. LuAnn _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dharclerode <@t> cytoritx.com Thu Aug 4 15:53:33 2005 From: dharclerode <@t> cytoritx.com (Donna Harclerode) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] FW: RE Slide trays wooden Message-ID: -----Original Message----- From: Donna Harclerode Sent: Thursday, August 04, 2005 1:43 PM To: 'histonet@lists.utsouthwestern.edu'' Subject: RE Slide trays wooden Hi Steven You can get the wooden trays from Brain Research Laboratories. They call them "slide folders". They have all kind of useful items- Oversized slides and coverglasses, nets for staining free floating sections for IHC. They also sell good quality paint brushes for low prices. They are a small and very good company for specialty items. Brain Research Laboratories Division of Cambridge Intelligent Systems, Inc. Waban P.O. Box 88 Newton, MA 02468 Please use this for mailing orders as well as remittance Telephone: (617)965-5544 Toll-Free: (888)BRL-5544 (USA & Canada) Fax: (617)965-6220 E-mail: brl@brainresearchlab.com Donna Harclerode, HT, (ASCP), HTL, QIHC Immunohistochemist Cytori Therapeutics 6740 Top Gun St. San Diego, CA 92121 858-458-0900 ext 322 dharclerode@cytoritx.com Date: Thu, 4 Aug 2005 10:36:56 -0500 From: "Steven P Postl" Subject: [Histonet] Slide trays -wooden To: histonet@lists.utsouthwestern.edu We are looking for slide trays that are made of wood. Remember those great trays that would hold 10 slides in each row, with 5 rows on each tray? Does any company out there still make them? Thanks. From mtitford <@t> aol.com Thu Aug 4 16:15:44 2005 From: mtitford <@t> aol.com (mtitford@aol.com) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" histotechs working in USA Message-ID: <8C76750CF43CABD-1340-1735@MBLK-M32.sysops.aol.com> Meryl Roberts somewhere in the U.K. asks about getting a job in the USA. I don't think a histotechnologist from the United Kingdom would have any trouble finding a job in the USA, but there would be other problems. Two come to mind. 1) If you wish to emigrate to the USA from the U.K., the government here will not want you to enter the country just because you have a job offer. They want to be sure there are no unemployed histotechnologists here who want the job first. An exception would be if you have a skill no one else in the USA has.Other exceptions are if you are married to an American, etc 2) The second problem is fully qualified histotechnologists in the U.K. have been through a degree level program, are state registered there and are called "Clinical Laboratory Scientists" or something similier. They are trained in all sections of clinical laboratory medicine. They might consider themselves over qualified (and maybe underpaid!) for many histo jobs in the USA. (and then be unhappy!). Mike Titford Pathology USA Pathology Mobile AL USA From Adesupod <@t> aol.com Thu Aug 4 17:06:39 2005 From: Adesupod <@t> aol.com (Adesupod@aol.com) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" histotechs working in USA Message-ID: <19a.3931fe60.3023eb6f@aol.com> Hi Mike, I quite very much agree with your response. Brit histotechs are highly trained in all tha Medical Laboratory disciplines, and they are being referred to as Clinical Lab. Scientists. Taking up jobs as histotechs in the United States by the Brit histotechs will not earn them much pay. I know this because I am a trained Clinical Lab.Scientist with specialization in Histology and Cytology( 4-years degree program + internship). I am currently working in the United States as an Histotechnologist. Banjo Adesuyi. From conniemoss <@t> relia.net Fri Aug 5 00:50:12 2005 From: conniemoss <@t> relia.net (Connie McManus) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Negative controls In-Reply-To: <016A64931E1ED511B12C0002B3026080523368@SERV001> References: <016A64931E1ED511B12C0002B3026080523368@SERV001> Message-ID: <49461.208.186.240.165.1123221012.squirrel@email.relia.net> Amy, I missed what antibody you're talking about and what detection system you're using, but in general, this is how I was taught to treat assays that use a neg ab ctrl: The negative antibody must be run with the same protocol as the pos antibody and is diluted to the same concentration. Treat positive and negative ctrl tissues with pos and neg antibodies (use the same tissue ctrls for both abs, each ctrl slide has pos and neg tissue. I embed the pos and neg tissues in the same block and orient the neg ctrl to the top of the slide so I would always know which tissue was which). If you are getting staining with your negative antibody, it could be background staining. This may be alleviated by... 1- check the blocking: a/b block, h202 or general protein blocking (if you're using Alk Phos, you may need to treat the sections with levamisole - h202 is not needed). 2- antigent retrieval timing needs to be revised (if you are using a/r) -- you may need to find a balance between the timing that makes a good pos rxn in the pos ab and no background in the neg ab. 3 - Be sure the detection system is compatible with your tissues -- i.e. don't use alk phos on tissues rich in endogenous alk phos and the same with peroxidase. The other possibility is that the neg ab ctrl may be contaminated. To avoid this, I always aliquotted small volumes into cryo vials and stored them as per manufacturer's instructions. Each new lot of antibody is qc'd along with titering to assure the working dilution is acceptable. Neg ab ctrls are run alongside the pos ab at this time. Records of the results are filed. I hope I covered all the bases here. I may have forgotten a thing or two, but I'm sure if I did leave something out, there are those who will bring it up. I think, in a nutshell, the basic rule of thumb is to treat the neg ab ctrl exactly the same as the pos ab. hope this has been helpful. -- Connie McManus Mt Ogden Scientific Services -Providing full service electron microscopy for you 950 W Kershaw, Suite E Ogden UT 84401 toll-free: 877/311-6677 direct: 801/745-2583 cell: 435/757/2975 fax: 435/514-1781 conniemoss@relia.net www.mtogdensci.com ====== Amy Johnson said: > Our laboratory has been going thru the new Anatomic Pathology Checklist for > CAP and we came across > the Question "Are appropriate negative controls used?" We use to run one > protocal for the negative > mouse antibody and one protocol fro the negative rabbit antibody which was: > > no antigen retrieval > no enzyme > antibody incubation was only 2 minutes > no amplification, no A/B block > counterstain and post counterstain > Now since we reviewed the CAP checklist we found out we were wrong, that we > need to run the negative > protocol the same as the primary antibody protocol. We have tried this and > been having problems. We are getting > positive staining in our negative controls even though there is no primary > antibody being applied. How is this happening? > How can this be solved? > Thanks in advance for the help, > Amy Johnson > Associates in Pathology > Wausau WI > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Kemlo.Rogerson <@t> elht.nhs.uk Fri Aug 5 02:16:20 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] cytospin preps and plus slides Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698E62@elht-exch1.xelht.nhs.uk> Haven't a clue but I bet you loose more than half. If it's urine even more; I know platelets and rbcs haven't got a prayer in staying on in a fluid devoid of protein. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Emily Smith Sent: 04 August 2005 18:31 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cytospin preps and plus slides Hello HistoNetters! What percentage cell recovery is expected/observed/acceptable for cytospin or for sedimentation techniques? Thank you! ~Emily Emily Smith Ikonisys Inc. 5 Science Park New Haven, CT 06511 ---------------------------------------------------- Message: 17 Date: Thu, 4 Aug 2005 15:29:14 +0100 From: "Rogerson Kemlo \(ELHT\) Pathology" Subject: RE: [Histonet] cytospin preps and plus slides To: "Elizabeth Chlipala" , "Histonet" Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698E5C@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" Maybe you need to spin faster and/or add protein to the vial to act as an adhesive; you may have better success with Cytyc LBC. Try the adhesive, bovine albumin, that may 'glue' ore of the cells onto the slide but I think you will always get cell loss with the filter paper. I found sedimentation techniques to be more successful. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: 03 August 2005 19:43 To: 'Histonet' Subject: [Histonet] cytospin preps and plus slides Hello Everyone I'm working on a project for a client who is trying to isolate prostate epithelial cells. I'm getting the cells submitted to me fixed in 10% NBF in eppendorf tubes. I'm using a cytospin for 5 minutes at 1000 rpm and plus slides. The problem is that I'm not getting the correct yield of cells. They check the number of cells in the prep prior to me receiving them. They are submitting numbers of cells in the thousands, but when I prepare the cytospin slides, I'm only getting cells numbering in the hundreds and sometimes less. Does anyone have any suggestions? Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 Kemlo.Rogerson <@t> elht.nhs.uk Fri Aug 5 02:53:38 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" histotechs working in USA Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4CE@elht-exch1.xelht.nhs.uk> To be precise most 'qualified histotechnologists' have a post graduate qualification; mostly MSc but a few PHD's. We are called Biomedical Scientists and some Advanced Practitioners. My question is, are histo jobs in the USA different from those in the UK. If they are the same then are UK BMS's overqualified for these jobs in the UK too? If they different then how? How can 'less qualified' Staff carry out these tasks seemingly very well in the US but it needs a postgraduate qualification in the UK. Clinical Scientists are another grade of degree entry Scientist who usually enter with a science degree, get an MSc then a PHD and are usually paid more than the BMS. In many cases they can gain entry to the appropriate Royal College and head up a Department. The difference? Dunno! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of mtitford@aol.com Sent: 04 August 2005 22:16 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] "Brit" histotechs working in USA Meryl Roberts somewhere in the U.K. asks about getting a job in the USA. I don't think a histotechnologist from the United Kingdom would have any trouble finding a job in the USA, but there would be other problems. Two come to mind. 1) If you wish to emigrate to the USA from the U.K., the government here will not want you to enter the country just because you have a job offer. They want to be sure there are no unemployed histotechnologists here who want the job first. An exception would be if you have a skill no one else in the USA has.Other exceptions are if you are married to an American, etc 2) The second problem is fully qualified histotechnologists in the U.K. have been through a degree level program, are state registered there and are called "Clinical Laboratory Scientists" or something similier. They are trained in all sections of clinical laboratory medicine. They might consider themselves over qualified (and maybe underpaid!) for many histo jobs in the USA. (and then be unhappy!). Mike Titford Pathology USA Pathology Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MLafrini <@t> csmlab.com Fri Aug 5 07:34:47 2005 From: MLafrini <@t> csmlab.com (Michael LaFriniere) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Seeking Co Pah information Message-ID: I manage a private laboratory and currently searching for a new billing system (3rd party). I am seeking a package that works well with Co path/Mysis pathology computer system. If anyone has any suggestions or experience with billing packages with Co path/Mysis, I would really appreciate the information... Thank you Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax MLafrini@csmlab.com From Diane.Gladney <@t> se.amedd.army.mil Fri Aug 5 09:59:37 2005 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Tzanck Stain Proficiency Testing Message-ID: <4D55B2E997EFAE4DA6081DDE100B83026DD278@amedmlsermc133.amed.ds.army.mil> Hi Fellow Histonetters, We have a new Dermatologist that wants to perform Tzanck Stain on skin smears in his clinic. Out QA/QC manager asked me to find out if there is a Proficiency Test available (like a survey from CAP) for this stain. We are looking to get the dermatologist his own CLIP certificate and this stain is considered moderate complexity testing. I am not even sure if a proficiency test even exists for this stain. Does anyone out there in Histonet Land have any info on this or some guidance that I may share with our QA/QC manager? As always, I know that if it is available, someone on Histonet will know. Thanks in advance for your help. Diane Diane C. Gladney, HT (ASCP) Supervisor, Histology/Cytology Moncrief Army Community Hospital Dept. of Pathology 4500 Stuart Street P.O. Box 484 Ft. Jackson, SC 29207 Email: diane.gladney@se.amedd.army.mil Phone: 803-751-2530 FAX: 803-751-7829 From tp2 <@t> medicine.wisc.edu Fri Aug 5 10:47:22 2005 From: tp2 <@t> medicine.wisc.edu (Thomas Pier) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] troubles with CD4 Message-ID: Hello all, I was wondering if anyone could help me with some problems that I'm having with staining for CD4. My control tissue for the work up is formalin fixted, paraffin embeded tonsil. I'm using Labvision's CD4 Ab-2 clone 1F6. I have diluted it anywhere from 1:10-1:50. I'm using 1% goat serum in TBS as my diluent. I've tried EDTA HIER as recomended on the data sheet. I've also tried protease XXV w/ or w/o the EDTA. I shouldn't be running into the peroxidase problem since I'm using an alkaline phosphatase detection system (Biocare's Vulcan Fast Red). I have also tried using or not using a 10% goat serum protein block. I have gotten some staining, but nowhere near as much as I should in a tonsil. Any help that you could give me would be greatly appreciated. Tom Pier From emerald_lake77 <@t> yahoo.com Fri Aug 5 11:39:33 2005 From: emerald_lake77 <@t> yahoo.com (GT Hebert) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] F4/80 Ab clone CI:A3-1 not working for foam cells - why? Message-ID: <20050805163933.81444.qmail@web31711.mail.mud.yahoo.com> Hello again, I just worked out my F4/80 antibody on mouse tissue. The mouse tissue, aortic arch with athrosclerotic lesions came up negative though. I would think that the foam cells (macrophages??) would light up as the do when I stain with CD68 ... but only 1 or 2 cells outside the lesion lit up. The Kupffer cells in the liver and the red pulp in the spleen (my + controls) came out great and the negative isotype controls on all samples are clear. Any idea why this may be happening? I have seen this antibody used on a number of literature papers staining foam cells in AT lesions (same clone - just different dilutions (1:25, 1:200m etc -- all paraffin) -- and they showed staining of foam cells. My tissues are all treated the same: 24 hour fixation in 4%PF, routine processed and embedded, 4 micron sections, deparaffinized ... and then IHC (optimal conditions). I'm sort of lost with this one ... I know the antibody works. Just the results are not what I expected. Any information would be greatly appreciated. Gustave Hebert Wyeth Cambridge --------------------------------- Start your day with Yahoo! - make it your home page From mtitford <@t> aol.com Fri Aug 5 11:58:47 2005 From: mtitford <@t> aol.com (mtitford@aol.com) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: <8C767F6149E95D5-1544-22F7@MBLK-M23.sysops.aol.com> Rogerson Kemlo carries on the discussion about histotechnologist training. In regards to hospital histology labs in the USA: I don't want to start a firestorm of criticism here, but I think there is a real danger that in Great Britain those technologists performing in the histology laboratories may be over qualified.A few months ago here on the Histonet, someone in the U.K. was bemoaning the lack of histology technologists in the U.K., and how short staffed they were. Was that because the bar had been set too high? Was the level of education required too much for the compensation received? Has a "closed shop" (to use a union term!) in effect, been created? Here in the United States there are hundreds of community type hospitals and smaller medical centers that have histology laboratories. In those laboratories work histotechnologists doing a fine job with a lot less education than a fully qualified histotechnologist in the U.K.. At the other end of the spectrum are histology laboratories here doing more advanced procedures where degrees and extra training are required. However, histotechnology training in the U.K. and USA are inherently different. When I lived in the U.K. years and years ago (and it may have changed now), all the hospitals were government owned, lab personnel got a day off each week with pay to attend IMLT classes, and the classes were free. To work in a hospital laboratory you had to be "State Registered". In the USA hospitals are owned by all sorts of organizations. You don't get "day release" to attend classes. If you do attend classes, money has to come from somewhere to pay for it. Further you may live miles/hours from the nearest college where you can attend classes.Since the different organisations may be "for profit" or even "Non-profit", they want to save money by not paying too higher salaries. Most often they pay what other hospitals in the area pay for the same type work. The 50 states have different requirements for working in their hospital laboratories. Hospitals have coped with these differences by making histology laboratories a separate section of the laboratory, with technicians/technologists trained just to work in that lab. I think a histotechnologist from the U.K. could easily find a position in the USA, but they might feel underappreciated and underpaid! (maybe even under challanged!) Well, that's my two bits worth! Mike Titford USA Pathology Mobile AL USA From slappycraw <@t> yahoo.com Fri Aug 5 12:35:08 2005 From: slappycraw <@t> yahoo.com (Larry Woody) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? In-Reply-To: <8C767F6149E95D5-1544-22F7@MBLK-M23.sysops.aol.com> Message-ID: <20050805173508.81307.qmail@web52606.mail.yahoo.com> You may want to try some of the big biotech companies where there is a need for people with more skills in the lab and the compensation can be better than a medical lab. mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion about histotechnologist training. In regards to hospital histology labs in the USA: I don't want to start a firestorm of criticism here, but I think there is a real danger that in Great Britain those technologists performing in the histology laboratories may be over qualified.A few months ago here on the Histonet, someone in the U.K. was bemoaning the lack of histology technologists in the U.K., and how short staffed they were. Was that because the bar had been set too high? Was the level of education required too much for the compensation received? Has a "closed shop" (to use a union term!) in effect, been created? Here in the United States there are hundreds of community type hospitals and smaller medical centers that have histology laboratories. In those laboratories work histotechnologists doing a fine job with a lot less education than a fully qualified histotechnologist in the U.K.. At the other end of the spectrum are histology laboratories here doing more advanced procedures where degrees and extra training are required. However, histotechnology training in the U.K. and USA are inherently different. When I lived in the U.K. years and years ago (and it may have changed now), all the hospitals were government owned, lab personnel got a day off each week with pay to attend IMLT classes, and the classes were free. To work in a hospital laboratory you had to be "State Registered". In the USA hospitals are owned by all sorts of organizations. You don't get "day release" to attend classes. If you do attend classes, money has to come from somewhere to pay for it. Further you may live miles/hours from the nearest college where you can attend classes.Since the different organisations may be "for profit" or even "Non-profit", they want to save money by not paying too higher salaries. Most often they pay what other hospitals in the area pay for the same type work. The 50 states have different requirements for working in their hospital laboratories. Hospitals have coped with these differences by making histology laboratories a separate section of the laboratory, with technicians/technologists trained just to work in that lab. I think a histotechnologist from the U.K. could easily find a position in the USA, but they might feel underappreciated and underpaid! (maybe even under challanged!) Well, that's my two bits worth! Mike Titford USA Pathology Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Start your day with Yahoo! - make it your home page From dmccaig <@t> ckha.on.ca Fri Aug 5 12:41:16 2005 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: <3E5A3F039F0BD8118B4700C00D002024043634@CKHA9> What are you all using to compare the wage scales. Biomedical scientists do not get paid as well as we do in North America if you factor in their cost of living. I understand there is a recent trend to increase the wages in order to keep them on board as there is a lot of migration to various labs throughout a career where in NA we tend to stay put. Diana McCaig, MLT -----Original Message----- From: Larry Woody [SMTP:slappycraw@yahoo.com] Sent: Friday, August 05, 2005 1:35 PM To: mtitford@aol.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? You may want to try some of the big biotech companies where there is a need for people with more skills in the lab and the compensation can be better than a medical lab. mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion about histotechnologist training. In regards to hospital histology labs in the USA: I don't want to start a firestorm of criticism here, but I think there is a real danger that in Great Britain those technologists performing in the histology laboratories may be over qualified.A few months ago here on the Histonet, someone in the U.K. was bemoaning the lack of histology technologists in the U.K., and how short staffed they were. Was that because the bar had been set too high? Was the level of education required too much for the compensation received? Has a "closed shop" (to use a union term!) in effect, been created? Here in the United States there are hundreds of community type hospitals and smaller medical centers that have histology laboratories. In those laboratories work histotechnologists doing a fine job with a lot less education than a fully qualified histotechnologist in the U.K.. At the other end of the spectrum are histology laboratories here doing more advanced procedures where degrees and extra training are required. However, histotechnology training in the U.K. and USA are inherently different. When I lived in the U.K. years and years ago (and it may have changed now), all the hospitals were government owned, lab personnel got a day off each week with pay to attend IMLT classes, and the classes were free. To work in a hospital laboratory you had to be "State Registered". In the USA hospitals are owned by all sorts of organizations. You don't get "day release" to attend classes. If you do attend classes, money has to come from somewhere to pay for it. Further you may live miles/hours from the nearest college where you can attend classes.Since the different organisations may be "for profit" or even "Non-profit", they want to save money by not paying too higher salaries. Most often they pay what other hospitals in the area pay for the same type work. The 50 states have different requirements for working in their hospital laboratories. Hospitals have coped with these differences by making histology laboratories a separate section of the laboratory, with technicians/technologists trained just to work in that lab. I think a histotechnologist from the U.K. could easily find a position in the USA, but they might feel underappreciated and underpaid! (maybe even under challanged!) Well, that's my two bits worth! Mike Titford USA Pathology Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Start your day with Yahoo! - make it your home page _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histo112000 <@t> yahoo.com Fri Aug 5 13:25:32 2005 From: histo112000 <@t> yahoo.com (Ryan Yvonne) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] negative controls Message-ID: <20050805182533.38308.qmail@web50609.mail.yahoo.com> Amy, are you running commercial or novel antibodies? You're secondary may not be clean also. I've had problems with a dirty secondary causing positive staining, I got a different secondary and it was fine. To have a true negative control, you do need to run the same protocol as the positive or the tissue isn't going through the same solutions and can't be considered a true negative. Yvonne histonet-request@lists.utsouthwestern.edu wrote: Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. cytospin preps and plus slides (Elizabeth Chlipala) 2. Casp-3 advice (9msm8@qlink.queensu.ca) 3. RE: fixation of fresh frozen section (Lewis, Sarah) 4. PLP fixation info RE: fixation of fresh frozen section (Gayle Callis) 5. Re: Re: Disposal of glass slides (cindipqr@wowway.com) 6. Histo Tech Mananger Supervisor and Bench Tech's needed for immediate openings (Eric Dye (ext 223)) 7. Histo Tech Mananger Supervisor, Bench Tech's and Temps needed for immediate openings (this is not a duplicate) (Eric Dye (ext 223)) 8. Problems with certification (Meryl Roberts) 9. RE: Re: ooops, rat on rat (Mandy Townsend) 10. RE: Re: Disposal of glass slides[Scanned] (Rogerson Kemlo (ELHT) Pathology) 11. RE: slide disposal[Scanned] (Rogerson Kemlo (ELHT) Pathology) 12. RE: Whole Slide Imaging[Scanned] (Rogerson Kemlo (ELHT) Pathology) 13. RE: Whole Slide Imaging[Scanned] (Rittman, Barry R) 14. RE: cytospin preps and plus slides (Sherri Anderson) 15. Re: Problems with certification (Larry Woody) 16. RE: Problems with certification (Rittman, Barry R) 17. RE: cytospin preps and plus slides (Rogerson Kemlo (ELHT) Pathology) 18. Negative controls (Amy Johnson) 19. Slide trays -wooden (Steven P Postl) 20. basic auto stainer for paps (Tom McNemar) 21. Tissues for HTL Practical exam needed (tracy.bergeron@crl.com) 22. Histo Tech Mananger Supervisor and Bench Tech's needed for immediate openings (Eric Dye (ext 223)) 23. VEGF-R-2 (Vernon Dailey) ---------------------------------------------------------------------- Message: 1 Date: Wed, 3 Aug 2005 12:42:30 -0600 From: "Elizabeth Chlipala" Subject: [Histonet] cytospin preps and plus slides To: "'Histonet'" Message-ID: <000001c5985b$1c3023d0$a7d48a80@AMY> Content-Type: text/plain; charset="us-ascii" Hello Everyone I'm working on a project for a client who is trying to isolate prostate epithelial cells. I'm getting the cells submitted to me fixed in 10% NBF in eppendorf tubes. I'm using a cytospin for 5 minutes at 1000 rpm and plus slides. The problem is that I'm not getting the correct yield of cells. They check the number of cells in the prep prior to me receiving them. They are submitting numbers of cells in the thousands, but when I prepare the cytospin slides, I'm only getting cells numbering in the hundreds and sometimes less. Does anyone have any suggestions? Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 ------------------------------ Message: 2 Date: Wed, 3 Aug 2005 14:53:35 -0400 (EDT) From: 9msm8@qlink.queensu.ca Subject: [Histonet] Casp-3 advice To: histonet@lists.utsouthwestern.edu Message-ID: <51618.192.75.165.28.1123095215.squirrel@qlink.queensu.ca> Content-Type: text/plain;charset=iso-8859-1 Hello Histonet. I am a student in a lab where histologists are rare to non-existant. I am working with 5 micrometer thick mouse hippocampal parrafin embedded sections (I used paraformaldehyde) and I want to do an immunohistochemical stain for casp-3. I am using the marker as a detection for relatively early apoptosis. The QUESTION I am asking is: Should I be using this method with a fluorescence signal or a horshradish peroxidase counterstain? Given that I probably will want my analysis to be more of a qualitative descritpion. What should I be aware of before making my decision? What are the advantages and disadvantages? Any help will be greatly appreciated. ------------------------------ Message: 3 Date: Wed, 3 Aug 2005 15:32:00 -0400 From: "Lewis, Sarah" Subject: RE: [Histonet] fixation of fresh frozen section To: "Edmondson David \(RBV\) NHS Christie Tr" , "HISTONET \(E-mail\)" Message-ID: <714B9F12B4E18C4C843B66E8E190F2AD447562@res2k3ms01.CRII.ORG> Content-Type: text/plain; charset="iso-8859-1" I would love more information on this subject. Thanks in advance!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net -----Original Message----- From: Edmondson David (RBV) NHS Christie Tr [mailto:David.Edmondson@christie-tr.nwest.nhs.uk] Sent: Wednesday, August 03, 2005 12:37 PM To: Bruijntjes, J.P. Cc: Histonet (E-mail 2) Subject: RE: [Histonet] fixation of fresh frozen section Hi I recently heard a presentation where they spoke of Periodate-Lysine-Paraformaldehyde in the context of Frozen sections. The coagulant fixatives do not alter the proteins chemistry so on might suggest no loss of antigenicity, the "Gold Standard". But the morphology of PLP is much better. It is fifteen and more years since I used it myself and maybe a search would be a quicker way to find the concentrations. I ahve an email contact if you need to ask him. David Christie Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bruijntjes, J.P. Sent: 03 August 2005 14:39 To: histonet@pathology.swmed.edu Subject: [Histonet] fixation of fresh frozen section Hi all When I do immunocytochemistry on fresh frozen section I fix most, if not all slides with acetone. Sometimes I use cold methanol because the datasheet tells me to do so. Is anyone of you who prefer other fixatives? Joost Bruijntjes TNO Quality of Life Zeist Holland This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Wed, 03 Aug 2005 14:55:57 -0600 From: Gayle Callis Subject: [Histonet] PLP fixation info RE: fixation of fresh frozen section To: "Lewis, Sarah" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050803134024.01b5c108@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Sarah, This was a previous message on Histonet concerning PLP fixation. We have great success with this for mouse and hamster perfusion fixation for a prion study then paraffin processing of tissues. The recipe for PLP has been put on Histonet previously, going to Histonet archives should access fixative recipe. I haven't had success with frozen section fixation, although some have done so, my impatience in taking time to work with it to do immunofluorescence work. Bob Chiovette wrote: 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 Good luck Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 5 Date: Wed, 3 Aug 2005 18:17:07 -0600 From: cindipqr@wowway.com Subject: Re: [Histonet] Re: Disposal of glass slides To: Gayle Callis , "Clarke, Mary" , Histonet@lists.utsouthwestern.edu Cc: histonet-bounces@lists.utsouthwestern.edu Message-ID: <20050803231204.M24570@wowway.com> Content-Type: text/plain; charset=iso-8859-1 Our facility likes them in smaller boxes, taped shut and labeled as glass slides for disposal. This way it's safe and not as heavy and bulky as the regular glass disposal boxes. Cindi HFH Detroit Mi ---------- Original Message ----------- From: Gayle Callis To: "Clarke, Mary" , Histonet@lists.utsouthwestern.edu Sent: Wed, 03 Aug 2005 10:30:38 -0600 Subject: [Histonet] Re: Disposal of glass slides > If you can dispose of slides in trash, how about large broken glass > diposal box - safer for janitors and other personnel. > > At 09:20 AM 8/3/2005, you wrote: > >My safety officer has told us that we can dispose of glass slides in the > >regular trash, because the tissue is fixed and considered no longer > >hazardous. > > > >Terri Clarke > > 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 ------- End of Original Message ------- ------------------------------ Message: 6 Date: Wed, 3 Aug 2005 11:43:28 -0400 From: Eric Dye (ext 223) Subject: [Histonet] Histo Tech Mananger Supervisor and Bench Tech's needed for immediate openings To: Histonetters Message-ID: Content-Type: text/plain Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisor, Histo Managers and Histo Bench Techs. These are fulltime permanent positions. Right now my Hottest jobs are in Colorado and Oregon who are seeking Lab Managers and Lab Supervisor. I have positions in California and Michigan who are seeking Histotech Supervisors. I also have Histo bench positions in Colorado,California,Pennsylvania,Illinois,New Hampshire,Michigan, Georgia,Florida,South Carolina, and Massachusetts. The client is currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recriuter 1-800-466-9919 ext 223 ------------------------------ Message: 7 Date: Wed, 3 Aug 2005 21:42:59 -0400 From: Eric Dye (ext 223) Subject: [Histonet] Histo Tech Mananger Supervisor, Bench Tech's and Temps needed for immediate openings (this is not a duplicate) To: Histonetters Message-ID: Content-Type: text/plain Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisor, Histo Managers and Histo Bench Techs. These are fulltime permanent positions. Right now my Hottest jobs are in Colorado and Oregon who are seeking Lab Managers and Lab Supervisor. I have positions in California and Michigan who are seeking Histotech Supervisors. I also have Histo bench positions in Colorado,California,Pennsylvania,Illinois,New Hampshire,Michigan, Georgia,Florida,South Carolina, and Massachusetts. I also have a temp position available in South Carolina at a Dermapathology lab. The assignment would be for 1 to 2 months and you must have dermpath experience.. This job has a deadline of 8/12/05. Please call ASAP if your interested.... There is also an opening for a Histotech traverler in Southern Florida. The client is currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recriuter 1-800-466-9919 ext 223 ------------------------------ Message: 8 Date: Thu, 04 Aug 2005 08:31:57 +0100 From: "Meryl Roberts" Subject: [Histonet] Problems with certification To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hi there, I am a histologist in the UK with nearly 7 years experience, but I wish to move to the USA. However I'm having terrible problems finding work as it seems it's virtually impossible for me to become certified in the USA as none of my work experience is in an American lab- I've spoken to the ASCP and the NCA and they both say that I'm not elligeable to apply, and I'm currently waiting to hear back from the AMT (fingers crossed!).....is there any way I can become certified or should I just give up? I'm starting to get a bit despondant. sincerely, Meryl Roberts. ------------------------------ Message: 9 Date: Thu, 4 Aug 2005 08:50:35 +0100 From: "Mandy Townsend" Subject: RE: [Histonet] Re: ooops, rat on rat To: "Gayle Callis" , "Till, Renee" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Serotec are able to supply a polymer based kit that is suitable for use in rat on rat (and mouse on mouse) protocols - STAR4000A. Please contact our US office for further information. Serotec North & South America (website) 3200 Atlantic Avenue, Suite 105 Raleigh, NC 27604, USA Toll free: 1-800-265-7376 Fax: 919-878-3751 serotec@serotec-inc.com Product enquiry I hope this helps. Mandy Townsend MSc Technical Services Advisor Serotec Ltd 22 Bankside Station Approach Kidlington Oxfordshire OX5 1JE Tel: +44 1865 852733 Fax: +44 1865 852739 email: mandy@serotec.co.uk URL: http://www.serotec.com/ Serotec-Your first choice for antibodies! Join our free email update service IMPORTANT NOTICE: This message and any attachments may be confidential. If this has been sent to you in error, please contact the sender as soon as possible. Serotec Ltd. Registered in England No.1604642 Registered Office: Boswell House, 1-5 Broad Street, Oxford, OX1 SAW. UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle Callis Sent: Tuesday, August 02, 2005 9:21 PM To: Till, Renee; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: ooops, rat on rat You could use a biotinylated primary, as in the DAKO ARK methodology, eliminate the secondary to avoid having some "host" antiRat detect not only the rat primary, but also bind to rat tissue, then come back with a Strepavidin-HRP or AP. Sometimes you have to dilute out the primary more - I am not sure if anyone has a rat on rat kit, try Scytek out of Logan UT, you might get lucky on a kit. At 01:49 PM 8/2/2005, you wrote: >Okay, I knew I'd say that wrong. It is a rat anti-mouse antibody to be >used on rat tissues. How would you handle that? > > > >Renee' Till, HT > > > >Arkansas Childrens Nutrition Center > >1120 Marshall > >Little Rock, AR > >72202 > > > >(501) 364-2774 > > > >_______________________________________________ >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 e-mail has been scanned for all viruses by Star. The service is powered by MessageLabs. For more information on a proactive anti-virus service working around the clock, around the globe, visit: http://www.star.net.uk ________________________________________________________________________ ------------------------------ Message: 10 Date: Thu, 4 Aug 2005 08:13:14 +0100 From: "Rogerson Kemlo \(ELHT\) Pathology" Subject: RE: [Histonet] Re: Disposal of glass slides[Scanned] To: "Gayle Callis" , "Clarke, Mary" , Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698E53@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" But what about confidentiality? You've got to grind them up first! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: 03 August 2005 17:31 To: Clarke, Mary; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Disposal of glass slides[Scanned] If you can dispose of slides in trash, how about large broken glass diposal box - safer for janitors and other personnel. At 09:20 AM 8/3/2005, you wrote: >My safety officer has told us that we can dispose of glass slides in the >regular trash, because the tissue is fixed and considered no longer >hazardous. > >Terri Clarke Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Thu, 4 Aug 2005 08:07:04 +0100 From: "Rogerson Kemlo \(ELHT\) Pathology" Subject: RE: [Histonet] slide disposal[Scanned] To: "Bonnie Whitaker" , Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698E50@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" Hire a cement mixer and get some lumps of metal or ball bearings. Put the slides and bearings in the mixer, turn on. Voila powdered glass; take to dump. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonnie Whitaker Sent: 02 August 2005 21:03 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] slide disposal[Scanned] Hi All, Can some of you give me ideas of how you handle old slide disposal? Particularly those of you in smaller, private labs. I have about 20 years worth of slides that I need to dispose of. If I have to send them with my biohazardous waste, I can, but it seems like an unnecessary expense. Currently that is what the procedure says that we do, but is it really necessary? Maybe someone out there would like to build a glass house : ) Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Thu, 4 Aug 2005 08:09:42 +0100 From: "Rogerson Kemlo \(ELHT\) Pathology" Subject: RE: [Histonet] Whole Slide Imaging[Scanned] To: "Bryan Llewellyn" , "Histonet" Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698E51@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="iso-8859-1" I'm sure you are correct but the initial bargaining went wrong when we sent an upper class twit with an attitude to the negotiating table; so I'm told. Never understood why we have less disposable earnings than the Americans AND everything costs more. === message truncated === --------------------------------- Start your day with Yahoo! - make it your home page From pruegg <@t> ihctech.net Fri Aug 5 14:04:39 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] F4/80 Ab clone CI:A3-1 not working for foam cells - why? In-Reply-To: <20050805163933.81444.qmail@web31711.mail.mud.yahoo.com> Message-ID: <200508051904.j75J4Zpf008632@chip.viawest.net> F4/80 may not pick up as many of the macrophage like cells as some of the CD68's. I really like KPMI CD68. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of GT Hebert Sent: Friday, August 05, 2005 9:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] F4/80 Ab clone CI:A3-1 not working for foam cells - why? Hello again, I just worked out my F4/80 antibody on mouse tissue. The mouse tissue, aortic arch with athrosclerotic lesions came up negative though. I would think that the foam cells (macrophages??) would light up as the do when I stain with CD68 ... but only 1 or 2 cells outside the lesion lit up. The Kupffer cells in the liver and the red pulp in the spleen (my + controls) came out great and the negative isotype controls on all samples are clear. Any idea why this may be happening? I have seen this antibody used on a number of literature papers staining foam cells in AT lesions (same clone - just different dilutions (1:25, 1:200m etc -- all paraffin) -- and they showed staining of foam cells. My tissues are all treated the same: 24 hour fixation in 4%PF, routine processed and embedded, 4 micron sections, deparaffinized ... and then IHC (optimal conditions). I'm sort of lost with this one ... I know the antibody works. Just the results are not what I expected. Any information would be greatly appreciated. Gustave Hebert Wyeth Cambridge --------------------------------- Start your day with Yahoo! - make it your home page _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From billingconsultants <@t> yahoo.com Fri Aug 5 14:19:37 2005 From: billingconsultants <@t> yahoo.com (Billing Consultants, LLC) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Multi-compartment Cassettes Message-ID: <20050805191937.23934.qmail@web54210.mail.yahoo.com> Hi everyone, Would anyone have an idea of a vendor for a multi-compartment biopsy cassette? A vendor used by one of our clients has stopped carrying them. Any direction you might provide is greatly appreciated. Thank you in advance for your assistance. Louri __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Janet.Bonner <@t> FLHOSP.ORG Fri Aug 5 14:29:50 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB435C@fh2k093.fhmis.net> OK- now I'm stepping in. Here in Florida we do get paid for days where we attend seminars/ NSH/State meetings, and many of the seminars are on campus (I'm in a hospital system) through teleconferences. External education is paid in large part by reimbursement, and, as an HTL(ASCP) with a four year degree, I'm not over-educated. And I'm Florida State Registered. Under appreciated? Not at all. Because my efforts enabled me to be where I am, I can support my life-style - and these days that's appreciation enough!! I find Histology interesting even though it may not be on a corporate pay scale. I believe being happy is paramount, and everything in this world is relative to something else. Just find your niche knowing there is always something better and other things worse. -Mom -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 8/5/2005 12:58 PM Subject: [Histonet] "Brit" Hitotech/Firestorm? Rogerson Kemlo carries on the discussion about histotechnologist training. In regards to hospital histology labs in the USA: I don't want to start a firestorm of criticism here, but I think there is a real danger that in Great Britain those technologists performing in the histology laboratories may be over qualified.A few months ago here on the Histonet, someone in the U.K. was bemoaning the lack of histology technologists in the U.K., and how short staffed they were. Was that because the bar had been set too high? Was the level of education required too much for the compensation received? Has a "closed shop" (to use a union term!) in effect, been created? Here in the United States there are hundreds of community type hospitals and smaller medical centers that have histology laboratories. In those laboratories work histotechnologists doing a fine job with a lot less education than a fully qualified histotechnologist in the U.K.. At the other end of the spectrum are histology laboratories here doing more advanced procedures where degrees and extra training are required. However, histotechnology training in the U.K. and USA are inherently different. When I lived in the U.K. years and years ago (and it may have changed now), all the hospitals were government owned, lab personnel got a day off each week with pay to attend IMLT classes, and the classes were free. To work in a hospital laboratory you had to be "State Registered". In the USA hospitals are owned by all sorts of organizations. You don't get "day release" to attend classes. If you do attend classes, money has to come from somewhere to pay for it. Further you may live miles/hours from the nearest college where you can attend classes.Since the different organisations may be "for profit" or even "Non-profit", they want to save money by not paying too higher salaries. Most often they pay what other hospitals in the area pay for the same type work. The 50 states have different requirements for working in their hospital laboratories. Hospitals have coped with these differences by making histology laboratories a separate section of the laboratory, with technicians/technologists trained just to work in that lab. I think a histotechnologist from the U.K. could easily find a position in the USA, but they might feel underappreciated and underpaid! (maybe even under challanged!) Well, that's my two bits worth! Mike Titford USA Pathology Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tracy.bergeron <@t> crl.com Fri Aug 5 15:01:54 2005 From: tracy.bergeron <@t> crl.com (tracy.bergeron@crl.com) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Multi-compartment Cassettes In-Reply-To: <20050805191937.23934.qmail@web54210.mail.yahoo.com> Message-ID: I have gotten the Evergreen Scientific multicassettes through Fisher Scientific in the past but I wasn't very fond of the plastic they are made of, the writing doesn't stay on them very well. (I have tried more different markers than I can count - and they are all about the same with these cassettes) I recently switched to getting the 6 compartment biopsy cassettes from Labstorage systems, inc. You can find them at http://www.labstore.com/ Tracy E. Bergeron, BS, HT (ASCP) Histotechnician Charles River Laboratories Wilmington, MA 978-658-6000 x 1229 "Billing Consultants, LLC" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/05/2005 03:19 PM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] Multi-compartment Cassettes Hi everyone, Would anyone have an idea of a vendor for a multi-compartment biopsy cassette? A vendor used by one of our clients has stopped carrying them. Any direction you might provide is greatly appreciated. Thank you in advance for your assistance. Louri __________________________________________________ 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 lfidgen <@t> vt.edu Fri Aug 5 15:47:28 2005 From: lfidgen <@t> vt.edu (Laura Fidgen) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Epithelial Membrane Antigen Message-ID: <6.0.0.22.0.20050805164631.0269d9d8@pop.vt.edu> Does anyone know of a reference lab that does this for dogs? Thanks, Laura L. Fidgen, MScF, BSc, HT(ASCP) Laboratory and Research Practioner VMRCVM, Histopatholgy Blacksburg, VA 24060-0443 From lrichey <@t> u.washington.edu Fri Aug 5 17:47:14 2005 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] troubles with CD4 In-Reply-To: References: Message-ID: <42F3EC72.7040302@u.washington.edu> We were having alot of problems with CD4. We use NOVACASTRA CD4 . The data sheet suggested using 0.5% hydrogen peroxide in Methanol for 10 minutes to quench. This along with 15 minutes microwave in EDTA Ph 8 has been working well in our system. Thomas Pier wrote: >Hello all, >I was wondering if anyone could help me with some problems that I'm having with staining for CD4. My control tissue for the work up is formalin fixted, paraffin embeded tonsil. I'm using Labvision's CD4 Ab-2 clone 1F6. I have diluted it anywhere from 1:10-1:50. I'm using 1% goat serum in TBS as my diluent. I've tried EDTA HIER as recomended on the data sheet. I've also tried protease XXV w/ or w/o the EDTA. I shouldn't be running into the peroxidase problem since I'm using an alkaline phosphatase detection system (Biocare's Vulcan Fast Red). I have also tried using or not using a 10% goat serum protein block. I have gotten some staining, but nowhere near as much as I should in a tonsil. Any help that you could give me would be greatly appreciated. > >Tom Pier > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From hawkmoon15 <@t> cox.net Fri Aug 5 18:12:29 2005 From: hawkmoon15 <@t> cox.net (Sarah Jones) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? References: <3E5A3F039F0BD8118B4700C00D002024043634@CKHA9> Message-ID: <004601c59a13$26ca1080$9c500644@work1> I have to remind you here, Diana, that you are speaking for yourself only with your statement about "in NA we tend to stay put". That may be for some people. However, I have found throughout my ~40 year career in Histotechnology (especially in the last half of it!!) the only way to get a decent raise is to pack up and move on. Unfortunately, here in NA our annual raises do not even keep up with the cost of living. In fact, it is down-right terrible!! I have my HTL and got my CM recently, I am proud to say. However, I haven't received any additional compensation for either, and I doubt I ever will. (HTL for 24 yrs.) Which brings up something else: The ASCP used to be the American Society of Clinical Pathologists. It is now the American Society of Clinical Pathology. What changed? I'm not sure. Does anyone know, except the 'insiders', that is? I do have first-hand knowledge of what used to be. The Pathologists ran it, and they had a lot to do with where Histotechs sat -- low on the rung of the Pathology Laboratory ladder for YEARS. The Registry was in my home town in Muncie, Indiana, and my Mother was the secretary to the Registrar for many years. (This was all before it was centralized in Chicago. Prior to the relocation to Chicago, the Registry was in the town where the Chairman of the Board was located. L. Montgomery, M.D., was the Chairman of the Board for ~30 years that I know of, and thus was the Registry located in Muncie.) There are other things I could tell you; but I was sworn to secrecy years ago, and I will honor that pledge. Suffice it to say, the Pathologists wanted to keep the salaries of the Histotechs down, and did everything in their power to do so. 'Nuff said! When I started into the field of Histotechnology at the University of Chicago back in 1965, the Techies associated with the ASCP were trying to change the requirements for Histotechs, but to no avail. Not until January of 2005 did the change they were attempting to accomplish way back then prevail. I didn't really understand what was happening then, but today I do. There are MANY places here in the USA (where there is a severe shortage of Histotechs) where they will still hire techs that are not licensed. In fact, there are places where a licensed Histologic Technician OR a licensed Histotechnologist are black-listed by the other techs--most likely in fear that their own incompetence might be discovered! The cost of living in certain areas of the country are no longer taken into consideration in the way they were in the past when it comes to hiring techs. That also saddens me. How is it that they think I can live in the most expensive county in California that pays the lowest wages of any county in California?? That is craziness in my book!!!! Yet they wonder why they cannot find good Histotechs here in this county! GO FIGURE!! Anyway, I could go on and on, but I won't. Hope this sheds some light on some things for some folks. Please address any personal questions to: hawkmoon15@cox.net Sarah ----- Original Message ----- From: "Diana McCaig" To: Sent: Friday, August 05, 2005 10:41 AM Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? > > What are you all using to compare the wage scales. Biomedical scientists do > not get paid as well as we do in North America if you factor in their cost > of living. I understand there is a recent trend to increase the wages in > order to keep them on board as there is a lot of migration to various labs > throughout a career where in NA we tend to stay put. > Diana McCaig, MLT > > > -----Original Message----- > From: Larry Woody [SMTP:slappycraw@yahoo.com] > Sent: Friday, August 05, 2005 1:35 PM > To: mtitford@aol.com; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > You may want to try some of the big biotech companies where there is > a need for people with more skills in the lab and the compensation can be > better than a medical lab. > > mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion > about histotechnologist training. > > In regards to hospital histology labs in the USA: > > I don't want to start a firestorm of criticism here, but I think > there is a real danger that in Great Britain those technologists performing > in the histology laboratories may be over qualified.A few months ago here on > the Histonet, someone in the U.K. was bemoaning the lack of histology > technologists in the U.K., and how short staffed they were. Was that because > the bar had been set too high? Was the level of education required too much > for the compensation received? Has a "closed shop" (to use a union term!) in > effect, been created? > Here in the United States there are hundreds of community type > hospitals and smaller medical centers that have histology laboratories. In > those laboratories work histotechnologists doing a fine job with a lot less > education than a fully qualified histotechnologist in the U.K.. At the other > end of the spectrum are histology laboratories here doing more advanced > procedures where degrees and extra training are required. > However, histotechnology training in the U.K. and USA are inherently > different. When I lived in the U.K. years and years ago (and it may have > changed now), all the hospitals were government owned, lab personnel got a > day off each week with pay to attend IMLT classes, and the classes were > free. To work in a hospital laboratory you had to be "State Registered". > In the USA hospitals are owned by all sorts of organizations. You > don't get "day release" to attend classes. If you do attend classes, money > has to come from somewhere to pay for it. Further you may live miles/hours > from the nearest college where you can attend classes.Since the different > organisations may be "for profit" or even "Non-profit", they want to save > money by not paying too higher salaries. Most often they pay what other > hospitals in the area pay for the same type work. > The 50 states have different requirements for working in their > hospital laboratories. > Hospitals have coped with these differences by making histology > laboratories a separate section of the laboratory, with > technicians/technologists trained just to work in that lab. > I think a histotechnologist from the U.K. could easily find a > position in the USA, but they might feel underappreciated and underpaid! > (maybe even under challanged!) > Well, that's my two bits worth! > > Mike Titford > USA Pathology > Mobile AL USA > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > --------------------------------- > Start your day with Yahoo! - make it your home page > _______________________________________________ > Histonet mailing list > Histonet@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 clarke.ian <@t> virgin.net Sat Aug 6 06:48:58 2005 From: clarke.ian <@t> virgin.net (Ian Clarke) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] RE:deterioration of paraplast wax blocks In-Reply-To: <004601c59a13$26ca1080$9c500644@work1> Message-ID: Hi All Some of our stored blocks are detoriorating due to a ? fungus which is eating them. We have asked the micro people to see if they could identify it but they have been unable to make it grow.Has anyone had a similar problem and if so how did you rectify it. Where you able to classify the organism. ian clarke Cellular Pathology Department Craigavon Area Hospital From lubbockcat <@t> hotmail.com Sat Aug 6 07:33:58 2005 From: lubbockcat <@t> hotmail.com (Terry Murphy) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? In-Reply-To: <004601c59a13$26ca1080$9c500644@work1> Message-ID: HI everyone I feel the urge to join this discussion. I got into the field of histology 15 years ago and enjoyed the work and the concepts. I wanted to move up in the field ao I earned my HTL (ASCP) instead of the HT(ASCP) since I have a Bachelor's degree in Biology. That didn't get me any more respect or money. Still trying to improve my situation I earn a Master's degree in Health Administration and I once again my employment has not improved. As a matter of fact my employment situation has gotten worse since I earned a Master's degree. I had acheived a position as a supervisor only to be "run out of the lab" by an arrogant PA and, I can only assume, seasoned histotechs that fear change. Any way now I have been working as a traveling histotech. The pay is better than if I had a permanent job. The facility I am at now recently hired a person to be trained as a tech. This new employee has a GED not a high school dipolma and he has no background in in science or medicine. He is doing good given his background but I find it kind of a insult to educated techs like myself that that someone without education or experience has been hired to do the same job that us techs have studied extensively. Now I am done traveling and am trying to get a histotech job close to my home town so I can live in my house and more importantly live with my wife. I have applied for tech positions and either haven't heard from the facility except for a rejection letter. When I can get a hold of a hireing supervisor I have been told that "this is a entry level position and it doesn't pay well", "you are overqualified", and, this really surprised me, "your skill set will not go well in our laboratory". Someone on this board mentioned about being "black listed" I feel that I have now entered that list. Most likely I have more education than the people that I would be reporting to and they probably feel threatened by me. Someone suggested that I "dumb down" my resume and exclude some education and my publication but I do not like the idea of excluding accomplishments that I am proud of. Lastly, as for wages, histotechs are grossly underpaid while Pathologist Assistants are grossly overpaid!!!! Thanks for letting me vent. Sincerely, Another fustrated histotech. >From: "Sarah Jones" >To: >Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? >Date: Fri, 5 Aug 2005 16:12:29 -0700 > >I have to remind you here, Diana, that you are speaking for yourself only >with your statement about "in NA we tend to stay put". That may be for >some >people. However, I have found throughout my ~40 year career in >Histotechnology (especially in the last half of it!!) the only way to get a >decent raise is to pack up and move on. Unfortunately, here in NA our >annual raises do not even keep up with the cost of living. In fact, it is >down-right terrible!! I have my HTL and got my CM recently, I am proud to >say. However, I haven't received any additional compensation for either, >and I doubt I ever will. (HTL for 24 yrs.) > >Which brings up something else: The ASCP used to be the American Society >of >Clinical Pathologists. It is now the American Society of Clinical >Pathology. What changed? I'm not sure. Does anyone know, except the >'insiders', that is? I do have first-hand knowledge of what used to be. >The Pathologists ran it, and they had a lot to do with where Histotechs >sat -- low on the rung of the Pathology Laboratory ladder for YEARS. The >Registry was in my home town in Muncie, Indiana, and my Mother was the >secretary to the Registrar for many years. (This was all before it was >centralized in Chicago. Prior to the relocation to Chicago, the Registry >was in the town where the Chairman of the Board was located. L. >Montgomery, >M.D., was the Chairman of the Board for ~30 years that I know of, and thus >was the Registry located in Muncie.) There are other things I could tell >you; but I was sworn to secrecy years ago, and I will honor that pledge. >Suffice it to say, the Pathologists wanted to keep the salaries of the >Histotechs down, and did everything in their power to do so. 'Nuff said! > >When I started into the field of Histotechnology at the University of >Chicago back in 1965, the Techies associated with the ASCP were trying to >change the requirements for Histotechs, but to no avail. Not until January >of 2005 did the change they were attempting to accomplish way back then >prevail. I didn't really understand what was happening then, but today I >do. There are MANY places here in the USA (where there is a severe >shortage >of Histotechs) where they will still hire techs that are not licensed. In >fact, there are places where a licensed Histologic Technician OR a licensed >Histotechnologist are black-listed by the other techs--most likely in fear >that their own incompetence might be discovered! > >The cost of living in certain areas of the country are no longer taken into >consideration in the way they were in the past when it comes to hiring >techs. That also saddens me. How is it that they think I can live in the >most expensive county in California that pays the lowest wages of any >county >in California?? That is craziness in my book!!!! Yet they wonder why they >cannot find good Histotechs here in this county! GO FIGURE!! > >Anyway, I could go on and on, but I won't. Hope this sheds some light on >some things for some folks. Please address any personal questions to: >hawkmoon15@cox.net > >Sarah > > >----- Original Message ----- >From: "Diana McCaig" >To: >Sent: Friday, August 05, 2005 10:41 AM >Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > What are you all using to compare the wage scales. Biomedical >scientists >do > > not get paid as well as we do in North America if you factor in their >cost > > of living. I understand there is a recent trend to increase the wages >in > > order to keep them on board as there is a lot of migration to various >labs > > throughout a career where in NA we tend to stay put. > > Diana McCaig, MLT > > > > > > -----Original Message----- > > From: Larry Woody [SMTP:slappycraw@yahoo.com] > > Sent: Friday, August 05, 2005 1:35 PM > > To: mtitford@aol.com; histonet@lists.utsouthwestern.edu > > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > > > You may want to try some of the big biotech companies where there is > > a need for people with more skills in the lab and the compensation can >be > > better than a medical lab. > > > > mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion > > about histotechnologist training. > > > > In regards to hospital histology labs in the USA: > > > > I don't want to start a firestorm of criticism here, but I think > > there is a real danger that in Great Britain those technologists >performing > > in the histology laboratories may be over qualified.A few months ago >here >on > > the Histonet, someone in the U.K. was bemoaning the lack of histology > > technologists in the U.K., and how short staffed they were. Was that >because > > the bar had been set too high? Was the level of education required too >much > > for the compensation received? Has a "closed shop" (to use a union >term!) >in > > effect, been created? > > Here in the United States there are hundreds of community type > > hospitals and smaller medical centers that have histology laboratories. >In > > those laboratories work histotechnologists doing a fine job with a lot >less > > education than a fully qualified histotechnologist in the U.K.. At the >other > > end of the spectrum are histology laboratories here doing more advanced > > procedures where degrees and extra training are required. > > However, histotechnology training in the U.K. and USA are inherently > > different. When I lived in the U.K. years and years ago (and it may have > > changed now), all the hospitals were government owned, lab personnel got >a > > day off each week with pay to attend IMLT classes, and the classes were > > free. To work in a hospital laboratory you had to be "State Registered". > > In the USA hospitals are owned by all sorts of organizations. You > > don't get "day release" to attend classes. If you do attend classes, >money > > has to come from somewhere to pay for it. Further you may live >miles/hours > > from the nearest college where you can attend classes.Since the >different > > organisations may be "for profit" or even "Non-profit", they want to >save > > money by not paying too higher salaries. Most often they pay what other > > hospitals in the area pay for the same type work. > > The 50 states have different requirements for working in their > > hospital laboratories. > > Hospitals have coped with these differences by making histology > > laboratories a separate section of the laboratory, with > > technicians/technologists trained just to work in that lab. > > I think a histotechnologist from the U.K. could easily find a > > position in the USA, but they might feel underappreciated and underpaid! > > (maybe even under challanged!) > > Well, that's my two bits worth! > > > > Mike Titford > > USA Pathology > > Mobile AL USA > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > --------------------------------- > > Start your day with Yahoo! - make it your home page > > _______________________________________________ > > Histonet mailing list > > Histonet@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 _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From Adesupod <@t> aol.com Sat Aug 6 10:30:23 2005 From: Adesupod <@t> aol.com (Adesupod@aol.com) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: <145.4ad2f20c.3026318f@aol.com> Hi Everyone, I think the issue of allowing people with G.E.D to do on-the job training in Histotechnology is demeaning and really suppressing the status of Histotechnologists in the United States. More-over I did not blame the Pathologist Assistants, Cytotechnologists and Med. Techs that feels that they are more superior and more educated in comparison to the Histotechs. In United Kingdom, Australia, Canada, New Zealand and Nigeria, the trainig is the same for all the Medical Lab.Scientists/ Clinical Lab.Scientists. The Histology Technologists in all those countries are college graduates( 4 to 5 years degree program), and they are registered as Medical Technologists/Clinical Lab.Scientists. I think the issue of low pay for the Histotechs in the United States, will continue for a very long time, until when the minimum entry into the profession is being raised to BS degree. Actually, I am thinking of sitting for my MT(ASCP) certification test, and change from Histotech to Med.Tech.( thank God I have the qualification). From another frustrated Histotech. From gu.lang <@t> gmx.at Sat Aug 6 12:56:35 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] tween detergens Message-ID: Hi all, We are going to run FISH on our ventana benchmark. In the last step we have to wash the slides to remove the liquid cover slip. Until now we use a green dish-washing product for the ihc, but that doesn't fit with the FISH because of autofluorescence. Can anyone give me a hint, if Tween 20 in ?% solution in water is okay? Or any other well working detergens? Our Ventana-Lady is also looking for a colourless dish-washer, but with no results yet. Thank you in advance Gudrun Lang Linz, Austria From beingmary53 <@t> sbcglobal.net Sat Aug 6 13:16:28 2005 From: beingmary53 <@t> sbcglobal.net (MARY JOHNSON) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] stain Message-ID: <20050806181628.33904.qmail@web81610.mail.yahoo.com> Hi Herbert, have you tried the Movat Pentachrome stain, if you need the procedure sent me your email and I will sent you the procedure. Mary From Charles.Embrey <@t> carle.com Sat Aug 6 13:55:20 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: Hi Terry, Please know that some Pathologists' Assistants belong to this list as well and I really don't like someone not in my shoes telling me that I am overpaid. I spent 20 years in the Air Force and busted my @#%@# as a Histotech and Histology Supervisor for less than underpaid civilian histotechs make. But I really wasn't in the military for the money. I finished my education and joined the AAPA, passing my Fellowship exam after retiring from the Air Force. I am here at work on a Saturday, after already putting in a 40 hour week so that I can finish up some paperwork I couldn't complete during the week because of the 500+ cases I grossed and the two cancer conferences I needed to attend. I do agree that histotechs are behind on the pay scale but don't let your "sour grapes" cause you to spout off about a profession you don't work in. Education doesn't always equate to salary. I personally know some histotechs with only high school education that can work circles around BS educated registered HTL's. They have a fully working knowledge of histology theory, do a great job and have my full respect. To me you sound like someone that "toots his own horn" a lot and probably carries mini copies of your diplomas in you wallet. I am sorry that you met an "arrogant PA" but we are not all like that. I am normally a pretty easy going, laid back southern boy except when someone spouts of generalizations that are untrue out of self pity over his own situation. Charles Embrey, HT, PA(ASCP) Histology Manager Carle Clinic Illinois -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Terry Murphy Sent: Saturday, August 06, 2005 7:34 AM To: hawkmoon15@cox.net; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? HI everyone I feel the urge to join this discussion. I got into the field of histology 15 years ago and enjoyed the work and the concepts. I wanted to move up in the field ao I earned my HTL (ASCP) instead of the HT(ASCP) since I have a Bachelor's degree in Biology. That didn't get me any more respect or money. Still trying to improve my situation I earn a Master's degree in Health Administration and I once again my employment has not improved. As a matter of fact my employment situation has gotten worse since I earned a Master's degree. I had acheived a position as a supervisor only to be "run out of the lab" by an arrogant PA and, I can only assume, seasoned histotechs that fear change. Any way now I have been working as a traveling histotech. The pay is better than if I had a permanent job. The facility I am at now recently hired a person to be trained as a tech. This new employee has a GED not a high school dipolma and he has no background in in science or medicine. He is doing good given his background but I find it kind of a insult to educated techs like myself that that someone without education or experience has been hired to do the same job that us techs have studied extensively. Now I am done traveling and am trying to get a histotech job close to my home town so I can live in my house and more importantly live with my wife. I have applied for tech positions and either haven't heard from the facility except for a rejection letter. When I can get a hold of a hireing supervisor I have been told that "this is a entry level position and it doesn't pay well", "you are overqualified", and, this really surprised me, "your skill set will not go well in our laboratory". Someone on this board mentioned about being "black listed" I feel that I have now entered that list. Most likely I have more education than the people that I would be reporting to and they probably feel threatened by me. Someone suggested that I "dumb down" my resume and exclude some education and my publication but I do not like the idea of excluding accomplishments that I am proud of. Lastly, as for wages, histotechs are grossly underpaid while Pathologist Assistants are grossly overpaid!!!! Thanks for letting me vent. Sincerely, Another fustrated histotech. >From: "Sarah Jones" >To: >Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? >Date: Fri, 5 Aug 2005 16:12:29 -0700 > >I have to remind you here, Diana, that you are speaking for yourself only >with your statement about "in NA we tend to stay put". That may be for >some >people. However, I have found throughout my ~40 year career in >Histotechnology (especially in the last half of it!!) the only way to get a >decent raise is to pack up and move on. Unfortunately, here in NA our >annual raises do not even keep up with the cost of living. In fact, it is >down-right terrible!! I have my HTL and got my CM recently, I am proud to >say. However, I haven't received any additional compensation for either, >and I doubt I ever will. (HTL for 24 yrs.) > >Which brings up something else: The ASCP used to be the American Society >of >Clinical Pathologists. It is now the American Society of Clinical >Pathology. What changed? I'm not sure. Does anyone know, except the >'insiders', that is? I do have first-hand knowledge of what used to be. >The Pathologists ran it, and they had a lot to do with where Histotechs >sat -- low on the rung of the Pathology Laboratory ladder for YEARS. The >Registry was in my home town in Muncie, Indiana, and my Mother was the >secretary to the Registrar for many years. (This was all before it was >centralized in Chicago. Prior to the relocation to Chicago, the Registry >was in the town where the Chairman of the Board was located. L. >Montgomery, >M.D., was the Chairman of the Board for ~30 years that I know of, and thus >was the Registry located in Muncie.) There are other things I could tell >you; but I was sworn to secrecy years ago, and I will honor that pledge. >Suffice it to say, the Pathologists wanted to keep the salaries of the >Histotechs down, and did everything in their power to do so. 'Nuff said! > >When I started into the field of Histotechnology at the University of >Chicago back in 1965, the Techies associated with the ASCP were trying to >change the requirements for Histotechs, but to no avail. Not until January >of 2005 did the change they were attempting to accomplish way back then >prevail. I didn't really understand what was happening then, but today I >do. There are MANY places here in the USA (where there is a severe >shortage >of Histotechs) where they will still hire techs that are not licensed. In >fact, there are places where a licensed Histologic Technician OR a licensed >Histotechnologist are black-listed by the other techs--most likely in fear >that their own incompetence might be discovered! > >The cost of living in certain areas of the country are no longer taken into >consideration in the way they were in the past when it comes to hiring >techs. That also saddens me. How is it that they think I can live in the >most expensive county in California that pays the lowest wages of any >county >in California?? That is craziness in my book!!!! Yet they wonder why they >cannot find good Histotechs here in this county! GO FIGURE!! > >Anyway, I could go on and on, but I won't. Hope this sheds some light on >some things for some folks. Please address any personal questions to: >hawkmoon15@cox.net > >Sarah > > >----- Original Message ----- >From: "Diana McCaig" >To: >Sent: Friday, August 05, 2005 10:41 AM >Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > What are you all using to compare the wage scales. Biomedical >scientists >do > > not get paid as well as we do in North America if you factor in their >cost > > of living. I understand there is a recent trend to increase the wages >in > > order to keep them on board as there is a lot of migration to various >labs > > throughout a career where in NA we tend to stay put. > > Diana McCaig, MLT > > > > > > -----Original Message----- > > From: Larry Woody [SMTP:slappycraw@yahoo.com] > > Sent: Friday, August 05, 2005 1:35 PM > > To: mtitford@aol.com; histonet@lists.utsouthwestern.edu > > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > > > You may want to try some of the big biotech companies where there is > > a need for people with more skills in the lab and the compensation can >be > > better than a medical lab. > > > > mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion > > about histotechnologist training. > > > > In regards to hospital histology labs in the USA: > > > > I don't want to start a firestorm of criticism here, but I think > > there is a real danger that in Great Britain those technologists >performing > > in the histology laboratories may be over qualified.A few months ago >here >on > > the Histonet, someone in the U.K. was bemoaning the lack of histology > > technologists in the U.K., and how short staffed they were. Was that >because > > the bar had been set too high? Was the level of education required too >much > > for the compensation received? Has a "closed shop" (to use a union >term!) >in > > effect, been created? > > Here in the United States there are hundreds of community type > > hospitals and smaller medical centers that have histology laboratories. >In > > those laboratories work histotechnologists doing a fine job with a lot >less > > education than a fully qualified histotechnologist in the U.K.. At the >other > > end of the spectrum are histology laboratories here doing more advanced > > procedures where degrees and extra training are required. > > However, histotechnology training in the U.K. and USA are inherently > > different. When I lived in the U.K. years and years ago (and it may have > > changed now), all the hospitals were government owned, lab personnel got >a > > day off each week with pay to attend IMLT classes, and the classes were > > free. To work in a hospital laboratory you had to be "State Registered". > > In the USA hospitals are owned by all sorts of organizations. You > > don't get "day release" to attend classes. If you do attend classes, >money > > has to come from somewhere to pay for it. Further you may live >miles/hours > > from the nearest college where you can attend classes.Since the >different > > organisations may be "for profit" or even "Non-profit", they want to >save > > money by not paying too higher salaries. Most often they pay what other > > hospitals in the area pay for the same type work. > > The 50 states have different requirements for working in their > > hospital laboratories. > > Hospitals have coped with these differences by making histology > > laboratories a separate section of the laboratory, with > > technicians/technologists trained just to work in that lab. > > I think a histotechnologist from the U.K. could easily find a > > position in the USA, but they might feel underappreciated and underpaid! > > (maybe even under challanged!) > > Well, that's my two bits worth! > > > > Mike Titford > > USA Pathology > > Mobile AL USA > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > --------------------------------- > > Start your day with Yahoo! - make it your home page > > _______________________________________________ > > Histonet mailing list > > Histonet@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 _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From daufoi <@t> msn.com Sat Aug 6 18:44:42 2005 From: daufoi <@t> msn.com (Wisam Barkho) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] fixation via perfusion Message-ID: Hello, We are doing stereotaxic surgeries on rats in my lab. After 2 weeks, we fixate the brain via perfusion using 4% PFA (after vascular rinse) and then immerse the brains in 4% PFA overnight. My question is where can I find information on pump flow rate and PFA volume to use during fixation? Thanks. Wisam Barkho daufoi@msn.com From amosbrooks <@t> earthlink.net Sat Aug 6 21:34:31 2005 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] Re: troubles with CD4 In-Reply-To: <200508051300.1e15yS4hi3Nl34k1@mx-avoceta.atl.sa.earthlink.net> References: <200508051300.1e15yS4hi3Nl34k1@mx-avoceta.atl.sa.earthlink.net> Message-ID: <42F57337.3000605@earthlink.net> Tom, What is the pH of the retrieval solution that you are using? In our experience CD4 tends to work best with a high pH retrieval solution (like Dako's high pH retrieval solution or Biocare Medical's Borg Decloaker). We use Sigma's Impress for detection of this because of it being a polymer which does not use avidin/ biotin it avoids the endogenous biotin background that high pH retrieval tends to increase. Just some suggestions to try, Amos Brooks Message: 15 Date: Fri, 05 Aug 2005 10:47:22 -0500 From: "Thomas Pier" Subject: [Histonet] troubles with CD4 To: Message-ID: Content-Type: text/plain; charset=US-ASCII Hello all, I was wondering if anyone could help me with some problems that I'm having with staining for CD4. My control tissue for the work up is formalin fixted, paraffin embeded tonsil. I'm using Labvision's CD4 Ab-2 clone 1F6. I have diluted it anywhere from 1:10-1:50. I'm using 1% goat serum in TBS as my diluent. I've tried EDTA HIER as recomended on the data sheet. I've also tried protease XXV w/ or w/o the EDTA. I shouldn't be running into the peroxidase problem since I'm using an alkaline phosphatase detection system (Biocare's Vulcan Fast Red). I have also tried using or not using a 10% goat serum protein block. I have gotten some staining, but nowhere near as much as I should in a tonsil. Any help that you could give me would be greatly appreciated. Tom Pier From sasha_sashimi <@t> yahoo.com Sun Aug 7 02:39:35 2005 From: sasha_sashimi <@t> yahoo.com (Sasha Manhattan) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] help: Heidanhein's azan trichrome Message-ID: <20050807073935.7707.qmail@web31201.mail.mud.yahoo.com> Hello colleagues, I am a student at the University of Hong Kong. I am trying Heidanhein's azan staining to quantitate the extent of fibrosis in mouse liver. When I tried it for the first time, there was a blue colour in the tissues showing collagen deposition, which is what I expected, but the protocol I used does not include aniline. I only used 95% alcohol to extract azaocarmine G from collagen because our lab do not have aniline at the moment. but since i got the blue colour, i think it was ok. but now that I repeat it with the same tissue, i dont see any blue colour. I thought maybe the distilled water rinse washed out the water-soluble aniline blue, but i repeated without washing in distilled water (just put in 95% alcohol and then absolute alcohol and then xylene clear), but there's still no blue colour. Could you tell me what could be wrong? This is my protocol: 1.deparaffination, hydration through alcohols. 2. Put slides into azocarmine G solution at 50 degrees celsius for 1 hour. 3. Rinse quickly with dH2O. 4. Place into 95% ethanol. 5. Place into 1% acetic acid in 95% ethanol. 6. Rinse briefly in dH2O. 7. Place into 5% phosphotungstic acid for 2 hrs. 8. Place into aniline blue-orange G solution for 2 hrs. 9. Rinse very quickly with dH2O, dehydrate with 95% and 100% ethanol(just dip), 3 changes of xylene each of 3 minutes. mount. I would be very grateful if you could just tell me what might have gone wrong. There is no one for me to ask in the lab because no one does histology here, and I am really nervous and frustrated. Thank you very much for your time and attention. Yours sincerely, Mimi Kwun-nok Man --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour From Kemlo.Rogerson <@t> elht.nhs.uk Sat Aug 6 03:01:17 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4D2@elht-exch1.xelht.nhs.uk> Thank you for that; what you say is very interesting and I agree with a lot of it. Interestingly, whether it's because it's Holiday time, no-one from the UK has answered my implied question. Are we overqualified in the UK to be 'HistoTechs'; even more contentious, what is the role of the Medical Laboratory assistant (Higher or even Associate Practitioner)? Are they US Histotechs in disguise? Do we (Biomedical Scientists) therefore, not exist in the US, and why not? Kemlo Rogerson MSc MIBiol CBiol CSci DMS FIBMS -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of mtitford@aol.com Sent: 05 August 2005 17:59 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] "Brit" Hitotech/Firestorm? Rogerson Kemlo carries on the discussion about histotechnologist training. In regards to hospital histology labs in the USA: I don't want to start a firestorm of criticism here, but I think there is a real danger that in Great Britain those technologists performing in the histology laboratories may be over qualified.A few months ago here on the Histonet, someone in the U.K. was bemoaning the lack of histology technologists in the U.K., and how short staffed they were. Was that because the bar had been set too high? Was the level of education required too much for the compensation received? Has a "closed shop" (to use a union term!) in effect, been created? Here in the United States there are hundreds of community type hospitals and smaller medical centers that have histology laboratories. In those laboratories work histotechnologists doing a fine job with a lot less education than a fully qualified histotechnologist in the U.K.. At the other end of the spectrum are histology laboratories here doing more advanced procedures where degrees and extra training are required. However, histotechnology training in the U.K. and USA are inherently different. When I lived in the U.K. years and years ago (and it may have changed now), all the hospitals were government owned, lab personnel got a day off each week with pay to attend IMLT classes, and the classes were free. To work in a hospital laboratory you had to be "State Registered". In the USA hospitals are owned by all sorts of organizations. You don't get "day release" to attend classes. If you do attend classes, money has to come from somewhere to pay for it. Further you may live miles/hours from the nearest college where you can attend classes.Since the different organisations may be "for profit" or even "Non-profit", they want to save money by not paying too higher salaries. Most often they pay what other hospitals in the area pay for the same type work. The 50 states have different requirements for working in their hospital laboratories. Hospitals have coped with these differences by making histology laboratories a separate section of the laboratory, with technicians/technologists trained just to work in that lab. I think a histotechnologist from the U.K. could easily find a position in the USA, but they might feel underappreciated and underpaid! (maybe even under challanged!) Well, that's my two bits worth! Mike Titford USA Pathology Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Sun Aug 7 23:59:47 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] tween detergens References: <0IKT007S1BCK4B20@chico.mail.uwo.pri> <42F6E451.8A1E1D8D@uwo.ca> Message-ID: <42F6E6C3.6B706C60@uwo.ca> John Kiernan wrote: > > Why use dish-washing liquid, of unknown composition, in > an expensive procedure in a laboratory? The pure chemicals > are not expensive and are available from regular suppliers. > For example, Tween 20 (an old trade-name for > polyoxyethylene sorbitan monolaurate) costs about US$25 > for 100 ml of syrupy liquid - enough to last a long, long > time. Even if the price in Europe is 4 times as high, > this is nothing when compared with the cost of the > nucleic acid probes and other expendable supplies > needed for in situ hybridization. > > If your technique calls for a detergent, use the one > in the published account of the method, at the concentration > that the author found to be effective. If you do anything > else you are wasting time and money. Never try to improve > on a technique before you have tried the original! > > Someone worked hard to optimize the method, and got it > published in a peer-reviewed journal. It may well be possible > to make improvements, but the starting point must be > repeating a procedure that has worked for others. You must > faithfully follow the original published procedure, using the > same kind of cells or tissues, and including proper positive > and negative controls. If you fail to get even a hint of a > meaningful result, despite doing everything by the book, > and the appearances of the controls don't help, you may be > missing out on some simple consideration that is not a major > part of the method. > > Your mention of "green dish-washing liquid" in the same > sentence as "autofluorescence" indicates that you (or your > boss) may not fully understand the ways words are used by > people who do fluorescence microscopy. > > Autofluorescence means "self-fluorescence"; it originates > in the object being examined. Fluorescence derived from a > staining method is not autofluorescence. Unwanted fluorescence > from a coloured dishwashing liquid is optical pollution. > > Bottom line. 1. Got to the library. 2. Follow the procedure. > > John Kiernan > Anatomy Dept, UWO > London, Canada > ------------------------------------------------ > Gudrun Lang wrote: > > > > Hi all, > > > > We are going to run FISH on our ventana benchmark. In the last step we have > > to wash the slides to remove the liquid cover slip. Until now we use a green > > dish-washing product for the ihc, but that doesn't fit with the FISH because > > of autofluorescence. > > > > Can anyone give me a hint, if Tween 20 in ?% solution in water is okay? Or > > any other well working detergens? > > > > Our Ventana-Lady is also looking for a colourless dish-washer, but with no > > results yet. > > > > > > > > Thank you in advance > > > > Gudrun Lang > > > > Linz, Austria > > *** Gudrun Lang: if you identify yourself and your > laboratory more clearly, you may attract expert > attention. > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ From jkiernan <@t> uwo.ca Mon Aug 8 00:00:37 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] tween detergens References: <0IKT007S1BCK4B20@chico.mail.uwo.pri> <42F6E451.8A1E1D8D@uwo.ca> Message-ID: <42F6E6F5.2F15967D@uwo.ca> John Kiernan wrote: > > Why use dish-washing liquid, of unknown composition, in > an expensive procedure in a laboratory? The pure chemicals > are not expensive and are available from regular suppliers. > For example, Tween 20 (an old trade-name for > polyoxyethylene sorbitan monolaurate) costs about US$25 > for 100 ml of syrupy liquid - enough to last a long, long > time. Even if the price in Europe is 4 times as high, > this is nothing when compared with the cost of the > nucleic acid probes and other expendable supplies > needed for in situ hybridization. > > If your technique calls for a detergent, use the one > in the published account of the method, at the concentration > that the author found to be effective. If you do anything > else you are wasting time and money. Never try to improve > on a technique before you have tried the original! > > Someone worked hard to optimize the method, and got it > published in a peer-reviewed journal. It may well be possible > to make improvements, but the starting point must be > repeating a procedure that has worked for others. You must > faithfully follow the original published procedure, using the > same kind of cells or tissues, and including proper positive > and negative controls. If you fail to get even a hint of a > meaningful result, despite doing everything by the book, > and the appearances of the controls don't help, you may be > missing out on some simple consideration that is not a major > part of the method. > > Your mention of "green dish-washing liquid" in the same > sentence as "autofluorescence" indicates that you (or your > boss) may not fully understand the ways words are used by > people who do fluorescence microscopy. > > Autofluorescence means "self-fluorescence"; it originates > in the object being examined. Fluorescence derived from a > staining method is not autofluorescence. Unwanted fluorescence > from a coloured dishwashing liquid is optical pollution. > > Bottom line. 1. Got to the library. 2. Follow the procedure. > > John Kiernan > Anatomy Dept, UWO > London, Canada > ------------------------------------------------ > Gudrun Lang wrote: > > > > Hi all, > > > > We are going to run FISH on our ventana benchmark. In the last step we have > > to wash the slides to remove the liquid cover slip. Until now we use a green > > dish-washing product for the ihc, but that doesn't fit with the FISH because > > of autofluorescence. > > > > Can anyone give me a hint, if Tween 20 in ?% solution in water is okay? Or > > any other well working detergens? > > > > Our Ventana-Lady is also looking for a colourless dish-washer, but with no > > results yet. > > > > > > > > Thank you in advance > > > > Gudrun Lang > > > > Linz, Austria > > *** Gudrun Lang: if you identify yourself and your > laboratory more clearly, you may attract expert > attention. > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ From Kemlo.Rogerson <@t> elht.nhs.uk Mon Aug 8 03:02:11 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698E71@elht-exch1.xelht.nhs.uk> Actually you are incorrect, we are called Biomedical Scientists and there are many ways of entering the profession all concluding, I admit, with a BSc (Hons). Clinical Scientists are different than BMS but why alludes me! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Adesupod@aol.com Sent: 06 August 2005 16:30 To: lubbockcat@hotmail.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? Hi Everyone, I think the issue of allowing people with G.E.D to do on-the job training in Histotechnology is demeaning and really suppressing the status of Histotechnologists in the United States. More-over I did not blame the Pathologist Assistants, Cytotechnologists and Med. Techs that feels that they are more superior and more educated in comparison to the Histotechs. In United Kingdom, Australia, Canada, New Zealand and Nigeria, the trainig is the same for all the Medical Lab.Scientists/ Clinical Lab.Scientists. The Histology Technologists in all those countries are college graduates( 4 to 5 years degree program), and they are registered as Medical Technologists/Clinical Lab.Scientists. I think the issue of low pay for the Histotechs in the United States, will continue for a very long time, until when the minimum entry into the profession is being raised to BS degree. Actually, I am thinking of sitting for my MT(ASCP) certification test, and change from Histotech to Med.Tech.( thank God I have the qualification). From another frustrated Histotech. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Mon Aug 8 03:13:47 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4D3@elht-exch1.xelht.nhs.uk> To give you an idea over pay. Band 5 (equates to a newly qualified BMS) ?18,698 to ?24,198; Band 6 (equates to an experienced BMS) ?21,448 to ?30,247; Band 7 (equates to a BMS with an MSc and 'specialised') ?25,628 to ?35,527; Band 8A (Professional Manager Level 1) ?31,127 to ?41,246; Band 8b (Professional Manager Lev 2) ?35,527 to ?49,496; Band 8c (Professional Manager Lev 3/ Advanced Practitioner) ?41,246 to ?59,395; Band 8D (Head of Service?) ?49,496 to ?71,494 and finally Band 9 (Pathology Director) ?59,395 to ?86,240. There are few 8D 's and 9's around; they are fabled creatures heard of but never seen. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Diana McCaig Sent: 05 August 2005 18:41 To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? What are you all using to compare the wage scales. Biomedical scientists do not get paid as well as we do in North America if you factor in their cost of living. I understand there is a recent trend to increase the wages in order to keep them on board as there is a lot of migration to various labs throughout a career where in NA we tend to stay put. Diana McCaig, MLT -----Original Message----- From: Larry Woody [SMTP:slappycraw@yahoo.com] Sent: Friday, August 05, 2005 1:35 PM To: mtitford@aol.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? You may want to try some of the big biotech companies where there is a need for people with more skills in the lab and the compensation can be better than a medical lab. mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion about histotechnologist training. In regards to hospital histology labs in the USA: I don't want to start a firestorm of criticism here, but I think there is a real danger that in Great Britain those technologists performing in the histology laboratories may be over qualified.A few months ago here on the Histonet, someone in the U.K. was bemoaning the lack of histology technologists in the U.K., and how short staffed they were. Was that because the bar had been set too high? Was the level of education required too much for the compensation received? Has a "closed shop" (to use a union term!) in effect, been created? Here in the United States there are hundreds of community type hospitals and smaller medical centers that have histology laboratories. In those laboratories work histotechnologists doing a fine job with a lot less education than a fully qualified histotechnologist in the U.K.. At the other end of the spectrum are histology laboratories here doing more advanced procedures where degrees and extra training are required. However, histotechnology training in the U.K. and USA are inherently different. When I lived in the U.K. years and years ago (and it may have changed now), all the hospitals were government owned, lab personnel got a day off each week with pay to attend IMLT classes, and the classes were free. To work in a hospital laboratory you had to be "State Registered". In the USA hospitals are owned by all sorts of organizations. You don't get "day release" to attend classes. If you do attend classes, money has to come from somewhere to pay for it. Further you may live miles/hours from the nearest college where you can attend classes.Since the different organisations may be "for profit" or even "Non-profit", they want to save money by not paying too higher salaries. Most often they pay what other hospitals in the area pay for the same type work. The 50 states have different requirements for working in their hospital laboratories. Hospitals have coped with these differences by making histology laboratories a separate section of the laboratory, with technicians/technologists trained just to work in that lab. I think a histotechnologist from the U.K. could easily find a position in the USA, but they might feel underappreciated and underpaid! (maybe even under challanged!) Well, that's my two bits worth! Mike Titford USA Pathology Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Start your day with Yahoo! - make it your home page _______________________________________________ Histonet mailing list Histonet@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 Janet.Bonner <@t> FLHOSP.ORG Mon Aug 8 06:47:22 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] RE:deterioration of paraplast wax blocks Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB435F@fh2k093.fhmis.net> We had a rat problem do to the storage area.... -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 8/6/2005 7:48 AM Subject: [Histonet] RE:deterioration of paraplast wax blocks Hi All Some of our stored blocks are detoriorating due to a ? fungus which is eating them. We have asked the micro people to see if they could identify it but they have been unable to make it grow.Has anyone had a similar problem and if so how did you rectify it. Where you able to classify the organism. ian clarke Cellular Pathology Department Craigavon Area Hospital _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MLafrini <@t> csmlab.com Mon Aug 8 07:25:50 2005 From: MLafrini <@t> csmlab.com (Michael LaFriniere) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] RE:deterioration of paraplast wax blocks Message-ID: Ian, This is sad to say but you may want to check a pest control to check out your storage area, due to past experience with an off site storage area, I have found that mice and rats will eat paraffin blocks with the tissue embedded.... Good luck investigating! Michael Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax MLafrini@csmlab.com >>> "Ian Clarke" 08/06/05 7:48 AM >>> Hi All Some of our stored blocks are detoriorating due to a ? fungus which is eating them. We have asked the micro people to see if they could identify it but they have been unable to make it grow.Has anyone had a similar problem and if so how did you rectify it. Where you able to classify the organism. ian clarke Cellular Pathology Department Craigavon Area Hospital _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Mon Aug 8 07:40:34 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Sep 16 15:25:25 2005 Subject: [Histonet] RE:deterioration of paraplast wax blocks Message-ID: Janet I believe that the problem with fungi may be to leaving the surface of cut blocks unsealed. Here in Houston it is very humid and we have lot of molds, however I do not see any problem with molds in sealed blocks. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: Monday, August 08, 2005 6:47 AM To: 'Ian Clarke '; 'histonet-bounces@lists.utsouthwestern.edu '; 'histonet@lists.utsouthwestern.edu ' Subject: RE: [Histonet] RE:deterioration of paraplast wax blocks We had a rat problem do to the storage area.... -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 8/6/2005 7:48 AM Subject: [Histonet] RE:deterioration of paraplast wax blocks Hi All Some of our stored blocks are detoriorating due to a ? fungus which is eating them. We have asked the micro people to see if they could identify it but they have been unable to make it grow.Has anyone had a similar problem and if so how did you rectify it. Where you able to classify the organism. ian clarke Cellular Pathology Department Craigavon Area Hospital _______________________________________________ Histonet mailing list Histonet@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 ROrr <@t> enh.org Mon Aug 8 07:51:06 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] RE: Histonet Digest, Vol 21, Issue 8 Message-ID: Hello Everyone, Does anyone out there run Pituitary panels on a fairly regular basis? ACTH, LH, GH, FSH, PRL,and TSH... Please respond privately if you wouldn't mind having a chat with me about your experiences with them...specifically working with a single control covering all of them...I am relatively new to day-to-day clinical operations and I may be over-thinking this one... Thanks again for your help. Becky Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Sunday, August 07, 2005 11:43 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 21, Issue 8 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. tween detergens (Gudrun Lang) 2. stain (MARY JOHNSON) 3. RE: "Brit" Hitotech/Firestorm? (Charles.Embrey) 4. fixation via perfusion (Wisam Barkho) 5. Re: troubles with CD4 (Amos Brooks) 6. help: Heidanhein's azan trichrome (Sasha Manhattan) 7. RE: "Brit" Hitotech/Firestorm? (Rogerson Kemlo (ELHT) Pathology) ---------------------------------------------------------------------- Message: 1 Date: Sat, 6 Aug 2005 19:56:35 +0200 From: "Gudrun Lang" Subject: [Histonet] tween detergens To: "Histonetliste \(Histonetliste\)" Message-ID: Content-Type: text/plain; charset="us-ascii" Hi all, We are going to run FISH on our ventana benchmark. In the last step we have to wash the slides to remove the liquid cover slip. Until now we use a green dish-washing product for the ihc, but that doesn't fit with the FISH because of autofluorescence. Can anyone give me a hint, if Tween 20 in ?% solution in water is okay? Or any other well working detergens? Our Ventana-Lady is also looking for a colourless dish-washer, but with no results yet. Thank you in advance Gudrun Lang Linz, Austria ------------------------------ Message: 2 Date: Sat, 6 Aug 2005 11:16:28 -0700 (PDT) From: MARY JOHNSON Subject: [Histonet] stain To: histonet@lists.utsouthwestern.edu Message-ID: <20050806181628.33904.qmail@web81610.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Herbert, have you tried the Movat Pentachrome stain, if you need the procedure sent me your email and I will sent you the procedure. Mary ------------------------------ Message: 3 Date: Sat, 6 Aug 2005 13:55:20 -0500 From: "Charles.Embrey" Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? To: "Terry Murphy" , , Message-ID: Content-Type: text/plain; charset="us-ascii" Hi Terry, Please know that some Pathologists' Assistants belong to this list as well and I really don't like someone not in my shoes telling me that I am overpaid. I spent 20 years in the Air Force and busted my @#%@# as a Histotech and Histology Supervisor for less than underpaid civilian histotechs make. But I really wasn't in the military for the money. I finished my education and joined the AAPA, passing my Fellowship exam after retiring from the Air Force. I am here at work on a Saturday, after already putting in a 40 hour week so that I can finish up some paperwork I couldn't complete during the week because of the 500+ cases I grossed and the two cancer conferences I needed to attend. I do agree that histotechs are behind on the pay scale but don't let your "sour grapes" cause you to spout off about a profession you don't work in. Education doesn't always equate to salary. I personally know some histotechs with only high school education that can work circles around BS educated registered HTL's. They have a fully working knowledge of histology theory, do a great job and have my full respect. To me you sound like someone that "toots his own horn" a lot and probably carries mini copies of your diplomas in you wallet. I am sorry that you met an "arrogant PA" but we are not all like that. I am normally a pretty easy going, laid back southern boy except when someone spouts of generalizations that are untrue out of self pity over his own situation. Charles Embrey, HT, PA(ASCP) Histology Manager Carle Clinic Illinois -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Terry Murphy Sent: Saturday, August 06, 2005 7:34 AM To: hawkmoon15@cox.net; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? HI everyone I feel the urge to join this discussion. I got into the field of histology 15 years ago and enjoyed the work and the concepts. I wanted to move up in the field ao I earned my HTL (ASCP) instead of the HT(ASCP) since I have a Bachelor's degree in Biology. That didn't get me any more respect or money. Still trying to improve my situation I earn a Master's degree in Health Administration and I once again my employment has not improved. As a matter of fact my employment situation has gotten worse since I earned a Master's degree. I had acheived a position as a supervisor only to be "run out of the lab" by an arrogant PA and, I can only assume, seasoned histotechs that fear change. Any way now I have been working as a traveling histotech. The pay is better than if I had a permanent job. The facility I am at now recently hired a person to be trained as a tech. This new employee has a GED not a high school dipolma and he has no background in in science or medicine. He is doing good given his background but I find it kind of a insult to educated techs like myself that that someone without education or experience has been hired to do the same job that us techs have studied extensively. Now I am done traveling and am trying to get a histotech job close to my home town so I can live in my house and more importantly live with my wife. I have applied for tech positions and either haven't heard from the facility except for a rejection letter. When I can get a hold of a hireing supervisor I have been told that "this is a entry level position and it doesn't pay well", "you are overqualified", and, this really surprised me, "your skill set will not go well in our laboratory". Someone on this board mentioned about being "black listed" I feel that I have now entered that list. Most likely I have more education than the people that I would be reporting to and they probably feel threatened by me. Someone suggested that I "dumb down" my resume and exclude some education and my publication but I do not like the idea of excluding accomplishments that I am proud of. Lastly, as for wages, histotechs are grossly underpaid while Pathologist Assistants are grossly overpaid!!!! Thanks for letting me vent. Sincerely, Another fustrated histotech. >From: "Sarah Jones" >To: >Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? >Date: Fri, 5 Aug 2005 16:12:29 -0700 > >I have to remind you here, Diana, that you are speaking for yourself only >with your statement about "in NA we tend to stay put". That may be for >some >people. However, I have found throughout my ~40 year career in >Histotechnology (especially in the last half of it!!) the only way to get a >decent raise is to pack up and move on. Unfortunately, here in NA our >annual raises do not even keep up with the cost of living. In fact, it is >down-right terrible!! I have my HTL and got my CM recently, I am proud to >say. However, I haven't received any additional compensation for either, >and I doubt I ever will. (HTL for 24 yrs.) > >Which brings up something else: The ASCP used to be the American Society >of >Clinical Pathologists. It is now the American Society of Clinical >Pathology. What changed? I'm not sure. Does anyone know, except the >'insiders', that is? I do have first-hand knowledge of what used to be. >The Pathologists ran it, and they had a lot to do with where Histotechs >sat -- low on the rung of the Pathology Laboratory ladder for YEARS. The >Registry was in my home town in Muncie, Indiana, and my Mother was the >secretary to the Registrar for many years. (This was all before it was >centralized in Chicago. Prior to the relocation to Chicago, the Registry >was in the town where the Chairman of the Board was located. L. >Montgomery, >M.D., was the Chairman of the Board for ~30 years that I know of, and thus >was the Registry located in Muncie.) There are other things I could tell >you; but I was sworn to secrecy years ago, and I will honor that pledge. >Suffice it to say, the Pathologists wanted to keep the salaries of the >Histotechs down, and did everything in their power to do so. 'Nuff said! > >When I started into the field of Histotechnology at the University of >Chicago back in 1965, the Techies associated with the ASCP were trying to >change the requirements for Histotechs, but to no avail. Not until January >of 2005 did the change they were attempting to accomplish way back then >prevail. I didn't really understand what was happening then, but today I >do. There are MANY places here in the USA (where there is a severe >shortage >of Histotechs) where they will still hire techs that are not licensed. In >fact, there are places where a licensed Histologic Technician OR a licensed >Histotechnologist are black-listed by the other techs--most likely in fear >that their own incompetence might be discovered! > >The cost of living in certain areas of the country are no longer taken into >consideration in the way they were in the past when it comes to hiring >techs. That also saddens me. How is it that they think I can live in the >most expensive county in California that pays the lowest wages of any >county >in California?? That is craziness in my book!!!! Yet they wonder why they >cannot find good Histotechs here in this county! GO FIGURE!! > >Anyway, I could go on and on, but I won't. Hope this sheds some light on >some things for some folks. Please address any personal questions to: >hawkmoon15@cox.net > >Sarah > > >----- Original Message ----- >From: "Diana McCaig" >To: >Sent: Friday, August 05, 2005 10:41 AM >Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > What are you all using to compare the wage scales. Biomedical >scientists >do > > not get paid as well as we do in North America if you factor in their >cost > > of living. I understand there is a recent trend to increase the wages >in > > order to keep them on board as there is a lot of migration to various >labs > > throughout a career where in NA we tend to stay put. > > Diana McCaig, MLT > > > > > > -----Original Message----- > > From: Larry Woody [SMTP:slappycraw@yahoo.com] > > Sent: Friday, August 05, 2005 1:35 PM > > To: mtitford@aol.com; histonet@lists.utsouthwestern.edu > > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > > > You may want to try some of the big biotech companies where there is > > a need for people with more skills in the lab and the compensation can >be > > better than a medical lab. > > > > mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion > > about histotechnologist training. > > > > In regards to hospital histology labs in the USA: > > > > I don't want to start a firestorm of criticism here, but I think > > there is a real danger that in Great Britain those technologists >performing > > in the histology laboratories may be over qualified.A few months ago >here >on > > the Histonet, someone in the U.K. was bemoaning the lack of histology > > technologists in the U.K., and how short staffed they were. Was that >because > > the bar had been set too high? Was the level of education required too >much > > for the compensation received? Has a "closed shop" (to use a union >term!) >in > > effect, been created? > > Here in the United States there are hundreds of community type > > hospitals and smaller medical centers that have histology laboratories. >In > > those laboratories work histotechnologists doing a fine job with a lot >less > > education than a fully qualified histotechnologist in the U.K.. At the >other > > end of the spectrum are histology laboratories here doing more advanced > > procedures where degrees and extra training are required. > > However, histotechnology training in the U.K. and USA are inherently > > different. When I lived in the U.K. years and years ago (and it may have > > changed now), all the hospitals were government owned, lab personnel got >a > > day off each week with pay to attend IMLT classes, and the classes were > > free. To work in a hospital laboratory you had to be "State Registered". > > In the USA hospitals are owned by all sorts of organizations. You > > don't get "day release" to attend classes. If you do attend classes, >money > > has to come from somewhere to pay for it. Further you may live >miles/hours > > from the nearest college where you can attend classes.Since the >different > > organisations may be "for profit" or even "Non-profit", they want to >save > > money by not paying too higher salaries. Most often they pay what other > > hospitals in the area pay for the same type work. > > The 50 states have different requirements for working in their > > hospital laboratories. > > Hospitals have coped with these differences by making histology > > laboratories a separate section of the laboratory, with > > technicians/technologists trained just to work in that lab. > > I think a histotechnologist from the U.K. could easily find a > > position in the USA, but they might feel underappreciated and underpaid! > > (maybe even under challanged!) > > Well, that's my two bits worth! > > > > Mike Titford > > USA Pathology > > Mobile AL USA > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > --------------------------------- > > Start your day with Yahoo! - make it your home page > > _______________________________________________ > > Histonet mailing list > > Histonet@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 _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Sat, 6 Aug 2005 16:44:42 -0700 From: "Wisam Barkho" Subject: [Histonet] fixation via perfusion To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hello, We are doing stereotaxic surgeries on rats in my lab. After 2 weeks, we fixate the brain via perfusion using 4% PFA (after vascular rinse) and then immerse the brains in 4% PFA overnight. My question is where can I find information on pump flow rate and PFA volume to use during fixation? Thanks. Wisam Barkho daufoi@msn.com ------------------------------ Message: 5 Date: Sat, 06 Aug 2005 22:34:31 -0400 From: Amos Brooks Subject: [Histonet] Re: troubles with CD4 To: histonet@lists.utsouthwestern.edu Message-ID: <42F57337.3000605@earthlink.net> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Tom, What is the pH of the retrieval solution that you are using? In our experience CD4 tends to work best with a high pH retrieval solution (like Dako's high pH retrieval solution or Biocare Medical's Borg Decloaker). We use Sigma's Impress for detection of this because of it being a polymer which does not use avidin/ biotin it avoids the endogenous biotin background that high pH retrieval tends to increase. Just some suggestions to try, Amos Brooks Message: 15 Date: Fri, 05 Aug 2005 10:47:22 -0500 From: "Thomas Pier" Subject: [Histonet] troubles with CD4 To: Message-ID: Content-Type: text/plain; charset=US-ASCII Hello all, I was wondering if anyone could help me with some problems that I'm having with staining for CD4. My control tissue for the work up is formalin fixted, paraffin embeded tonsil. I'm using Labvision's CD4 Ab-2 clone 1F6. I have diluted it anywhere from 1:10-1:50. I'm using 1% goat serum in TBS as my diluent. I've tried EDTA HIER as recomended on the data sheet. I've also tried protease XXV w/ or w/o the EDTA. I shouldn't be running into the peroxidase problem since I'm using an alkaline phosphatase detection system (Biocare's Vulcan Fast Red). I have also tried using or not using a 10% goat serum protein block. I have gotten some staining, but nowhere near as much as I should in a tonsil. Any help that you could give me would be greatly appreciated. Tom Pier ------------------------------ Message: 6 Date: Sun, 7 Aug 2005 00:39:35 -0700 (PDT) From: Sasha Manhattan Subject: [Histonet] help: Heidanhein's azan trichrome To: histonet@lists.utsouthwestern.edu Message-ID: <20050807073935.7707.qmail@web31201.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello colleagues, I am a student at the University of Hong Kong. I am trying Heidanhein's azan staining to quantitate the extent of fibrosis in mouse liver. When I tried it for the first time, there was a blue colour in the tissues showing collagen deposition, which is what I expected, but the protocol I used does not include aniline. I only used 95% alcohol to extract azaocarmine G from collagen because our lab do not have aniline at the moment. but since i got the blue colour, i think it was ok. but now that I repeat it with the same tissue, i dont see any blue colour. I thought maybe the distilled water rinse washed out the water-soluble aniline blue, but i repeated without washing in distilled water (just put in 95% alcohol and then absolute alcohol and then xylene clear), but there's still no blue colour. Could you tell me what could be wrong? This is my protocol: 1.deparaffination, hydration through alcohols. 2. Put slides into azocarmine G solution at 50 degrees celsius for 1 hour. 3. Rinse quickly with dH2O. 4. Place into 95% ethanol. 5. Place into 1% acetic acid in 95% ethanol. 6. Rinse briefly in dH2O. 7. Place into 5% phosphotungstic acid for 2 hrs. 8. Place into aniline blue-orange G solution for 2 hrs. 9. Rinse very quickly with dH2O, dehydrate with 95% and 100% ethanol(just dip), 3 changes of xylene each of 3 minutes. mount. I would be very grateful if you could just tell me what might have gone wrong. There is no one for me to ask in the lab because no one does histology here, and I am really nervous and frustrated. Thank you very much for your time and attention. Yours sincerely, Mimi Kwun-nok Man --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour ------------------------------ Message: 7 Date: Sat, 6 Aug 2005 09:01:17 +0100 From: "Rogerson Kemlo \(ELHT\) Pathology" Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? To: , Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4D2@elht-exch1.xelht.nhs.uk> Content-Type: text/plain; charset="us-ascii" Thank you for that; what you say is very interesting and I agree with a lot of it. Interestingly, whether it's because it's Holiday time, no-one from the UK has answered my implied question. Are we overqualified in the UK to be 'HistoTechs'; even more contentious, what is the role of the Medical Laboratory assistant (Higher or even Associate Practitioner)? Are they US Histotechs in disguise? Do we (Biomedical Scientists) therefore, not exist in the US, and why not? Kemlo Rogerson MSc MIBiol CBiol CSci DMS FIBMS -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of mtitford@aol.com Sent: 05 August 2005 17:59 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] "Brit" Hitotech/Firestorm? Rogerson Kemlo carries on the discussion about histotechnologist training. In regards to hospital histology labs in the USA: I don't want to start a firestorm of criticism here, but I think there is a real danger that in Great Britain those technologists performing in the histology laboratories may be over qualified.A few months ago here on the Histonet, someone in the U.K. was bemoaning the lack of histology technologists in the U.K., and how short staffed they were. Was that because the bar had been set too high? Was the level of education required too much for the compensation received? Has a "closed shop" (to use a union term!) in effect, been created? Here in the United States there are hundreds of community type hospitals and smaller medical centers that have histology laboratories. In those laboratories work histotechnologists doing a fine job with a lot less education than a fully qualified histotechnologist in the U.K.. At the other end of the spectrum are histology laboratories here doing more advanced procedures where degrees and extra training are required. However, histotechnology training in the U.K. and USA are inherently different. When I lived in the U.K. years and years ago (and it may have changed now), all the hospitals were government owned, lab personnel got a day off each week with pay to attend IMLT classes, and the classes were free. To work in a hospital laboratory you had to be "State Registered". In the USA hospitals are owned by all sorts of organizations. You don't get "day release" to attend classes. If you do attend classes, money has to come from somewhere to pay for it. Further you may live miles/hours from the nearest college where you can attend classes.Since the different organisations may be "for profit" or even "Non-profit", they want to save money by not paying too higher salaries. Most often they pay what other hospitals in the area pay for the same type work. The 50 states have different requirements for working in their hospital laboratories. Hospitals have coped with these differences by making histology laboratories a separate section of the laboratory, with technicians/technologists trained just to work in that lab. I think a histotechnologist from the U.K. could easily find a position in the USA, but they might feel underappreciated and underpaid! (maybe even under challanged!) Well, that's my two bits worth! Mike Titford USA Pathology Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 21, Issue 8 *************************************** From Barry.R.Rittman <@t> uth.tmc.edu Mon Aug 8 07:53:06 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: The major problem with the situation in the USA re histotech that I see ... There is a shortage of histotechs and instead of improving the salary and career prospects many health organizations are hiring people who are not trained and will work for much less, keeping the overall salary base low. Cannot blame the people who are taking these jobs as they have to have some work. This situation will not change unless we have some attitude changes in management and in the training that is available. I personally would prefer the NSH to have its own testing and its own testing centers, for all histotechs at a certain level to have to be certified and for all individuals below this level to be in formal training programs. This requires some sort of standardization for training and testing across the country. While I hate the thought of the federal government being in charge of anything, I think that this is the only way in which respect for the profession and salaries will increase. Finally we have a big public relations gap. Very few members of the general public have any idea what histotechs do. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Terry Murphy Sent: Saturday, August 06, 2005 7:34 AM To: hawkmoon15@cox.net; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? HI everyone I feel the urge to join this discussion. I got into the field of histology 15 years ago and enjoyed the work and the concepts. I wanted to move up in the field ao I earned my HTL (ASCP) instead of the HT(ASCP) since I have a Bachelor's degree in Biology. That didn't get me any more respect or money. Still trying to improve my situation I earn a Master's degree in Health Administration and I once again my employment has not improved. As a matter of fact my employment situation has gotten worse since I earned a Master's degree. I had acheived a position as a supervisor only to be "run out of the lab" by an arrogant PA and, I can only assume, seasoned histotechs that fear change. Any way now I have been working as a traveling histotech. The pay is better than if I had a permanent job. The facility I am at now recently hired a person to be trained as a tech. This new employee has a GED not a high school dipolma and he has no background in in science or medicine. He is doing good given his background but I find it kind of a insult to educated techs like myself that that someone without education or experience has been hired to do the same job that us techs have studied extensively. Now I am done traveling and am trying to get a histotech job close to my home town so I can live in my house and more importantly live with my wife. I have applied for tech positions and either haven't heard from the facility except for a rejection letter. When I can get a hold of a hireing supervisor I have been told that "this is a entry level position and it doesn't pay well", "you are overqualified", and, this really surprised me, "your skill set will not go well in our laboratory". Someone on this board mentioned about being "black listed" I feel that I have now entered that list. Most likely I have more education than the people that I would be reporting to and they probably feel threatened by me. Someone suggested that I "dumb down" my resume and exclude some education and my publication but I do not like the idea of excluding accomplishments that I am proud of. Lastly, as for wages, histotechs are grossly underpaid while Pathologist Assistants are grossly overpaid!!!! Thanks for letting me vent. Sincerely, Another fustrated histotech. >From: "Sarah Jones" >To: >Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? >Date: Fri, 5 Aug 2005 16:12:29 -0700 > >I have to remind you here, Diana, that you are speaking for yourself only >with your statement about "in NA we tend to stay put". That may be for >some >people. However, I have found throughout my ~40 year career in >Histotechnology (especially in the last half of it!!) the only way to get a >decent raise is to pack up and move on. Unfortunately, here in NA our >annual raises do not even keep up with the cost of living. In fact, it is >down-right terrible!! I have my HTL and got my CM recently, I am proud to >say. However, I haven't received any additional compensation for either, >and I doubt I ever will. (HTL for 24 yrs.) > >Which brings up something else: The ASCP used to be the American Society >of >Clinical Pathologists. It is now the American Society of Clinical >Pathology. What changed? I'm not sure. Does anyone know, except the >'insiders', that is? I do have first-hand knowledge of what used to be. >The Pathologists ran it, and they had a lot to do with where Histotechs >sat -- low on the rung of the Pathology Laboratory ladder for YEARS. The >Registry was in my home town in Muncie, Indiana, and my Mother was the >secretary to the Registrar for many years. (This was all before it was >centralized in Chicago. Prior to the relocation to Chicago, the Registry >was in the town where the Chairman of the Board was located. L. >Montgomery, >M.D., was the Chairman of the Board for ~30 years that I know of, and thus >was the Registry located in Muncie.) There are other things I could tell >you; but I was sworn to secrecy years ago, and I will honor that pledge. >Suffice it to say, the Pathologists wanted to keep the salaries of the >Histotechs down, and did everything in their power to do so. 'Nuff said! > >When I started into the field of Histotechnology at the University of >Chicago back in 1965, the Techies associated with the ASCP were trying to >change the requirements for Histotechs, but to no avail. Not until January >of 2005 did the change they were attempting to accomplish way back then >prevail. I didn't really understand what was happening then, but today I >do. There are MANY places here in the USA (where there is a severe >shortage >of Histotechs) where they will still hire techs that are not licensed. In >fact, there are places where a licensed Histologic Technician OR a licensed >Histotechnologist are black-listed by the other techs--most likely in fear >that their own incompetence might be discovered! > >The cost of living in certain areas of the country are no longer taken into >consideration in the way they were in the past when it comes to hiring >techs. That also saddens me. How is it that they think I can live in the >most expensive county in California that pays the lowest wages of any >county >in California?? That is craziness in my book!!!! Yet they wonder why they >cannot find good Histotechs here in this county! GO FIGURE!! > >Anyway, I could go on and on, but I won't. Hope this sheds some light on >some things for some folks. Please address any personal questions to: >hawkmoon15@cox.net > >Sarah > > >----- Original Message ----- >From: "Diana McCaig" >To: >Sent: Friday, August 05, 2005 10:41 AM >Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > What are you all using to compare the wage scales. Biomedical >scientists >do > > not get paid as well as we do in North America if you factor in their >cost > > of living. I understand there is a recent trend to increase the wages >in > > order to keep them on board as there is a lot of migration to various >labs > > throughout a career where in NA we tend to stay put. > > Diana McCaig, MLT > > > > > > -----Original Message----- > > From: Larry Woody [SMTP:slappycraw@yahoo.com] > > Sent: Friday, August 05, 2005 1:35 PM > > To: mtitford@aol.com; histonet@lists.utsouthwestern.edu > > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > > > You may want to try some of the big biotech companies where there is > > a need for people with more skills in the lab and the compensation can >be > > better than a medical lab. > > > > mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion > > about histotechnologist training. > > > > In regards to hospital histology labs in the USA: > > > > I don't want to start a firestorm of criticism here, but I think > > there is a real danger that in Great Britain those technologists >performing > > in the histology laboratories may be over qualified.A few months ago >here >on > > the Histonet, someone in the U.K. was bemoaning the lack of histology > > technologists in the U.K., and how short staffed they were. Was that >because > > the bar had been set too high? Was the level of education required too >much > > for the compensation received? Has a "closed shop" (to use a union >term!) >in > > effect, been created? > > Here in the United States there are hundreds of community type > > hospitals and smaller medical centers that have histology laboratories. >In > > those laboratories work histotechnologists doing a fine job with a lot >less > > education than a fully qualified histotechnologist in the U.K.. At the >other > > end of the spectrum are histology laboratories here doing more advanced > > procedures where degrees and extra training are required. > > However, histotechnology training in the U.K. and USA are inherently > > different. When I lived in the U.K. years and years ago (and it may have > > changed now), all the hospitals were government owned, lab personnel got >a > > day off each week with pay to attend IMLT classes, and the classes were > > free. To work in a hospital laboratory you had to be "State Registered". > > In the USA hospitals are owned by all sorts of organizations. You > > don't get "day release" to attend classes. If you do attend classes, >money > > has to come from somewhere to pay for it. Further you may live >miles/hours > > from the nearest college where you can attend classes.Since the >different > > organisations may be "for profit" or even "Non-profit", they want to >save > > money by not paying too higher salaries. Most often they pay what other > > hospitals in the area pay for the same type work. > > The 50 states have different requirements for working in their > > hospital laboratories. > > Hospitals have coped with these differences by making histology > > laboratories a separate section of the laboratory, with > > technicians/technologists trained just to work in that lab. > > I think a histotechnologist from the U.K. could easily find a > > position in the USA, but they might feel underappreciated and underpaid! > > (maybe even under challanged!) > > Well, that's my two bits worth! > > > > Mike Titford > > USA Pathology > > Mobile AL USA > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > --------------------------------- > > Start your day with Yahoo! - make it your home page > > _______________________________________________ > > Histonet mailing list > > Histonet@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 _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rolmsche <@t> unlnotes.unl.edu Mon Aug 8 07:58:32 2005 From: rolmsche <@t> unlnotes.unl.edu (Robin Olmscheid) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] address Message-ID: <4.1.20050808075658.00b13200@unlnotes01.unl.edu> Does anyone have an e-mail address or phone number for Lamar Jones? Thanks From ROrr <@t> enh.org Mon Aug 8 08:06:31 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Tween on Ventana Fish Message-ID: Hi Gudrun, We are currently running Her2 Fish on our Ventana with 100% correlation with other vendor's chemistries. This is what works very well in my lab: 1)Under subdued lighting, I remove the slides and place them in 2 changes Reaction buffer. 2)Soak in the second change of reaction buffer for 5 minutes. I believe there is some kind of surfactant in the Reaction buffer that does the trick. 3)Rinse for a few minutes in running DI water if you have it...I would only use tap water if you "trust" it! 4)Soak slides in the 2Xssc for 5 minutes and coverslip out of the SSC with the Propidium Iodine. As our control sections are on the same slide as the patient, I use 10 microliters for the control area and 10 microliters on the patient area and coverslip with 25x50 coverslip. The slides are usually read the same day and are stored in the freezer until the Docs come to read them. We also cut an Extra H/E for them to read along with the FISH. Are you set up with filters for your microscope? I can help with that, too, if needed. Good luck Becky Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 From Kemlo.Rogerson <@t> elht.nhs.uk Mon Aug 8 08:13:10 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698E79@elht-exch1.xelht.nhs.uk> With respect, rather than keep complaining why don't you mobilise and form an Organisation that can best champion your causes by presenting a united front? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R Sent: 08 August 2005 13:53 To: histonet Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? The major problem with the situation in the USA re histotech that I see ... There is a shortage of histotechs and instead of improving the salary and career prospects many health organizations are hiring people who are not trained and will work for much less, keeping the overall salary base low. Cannot blame the people who are taking these jobs as they have to have some work. This situation will not change unless we have some attitude changes in management and in the training that is available. I personally would prefer the NSH to have its own testing and its own testing centers, for all histotechs at a certain level to have to be certified and for all individuals below this level to be in formal training programs. This requires some sort of standardization for training and testing across the country. While I hate the thought of the federal government being in charge of anything, I think that this is the only way in which respect for the profession and salaries will increase. Finally we have a big public relations gap. Very few members of the general public have any idea what histotechs do. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Terry Murphy Sent: Saturday, August 06, 2005 7:34 AM To: hawkmoon15@cox.net; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? HI everyone I feel the urge to join this discussion. I got into the field of histology 15 years ago and enjoyed the work and the concepts. I wanted to move up in the field ao I earned my HTL (ASCP) instead of the HT(ASCP) since I have a Bachelor's degree in Biology. That didn't get me any more respect or money. Still trying to improve my situation I earn a Master's degree in Health Administration and I once again my employment has not improved. As a matter of fact my employment situation has gotten worse since I earned a Master's degree. I had acheived a position as a supervisor only to be "run out of the lab" by an arrogant PA and, I can only assume, seasoned histotechs that fear change. Any way now I have been working as a traveling histotech. The pay is better than if I had a permanent job. The facility I am at now recently hired a person to be trained as a tech. This new employee has a GED not a high school dipolma and he has no background in in science or medicine. He is doing good given his background but I find it kind of a insult to educated techs like myself that that someone without education or experience has been hired to do the same job that us techs have studied extensively. Now I am done traveling and am trying to get a histotech job close to my home town so I can live in my house and more importantly live with my wife. I have applied for tech positions and either haven't heard from the facility except for a rejection letter. When I can get a hold of a hireing supervisor I have been told that "this is a entry level position and it doesn't pay well", "you are overqualified", and, this really surprised me, "your skill set will not go well in our laboratory". Someone on this board mentioned about being "black listed" I feel that I have now entered that list. Most likely I have more education than the people that I would be reporting to and they probably feel threatened by me. Someone suggested that I "dumb down" my resume and exclude some education and my publication but I do not like the idea of excluding accomplishments that I am proud of. Lastly, as for wages, histotechs are grossly underpaid while Pathologist Assistants are grossly overpaid!!!! Thanks for letting me vent. Sincerely, Another fustrated histotech. >From: "Sarah Jones" >To: >Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? >Date: Fri, 5 Aug 2005 16:12:29 -0700 > >I have to remind you here, Diana, that you are speaking for yourself only >with your statement about "in NA we tend to stay put". That may be for >some >people. However, I have found throughout my ~40 year career in >Histotechnology (especially in the last half of it!!) the only way to get a >decent raise is to pack up and move on. Unfortunately, here in NA our >annual raises do not even keep up with the cost of living. In fact, it is >down-right terrible!! I have my HTL and got my CM recently, I am proud to >say. However, I haven't received any additional compensation for either, >and I doubt I ever will. (HTL for 24 yrs.) > >Which brings up something else: The ASCP used to be the American Society >of >Clinical Pathologists. It is now the American Society of Clinical >Pathology. What changed? I'm not sure. Does anyone know, except the >'insiders', that is? I do have first-hand knowledge of what used to be. >The Pathologists ran it, and they had a lot to do with where Histotechs >sat -- low on the rung of the Pathology Laboratory ladder for YEARS. The >Registry was in my home town in Muncie, Indiana, and my Mother was the >secretary to the Registrar for many years. (This was all before it was >centralized in Chicago. Prior to the relocation to Chicago, the Registry >was in the town where the Chairman of the Board was located. L. >Montgomery, >M.D., was the Chairman of the Board for ~30 years that I know of, and thus >was the Registry located in Muncie.) There are other things I could tell >you; but I was sworn to secrecy years ago, and I will honor that pledge. >Suffice it to say, the Pathologists wanted to keep the salaries of the >Histotechs down, and did everything in their power to do so. 'Nuff said! > >When I started into the field of Histotechnology at the University of >Chicago back in 1965, the Techies associated with the ASCP were trying to >change the requirements for Histotechs, but to no avail. Not until January >of 2005 did the change they were attempting to accomplish way back then >prevail. I didn't really understand what was happening then, but today I >do. There are MANY places here in the USA (where there is a severe >shortage >of Histotechs) where they will still hire techs that are not licensed. In >fact, there are places where a licensed Histologic Technician OR a licensed >Histotechnologist are black-listed by the other techs--most likely in fear >that their own incompetence might be discovered! > >The cost of living in certain areas of the country are no longer taken into >consideration in the way they were in the past when it comes to hiring >techs. That also saddens me. How is it that they think I can live in the >most expensive county in California that pays the lowest wages of any >county >in California?? That is craziness in my book!!!! Yet they wonder why they >cannot find good Histotechs here in this county! GO FIGURE!! > >Anyway, I could go on and on, but I won't. Hope this sheds some light on >some things for some folks. Please address any personal questions to: >hawkmoon15@cox.net > >Sarah > > >----- Original Message ----- >From: "Diana McCaig" >To: >Sent: Friday, August 05, 2005 10:41 AM >Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > What are you all using to compare the wage scales. Biomedical >scientists >do > > not get paid as well as we do in North America if you factor in their >cost > > of living. I understand there is a recent trend to increase the wages >in > > order to keep them on board as there is a lot of migration to various >labs > > throughout a career where in NA we tend to stay put. > > Diana McCaig, MLT > > > > > > -----Original Message----- > > From: Larry Woody [SMTP:slappycraw@yahoo.com] > > Sent: Friday, August 05, 2005 1:35 PM > > To: mtitford@aol.com; histonet@lists.utsouthwestern.edu > > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > > > You may want to try some of the big biotech companies where there is > > a need for people with more skills in the lab and the compensation can >be > > better than a medical lab. > > > > mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion > > about histotechnologist training. > > > > In regards to hospital histology labs in the USA: > > > > I don't want to start a firestorm of criticism here, but I think > > there is a real danger that in Great Britain those technologists >performing > > in the histology laboratories may be over qualified.A few months ago >here >on > > the Histonet, someone in the U.K. was bemoaning the lack of histology > > technologists in the U.K., and how short staffed they were. Was that >because > > the bar had been set too high? Was the level of education required too >much > > for the compensation received? Has a "closed shop" (to use a union >term!) >in > > effect, been created? > > Here in the United States there are hundreds of community type > > hospitals and smaller medical centers that have histology laboratories. >In > > those laboratories work histotechnologists doing a fine job with a lot >less > > education than a fully qualified histotechnologist in the U.K.. At the >other > > end of the spectrum are histology laboratories here doing more advanced > > procedures where degrees and extra training are required. > > However, histotechnology training in the U.K. and USA are inherently > > different. When I lived in the U.K. years and years ago (and it may have > > changed now), all the hospitals were government owned, lab personnel got >a > > day off each week with pay to attend IMLT classes, and the classes were > > free. To work in a hospital laboratory you had to be "State Registered". > > In the USA hospitals are owned by all sorts of organizations. You > > don't get "day release" to attend classes. If you do attend classes, >money > > has to come from somewhere to pay for it. Further you may live >miles/hours > > from the nearest college where you can attend classes.Since the >different > > organisations may be "for profit" or even "Non-profit", they want to >save > > money by not paying too higher salaries. Most often they pay what other > > hospitals in the area pay for the same type work. > > The 50 states have different requirements for working in their > > hospital laboratories. > > Hospitals have coped with these differences by making histology > > laboratories a separate section of the laboratory, with > > technicians/technologists trained just to work in that lab. > > I think a histotechnologist from the U.K. could easily find a > > position in the USA, but they might feel underappreciated and underpaid! > > (maybe even under challanged!) > > Well, that's my two bits worth! > > > > Mike Titford > > USA Pathology > > Mobile AL USA > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > --------------------------------- > > Start your day with Yahoo! - make it your home page > > _______________________________________________ > > Histonet mailing list > > Histonet@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 _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Aug 8 10:38:50 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Concentration of detergents used with Ventana Fish In-Reply-To: References: Message-ID: <6.0.0.22.1.20050808093528.01b6d0b0@gemini.msu.montana.edu> Gudrun, It seems to me that Ventana technical services can and should supply you with this information i.e. percentage of Tween 20 or Triton X 100 or is it on the MSDS (Material Safety Data Sheets) for the Reaction buffer? At 07:06 AM 8/8/2005, you wrote: >Hi Gudrun, >We are currently running Her2 Fish on our Ventana with 100% correlation >with other vendor's chemistries. >This is what works very well in my lab: >1)Under subdued lighting, I remove the slides and place them in 2 >changes Reaction buffer. >2)Soak in the second change of reaction buffer for 5 minutes. I believe >there is some kind of surfactant in the Reaction buffer that does the >trick. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From tpmorken <@t> labvision.com Mon Aug 8 10:44:50 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Tween on Ventana Fish Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328CFD4@usca0082k08.labvision.apogent.com> Becky wrote: "1)Under subdued lighting, I remove the slides ..." And I'm imagining flickering candles and a mysteriious hooded person making incantations over their latest creation... Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Orr, Rebecca Sent: Monday, August 08, 2005 6:07 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tween on Ventana Fish Hi Gudrun, We are currently running Her2 Fish on our Ventana with 100% correlation with other vendor's chemistries. This is what works very well in my lab: 1)Under subdued lighting, I remove the slides and place them in 2 changes Reaction buffer. 2)Soak in the second change of reaction buffer for 5 minutes. I believe there is some kind of surfactant in the Reaction buffer that does the trick. 3)Rinse for a few minutes in running DI water if you have it...I would only use tap water if you "trust" it! 4)Soak slides in the 2Xssc for 5 minutes and coverslip out of the SSC with the Propidium Iodine. As our control sections are on the same slide as the patient, I use 10 microliters for the control area and 10 microliters on the patient area and coverslip with 25x50 coverslip. The slides are usually read the same day and are stored in the freezer until the Docs come to read them. We also cut an Extra H/E for them to read along with the FISH. Are you set up with filters for your microscope? I can help with that, too, if needed. Good luck Becky Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Aug 8 10:48:21 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Reference on shrinkage of bone histological preparations Message-ID: <6.0.0.22.1.20050808094005.01bb2eb0@gemini.msu.montana.edu> For those interested in publication on bone shrinkage during histological preparation, Lane J, Ralis ZA. Changes in dimensions of large cancellous bone specimens during histological preparation as measured on slabs from human femoral heads. Calcif Tissue Int (1983) 35:1-4. Although an old publication, it was interesting to note how they did the study, and measurements taken after fixation decalcification water rinses processing a. dehydration in alcohols b. clearing agent embedding in paraplast after cutting 8 and 14 um sections after staining Total shrinkage after all these factors Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From jkiernan <@t> uwo.ca Mon Aug 8 10:52:07 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:26 2005 Subject: AW: [Histonet] tween detergens References: <0IKW00JY5EHMUI10@chico.mail.uwo.pri> Message-ID: <42F77FA7.6F7CF718@uwo.ca> Gudrun Lang wrote: > If you say Tween 20 is good for washing FISH-slides, > I am satisfied and say thank you very much. That is NOT what I said! I wrote" > > ... You must > > faithfully follow the original published procedure, using the > > same kind of cells or tissues, and including proper positive > > and negative controls. ... That means you must find the original publication in a library, to be sure that you have the correct instructions for the method. The instructions you were given before (with fluorescent dishwashing liquid) obviously were wrong. That is why you must read the original publication. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ From gu.lang <@t> gmx.at Mon Aug 8 11:02:00 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:26 2005 Subject: AW: [Histonet] Concentration of detergents used with Ventana Fish In-Reply-To: <6.0.0.22.1.20050808093528.01b6d0b0@gemini.msu.montana.edu> Message-ID: Thank you all for your answers. Gudrun From David.Edmondson <@t> christie-tr.nwest.nhs.uk Mon Aug 8 11:08:56 2005 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Tween 20 @ 0.01% Message-ID: This is what we use in our PBS wash/diluent for immuno Dave Christie Manchester UK From JMacDonald <@t> mtsac.edu Mon Aug 8 11:27:57 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] address In-Reply-To: <4.1.20050808075658.00b13200@unlnotes01.unl.edu> Message-ID: mjones4@lsuhsc.edu 318-675-4912 Robin Olmscheid Sent by: histonet-bounces@lists.utsouthwestern.edu 08/08/2005 05:58 AM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] address Does anyone have an e-mail address or phone number for Lamar Jones? Thanks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bmunk <@t> uwyo.edu Mon Aug 8 12:01:33 2005 From: bmunk <@t> uwyo.edu (Brandon Munk) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Cresyl Violet stain for bird brains Message-ID: I am having trouble obtaining an acceptable Nissl stain of passerine brains using cresyl violet. The birds were transcardially perfused, brains removed, postfixed and cryoprotected before sectioning at 40 um on a sledge microtome. The protocol I am using was originally used for rat brains and is as follow: ddH20 (few dips) cresyl violet (5-10 min) ddH20 (rinse excess stain) 70% etOH (few dips) 95% etOH (few dips) 95% etOH + acetic acid (few dips) ddH20 (few dips) 95% etOH (few dips) 100% etOH (few dips) repeat previous steps as necessary Xylene (one dip) coverslip I am unsure as to the concentration of cresyl violet I am using and it has sat in a fridge for >2 yrs (although I have been assured that the age should not be a problem). I have tried playing with the protocol quite a bit, varying time in cresyl violet or how many times I dehydrate and differentiate. I am getting near my wits end and still cannot get a good stain. All my sections look as if they are washed out and I often have some stain 'smeared' under the cover slip after dipping in xylene. Can anyone help me? Thanks in advance! Brandon _____ Brandon Munk Department of Zoology and Physiology University of Wyoming PO Box 3166 Laramie, WY 82071 _____ bmunk@uwyo.edu From Matthew_Frank <@t> URMC.Rochester.edu Mon Aug 8 12:19:49 2005 From: Matthew_Frank <@t> URMC.Rochester.edu (Frank, Matthew) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Cresyl Violet stain for bird brains Message-ID: Cresyl Violet Solution: 0.02g dye, 100ml Distilled water, 0.25ml glacial acetic acid. -----Original Message----- From: Brandon Munk [mailto:bmunk@uwyo.edu] Sent: Monday, August 08, 2005 1:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cresyl Violet stain for bird brains I am having trouble obtaining an acceptable Nissl stain of passerine brains using cresyl violet. The birds were transcardially perfused, brains removed, postfixed and cryoprotected before sectioning at 40 um on a sledge microtome. The protocol I am using was originally used for rat brains and is as follow: ddH20 (few dips) cresyl violet (5-10 min) ddH20 (rinse excess stain) 70% etOH (few dips) 95% etOH (few dips) 95% etOH + acetic acid (few dips) ddH20 (few dips) 95% etOH (few dips) 100% etOH (few dips) repeat previous steps as necessary Xylene (one dip) coverslip I am unsure as to the concentration of cresyl violet I am using and it has sat in a fridge for >2 yrs (although I have been assured that the age should not be a problem). I have tried playing with the protocol quite a bit, varying time in cresyl violet or how many times I dehydrate and differentiate. I am getting near my wits end and still cannot get a good stain. All my sections look as if they are washed out and I often have some stain 'smeared' under the cover slip after dipping in xylene. Can anyone help me? Thanks in advance! Brandon _____ Brandon Munk Department of Zoology and Physiology University of Wyoming PO Box 3166 Laramie, WY 82071 _____ bmunk@uwyo.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Mon Aug 8 12:22:06 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] fixation via perfusion In-Reply-To: References: Message-ID: <42F794BE.8020906@umdnj.edu> Hi Wisam: I don't have the exact answer you are looking for but I can provide some direction. This information was probably worked out in the late 1950's and/or early 1960's as perfusion fixation emerged as the way to fix for electron microscopy. I would suggest looking in Hayat's "Principles and Techniques for Electron Microscopy" or Lillie and Fullmer's "Histological Technique and Practical Histochemistry" as places to look for the original references. In my experience, most people use too small a canula and/or too low a flow rate for good fixation. The rate of flow you use should approximate the cardiac output of the animal in question. As far as volume goes, I usually use a 3 times the weight of the animal if I am perfusing the whole body, less if you clamp off the lower body and perfuse just the head. Good luck! Geoff Wisam Barkho wrote: >Hello, > > > >We are doing stereotaxic surgeries on rats in my lab. After 2 weeks, we >fixate the brain via perfusion using 4% PFA (after vascular rinse) and then >immerse the brains in 4% PFA overnight. My question is where can I find >information on pump flow rate and PFA volume to use during fixation? Thanks. > > > >Wisam Barkho > >daufoi@msn.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 mucram11 <@t> comcast.net Mon Aug 8 12:50:44 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: <080820051750.21475.42F79B72000EE71B000053E32207020653CECE030E9D0C9A03@comcast.net> I agree wholehaeartedly with Barry. We need better PR for the field as most people have not idea what a histologist is, much less does what we do. The brief acknowledgements on the crime shows and CSI saying well this is the histology reort or more often pathology report or send it to histology or pathology will not get us any recoginition. Everyone knows blood and body fluids go to a laboratory for testing few know about surgical specimens or what is done with them. The pahologist are still happy to hire anyone who says they want to learn histology and not worry about a degree because for too long we have allowed it and ASCP (pathologist) were getting off cheap. They don't want to pay a going rate for a field we have not stood and fought for in the past. Education has remained the most important thing for med tech and yet they are more automated today and less hands on than we will ever be. We need more schools and as Barry suggests even though we all hate the word standarization we will only grow and get recognition when the field is able to show we have testing and abilities we can prove. No one wants to pay the same for a person who learned on the job as for the person who has a degree. I know many histologist who were OJT and trust them with my specimens however, the times have changed and we need to change to grow starting with NSH and the members coming up with ways to let people know who we are and what we do. Sorry if this steps on some toes however my first years in histology were all OJT and I am proud of what I learned and that I had the chance learn more over the years. Pam Marcum -------------- Original message -------------- > The major problem with the situation in the USA re histotech that I see > ... > There is a shortage of histotechs and instead of improving the salary > and career prospects many health organizations are hiring people who are > not trained and will work for much less, keeping the overall salary base > low. Cannot blame the people who are taking these jobs as they have to > have some work. > This situation will not change unless we have some attitude changes in > management and in the training that is available. > I personally would prefer the NSH to have its own testing and its own > testing centers, for all histotechs at a certain level to have to be > certified and for all individuals below this level to be in formal > training programs. This requires some sort of standardization for > training and testing across the country. While I hate the thought of the > federal government being in charge of anything, I think that this is the > only way in which respect for the profession and salaries will increase. > Finally we have a big public relations gap. Very few members of the > general public have any idea what histotechs do. > Barry > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Terry > Murphy > Sent: Saturday, August 06, 2005 7:34 AM > To: hawkmoon15@cox.net; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > HI everyone > > I feel the urge to join this discussion. I got into the field of > histology > 15 years ago and enjoyed the work and the concepts. I wanted to move up > in > the field ao I earned my HTL (ASCP) instead of the HT(ASCP) since I have > a > Bachelor's degree in Biology. That didn't get me any more respect or > money. > Still trying to improve my situation I earn a Master's degree in > Health > Administration and I once again my employment has not improved. As a > matter > of fact my employment situation has gotten worse since I earned a > Master's > degree. I had acheived a position as a supervisor only to be "run out > of > the lab" by an arrogant PA and, I can only assume, seasoned histotechs > that > fear change. > > Any way now I have been working as a traveling histotech. The pay is > better > than if I had a permanent job. The facility I am at now recently hired > a > person to be trained as a tech. This new employee has a GED not a high > school dipolma and he has no background in in science or medicine. He > is > doing good given his background but I find it kind of a insult to > educated > techs like myself that that someone without education or experience has > been > hired to do the same job that us techs have studied extensively. > > Now I am done traveling and am trying to get a histotech job close to my > > home town so I can live in my house and more importantly live with my > wife. > I have applied for tech positions and either haven't heard from the > facility > except for a rejection letter. When I can get a hold of a hireing > supervisor I have been told that "this is a entry level position and it > doesn't pay well", "you are overqualified", and, this really surprised > me, > "your skill set will not go well in our laboratory". Someone on this > board > mentioned about being "black listed" I feel that I have now entered that > > list. Most likely I have more education than the people that I would be > > reporting to and they probably feel threatened by me. Someone suggested > > that I "dumb down" my resume and exclude some education and my > publication > but I do not like the idea of excluding accomplishments that I am proud > of. > > Lastly, as for wages, histotechs are grossly underpaid while Pathologist > > Assistants are grossly overpaid!!!! > > Thanks for letting me vent. Sincerely, > > Another fustrated histotech. > > >From: "Sarah Jones" > >To: > >Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > >Date: Fri, 5 Aug 2005 16:12:29 -0700 > > > >I have to remind you here, Diana, that you are speaking for yourself > only > >with your statement about "in NA we tend to stay put". That may be for > > >some > >people. However, I have found throughout my ~40 year career in > >Histotechnology (especially in the last half of it!!) the only way to > get a > >decent raise is to pack up and move on. Unfortunately, here in NA our > >annual raises do not even keep up with the cost of living. In fact, it > is > >down-right terrible!! I have my HTL and got my CM recently, I am proud > to > >say. However, I haven't received any additional compensation for > either, > >and I doubt I ever will. (HTL for 24 yrs.) > > > >Which brings up something else: The ASCP used to be the American > Society > >of > >Clinical Pathologists. It is now the American Society of Clinical > >Pathology. What changed? I'm not sure. Does anyone know, except the > >'insiders', that is? I do have first-hand knowledge of what used to > be. > >The Pathologists ran it, and they had a lot to do with where Histotechs > >sat -- low on the rung of the Pathology Laboratory ladder for YEARS. > The > >Registry was in my home town in Muncie, Indiana, and my Mother was the > >secretary to the Registrar for many years. (This was all before it was > >centralized in Chicago. Prior to the relocation to Chicago, the > Registry > >was in the town where the Chairman of the Board was located. L. > >Montgomery, > >M.D., was the Chairman of the Board for ~30 years that I know of, and > thus > >was the Registry located in Muncie.) There are other things I could > tell > >you; but I was sworn to secrecy years ago, and I will honor that > pledge. > >Suffice it to say, the Pathologists wanted to keep the salaries of the > >Histotechs down, and did everything in their power to do so. 'Nuff > said! > > > >When I started into the field of Histotechnology at the University of > >Chicago back in 1965, the Techies associated with the ASCP were trying > to > >change the requirements for Histotechs, but to no avail. Not until > January > >of 2005 did the change they were attempting to accomplish way back then > >prevail. I didn't really understand what was happening then, but > today I > >do. There are MANY places here in the USA (where there is a severe > >shortage > >of Histotechs) where they will still hire techs that are not licensed. > In > >fact, there are places where a licensed Histologic Technician OR a > licensed > >Histotechnologist are black-listed by the other techs--most likely in > fear > >that their own incompetence might be discovered! > > > >The cost of living in certain areas of the country are no longer taken > into > >consideration in the way they were in the past when it comes to hiring > >techs. That also saddens me. How is it that they think I can live in > the > >most expensive county in California that pays the lowest wages of any > >county > >in California?? That is craziness in my book!!!! Yet they wonder why > they > >cannot find good Histotechs here in this county! GO FIGURE!! > > > >Anyway, I could go on and on, but I won't. Hope this sheds some light > on > >some things for some folks. Please address any personal questions to: > >hawkmoon15@cox.net > > > >Sarah > > > > > >----- Original Message ----- > >From: "Diana McCaig" > >To: > >Sent: Friday, August 05, 2005 10:41 AM > >Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > > > > > What are you all using to compare the wage scales. Biomedical > >scientists > >do > > > not get paid as well as we do in North America if you factor in > their > >cost > > > of living. I understand there is a recent trend to increase the > wages > >in > > > order to keep them on board as there is a lot of migration to > various > >labs > > > throughout a career where in NA we tend to stay put. > > > Diana McCaig, MLT > > > > > > > > > -----Original Message----- > > > From: Larry Woody [SMTP:slappycraw@yahoo.com] > > > Sent: Friday, August 05, 2005 1:35 PM > > > To: mtitford@aol.com; histonet@lists.utsouthwestern.edu > > > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > You may want to try some of the big biotech companies where there is > > > a need for people with more skills in the lab and the compensation > can > >be > > > better than a medical lab. > > > > > > mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion > > > about histotechnologist training. > > > > > > In regards to hospital histology labs in the USA: > > > > > > I don't want to start a firestorm of criticism here, but I think > > > there is a real danger that in Great Britain those technologists > >performing > > > in the histology laboratories may be over qualified.A few months ago > > >here > >on > > > the Histonet, someone in the U.K. was bemoaning the lack of > histology > > > technologists in the U.K., and how short staffed they were. Was that > >because > > > the bar had been set too high? Was the level of education required > too > >much > > > for the compensation received? Has a "closed shop" (to use a union > >term!) > >in > > > effect, been created? > > > Here in the United States there are hundreds of community type > > > hospitals and smaller medical centers that have histology > laboratories. > >In > > > those laboratories work histotechnologists doing a fine job with a > lot > >less > > > education than a fully qualified histotechnologist in the U.K.. At > the > >other > > > end of the spectrum are histology laboratories here doing more > advanced > > > procedures where degrees and extra training are required. > > > However, histotechnology training in the U.K. and USA are inherently > > > different. When I lived in the U.K. years and years ago (and it may > have > > > changed now), all the hospitals were government owned, lab personnel > got > >a > > > day off each week with pay to attend IMLT classes, and the classes > were > > > free. To work in a hospital laboratory you had to be "State > Registered". > > > In the USA hospitals are owned by all sorts of organizations. You > > > don't get "day release" to attend classes. If you do attend classes, > > >money > > > has to come from somewhere to pay for it. Further you may live > >miles/hours > > > from the nearest college where you can attend classes.Since the > >different > > > organisations may be "for profit" or even "Non-profit", they want to > > >save > > > money by not paying too higher salaries. Most often they pay what > other > > > hospitals in the area pay for the same type work. > > > The 50 states have different requirements for working in their > > > hospital laboratories. > > > Hospitals have coped with these differences by making histology > > > laboratories a separate section of the laboratory, with > > > technicians/technologists trained just to work in that lab. > > > I think a histotechnologist from the U.K. could easily find a > > > position in the USA, but they might feel underappreciated and > underpaid! > > > (maybe even under challanged!) > > > Well, that's my two bits worth! > > > > > > Mike Titford > > > USA Pathology > > > Mobile AL USA > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > > > > --------------------------------- > > > Start your day with Yahoo! - make it your home page > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@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 > > _________________________________________________________________ > Express yourself instantly with MSN Messenger! Download today - it's > FREE! > http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PIXLEYSK <@t> UCMAIL.UC.EDU Mon Aug 8 12:56:58 2005 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Rat perfusion rates Message-ID: Dear Wisam Barkho: We have done rat perfusion and here is our procedure. We perfuse cold saline until the liver blanches (turns distinctly and evenly paler) and the fluid coming out of the heart is clear. This is about 5 mins. Then we switch to ice-cold 4% para and let it go for 30 minutes. In our hands, this is almost, but not quite 500 mls. So we perfuse at ~one liter/hr. For us, with our tubing and pump, if the flow is right, then it just drips out of the perfusion needle, it does not squirt in a stream. And after inserting the needle, you definitely do not want to see fluid dripping from the nose. That is too much pressure. A colleague here, who does perfusion even more often, does both the saline and 500 mls of para in a total of 30 mins, so they are running a bit faster. The best guide is to examine the tissues. Too great a flow will result in expansion of the blood vessels. This may be difficult to determine if you are not used to looking at your tissue without perfusion. As for amounts, we use 500 mls per adult rat. Sincerely, Sarah Pixley University of Cincinnati College of Medicine From jqb7 <@t> cdc.gov Mon Aug 8 13:04:05 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: I keep a supply of the brochures that can be purchased from NSH: "The Art and Science of Histotechnology, A Career to Consider". This brochure will assist individuals in evaluating career opportunities as a histotechnologist. One copy free. $8.00 postage/handling for quantities of 25. I bought 6 of the packs of 25. When my son was in high school I took some by and gave them to his anatomy teacher to pass around. I have taken them to career days in the school as well. If we all take a little time and a few dollars we can help ourselves by spreading the word. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 (404) 639-3590 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of mucram11@comcast.net Sent: Monday, August 08, 2005 1:51 PM To: Rittman, Barry R; histonet Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? I agree wholehaeartedly with Barry. We need better PR for the field as most people have not idea what a histologist is, much less does what we do. The brief acknowledgements on the crime shows and CSI saying well this is the histology reort or more often pathology report or send it to histology or pathology will not get us any recoginition. Everyone knows blood and body fluids go to a laboratory for testing few know about surgical specimens or what is done with them. The pahologist are still happy to hire anyone who says they want to learn histology and not worry about a degree because for too long we have allowed it and ASCP (pathologist) were getting off cheap. They don't want to pay a going rate for a field we have not stood and fought for in the past. Education has remained the most important thing for med tech and yet they are more automated today and less hands on than we will ever be. We need more schools and as Barry suggests even though we all hate the word standarization we will only grow and get recognition when the field is able to show we have testing and abilities we can prove. No one wants to pay the same for a person who learned on the job as for the person who has a degree. I know many histologist who were OJT and trust them with my specimens however, the times have changed and we need to change to grow starting with NSH and the members coming up with ways to let people know who we are and what we do. Sorry if this steps on some toes however my first years in histology were all OJT and I am proud of what I learned and that I had the chance learn more over the years. Pam Marcum -------------- Original message -------------- > The major problem with the situation in the USA re histotech that I > see ... > There is a shortage of histotechs and instead of improving the salary > and career prospects many health organizations are hiring people who > are not trained and will work for much less, keeping the overall > salary base low. Cannot blame the people who are taking these jobs as > they have to have some work. > This situation will not change unless we have some attitude changes in > management and in the training that is available. > I personally would prefer the NSH to have its own testing and its own > testing centers, for all histotechs at a certain level to have to be > certified and for all individuals below this level to be in formal > training programs. This requires some sort of standardization for > training and testing across the country. While I hate the thought of > the federal government being in charge of anything, I think that this > is the only way in which respect for the profession and salaries will increase. > Finally we have a big public relations gap. Very few members of the > general public have any idea what histotechs do. > Barry > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Terry > Murphy > Sent: Saturday, August 06, 2005 7:34 AM > To: hawkmoon15@cox.net; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > HI everyone > > I feel the urge to join this discussion. I got into the field of > histology > 15 years ago and enjoyed the work and the concepts. I wanted to move > up in the field ao I earned my HTL (ASCP) instead of the HT(ASCP) > since I have a Bachelor's degree in Biology. That didn't get me any > more respect or money. > Still trying to improve my situation I earn a Master's degree in > Health Administration and I once again my employment has not improved. > As a matter of fact my employment situation has gotten worse since I > earned a Master's degree. I had acheived a position as a supervisor > only to be "run out of the lab" by an arrogant PA and, I can only > assume, seasoned histotechs that fear change. > > Any way now I have been working as a traveling histotech. The pay is > better than if I had a permanent job. The facility I am at now > recently hired a person to be trained as a tech. This new employee has > a GED not a high school dipolma and he has no background in in science > or medicine. He is doing good given his background but I find it kind > of a insult to educated techs like myself that that someone without > education or experience has been hired to do the same job that us > techs have studied extensively. > > Now I am done traveling and am trying to get a histotech job close to > my > > home town so I can live in my house and more importantly live with my > wife. > I have applied for tech positions and either haven't heard from the > facility except for a rejection letter. When I can get a hold of a > hireing supervisor I have been told that "this is a entry level > position and it doesn't pay well", "you are overqualified", and, this > really surprised me, "your skill set will not go well in our > laboratory". Someone on this board mentioned about being "black > listed" I feel that I have now entered that > > list. Most likely I have more education than the people that I would > be > > reporting to and they probably feel threatened by me. Someone > suggested > > that I "dumb down" my resume and exclude some education and my > publication but I do not like the idea of excluding accomplishments > that I am proud of. > > Lastly, as for wages, histotechs are grossly underpaid while > Pathologist > > Assistants are grossly overpaid!!!! > > Thanks for letting me vent. Sincerely, > > Another fustrated histotech. > > >From: "Sarah Jones" > >To: > >Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > >Date: Fri, 5 Aug 2005 16:12:29 -0700 > > > >I have to remind you here, Diana, that you are speaking for yourself > only > >with your statement about "in NA we tend to stay put". That may be > >for > > >some > >people. However, I have found throughout my ~40 year career in > >Histotechnology (especially in the last half of it!!) the only way to > get a > >decent raise is to pack up and move on. Unfortunately, here in NA our > >annual raises do not even keep up with the cost of living. In fact, > >it > is > >down-right terrible!! I have my HTL and got my CM recently, I am > >proud > to > >say. However, I haven't received any additional compensation for > either, > >and I doubt I ever will. (HTL for 24 yrs.) > > > >Which brings up something else: The ASCP used to be the American > Society > >of > >Clinical Pathologists. It is now the American Society of Clinical > >Pathology. What changed? I'm not sure. Does anyone know, except the > >'insiders', that is? I do have first-hand knowledge of what used to > be. > >The Pathologists ran it, and they had a lot to do with where > >Histotechs sat -- low on the rung of the Pathology Laboratory ladder for YEARS. > The > >Registry was in my home town in Muncie, Indiana, and my Mother was > >the secretary to the Registrar for many years. (This was all before > >it was centralized in Chicago. Prior to the relocation to Chicago, > >the > Registry > >was in the town where the Chairman of the Board was located. L. > >Montgomery, > >M.D., was the Chairman of the Board for ~30 years that I know of, and > thus > >was the Registry located in Muncie.) There are other things I could > tell > >you; but I was sworn to secrecy years ago, and I will honor that > pledge. > >Suffice it to say, the Pathologists wanted to keep the salaries of > >the Histotechs down, and did everything in their power to do so. > >'Nuff > said! > > > >When I started into the field of Histotechnology at the University of > >Chicago back in 1965, the Techies associated with the ASCP were > >trying > to > >change the requirements for Histotechs, but to no avail. Not until > January > >of 2005 did the change they were attempting to accomplish way back > >then prevail. I didn't really understand what was happening then, but > today I > >do. There are MANY places here in the USA (where there is a severe > >shortage of Histotechs) where they will still hire techs that are not > >licensed. > In > >fact, there are places where a licensed Histologic Technician OR a > licensed > >Histotechnologist are black-listed by the other techs--most likely in > fear > >that their own incompetence might be discovered! > > > >The cost of living in certain areas of the country are no longer > >taken > into > >consideration in the way they were in the past when it comes to > >hiring techs. That also saddens me. How is it that they think I can > >live in > the > >most expensive county in California that pays the lowest wages of any > >county in California?? That is craziness in my book!!!! Yet they > >wonder why > they > >cannot find good Histotechs here in this county! GO FIGURE!! > > > >Anyway, I could go on and on, but I won't. Hope this sheds some light > on > >some things for some folks. Please address any personal questions to: > >hawkmoon15@cox.net > > > >Sarah > > > > > >----- Original Message ----- > >From: "Diana McCaig" > >To: > >Sent: Friday, August 05, 2005 10:41 AM > >Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > > > > > What are you all using to compare the wage scales. Biomedical > >scientists > >do > > > not get paid as well as we do in North America if you factor in > their > >cost > > > of living. I understand there is a recent trend to increase the > wages > >in > > > order to keep them on board as there is a lot of migration to > various > >labs > > > throughout a career where in NA we tend to stay put. > > > Diana McCaig, MLT > > > > > > > > > -----Original Message----- > > > From: Larry Woody [SMTP:slappycraw@yahoo.com] > > > Sent: Friday, August 05, 2005 1:35 PM > > > To: mtitford@aol.com; histonet@lists.utsouthwestern.edu > > > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > You may want to try some of the big biotech companies where there > > > is a need for people with more skills in the lab and the > > > compensation > can > >be > > > better than a medical lab. > > > > > > mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion > > > about histotechnologist training. > > > > > > In regards to hospital histology labs in the USA: > > > > > > I don't want to start a firestorm of criticism here, but I think > > > there is a real danger that in Great Britain those technologists > >performing > > > in the histology laboratories may be over qualified.A few months > > > ago > > >here > >on > > > the Histonet, someone in the U.K. was bemoaning the lack of > histology > > > technologists in the U.K., and how short staffed they were. Was > > > that > >because > > > the bar had been set too high? Was the level of education required > too > >much > > > for the compensation received? Has a "closed shop" (to use a union > >term!) > >in > > > effect, been created? > > > Here in the United States there are hundreds of community type > > > hospitals and smaller medical centers that have histology > laboratories. > >In > > > those laboratories work histotechnologists doing a fine job with a > lot > >less > > > education than a fully qualified histotechnologist in the U.K.. At > the > >other > > > end of the spectrum are histology laboratories here doing more > advanced > > > procedures where degrees and extra training are required. > > > However, histotechnology training in the U.K. and USA are > > > inherently different. When I lived in the U.K. years and years ago > > > (and it may > have > > > changed now), all the hospitals were government owned, lab > > > personnel > got > >a > > > day off each week with pay to attend IMLT classes, and the classes > were > > > free. To work in a hospital laboratory you had to be "State > Registered". > > > In the USA hospitals are owned by all sorts of organizations. You > > > don't get "day release" to attend classes. If you do attend > > > classes, > > >money > > > has to come from somewhere to pay for it. Further you may live > >miles/hours > > > from the nearest college where you can attend classes.Since the > >different > > > organisations may be "for profit" or even "Non-profit", they want > > > to > > >save > > > money by not paying too higher salaries. Most often they pay what > other > > > hospitals in the area pay for the same type work. > > > The 50 states have different requirements for working in their > > > hospital laboratories. > > > Hospitals have coped with these differences by making histology > > > laboratories a separate section of the laboratory, with > > > technicians/technologists trained just to work in that lab. > > > I think a histotechnologist from the U.K. could easily find a > > > position in the USA, but they might feel underappreciated and > underpaid! > > > (maybe even under challanged!) > > > Well, that's my two bits worth! > > > > > > Mike Titford > > > USA Pathology > > > Mobile AL USA > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > > > > --------------------------------- Start your day with Yahoo! - > > > make it your home page > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@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 > > _________________________________________________________________ > Express yourself instantly with MSN Messenger! Download today - it's > FREE! > http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Mon Aug 8 13:17:43 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328CFD6@usca0082k08.labvision.apogent.com> Pam wrote: "We need better PR for the field as most people have not idea what a histologist is, much less does what we do. " You're right. It takes some extra work and no one is going to pay you for it, but it is fun and rewarding. Here are some things I do: Have a booth at career days for junior colleges (two-year degree "vocational" schools for the non-US Histonetters) and high schools. Give talks at the colleges and high schools in my area about our field (Teachers are desparate for these kinds of talks that will show students thay can use what they are learning). Sponsor interns at our company. Most of these are volunteer positions. (There are a LOT of high school and colledge students who are really interested in exploring the various opportunities. They are happy to work for free to get some experience. If you give them just a little training they can often find a job at their college working in a research lab. One word of advice: don't treat them like grunts to do only the dirty work.) Offer to write articles about the field for your local weekly newspaper - they will love you forever. I helped to get a sidebar about histotechnology into a nationally-used high school biology text book. The point is the "We" you mention is everyone who works in the field and is interested in seeing it advance. The NSH produces a brochure and a video, but no one will see it unless "we" show it to them. ASCP has some nice webpages showing the Histology field, but unless "we" point people to it, they won't see it. I encourage every histologist to do at least one thing each year to promote the field. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of mucram11@comcast.net Sent: Monday, August 08, 2005 10:51 AM To: Rittman, Barry R; histonet Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? I agree wholehaeartedly with Barry. We need better PR for the field as most people have not idea what a histologist is, much less does what we do. The brief acknowledgements on the crime shows and CSI saying well this is the histology reort or more often pathology report or send it to histology or pathology will not get us any recoginition. Everyone knows blood and body fluids go to a laboratory for testing few know about surgical specimens or what is done with them. The pahologist are still happy to hire anyone who says they want to learn histology and not worry about a degree because for too long we have allowed it and ASCP (pathologist) were getting off cheap. They don't want to pay a going rate for a field we have not stood and fought for in the past. Education has remained the most important thing for med tech and yet they are more automated today and less hands on than we will ever be. We need more schools and as Barry suggests even though we all hate the word standarization we will only grow and get recognition when the field is able to show we have testing and abilities we can prove. No one wants to pay the same for a person who learned on the job as for the person who has a degree. I know many histologist who were OJT and trust them with my specimens however, the times have changed and we need to change to grow starting with NSH and the members coming up with ways to let people know who we are and what we do. Sorry if this steps on some toes however my first years in histology were all OJT and I am proud of what I learned and that I had the chance learn more over the years. Pam Marcum -------------- Original message -------------- > The major problem with the situation in the USA re histotech that I > see > ... > There is a shortage of histotechs and instead of improving the salary > and career prospects many health organizations are hiring people who are > not trained and will work for much less, keeping the overall salary base > low. Cannot blame the people who are taking these jobs as they have to > have some work. > This situation will not change unless we have some attitude changes in > management and in the training that is available. > I personally would prefer the NSH to have its own testing and its own > testing centers, for all histotechs at a certain level to have to be > certified and for all individuals below this level to be in formal > training programs. This requires some sort of standardization for > training and testing across the country. While I hate the thought of the > federal government being in charge of anything, I think that this is the > only way in which respect for the profession and salaries will increase. > Finally we have a big public relations gap. Very few members of the > general public have any idea what histotechs do. > Barry > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Terry > Murphy > Sent: Saturday, August 06, 2005 7:34 AM > To: hawkmoon15@cox.net; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > HI everyone > > I feel the urge to join this discussion. I got into the field of > histology > 15 years ago and enjoyed the work and the concepts. I wanted to move up > in > the field ao I earned my HTL (ASCP) instead of the HT(ASCP) since I have > a > Bachelor's degree in Biology. That didn't get me any more respect or > money. > Still trying to improve my situation I earn a Master's degree in > Health > Administration and I once again my employment has not improved. As a > matter > of fact my employment situation has gotten worse since I earned a > Master's > degree. I had acheived a position as a supervisor only to be "run out > of > the lab" by an arrogant PA and, I can only assume, seasoned histotechs > that > fear change. > > Any way now I have been working as a traveling histotech. The pay is > better > than if I had a permanent job. The facility I am at now recently hired > a > person to be trained as a tech. This new employee has a GED not a high > school dipolma and he has no background in in science or medicine. He > is > doing good given his background but I find it kind of a insult to > educated > techs like myself that that someone without education or experience has > been > hired to do the same job that us techs have studied extensively. > > Now I am done traveling and am trying to get a histotech job close to > my > > home town so I can live in my house and more importantly live with my > wife. > I have applied for tech positions and either haven't heard from the > facility > except for a rejection letter. When I can get a hold of a hireing > supervisor I have been told that "this is a entry level position and it > doesn't pay well", "you are overqualified", and, this really surprised > me, > "your skill set will not go well in our laboratory". Someone on this > board > mentioned about being "black listed" I feel that I have now entered that > > list. Most likely I have more education than the people that I would > be > > reporting to and they probably feel threatened by me. Someone > suggested > > that I "dumb down" my resume and exclude some education and my > publication > but I do not like the idea of excluding accomplishments that I am proud > of. > > Lastly, as for wages, histotechs are grossly underpaid while > Pathologist > > Assistants are grossly overpaid!!!! > > Thanks for letting me vent. Sincerely, > > Another fustrated histotech. > > >From: "Sarah Jones" > >To: > >Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > >Date: Fri, 5 Aug 2005 16:12:29 -0700 > > > >I have to remind you here, Diana, that you are speaking for yourself > only > >with your statement about "in NA we tend to stay put". That may be > >for > > >some > >people. However, I have found throughout my ~40 year career in > >Histotechnology (especially in the last half of it!!) the only way to > get a > >decent raise is to pack up and move on. Unfortunately, here in NA our > >annual raises do not even keep up with the cost of living. In fact, it > is > >down-right terrible!! I have my HTL and got my CM recently, I am > >proud > to > >say. However, I haven't received any additional compensation for > either, > >and I doubt I ever will. (HTL for 24 yrs.) > > > >Which brings up something else: The ASCP used to be the American > Society > >of > >Clinical Pathologists. It is now the American Society of Clinical > >Pathology. What changed? I'm not sure. Does anyone know, except the > >'insiders', that is? I do have first-hand knowledge of what used to > be. > >The Pathologists ran it, and they had a lot to do with where > >Histotechs > >sat -- low on the rung of the Pathology Laboratory ladder for YEARS. > The > >Registry was in my home town in Muncie, Indiana, and my Mother was > >the > >secretary to the Registrar for many years. (This was all before it was > >centralized in Chicago. Prior to the relocation to Chicago, the > Registry > >was in the town where the Chairman of the Board was located. L. > >Montgomery, > >M.D., was the Chairman of the Board for ~30 years that I know of, and > thus > >was the Registry located in Muncie.) There are other things I could > tell > >you; but I was sworn to secrecy years ago, and I will honor that > pledge. > >Suffice it to say, the Pathologists wanted to keep the salaries of > >the > >Histotechs down, and did everything in their power to do so. 'Nuff > said! > > > >When I started into the field of Histotechnology at the University of > >Chicago back in 1965, the Techies associated with the ASCP were trying > to > >change the requirements for Histotechs, but to no avail. Not until > January > >of 2005 did the change they were attempting to accomplish way back > >then > >prevail. I didn't really understand what was happening then, but > today I > >do. There are MANY places here in the USA (where there is a severe > >shortage > >of Histotechs) where they will still hire techs that are not licensed. > In > >fact, there are places where a licensed Histologic Technician OR a > licensed > >Histotechnologist are black-listed by the other techs--most likely in > fear > >that their own incompetence might be discovered! > > > >The cost of living in certain areas of the country are no longer > >taken > into > >consideration in the way they were in the past when it comes to > >hiring > >techs. That also saddens me. How is it that they think I can live in > the > >most expensive county in California that pays the lowest wages of any > >county > >in California?? That is craziness in my book!!!! Yet they wonder why > they > >cannot find good Histotechs here in this county! GO FIGURE!! > > > >Anyway, I could go on and on, but I won't. Hope this sheds some light > on > >some things for some folks. Please address any personal questions to: > >hawkmoon15@cox.net > > > >Sarah > > > > > >----- Original Message ----- > >From: "Diana McCaig" > >To: > >Sent: Friday, August 05, 2005 10:41 AM > >Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > > > > > What are you all using to compare the wage scales. Biomedical > >scientists > >do > > > not get paid as well as we do in North America if you factor in > their > >cost > > > of living. I understand there is a recent trend to increase the > wages > >in > > > order to keep them on board as there is a lot of migration to > various > >labs > > > throughout a career where in NA we tend to stay put. > > > Diana McCaig, MLT > > > > > > > > > -----Original Message----- > > > From: Larry Woody [SMTP:slappycraw@yahoo.com] > > > Sent: Friday, August 05, 2005 1:35 PM > > > To: mtitford@aol.com; histonet@lists.utsouthwestern.edu > > > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > You may want to try some of the big biotech companies where there > > > is > > > a need for people with more skills in the lab and the compensation > can > >be > > > better than a medical lab. > > > > > > mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion > > > about histotechnologist training. > > > > > > In regards to hospital histology labs in the USA: > > > > > > I don't want to start a firestorm of criticism here, but I think > > > there is a real danger that in Great Britain those technologists > >performing > > > in the histology laboratories may be over qualified.A few months > > > ago > > >here > >on > > > the Histonet, someone in the U.K. was bemoaning the lack of > histology > > > technologists in the U.K., and how short staffed they were. Was > > > that > >because > > > the bar had been set too high? Was the level of education required > too > >much > > > for the compensation received? Has a "closed shop" (to use a union > >term!) > >in > > > effect, been created? > > > Here in the United States there are hundreds of community type > > > hospitals and smaller medical centers that have histology > laboratories. > >In > > > those laboratories work histotechnologists doing a fine job with a > lot > >less > > > education than a fully qualified histotechnologist in the U.K.. At > the > >other > > > end of the spectrum are histology laboratories here doing more > advanced > > > procedures where degrees and extra training are required. > > > However, histotechnology training in the U.K. and USA are inherently > > > different. When I lived in the U.K. years and years ago (and it may > have > > > changed now), all the hospitals were government owned, lab > > > personnel > got > >a > > > day off each week with pay to attend IMLT classes, and the classes > were > > > free. To work in a hospital laboratory you had to be "State > Registered". > > > In the USA hospitals are owned by all sorts of organizations. You > > > don't get "day release" to attend classes. If you do attend classes, > > >money > > > has to come from somewhere to pay for it. Further you may live > >miles/hours > > > from the nearest college where you can attend classes.Since the > >different > > > organisations may be "for profit" or even "Non-profit", they want > > > to > > >save > > > money by not paying too higher salaries. Most often they pay what > other > > > hospitals in the area pay for the same type work. > > > The 50 states have different requirements for working in their > > > hospital laboratories. > > > Hospitals have coped with these differences by making histology > > > laboratories a separate section of the laboratory, with > > > technicians/technologists trained just to work in that lab. > > > I think a histotechnologist from the U.K. could easily find a > > > position in the USA, but they might feel underappreciated and > underpaid! > > > (maybe even under challanged!) > > > Well, that's my two bits worth! > > > > > > Mike Titford > > > USA Pathology > > > Mobile AL USA > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > > > > --------------------------------- > > > Start your day with Yahoo! - make it your home page > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@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 > > _________________________________________________________________ > Express yourself instantly with MSN Messenger! Download today - it's > FREE! > http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rachael_Emerson <@t> URMC.Rochester.edu Mon Aug 8 13:34:37 2005 From: Rachael_Emerson <@t> URMC.Rochester.edu (Emerson, Rachael) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] whole mount IHC Message-ID: Hello. I am looking for advice on doing whole mount immunohistochemistry, without fixing the tissue in paraformaldehyde first, but using acetone, or possibly methanol, for fixation. I will be using mouse embryos. Any suggestions would be greatly appreciated. Thanks Rachael Rachael L. Emerson Center for Pediatric Biomedical Research University of Rochester Medical Center 575 Elmwood Avenue MRBX 1.11301 Rochester, NY 14642 Tel (585) 275-5073 Fax (585) 276-0232 From jamie.erickson <@t> abbott.com Mon Aug 8 14:07:02 2005 From: jamie.erickson <@t> abbott.com (Jamie E Erickson) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Tissues for HTL can anyone help. Message-ID: Hello All, I would like to know if anyone would have access to some tissues for HTL practical. Since I do Research I have been able to get Dog tissue that work but Ovary,Uterus, and Thyroid are hard to get that are in the right size. Unfortunately they won't accept my tiny mouse tissues. I am having trouble getting these 3 tissues that fit the size requirements. Any help would be greatly appreciated. Thanks _______________________________ Jamie Erickson Sr. Research Associate Department: DSMP Abbott Bioresearch Center 100 Research Drive Worcester, MA 01605-4341 508-688-3134 FAX: 508-793-4895 e-mail: jamie.erickson@abbott.com From mucram11 <@t> comcast.net Mon Aug 8 14:17:38 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: <080820051917.26150.42F7AFD0000ABDEE000066262200734364CECE030E9D0C9A03@comcast.net> Tim, I understand what you are doing and it great however, that was not my point. We need to get an understanding of histology and what is not just in schools but to ASCP (who as group has always under valued histology) and the public to understand what we do. Unitl the ASCP pathologist and NSH helps find ways to get the salaries and recognition we deserve and need it does not matter how many people we bring to field they will still be under paid and over worked. We need schools or programs to teach histology or the people you are talking to will have no place to go learn it as profession with a degree whether 2 year or four year. Until we have the places to get the education and the programs to expand our knowledge we have nothing offer given the new rules (which I applaud if we had the resources to fulfill it). Many places do not have schools and online can be great - you still need a place to practice and do the theory for yourself. Once you are working someplace at a low wage as an aid or OJY whlie you are getting certified is not being recoginized in many places with the monetary value it brings. If you worked cheap before a small increase is all that is offered, not a true wage comparable to a MT with certification. I have explained to so many people in the store, at the gym or just walking around what Histology means only because it was on a shirt I was wearing and they asked about what kind of history I taught. I have a nice way to explain and not offend anyone but I hate being asked if that is what a CSI does on the side. Pam -------------- Original message -------------- > Pam wrote: "We need better PR for the field as most people have not idea > what a histologist is, much less does what we do. " > > You're right. > > It takes some extra work and no one is going to pay you for it, but it is > fun and rewarding. > > Here are some things I do: > > Have a booth at career days for junior colleges (two-year degree > "vocational" schools for the non-US Histonetters) and high schools. > Give talks at the colleges and high schools in my area about our field > (Teachers are desparate for these kinds of talks that will show students > thay can use what they are learning). > Sponsor interns at our company. Most of these are volunteer positions. > (There are a LOT of high school and colledge students who are really > interested in exploring the various opportunities. They are happy to work > for free to get some experience. If you give them just a little training > they can often find a job at their college working in a research lab. One > word of advice: don't treat them like grunts to do only the dirty work.) > Offer to write articles about the field for your local weekly newspaper - > they will love you forever. > I helped to get a sidebar about histotechnology into a nationally-used high > school biology text book. > > > The point is the "We" you mention is everyone who works in the field and is > interested in seeing it advance. The NSH produces a brochure and a video, > but no one will see it unless "we" show it to them. ASCP has some nice > webpages showing the Histology field, but unless "we" point people to it, > they won't see it. > > I encourage every histologist to do at least one thing each year to promote > the field. > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > mucram11@comcast.net > Sent: Monday, August 08, 2005 10:51 AM > To: Rittman, Barry R; histonet > Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? > > > I agree wholehaeartedly with Barry. We need better PR for the field as most > people have not idea what a histologist is, much less does what we do. The > brief acknowledgements on the crime shows and CSI saying well this is the > histology reort or more often pathology report or send it to histology or > pathology will not get us any recoginition. Everyone knows blood and body > fluids go to a laboratory for testing few know about surgical specimens or > what is done with them. > > The pahologist are still happy to hire anyone who says they want to learn > histology and not worry about a degree because for too long we have allowed > it and ASCP (pathologist) were getting off cheap. They don't want to pay a > going rate for a field we have not stood and fought for in the past. > Education has remained the most important thing for med tech and yet they > are more automated today and less hands on than we will ever be. We need > more schools and as Barry suggests even though we all hate the word > standarization we will only grow and get recognition when the field is able > to show we have testing and abilities we can prove. > > No one wants to pay the same for a person who learned on the job as for the > person who has a degree. I know many histologist who were OJT and trust > them with my specimens however, the times have changed and we need to change > to grow starting with NSH and the members coming up with ways to let people > know who we are and what we do. Sorry if this steps on some toes however > my first years in histology were all OJT and I am proud of what I learned > and that I had the chance learn more over the years. > > Pam Marcum > -------------- Original message -------------- > > > The major problem with the situation in the USA re histotech that I > > see > > ... > > There is a shortage of histotechs and instead of improving the salary > > and career prospects many health organizations are hiring people who are > > not trained and will work for much less, keeping the overall salary base > > low. Cannot blame the people who are taking these jobs as they have to > > have some work. > > This situation will not change unless we have some attitude changes in > > management and in the training that is available. > > I personally would prefer the NSH to have its own testing and its own > > testing centers, for all histotechs at a certain level to have to be > > certified and for all individuals below this level to be in formal > > training programs. This requires some sort of standardization for > > training and testing across the country. While I hate the thought of the > > federal government being in charge of anything, I think that this is the > > only way in which respect for the profession and salaries will increase. > > Finally we have a big public relations gap. Very few members of the > > general public have any idea what histotechs do. > > Barry > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Terry > > Murphy > > Sent: Saturday, August 06, 2005 7:34 AM > > To: hawkmoon15@cox.net; histonet@lists.utsouthwestern.edu > > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > > > HI everyone > > > > I feel the urge to join this discussion. I got into the field of > > histology > > 15 years ago and enjoyed the work and the concepts. I wanted to move up > > in > > the field ao I earned my HTL (ASCP) instead of the HT(ASCP) since I have > > a > > Bachelor's degree in Biology. That didn't get me any more respect or > > money. > > Still trying to improve my situation I earn a Master's degree in > > Health > > Administration and I once again my employment has not improved. As a > > matter > > of fact my employment situation has gotten worse since I earned a > > Master's > > degree. I had acheived a position as a supervisor only to be "run out > > of > > the lab" by an arrogant PA and, I can only assume, seasoned histotechs > > that > > fear change. > > > > Any way now I have been working as a traveling histotech. The pay is > > better > > than if I had a permanent job. The facility I am at now recently hired > > a > > person to be trained as a tech. This new employee has a GED not a high > > school dipolma and he has no background in in science or medicine. He > > is > > doing good given his background but I find it kind of a insult to > > educated > > techs like myself that that someone without education or experience has > > been > > hired to do the same job that us techs have studied extensively. > > > > Now I am done traveling and am trying to get a histotech job close to > > my > > > > home town so I can live in my house and more importantly live with my > > wife. > > I have applied for tech positions and either haven't heard from the > > facility > > except for a rejection letter. When I can get a hold of a hireing > > supervisor I have been told that "this is a entry level position and it > > doesn't pay well", "you are overqualified", and, this really surprised > > me, > > "your skill set will not go well in our laboratory". Someone on this > > board > > mentioned about being "black listed" I feel that I have now entered that > > > > list. Most likely I have more education than the people that I would > > be > > > > reporting to and they probably feel threatened by me. Someone > > suggested > > > > that I "dumb down" my resume and exclude some education and my > > publication > > but I do not like the idea of excluding accomplishments that I am proud > > of. > > > > Lastly, as for wages, histotechs are grossly underpaid while > > Pathologist > > > > Assistants are grossly overpaid!!!! > > > > Thanks for letting me vent. Sincerely, > > > > Another fustrated histotech. > > > > >From: "Sarah Jones" > > >To: > > >Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > >Date: Fri, 5 Aug 2005 16:12:29 -0700 > > > > > >I have to remind you here, Diana, that you are speaking for yourself > > only > > >with your statement about "in NA we tend to stay put". That may be > > >for > > > > >some > > >people. However, I have found throughout my ~40 year career in > > >Histotechnology (especially in the last half of it!!) the only way to > > get a > > >decent raise is to pack up and move on. Unfortunately, here in NA our > > >annual raises do not even keep up with the cost of living. In fact, it > > is > > >down-right terrible!! I have my HTL and got my CM recently, I am > > >proud > > to > > >say. However, I haven't received any additional compensation for > > either, > > >and I doubt I ever will. (HTL for 24 yrs.) > > > > > >Which brings up something else: The ASCP used to be the American > > Society > > >of > > >Clinical Pathologists. It is now the American Society of Clinical > > >Pathology. What changed? I'm not sure. Does anyone know, except the > > >'insiders', that is? I do have first-hand knowledge of what used to > > be. > > >The Pathologists ran it, and they had a lot to do with where > > >Histotechs > > >sat -- low on the rung of the Pathology Laboratory ladder for YEARS. > > The > > >Registry was in my home town in Muncie, Indiana, and my Mother was > > >the > > >secretary to the Registrar for many years. (This was all before it was > > >centralized in Chicago. Prior to the relocation to Chicago, the > > Registry > > >was in the town where the Chairman of the Board was located. L. > > >Montgomery, > > >M.D., was the Chairman of the Board for ~30 years that I know of, and > > thus > > >was the Registry located in Muncie.) There are other things I could > > tell > > >you; but I was sworn to secrecy years ago, and I will honor that > > pledge. > > >Suffice it to say, the Pathologists wanted to keep the salaries of > > >the > > >Histotechs down, and did everything in their power to do so. 'Nuff > > said! > > > > > >When I started into the field of Histotechnology at the University of > > >Chicago back in 1965, the Techies associated with the ASCP were trying > > to > > >change the requirements for Histotechs, but to no avail. Not until > > January > > >of 2005 did the change they were attempting to accomplish way back > > >then > > >prevail. I didn't really understand what was happening then, but > > today I > > >do. There are MANY places here in the USA (where there is a severe > > >shortage > > >of Histotechs) where they will still hire techs that are not licensed. > > In > > >fact, there are places where a licensed Histologic Technician OR a > > licensed > > >Histotechnologist are black-listed by the other techs--most likely in > > fear > > >that their own incompetence might be discovered! > > > > > >The cost of living in certain areas of the country are no longer > > >taken > > into > > >consideration in the way they were in the past when it comes to > > >hiring > > >techs. That also saddens me. How is it that they think I can live in > > the > > >most expensive county in California that pays the lowest wages of any > > >county > > >in California?? That is craziness in my book!!!! Yet they wonder why > > they > > >cannot find good Histotechs here in this county! GO FIGURE!! > > > > > >Anyway, I could go on and on, but I won't. Hope this sheds some light > > on > > >some things for some folks. Please address any personal questions to: > > >hawkmoon15@cox.net > > > > > >Sarah > > > > > > > > >----- Original Message ----- > > >From: "Diana McCaig" > > >To: > > >Sent: Friday, August 05, 2005 10:41 AM > > >Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > > > > > > > > > What are you all using to compare the wage scales. Biomedical > > >scientists > > >do > > > > not get paid as well as we do in North America if you factor in > > their > > >cost > > > > of living. I understand there is a recent trend to increase the > > wages > > >in > > > > order to keep them on board as there is a lot of migration to > > various > > >labs > > > > throughout a career where in NA we tend to stay put. > > > > Diana McCaig, MLT > > > > > > > > > > > > -----Original Message----- > > > > From: Larry Woody [SMTP:slappycraw@yahoo.com] > > > > Sent: Friday, August 05, 2005 1:35 PM > > > > To: mtitford@aol.com; histonet@lists.utsouthwestern.edu > > > > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > > > You may want to try some of the big biotech companies where there > > > > is > > > > a need for people with more skills in the lab and the compensation > > can > > >be > > > > better than a medical lab. > > > > > > > > mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion > > > > about histotechnologist training. > > > > > > > > In regards to hospital histology labs in the USA: > > > > > > > > I don't want to start a firestorm of criticism here, but I think > > > > there is a real danger that in Great Britain those technologists > > >performing > > > > in the histology laboratories may be over qualified.A few months > > > > ago > > > > >here > > >on > > > > the Histonet, someone in the U.K. was bemoaning the lack of > > histology > > > > technologists in the U.K., and how short staffed they were. Was > > > > that > > >because > > > > the bar had been set too high? Was the level of education required > > too > > >much > > > > for the compensation received? Has a "closed shop" (to use a union > > >term!) > > >in > > > > effect, been created? > > > > Here in the United States there are hundreds of community type > > > > hospitals and smaller medical centers that have histology > > laboratories. > > >In > > > > those laboratories work histotechnologists doing a fine job with a > > lot > > >less > > > > education than a fully qualified histotechnologist in the U.K.. At > > the > > >other > > > > end of the spectrum are histology laboratories here doing more > > advanced > > > > procedures where degrees and extra training are required. > > > > However, histotechnology training in the U.K. and USA are inherently > > > > different. When I lived in the U.K. years and years ago (and it may > > have > > > > changed now), all the hospitals were government owned, lab > > > > personnel > > got > > >a > > > > day off each week with pay to attend IMLT classes, and the classes > > were > > > > free. To work in a hospital laboratory you had to be "State > > Registered". > > > > In the USA hospitals are owned by all sorts of organizations. You > > > > don't get "day release" to attend classes. If you do attend classes, > > > > >money > > > > has to come from somewhere to pay for it. Further you may live > > >miles/hours > > > > from the nearest college where you can attend classes.Since the > > >different > > > > organisations may be "for profit" or even "Non-profit", they want > > > > to > > > > >save > > > > money by not paying too higher salaries. Most often they pay what > > other > > > > hospitals in the area pay for the same type work. > > > > The 50 states have different requirements for working in their > > > > hospital laboratories. > > > > Hospitals have coped with these differences by making histology > > > > laboratories a separate section of the laboratory, with > > > > technicians/technologists trained just to work in that lab. > > > > I think a histotechnologist from the U.K. could easily find a > > > > position in the USA, but they might feel underappreciated and > > underpaid! > > > > (maybe even under challanged!) > > > > Well, that's my two bits worth! > > > > > > > > Mike Titford > > > > USA Pathology > > > > Mobile AL USA > > > > _______________________________________________ > > > > Histonet mailing list > > > > Histonet@lists.utsouthwestern.edu > > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > > > > > > > > --------------------------------- > > > > Start your day with Yahoo! - make it your home page > > > > _______________________________________________ > > > > Histonet mailing list > > > > Histonet@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 > > > > _________________________________________________________________ > > Express yourself instantly with MSN Messenger! Download today - it's > > FREE! > > http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From maria <@t> ski.org Mon Aug 8 16:19:27 2005 From: maria <@t> ski.org (Maria Mejia) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Cresyl Violet stain for bird brains References: Message-ID: <42F7CC5F.8090405@ski.org> Hello Brandon, The following cresyl violet acetate stain is one I have used on several different animal species . What I found is that the pH is very critical in making this stain. If it's not right will result in NO staining! CV stock solution: (I purchase my crysyl violet acetate dye from Fluka #61123. You can order this dye from Sigma 800-325-3010 or see website). 0.9gm Dist H20 - 75ml Mix well in a brown bottle. I let it mix- overnight Then make up the acetate buffer A & B solutions: Sol A: glacial acetic acid- 3ml DH20 - 250ml Mix well. Sol B: sodium acetate-0.6gm DH20 - 75ml Working Sol. of CV stain Sol. A - 237ml Sol. B - 63ml Add CV stock from 50ml to 70ml (I sometimes use 50ml to 60ml) Mix well in brown bottle and pH 5.0 - this is very important. Filter before use once a day & check pH too!!! This dye solution is stable & can keep at RT . Staining w/CV: I usually stain 1 or 2 slides for testing. Defatt in xylene (to remove lipid membrane from cells) - 1 change 10 dips - 20mins. hydrate in 100% (ethyl or reagent) alcohol) - 3 changes - 10 dips ea. - 4 mins ea. 95% alcohol - 1 change 10 dips - 4mins. 70% alcohol - 1 change 10 dips - 4mins. Cresyl violet acetate stain - 10 slow dips - stain 30 sec - 2 mins. **remember fresh stain is a bit stronger. Try a 1 minute staining time. dehydrate: 70% alcohol - 10 dips - 1min. (sometimes might need to do this step for 1.5mins). (during the staining procedure this alcohol must be changed so the alcohol looks clear and not blue. If blue color, it will stain your sections further). 95% alcohol - 10 dips - 1 min. 100% alcohol - 3 changes - 10 dips ea. + 4 mins ea. **should see a nice clear difference between the grey & white matter. xylene - 3 changes - 10 dips ea. + 5 mins ea. coverslip. I hope this helps. Yours Maria Bartola Mejia neurohistologist Smith-Kettlewell Eye Res. Inst. San Francisco, Ca Email: maria@ski.org Brandon Munk wrote: >I am having trouble obtaining an acceptable Nissl stain of passerine brains >using cresyl violet. The birds were transcardially perfused, brains removed, >postfixed and cryoprotected before sectioning at 40 um on a sledge >microtome. The protocol I am using was originally used for rat brains and is >as follow: > > ddH20 (few dips) > cresyl violet (5-10 min) > ddH20 (rinse excess stain) > 70% etOH (few dips) > 95% etOH (few dips) > 95% etOH + acetic acid (few dips) > ddH20 (few dips) > 95% etOH (few dips) > 100% etOH (few dips) > repeat previous steps as necessary > Xylene (one dip) > coverslip > >I am unsure as to the concentration of cresyl violet I am using and it has >sat in a fridge for >2 yrs (although I have been assured that the age should >not be a problem). I have tried playing with the protocol quite a bit, >varying time in cresyl violet or how many times I dehydrate and >differentiate. I am getting near my wits end and still cannot get a good >stain. All my sections look as if they are washed out and I often have some >stain 'smeared' under the cover slip after dipping in xylene. > >Can anyone help me? Thanks in advance! > >Brandon >_____ >Brandon Munk >Department of Zoology and Physiology >University of Wyoming >PO Box 3166 >Laramie, WY 82071 >_____ >bmunk@uwyo.edu > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From cwscouten <@t> myneurolab.com Mon Aug 8 16:23:47 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] fixation via perfusion Message-ID: <5784D843593D874C93E9BADCB87342AB9164DB@tpiserver03.Coretech-holdings.com> See the following link for a discussion of perfusion. Consider montitoring pressure, not flow rate, see the link to see why, and I agree that the most common error is pressure too low. The vascular system can clearly handle at least normal cardiac pressure levels, which way exceed typical gravity flow or common perestaltic settings. http://www.myneurolab.com/myneurolab/mnl_articles.asp?ProductID=471001&idsubcategory=21&idproduct=471001&catdesc=Histology+Equipment&subcatdesc=Sacrifice+Equipment&AddInfoStr=2 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 Geoff McAuliffe Sent: Monday, August 08, 2005 12:22 PM To: Wisam Barkho Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] fixation via perfusion Hi Wisam: I don't have the exact answer you are looking for but I can provide some direction. This information was probably worked out in the late 1950's and/or early 1960's as perfusion fixation emerged as the way to fix for electron microscopy. I would suggest looking in Hayat's "Principles and Techniques for Electron Microscopy" or Lillie and Fullmer's "Histological Technique and Practical Histochemistry" as places to look for the original references. In my experience, most people use too small a canula and/or too low a flow rate for good fixation. The rate of flow you use should approximate the cardiac output of the animal in question. As far as volume goes, I usually use a 3 times the weight of the animal if I am perfusing the whole body, less if you clamp off the lower body and perfuse just the head. Good luck! Geoff Wisam Barkho wrote: >Hello, > > > >We are doing stereotaxic surgeries on rats in my lab. After 2 weeks, we >fixate the brain via perfusion using 4% PFA (after vascular rinse) and >then immerse the brains in 4% PFA overnight. My question is where can I >find information on pump flow rate and PFA volume to use during fixation? Thanks. > > > >Wisam Barkho > >daufoi@msn.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 ********************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Mon Aug 8 17:03:33 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] RE: A question to the Histonet Webmaster Message-ID: <5784D843593D874C93E9BADCB87342AB9164DE@tpiserver03.Coretech-holdings.com> The histonet is a listserve. You should be getting email for histonet inquiries. Are you? If so, you are subscribed. Histonet.org if not the histonet. If you have been to the subscription website, at http://lists.utsouthwestern.edu/mailman/listinfo/histonet and subscribed, you are on the list. You may not log in unless you are linda margraf, you are not allowed to get access to the individual names on the list. To use the histonet, just send an email to histonet@lists.utsouthwestern.edu 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: Doug Martin Sent: Saturday, August 06, 2005 9:05 AM To: Charles Scouten Subject: FW: A question to the Histonet Webmaster -----Original Message----- From: webmaster@histonet.org [mailto:webmaster@histonet.org] Sent: Saturday, August 06, 2005 3:23 AM To: webmaster Subject: A question to the Histonet Webmaster textfield = lscoomer@earthlink.net textfield2 = Linda Coomer textfield3 = I have followed all intructions on subscribing to histonet,how-ever I cannot log on. Please help, I have received my confirmation number an entered it a number of times. No Luck Submit = Email From Kemlo.Rogerson <@t> elht.nhs.uk Tue Aug 9 02:05:59 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] "Brit" Hitotech/Firestorm? Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4D7@elht-exch1.xelht.nhs.uk> Did that myself, both in London and in East Lancs; the usual response? I want to be a Forensic Scientist like on the Telly; we have a lot of Forensic Scientist Telly Programmes. I want to be a Doctor, can I come and see want the Labs do? Then you spot the withdrawn student with acne and shunned by his mates; then you know you have a Biomedical Scientist/ HistoTech in the making. Billy no-mates who doesn't interact with people (thus cannot be a Doctor), is not motivated by money (thus cannot be a Doctor), Doesn't think the world revolves around him (thus cannot be a Doctor) and doesn't like the sight of blood (can't be a Forensic Scientist). Get him/ her, cherish him/ her and get him/ her up to be a MSc postgraduate; yes I know he/ she leaves to be a Doctor. The above is my views only and are the amused ramblings of a cynical old man. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: 08 August 2005 19:18 To: 'mucram11@comcast.net'; Rittman, Barry R; histonet Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? Pam wrote: "We need better PR for the field as most people have not idea what a histologist is, much less does what we do. " You're right. It takes some extra work and no one is going to pay you for it, but it is fun and rewarding. Here are some things I do: Have a booth at career days for junior colleges (two-year degree "vocational" schools for the non-US Histonetters) and high schools. Give talks at the colleges and high schools in my area about our field (Teachers are desparate for these kinds of talks that will show students thay can use what they are learning). Sponsor interns at our company. Most of these are volunteer positions. (There are a LOT of high school and colledge students who are really interested in exploring the various opportunities. They are happy to work for free to get some experience. If you give them just a little training they can often find a job at their college working in a research lab. One word of advice: don't treat them like grunts to do only the dirty work.) Offer to write articles about the field for your local weekly newspaper - they will love you forever. I helped to get a sidebar about histotechnology into a nationally-used high school biology text book. The point is the "We" you mention is everyone who works in the field and is interested in seeing it advance. The NSH produces a brochure and a video, but no one will see it unless "we" show it to them. ASCP has some nice webpages showing the Histology field, but unless "we" point people to it, they won't see it. I encourage every histologist to do at least one thing each year to promote the field. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of mucram11@comcast.net Sent: Monday, August 08, 2005 10:51 AM To: Rittman, Barry R; histonet Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? I agree wholehaeartedly with Barry. We need better PR for the field as most people have not idea what a histologist is, much less does what we do. The brief acknowledgements on the crime shows and CSI saying well this is the histology reort or more often pathology report or send it to histology or pathology will not get us any recoginition. Everyone knows blood and body fluids go to a laboratory for testing few know about surgical specimens or what is done with them. The pahologist are still happy to hire anyone who says they want to learn histology and not worry about a degree because for too long we have allowed it and ASCP (pathologist) were getting off cheap. They don't want to pay a going rate for a field we have not stood and fought for in the past. Education has remained the most important thing for med tech and yet they are more automated today and less hands on than we will ever be. We need more schools and as Barry suggests even though we all hate the word standarization we will only grow and get recognition when the field is able to show we have testing and abilities we can prove. No one wants to pay the same for a person who learned on the job as for the person who has a degree. I know many histologist who were OJT and trust them with my specimens however, the times have changed and we need to change to grow starting with NSH and the members coming up with ways to let people know who we are and what we do. Sorry if this steps on some toes however my first years in histology were all OJT and I am proud of what I learned and that I had the chance learn more over the years. Pam Marcum -------------- Original message -------------- > The major problem with the situation in the USA re histotech that I > see > ... > There is a shortage of histotechs and instead of improving the salary > and career prospects many health organizations are hiring people who are > not trained and will work for much less, keeping the overall salary base > low. Cannot blame the people who are taking these jobs as they have to > have some work. > This situation will not change unless we have some attitude changes in > management and in the training that is available. > I personally would prefer the NSH to have its own testing and its own > testing centers, for all histotechs at a certain level to have to be > certified and for all individuals below this level to be in formal > training programs. This requires some sort of standardization for > training and testing across the country. While I hate the thought of the > federal government being in charge of anything, I think that this is the > only way in which respect for the profession and salaries will increase. > Finally we have a big public relations gap. Very few members of the > general public have any idea what histotechs do. > Barry > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Terry > Murphy > Sent: Saturday, August 06, 2005 7:34 AM > To: hawkmoon15@cox.net; histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > HI everyone > > I feel the urge to join this discussion. I got into the field of > histology > 15 years ago and enjoyed the work and the concepts. I wanted to move up > in > the field ao I earned my HTL (ASCP) instead of the HT(ASCP) since I have > a > Bachelor's degree in Biology. That didn't get me any more respect or > money. > Still trying to improve my situation I earn a Master's degree in > Health > Administration and I once again my employment has not improved. As a > matter > of fact my employment situation has gotten worse since I earned a > Master's > degree. I had acheived a position as a supervisor only to be "run out > of > the lab" by an arrogant PA and, I can only assume, seasoned histotechs > that > fear change. > > Any way now I have been working as a traveling histotech. The pay is > better > than if I had a permanent job. The facility I am at now recently hired > a > person to be trained as a tech. This new employee has a GED not a high > school dipolma and he has no background in in science or medicine. He > is > doing good given his background but I find it kind of a insult to > educated > techs like myself that that someone without education or experience has > been > hired to do the same job that us techs have studied extensively. > > Now I am done traveling and am trying to get a histotech job close to > my > > home town so I can live in my house and more importantly live with my > wife. > I have applied for tech positions and either haven't heard from the > facility > except for a rejection letter. When I can get a hold of a hireing > supervisor I have been told that "this is a entry level position and it > doesn't pay well", "you are overqualified", and, this really surprised > me, > "your skill set will not go well in our laboratory". Someone on this > board > mentioned about being "black listed" I feel that I have now entered that > > list. Most likely I have more education than the people that I would > be > > reporting to and they probably feel threatened by me. Someone > suggested > > that I "dumb down" my resume and exclude some education and my > publication > but I do not like the idea of excluding accomplishments that I am proud > of. > > Lastly, as for wages, histotechs are grossly underpaid while > Pathologist > > Assistants are grossly overpaid!!!! > > Thanks for letting me vent. Sincerely, > > Another fustrated histotech. > > >From: "Sarah Jones" > >To: > >Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > >Date: Fri, 5 Aug 2005 16:12:29 -0700 > > > >I have to remind you here, Diana, that you are speaking for yourself > only > >with your statement about "in NA we tend to stay put". That may be > >for > > >some > >people. However, I have found throughout my ~40 year career in > >Histotechnology (especially in the last half of it!!) the only way to > get a > >decent raise is to pack up and move on. Unfortunately, here in NA our > >annual raises do not even keep up with the cost of living. In fact, it > is > >down-right terrible!! I have my HTL and got my CM recently, I am > >proud > to > >say. However, I haven't received any additional compensation for > either, > >and I doubt I ever will. (HTL for 24 yrs.) > > > >Which brings up something else: The ASCP used to be the American > Society > >of > >Clinical Pathologists. It is now the American Society of Clinical > >Pathology. What changed? I'm not sure. Does anyone know, except the > >'insiders', that is? I do have first-hand knowledge of what used to > be. > >The Pathologists ran it, and they had a lot to do with where > >Histotechs > >sat -- low on the rung of the Pathology Laboratory ladder for YEARS. > The > >Registry was in my home town in Muncie, Indiana, and my Mother was > >the > >secretary to the Registrar for many years. (This was all before it was > >centralized in Chicago. Prior to the relocation to Chicago, the > Registry > >was in the town where the Chairman of the Board was located. L. > >Montgomery, > >M.D., was the Chairman of the Board for ~30 years that I know of, and > thus > >was the Registry located in Muncie.) There are other things I could > tell > >you; but I was sworn to secrecy years ago, and I will honor that > pledge. > >Suffice it to say, the Pathologists wanted to keep the salaries of > >the > >Histotechs down, and did everything in their power to do so. 'Nuff > said! > > > >When I started into the field of Histotechnology at the University of > >Chicago back in 1965, the Techies associated with the ASCP were trying > to > >change the requirements for Histotechs, but to no avail. Not until > January > >of 2005 did the change they were attempting to accomplish way back > >then > >prevail. I didn't really understand what was happening then, but > today I > >do. There are MANY places here in the USA (where there is a severe > >shortage > >of Histotechs) where they will still hire techs that are not licensed. > In > >fact, there are places where a licensed Histologic Technician OR a > licensed > >Histotechnologist are black-listed by the other techs--most likely in > fear > >that their own incompetence might be discovered! > > > >The cost of living in certain areas of the country are no longer > >taken > into > >consideration in the way they were in the past when it comes to > >hiring > >techs. That also saddens me. How is it that they think I can live in > the > >most expensive county in California that pays the lowest wages of any > >county > >in California?? That is craziness in my book!!!! Yet they wonder why > they > >cannot find good Histotechs here in this county! GO FIGURE!! > > > >Anyway, I could go on and on, but I won't. Hope this sheds some light > on > >some things for some folks. Please address any personal questions to: > >hawkmoon15@cox.net > > > >Sarah > > > > > >----- Original Message ----- > >From: "Diana McCaig" > >To: > >Sent: Friday, August 05, 2005 10:41 AM > >Subject: RE: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > > > > > What are you all using to compare the wage scales. Biomedical > >scientists > >do > > > not get paid as well as we do in North America if you factor in > their > >cost > > > of living. I understand there is a recent trend to increase the > wages > >in > > > order to keep them on board as there is a lot of migration to > various > >labs > > > throughout a career where in NA we tend to stay put. > > > Diana McCaig, MLT > > > > > > > > > -----Original Message----- > > > From: Larry Woody [SMTP:slappycraw@yahoo.com] > > > Sent: Friday, August 05, 2005 1:35 PM > > > To: mtitford@aol.com; histonet@lists.utsouthwestern.edu > > > Subject: Re: [Histonet] "Brit" Hitotech/Firestorm? > > > > > > You may want to try some of the big biotech companies where there > > > is > > > a need for people with more skills in the lab and the compensation > can > >be > > > better than a medical lab. > > > > > > mtitford@aol.com wrote:Rogerson Kemlo carries on the discussion > > > about histotechnologist training. > > > > > > In regards to hospital histology labs in the USA: > > > > > > I don't want to start a firestorm of criticism here, but I think > > > there is a real danger that in Great Britain those technologists > >performing > > > in the histology laboratories may be over qualified.A few months > > > ago > > >here > >on > > > the Histonet, someone in the U.K. was bemoaning the lack of > histology > > > technologists in the U.K., and how short staffed they were. Was > > > that > >because > > > the bar had been set too high? Was the level of education required > too > >much > > > for the compensation received? Has a "closed shop" (to use a union > >term!) > >in > > > effect, been created? > > > Here in the United States there are hundreds of community type > > > hospitals and smaller medical centers that have histology > laboratories. > >In > > > those laboratories work histotechnologists doing a fine job with a > lot > >less > > > education than a fully qualified histotechnologist in the U.K.. At > the > >other > > > end of the spectrum are histology laboratories here doing more > advanced > > > procedures where degrees and extra training are required. > > > However, histotechnology training in the U.K. and USA are inherently > > > different. When I lived in the U.K. years and years ago (and it may > have > > > changed now), all the hospitals were government owned, lab > > > personnel > got > >a > > > day off each week with pay to attend IMLT classes, and the classes > were > > > free. To work in a hospital laboratory you had to be "State > Registered". > > > In the USA hospitals are owned by all sorts of organizations. You > > > don't get "day release" to attend classes. If you do attend classes, > > >money > > > has to come from somewhere to pay for it. Further you may live > >miles/hours > > > from the nearest college where you can attend classes.Since the > >different > > > organisations may be "for profit" or even "Non-profit", they want > > > to > > >save > > > money by not paying too higher salaries. Most often they pay what > other > > > hospitals in the area pay for the same type work. > > > The 50 states have different requirements for working in their > > > hospital laboratories. > > > Hospitals have coped with these differences by making histology > > > laboratories a separate section of the laboratory, with > > > technicians/technologists trained just to work in that lab. > > > I think a histotechnologist from the U.K. could easily find a > > > position in the USA, but they might feel underappreciated and > underpaid! > > > (maybe even under challanged!) > > > Well, that's my two bits worth! > > > > > > Mike Titford > > > USA Pathology > > > Mobile AL USA > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > > > > --------------------------------- > > > Start your day with Yahoo! - make it your home page > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@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 > > _________________________________________________________________ > Express yourself instantly with MSN Messenger! Download today - it's > FREE! > http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From abee <@t> pml.ac.uk Tue Aug 9 09:30:10 2005 From: abee <@t> pml.ac.uk (abee@pml.ac.uk) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Methacrylate cuttting Message-ID: <000001c59cee$d9a66f40$21a2abc0@npm.ac.uk> Dear all, HELP!! To all of you this will probably be such an easy solution. My boss is on annual leave for 3 weeks and has left me the task of practising cutting methacrylate sections at 3microns. There probably wouldn't have been a problem but I have never cut them before. I have been able to cut the glass knives, the microtome is set up, but every time I go to cut, the section just ruffles up. I have immersed the slides in water so that the sections are being placed onto a wet surface. I have looked up on the web for protocols, but they are all for the process of blocking up or staining. Please can someone help!!! Regards, Amanda From Xorren <@t> aol.com Tue Aug 9 10:42:44 2005 From: Xorren <@t> aol.com (Xorren@aol.com) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] NY histology jobs anyone? Message-ID: <1fa.e3be8d0.302a28f4@aol.com> Hello Histonetters! I am a registered tech for 13 years and wonder if anyone knows of any positions available In NYC or preferably Long Island? I would appreciate any info! thanks in advance ! HL From RCHIOVETTI <@t> aol.com Tue Aug 9 11:07:10 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Methacrylate cuttting Message-ID: <12e.6399e0a1.302a2eae@aol.com> In a message dated 8/9/2005 7:31:29 AM US Mountain Standard Time, abee@pml.ac.uk writes: > HELP!! To all of you this will probably be such an easy solution. My boss > is > on annual leave for 3 weeks and has left me the task of practising cutting > methacrylate sections at 3microns. > > Amanda, If you are using glass knives, are they the triangular type of knives, or the longer type of "Ralph" knives? Are you using an ultramicrotome? Or are you using a glass knife adapter on a regular rotary microtome? And can you put a "boat" or a "trough" on the knife so it can hold a little water? Methacrylate tends to be a little soft, and it's not uncommon to see the sections crumple and stick to the knife edge. You could probably get better results if you could use the sectioning techniques that are used for electron microscopy. These involve bringing water up to the knife edge, so the sections float onto the water surface as they are cut. You can then retrieve the sections with a small wire loop or a hair mounted on a stick, and transfer them to a drop of water on a glass slide. Also, using water during sectioning will help the sections "relax" and flatten out. A lot of the methacrylates are hydrophilic, and you can actually watch them expand slightly as they come off the knife edge and float on the water surface. If you were cutting slightly thicker, say around 5-8 microns, you might be able to *carefully* use a pair of fine-tip watchmaker's forceps or a hair and gently guide the sections down the knife as they are cut. But this may not work at 3 microns thickness. This may have been discussed previously on Histonet...I can'r remember for sure. You can go to: and try searching the Histonet archives using words like: sectioning methacrylate Hope this helps! Cheers, Bob Robert (Bob) Chiovetti, Ph.D. The Microscope Works Arizona's Microscopy Resource 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From Jacqueline.Malam <@t> rli.mbht.nhs.uk Tue Aug 9 11:08:14 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] RE: Neg controls Message-ID: Check you are using the correct sera for your negative controls - should be the "Ig fraction" sera (the "whole sera" is used for the blocking stage). Your sera could be too low a dilution - you need to dilute it to the same protein concentration as the diluted primary. There is a formula: If dilution of primary=A and total protein concentration of primary =B And total protein concentration of neg. sera=C you divide A by B and multiply by C Then you just treat the slide to the same ag. retrieval process etc as the primary. You do end up with a lot of neg. control slides (no retrieval, enzyme, pressure cook, microwave, pressure cook and enzyme) both for polys and monos. We used to do the lot but now we are fully automated and there is only one poly and mono sera available on their commercial list so, based on the theory that if it's ok with them to stick with that, when we make our own antisera preps, we dilute both rabbit and mouse Ig sera 1/1,000. It's an arbitrary figure based on the recommended dilution for mouse sera from Dako years ago. If there are any problems (and there haven't been yet), we could always repeat with a "corrected for total protein concentration" dilution. Good luck! Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From ajennings <@t> unmc.edu Tue Aug 9 11:56:38 2005 From: ajennings <@t> unmc.edu (Anita Jennings) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Cryojane/gentle jane Message-ID: Steve I purchased what I believe was one of the first cryojane apparatus from Instrumedics when I ran the Histology Core Facility at Mayo Clinic in Scottsdale AZ. It took some getting use to but it made some beautiful frozen sections. The supplies tend to be on the expensive side but it doesn't look like the cost has gone up too much in the last ten years. I was pleased enough with both the product and the customer service that I just recently purchased both the paraffin and the frozen apparatus for my lab in Nebraska. The only problem I ever had, after initially learning the technique, was the matrix of the 'tape' or the slide showed up when I did fluorescent staining. It had a honeycomb appearance under the FITC wavelength that we were viewing and it was hard to see the positive staining. When I presented this problem to the R&D people at Instrumedics, they developed a mounting media that did a good job in alleviating it. I work with whole mouse embryos so I had to customize the standard procedure a bit. I am willing to share more of the intricacies of my Instrumedics experience with you and/or the scientist in personal correspondence if you would like specifics. Anita One of the scientist I work with is looking into purchasing a cryojane. Does anyone have any experiance with this piece of equipment. Also the gentle jane for snap frozens. Steve --------------------------------- Start your day with Yahoo! - make it your home page _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rachael_Emerson <@t> URMC.Rochester.edu Tue Aug 9 12:03:47 2005 From: Rachael_Emerson <@t> URMC.Rochester.edu (Emerson, Rachael) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] activated notch/trilogy Message-ID: Hello. Has anyone worked with Cleaved Activated Notch antibody or the unmasking solution, Trilogy, from Cell Marque? I am having problems with background and could use some assistance. Thank you Rachael Rachael L. Emerson Center for Pediatric Biomedical Research University of Rochester Medical Center 575 Elmwood Avenue MRBX 1.11301 Rochester, NY 14642 Tel (585) 275-5073 Fax (585) 276-0232 From ajohnson <@t> aipathology.com Tue Aug 9 12:11:20 2005 From: ajohnson <@t> aipathology.com (Amy Johnson) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Reagent labels Message-ID: <016A64931E1ED511B12C0002B30260805233A9@SERV001> We are busy updating things around the lab according to the new CAP checklist. One of the things our supervisor wants us to do now is to label our frozen section line with date made, and date of expiration. The techs feel that this is too much information to put on a lid and were wondering if a log book could be used instead. If an expiration date is used is it necessary for a preparation date as well. Thanks Amy Johnson From shahn14 <@t> jhmi.edu Tue Aug 9 12:20:00 2005 From: shahn14 <@t> jhmi.edu (Sven Hahn) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] 3-Nitrotyrosine Message-ID: Hello. Has anyone worked with 3-Nitrotyrosine-antibody in humans and paraffin sections. I am having even problems to stain the pos. control and could use some assistance. Thank you Sven Hahn JHU Baltimore From vazquezr <@t> ohsu.edu Tue Aug 9 14:15:34 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Reagent labels Message-ID: Amy, Why on the stain line itself? An expiration date and date made on the bottle should be sufficient. Or take 2 strips of tape and place on lid/down the side, then write the dates on each one. That's what I do when I use a solution continuously. Robyn OHSU >>> "Amy Johnson" 8/9/2005 10:11 AM >>> We are busy updating things around the lab according to the new CAP checklist. One of the things our supervisor wants us to do now is to label our frozen section line with date made, and date of expiration. The techs feel that this is too much information to put on a lid and were wondering if a log book could be used instead. If an expiration date is used is it necessary for a preparation date as well. Thanks Amy Johnson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Tue Aug 9 14:53:04 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Slide cabinets with locks Message-ID: Does anyone know of a source for slide cabinets that can be locked? Thank you! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From JMacDonald <@t> mtsac.edu Tue Aug 9 16:13:32 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Job Posting Northern California In-Reply-To: Message-ID: ? Histology Technician ? Full-time, day shift QUALIFICATIONS: HT or HTL ASCP certification or eligibility preferred. Previous Histology experience preferred. JOB DUTIES: Technical and clerical duties in the Histology and Cytology sections of the lab, including, but not limited to, accessioning, embedding, sectioning, staining, coverslipping and immunohistochemical staining of Pathology specimens. EQUIPMENT: ? Ventana Benchmark IHC Stainer. ? Leica 2025 Microtomes. ? Thermo Shandon Excelsior Tissue Processor. ? Thermo Shandon Embedding Center ? TriPath SurePath Liquid Pap Smear Stainer. SKILLS REQUIRED: ? Ability to read and interpret documents such as safety rules, operating and maintenance instructions, and procedure manuals. ? Ability to write complex reports and correspondence. ? Ability to perform complex mathematical calculations. ? Ability to apply common-sense understanding to carry out instructions furnished in written, oral or diagram form. ? Ability to deal with problems involving many concrete variables in standardized situations. Humboldt County, the ideal setting to launch your career. Think of fresh air, the Pacific coastline and ancient Redwood forests. Then add competitive compensation and benefits, quality schools, plus employment information for your spouse or partner. The change is in your favor at St. Joseph Health System, where professional fulfillment and quality of life come hand-in-hand at St. Joseph Hospital, Eureka. We have the following outstanding opportunity for someone with the desire to be a part of a healthcare team, yet can also think and act independently. Please send your resume to: Human Resources, St. Joseph Hospital, 2700 Dolbeer, Eureka, CA 95501. Email: Meri.Scolari@stjoe.org Phone: (707) 269-4271. Contact the Pathology Supervisor @ 707-441-4420 ext.6621 or email: CPETTY@SJE.STJOE.ORG for more information. From jqb7 <@t> cdc.gov Tue Aug 9 20:04:55 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Slide cabinets with locks Message-ID: Sakura has one. I will try to send the attachment but if it the picture doesn't display just go to www.sakuraus.com and open their electronic catalog and look for Lab Aid Ultra. They have one for cassettes as well. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Richard Cartun Sent: Tue 8/9/2005 3:53 PM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Slide cabinets with locks Does anyone know of a source for slide cabinets that can be locked? Thank you! Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -------------- next part -------------- Sakura - View Items
Item No. Description Alt. Item No. Quantity Price
3991 LAB AID® ULTRA™ CABINET FOR SLIDES 1 EACH $3,690.00

LAB AID® ULTRA™ CABINET FOR SLIDES

The Lab Aid® Ultra™ Slide and Block Storage System is a modernized system offering large capacity storage for slides and blocks with integrated security features. A sturdy steel construction ensures long term use and a drawer lock prevents tampering, guaranteeing specimen safety. Each drawer front has a plastic covered labeling tab assuring positive identification. Inside, convenient removable slide and block trays accommodate up to 82,000 microscope slides or 21,000 blocks. Coding strips can be used to separate slides or blocks by numbers. Filing cabinets can be bolted together and stacked two high for maximum storage capacity.

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From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Aug 10 05:54:58 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Orcein for copper-binding protein/Hep B Message-ID: Hello Histonetters. Has anyone got a really good orcein method for copper-binding protein/Hep B, and is there a recommended brand or vendor of the dye? Would you use synthetic or natural (if you can still get it)? Cheers Jacqui Malam Lancaster Infirmary N England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From BoozerKA <@t> pa1.ah.org Wed Aug 10 06:58:02 2005 From: BoozerKA <@t> pa1.ah.org (Kathleen Boozer) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Green Metal Slide Trays Message-ID: Adventist Medical Center in Portland, Oregon has 120 green metal slide trays with 10 bases. Went from 20+ years down to 10 storage. Make offer. Kathleen Boozer 503 251-6266 ex. 7582 From lilliangarrett <@t> gmail.com Wed Aug 10 07:24:21 2005 From: lilliangarrett <@t> gmail.com (Lillian Garrett) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Re: Still desperately trying to optimise Vasopressin receptor stain Message-ID: <81b03aa30508100524741156cb@mail.gmail.com> Hi everybody, I'm just wondering whether anyone has had success using the vasopressin V1b receptor antibody from Alpha Diagnostics International in brain tissue... I am having trouble optimising this stain and get faint to no signal...... Could anyone suggest possible ways of getting the most from this staining..... The brain tissue is fixed in 4% paraformaldehyde.... I want to stain cryosectioned material using DAB as the cromogen.... So far I have increased the primary antibody incubation time to 48 hours and have heated the sections to no avail..... Any help with this problem would be greatly appreciated... Thanks, Lillian From John.Auld <@t> whnt.nhs.uk Wed Aug 10 08:42:17 2005 From: John.Auld <@t> whnt.nhs.uk (John Auld) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Re: Problems with certification Message-ID: Hi Meryl I'm a bit behind with Histonet at the moment so excuse the delay. Our colleagues in the USE will no doubt offer advice on how to overcome your situation, but why do you want to go somewhere were they don't appear to want you? There is a shortage of histotechs here in the UK so you should be able to get a job suitable to your experience and abilities within a couple of hundred miles rather than several thousand. This makes it much easier to go hoem if things don't turn out as you'd hoped. Good luck in your search John John Auld MSc CSci FIBMS BMS 4 Dept of Histopathology and Clinical Cytology Arrowe Park Hospital Arrowe Park Road Upton Wirral Internal extn 2560 External Tel 0151 604 7025 Message: 8 Date: Thu, 04 Aug 2005 08:31:57 +0100 From: "Meryl Roberts" Subject: [Histonet] Problems with certification To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed Hi there, I am a histologist in the UK with nearly 7 years experience, but I wish to move to the USA. However I'm having terrible problems finding work as it seems it's virtually impossible for me to become certified in the USA as none of my work experience is in an American lab- I've spoken to the ASCP and the NCA and they both say that I'm not elligeable to apply, and I'm currently waiting to hear back from the AMT (fingers crossed!).....is there any way I can become certified or should I just give up? I'm starting to get a bit despondant. sincerely, Meryl Roberts. From vazquezr <@t> ohsu.edu Wed Aug 10 09:04:14 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Green Metal Slide Trays Message-ID: Kathleen, Are they the Tissue-Tek type, dark green? Robyn OHSU 503-494-4658 >>> "Kathleen Boozer" 8/10/2005 4:58 AM >>> Adventist Medical Center in Portland, Oregon has 120 green metal slide trays with 10 bases. Went from 20+ years down to 10 storage. Make offer. Kathleen Boozer 503 251-6266 ex. 7582 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pex0220 <@t> yahoo.com.cn Wed Aug 10 09:51:29 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] I would like to talk about immunofluorescence in bone or cartilage paraffin sections! Message-ID: <20050810145129.91268.qmail@web15510.mail.cnb.yahoo.com> Hello, all. I have done immunofluorescence in bone or cartilage paraffin sections for several months. I use normal immunofluorescence protocol for staining, but I have tried different methods for antigen retrieval, finally I think that 0.1% trypsin (15min or 20min) for antigen retrieval in my experiments is a little bit good. In addition, I have already used different antibodies for immunosatining. One of them is working well, positive expressions can be found in cartilage, osteoblasts and osteocytes, but others express in cartilage strongly, however they are very weak in bone( but they express stronger in primary osteoblasts).I do not know the reasons! Any suggestions will be useful for me! Thank you! Guofeng --------------------------------- DO YOU YAHOO!? ÑÅ»¢Ãâ·ÑGÓÊÏ䣭ÖйúµÚÒ»¾øÎÞÀ¬»øÓʼþɧÈų¬´óÓÊÏä From Michele_Marggi <@t> ssmhc.com Wed Aug 10 09:58:07 2005 From: Michele_Marggi <@t> ssmhc.com (Michele_Marggi@ssmhc.com) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] HT position available Message-ID: There is a part time HT position available at St. Marys Hospital Medical Center in Madison, WI. Below is a brief summary of the position. If you are interested- or know someone that may be....please contact me... Opportunities are available for an individual to practice and enhance their histology skills in the preparation of tissues for microscopic examination. Some of the duties include: embedding, cutting, staining and coverslipping of routine surgical specimens, perform special and immunoperoxidase stains which may include the utilization of automated equipment. Prepares stock solutions and special stains. Prepares frozen sections and assists in collection of Fine Needle Aspirate specimens. Require strong interpersonal, communication, and problem solving skills. HT or MLT with ASCP certification or eligibility required. Our Laboratory handles over 38,000 surgical cases a year utilizing state of the art technology. The successful candidate should be able to process a high volume of cases in a short period of time with an emphasis on quality and accuracy. This position is 60 hours in two weeks with flexible 6 hour shifts scheduled any time between 4:30 a.m. and 3:00 p.m.; Monday through Friday. For more information, please go to: http://www.stmarysmadison.com (click on employment) Or contact me directly with questions! Michele Marggi Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email 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 email and destroy all copies of the original message. From jkiernan <@t> uwo.ca Wed Aug 10 11:02:40 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Orcein for copper-binding protein/Hep B References: Message-ID: <42FA2520.DB87734B@uwo.ca> Orcein has always been a synthetic dye. The starting material in manufacture is the phenolic compound orcinol (3,5-dihydroxytoluene). Orcinol was originally obtained by alkaline hydrolysis and decarboxylation of a more complex compound that occurs in certain lichens, including one called orchil. Oxidaton of orcinol in the presence of ammonia yields orcein, which is a mixture of 14 oxazine dyes. Related dye mixtures are lacmoid and litmus, for which the starting material is resorcinol (1,3-dihydroxybenzene). The medieval method for making orcein involved boiling the lichen in alkaline urine, a procedure that combined hydrolysis of the precursor with oxidation in the presence of a source of ammonia. Although orcein is sometimes designated "synthetic" on the label, this has never been a "natural" dye in the sense of being responsible for the colour of an animal or plant. The lichen (orchil) was simply a source of a precursor of orcinol (which is not coloured) in the years before coal tar became the major source of aromatic compounds. Orcein is one of the dyes regularly tested by the Biological Stain Commission, and the tests include methods for elastin and hepatitis A antigen. See Penney, Powers, Frank & Churukian 2002 Biotech. Histochem. 77:237-275 for the staining methods used to do the testing. Any bottle of orcein with a "Certified" label will therefore be suitable. The December 2003 issue of Biotechnic & Histochemistry (Vol 78 No. 6) was a special issue with six papers on orcein and related dyes. One of these, by Tony Henwood (pp. 303-308), addresses batch variation and staining by aged solutions of orcein, with coloured photos showing the varied results. -- ------------------------------- 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 _______________________________ Malam Jacqueline wrote: > > Hello Histonetters. > Has anyone got a really good orcein method for copper-binding protein/Hep B, > and is there a recommended brand or vendor of the dye? Would you use > synthetic or natural (if you can still get it)? > Cheers > Jacqui Malam > Lancaster Infirmary > N England > > DISCLAIMER: This e-mail is confidential and privileged. If you are not the > intended recipient please accept our apologies; please do not disclose, copy > or distribute information in this e-mail or take any action in reliance on > its contents: to do so is strictly prohibited and may be unlawful. Please > inform postmaster@rli.mbht.nhs.uk that this message has gone astray before > deleting it. Comments or opinions expressed in this email are those of > their respective contributors only. The views expressed do not represent the > views of the Trust, its management or employees. Morecambe Bay Hospitals NHS > Trust is not responsible and disclaims any and all liability for the content > of comments written within.Thank you for your co-operation. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From beckyjo <@t> email.unc.edu Wed Aug 10 11:06:18 2005 From: beckyjo <@t> email.unc.edu (Rebecca Jo) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Sections falling off during in situ Message-ID: <20050810120618.kqrnupw7oc0kgoo0@webmail1.isis.unc.edu> Hello all- I'm doing an in situ hybridization with adult frog eye sections. I cut the sections on a cryostat about 20 microns thick. I've been having problems with my sections falling off after treatment with proteinase K. My protocol calls for 10ug/ml of proteinase K; however, I have played around with different concentrations, even going down to 1ug/ml. It seems no matter what I do, my sections tend to fall off. Please, what am I doing wrong? Also if it helps, before treatment with proteinase K, I dry the sections at 50C for 10 minutes then and treat with 4% PFA 20 minutes. Thanks all. -- Rebecca E. Jo UNC-School of Medicine Department of Cell and Developmental Biology (919)843-9648 CB# 7090 From dlcowie <@t> prodigy.net Wed Aug 10 11:10:42 2005 From: dlcowie <@t> prodigy.net (Dawn Cowie) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] histology position available Message-ID: <20050810161042.73789.qmail@web81001.mail.yahoo.com> hello all histonetters, we have a position open for a ascp certified HT. Must either be Florida licensed or willing to obtain license. This is a full time 40 hrs per week position with generous PTO, medical, dental, short and long term disability and 401K and profit sharing. Monday thru Friday and half a day Saturday about every 4 weeks. Duties include, embedding, microtomy, special stains, and IHC. We are located in the panhandle area of northwest Florida. If anyone is interested or wants more details, please call me at 850-416-7251 or e-mail me at dlcowie@prodigy.net Thanks, Dawn Cowie, HT. Histology Supervisor Pensacola Pathologists, PA From T.J.A.vanEijl <@t> pharm.uu.nl Wed Aug 10 11:23:51 2005 From: T.J.A.vanEijl <@t> pharm.uu.nl (Eijl, drs. T.J.A. van) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Apoptosis detection in plastic embedded tissue Message-ID: Dear histonetters, We have been trying to stain apoptotic cells in Carnoy fixed, BMA/MMA embedded mouse lungs using the chromogenic DEAD-END Kit by Promega. However, this kit has been designed for formalin fixed, parafin embedded tissue. To cut a long story short, we did not get it to work properly. Now I am enquiring if anybody has knowledge of an apoptosis detection method that can be used on plastic embedded tissues as described above. Thanks in advance, Sven van Eijl Ph.D. student Dept. of Pharmacology & Pathophysiology UIPS The Netherlands From ree3 <@t> leicester.ac.uk Wed Aug 10 11:29:44 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Immunostaining of peripheral blood films Message-ID: I have no experience of the above, any tips gratefully received. Thanks Richard Edwards MRC TOXICOLOGY UNIT LEICESTER. U.K.... From cbass <@t> bidmc.harvard.edu Wed Aug 10 12:49:12 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] stereotactic frame Message-ID: <53721EC0-4A3A-40B2-B7A1-4EB976C6FFF2@bidmc.harvard.edu> Hello, Does anyone have advice for stereotactic frames? We need one in the lab and I would like opinions about reputable companies. I have never purchased a frame before. It's a major purchase for the lab so I need to find something that is rock solid. I am looking for something that will work with both mice and rats for standard brain injections. Any advice would be appreciated. Thanks, Caroline Bass From BoozerKA <@t> pa1.ah.org Wed Aug 10 12:50:25 2005 From: BoozerKA <@t> pa1.ah.org (Kathleen Boozer) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Green Metal Slide Trays Message-ID: 5" high x 15 ?" wide x18 3/4" long Paragon - 6 2-side slide columns per tray. >>> "Robyn Vazquez" 08/10/05 7:04 AM >>> Kathleen, Are they the Tissue-Tek type, dark green? Robyn OHSU 503-494-4658 >>> "Kathleen Boozer" 8/10/2005 4:58 AM >>> Adventist Medical Center in Portland, Oregon has 120 green metal slide trays with 10 bases. Went from 20+ years down to 10 storage. Make offer. Kathleen Boozer 503 251-6266 ex. 7582 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Wed Aug 10 14:18:50 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] RE: Cleaved notch antibody/Trilogy Message-ID: Rachael, are you really using undiluted biotinylated goat anti-rabbit and undiluted streptavidin-HRP from Zymed? If so, that's your problem. Those are concentrated reagents and are intended to be diluted before use (from the data sheets I looked at, the concentration of the secondary antibody is 0.75 mg/ml; the streptavidin is recommended to be diluted at 1:150 - 1:750). I'd start there and see if this helps. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org -----Original Message----- From: Emerson, Rachael [mailto:Rachael_Emerson@URMC.Rochester.edu] Sent: Wednesday, August 10, 2005 1:43 PM To: Johnson, Teri Subject: RE: Cleaved notch antibody/Trilogy Hi Teri. Here is the protocol that I have been using. It was derived from a lab looking at the facial features of embryonic mice. <> Also note that I did some trials in the following manner and still saw a ton of background: Deparafinize & rehydrate to PBS Trilogy vs Citrate Buffer vs. no antigen retrieval Rinse in water, into PBS Blocked with 3% H2O2 in MeOH Rinsed with PBS Incubated with Streptavidin-HRP for 30' Rinsed in PBS Developed in DAB Any thoughts would be appreciated! Thanks!!!! Rachael Emerson Rachael L. Emerson Center for Pediatric Biomedical Research University of Rochester Medical Center 575 Elmwood Avenue MRBX 1.11301 Rochester, NY 14642 Tel (585) 275-5073 Fax (585) 276-0232 From GDawson <@t> dynacaremilwaukee.com Wed Aug 10 14:17:26 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] BK polyoma virus Message-ID: All, Besides Chemicon, is there another vendor that carries BK virus for FFPE tissues? Thanx In Advance for any responses, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI From sa.drew <@t> hosp.wisc.edu Wed Aug 10 14:32:53 2005 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] BK polyoma virus Message-ID: We are using Calbiochem's PAb416 monoclonal...BK/SV40 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dawson, Glen Sent: Wednesday, August 10, 2005 2:17 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] BK polyoma virus All, Besides Chemicon, is there another vendor that carries BK virus for FFPE tissues? Thanx In Advance for any responses, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Wed Aug 10 14:41:17 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] BK polyoma virus In-Reply-To: Message-ID: Access Biomedical carries this antibody. Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA > All, > > Besides Chemicon, is there another vendor that carries BK virus for FFPE > tissues? > > Thanx In Advance for any responses, > > Glen Dawson BS, HT & QIHC (ASCP) > IHC Manager > Milwaukee, WI > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nancy.troiano <@t> yale.edu Wed Aug 10 14:45:30 2005 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] apoptosis detection in MMA embedded tissue Message-ID: <5.2.1.1.2.20050810154334.00bdfdf0@email.med.yale.edu> Years ago we used the Klenow FragEL DNA fragmentation detection kit, Catalog #QIA21 on MMA embedded mouse bones to detect apoptotic cells with success. The vendor is Oncogene Research Products, Boston ,Massachusetts, USA. From tessajmurray <@t> hotmail.com Wed Aug 10 15:50:00 2005 From: tessajmurray <@t> hotmail.com (Tessa Murray) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Cryostat sectioning of mammary gland Message-ID: I'm looking for some advice on cryosections. We are trying to collect frozen sections of mammary gland tissue and are experiencing problems - we do a lot of paraffin work here so the cryostat technique is quite new to us. We find that the knife cuts the OCT fine, but when it hits tissue it does not cut so we get sections with tissue shaped holes in them. We've tried changing the block and knife temp (we have tried altering them between -30 to -35C) but nothing seems to help. Any advice appreciated... tess From jryan <@t> sleh.com Wed Aug 10 17:13:36 2005 From: jryan <@t> sleh.com (John Ryan) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Position available in Houston Message-ID: Histology technician or technologists position available at St. Luke's Episcopal Health System in Houston, Texas. This is a day position in a high volume busy laboratory with the bulk of the H&E stains done during the night. If interested please contact John Ryan @ 832-355-2643 or email at jryan@sleh.com John P Ryan, MBA, HT(ASCP)HTL Assistant Administrative Director Pathology St. Luke's Episcopal Hospital 832-355-2643 832-355-4232 fax jryan@sleh.com "CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient (s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message." +++++CONFIDENTIALITY NOTICE+++++ The information in this e-mail may be confidential and/or privileged. If you are not the intended recipient or an authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments, if any, or the information contained herein is prohibited. If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail from your computer system. Thank you. From vazquezr <@t> ohsu.edu Wed Aug 10 17:24:38 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Cryostat sectioning of mammary gland Message-ID: Tessa, have you tried tightening the blade holder? robyn OHSu >>> "Tessa Murray" 8/10/2005 1:50 PM >>> I'm looking for some advice on cryosections. We are trying to collect frozen sections of mammary gland tissue and are experiencing problems - we do a lot of paraffin work here so the cryostat technique is quite new to us. We find that the knife cuts the OCT fine, but when it hits tissue it does not cut so we get sections with tissue shaped holes in them. We've tried changing the block and knife temp (we have tried altering them between -30 to -35C) but nothing seems to help. Any advice appreciated... tess _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Wed Aug 10 17:47:04 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Cryostat sectioning of mammary gland Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175A3@lsexch.lsmaster.lifespan.org> The problem with sectioning mammary gland is, in a word, FAT. Fat doesn't freeze at -20 degrees. It also doesn't freeze very well at -35 degrees. A mammary gland where fibrosis or tumor has replaced a lot of the fat cuts quite well. But typical mammary gland does not. First, you will need to get the block even colder, by using a freon spray. This can bring the block temperature down to about -50 degrees. If you are using an anti-roll plate, spray that first, generously, and wipe off any excess refrigerant. Then spray the block generously for a few seconds, then IMMEDIATELY cut your section, before the block can warm at all. Also, it may not be possible to cut 4 or 5 micron sections. A 10-15 micron setting will give you a much better chance of getting a section of fatty tissue. Cutting the sections faster also frequently helps with tissue of this type. Freeze the block with the spray, then turn the handle quickly, so that you "chop" the section off the block (so to speak) rather than "slice" it off. Finally, it usually isn't the fat that you are interested in, when sectioning this kind of tissue. You may find that even though there are holes where some of the larger fat masses were, the stroma is still intact in the section. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tessa > Murray > Sent: Wednesday, August 10, 2005 1:50 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Cryostat sectioning of mammary gland > > > I'm looking for some advice on cryosections. We are trying to collect > frozen sections of mammary gland tissue and are experiencing problems > - we do a lot of paraffin work here so the cryostat technique is quite > new to us. We find that the knife cuts the OCT fine, but when it hits > tissue it does not cut so we get sections with tissue shaped holes in > them. We've tried changing the block and knife temp (we have tried > altering them between -30 to -35C) but nothing seems to help. Any > advice appreciated... > > tess > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From petepath <@t> yahoo.com Wed Aug 10 18:10:52 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Cryostat sectioning of mammary gland Message-ID: <20050810231052.7941.qmail@web30412.mail.mud.yahoo.com> Hi Tessa, I have a comprehensive tutorial on frozen section technique on my web site which I think you will find useful as a starting point. http://pathologyinnovations.com/frozen_section_technique.htm I address cutting fatty tissues from the standpoint of a surgical pathologist which with good technique will get you very acceptable sections. If you need absolute perfection with very fatty tissues I think your best bet is Instrumedics tape sytem. I have no experience with it but I believe it is suppossed to work with fatty tissue. Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From kccatunda <@t> terra.com.br Wed Aug 10 19:57:50 2005 From: kccatunda <@t> terra.com.br (Katia Cristina Catunda) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Help on Workload - CAP References: <20050810145129.91268.qmail@web15510.mail.cnb.yahoo.com> Message-ID: <000e01c59e0f$b2ddb720$2c65fea9@privatexx> Searching for a workload parameter to analyse if we do have many or less histotechnicians as necessary for our routine I found one "Q & A" answer in CAP (cited in the end of the email). Our slide-average per histotechnician is about 40 slides per day and I don't see how I can reduce our staff to get the 50 slides suggested by CAP. We do have at the same time 2 technicians on embedding (starting at 6 am), 4 techinicians on microtomy (starting on 7 am) and 2 technicians on staining and mounting (starting on 7 am). We don't have any automatization on staining neither mounting. We release our firsts slides at 8h20 and they are continuously liberated until the end of the routine (about 12 pm). The technicians who does embedding does special stains and all the processor's battery-change. The technicians that stain and mount the slides does the archivation of the cassettes. And at 12pm we start our microwave routine with the same technicians that processed the morning-routine. Can anyone give me some tips... are we so away from the average workload of the other labs? How can I evalue it precisely? Pleeeeassseee... heeeelllpp!!! lol Thanks you all, Katia __________________________________________________________________________ Q. How many cassettes should be processed by a histotechnologist during an eight-hour shift? A. No established comprehensive standard addresses histology workload. Previously published CAP workload guidelines for histopathology, based on data from the Laboratory Management Index Program, say "each well-trained HT/ HTL can be expected to produce approximately 3,000 slides per quarter, or 12,000 slides per year. Included in these totals are 2,500 H&E slides and 500 common special stains, or 10,000 H&E slides and 2,000 common special stains. If only rare special stains are requested, more set-up time is required. Twelve thousand slides per year is equivalent to 50 slides per day. These are average numbers for a lab where much of the work is not automated." In practice, a uniform standard across laboratories may be an unrealistic goal because many factors influence the number of blocks a histotechnician/technologist can cut in a given period. These include: a.. The experience level of the technician. A new employee or student would be expected to cut at a slower rate. b.. The case complexity. Biopsies, which require multiple levels and careful trimming, require considerably more time than routine cases (for example, uterus). c.. The number of interruptions. Smaller laboratories, in which the cutting technician may be answering the phone or receiving special stain or recut requests, or both, will have lower productivity. The most useful standard for employees in a given laboratory is set by the supervisor or senior technologists, or both, based on past productivity levels. To address productivity in a more global sense, it is necessary to assign work units to each of the varied tasks in histology, including loading and maintaining processors, embedding, cutting, routine staining, special stains, and immunohistochemistry. It is then possible to benchmark units worked per hour. As with routine cutting, however, the assignment of work unit values to a given task can only be done realistically by the histology supervisor and pathologist at a given site, taking into account economies of scale and levels of automation. Richard W. Brown, MD Medical Director, Core Histology Laboratory Memorial Hermann Healthcare System Houston Member, CAP Surgical Pathology Committee With members of the CAP/National Society for Histology Committee: Freida L. Carson, PhD, HT(ASCP) Lena T. Spencer, MA, HT(ASCP)HTL, QIHC Vincent Della Speranza, MMS, HT(ASCP)HTL Sue E. Lewis, HTL(ASCP) From lpwenk <@t> sbcglobal.net Wed Aug 10 20:32:59 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Orcein for copper-binding protein/Hep B In-Reply-To: Message-ID: Are you looking for copper or for the hepatitis B? If I remember correctly, orcein stains the hepatitis B inclusion body (the ground glass appearance on an H&E)and it also stains elastin. Other stains for the hepatitis B were the aldehyde fuchsin and the Victoria blue. I think most labs do an IHC for hepatitis, as you can differentiate Hepatitis B core from Hepatitis B surface antigens, as well as differentiate it from Hepatitis A, C or D. None of these stained for copper, such as in Wilson's disease. For that, we use rhodanine. I believe it did not stain the actual copper, but rather the protein to which copper was bound. Let me know what procedures you need. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak MI 48073 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malam Jacqueline Sent: Wednesday, August 10, 2005 6:55 AM To: Histonet submissions (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Orcein for copper-binding protein/Hep B Hello Histonetters. Has anyone got a really good orcein method for copper-binding protein/Hep B, and is there a recommended brand or vendor of the dye? Would you use synthetic or natural (if you can still get it)? Cheers Jacqui Malam Lancaster Infirmary N England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CrochiereSteve <@t> aol.com Wed Aug 10 21:47:44 2005 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] H. Pylori IHC Message-ID: <9f.64cff233.302c1650@aol.com> I have been getting a lot of background in the mucosa of the Gastric biopsies, especially in cases that show increased inflammation and many eosinophils. I have increased the incubation time in peroxide blocker and protein blocker and decreased the time in the primary and secondary Ab, only to find it getting worse instead of better. I am using Biocare's antibody and detection system on their instrument. Any suggestions to eliminate this nonspecific staining? From andromeda_tm <@t> libero.it Thu Aug 11 06:03:22 2005 From: andromeda_tm <@t> libero.it (Massimo) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Histoclear, instead of Xilene... Message-ID: Torino 11 August 2005 (ITALY) Dear Histoneters, In the operation of wax inclusion I have substituted the Xilene with a commercial (Brunel Microscopes Ltd.) product: "Histoclear". I wonder if the time of treatment would be the same as with Xilene or longer. I am talking of a specimen volume like one or half cubic centimetre (for instance, now I am processing a frog tadpole) I would appreciate some suggestions. Thank you. With my Best Regards, Massimo From BZIMMERM <@t> mail.mcg.edu Thu Aug 11 07:14:34 2005 From: BZIMMERM <@t> mail.mcg.edu (Billie Zimmerman) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Help--Need BK postive control tissue Message-ID: Does anyone know where I could find a positive control for BK?? Thanks, Billie Zimmerman MT(ASCP) QIHC From JMahoney <@t> alegent.org Thu Aug 11 08:41:39 2005 From: JMahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Help on Workload - CAP Message-ID: Hello Katia and All, I have been amazed at what the HT's at my institution have been able to accomplish over the past year. We have implemented LEAN in our laboratory. I'm presenting a workshop at the National meeting in Ft. Lauderdale next month on how we accomplished reducing our TAT, space, cost etc. The main impact has come form implementing "single piece flow" of specimens. It is amazing the difference it makes to eliminate "waste". This "waste" includes, excess walking, blocks/slides just sitting with nothing being done with them, techs waiting for work and getting rid of unnecessary steps in the process. These are just a few thoughts on how to help with your issue. Good Luck. Jan, Omaha >>> "Katia Cristina Catunda" 08/10/2005 7:57:50 PM >>> Searching for a workload parameter to analyse if we do have many or less histotechnicians as necessary for our routine I found one "Q & A" answer in CAP (cited in the end of the email). Our slide-average per histotechnician is about 40 slides per day and I don't see how I can reduce our staff to get the 50 slides suggested by CAP. We do have at the same time 2 technicians on embedding (starting at 6 am), 4 techinicians on microtomy (starting on 7 am) and 2 technicians on staining and mounting (starting on 7 am). We don't have any automatization on staining neither mounting. We release our firsts slides at 8h20 and they are continuously liberated until the end of the routine (about 12 pm). The technicians who does embedding does special stains and all the processor's battery-change. The technicians that stain and mount the slides does the archivation of the cassettes. And at 12pm we start our microwave routine with the same technicians that processed the morning-routine. Can anyone give me some tips... are we so away from the average workload of the other labs? How can I evalue it precisely? Pleeeeassseee... heeeelllpp!!! lol Thanks you all, Katia __________________________________________________________________________ Q. How many cassettes should be processed by a histotechnologist during an eight-hour shift? A. No established comprehensive standard addresses histology workload. Previously published CAP workload guidelines for histopathology, based on data from the Laboratory Management Index Program, say "each well-trained HT/ HTL can be expected to produce approximately 3,000 slides per quarter, or 12,000 slides per year. Included in these totals are 2,500 H&E slides and 500 common special stains, or 10,000 H&E slides and 2,000 common special stains. If only rare special stains are requested, more set-up time is required. Twelve thousand slides per year is equivalent to 50 slides per day. These are average numbers for a lab where much of the work is not automated." In practice, a uniform standard across laboratories may be an unrealistic goal because many factors influence the number of blocks a histotechnician/technologist can cut in a given period. These include: a.. The experience level of the technician. A new employee or student would be expected to cut at a slower rate. b.. The case complexity. Biopsies, which require multiple levels and careful trimming, require considerably more time than routine cases (for example, uterus). c.. The number of interruptions. Smaller laboratories, in which the cutting technician may be answering the phone or receiving special stain or recut requests, or both, will have lower productivity. The most useful standard for employees in a given laboratory is set by the supervisor or senior technologists, or both, based on past productivity levels. To address productivity in a more global sense, it is necessary to assign work units to each of the varied tasks in histology, including loading and maintaining processors, embedding, cutting, routine staining, special stains, and immunohistochemistry. It is then possible to benchmark units worked per hour. As with routine cutting, however, the assignment of work unit values to a given task can only be done realistically by the histology supervisor and pathologist at a given site, taking into account economies of scale and levels of automation. Richard W. Brown, MD Medical Director, Core Histology Laboratory Memorial Hermann Healthcare System Houston Member, CAP Surgical Pathology Committee With members of the CAP/National Society for Histology Committee: Freida L. Carson, PhD, HT(ASCP) Lena T. Spencer, MA, HT(ASCP)HTL, QIHC Vincent Della Speranza, MMS, HT(ASCP)HTL Sue E. Lewis, HTL(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjchtascp <@t> yahoo.com Thu Aug 11 09:12:19 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Orcien/Iron Hematoxylin elastic stain Message-ID: <20050811141219.21726.qmail@web90209.mail.scd.yahoo.com> Does anyone have the formulation for the orcein plus iron hematoxylin elastic stain. I'm unable to find my copy. Steve --------------------------------- Start your day with Yahoo! - make it your home page From abright <@t> brightinstruments.com Thu Aug 11 09:21:22 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Cryostat sectioning of mammary gland Message-ID: Dear Tess, Are you sure the cryochamber is below -30?C or just the specimen is at that temperature?? As your problem indicates that the knife & anti-roll plate are too warm, if the specimen is at -30?C. With the correct temperatures, mammary gland should be easy to section. 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: Tessa Murray [mailto:tessajmurray@hotmail.com] Sent: 10 August 2005 21:50 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cryostat sectioning of mammary gland I'm looking for some advice on cryosections. We are trying to collect frozen sections of mammary gland tissue and are experiencing problems - we do a lot of paraffin work here so the cryostat technique is quite new to us. We find that the knife cuts the OCT fine, but when it hits tissue it does not cut so we get sections with tissue shaped holes in them. We've tried changing the block and knife temp (we have tried altering them between -30 to -35C) but nothing seems to help. Any advice appreciated... tess _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MLafrini <@t> csmlab.com Thu Aug 11 09:20:33 2005 From: MLafrini <@t> csmlab.com (Michael LaFriniere) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Help on Workload - CAP Message-ID: Katia, As chairperson for the NSH Productivity Task force, The December 2004 Issue of "The Journal of Histotechnology" "Special Report" may be of benefit to you. We performed a nation wide study over a year period on the issues you addressed concerning productivity in the histology laboratory. You can probably obtain a copy of the report if you do not have this issue by calling the NSH office. Sincerely, Michael R. LaFriniere Executive Director Cytology Services of Maryland (CSM) 301-206-2555 ext 27 301-206-2595 fax MLafrini@csmlab.com >>> "Katia Cristina Catunda" 08/10/05 8:57 PM >>> Searching for a workload parameter to analyse if we do have many or less histotechnicians as necessary for our routine I found one "Q & A" answer in CAP (cited in the end of the email). Our slide-average per histotechnician is about 40 slides per day and I don't see how I can reduce our staff to get the 50 slides suggested by CAP. We do have at the same time 2 technicians on embedding (starting at 6 am), 4 techinicians on microtomy (starting on 7 am) and 2 technicians on staining and mounting (starting on 7 am). We don't have any automatization on staining neither mounting. We release our firsts slides at 8h20 and they are continuously liberated until the end of the routine (about 12 pm). The technicians who does embedding does special stains and all the processor's battery-change. The technicians that stain and mount the slides does the archivation of the cassettes. And at 12pm we start our microwave routine with the same technicians that processed the morning-routine. Can anyone give me some tips... are we so away from the average workload of the other labs? How can I evalue it precisely? Pleeeeassseee... heeeelllpp!!! lol Thanks you all, Katia __________________________________________________________________________ Q. How many cassettes should be processed by a histotechnologist during an eight-hour shift? A. No established comprehensive standard addresses histology workload. Previously published CAP workload guidelines for histopathology, based on data from the Laboratory Management Index Program, say "each well-trained HT/ HTL can be expected to produce approximately 3,000 slides per quarter, or 12,000 slides per year. Included in these totals are 2,500 H&E slides and 500 common special stains, or 10,000 H&E slides and 2,000 common special stains. If only rare special stains are requested, more set-up time is required. Twelve thousand slides per year is equivalent to 50 slides per day. These are average numbers for a lab where much of the work is not automated." In practice, a uniform standard across laboratories may be an unrealistic goal because many factors influence the number of blocks a histotechnician/technologist can cut in a given period. These include: a.. The experience level of the technician. A new employee or student would be expected to cut at a slower rate. b.. The case complexity. Biopsies, which require multiple levels and careful trimming, require considerably more time than routine cases (for example, uterus). c.. The number of interruptions. Smaller laboratories, in which the cutting technician may be answering the phone or receiving special stain or recut requests, or both, will have lower productivity. The most useful standard for employees in a given laboratory is set by the supervisor or senior technologists, or both, based on past productivity levels. To address productivity in a more global sense, it is necessary to assign work units to each of the varied tasks in histology, including loading and maintaining processors, embedding, cutting, routine staining, special stains, and immunohistochemistry. It is then possible to benchmark units worked per hour. As with routine cutting, however, the assignment of work unit values to a given task can only be done realistically by the histology supervisor and pathologist at a given site, taking into account economies of scale and levels of automation. Richard W. Brown, MD Medical Director, Core Histology Laboratory Memorial Hermann Healthcare System Houston Member, CAP Surgical Pathology Committee With members of the CAP/National Society for Histology Committee: Freida L. Carson, PhD, HT(ASCP) Lena T. Spencer, MA, HT(ASCP)HTL, QIHC Vincent Della Speranza, MMS, HT(ASCP)HTL Sue E. Lewis, HTL(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From relia1 <@t> earthlink.net Thu Aug 11 10:34:11 2005 From: relia1 <@t> earthlink.net (Pam Barker) Date: Fri Sep 16 15:25:26 2005 Subject: [Histonet] Job Alert from Pam Barker at RELIA Message-ID: Hi Histonetters, This is a job alert from Pam Barker at Relia. I have an immediate opening for a histo tech with one of my best clients. They are located in Northern Indiana, conveniently located close to Chicago and the suburbs. This is a private lab that supports several hospitals in Indiana and Illinois. They are growing and offer excellent compensation, benefits, relocation and opportunity for advancement. They also offer a great staff to work with. ASCP HT, HTL or registry eligible. This is a great opportunity for an experienced histo tech or a junior histo tech. If you are interested in hearing more about this position please contact me at 866-607-3542 or relia1@earthlink.net If you know of anyone else who might be interested please feel free to pass my information along to them as well. Thanks-Pam Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net From funderwood <@t> mcohio.org Thu Aug 11 11:09:50 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] VIP cleaning cycle Message-ID: Hi in Hisotland, This question is for those with a VIP 5 processor. Has anyone tried to get more than the recommended 5 runs from the cleaning xylene and alcohol? Thanks in adavance, Fred Underwood Mont. Co. Coroner Dayton, OH From jwatson <@t> gnf.org Thu Aug 11 11:27:56 2005 From: jwatson <@t> gnf.org (James Watson) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] FW: ASCP ACTION ALERT: Advocacy Needed for AFIP!! Message-ID: -----Original Message----- From: American Society for Clinical Pathology [mailto:Broadcast@ascp.org] Sent: Wednesday, August 10, 2005 10:22 AM To: James Watson Subject: ASCP ACTION ALERT: Advocacy Needed for AFIP!! ASCP Action Alert: Advocacy Needed for AFIP!! Marion James Watson; [Your ASCP ID is: 00973405]: The Armed Forces Institute of Pathology (AFIP) has been placed on the Department of Defense's most recent base closure list. ASCP is concerned about losing the future of the tissue repository and the valuable training and education services that the AFIP provides. Over 7,400 medical and other health professionals were educated and trained in 2004. Additionally, Each year the AFIP issues hundreds of thousands of hours in continuing medical education to a diverse group of medical professionals that specialize in pathology, forensics, radiology, emerging infectious disease, renal disease, ophthalmology, and urology. At a minimum, the AFIP tissue repository must remain accessible to experts in laboratory medicine. It would take five of the nine commissioners to remove a base from the list. The nine-member BRAC must send its recommendations on closures and realignments to the president by September 8, 2005. We need ASCP members to join us in the fight to save this valuable resource. Contact the BRAC and advoc ate for the removal of AFIP from the list of base closures. Follow this link to provide commentary to the BRAC commissioners. ASCP has included a sample letter with this message. You may use this letter by cutting and pasting the entire letter, or by cutting portions of the letter to supplement your personal information. Any personal insight that you provide will only aid in our effort. Please write today! http://www.brac.gov/feedback.aspx. Sample letter: The Honorable Anthony J. Principi Chairman Base Realignment and Closure Commission (BRAC) 2521 South Clark Street, Suite 600 Arlington, VA 22202 Dear Chairman Principi, I am writing to urge the BRAC to remove the Armed Forces Institute of Pathology (AFIP) from the list of military facilities slated for closure. The AFIP is an irreplaceable resource for disease research and patient care. This exceptional institution not only provides outstanding service to our nation's military community, but also contributes significant knowledge and education to the broader medical field's understanding of human disease. The AFIP tissue repository is a valuable national treasure that must remain accessible to researchers and experts in laboratory medicine. The AFIP's collection of specimens has helped medical professionals understand, develop vaccines and treatments for, and sometimes cure, often-fatal diseases. Access to both archived material as well as newly acquired specimens must be preserved. Each year the AFIP issues hundreds of thousands of hours in continuing medical education to a diverse group of medical professionals that specialize in pathology, forensics, radiology, emerging infectious disease, renal disease, ophthalmology, and urology. In fact, over 7,400 medical and other health professionals alone were educated and trained in 2004 through a variety of AFIP-sponsored venues including live courses, seminars and Internet courses. The AFIP's leadership in the area of education and training advances the knowledge base and ensures the competence of thousands of medical professionals. It would be a major loss to both military and civilian medicine, especially at this time, if AFIP's consultative services were discontinued. Should the AFIP be forced to close, our nation would lose a valuable resource that plays a key role in the advancement of medicine and contributes knowledge to improve the health of our nation's citizens. First, I respectfully request, that the BRAC eliminate its recommendation to dismantle the AFIP. Second, I urge the BRAC to ensure that the integrity of the tissue repository is preserved. Thank you for your kind consideration. Sincerely, [Your name here] Tell a friend: Not everyone receives ASCP's Action Alerts, so please forward this message to your peers and co-workers! ASCP America's Health Depends on Its Laboratories (c) 2003 American Society for Clinical Pathology 2100 West Harrison Street, Chicago, IL 60612-3798 312.738.1336 ~ info@ascp.org ABOUT THIS MESSAGE You are receiving this email because you are a member of ASCP, have attended ASCP programs/meetings or made a purchase from ASCP. If you no longer wish to receive emails of this kind from ASCP, please login to change your email preferences at ascp.org. This email message complies with all CAN-SPAM 2004 regulations. It was sent to you by the AMERICAN SOCIETY FOR CLINICAL PATHOLOGY, 2100 W. Harrison Street, Chicago, IL 60612 From ploykasek <@t> phenopath.com Thu Aug 11 11:37:48 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] H. Pylori IHC In-Reply-To: <9f.64cff233.302c1650@aol.com> Message-ID: Steve, What pretreatment are you using? Is your detection system avidin-biotin based or a polymer? What is the concentration of primary antibody? Does your negative exhibit any of this background staining? Sorry for all the questions, but I can help more once I know the answers. This is not a problem we've seen with the antibody we use. Patti Loykasek, BS,HTL, QIHC PhenoPath Laboratories Seattle, WA I have been getting a lot of background in the mucosa of the Gastric > biopsies, especially in cases that show increased inflammation and many > eosinophils. I > have increased the incubation time in peroxide blocker and protein blocker > and decreased the time in the primary and secondary Ab, only to find it > getting > worse instead of better. I am using Biocare's antibody and detection system on > their instrument. Any suggestions to eliminate this nonspecific staining? > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From katherine-walters <@t> uiowa.edu Thu Aug 11 12:27:43 2005 From: katherine-walters <@t> uiowa.edu (Walters, Katherine S) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] processing with glycol ether Message-ID: Hi All, A few years back I attended a workshop at NSH by Richard Dapson and Ada Feldman of Anatech. They used a glycol ether in place of ETOH for dehydrating tissue, as it is not supposed to harden tissue. I wonder if anyone out there in histo-land uses this routinely? Is it really more costly in the long run? I would also like to hear from Richard or Ada on this subject, as I attended this workshop in 1998 in Salt Lake City. They may have changed their opinion as well. Thanks for any input, Kathy Walters From PMonfils <@t> Lifespan.org Thu Aug 11 12:39:27 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] VIP cleaning cycle Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175A7@lsexch.lsmaster.lifespan.org> I just change the cleaning solvents when I change the processing solvents. I discard the cleaning solvents, and also the first absolute alcohol and the first xylene of the processing series. I use the second xylene and absolute alcohol of the processing series to replace the cleaning solvents. Then I move the third xylene and absolute up to the first position, and replace the later solvents with fresh reagent. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Fred > Underwood > Sent: Thursday, August 11, 2005 9:09 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] VIP cleaning cycle > > Hi in Hisotland, > > This question is for those with a VIP 5 processor. Has anyone tried to > get more than the recommended 5 runs from the cleaning xylene and > alcohol? > > Thanks in adavance, > > Fred Underwood > Mont. Co. Coroner > Dayton, OH > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From kgrobert <@t> rci.rutgers.edu Thu Aug 11 12:52:42 2005 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] VIP cleaning cycle In-Reply-To: References: Message-ID: <42FB906A.2020609@rci.rutgers.edu> No, I have not tried to stretch it, so I too would be interested if anyone did this as well. Kathleen Roberts Fred Underwood wrote: >Hi in Hisotland, > >This question is for those with a VIP 5 processor. Has anyone tried to >get more than the recommended 5 runs from the cleaning xylene and >alcohol? > >Thanks in adavance, > >Fred Underwood >Mont. Co. Coroner >Dayton, OH > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Janet.Bonner <@t> FLHOSP.ORG Thu Aug 11 13:18:49 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Cryostat sectioning of mammary gland Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB436E@fh2k093.fhmis.net> The Leica cryostats we use(CM1900) have a "snowflake" button to bring the temperature of the chuck holder to -50 degrees real quick. It is very useful when we get specimens like these! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Tessa Murray; histonet@lists.utsouthwestern.edu Sent: 8/11/2005 10:21 AM Subject: RE: [Histonet] Cryostat sectioning of mammary gland Dear Tess, Are you sure the cryochamber is below -30?C or just the specimen is at that temperature?? As your problem indicates that the knife & anti-roll plate are too warm, if the specimen is at -30?C. With the correct temperatures, mammary gland should be easy to section. 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: Tessa Murray [mailto:tessajmurray@hotmail.com] Sent: 10 August 2005 21:50 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cryostat sectioning of mammary gland I'm looking for some advice on cryosections. We are trying to collect frozen sections of mammary gland tissue and are experiencing problems - we do a lot of paraffin work here so the cryostat technique is quite new to us. We find that the knife cuts the OCT fine, but when it hits tissue it does not cut so we get sections with tissue shaped holes in them. We've tried changing the block and knife temp (we have tried altering them between -30 to -35C) but nothing seems to help. Any advice appreciated... tess _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Thu Aug 11 13:41:19 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] good altas for skin anatomy, etc. Message-ID: <000501c59ea4$43756780$a7d48a80@AMY> Does anyone know of a good atlas for skin anatomy. Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 liz <@t> premierlab.com Thu Aug 11 13:50:41 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] skin atlas Message-ID: <000f01c59ea5$9264d000$a7d48a80@AMY> Gina Human skin. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 info <@t> instrumedics.com Thu Aug 11 13:54:52 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] cutting mammary gland Message-ID: <004b01c59ea6$483c2c70$6401a8c0@INSTRUMEDICS22> Tessa, It is very important to snap freeze the tissue to minimize the ice crystal growth which can damage the cells. We recommend the Gentle Jane method where you get good thermal exchange with the liquid nitrogen chilled heat extractor. Both low temperature and thermal exchange improve freezing. The CryoJane Tape-Transfer system makes it easy to cut a frozen section whether it is fatty tissue or hard tissue. It can be cut as thin as 2-4 microns. The section is captured on a cold tape, flat and uncompressed, and then transferred to a cold slide. The section remains frozen until it immersed in a fixative or freeze-substituted. The morphology is generally remarkable. Melting the frozen section when mounting on a room temperature slide degrades the quality. Please go to www.instrumedics.com for more details. Click on the "gallery" to see photomicrographs of CryoJane prepared frozen sections. We welcome your questions. Bernice From MadaryJ <@t> MedImmune.com Thu Aug 11 15:34:15 2005 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] You crazy histotechs Message-ID: <746FDB897740814EA52BDDCB5ED1DDBCBB1829@medimmune4.medimmune.com> Pam and Tim among others are officially my newest heroes. I guess I can agree with most, but will say that I am not the histotech today that I was over a quarter of a century ago which is why I have to overlook some of the stuff I see going away that should stay regarding training. I also can say that the alphabet soup after our names(at least for me) was only to get or maintain a job, I never felt like I had the time to do them, and it has always been this ego thing attached to it. I certainly wasn;t any better after I did it since it was stuff I did already. I mean we all know of techs out there that are not certified that rock, and vice versa. I just feel bad that there is such a disparity in pay all of the country, I have been under and overpaid over the years quite honestly. Approaching Retirement Again in Maryland, Joseph "Nick" Madary HT, HTL(ASCP)QIHC Lab Mgr, Medimmune Inc Gaithersburg, MD 20878 301.398.4745 301.397.9745(fax) From JMahoney <@t> alegent.org Thu Aug 11 15:48:59 2005 From: JMahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] HT Workforce Message-ID: Hello, Does anyone know what the average age of registered Histo techs is? How many retire each year and how many new ones come into the field? Thanks for the help. Jan Omaha Janice Mahoney Histology/Cytology Coordinator Alegent Health Laboratory Phone (402)717-2889 Fax (402)717-5231 "We pledge to be creative, visionary leaders committed to holistic healthcare in the region" From gliuygao <@t> hotmail.com Thu Aug 11 16:04:01 2005 From: gliuygao <@t> hotmail.com (yan gao) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] (no subject) Message-ID: Dear histonet, I have a question for you. I am staining MAP kinase in tumor tissue. It is a Cytoplasma stain. The stain is not very stable. Sometime strong and sometime weak. How can I let protein stay in cytoplasma? Not run away? Any idea? Best Regards, Yan From gliuygao <@t> hotmail.com Thu Aug 11 16:04:01 2005 From: gliuygao <@t> hotmail.com (yan gao) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] (no subject) Message-ID: Dear histonet, I have a question for you. I am staining MAP kinase in tumor tissue. It is a Cytoplasma stain. The stain is not very stable. Sometime strong and sometime weak. How can I let protein stay in cytoplasma? Not run away? Any idea? Best Regards, Yan From PMonfils <@t> Lifespan.org Thu Aug 11 16:07:18 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] You crazy histotechs Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175A8@lsexch.lsmaster.lifespan.org> Then again, there are certfied pathologists and surgeons who are outstanding, and there are certified pathologists and surgeons who are mediocre or worse. But if I was the patient, I would still feel pretty strongly about having certified doctors handling my case. And certified lab personnel. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Madary, Joseph > Sent: Thursday, August 11, 2005 1:34 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] You crazy histotechs > > Pam and Tim among others are officially my newest heroes. I guess I can > agree with most, but will say that I am not the histotech today that I was > over a quarter of a century ago which is why I have to overlook some of > the stuff I see going away that should stay regarding training. I also > can say that the alphabet soup after our names(at least for me) was only > to get or maintain a job, I never felt like I had the time to do them, and > it has always been this ego thing attached to it. I certainly wasn;t any > better after I did it since it was stuff I did already. I mean we all know > of techs out there that are not certified that rock, and vice versa. I > just feel bad that there is such a disparity in pay all of the country, I > have been under and overpaid over the years quite honestly. > > Approaching Retirement Again in Maryland, > > Joseph "Nick" Madary HT, HTL(ASCP)QIHC > Lab Mgr, Medimmune Inc > Gaithersburg, MD 20878 > 301.398.4745 > 301.397.9745(fax) > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Eric <@t> ategra.com Thu Aug 11 17:33:33 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Histo Tech Mananger Supervisor, Bench Tech's and Temps needed for immediate openings (this is not a duplicate) Message-ID: Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisor, Histo Managers and Histo Bench Techs. These are fulltime permanent positions. Right now my Hottest jobs are in Colorado and Oregon who are seeking Lab Managers and Lab Supervisor. I have positions in California and Michigan who are seeking Histotech Supervisors. I also have Histo bench positions in Colorado,California,Pennsylvania,Illinois,New Hampshire,Michigan, Georgia,Florida,South Carolina, and Massachusetts. I also have a traveler positions available in South Carolina at a Dermapathology lab. The assignment would be for 1 to 2 months and you must have dermpath experience. I have Temp assignment openings in South Florida, and California. There are also Research openings in Massachusetts and Ohio as well The clients are currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recriuter 1-800-466-9919 ext 223 From Jacqueline.Malam <@t> rli.mbht.nhs.uk Fri Aug 12 06:43:22 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Fridge digital thermometers Message-ID: Does anyone know where I can find a supplier in Britain for a digital thermometer suitable for a fridge or freezer, which will also have an audible alarm when the temperature is outside set limits, preferably something situated outside the fridge and not too expensive. Jacqui Malam Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From sjchtascp <@t> yahoo.com Fri Aug 12 09:16:31 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] skin control block Message-ID: <20050812141631.62344.qmail@web90203.mail.scd.yahoo.com> I work in animal research and am in need of a good human skin paraffin control block as a testing control of the Pinkus elastic stain. Thanks, Steve --------------------------------- Start your day with Yahoo! - make it your home page From donald <@t> labcareer.com Fri Aug 12 09:26:10 2005 From: donald <@t> labcareer.com (Donald Knowles) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Histo positions on West Coast of Florida Message-ID: <26A7728EF768DA478F45954252293C202E54B9@matrix.HCCI.local> Hi , I currently have a Hospital on the West Coast of Florida that is desperate for 2 Histotechs. Please if anyone wants to escape the pending frost of winter and is willing to get a Florida license please see www.doh.state.fl.us/mqu and lets talk ! I also pay referral bonuses if the person you recommend is placed through my company. Thanks for your time. Don Knowles Healthcare Connections Inc. 866-346-8522 ext 311 From Heather.A.Harper <@t> pcola.med.navy.mil Fri Aug 12 10:56:16 2005 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] CAP..Need info Message-ID: <807FE48C5A7CC940B973B58D32E701431A850BE0@nhpens-exch1.pcola.med.navy.mil> Our hospital did the histology CAP, where you send in the requested slides, and you can earn CE points by doing the final critique. I want and need to know, when they send you back your results, is there any forms you have to fill out like what corrective action was taken etc.. Since this is something new that CAP has started, we are not sure what our next move is and I have already received the next CAP packet and must have slides submitted no later than Sept. 2 Also to get credit for the Critique, do I need to create a password and log onto the CAP website? Any info is very much appreciated. Heather A. Harper Histology Supervisor Naval Hospital of Pensacola, FL From SB_Cohen <@t> fccc.edu Fri Aug 12 11:44:33 2005 From: SB_Cohen <@t> fccc.edu (Cohen, Sherene B.) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Extended Slide and Block Storage Message-ID: Hello all, Our lab is unique that all slides and blocks are retained indefinitely. While we do use offsite storage facilities, this is becoming expensive and unreliable. A compressed storage system was proposed. I was wondering if anyone else has similar problems or has employed alternative storage solutions. From Laurie.Pereira <@t> sdcounty.ca.gov Fri Aug 12 12:01:16 2005 From: Laurie.Pereira <@t> sdcounty.ca.gov (Pereira, Laurie ) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] rat heart cryos' for immunos Message-ID: Does anyone know why heart cryos tend to separate? We've tried fresh frozen, NBF perfused, sucrose perfused and FBS perfused and the heart tissue has 'holes' in it. Laurie L. Pereira, RVT SDCADDL (County Veterinarian) 5555 Overland Avenue Suite 4103 San Diego, CA. 92123 Phone: 858-694-3384 Fax: 858-571-4268 From doninn <@t> mail.nih.gov Fri Aug 12 12:17:25 2005 From: doninn <@t> mail.nih.gov (Donin, Nick (NIH/NCI)) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Human Specific Ab in FFPE mouse tissue Message-ID: <16A0583FB1644E4DB8C0A0265028B6FD02B4E7CD@nihexchange13.nih.gov> Hello, I'm attempting to stain xenografted human cells in mouse brain and am not having much success. The cells are glioma U251 cells expressing GFP, and I'd like to stain for both GFP and for all human cells. We tried once already with two antibodies and both sets came out completely clean, no staining. We know, at least, that our secondary, anti-mouse antibody does not bind non-specifically to our mouse tissue, which is good. Can someone suggest a human specific antibody or antigen, and also an anti-GFP antibody that works reliably in FFPE tissue. My protocol is below. Please email me at doninn@mail.nih.gov with any suggestions. Thanks so much everyone, it is much appreciated. Nick. PARAFFIN SECTIONS (fixed using PFA) 1. 2 X Xylene, 5 min 2. 2 X 100% ETOH, 5 min 3. 2 X 95% ETOH, 5 min 4. 1 X 70% ETOH, 5 min 5. Proceed to step 1 for Frozen Sections OR go to Antigen Retrieval ANTIGEN RETRIEVAL 1. Heat Vector Antigen retrieval solution (working dilution) in pressure cooker 2. Immerse slides, close top, and heat until cap begins rocking 3. Allow to rock for 1 minute 4. Take cooker off heat source, allow to cool, remove slides (Go to step two of Frozen Sections) FROZEN SECTIONS 1. Fix 12 min in 85% ETOH 2. Wash PBS 3 x 5 min (or 3x10 dips) 3. Quench 20 min. in 3% hydrogen peroxide 4. Wash PBS 3 x 5 min 5. Block 1 hr (or 20 min) in PBS + 10% Horse serum (10% frozen serum in PBS) 6. Add 1? antibody diluted in PBS + 2% NHS, 4? overnight, in a humidified chamber (start with double concentration recommended for Western, 200 ul per slide. NEXT DAY 1. Washin PBS 3 x 5 min 2. Add 2? Antibody: Vector kit: 2 drops (100 ul) NHS, 2 drops (100 ul) antibody in 5 ml PBS, 1 hr (or 30 min) at RT. (Make up ABC during this incubation - Vector Kit: 2 drops A + 2 drops B in 5 ml PBS... must sit at least 30 min at RT) 3. Wash PBS 3 x 5 min 4. Add ABC, 1 hr (or 30 min) at RT 5. Wash PBS 3 x 5 min 6. Add DAB: Vector Kit: 2 drops buffer, 4 drops DAB, 2 drops H202 in 5 ml DDH20). Leave 30 sec. to 5 min depending on color development (monitor visually for color change). 7. Stop in tap water 8. Counterstain in Hemotoxylin (Mayers - 10 min) 9. Wash with running tap water 2-5 min (longer washing makes stain more blue) 10. Dip in 95% ETOH 11. 2 x 100% ETOH 2 min 12. 2 x Xylene 2 min 13. Permount Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 From aep10 <@t> cornell.edu Fri Aug 12 12:19:49 2005 From: aep10 <@t> cornell.edu (Anna Elisse Beaudin) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] beta-galactosidase in mouse embryos Message-ID: <1817.128.253.96.73.1123867189.squirrel@128.253.96.73> Hi all, I am planning to do beta-galactosidase staining on dpc 9.5 mouse embryos, and I was wondering if anyone had any advice on the best fixative to use prior to staining, as well as the best clearing agent to use after. I've seen many protocols that call for one hour 4% paraformaldehyde fix, but when I have done Bgal stain on adult tissue and tissue sections, this fix seems to reduce staining. I was also planning to use 100% methyl salicylate as a clearing agent. I'd very much appreciate anyone's advice! thanks in advance, Anna Beaudin Division of Nutritional Sciences Cornell University From kbroomal <@t> NEMOURS.ORG Fri Aug 12 12:23:36 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] rat heart cryos' for immunos Message-ID: <6E41111281623B4B8A9AB8F9A7EA34378243D2@wlmmsx02.nemours.org> How are you freezing your tissue? It's probably freezing artifact. Does it look like big donut holes in the middle of the muscle fibers? Kristen Broomall -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Pereira, Laurie Sent: Friday, August 12, 2005 1:01 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] rat heart cryos' for immunos Does anyone know why heart cryos tend to separate? We've tried fresh frozen, NBF perfused, sucrose perfused and FBS perfused and the heart tissue has 'holes' in it. Laurie L. Pereira, RVT SDCADDL (County Veterinarian) 5555 Overland Avenue Suite 4103 San Diego, CA. 92123 Phone: 858-694-3384 Fax: 858-571-4268 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Laurie.Pereira <@t> sdcounty.ca.gov Fri Aug 12 12:49:29 2005 From: Laurie.Pereira <@t> sdcounty.ca.gov (Pereira, Laurie ) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] rat heart cryos' for immunos Message-ID: We are using isopentane and it takes about a minute to freeze the tissue. There appears to be cracks between the muscle fibers. -----Original Message----- From: Kristen Broomall [mailto:kbroomal@NEMOURS.ORG] Sent: Friday, August 12, 2005 10:24 AM To: Pereira, Laurie ; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] rat heart cryos' for immunos How are you freezing your tissue? It's probably freezing artifact. Does it look like big donut holes in the middle of the muscle fibers? Kristen Broomall -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Pereira, Laurie Sent: Friday, August 12, 2005 1:01 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] rat heart cryos' for immunos Does anyone know why heart cryos tend to separate? We've tried fresh frozen, NBF perfused, sucrose perfused and FBS perfused and the heart tissue has 'holes' in it. Laurie L. Pereira, RVT SDCADDL (County Veterinarian) 5555 Overland Avenue Suite 4103 San Diego, CA. 92123 Phone: 858-694-3384 Fax: 858-571-4268 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ynwang <@t> u.washington.edu Fri Aug 12 13:09:06 2005 From: ynwang <@t> u.washington.edu (Y. Wang) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] cryojane system inquiry Message-ID: Dear all, We want to purchase a cryojane system for our frozen tissue (we have tried one and it works really well). However, in order to satisfy the requirements of our purchasing department I need to show them that I have made a 'thorough inquiry' as to if there is another system out there that can do this job and why the cryojane is better for our application. From my searches on the web and from the histonet e-mails I have not seen any other system that can do what the cryojane can do for us (make thin sections from frozen tissue with implants in them without the implant falling out while sectioning). However, it is possible I may be missing something, if so, please let me know. However, if I am correct in my thinking also please let me know so I can add 'query to a professional list server' as one of my search areas (as I was given the impression web searches aren't enough!). Thank you for your help Yak-Nam Senior Fellow Department of Bioengineering University of Washington Box 357962 Seattle, WA 98195 Tel.: (206)-221-5873 Fax.: (206)-221-5874 From TJJ <@t> Stowers-Institute.org Fri Aug 12 13:12:37 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Re: HT Workforce Message-ID: Good luck getting that information from a group of (mostly) women. :-) Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org You wrote: Hello, Does anyone know what the average age of registered Histo techs is? How many retire each year and how many new ones come into the field? Thanks for the help. Jan Omaha Janice Mahoney Histology/Cytology Coordinator Alegent Health Laboratory Phone (402)717-2889 Fax (402)717-5231 "We pledge to be creative, visionary leaders committed to holistic healthcare in the region" From Thomas.Crowell <@t> biogenidec.com Fri Aug 12 13:21:19 2005 From: Thomas.Crowell <@t> biogenidec.com (Thomas Crowell) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Myelin Basic Protein In-Reply-To: Message-ID: Is anyone using an antibody to MBP on FFPE mouse tissues? Thanks Tom Crowell BiogenIdec Cambridge, MA From dmarsha3 <@t> utmem.edu Fri Aug 12 13:48:27 2005 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] cryojane system inquiry References: Message-ID: <000301c59f6e$6d885f80$f623c084@DanaM> Hi, I went through the exact same thing myself, (federal lab). There is nothing that i could find anywhere. We have a cryojane and are happy with it. Dana Marshall ----- Original Message ----- From: "Y. Wang" To: Sent: Friday, August 12, 2005 1:09 PM Subject: [Histonet] cryojane system inquiry > Dear all, > > We want to purchase a cryojane system for our frozen tissue (we have tried > one and it works really well). However, in order to satisfy the > requirements of our purchasing department I need to show them that I have > made a 'thorough inquiry' as to if there is another system out there that > can do this job and why the cryojane is better for our application. From > my searches on the web and from the histonet e-mails I have not seen any > other system that can do what the cryojane can do for us (make thin > sections from frozen tissue with implants in them without the implant > falling out while sectioning). However, it is possible I may be missing > something, if so, please let me know. However, if I am correct in my > thinking also please let me know so I can add 'query to a professional > list server' as one of my search areas (as I was given the impression web > searches aren't enough!). > > Thank you for your help > Yak-Nam > > > 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 jennifer.l.hofecker <@t> Vanderbilt.Edu Fri Aug 12 14:04:02 2005 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] problems with Gomori's Trichrome on Frozen Muscles Message-ID: <898D946569A27444B65667A49C07405207545C@mailbe06.mc.vanderbilt.edu> Happy Friday everyone, We are experiencing a strange phenomenon with our Gomori's. All of a sudden the staining is predominately red (instead of green). Normal muscle control is still green but the patient tissues are staining red. We have tried several different changes: new trichrome solution, recutting frozens, etc. The only thing that is reproducible is that sections which have been previously cut and stored at -70 seem to stain fine. If we cut a frozen then begin staining it without time in the freezer, voila, the same muscle sections are now red instead of green. What could be causing this? I even pulled out the same piece of control tissue and cut fresh slides on it. They were red also. There was one fresh slide that stained green in the whole "experiment" otherwise, anything that doesn't spend time in -70 is turning red. We are using the same procedure that we have been using for quite a while, and the precut slides do stain, so I don't think it's a procedural error. The only change is that we have been trying to stain the trichromes when the specimens are cut instead of storing in the freezer to batch staining later. Has anybody experienced this? Any ideas would be appreciated. Thanks in advance, Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax From vazquezr <@t> ohsu.edu Fri Aug 12 09:11:42 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Fridge digital thermometers Message-ID: Hello, Fisher Scientific should have them. Robyn OHSU >>> "Malam Jacqueline" 8/12/2005 4:43 AM >>> Does anyone know where I can find a supplier in Britain for a digital thermometer suitable for a fridge or freezer, which will also have an audible alarm when the temperature is outside set limits, preferably something situated outside the fridge and not too expensive. Jacqui Malam Lancaster England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Fri Aug 12 14:35:39 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Fast Myosin Message-ID: Thanks to all of you who responded to my question about IHC staining for "fast myosin". The antibody from Sigma is fabulous! We get incredible reactivity using a 1:10,000 dilution for 30 minutes with "no" pretreatment. Thanks again! 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 ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From BDUE <@t> PARTNERS.ORG Fri Aug 12 14:45:44 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] problems with Gomori's Trichrome on Frozen Muscles Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB501027110@PHSXMB7.partners.org> Hello Jennifer, I have noticed two things which cause red staining. a) incomplete rinse after hematoxylin. If the section is wrinkly or puckered and traps the hematoxylin soln, those areas tend to stain red with the gomori. Longer rinse after hematoxylin reduces this. This is probably not your trouble, but you could try extending your rinse before the trichrome soln. b) "freeze-dried-looking" regions of frozen sections. Sometimes two consecutive sections will look different after they've dried on the slides. Thid can be immediately obvious in the cryostat, but often shows up only after storage at -80. The "good" section will look translucent, while the "bad" one will have areas that look opaque, white and frosty -- as if it had freeze dried, or as if it hadn't really stuck to the slide fully, or something. Those white areas tend to show funky staining -- perhaps for the same reason as above: they may trap solns more than usual. Are you picking up sections exactly the same way or handling them differently? I know you said you are staining same day. I have cut and stained 30min later without problems. c) another remote possibility is that the sections are getting exposed to some solvent or formalin vapors at some point. Fixation and/or lipid extraction can ruin gomori staining, although I have usually seen an absence of red rather than excess. Curious to know what the problem is, -brice Neuropathology LAb Brigham & Women's Hospital Boston "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Hofecker, Jennifer L Sent: Friday, August 12, 2005 3:04 PM To: histonet Subject: [Histonet] problems with Gomori's Trichrome on Frozen Muscles Happy Friday everyone, We are experiencing a strange phenomenon with our Gomori's. All of a sudden the staining is predominately red (instead of green). Normal muscle control is still green but the patient tissues are staining red. We have tried several different changes: new trichrome solution, recutting frozens, etc. The only thing that is reproducible is that sections which have been previously cut and stored at -70 seem to stain fine. If we cut a frozen then begin staining it without time in the freezer, voila, the same muscle sections are now red instead of green. What could be causing this? I even pulled out the same piece of control tissue and cut fresh slides on it. They were red also. There was one fresh slide that stained green in the whole "experiment" otherwise, anything that doesn't spend time in -70 is turning red. We are using the same procedure that we have been using for quite a while, and the precut slides do stain, so I don't think it's a procedural error. The only change is that we have been trying to stain the trichromes when the specimens are cut instead of storing in the freezer to batch staining later. Has anybody experienced this? Any ideas would be appreciated. Thanks in advance, Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From contact <@t> excaliburpathology.com Fri Aug 12 14:53:21 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Problems with Gomori's Trichrome on Frozen Muscle Message-ID: <20050812195321.41660.qmail@web50302.mail.yahoo.com> Results for Gomori's Trichrome Stain: Cell cytoplasm, MUSCLE - Red Collagen - Green Nuclei - Blue to Black Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From kbroomal <@t> NEMOURS.ORG Fri Aug 12 15:09:33 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Problems with Gomori's Trichrome on Frozen Muscle Message-ID: <6E41111281623B4B8A9AB8F9A7EA34378243D4@wlmmsx02.nemours.org> Paula, It is supposed to be greenish. Modified Gomori's Trichrome method for muscle biopsies: Nuclei - red/purple Normal Muscle myofibrils - green with distinct A & I bands Intermyofibrillar muscle - red Interstitial collagen - green Jennifer, We just got a weird red artifact (big splotch of red in the middle of the section) the other week on a biopsy, but it had been cut on another cryostat other than ours. We thought it may have been due to a dull blade or something like that. We recut it on our cryo & it stained fine. I hope you get it worked out & I'm interested to know what's going on! Good luck. Kristen Broomall, HT (ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Paula Pierce Sent: Friday, August 12, 2005 3:53 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Problems with Gomori's Trichrome on Frozen Muscle Results for Gomori's Trichrome Stain: Cell cytoplasm, MUSCLE - Red Collagen - Green Nuclei - Blue to Black Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Fri Aug 12 15:11:26 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] problems with Gomori's Trichrome on Frozen Muscles References: <898D946569A27444B65667A49C07405207545C@mailbe06.mc.vanderbilt.edu> Message-ID: <42FD026D.C53C036B@uwo.ca> If the method is Gomori's one-step trichrome, with chromotrope 2R and light green (or fast green FCF), and PTA or PMA, then muscle should stain red, and collagen green. Instructions for 2 variants of the method are given in Presnell & Schreibman (1997) Humason's Animal Tissue Techniques, 5th edn. Baltimore: Johns Hopkins Univ. Press, pp.129-131. I think it's also in earlier editions of the book (with G. Humason as the author). And in other books too, of course. Trichrome methods are usually done on paraffin rather than frozen sections. If the fixative is formaldehyde, staining is improved by a pre-treatment of the hydrated sections with picric acid - Bouin's solution is often used. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Hofecker, Jennifer L" wrote: > > Happy Friday everyone, > We are experiencing a strange phenomenon with our Gomori's. All of a sudden the staining is predominately red (instead of green). Normal muscle control is still green but the patient tissues are staining red. We have tried several different changes: new trichrome solution, recutting frozens, etc. The only thing that is reproducible is that sections which have been previously cut and stored at -70 seem to stain fine. If we cut a frozen then begin staining it without time in the freezer, voila, the same muscle sections are now red instead of green. What could be causing this? I even pulled out the same piece of control tissue and cut fresh slides on it. They were red also. There was one fresh slide that stained green in the whole "experiment" otherwise, anything that doesn't spend time in -70 is turning red. We are using the same procedure that we have been using for quite a while, and the precut slides do stain, so I don't think it's a procedural error. The only change > is that we have been trying to stain the trichromes when the specimens are cut instead of storing in the freezer to batch staining later. Has anybody experienced this? Any ideas would be appreciated. > > Thanks in advance, > Jennifer > > > > > Jennifer Hofecker, HT (ASCP) > Vanderbilt University Medical Center > Division of Neuropathology > (615) 343-0083 > (615) 343-7089 fax > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ljb <@t> medicine.wisc.edu Fri Aug 12 15:17:08 2005 From: ljb <@t> medicine.wisc.edu (LaCinda Burchell) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Problems with Gomori's Trichrome on Frozen Muscle Message-ID: Easy there, come on , it was a well stated and honest question. Play nice. >>> Paula Pierce 08/12/05 2:53 PM >>> Results for Gomori's Trichrome Stain: Cell cytoplasm, MUSCLE - Red Collagen - Green Nuclei - Blue to Black Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Aug 12 15:18:00 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Flourescent labels Message-ID: <200508122017.j7CKHvfw004382@chip.viawest.net> Folks, I was under the general impression that most flourescent labels were done on frozen tissue because of problems with auto flourescence associated with aldehyde fixation and paraffin processing. Has the technology developed so that these new flourescent labels such as the Alexa dyes can now be used on ffpe tissues? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From tpmorken <@t> labvision.com Fri Aug 12 15:31:52 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Flourescent labels Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D002@usca0082k08.labvision.apogent.com> Patsy, The average routine fluorescent system uses "longpass" emission filters that let almost all the light from blue to red through. Therefore you will see many colors of autofluoresce. Since the autofluorescence is usually a different color than your target fluorchrome, the way you can avoid autofluoresence is to find filters that block the autofluorescent wavelength. It takes more effort to coordinate the different wavelengths associated with the fluorochrome excitation and emission vs the autofluorescent excitation and emission, but by using narrow-bandwidth filters (20-40 nm bandwidth) it is usually possible. Both Omega and Chroma have these filters. 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 Patsy Ruegg Sent: Friday, August 12, 2005 1:18 PM To: 'Histonet' Subject: [Histonet] Flourescent labels Folks, I was under the general impression that most flourescent labels were done on frozen tissue because of problems with auto flourescence associated with aldehyde fixation and paraffin processing. Has the technology developed so that these new flourescent labels such as the Alexa dyes can now be used on ffpe tissues? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NSTYLOPOULOS <@t> PARTNERS.ORG Fri Aug 12 16:04:02 2005 From: NSTYLOPOULOS <@t> PARTNERS.ORG (Stylopoulos, Nicholas) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Immunofluorescence problem-Non-specific binding appears in control slide Message-ID: <4063C6544431714FA0EDA3D9138CB7A4A81194@PHSXMB5.partners.org> Hello everybody and thank you for taking the time to read this message. I have been struggling with immunofluorescence and I have the following problem: My control slide (secondary antibody only) appears to have specific staining. I have repeated the assay many times (Also, I showed my control slide to many people and they asked me whether I am sure that I have not added my primary antibody as well!). My primary antibody is goat anti-rat (against chromogranin A). I have also tried rabbit anti-rat. My secondary is donkey anti-goat FITC conjugated (I have tried donkey anti-rabbit and goat anti-rabbit). Basically, I am trying to visualize the endocrine cells in the gastrointestinal tract. This is my protocol: Tissue: rat ileum-snap frozen in isopentane Sections: Frozen sections 3-5?m Fixation: 4% paraformaldehyde (I tried also acetone -20C from 30secs to 20min and it seems that acetone destroys the histology of my sections) Washx3 with filtered PBS 1X Triton x-100 0.05% for 5min Wash x1 with filtered PBS 1X Block with 10% donkey serum (I tried adding 10% rat serum or 3% BSA) for 2hrs (I tried also blocking overnight... or I added Background buster (from Inovvex) and Fc receptor blocker (from Innovex too) or sodium azide 0.1%) Primary Antibody in PBS for 1hr Washx3 with filtered PBS 1X Secondary antibody in PBS for 1 hr Washx6 with filtered PBS 1X Mount with DAPI Any suggestions will be extremely appreciated....Thanks again! Nick From la.sebree <@t> hosp.wisc.edu Fri Aug 12 16:11:25 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] anti-mitochondria antibody Message-ID: Hello histonetters, Our neuropathologist is looking for a reference lab, preferably one that will do "stain only", to get an anti-mitochondria stain done using clone M-II68. Thanks for your assistance, Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 From bwhitaker <@t> brownpathology.com Fri Aug 12 16:21:33 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] problems with Gomori's Trichrome on Frozen Muscles In-Reply-To: <42FD026D.C53C036B@uwo.ca> Message-ID: <000001c59f83$d0d02040$3601a8c0@brownpathology.net> Jennifer, If I understand correctly, you are probably doing frozen muscle biopsies for a neuropathologist as part of a whole muscle biopsy work-up. The Gomori's modification used by neuropathologists does indeed produce green muscle, if the muscle is normal. Only "ragged red fibers" should be staining red. I really don't know the specifics of this, but you don't do the bouins post-fixation. As I recall a friend and former co-worker had pH problems that caused some variances in her muscles w/ Gomori's. I have some muscle experience, but am not currently doing that, and don't have access to the references I used. Maybe someone with expertise in neuropath will address this issue with more clarity than I can. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Friday, August 12, 2005 3:11 PM To: Hofecker, Jennifer L Cc: histonet Subject: Re: [Histonet] problems with Gomori's Trichrome on Frozen Muscles If the method is Gomori's one-step trichrome, with chromotrope 2R and light green (or fast green FCF), and PTA or PMA, then muscle should stain red, and collagen green. Instructions for 2 variants of the method are given in Presnell & Schreibman (1997) Humason's Animal Tissue Techniques, 5th edn. Baltimore: Johns Hopkins Univ. Press, pp.129-131. I think it's also in earlier editions of the book (with G. Humason as the author). And in other books too, of course. Trichrome methods are usually done on paraffin rather than frozen sections. If the fixative is formaldehyde, staining is improved by a pre-treatment of the hydrated sections with picric acid - Bouin's solution is often used. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Hofecker, Jennifer L" wrote: > > Happy Friday everyone, > We are experiencing a strange phenomenon with our Gomori's. All of a sudden the staining is predominately red (instead of green). Normal muscle control is still green but the patient tissues are staining red. We have tried several different changes: new trichrome solution, recutting frozens, etc. The only thing that is reproducible is that sections which have been previously cut and stored at -70 seem to stain fine. If we cut a frozen then begin staining it without time in the freezer, voila, the same muscle sections are now red instead of green. What could be causing this? I even pulled out the same piece of control tissue and cut fresh slides on it. They were red also. There was one fresh slide that stained green in the whole "experiment" otherwise, anything that doesn't spend time in -70 is turning red. We are using the same procedure that we have been using for quite a while, and the precut slides do stain, so I don't think it's a procedural error. The only change > is that we have been trying to stain the trichromes when the specimens > are cut instead of storing in the freezer to batch staining later. > Has anybody experienced this? Any ideas would be appreciated. > > Thanks in advance, > Jennifer > > > > > Jennifer Hofecker, HT (ASCP) > Vanderbilt University Medical Center > Division of Neuropathology > (615) 343-0083 > (615) 343-7089 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 liz <@t> premierlab.com Fri Aug 12 16:52:23 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] eotaxin antibody Message-ID: <000501c59f88$1f1b2b10$a7d48a80@AMY> Hello everyone. Is there anyone out there that knows of an antibody to eotaxin that will work in ffpe tissues that is commercially available? I see lots of references for it on paraffin sections, but they have made up their own antibody. I have also seen that there are quite a few vendors that cell antibodies to extaxin but they don't say it works for IHC, but rather westerns, etc. Any help would be appreciated. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 histosci <@t> shentel.net Fri Aug 12 17:00:36 2005 From: histosci <@t> shentel.net (HSRL) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Leica TP-4040 Linear Stainer Message-ID: <000201c59f89$4b4cafa0$0200a8c0@HSRLMAIN> Hey Netters, Anyone have an opinion on the Leica TP-4040 Linear Stainer? We are in need of something faster than 120 H&E slides/hour and feel the Leica would be a good fit. Any input is greatly appreciated. -Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 5930 Main Street Mount Jackson, Virginia 22842 540.477.4440 540.477.4448 tomgalati@hsrl.org www.hsrl.org From BDUE <@t> PARTNERS.ORG Fri Aug 12 17:03:44 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] problems with Gomori's Trichrome on Frozen Muscles Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB501027111@PHSXMB7.partners.org> The Gomori used in muscle panels is the ENGEL-CUNNINGHAM MODIFICATION which adjusts the pH of the standard Gomori one-step. As far as I know it only works properly on fresh unfixed frozen sections. It is very sensitive to the condition of the tissue (fixation, lipid extraction, freeze quality, fresh bx drying, etc.). Online, see: http://www.neuro.wustl.edu/neuromuscular/pathol/histol/trichrom.htm -brice "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of John Kiernan Sent: Friday, August 12, 2005 4:11 PM To: Hofecker, Jennifer L Cc: histonet Subject: Re: [Histonet] problems with Gomori's Trichrome on Frozen Muscles If the method is Gomori's one-step trichrome, with chromotrope 2R and light green (or fast green FCF), and PTA or PMA, then muscle should stain red, and collagen green. Instructions for 2 variants of the method are given in Presnell & Schreibman (1997) Humason's Animal Tissue Techniques, 5th edn. Baltimore: Johns Hopkins Univ. Press, pp.129-131. I think it's also in earlier editions of the book (with G. Humason as the author). And in other books too, of course. Trichrome methods are usually done on paraffin rather than frozen sections. If the fixative is formaldehyde, staining is improved by a pre-treatment of the hydrated sections with picric acid - Bouin's solution is often used. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Hofecker, Jennifer L" wrote: > > Happy Friday everyone, > We are experiencing a strange phenomenon with our Gomori's. All of a sudden the staining is predominately red (instead of green). Normal muscle control is still green but the patient tissues are staining red. We have tried several different changes: new trichrome solution, recutting frozens, etc. The only thing that is reproducible is that sections which have been previously cut and stored at -70 seem to stain fine. If we cut a frozen then begin staining it without time in the freezer, voila, the same muscle sections are now red instead of green. What could be causing this? I even pulled out the same piece of control tissue and cut fresh slides on it. They were red also. There was one fresh slide that stained green in the whole "experiment" otherwise, anything that doesn't spend time in -70 is turning red. We are using the same procedure that we have been using for quite a while, and the precut slides do stain, so I don't think it's a procedural error. The only change > is that we have been trying to stain the trichromes when the specimens are cut instead of storing in the freezer to batch staining later. Has anybody experienced this? Any ideas would be appreciated. > > Thanks in advance, > Jennifer > > > > > Jennifer Hofecker, HT (ASCP) > Vanderbilt University Medical Center > Division of Neuropathology > (615) 343-0083 > (615) 343-7089 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 NBrandenburger <@t> Bloomhealth.org Fri Aug 12 17:40:02 2005 From: NBrandenburger <@t> Bloomhealth.org (Brandenburger, Nancy) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] COMPETENCY TESTING Message-ID: <2EB2FCDAF450DA4483CFFD56F30E0C17EA025C@EXCHVS1.neutzone.bloomhealth.org> When doing Competency Testing for our annual Performance Assessment, the job description is always incorporated. However, the system we use for testing of technical skills is somewhat cumbersome and time-consuming. I would appreciate any suggestions anyone has for completing this portion of the Performance Assessment. Thanks. Nancy Brandenburger HT (ASCP) LIS Specialist Histology Lead Tech phone: 812-353-5306 fax: 812-353-5584 CONFIDENTIALITY NOTICE: This e-mail message, including all attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. If you are not the intended recipient, you may NOT use, disclose, copy or disseminate this information. Please contact the sender by reply e-mail immediately and destroy all copies of the original message including all attachments. Your cooperation is greatly appreciated. Bloomington Hospital & Healthcare System P.O. Box 1149 Bloomington, IN 47402 From al.floyd <@t> juno.com Fri Aug 12 19:56:24 2005 From: al.floyd <@t> juno.com (al.floyd@juno.com) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Immunofluorescence problem-Non-specific binding appears in control slide Message-ID: <20050812.205624.2372.0.al.floyd@juno.com> Hello Nick, You may be running into the problem of Falck-Hillarp reactive cells, more commonly referred to as "SIF" cells. SIF stands for Small Intensely Fluorescent cells. This technique was quite popular back in the 1960's. In brief, any cell that contains epinephrine, nor-epi, or Dopamine will become very highly fluorescent after exposure to formalin. In the actual Falck-Hillarp technique, the exposure is to gaseous formaldehyde vapor, but you may be getting some of the same reaction with your procedure. Generally, Epi and Nor-epi fluoresce with a color that is very similar to flourescein, while Dopamine has a more orange color. There is actually a monograph available titled SIF cells. I forget the exact date, but think it is early '70's. I have a copy on my bookshelf, but at the moment am a long way from the bookshelf! Try a Google, Google Scholar or PubMed search for SIF cells - you should find quite a few publications. It was a very popular technique before the days of immunostains. Al Floyd Alton D. Floyd, Ph.D. 23126 South Shore Drive Edwardsburg, MI 49112 Cell: 574 215-0703 From lpwenk <@t> sbcglobal.net Sat Aug 13 00:01:11 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] HT Workforce In-Reply-To: Message-ID: Information available about the laboratory workforce in general: http://www.labmedicine.com/headlines/ascpnews/040205.html 72% of laboratory workers are older than the age of 40. Approximately 30% are older than the age of 50 Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Janice A Mahoney Sent: Thursday, August 11, 2005 4:49 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HT Workforce Hello, Does anyone know what the average age of registered Histo techs is? How many retire each year and how many new ones come into the field? Thanks for the help. Jan Omaha Janice Mahoney Histology/Cytology Coordinator Alegent Health Laboratory Phone (402)717-2889 Fax (402)717-5231 "We pledge to be creative, visionary leaders committed to holistic healthcare in the region" _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vanann702 <@t> skmc.gov.ae Sat Aug 13 02:35:49 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] VIP cleaning cycle Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E02F6D5CD@SKMCEMAIL.skmc.gov.ae> My Vip5 does just fine on a 7 day cleaning cycle. I use the shortened cleaning cycle for 6 days and do a long one once in the 7 day period. I am also fastidious about having a hot water flush once a month on the first 4 stations. My general solutions are 'rotated' every 7 days and the wax is as well - discard 1st, shift all up and renew last. Annieinarabia (formerly Annieinafrica) -----Original Message----- From: Fred Underwood [mailto:funderwood@mcohio.org] Sent: Thursday, August 11, 2005 8:10 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] VIP cleaning cycle Hi in Hisotland, This question is for those with a VIP 5 processor. Has anyone tried to get more than the recommended 5 runs from the cleaning xylene and alcohol? Thanks in adavance, Fred Underwood Mont. Co. Coroner Dayton, OH _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jennifer.l.hofecker <@t> Vanderbilt.Edu Sat Aug 13 07:53:20 2005 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] more about the gomori's on Frozen muscle Message-ID: <898D946569A27444B65667A49C07405207545D@mailbe06.mc.vanderbilt.edu> First, Thank you to those of you that responded. Especially those who understood that when doing modified Gomori's for a neuropathologist, muscle should stain green. I realize that in FFPE, muscle stains red, however our muscle has been staining correctly (green) until this week. I think that over the past several years, the neuropathologist might have noticed if we were staining the muscle the completely wrong color. (sorry, a little sarcastic) I am also not fixing the sections at all. We go right into hematoxylin. A private reply I received suggested that putting fresh slides in the freezer may be acting as "fixation" (lyophilization) thus, the fresh cuts aren't getting fixed. To those of you doing neuro muscle staining (remember, we're searching for normal to be GREEN) do you use fixation before staining? I would like to remind some of you that histonet is a tool for us to help each other out. It really is not for use as a weapon. I honestly needed advice, but I as usual, hesitated because I was afraid of the "less than kind" responses that seem to come with many of my posts. Thanks again to those who were kind enough to help me out. There is such a wealth of knowledge out there in histoland. Hopefully somebody will have the answer. Bracing for the next round, Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax From vanann702 <@t> skmc.gov.ae Sat Aug 13 08:06:44 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] more about the gomori's on Frozen muscle Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E02F6D5CF@SKMCEMAIL.skmc.gov.ae> Ran a neuropath lab for many years and did muscle bx's galore I used to cut my sections and store them at -20. the next day, id fix in methanol (I think for 5 mins), rinse in h2o, stain in harris' hx, another rinse, into the trichrome stain for 10-15 mins, quick rinse, few dips in weak acetic acid, DC+M Method from Carleton. Used it for many years with great success. FYI - this trichrome stain works well on FFPE - colours are a little different but it's a 'no hassle' stain. Works for me!! Hope this helps Annieinarabia -----Original Message----- From: Hofecker, Jennifer L [mailto:jennifer.l.hofecker@Vanderbilt.Edu] Sent: Saturday, August 13, 2005 4:53 PM To: histonet Subject: [Histonet] more about the gomori's on Frozen muscle First, Thank you to those of you that responded. Especially those who understood that when doing modified Gomori's for a neuropathologist, muscle should stain green. I realize that in FFPE, muscle stains red, however our muscle has been staining correctly (green) until this week. I think that over the past several years, the neuropathologist might have noticed if we were staining the muscle the completely wrong color. (sorry, a little sarcastic) I am also not fixing the sections at all. We go right into hematoxylin. A private reply I received suggested that putting fresh slides in the freezer may be acting as "fixation" (lyophilization) thus, the fresh cuts aren't getting fixed. To those of you doing neuro muscle staining (remember, we're searching for normal to be GREEN) do you use fixation before staining? I would like to remind some of you that histonet is a tool for us to help each other out. It really is not for use as a weapon. I honestly needed advice, but I as usual, hesitated because I was afraid of the "less than kind" responses that seem to come with many of my posts. Thanks again to those who were kind enough to help me out. There is such a wealth of knowledge out there in histoland. Hopefully somebody will have the answer. Bracing for the next round, Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcolclefa <@t> aol.com Sat Aug 13 16:14:55 2005 From: jcolclefa <@t> aol.com (John PJ Coleman) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Gomori Green Muscle trichrome. In-Reply-To: <200508131301.26542fe2750220@rly-yg01.mx.aol.com> References: <200508131301.26542fe2750220@rly-yg01.mx.aol.com> Message-ID: <211a15170008b530adf412b6223270a0@aol.com> Gomori "Green" muscle trichrome: I run a high volume IHC/Neuro/Muscle/Bone Marrow lab in VA and we had issues every once in a while with the red blotches appearing from Sigma's Gomori Trichrome solution. It sounds like witchcraft, but when the solution was agitated or mixed or dropped (oops) or freshly poured we noticed a greater likelihood of red artefactual staining. (nondescript blotchy red patches). We cut muscle frozen in liquid isopentane (n-methyl butane) cooled in liquid N2 at 4 microns, we do not fix at all, we do sometimes store slides at -70C before staining which does dessicate them, but we haven't linked this treatment to the spurious red blotches. try keeping your solution in a coplin jar, and don't agitate before staining. We pre counterstain in Mayer's Hamatoxylin from Biogenex from our IHC proc. for 3- 5 min. Rinse dh2o, gomori one step for 20 minutes, rinse tap h2o, aquamount. On Aug 13, 2005, at 1:01 PM, histonet-request@lists.utsouthwestern.edu wrote: > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. Re: Immunofluorescence problem-Non-specific binding appears > in control slide (al.floyd@juno.com) > 2. RE: HT Workforce (Lee & Peggy Wenk) > 3. RE: VIP cleaning cycle (Anne Van Binsbergen) > 4. more about the gomori's on Frozen muscle (Hofecker, Jennifer L) > 5. RE: more about the gomori's on Frozen muscle (Anne Van > Binsbergen) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Fri, 12 Aug 2005 20:56:24 -0400 > From: al.floyd@juno.com > Subject: Re: [Histonet] Immunofluorescence problem-Non-specific > binding appears in control slide > To: NSTYLOPOULOS@PARTNERS.ORG > Cc: histonet@lists.utsouthwestern.edu > Message-ID: <20050812.205624.2372.0.al.floyd@juno.com> > Content-Type: text/plain; charset=us-ascii > > Hello Nick, > > You may be running into the problem of Falck-Hillarp reactive cells, > more > commonly referred to as "SIF" cells. SIF stands for Small Intensely > Fluorescent cells. This technique was quite popular back in the > 1960's. > In brief, any cell that contains epinephrine, nor-epi, or Dopamine will > become very highly fluorescent after exposure to formalin. In the > actual > Falck-Hillarp technique, the exposure is to gaseous formaldehyde vapor, > but you may be getting some of the same reaction with your procedure. > Generally, Epi and Nor-epi fluoresce with a color that is very similar > to > flourescein, while Dopamine has a more orange color. > > There is actually a monograph available titled SIF cells. I forget the > exact date, but think it is early '70's. I have a copy on my > bookshelf, > but at the moment am a long way from the bookshelf! > > Try a Google, Google Scholar or PubMed search for SIF cells - you > should > find quite a few publications. It was a very popular technique before > the days of immunostains. > > Al Floyd > Alton D. Floyd, Ph.D. > 23126 South Shore Drive > Edwardsburg, MI 49112 > Cell: 574 215-0703 > > > > ------------------------------ > > Message: 2 > Date: Sat, 13 Aug 2005 01:01:11 -0400 > From: "Lee & Peggy Wenk" > Subject: RE: [Histonet] HT Workforce > To: "'Janice A Mahoney'" > Cc: 'Histonet submissions' > Message-ID: > > Content-Type: text/plain; charset="US-ASCII" > > Information available about the laboratory workforce in general: > > http://www.labmedicine.com/headlines/ascpnews/040205.html > 72% of laboratory workers are older than the age of 40. Approximately > 30% > are older than the age of 50 > > Peggy A. Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > > > Lee & Peggy Wenk > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Janice > A > Mahoney > Sent: Thursday, August 11, 2005 4:49 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] HT Workforce > > Hello, > Does anyone know what the average age of registered Histo techs is? > How > many retire each year and how many new ones come into the field? > Thanks for the help. > Jan > Omaha > > Janice Mahoney > Histology/Cytology Coordinator > Alegent Health Laboratory > Phone (402)717-2889 > Fax (402)717-5231 > > "We pledge to be creative, visionary leaders committed to holistic > healthcare in the region" > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 3 > Date: Sat, 13 Aug 2005 11:35:49 +0400 > From: "Anne Van Binsbergen" > Subject: RE: [Histonet] VIP cleaning cycle > To: "Fred Underwood" , > > Message-ID: > <0C44F1AAEE47D54DA4210A60AB206F5E02F6D5CD@SKMCEMAIL.skmc.gov.ae> > Content-Type: text/plain; charset="us-ascii" > > My Vip5 does just fine on a 7 day cleaning cycle. I use the shortened > cleaning cycle for 6 days and do a long one once in the 7 day period. I > am also fastidious about having a hot water flush once a month on the > first 4 stations. My general solutions are 'rotated' every 7 days and > the wax is as well - discard 1st, shift all up and renew last. > Annieinarabia (formerly Annieinafrica) > > -----Original Message----- > From: Fred Underwood [mailto:funderwood@mcohio.org] > Sent: Thursday, August 11, 2005 8:10 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] VIP cleaning cycle > > Hi in Hisotland, > > This question is for those with a VIP 5 processor. Has anyone tried to > get more than the recommended 5 runs from the cleaning xylene and > alcohol? > > Thanks in adavance, > > Fred Underwood > Mont. Co. Coroner > Dayton, OH > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 4 > Date: Sat, 13 Aug 2005 07:53:20 -0500 > From: "Hofecker, Jennifer L" > Subject: [Histonet] more about the gomori's on Frozen muscle > To: "histonet" > Message-ID: > <898D946569A27444B65667A49C07405207545D@mailbe06.mc.vanderbilt.edu> > Content-Type: text/plain; charset="iso-8859-1" > > First, > > Thank you to those of you that responded. Especially those who > understood that when doing modified Gomori's for a neuropathologist, > muscle should stain green. I realize that in FFPE, muscle stains red, > however our muscle has been staining correctly (green) until this > week. I think that over the past several years, the neuropathologist > might have noticed if we were staining the muscle the completely wrong > color. (sorry, a little sarcastic) > > I am also not fixing the sections at all. We go right into > hematoxylin. A private reply I received suggested that putting fresh > slides in the freezer may be acting as "fixation" (lyophilization) > thus, the fresh cuts aren't getting fixed. To those of you doing neuro > muscle staining (remember, we're searching for normal to be GREEN) do > you use fixation before staining? > > I would like to remind some of you that histonet is a tool for us to > help each other out. It really is not for use as a weapon. I honestly > needed advice, but I as usual, hesitated because I was afraid of the > "less than kind" responses that seem to come with many of my posts. > > Thanks again to those who were kind enough to help me out. There is > such a wealth of knowledge out there in histoland. Hopefully somebody > will have the answer. > > Bracing for the next round, > > Jennifer > > > > Jennifer Hofecker, HT (ASCP) > > Vanderbilt University Medical Center > > Division of Neuropathology > > (615) 343-0083 > > (615) 343-7089 fax > > > > ------------------------------ > > Message: 5 > Date: Sat, 13 Aug 2005 17:06:44 +0400 > From: "Anne Van Binsbergen" > Subject: RE: [Histonet] more about the gomori's on Frozen muscle > To: "Hofecker, Jennifer L" , > "histonet" > Message-ID: > <0C44F1AAEE47D54DA4210A60AB206F5E02F6D5CF@SKMCEMAIL.skmc.gov.ae> > Content-Type: text/plain; charset="us-ascii" > > Ran a neuropath lab for many years and did muscle bx's galore > I used to cut my sections and store them at -20. > the next day, id fix in methanol (I think for 5 mins), rinse in h2o, > stain in harris' hx, another rinse, into the trichrome stain for 10-15 > mins, quick rinse, few dips in weak acetic acid, DC+M > Method from Carleton. Used it for many years with great success. > FYI - this trichrome stain works well on FFPE - colours are a little > different but it's a 'no hassle' stain. > Works for me!! > Hope this helps > Annieinarabia > > -----Original Message----- > From: Hofecker, Jennifer L [mailto:jennifer.l.hofecker@Vanderbilt.Edu] > Sent: Saturday, August 13, 2005 4:53 PM > To: histonet > Subject: [Histonet] more about the gomori's on Frozen muscle > > First, > > Thank you to those of you that responded. Especially those who > understood that when doing modified Gomori's for a neuropathologist, > muscle should stain green. I realize that in FFPE, muscle stains red, > however our muscle has been staining correctly (green) until this week. > I think that over the past several years, the neuropathologist might > have noticed if we were staining the muscle the completely wrong color. > (sorry, a little sarcastic) > > I am also not fixing the sections at all. We go right into hematoxylin. > A private reply I received suggested that putting fresh slides in the > freezer may be acting as "fixation" (lyophilization) thus, the fresh > cuts aren't getting fixed. To those of you doing neuro muscle staining > (remember, we're searching for normal to be GREEN) do you use fixation > before staining? > > I would like to remind some of you that histonet is a tool for us to > help each other out. It really is not for use as a weapon. I honestly > needed advice, but I as usual, hesitated because I was afraid of the > "less than kind" responses that seem to come with many of my posts. > > Thanks again to those who were kind enough to help me out. There is > such > a wealth of knowledge out there in histoland. Hopefully somebody will > have the answer. > > Bracing for the next round, > > Jennifer > > > > Jennifer Hofecker, HT (ASCP) > > Vanderbilt University Medical Center > > Division of Neuropathology > > (615) 343-0083 > > (615) 343-7089 fax > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 21, Issue 17 > **************************************** > JPJC-757 335-2159 http://journals.aol.com/jcolclefa/Waytoomuchboringcraptosayusually/ From jkiernan <@t> uwo.ca Sat Aug 13 23:23:13 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Immunofluorescence problem-Non-specific binding appears incontrol slide References: <4063C6544431714FA0EDA3D9138CB7A4A81194@PHSXMB5.partners.org> Message-ID: <42FEC731.57251F10@uwo.ca> Al Floyd's astute suggestion of a Falck-Hillarp reaction is certainly a possible cause of "specific-looking" autofluorescence. In rat intestine and its associated connective tissue you can expect a F-H reaction in mast cells and classical argentaffin cells, from their high content of serotonin. (The noradrenaline in sympathetic axons doesn't give a localized fluorescent product with ordinary formaldehyde solutions.) If your unwanted fluorescence is definitely due to binding of the FITC-labelled secondary antiserum, a possible cause may be binding of the FITC-secondary to something in your "blocking" protein: You wrote: > ... Block with 10% donkey serum (I tried adding 10% > rat serum or 3% BSA) for 2hrs (I tried also blocking > overnight... or I added Background buster (from > Inovvex) and Fc receptor blocker ... Not all your "blocking" substances are identified, but they are all intended to bind to the tissue. Rat serum may not be good stuff to put on a rat tissue if the secondary antiserum is an anti-(rat immunoglobulin). For competitive blocking of low affinity binding of primary and labelled antibodies it's probably best to go with either a "generic protein" such as bovine albumin or a non-immune serum from the host species of the secondary. Before using any trade-name product, find out what it is and how it may work. Follow up aome of the references in the small print at the bottom of the brochure. Clear thinking (what sticks to what, and why?) is the paramount principle in immunohistochemistry. John Kiernan Anatomy, UWO London, Canada. ______________________________ "Stylopoulos, Nicholas" wrote: > > Hello everybody and thank you for taking the time to read this message. I have > been struggling with immunofluorescence and I have the following problem: My > control slide (secondary antibody only) appears to have specific staining. I > have repeated the assay many times (Also, I showed my control slide to many > people and they asked me whether I am sure that I have not added my primary > antibody as well!). My primary antibody is goat anti-rat (against chromogranin > A). I have also tried rabbit anti-rat. My secondary is donkey anti-goat FITC > conjugated (I have tried donkey anti-rabbit and goat anti-rabbit). Basically, I > am trying to visualize the endocrine cells in the gastrointestinal tract. This > is my protocol: > > Tissue: rat ileum-snap frozen in isopentane > > Sections: Frozen sections 3-5?m > > Fixation: 4% paraformaldehyde (I tried also acetone -20C from 30secs to 20min > and it seems that acetone destroys the histology of my sections) > > Washx3 with filtered PBS 1X > > Triton x-100 0.05% for 5min > > Wash x1 with filtered PBS 1X > > Block with 10% donkey serum (I tried adding 10% rat serum or 3% BSA) for 2hrs (I > tried also blocking overnight... or I added Background buster (from Inovvex) and > Fc receptor blocker (from Innovex too) or sodium azide 0.1%) > > Primary Antibody in PBS for 1hr > > Washx3 with filtered PBS 1X > > Secondary antibody in PBS for 1 hr > > Washx6 with filtered PBS 1X > > Mount with DAPI > > > > Any suggestions will be extremely appreciated....Thanks again! > > > > Nick > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NSTYLOPOULOS <@t> PARTNERS.ORG Sun Aug 14 11:31:24 2005 From: NSTYLOPOULOS <@t> PARTNERS.ORG (Stylopoulos, Nicholas) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Immunofluorescence problem-Non-specific binding appears in control slide Message-ID: <4063C6544431714FA0EDA3D9138CB7A4A81195@PHSXMB5.partners.org> Thank you so much for all your suggestions! I found the papers on Falck-Hillarp reaction that Al [Floyd] suggested. It sounds that it may be possible to be the cause of the specific-looking autofluorescence, because my specific binding occurs (mostly) in the lamina propria of almost each single villus and the outer layers of my specimen (in the connective tissue and the muscular layer). This is very similar to what John [Kiernan] suggested in his response. This "specific binding" is not related to the nature of the secondary antibody (FITC, TRITC, anti-goat or anti-rabbit) and the concentration (I tried dilutions up to 1:5000). I can even see very nice "rings" around the nuclei (like something is stained in the ER-Golgi). Also it is very similar to the picture I get when I add my primary (which is anti-chromogranin A). It looks like almost the same cells are stained! As a matter of fact, I thought that I had cross-contamination, so I started to use filtered tip pipettes and prepare new reagents every time I would do the assay. The only two arguments against the Falck-Hillarp reaction are that 1) the fluorescence is prominent only after I add my secondary antibody and 2) I tried fixing the tissues with Acetone and I got a slightly better picture but not extremely different. However, I did not try acetone many times because it would destroy the histology and additionally the DAPI staining was kind of diffused and not very helpful (which I assume is expected when acetone is used as a fixative). Finally, to answer Julia's [Edgar] question, I did try an anti-rabbit secondary which did not give me a different picture. In the next few days, I would like to rule out the Falck-Hillarp issue and it would be amazing if you could give me some info on the following two questions: 1) I snap-freeze the tissues in isopentane. Is it possible that isopentane might create the problem? The reason I am asking is that -from what I read- in the initial description of Falck-Hillarp method, the tissues were snap-frozen in propane. 2) What fixation method would you suggest instead of paraformaldehyde? Is is possible to use Acetone but improve the histologic picture? Are there any other methods that you would suggest? I have already harvested the tissues after I snap-frozen them in isopentane (and some in liquid nitrogen). Thank you again for your help!!! Nick Nicholas Do you get the same problem with the anti-rabbit antibodies? Anit- goat can be very 'dirty' I find. Julia Julia Edgar BSc (Hons), PhD Applied Neurobiology Group University of Glasgow Veterinary School Bearsden Road Glasgow G61 1QH Tel: 0141 330 5818 e-mail: je22r@udcf.gla.ac.uk Al Floyd's astute suggestion of a Falck-Hillarp reaction is certainly a possible cause of "specific-looking" autofluorescence. In rat intestine and its associated connective tissue you can expect a F-H reaction in mast cells and classical argentaffin cells, from their high content of serotonin. (The noradrenaline in sympathetic axons doesn't give a localized fluorescent product with ordinary formaldehyde solutions.) If your unwanted fluorescence is definitely due to binding of the FITC-labelled secondary antiserum, a possible cause may be binding of the FITC-secondary to something in your "blocking" protein: You wrote: > ... Block with 10% donkey serum (I tried adding 10% > rat serum or 3% BSA) for 2hrs (I tried also blocking > overnight... or I added Background buster (from > Inovvex) and Fc receptor blocker ... Not all your "blocking" substances are identified, but they are all intended to bind to the tissue. Rat serum may not be good stuff to put on a rat tissue if the secondary antiserum is an anti-(rat immunoglobulin). For competitive blocking of low affinity binding of primary and labelled antibodies it's probably best to go with either a "generic protein" such as bovine albumin or a non-immune serum from the host species of the secondary. Before using any trade-name product, find out what it is and how it may work. Follow up aome of the references in the small print at the bottom of the brochure. Clear thinking (what sticks to what, and why?) is the paramount principle in immunohistochemistry. John Kiernan Anatomy, UWO London, Canada. Hello Nick, You may be running into the problem of Falck-Hillarp reactive cells, more commonly referred to as "SIF" cells. SIF stands for Small Intensely Fluorescent cells. This technique was quite popular back in the 1960's. In brief, any cell that contains epinephrine, nor-epi, or Dopamine will become very highly fluorescent after exposure to formalin. In the actual Falck-Hillarp technique, the exposure is to gaseous formaldehyde vapor, but you may be getting some of the same reaction with your procedure. Generally, Epi and Nor-epi fluoresce with a color that is very similar to flourescein, while Dopamine has a more orange color. There is actually a monograph available titled SIF cells. I forget the exact date, but think it is early '70's. I have a copy on my bookshelf, but at the moment am a long way from the bookshelf! Try a Google, Google Scholar or PubMed search for SIF cells - you should find quite a few publications. It was a very popular technique before the days of immunostains. Al Floyd Alton D. Floyd, Ph.D. 23126 South Shore Drive Edwardsburg, MI 49112 Cell: 574 215-0703 From mami_73 <@t> yahoo.com Sun Aug 14 13:58:16 2005 From: mami_73 <@t> yahoo.com (adrienn kiss) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] mouse heart Message-ID: <20050814185816.68933.qmail@web54101.mail.yahoo.com> Hi, I am new on the list and also in mouse heart histology. I am wondering what is the best way to take a heart for good histology. Until in situ, how long the heart can be left without fixation before any signs of necrosis would appear by histology? I mean in a dead animal how long it can be left cyanotic before major effects on morphology of the myofibers, cell size, or immunohistochemistry to detect apoptosis, etc. I would normally take the heart asap, however, it is not possible always. Would perfusion with fixative always be suggested? Is perfusion of the aorta with fixative also suggested? Would retrograde perfusion damage the enothelium? Any help is appreciated, Thanks a lot, Adrienn Kis, Postdoctoral research fellow univeristy of Cambridge, England, ____________________________________________________ Start your day with Yahoo! - make it your home page http://www.yahoo.com/r/hs From nyilmaz <@t> mersin.edu.tr Mon Aug 15 01:33:48 2005 From: nyilmaz <@t> mersin.edu.tr (nyilmaz@mersin.edu.tr) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Nikon relay lens Message-ID: <20050815063348.6D9AA45587@mail.mersin.edu.tr> Hi All... We're using an Olympus BX 50 light microscope attached with Nikon Coolpix 5000 digital camera. I'm wondering what the mangnification value of MDC relay lens between the camera and the microscope is. I could'nt find any information from internet or other sources. We can't estimate the final magnification value of pictures correctly without knowing that. Thanks for the help. Dr. Necat YILMAZ Mersin University Medical School Histology & Embryology Dep. Mersin/TURKEY From Heather.A.Harper <@t> pcola.med.navy.mil Mon Aug 15 06:48:36 2005 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] CAP Info Needed Message-ID: <807FE48C5A7CC940B973B58D32E701431A850CCC@nhpens-exch1.pcola.med.navy.mil> Our hospital did the histology CAP, where you send in the requested slides, and you can earn CE points by doing the final critique. I want and need to know, when they send you back your results, is there any forms you have to fill out like what corrective action was taken etc.. Since this is something new that CAP has started, we are not sure what our next move is and I have already received the next CAP packet and must have slides submitted no later than Sept. 2 Also to get credit for the Critique, do I need to create a password and log onto the CAP website? Any info is very much appreciated. Heather A. Harper Histology Supervisor Naval Hospital of Pensacola, FL From ktalbot <@t> jhmi.edu Mon Aug 15 07:30:41 2005 From: ktalbot <@t> jhmi.edu (Karen Fox-Talbot) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] immuno peroxidase and autoradiography Message-ID: I would like to do a combination stain of immunperoxidase and autoradiography (using Indium). Does anyone have any experience with this? Karen Fox-Talbot Johns Hopkins University Department of Pathology 410-614-0915 From Charlotte.Kopczynski <@t> baycare.org Mon Aug 15 09:14:35 2005 From: Charlotte.Kopczynski <@t> baycare.org (Kopczynski, Charlotte) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Breast Implant Storage Message-ID: <5452D66669CFC540B0452115ED26AF1F0191AEFC@bcexch02.bcad.baycare.org> I'm interested in how long you are keeping breast implants that have been removed. I have seen the recommendations on the CAP site but they do not address the length of time these specimens should be kept. I am trying to get my Risk management Department to make a decision. Thanks, Charlotte Kopczynski,HTL(ASCP) Baycare Pathology Manager Phone: 727-461-8246 Fax: 727-462-7597 Confidential: This electronic message and all contents contain information from BayCare Health System which may be privileged, confidential or otherwise protected from disclosure. The information is intended to be for the addressee only. If you are not the addressee, any disclosure, copy, distribution or use of the contents of this message is prohibited. If you have received this electronic message in error, please notify the sender and destroy the original message and all copies. From dsv22 <@t> hotmail.com Mon Aug 15 09:56:21 2005 From: dsv22 <@t> hotmail.com (Denise Van Eaton) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Shandon Citadel 2000 for sale Message-ID: We have 3, due to some reinstrumentation. Ages are 1983, 1993,and 1996. All three are the 2000 model, no vacuum.Two of the three are in good working order, the third is missing a paraffin bath (cracked). Let us know, we'd sure like to find them all a new home! thanks,denise _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From laurie.colbert <@t> huntingtonhospital.com Mon Aug 15 10:00:24 2005 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Breast Implant Storage Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628005653744@EXCHANGE1.huntingtonhospital.com> Six months - the same as hardware. We used to keep them indefinitely, but you can imagine what a mess that was. Laurie Colbert -----Original Message----- From: Kopczynski, Charlotte [mailto:Charlotte.Kopczynski@baycare.org] Sent: Monday, August 15, 2005 7:15 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Breast Implant Storage I'm interested in how long you are keeping breast implants that have been removed. I have seen the recommendations on the CAP site but they do not address the length of time these specimens should be kept. I am trying to get my Risk management Department to make a decision. Thanks, Charlotte Kopczynski,HTL(ASCP) Baycare Pathology Manager Phone: 727-461-8246 Fax: 727-462-7597 Confidential: This electronic message and all contents contain information from BayCare Health System which may be privileged, confidential or otherwise protected from disclosure. The information is intended to be for the addressee only. If you are not the addressee, any disclosure, copy, distribution or use of the contents of this message is prohibited. If you have received this electronic message in error, please notify the sender and destroy the original message and all copies. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RCHIOVETTI <@t> aol.com Mon Aug 15 10:38:55 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Nikon relay lens Message-ID: <1c3.2eb8ab83.3032110f@aol.com> In a message dated 8/14/2005 11:35:35 PM US Mountain Standard Time, nyilmaz@mersin.edu.tr writes: > We're using an Olympus BX 50 light microscope attached with Nikon Coolpix > 5000 digital camera. I'm wondering what the mangnification value of MDC relay > lens between the camera and the microscope is. Dr. Yilmaz, I recently set up a system on a Leica microscope, using a Nikon Coolpix 8800. We did the following: 1. We obtained a stage micrometer, which is a precise ruling that is etched on a microscope slide. The one we used was 1 mm overall length, with smallest rulings of 0.010 mm (10 micrometers). 2. We did not use the MDC relay lens, but we purchased an adapter from another company. 3. On our system, we had to use the optical zoom of the camera to obtain a full field of view (eliminate the vignetting, or "clipping" of the image at the corners). 4. Since the zoom on the camera is infinitely adjustable, we had to decide on a reproducible zoom setting that always gave the same magnification. For the Coolpix 8800 and our adapter, we opted to zoom to the maximum setting. It's 10x on the 8800; I'm not sure what it is on the 5000. 5. To make measurements we set the camera to this maximum (10x) zoom, and took pictures of the stage micrometer with each objective. From there, it's relatively easy to measure the spacing of the 10 micrometer lines and to calculate a total magnification. This was the only way we could come up with to make measurements, since the camera's zoom is so variable. It's a small inconvenience, but the camera works great with this set-up. You could probably use a similar procedure, provided you are using the camera's optical zoom to fill the field of view with the MDC relay lens. The trick is to use a zoom setting that is reproducible. Hope this is of some help. Best regards, Robert (Bob) Chiovetti, Ph.D. The Microscope Works Arizona's Microscopy Resource 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From histology <@t> gradymem.org Mon Aug 15 10:50:19 2005 From: histology <@t> gradymem.org (histology@gradymem.org) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] handling fee specimens/tests sent to ref. lab Message-ID: <2b50526ed7.26ed72b505@onenet.net> Does anyone charge the patient a handling fee for specimens sent to a reference lab? We send our cytologies out and used to charge a handling fee. Any comments? Angie From gcallis <@t> montana.edu Mon Aug 15 13:46:24 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] fixation for beta galactosidase staining, whole mouse embryo Message-ID: <6.0.0.22.1.20050815124354.01b18b88@gemini.msu.montana.edu> You need to use a paraformaldehyde fixative that contains EGTA that works best with Beta Gal staining, if you contact me, I will send you the information and the Beta Gal bible on how to deal with staining for this, very handy and informative. Whole mount staining is not difficult, and very elegant. 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 gsmith <@t> confocal.com Mon Aug 15 13:48:17 2005 From: gsmith <@t> confocal.com (Glenn Smith) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Flourescent labels Message-ID: <005801c5a1c9$eb1afcc0$1f05a8c0@confocal.com> Patsy, I know one of our customers use our TISSUEscope instrument to scan immunofluorescence and immunohistochemistry (RGB) slides and had imaged fluorescence ffpe tissues. When I asked her about your question, her comments to me regarding the TISSUEscope, were that formalin fixed paraffin embedded is no different from the frozen sections. Our systems employ the narrow bandwidth filters that Tim Morken speaks of below and are laser-based. The specific system cited above uses blue (488nm), green (532nm) and red (635nm) laser sources (with relevant filters) to digitally capture the entire sample in either brightfield (rgb) or fluoresence contrast modes. We have other customer labs that have different laser combinations depending on the fluorophores of interest and have supplied a variety of sources between 400 - 800 nm (violet, blue, green, yellow, red, ruby) matched up with the appropriate filter combinations. Glenn Glenn Smith, P.Eng. ph: 519.886.9013 x38 mobile: 519.498.0614 email: gsmith@confocal.com Biomedical Photometrics Inc./GeneFocus A12-550 Parkside Dr. Waterloo, ON N2L 5V4 Widefield Laser Scanning Instruments and Software for Fluorescence and Brightfield Imaging www.confocal.com The information in this e-mail message, including any attachments, is confidential. It is intended only for the addressee and any use or disclosure by others is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete all copies of the message. Thank you. Message: 19 Date: Fri, 12 Aug 2005 16:31:52 -0400 From: "Morken, Tim - Labvision" Subject: RE: [Histonet] Flourescent labels To: 'Patsy Ruegg' , 'Histonet' Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D002@usca0082k08.labvision.apogent.com> Content-Type: text/plain Patsy, The average routine fluorescent system uses "longpass" emission filters that let almost all the light from blue to red through. Therefore you will see many colors of autofluoresce. Since the autofluorescence is usually a different color than your target fluorchrome, the way you can avoid autofluoresence is to find filters that block the autofluorescent wavelength. It takes more effort to coordinate the different wavelengths associated with the fluorochrome excitation and emission vs the autofluorescent excitation and emission, but by using narrow-bandwidth filters (20-40 nm bandwidth) it is usually possible. Both Omega and Chroma have these filters. 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 Patsy Ruegg Sent: Friday, August 12, 2005 1:18 PM To: 'Histonet' Subject: [Histonet] Flourescent labels Folks, I was under the general impression that most flourescent labels were done on frozen tissue because of problems with auto flourescence associated with aldehyde fixation and paraffin processing. Has the technology developed so that these new flourescent labels such as the Alexa dyes can now be used on ffpe tissues? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From joel <@t> srodes.com Mon Aug 15 14:40:16 2005 From: joel <@t> srodes.com (Joel) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Air bubbles Message-ID: <000601c5a1d1$2b636c20$0400a8c0@joel> I am preparing very nice slides of frozen tissue specimens in the course of performing Mohs Micrographic surgery. However, days later air bubbles form and the tissue begins to deterioate and become unrecognizable. Is any one else having this problem, and do you have a solution? From kccatunda <@t> terra.com.br Mon Aug 15 16:13:24 2005 From: kccatunda <@t> terra.com.br (kccatunda) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Headache X pregnancy Message-ID: Does anybody has complains of preagnant technicians with headaches because of the smell of reagents such as xylol? What are the precautions you have with preagnancy? Our mounting and staining are mannual procedures but we do have one exaustor for fumes that covers the staining battery. My worry is that we don?t use coverslippers, instead we use a kind of automotive coating (yeah... I know... bad bad bad...). Does somebody has any idea to substitute it without using coverslippers? Thanks again Katia From vd38 <@t> georgetown.edu Mon Aug 15 16:51:55 2005 From: vd38 <@t> georgetown.edu (Vernon Dailey) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] HDAC Primary Antibodies Message-ID: <43010E7B.3060705@georgetown.edu> Hi Histonetters, I have just been handed a project in which I need to run IHC for several HDAC proteins. I will be staining FFPE human tissue. Many of the antibodies out there are not characterized for IHC. I was hoping that some of you may have had some positive experience with HDAC 1, 2, 3, 4, 5, 6, 7, or 8 for IHC or IF. If you have had some success please fill me in on the vendor and clone. Thanks in advance for all of your help. -Vernon From jennifer.l.hofecker <@t> Vanderbilt.Edu Mon Aug 15 21:15:31 2005 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Headache X pregnancy References: Message-ID: <898D946569A27444B65667A49C074052075462@mailbe06.mc.vanderbilt.edu> Hi Katia, When I was pregnant, I was extremely sensitive to xylene. It was like I had a bionic nose or something. I found that if I avoided tasks like changing processors and coverslipping for long periods of time, I was fine. My supervisor was very good about putting me on rotations where I didn't have much xylene contact. This seems like the easiest solution, eventually as you know, your tech won't be pregnant anymore and will most likely lose her bionic powers. Just thought I'd let you know you aren't alone. Have a good week, Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of kccatunda Sent: Mon 8/15/2005 4:13 PM To: histonet Subject: [Histonet] Headache X pregnancy Does anybody has complains of preagnant technicians with headaches because of the smell of reagents such as xylol? What are the precautions you have with preagnancy? Our mounting and staining are mannual procedures but we do have one exaustor for fumes that covers the staining battery. My worry is that we don?t use coverslippers, instead we use a kind of automotive coating (yeah... I know... bad bad bad...). Does somebody has any idea to substitute it without using coverslippers? Thanks again Katia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From stevenk <@t> med.usyd.edu.au Mon Aug 15 23:54:23 2005 From: stevenk <@t> med.usyd.edu.au (stevenk) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] GFAP Immunohistochemistry on frozen tissue. Message-ID: I was wondering if anyone can give me any protocols on GFAP immunohistochemistry (IHC) on frozen tissue. Because we wish to preserve the DNA, we would like to avoid paraformaldehyde fixation and microwave antigen retrieval. Thanks Stephen -- Stephen Kum Jew Senior Technical Officer Neuropathology Department of Pathology Blackburn Building D06 University of Sydney NSW 2006 Australia Ph: 61 2 9351 6143 Fax: 61 2 9351 3429 From jhr1x <@t> clinmed.gla.ac.uk Tue Aug 16 02:28:36 2005 From: jhr1x <@t> clinmed.gla.ac.uk (Mr James Hugh Reilly) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Endogenous Peroxidase Message-ID: <1124177316.4c92479cjhr1x@clinmed.gla.ac.uk> I have been asked to try a new Immumohistochemistry (IHC) technique which uses 1% hydrogen peroxide and 2% sodium nitride in phosphate buffer solution. Does anyone know what sodium nitride Is? and where can it be purchased? Jim Reilly University Dept. Medicine Level 3 QEB Royal Infirmary 10 Alexandra Parade Glasgow G31 2ER From sulekhababy <@t> yahoo.co.uk Tue Aug 16 03:59:14 2005 From: sulekhababy <@t> yahoo.co.uk (Sulekha Ravi) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Grinding & polishing of resin sections. Message-ID: <20050816085914.98363.qmail@web25001.mail.ukl.yahoo.com> Dear Histonetters Please help me in overcoming this problem. We cut PMMA embedded tissue sections using Isomet. These sections have to be ground and polished using 'Ecomet grinder polisher'. After grinding & polishing using grinding papers and diamond paste of different sizes we find the sections contain lots of debris.What can I do to clean these resin sections before it is stained? Thanks Sulekha --------------------------------- To help you stay safe and secure online, we've developed the all new Yahoo! Security Centre. From PMonfils <@t> Lifespan.org Tue Aug 16 08:41:57 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Endogenous Peroxidase Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175AE@lsexch.lsmaster.lifespan.org> Sodium nitride is Na3N, but I don't think you will be able to purchase it anywhere. It is a compound that was long believed to be chemically impossible, but which only recently was experimentally synthesized for the first time by deposition of atomic sodium and nitrogen beams onto a supercooled sapphire in a vacuum. I doubt that the compound is commercially available. Perhaps it is sodium nitrite that is called for in your protocol? Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Mr > James Hugh Reilly > Sent: Tuesday, August 16, 2005 12:28 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Endogenous Peroxidase > > I have been asked to try a new Immumohistochemistry (IHC) technique which > uses 1% hydrogen peroxide and 2% sodium nitride in phosphate buffer > solution. Does anyone know what sodium nitride Is? and where can it be > purchased? > > Jim Reilly > > University Dept. Medicine > Level 3 QEB > Royal Infirmary > 10 Alexandra Parade > Glasgow > G31 2ER > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jkiernan <@t> uwo.ca Tue Aug 16 08:51:48 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Endogenous Peroxidase References: <1124177316.4c92479cjhr1x@clinmed.gla.ac.uk> Message-ID: <4301EF74.DCDF1755@uwo.ca> I think there must be a mistake in the name. Sodium nitride (Na3N) is a grey solid formed by heating metallic sodium in nitrogen. It is instantly decomposed by water, so a solution in phosphate buffer could not be made. -- ------------------------------- 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 _______________________________ Mr James Hugh Reilly wrote: > > I have been asked to try a new Immumohistochemistry (IHC) technique which uses 1% hydrogen peroxide and 2% sodium nitride in phosphate buffer solution. Does anyone know what sodium nitride Is? and where can it be purchased? > > Jim Reilly > > University Dept. Medicine > Level 3 QEB > Royal Infirmary > 10 Alexandra Parade > Glasgow > G31 2ER > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Tue Aug 16 08:52:14 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Endogenous Peroxidase Message-ID: <5D2189E74151CC42BEC02906BA8996322B90DD@exchsrv01.barrynet.barry.edu> Sodium nitride, Na3N, is a rather unstable compound first prepared in 2002. The synthesis is notoriously difficult. I do not think it is commercially available. I think that the "nitride" in your recipe is a typographical error in "nitrite". Sodium nitrite is available from both Aldrich and Fisher. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mr James Hugh Reilly Sent: Tuesday, August 16, 2005 3:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Endogenous Peroxidase I have been asked to try a new Immumohistochemistry (IHC) technique which uses 1% hydrogen peroxide and 2% sodium nitride in phosphate buffer solution. Does anyone know what sodium nitride Is? and where can it be purchased? Jim Reilly University Dept. Medicine Level 3 QEB Royal Infirmary 10 Alexandra Parade Glasgow G31 2ER _______________________________________________ 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 bwhitaker <@t> brownpathology.com Tue Aug 16 09:39:24 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Breast Implant Storage In-Reply-To: <5452D66669CFC540B0452115ED26AF1F0191AEFC@bcexch02.bcad.baycare.org> Message-ID: <002701c5a270$4bdfc440$3601a8c0@brownpathology.net> I would be interested in hearing what replies you get off-line. I don't have a written policy for them, but am currently keeping them for 8-12 months. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kopczynski, Charlotte Sent: Monday, August 15, 2005 9:15 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Breast Implant Storage I'm interested in how long you are keeping breast implants that have been removed. I have seen the recommendations on the CAP site but they do not address the length of time these specimens should be kept. I am trying to get my Risk management Department to make a decision. Thanks, Charlotte Kopczynski,HTL(ASCP) Baycare Pathology Manager Phone: 727-461-8246 Fax: 727-462-7597 Confidential: This electronic message and all contents contain information from BayCare Health System which may be privileged, confidential or otherwise protected from disclosure. The information is intended to be for the addressee only. If you are not the addressee, any disclosure, copy, distribution or use of the contents of this message is prohibited. If you have received this electronic message in error, please notify the sender and destroy the original message and all copies. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfavara <@t> niaid.nih.gov Tue Aug 16 09:42:31 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Myelin Basic Protein Message-ID: I have had success with Santa Cruz MBP[C-16] sc13914 and Serotec Rat Anti-MBP[36-50] Clone 14 on FFPE mouse tissue! 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: Thomas Crowell [mailto:Thomas.Crowell@biogenidec.com] Sent: Friday, August 12, 2005 11:21 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Myelin Basic Protein Is anyone using an antibody to MBP on FFPE mouse tissues? Thanks Tom Crowell BiogenIdec Cambridge, MA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lao_ji <@t> yahoo.com Tue Aug 16 10:35:49 2005 From: lao_ji <@t> yahoo.com (Jimmy Lao) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Problem in ISH on section Message-ID: <20050816153549.59777.qmail@web30712.mail.mud.yahoo.com> Dear Histoneters, My friend recently meets a problem in ISH on section, he still had good result two months ago. Here is the problem: Just during past a month, he couldn't get BM purple staining on the ISH, the protocol is on the web: http://saturn.med.nyu.edu/research/dg/joynerlab/protocols.html He said nornally BM purple overnight get the result, now it got a week. He already change solution, retry, it still not works. Any people met this slowly staining problem, and overcome it. Pls tell me the resolution. Thanks! Jimmy ____________________________________________________ Start your day with Yahoo! - make it your home page http://www.yahoo.com/r/hs From RCHIOVETTI <@t> aol.com Tue Aug 16 11:03:48 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Grinding & polishing of resin sections. Message-ID: <194.4500e299.30336864@aol.com> In a message dated 8/16/2005 2:00:20 AM US Mountain Standard Time, sulekhababy@yahoo.co.uk writes: > After grinding &polishing using grinding papers and diamond paste of > different sizes we find the sections contain lots of debris.What can I do to clean > these resin sections before it is stained? > Sulekha, Can you completely remove the plastic from the sections before staining? This would certainly remove the bulk of the contaminants, since they're getting embedded in the sections during the polishing process. It would be similar to the deparaffinization of paraffin sections. You could probably use 2-methoxy ethyl acetate to remove the methacrylate plastic. Then you would simply have the specimens remaining on the slide, with no plastic Wash and rehydrate thoroughly after the "deacrylation," and proceed with staining. See the following link: < http://www.ecmjournal.org/journal/supplements/vol007supp01/pdf/vol007supp01a82.pdf>. Best, Bob Robert (Bob) Chiovetti, Ph.D. The Microscope Works Arizona's Microscopy Resource 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From tmmrosla <@t> healtheast.org Tue Aug 16 11:05:34 2005 From: tmmrosla <@t> healtheast.org (Mrosla, Tina M) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] RE: Handling fee specimens Message-ID: <22E6C4C32AFB3E44B7056C7EC3E1F204DC46FD@HECLUSTER.HEALTHEAST.LOC> We charge $25 for a handling fee for all specimens that need to be sent to a reference lab for additional testing. The only thing that we do not charge for is consults requested by our pathologists. Second opinions requested by the physician or patient do, however. Tina Mrosla St. Joseph's Hospital St. Paul, MN -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Monday, August 15, 2005 12:27 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 21, Issue 19 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. mouse heart (adrienn kiss) 2. Nikon relay lens (nyilmaz@mersin.edu.tr) 3. CAP Info Needed (Heather.A.Harper@pcola.med.navy.mil) 4. immuno peroxidase and autoradiography (Karen Fox-Talbot) 5. Breast Implant Storage (Kopczynski, Charlotte) 6. Shandon Citadel 2000 for sale (Denise Van Eaton) 7. RE: Breast Implant Storage (Laurie Colbert) 8. Re: Nikon relay lens (RCHIOVETTI@aol.com) 9. handling fee specimens/tests sent to ref. lab (histology@gradymem.org) ---------------------------------------------------------------------- Message: 1 Date: Sun, 14 Aug 2005 11:58:16 -0700 (PDT) From: adrienn kiss Subject: [Histonet] mouse heart To: histonet@lists.utsouthwestern.edu Message-ID: <20050814185816.68933.qmail@web54101.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi, I am new on the list and also in mouse heart histology. I am wondering what is the best way to take a heart for good histology. Until in situ, how long the heart can be left without fixation before any signs of necrosis would appear by histology? I mean in a dead animal how long it can be left cyanotic before major effects on morphology of the myofibers, cell size, or immunohistochemistry to detect apoptosis, etc. I would normally take the heart asap, however, it is not possible always. Would perfusion with fixative always be suggested? Is perfusion of the aorta with fixative also suggested? Would retrograde perfusion damage the enothelium? Any help is appreciated, Thanks a lot, Adrienn Kis, Postdoctoral research fellow univeristy of Cambridge, England, ____________________________________________________ Start your day with Yahoo! - make it your home page http://www.yahoo.com/r/hs ------------------------------ Message: 2 Date: Mon, 15 Aug 2005 09:33:48 +0300 From: Subject: [Histonet] Nikon relay lens To: Message-ID: <20050815063348.6D9AA45587@mail.mersin.edu.tr> Content-Type: text/plain; charset="ISO-8859-9" Hi All... We're using an Olympus BX 50 light microscope attached with Nikon Coolpix 5000 digital camera. I'm wondering what the mangnification value of MDC relay lens between the camera and the microscope is. I could'nt find any information from internet or other sources. We can't estimate the final magnification value of pictures correctly without knowing that. Thanks for the help. Dr. Necat YILMAZ Mersin University Medical School Histology & Embryology Dep. Mersin/TURKEY ------------------------------ Message: 3 Date: Mon, 15 Aug 2005 06:48:36 -0500 From: Heather.A.Harper@pcola.med.navy.mil Subject: [Histonet] CAP Info Needed To: histonet@lists.utsouthwestern.edu Message-ID: <807FE48C5A7CC940B973B58D32E701431A850CCC@nhpens-exch1.pcola.med.navy.mil> Content-Type: text/plain Our hospital did the histology CAP, where you send in the requested slides, and you can earn CE points by doing the final critique. I want and need to know, when they send you back your results, is there any forms you have to fill out like what corrective action was taken etc.. Since this is something new that CAP has started, we are not sure what our next move is and I have already received the next CAP packet and must have slides submitted no later than Sept. 2 Also to get credit for the Critique, do I need to create a password and log onto the CAP website? Any info is very much appreciated. Heather A. Harper Histology Supervisor Naval Hospital of Pensacola, FL ------------------------------ Message: 4 Date: Mon, 15 Aug 2005 08:30:41 -0400 From: "Karen Fox-Talbot" Subject: [Histonet] immuno peroxidase and autoradiography To: Message-ID: Content-Type: text/plain; charset=US-ASCII I would like to do a combination stain of immunperoxidase and autoradiography (using Indium). Does anyone have any experience with this? Karen Fox-Talbot Johns Hopkins University Department of Pathology 410-614-0915 ------------------------------ Message: 5 Date: Mon, 15 Aug 2005 10:14:35 -0400 From: "Kopczynski, Charlotte" Subject: [Histonet] Breast Implant Storage To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <5452D66669CFC540B0452115ED26AF1F0191AEFC@bcexch02.bcad.baycare.org> I'm interested in how long you are keeping breast implants that have been removed. I have seen the recommendations on the CAP site but they do not address the length of time these specimens should be kept. I am trying to get my Risk management Department to make a decision. Thanks, Charlotte Kopczynski,HTL(ASCP) Baycare Pathology Manager Phone: 727-461-8246 Fax: 727-462-7597 Confidential: This electronic message and all contents contain information from BayCare Health System which may be privileged, confidential or otherwise protected from disclosure. The information is intended to be for the addressee only. If you are not the addressee, any disclosure, copy, distribution or use of the contents of this message is prohibited. If you have received this electronic message in error, please notify the sender and destroy the original message and all copies. ------------------------------ Message: 6 Date: Mon, 15 Aug 2005 09:56:21 -0500 From: "Denise Van Eaton" Subject: [Histonet] Shandon Citadel 2000 for sale To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed We have 3, due to some reinstrumentation. Ages are 1983, 1993,and 1996. All three are the 2000 model, no vacuum.Two of the three are in good working order, the third is missing a paraffin bath (cracked). Let us know, we'd sure like to find them all a new home! thanks,denise _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ ------------------------------ Message: 7 Date: Mon, 15 Aug 2005 08:00:24 -0700 From: "Laurie Colbert" Subject: RE: [Histonet] Breast Implant Storage To: "Kopczynski, Charlotte" , "Histonet (E-mail)" Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628005653744@EXCHANGE1.huntingtonhospital.com> Content-Type: text/plain; charset="iso-8859-1" Six months - the same as hardware. We used to keep them indefinitely, but you can imagine what a mess that was. Laurie Colbert -----Original Message----- From: Kopczynski, Charlotte [mailto:Charlotte.Kopczynski@baycare.org] Sent: Monday, August 15, 2005 7:15 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Breast Implant Storage I'm interested in how long you are keeping breast implants that have been removed. I have seen the recommendations on the CAP site but they do not address the length of time these specimens should be kept. I am trying to get my Risk management Department to make a decision. Thanks, Charlotte Kopczynski,HTL(ASCP) Baycare Pathology Manager Phone: 727-461-8246 Fax: 727-462-7597 Confidential: This electronic message and all contents contain information from BayCare Health System which may be privileged, confidential or otherwise protected from disclosure. The information is intended to be for the addressee only. If you are not the addressee, any disclosure, copy, distribution or use of the contents of this message is prohibited. If you have received this electronic message in error, please notify the sender and destroy the original message and all copies. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Mon, 15 Aug 2005 11:38:55 EDT From: RCHIOVETTI@aol.com Subject: Re: [Histonet] Nikon relay lens To: nyilmaz@mersin.edu.tr, histonet@lists.utsouthwestern.edu Message-ID: <1c3.2eb8ab83.3032110f@aol.com> Content-Type: text/plain; charset="US-ASCII" In a message dated 8/14/2005 11:35:35 PM US Mountain Standard Time, nyilmaz@mersin.edu.tr writes: > We're using an Olympus BX 50 light microscope attached with Nikon Coolpix > 5000 digital camera. I'm wondering what the mangnification value of MDC relay > lens between the camera and the microscope is. Dr. Yilmaz, I recently set up a system on a Leica microscope, using a Nikon Coolpix 8800. We did the following: 1. We obtained a stage micrometer, which is a precise ruling that is etched on a microscope slide. The one we used was 1 mm overall length, with smallest rulings of 0.010 mm (10 micrometers). 2. We did not use the MDC relay lens, but we purchased an adapter from another company. 3. On our system, we had to use the optical zoom of the camera to obtain a full field of view (eliminate the vignetting, or "clipping" of the image at the corners). 4. Since the zoom on the camera is infinitely adjustable, we had to decide on a reproducible zoom setting that always gave the same magnification. For the Coolpix 8800 and our adapter, we opted to zoom to the maximum setting. It's 10x on the 8800; I'm not sure what it is on the 5000. 5. To make measurements we set the camera to this maximum (10x) zoom, and took pictures of the stage micrometer with each objective. From there, it's relatively easy to measure the spacing of the 10 micrometer lines and to calculate a total magnification. This was the only way we could come up with to make measurements, since the camera's zoom is so variable. It's a small inconvenience, but the camera works great with this set-up. You could probably use a similar procedure, provided you are using the camera's optical zoom to fill the field of view with the MDC relay lens. The trick is to use a zoom setting that is reproducible. Hope this is of some help. Best regards, Robert (Bob) Chiovetti, Ph.D. The Microscope Works Arizona's Microscopy Resource 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA ------------------------------ Message: 9 Date: Mon, 15 Aug 2005 10:50:19 -0500 From: histology@gradymem.org Subject: [Histonet] handling fee specimens/tests sent to ref. lab To: Histonet Message-ID: <2b50526ed7.26ed72b505@onenet.net> Content-Type: text/plain; charset=us-ascii Does anyone charge the patient a handling fee for specimens sent to a reference lab? We send our cytologies out and used to charge a handling fee. Any comments? Angie ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 21, Issue 19 **************************************** The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. From TJJ <@t> Stowers-Institute.org Tue Aug 16 11:28:59 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Re: Immunofluorescence problem - Non-specific binding appears in control slide Message-ID: Nick, Quite the dilemma! Thanks for your detailed information on what you're seeing and what you've tried. I find this question interesting because at the moment I am sifting through a similar dilemma with regards to immunofluorescence and chromogenic immunostaining on mouse intestine. We have done immunostaining using an anti-BrdU mouse monoclonal antibody in mouse tissues. Routine technique includes 3% H2O2 for 10 minutes to block endogenous peroxidase, PowerBlock (biogenex) as a blocking reagent prior to primary antibody incubation, and avidin/biotin block (Vector) when using any detection involving biotin. Using an anti-mouse secondary antibody gives us strong staining with some intensely stained cells in the lamina propria, even in the negative sample (non-immune serum). We see this on both chromogenic (DAB) and fluorescent immuno procedures. Thinking it was a reaction with the anti-mouse secondary antibody (plasma cells or mast cells), we plugged this BrdU antibody in to the Dako ARK kit (biotinylates the primary + adds blockers and uses streptavidin/HRP) and also the Innogenex mouse on mouse kit. Believe it or not, we still got strong staining of these same cells on the negative control and also on the BrdU antibody sample. At the same time, we also tried using an anti-IgG2a anti-mouse/HRP labeled secondary antibody, and in the negative control there was no staining, while in the BrdU antibody sample there was a notable reduction in the number of positive cells stained in the lamina propria. Since we're using DAB chromogen and not fluorescence for this particular study, I don't think the SIF cell explanation fits here. This phenomena is not completely due to anti-mouse secondaries because we still see the reaction with the biotin/streptavidin detection. It isn't endogenous peroxidase because it is eradicated using an isotype-specific secondary on negative control, however we do see reactivity when the BrdU antibody is used. Consequently we've adopted using the anti-IgG2a secondary antibody as a "cure" to this problem, but I'm still curious about what's really going on! Perhaps it's a sensitivity issue with regards to the detection scheme (negative controls), but the isotype specific secondary antibody gives us staining in the BrdU antibody samples (specific staining?) I am in search of answers to this dilemma, so if anybody has any suggestions, I'm open to hearing them. I suspect you're seeing the same cells staining that we are, but additionally you have some background issues with connective tissue and muscle. Aldehyde-fixed tissue tends to give high auto-fluorescence, especially in muscle and red blood cells. You might try using the combination of acetone/alcohol (3 parts/1part) as a fixative for your cryosectioned tissues to improve morphology and take the aldehyde fixation out of the picture. Here is the technique from Gayle Callis: An acetone/alcohol fixation (AA) Air dry frozen sections overnight at RT, they MUST BE VERY DRY. Fix next day (of staining) in 75% acetone/25% absolute ethanol or 75 ml acetone mixed with 25 ml 100% ethanol (do NOT use methanol or any other alcohol) . Fix for 5 minutes, then go directly to buffer rinse. DO NOT air dry sections again, and proceed to staining. Otherwise you might try any of the published techniques for abolishing auto-fluorescence. Many anecdotal reviews are mixed on how effective these really are. And, for the record, immunostaining with primary antibodies made in goat is one of my LEAST favorite things to do. I would recommend concentrating on using the rabbit polyclonal antibody for future development. Would a silver reaction (special stain) work for your purposes to identify these cells instead? Best wishes, Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From shu-cheng.chen <@t> spcorp.com Tue Aug 16 12:05:51 2005 From: shu-cheng.chen <@t> spcorp.com (Chen, Shu-Cheng) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] research job posting at NJ Message-ID: Hi, We have the following position available at our organization. Anyone interested please respond to this mail. Or, apply on line at http://www.schering-plough.com/schering_plough/careers/jobframe.jsp . Thanks. Shu-Cheng Chen SPRI Kenilworth, NJ Position Title: Senior Scientist II Business: Division Research and Development Location: NJ Kenilworth Job Description: We are seeking a conscientious and experienced scientist with a BS or MS and more than 5 years experience to join the Inflammation Department at the Schering Plough Research Institute. Expertise in tissue processing, embedding, paraffin and frozen tissue section, histological staining, immunohistochemistry and in situ hybridization is required. Experience with both rodent and human tissues is desired. Additional experience and knowledge in necropsy, in vivo models with relevance to human inflammatory diseases and handling of rodents is preferred. ********************************************************************* 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 gcallis <@t> montana.edu Tue Aug 16 12:15:47 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] removing debris Re: Grinding & polishing of resin sections. In-Reply-To: <194.4500e299.30336864@aol.com> References: <194.4500e299.30336864@aol.com> Message-ID: <6.0.0.22.1.20050816105424.01b6a858@gemini.msu.montana.edu> Sulekha, am assuming you are doing thick slab type sections with grinding/polished and possible surface type staining? To remove debris from ground sections, immerse sections into water with a drop of detergent in a beaker, ultrasonicate section - you will actually see the debris fly off the surface. Be sure to rinse with distilled water or simply hold the section under a hard stream of tap water to rinse off residual detergent. We did this with all our slab MMA bone sections but you MUST use water based alumina polishing compound as a final polish instead of diamond paste if the paste contains oil lubricants, not good!! Alumina is also MUCH cheaper. Oil lubricants are difficult to remove unless you use some organic solvent and you may not want to etch the resin with a solvent just to clean off debris. We would grind with 640 grit paper then go directly to 1 um size alumina, you can achieve a fine polished surface and remove any unsightly scratches which pick up stain. There is a 0.1 um and 0.3 um size alumina, but we rarely used these. We had a Buehler grinder polisher, started with 320 grit paper, went to 640 then 1 um alumina polish compound. Depending on what grit size your diamond cut off blade has on it edge, that determines the next grit you would go to to start grinding. So if the blade had 240 grit diamond embedded in the edge, then you can proceed to the next grit on grinding paper to begin final grinding. If you use too low a grit number you can actually grind too much specimen away, ruining a lesion or area you want to see. I would want to remove this fine debris pushed into little spaces (they are there!) before removing plastic too, it could continue to float around in staining solutions. It is wise to wash between each grit, so you are not retaining big grit before the next paper grit size. At 10:03 AM 8/16/2005, you wrote: >In a message dated 8/16/2005 2:00:20 AM US Mountain Standard Time, >sulekhababy@yahoo.co.uk writes: > > > After grinding &polishing using grinding papers and diamond paste of > > different sizes we find the sections contain lots of debris.What can I > do to clean > > these resin sections before it is stained? > > > >Sulekha, > >Can you completely remove the plastic from the sections before staining? >This would certainly remove the bulk of the contaminants, since they're >getting >embedded in the sections during the polishing process. > >It would be similar to the deparaffinization of paraffin sections. > >You could probably use 2-methoxy ethyl acetate to remove the methacrylate >plastic. Then you would simply have the specimens remaining on the slide, >with >no plastic Wash and rehydrate thoroughly after the "deacrylation," and >proceed >with staining. > >See the following link: >< >http://www.ecmjournal.org/journal/supplements/vol007supp01/pdf/vol007supp01a82.pdf>. >Robert (Bob) Chiovetti, Ph.D. >The Microscope Works >Arizona's Microscopy Resource >132 North Elster Drive >Tucson, AZ 85710-3212 USA >Tel./Fax 520-546-4986 >Member, Arizona Small Business Association - ASBA Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Tue Aug 16 12:51:43 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:27 2005 Subject: [Histonet] Re: Immunofluorescence problem - Non-specific binding appears in control slide In-Reply-To: <200508161635.j7GGZhKM019296@mail1.msu.montana.edu> References: <200508161635.j7GGZhKM019296@mail1.msu.montana.edu> Message-ID: <6.0.0.22.1.20050816111706.01b47010@gemini.msu.montana.edu> Teri and Nick, Just curious about which block you use with Strepavidin-Biotin methods? We use the Strepavidin/biotin block from Vector instead of their avidin/biotin blocker when working with Strepavidin-Alexa fluorophores and Strepavidin/biotin for IHC. I asked the Vector technical service contact and this was his reply (some time ago) Thanks for the email. Streptavidin does have some subtle binding differences than avidin. Streptavidin has been shown to bind to fibronectin receptors that are expressed on cell types. These integrins (extracellular matrix receptors) mediate cell adhesion, migration etc and streptavidin binds to these too. So the bottomline this that streptavidin based detection systems may give background/non-specific staining due to this inetgrin binding that is not blocked by a standard avidin/biotin blocking step. Hence the idea behind the streptavidin/biotin kit. See the following reference: Alon, R. et al (1991) Cell-adhesive properties of streptavidin are mediated by the exposure of an RGD-like RYD site. Eur. J. Cell Biol. 58:271-279. The nomenclature in this article is dated. The GpIIbIIIa is now known as the alpha 5 beta 1 dimer that specifically binds fibronectin via the RGD amino acid sequence. There are antibodies available against the alpha 5 subunit itself. Migratory cells, such as embryonic cells, some tumor cells etc would be high expressors of this integrin. Obviously you could either block with the streptavidin kit or use an avidin based detection system. *** We have found the Strepavidin-biotin blocking kit works well, things remain clean with both IFA and IHC. Teri made a good suggestion to use a secondary against the specific isotype. With IHC, have you thought of using the glucose oxidase endogenous peroxidase blocking method, very thorough and kills both peroxidases and pseudoperoxidases, works with frozen sections and with paraffin sections. I will be happy to send this to you, we found it took out a cells staining nonspecfically positive in our negative control (not good!) and something you don't want when the target cell population is sparse to very fee then you want NO spurious nonspecific staining of any cells. GLUOX as we called it cleaned up a multitude of staining sins. There will always be some autofluorescence in the tissue, even after acetone alcohol or acetone fixation of frozen sections. One thing to consider is using a fluorophore that contrasts the autofluorescence, in red rather than FITC colors - we use Alexa 555 to do this, then autofluorescence becomes a counterfluorescence - making interesting photos!!! The strength of the signal of your fluorophore will help, something that is extremely bright (Alexa 488 versus FITC) and overcomes some of the autofluorence also helps. We also are picky where we purchase our fluorophore labelled antibodies, by F(ab')2 frag of IgG to avoid fc receptor binding in tissue, and careful adsorption to the mouse or species being stained. We have never tried methods to abolish autofluroescence, in general, we live with it or deal with it in a different way. At 10:28 AM 8/16/2005, you wrote: >Nick, > >Quite the dilemma! Thanks for your detailed information on what you're >seeing and what you've tried. I find this question interesting because >at the moment I am sifting through a similar dilemma with regards to >immunofluorescence and chromogenic immunostaining on mouse intestine. > >We have done immunostaining using an anti-BrdU mouse monoclonal antibody >in mouse tissues. Routine technique includes 3% H2O2 for 10 minutes to >block endogenous peroxidase, PowerBlock (biogenex) as a blocking reagent >prior to primary antibody incubation, and avidin/biotin block (Vector) >when using any detection involving biotin. > >Using an anti-mouse secondary antibody gives us strong staining with >some intensely stained cells in the lamina propria, even in the negative >sample (non-immune serum). We see this on both chromogenic (DAB) and >fluorescent immuno procedures. Thinking it was a reaction with the >anti-mouse secondary antibody (plasma cells or mast cells), we plugged >this BrdU antibody in to the Dako ARK kit (biotinylates the primary + >adds blockers and uses streptavidin/HRP) and also the Innogenex mouse on >mouse kit. Believe it or not, we still got strong staining of these >same cells on the negative control and also on the BrdU antibody sample. >At the same time, we also tried using an anti-IgG2a anti-mouse/HRP >labeled secondary antibody, and in the negative control there was no >staining, while in the BrdU antibody sample there was a notable >reduction in the number of positive cells stained in the lamina propria. >Since we're using DAB chromogen and not fluorescence for this particular >study, I don't think the SIF cell explanation fits here. This phenomena >is not completely due to anti-mouse secondaries because we still see the >reaction with the biotin/streptavidin detection. It isn't endogenous >peroxidase because it is eradicated using an isotype-specific secondary >on negative control, however we do see reactivity when the BrdU antibody >is used. > >Consequently we've adopted using the anti-IgG2a secondary antibody as a >"cure" to this problem, but I'm still curious about what's really going >on! Perhaps it's a sensitivity issue with regards to the detection >scheme (negative controls), but the isotype specific secondary antibody >gives us staining in the BrdU antibody samples (specific staining?) I >am in search of answers to this dilemma, so if anybody has any >suggestions, I'm open to hearing them. > >I suspect you're seeing the same cells staining that we are, but >additionally you have some background issues with connective tissue and >muscle. Aldehyde-fixed tissue tends to give high auto-fluorescence, >especially in muscle and red blood cells. You might try using the >combination of acetone/alcohol (3 parts/1part) as a fixative for your >cryosectioned tissues to improve morphology and take the aldehyde >fixation out of the picture. Here is the technique from Gayle Callis: >An acetone/alcohol fixation (AA) Air dry frozen sections overnight at >RT, they MUST >BE VERY DRY. Fix next day (of staining) in 75% acetone/25% absolute >ethanol or 75 ml acetone mixed with 25 ml 100% ethanol (do NOT use >methanol >or any other alcohol) . Fix for 5 minutes, then go directly to buffer >rinse. DO NOT air dry sections again, and proceed to staining. > >Otherwise you might try any of the published techniques for abolishing >auto-fluorescence. Many anecdotal reviews are mixed on how effective >these really are. > >And, for the record, immunostaining with primary antibodies made in goat >is one of my LEAST favorite things to do. I would recommend >concentrating on using the rabbit polyclonal antibody for future >development. > >Would a silver reaction (special stain) work for your purposes to >identify these cells instead? > >Best wishes, > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, MO 64133 > >_______________________________________________ >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 Barry.R.Rittman <@t> uth.tmc.edu Tue Aug 16 13:19:02 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] removing debris Re: Grinding & polishing of resinsections. Message-ID: It is a long time since I polished ground sections (usually without the benefits of a Buehler polishing machine), and I totally agree with Gayle's comments. Another point worth mentioning perhaps is that several of the diamond pastes are colored to prevent mixing up grades I assume. This dye plus the oil base tends to stain some substances in the slices whether they are unembedded or in plastic. The same is also true for polishing rocks slices. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Tuesday, August 16, 2005 12:16 PM To: RCHIOVETTI@aol.com; Histonet@lists.utsouthwestern.edu Subject: [Histonet] removing debris Re: Grinding & polishing of resinsections. Sulekha, am assuming you are doing thick slab type sections with grinding/polished and possible surface type staining? To remove debris from ground sections, immerse sections into water with a drop of detergent in a beaker, ultrasonicate section - you will actually see the debris fly off the surface. Be sure to rinse with distilled water or simply hold the section under a hard stream of tap water to rinse off residual detergent. We did this with all our slab MMA bone sections but you MUST use water based alumina polishing compound as a final polish instead of diamond paste if the paste contains oil lubricants, not good!! Alumina is also MUCH cheaper. Oil lubricants are difficult to remove unless you use some organic solvent and you may not want to etch the resin with a solvent just to clean off debris. We would grind with 640 grit paper then go directly to 1 um size alumina, you can achieve a fine polished surface and remove any unsightly scratches which pick up stain. There is a 0.1 um and 0.3 um size alumina, but we rarely used these. We had a Buehler grinder polisher, started with 320 grit paper, went to 640 then 1 um alumina polish compound. Depending on what grit size your diamond cut off blade has on it edge, that determines the next grit you would go to to start grinding. So if the blade had 240 grit diamond embedded in the edge, then you can proceed to the next grit on grinding paper to begin final grinding. If you use too low a grit number you can actually grind too much specimen away, ruining a lesion or area you want to see. I would want to remove this fine debris pushed into little spaces (they are there!) before removing plastic too, it could continue to float around in staining solutions. It is wise to wash between each grit, so you are not retaining big grit before the next paper grit size. At 10:03 AM 8/16/2005, you wrote: >In a message dated 8/16/2005 2:00:20 AM US Mountain Standard Time, >sulekhababy@yahoo.co.uk writes: > > > After grinding &polishing using grinding papers and diamond paste of > > different sizes we find the sections contain lots of debris.What can I > do to clean > > these resin sections before it is stained? > > > >Sulekha, > >Can you completely remove the plastic from the sections before staining? >This would certainly remove the bulk of the contaminants, since they're >getting >embedded in the sections during the polishing process. > >It would be similar to the deparaffinization of paraffin sections. > >You could probably use 2-methoxy ethyl acetate to remove the methacrylate >plastic. Then you would simply have the specimens remaining on the slide, >with >no plastic Wash and rehydrate thoroughly after the "deacrylation," and >proceed >with staining. > >See the following link: >< >http://www.ecmjournal.org/journal/supplements/vol007supp01/pdf/vol007su pp01a82.pdf>. >Robert (Bob) Chiovetti, Ph.D. >The Microscope Works >Arizona's Microscopy Resource >132 North Elster Drive >Tucson, AZ 85710-3212 USA >Tel./Fax 520-546-4986 >Member, Arizona Small Business Association - ASBA 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 AMH-HISTOLAB <@t> amh.org Tue Aug 16 13:30:55 2005 From: AMH-HISTOLAB <@t> amh.org (AMH-HistoLab) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] histology supervisor position in PA Message-ID: ABINGTON MEMORIAL HOSPITAL IS LOOKING FOR A FULL TIME HISTOLOGY SUPERVISOR. PLEASE CONTACT: AMH.ORG TO APPLY ON LINE OR CALL 215-481-2650. ****************** CONFIDENTIALITY NOTICE ********************** This e-mail contains LEGALLY PRIVILEGED AND CONFIDENTIAL INFORMATION intended only for the use of the recipient named above. If you are not the intended recipient, you are hereby notified that any dissemination or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please notify the transmitting hospital by telephone or e-mail and delete the original e-mail received in error. THIS INFORMATION HAS BEEN DISCLOSED TO YOU FROM RECORDS WHOSE CONFIDENTIALITY IS PROTECTED BY STATE AND FEDERAL LAW. ANY FURTHER DISCLOSURE, COPYING, DISTRIBUTION OR ACTION TAKEN IN RELIANCE ON THE CONTENTS OF THESE DOCUMENTS WITHOUT THE PRIOR WRITTEN CONSENT OF THE PERSON TO WHOM IT PERTAINS IS PROHIBITED. YOU ARE REQUIRED TO DESTROY THE INFORMATION AFTER THE STATED NEED HAS BEEN FULFILLED. From DOOLEEO <@t> shands.ufl.edu Tue Aug 16 14:14:29 2005 From: DOOLEEO <@t> shands.ufl.edu (Elaine Dooley) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Beta F1 Message-ID: Dear Histonetters, One of our hemepath pathologists wants us to get an antibody called Beta F1. Is there a vendor that has this antibody? If so I need one that works on formalin fixed paraffin embedded tissue. Thanks in advance Elaine Dooley 352-265-0111 ext 72117 Shands Teaching Hospital Gainesville FL From THOMASEBROOKS <@t> aol.com Tue Aug 16 15:27:36 2005 From: THOMASEBROOKS <@t> aol.com (THOMASEBROOKS@aol.com) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Beta F1 Message-ID: <1d9.42a152fd.3033a638@aol.com> Elaine, You are probably referring to Human TCR beta F1 MAb (clone 8A3), Product code TCR1151, from Pierce. Pierce Biotechnology, Inc. Customer Service Department P.O. Box 117 Rockford, IL. 61105 (800) 874-3723 Thomas Brooks From rnewlin <@t> burnham.org Tue Aug 16 16:31:38 2005 From: rnewlin <@t> burnham.org (Robin Newlin) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] aldehyde fuchsin Message-ID: <005f01c5a2a9$e2f50b90$386810ac@RNEWLIN> Hi All Does anyone know if aldehyde fuchsin is available commercially, or do I have to brew my own. Thanks Robbin Newlin Manager, Cell Imaging/Histology Core Facility The Burnham Institute 10901 N. Torrey Pines Road LaJolla, Ca. 92037 858-646-3100 ext.3552 From Robert.Lott <@t> bhsala.com Tue Aug 16 17:51:56 2005 From: Robert.Lott <@t> bhsala.com (Lott, Robert) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Re: Endogenous peroxidase Message-ID: Jim, ...perhaps it is "sodium azide" rather than sodium nitride???? Hydrogen peroxide and sodium azide together in solution is very effective in eliminating "endogenous peroxidase" Robert Robert L. Lott, HTL(ASCP) Baptist Health System 800 Montclair Road Birmingham, AL 35213 205-592-5388 phone 205-592-5646 fax robert.lott@bhsala.com From: histonet-bounces@lists.utsouthwestern.edu on behalf of Mr James Hugh Reilly Sent: Tuesday, August 16, 2005 12:28 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Endogenous Peroxidase I have been asked to try a new Immumohistochemistry (IHC) technique which uses 1% hydrogen peroxide and 2% sodium nitride in phosphate buffer solution. Does anyone know what sodium nitride Is? and where can it be purchased? Jim Reilly University Dept. Medicine Level 3 QEB Royal Infirmary 10 Alexandra Parade Glasgow G31 2ER Message: 10 Date: Tue, 16 Aug 2005 09:41:57 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] Endogenous Peroxidase To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175AE@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" Sodium nitride is Na3N, but I don't think you will be able to purchase it anywhere. It is a compound that was long believed to be chemically impossible, but which only recently was experimentally synthesized for the first time by deposition of atomic sodium and nitrogen beams onto a supercooled sapphire in a vacuum. I doubt that the compound is commercially available. Perhaps it is sodium nitrite that is called for in your protocol? Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Mr > James Hugh Reilly > Sent: Tuesday, August 16, 2005 12:28 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Endogenous Peroxidase > > I have been asked to try a new Immumohistochemistry (IHC) technique which > uses 1% hydrogen peroxide and 2% sodium nitride in phosphate buffer > solution. Does anyone know what sodium nitride Is? and where can it be > purchased? > > Jim Reilly > > University Dept. Medicine > Level 3 QEB > Royal Infirmary > 10 Alexandra Parade > Glasgow > G31 2ER > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 11 Date: Tue, 16 Aug 2005 09:51:48 -0400 From: "John A. Kiernan" Subject: Re: [Histonet] Endogenous Peroxidase To: Mr James Hugh Reilly Cc: histonet@lists.utsouthwestern.edu Message-ID: <4301EF74.DCDF1755@uwo.ca> Content-Type: text/plain; charset=us-ascii I think there must be a mistake in the name. Sodium nitride (Na3N) is a grey solid formed by heating metallic sodium in nitrogen. It is instantly decomposed by water, so a solution in phosphate buffer could not be made. -- ------------------------------- 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 _______________________________ Mr James Hugh Reilly wrote: > > I have been asked to try a new Immumohistochemistry (IHC) technique which uses 1% hydrogen peroxide and 2% sodium nitride in phosphate buffer solution. Does anyone know what sodium nitride Is? and where can it be purchased? > > Jim Reilly > > University Dept. Medicine > Level 3 QEB > Royal Infirmary > 10 Alexandra Parade > Glasgow > G31 2ER > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Tue, 16 Aug 2005 09:52:14 -0400 From: "Smith, Allen" Subject: RE: [Histonet] Endogenous Peroxidase To: "Mr James Hugh Reilly" Cc: histonet@lists.utsouthwestern.edu Message-ID: <5D2189E74151CC42BEC02906BA8996322B90DD@exchsrv01.barrynet.barry.edu> Content-Type: text/plain; charset="us-ascii" Sodium nitride, Na3N, is a rather unstable compound first prepared in 2002. The synthesis is notoriously difficult. I do not think it is commercially available. I think that the "nitride" in your recipe is a typographical error in "nitrite". Sodium nitrite is available from both Aldrich and Fisher. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mr James Hugh Reilly Sent: Tuesday, August 16, 2005 3:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Endogenous Peroxidase I have been asked to try a new Immumohistochemistry (IHC) technique which uses 1% hydrogen peroxide and 2% sodium nitride in phosphate buffer solution. Does anyone know what sodium nitride Is? and where can it be purchased? Jim Reilly University Dept. Medicine Level 3 QEB Royal Infirmary 10 Alexandra Parade Glasgow G31 2ER Confidentiality Notice: The information contained in this email message is privileged and confidential information and intended only for the use of the individual or entity named in the address. If you are not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this information is strictly prohibited. If you received this information in error, please notify the sender and delete this information from your computer and retain no copies of any of this information. From ckeith71 <@t> hotmail.com Wed Aug 17 02:46:58 2005 From: ckeith71 <@t> hotmail.com (cindy keith) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Help with Processing run Message-ID: I started a new job in a lab that has always only had a day shift. My new hours here are 3 am -11am replacing a person who worked 530am-2pm. This change was not well received by my fellow coworkers/supervisor. My question is if the processing run ends at 3am and if I were to be out sick would you think it would be ok to leave bx/lg tissue sitting in the parrafin at 62-64 degrees for 2 1/2 hours-to potentially 3 hours waiting for the next person to come in or would you think it would be better for the tissue to be removed by a fellow lab person and hardened on counter then remelted to embed upon someone else arriving? Immunos are performed on this tissue also. Your responses would be greatly appreciated. From c.m.vanderloos <@t> amc.uva.nl Wed Aug 17 03:10:34 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] RE: Endogenous Peroxidase Message-ID: <32319d324852.32485232319d@amc.uva.nl> Hello James and others, Despite of all the (useless) chemical stories you've received so far I think that there's mix-up of Na3N and NaN3 here. "Sodium nitride" should be "sodium azide" = NaN3. That would fit in very well with your subject line: "endogenous peroxidase". The mixture of PBS (or TBS) with Na-azide and peroxide is a well known procedure to quench endogenous peroxidase activity (Li et al., JHC 35:1457-1460, 1987). The only thing that doesn't fit here is the fact it isn't new... Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Tue, 16 Aug 2005 07:28:36 +0000 From: "Mr James Hugh Reilly" Subject: [Histonet] Endogenous Peroxidase To: histonet@lists.utsouthwestern.edu I have been asked to try a new Immumohistochemistry (IHC) technique which uses 1% hydrogen peroxide and 2% sodium nitride in phosphate buffer solution. Does anyone know what sodium nitride Is? and where can it be purchased? Jim Reilly University Dept. Medicine Level 3 QEB Royal Infirmary 10 Alexandra Parade Glasgow G31 2ER From Megan.Clarke <@t> hnehealth.nsw.gov.au Wed Aug 17 03:23:26 2005 From: Megan.Clarke <@t> hnehealth.nsw.gov.au (Megan Clarke) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] mitochondrial antibody enquiry and DAB waste disposal Message-ID: Hi. I have 2 queries. Firstly, could anyone please assist me in finding a supplier for anti- mitochondrial antibody for use in FFPET (formalin fixed paraffin embedded tisue) and secondly, with regard to DAB waste disposal, is the potassium permanganate/sulphuric acid method still the one of choice for neutralizing DAB or is there some new magic available? If their are any BONDMAX users out there, I would appreciate it if you could share your DAB waste disposal method with me. Thanking you Z Haffajee/Megan Clarke HAPS IHC unit Newcastle From Kemlo.Rogerson <@t> elht.nhs.uk Wed Aug 17 03:35:25 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Help with Processing run Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4EC@elht-exch1.xelht.nhs.uk> Personally, no, but the problem sits with your Operational Procedures. Both they and ICC require standardisation and having differing wax impregnation time's flies in the face of good laboratory practice. Hardening then re-melting is an answer but that will delay blocking out until the wax had melted again. Maybe the answer is to never be sick or have a backup processing cycle that finishes 3 hours later; I concede that will not accommodate your first day of sickness but then there would not be anyone there to take the biopsies out of wax to harden anyway; would there? You need a processing machine connected to the internet or dial up that can be altered remotely; Biochemistry has them! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of cindy keith Sent: 17 August 2005 08:47 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Help with Processing run I started a new job in a lab that has always only had a day shift. My new hours here are 3 am -11am replacing a person who worked 530am-2pm. This change was not well received by my fellow coworkers/supervisor. My question is if the processing run ends at 3am and if I were to be out sick would you think it would be ok to leave bx/lg tissue sitting in the parrafin at 62-64 degrees for 2 1/2 hours-to potentially 3 hours waiting for the next person to come in or would you think it would be better for the tissue to be removed by a fellow lab person and hardened on counter then remelted to embed upon someone else arriving? Immunos are performed on this tissue also. Your responses would be greatly appreciated. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kappeler <@t> patho.unibe.ch Wed Aug 17 04:57:03 2005 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] mitochondrial antibody enquiry and DAB waste disposal References: Message-ID: <008601c5a312$16e1e910$27955c82@patho.unibe.ch> Hi Megan mo-a-Mitochondrial Antigen, clone 113-1, BioGenex CatNo MU213-UC, works reasonably well on FFPE tissue. We had best results after HIER (citrate) in a pressure cooker and with the mAb diluted at 1:50 (which is a relatively useless information, however BioGenex is also one of those companies that do not specify Ig content in ug/ml for their antibodies, therefore I have to give you the information '1:50' rather than x ug Ig/ml). Hope this helps Andi Kappeler Institute of Pathology, University of Bern, Switzerland ----- Original Message ----- From: "Megan Clarke" To: Cc: Sent: Wednesday, August 17, 2005 10:23 AM Subject: [Histonet] mitochondrial antibody enquiry and DAB waste disposal > Hi. I have 2 queries. > > Firstly, could anyone please assist me in finding a supplier for anti- > mitochondrial antibody for use in FFPET (formalin fixed paraffin embedded > tisue) and > > secondly, with regard to DAB waste disposal, is the potassium > permanganate/sulphuric acid method still the one of choice for > neutralizing DAB or is there some new magic available? If their are any > BONDMAX users out there, I would appreciate it if you could share your > DAB waste disposal method with me. > > Thanking you > > Z Haffajee/Megan Clarke > HAPS > IHC unit > Newcastle > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From DDittus787 <@t> aol.com Wed Aug 17 08:16:33 2005 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] cleaved notch antibody,and HIER Message-ID: <20a.73cbbab.303492b1@aol.com> Sorry to post this late, however I was reading this and thought some information may help, yes I agree that Zymeds anti goat anti rabbit and strepavidin are concentrates and dilution is important,however background is also from too long an incubation time or too high a titer,etc. So not only do you want to dilute the goat,rabbit but also the antibody and then running retrievals of some sort clouds troubleshooting. Change one part of the protocol at a time, prolonged fixation can cause background as well. I have used many Zymed products and have also found that their concentrations are high and you can diutle out pretty far as well as not have to incubate very long. Just some technical info Dana From DDDeltour <@t> mar.med.navy.mil Wed Aug 17 08:58:22 2005 From: DDDeltour <@t> mar.med.navy.mil (DDDeltour@mar.med.navy.mil) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Breast Processing Protocol Message-ID: <080566D001A3D9459FFC0A391A646C9106B1B19E@marxchg03.mar.med.navy.mil> I am looking for a program for processing fatty breast tissue. We currently let the breast sit in cassettes in Pen-Fix for an extra day before processing. We then process the first Fixative in Pen-Fix for one hour then in 10% Formalin for the second hour (I do not like this but that is they way it was done when I got here). The next steps are.. 95% one hour X3, 100% one hour X3, Xylene 1 hour X2. Paraffin for one hour X3. I am considering replacing one of the 95% with another fixative. What programs do you use for this? They don't like to hear Sh in Sh out:-). Thank you! DOUGLAS D. DELTOUR HISTOLOGY TECHNICIAN NMC PORTSMOUTH VA From bwhitaker <@t> brownpathology.com Wed Aug 17 09:05:45 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Help with Processing run In-Reply-To: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4EC@elht-exch1.xelht.nhs.uk> Message-ID: <000501c5a334$c3405fd0$3601a8c0@brownpathology.net> To play the devil's advocate: then what good is altering the processing schedule from home via internet ? Theoretically, changing the protocol to run 3 extra hours, regardless of what step(s) are lengthened, destandardizes the processing schedule. Of course, in reality, there will be times that the schedule is altered. As for how badly does it hurt the tissue to set for 3 additional hours in paraffin, I think that depends on the tissue. If they are large, fatty, mastectomy specimens, they will perhaps even benfit. If they are little GI bx's, then that's another ball of wax. If it is a biopsy run, then I do vote for getting someone to just remove the tissue and set it on the counter. Yes, it will delay it a little while it melts, but if you call in sick, they'll be running behind anyway, so what's a few more minutes. One quicker way to melt them (I think) is to submerse the cassettes in molten paraffin for a few minutes, and then put them in the embedding center (as opposed to just putting them in the embedding center to melt). Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rogerson Kemlo (ELHT) Pathology Sent: Wednesday, August 17, 2005 3:35 AM To: cindy keith; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Help with Processing run Personally, no, but the problem sits with your Operational Procedures. Both they and ICC require standardisation and having differing wax impregnation time's flies in the face of good laboratory practice. Hardening then re-melting is an answer but that will delay blocking out until the wax had melted again. Maybe the answer is to never be sick or have a backup processing cycle that finishes 3 hours later; I concede that will not accommodate your first day of sickness but then there would not be anyone there to take the biopsies out of wax to harden anyway; would there? You need a processing machine connected to the internet or dial up that can be altered remotely; Biochemistry has them! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of cindy keith Sent: 17 August 2005 08:47 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Help with Processing run I started a new job in a lab that has always only had a day shift. My new hours here are 3 am -11am replacing a person who worked 530am-2pm. This change was not well received by my fellow coworkers/supervisor. My question is if the processing run ends at 3am and if I were to be out sick would you think it would be ok to leave bx/lg tissue sitting in the parrafin at 62-64 degrees for 2 1/2 hours-to potentially 3 hours waiting for the next person to come in or would you think it would be better for the tissue to be removed by a fellow lab person and hardened on counter then remelted to embed upon someone else arriving? Immunos are performed on this tissue also. Your responses would be greatly appreciated. _______________________________________________ Histonet mailing list Histonet@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 MCKEELA <@t> UCMAIL.UC.EDU Wed Aug 17 09:27:00 2005 From: MCKEELA <@t> UCMAIL.UC.EDU (McKee, Lynn (mckeela)) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] unsubscribe Message-ID: <9871F0C990DF9B4681F1EB312A4D02F8443EC4@ucmail8.ad.uc.edu> From rbruggeman <@t> psu.edu Wed Aug 17 09:30:48 2005 From: rbruggeman <@t> psu.edu (Richard Bruggeman) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Rabbit Anti-sera Background Message-ID: We are having some problems with extensive background and nonspecific staining with a new rabbit polyclonal anti-sera. Here are some of the staining details: -FFPE human tissue -1mM EDTA pH 8.0 HIER for 20 minutes in steamer seems to provide best staining -10% H2O2 block for 10 minutes @ RT (standard for our lab) -block with 1% BSA in TBS for approx. 30 minutes @ RT (no rinse afterwards, just drain off blocking reagents) -primary antibody dilution worked out to 1:1000-2000 (diluted in Dako diluent) for 30 minutes @ RT via titration runs -Dako Envision+ Goat Anti-Rabbit/HRP Labeled polymer detection used for 30 minutes @ RT (standard for our lab) -Dako DAB+ for 10 minutes @ RT (standard for our lab) -counter stained w/ Dako Hematoxylin for 1-2 minutes (standard for our lab) -All rinses done in Dako TBS w/ 0.05% Dako Tween 20 (standard for our lab) We have tried doing peptide blocking experiments that have shown some of the staining can be blocked, however most of the staining is still present, suggesting that it is due to nonspecific binding. We know from Westerns that our blocking experiments include a VERY large excess of peptide, so all of the specific antibodies should be bound to peptide. Negative controls (without antibody and/or with peptide only) always show up DEAD negative. No avidin/biotin to worry about since this detections system is a labeled polymer. The detection system has been working very well for us in the past, including with other commercially available rabbit polyclonals, so I can't imagine this would be the cause of our problem. What suggestions might you have to reduce this intense background/nonspecific staining. 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 pex0220 <@t> yahoo.com.cn Wed Aug 17 09:48:55 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] IF in bone or cartilage paraffin sections Message-ID: <20050817144856.62764.qmail@web15509.mail.cnb.yahoo.com> Hello, all. I have done immunofluorescence in bone or cartilage paraffin sections for several months. I use normal immunofluorescence protocol for staining, but I have tried different methods for antigen retrieval, finally I think that 0.1% trypsin (15min or 20min) for antigen retrieval in my experiments is a little bit good. In addition, I have already used different antibodies for immunosatining. One of them is working well, positive expressions can be found in cartilage, osteoblasts and osteocytes, but others express in cartilage strongly, however they are very weak in bone( but they express stronger in primary osteoblasts).I do not know the reasons! Any suggestions will be useful for me! Thank you! Guofeng Guofeng --------------------------------- DO YOU YAHOO!? ÑÅ»¢Ãâ·ÑGÓÊÏ䣭ÖйúµÚÒ»¾øÎÞÀ¬»øÓʼþɧÈų¬´óÓÊÏä From jkiernan <@t> uwo.ca Wed Aug 17 09:50:41 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Re: Endogenous peroxidase References: Message-ID: <43034EC1.367DB7E3@uwo.ca> Robert Lott has surely hit the nail right on the head! Probably someone, somewhere, wrote Na3N (sodium nitride) when it should have been NaN3 (sodium azide). Taking notes and never discarding them is always virtuous, and virtue is often rewarding: occasionally to one's self, but usually to others. Robert's email to Histonet prompted a quick retrieval of a source: Li C-Y, Ziesmer SC, Lazcano-Villareal O (1986) Use of azide and hydrogen peroxide as an inhibitor for endogenous peroxidase in the immunoperoxidase method. J. Histochem. Cytochem. 35: 1457-1460. My notes say the solution contains 0.3% H2O2 and 0.1% NaN3 in phosphate buffered saline, applied for 10 minutes. Do not use the information in this this email without checking the original paper. Back issues of the J. Histochem. Cytochem. are available on the Web: http://www.jhc.org John Kiernan London, Canada ______________________________ From jkiernan <@t> uwo.ca Wed Aug 17 10:10:37 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] aldehyde fuchsin References: <005f01c5a2a9$e2f50b90$386810ac@RNEWLIN> Message-ID: <4303536D.D06870E2@uwo.ca> The solution of aldehyde fuchsine has a rather short shelf-life (8 weeks) so it's best to make up your own. It is also possible to recover the aldehyde fuchsine as a precipitate that can be collected by filtering, dried and stored for 10 or more years. Solutions made from aldehyde fuchsine powder (0.125% in 70% ethanol with 1% acetic acid) are more stable than the traditional directly made solution, being good for about 2 years. It is said that solutions of aldehyde fuchsine made from the powder will not stain pancreatic islet B cells without prior permanganate oxidation (see Mowry,RW 1978 Stain Technol. 53: 141-154. This paper also has useful tips for other uses of aldehyde fuchsine. -- ------------------------------- 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 _______________________________ Robin Newlin wrote: > > Hi All > Does anyone know if aldehyde fuchsin is available commercially, or do I have to brew my own. > Thanks > Robbin Newlin > Manager, Cell Imaging/Histology Core Facility > The Burnham Institute > 10901 N. Torrey Pines Road > LaJolla, Ca. 92037 > 858-646-3100 ext.3552 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tp2 <@t> medicine.wisc.edu Wed Aug 17 10:21:07 2005 From: tp2 <@t> medicine.wisc.edu (Thomas Pier) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] CD25 recipes Message-ID: Hello, I'm starting to work up CD25. I was just wondering if anyone had any good recipes or helpful tips, since I've heard that this can be tough one. Thanks. Tom Pier From Terry.Marshall <@t> rothgen.nhs.uk Wed Aug 17 10:50:25 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] aldehyde fuchsin Message-ID: John tells us: "It is also possible to recover the aldehyde fuchsine as a precipitate that can be collected by filtering, dried and stored for 10 or more years." OK John ......how? (This would solve my problem of needing it every 4 months only and suffering the groans of the techs at having to make it up again) BTW - I am sure that someone suggested a paraldehyde substitute at one time, but I omitted to keep the post. 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: John A. Kiernan [mailto:jkiernan@uwo.ca] Sent: 17 August 2005 16:11 To: Histonet Subject: Re: [Histonet] aldehyde fuchsin The solution of aldehyde fuchsine has a rather short shelf-life (8 weeks) so it's best to make up your own. It is also possible to recover the aldehyde fuchsine as a precipitate that can be collected by filtering, dried and stored for 10 or more years. Solutions made from aldehyde fuchsine powder (0.125% in 70% ethanol with 1% acetic acid) are more stable than the traditional directly made solution, being good for about 2 years. It is said that solutions of aldehyde fuchsine made from the powder will not stain pancreatic islet B cells without prior permanganate oxidation (see Mowry,RW 1978 Stain Technol. 53: 141-154. This paper also has useful tips for other uses of aldehyde fuchsine. -- ------------------------------- 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 _______________________________ Robin Newlin wrote: > > Hi All > Does anyone know if aldehyde fuchsin is available commercially, or do I have to brew my own. > Thanks > Robbin Newlin > Manager, Cell Imaging/Histology Core Facility > The Burnham Institute > 10901 N. Torrey Pines Road > LaJolla, Ca. 92037 > 858-646-3100 ext.3552 > _______________________________________________ > Histonet mailing list > Histonet@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 bhewlett <@t> cogeco.ca Wed Aug 17 10:54:45 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Rabbit Anti-sera Background References: Message-ID: <002901c5a344$32d5e770$6400a8c0@mainbox> Richard, I'll bet that this rabbit antibody is an Ig fraction of whole serum. As such, only up to about 10% of the available Ig's are directed against your antigen of interest. The remainder are directed against assorted other antigens, some of which may be human. 1) Try switching your protein block from 1% BSA to non-immune goat serum ( to match the secondary), 2) increase primary antibody dilution to say 1: 5000-10000 or more, and incubation time to overnight @ 4C ( dilutes out other naturally occuring antibodies which are usually at a lower concentration than your primary), 3) add 10-20% pooled human serum to diluted primary ( to adsorb anything against human proteins). If this doesn't work you may have to adsorb with tissue powders! Bryan ----- Original Message ----- From: "Richard Bruggeman" To: Sent: Wednesday, August 17, 2005 10:30 AM Subject: [Histonet] Rabbit Anti-sera Background > We are having some problems with extensive background and nonspecific > staining with a new rabbit polyclonal anti-sera. Here are some of the > staining details: > > -FFPE human tissue > -1mM EDTA pH 8.0 HIER for 20 minutes in steamer seems to provide best > staining > -10% H2O2 block for 10 minutes @ RT (standard for our lab) > -block with 1% BSA in TBS for approx. 30 minutes @ RT (no rinse > afterwards, just drain off blocking reagents) > -primary antibody dilution worked out to 1:1000-2000 (diluted in Dako > diluent) for 30 minutes @ RT via titration runs > > -Dako Envision+ Goat Anti-Rabbit/HRP Labeled polymer detection used for > 30 minutes @ RT (standard for our lab) > -Dako DAB+ for 10 minutes @ RT (standard for our lab) > -counter stained w/ Dako Hematoxylin for 1-2 minutes (standard for our > lab) > -All rinses done in Dako TBS w/ 0.05% Dako Tween 20 (standard for our > lab) > > > We have tried doing peptide blocking experiments that have shown some > of the staining can be blocked, however most of the staining is still > present, suggesting that it is due to nonspecific binding. We know from > Westerns that our blocking experiments include a VERY large excess of > peptide, so all of the specific antibodies should be bound to peptide. > Negative controls (without antibody and/or with peptide only) always > show up DEAD negative. No avidin/biotin to worry about since this > detections system is a labeled polymer. The detection system has been > working very well for us in the past, including with other commercially > available rabbit polyclonals, so I can't imagine this would be the cause > of our problem. What suggestions might you have to reduce this intense > background/nonspecific staining. > > 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 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gcallis <@t> montana.edu Wed Aug 17 11:05:01 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] making up aldehyde fuchsin with a substitute aldehyde In-Reply-To: <4303536D.D06870E2@uwo.ca> References: <005f01c5a2a9$e2f50b90$386810ac@RNEWLIN> <4303536D.D06870E2@uwo.ca> Message-ID: <6.0.0.22.1.20050817094847.01b49bb0@gemini.msu.montana.edu> One of the problems with making up aldehyde fuchsin is accessing paraldehyde, a controlled substance and requires a Drug Enforcement Administration number to purchase, payment of handling fee, and so on. If you are in a clinical lab and have the number in available pharmacy, no problem. For those who do not have a DEA number, then one can substitute acetaldehyde, an uncontrolled substance, very cheap, and but still breaks down after a year or so. Substitution is 2.5 mls acetaldehyde instear od 1.5 ml paraldehyde. This was published by Peggy Wenk, J Histotechnology, Letters to the Editor 19(4):353, 1996. This has worked well for her lab, and you can always contact her for further discussion. At 09:10 AM 8/17/2005, you wrote: >The solution of aldehyde fuchsine has a rather >short shelf-life (8 weeks) so it's best to make >up your own. It is also possible to recover >the aldehyde fuchsine as a precipitate that can >be collected by filtering, dried and stored >for 10 or more years. Solutions made from aldehyde >fuchsine powder (0.125% in 70% ethanol with >1% acetic acid) are more stable than the traditional >directly made solution, being good for about 2 years. > >It is said that solutions of aldehyde fuchsine >made from the powder will not stain pancreatic >islet B cells without prior permanganate oxidation >(see Mowry,RW 1978 Stain Technol. 53: 141-154. This >paper also has useful tips for other uses of >aldehyde fuchsine. >-- >------------------------------- >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 >_______________________________ >Robin Newlin wrote: > > > > Hi All > > Does anyone know if aldehyde fuchsin is available commercially, or do I > have to brew my own. > > Thanks > > Robbin Newlin > > Manager, Cell Imaging/Histology Core Facility > > The Burnham Institute > > 10901 N. Torrey Pines Road > > LaJolla, Ca. 92037 > > 858-646-3100 ext.3552 > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From pex0220 <@t> yahoo.com.cn Wed Aug 17 11:11:09 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Fix with 4% PFA Message-ID: <20050817161109.22589.qmail@web15504.mail.cnb.yahoo.com> Hello,all, Generally, organs are fixed for overnight or two days with 4% PFA, then they are embedded in the machine. If I would like to keep them longer, How long can organs( tissues) be fixed with 4% PFA in 4 degree room( but organs are not destroyed)? Thank you! Guofeng Guofeng --------------------------------- DO YOU YAHOO!? ÑÅ»¢ÓÊÏ䳬ǿÔöÖµ·þÎñ£­2G³¬´ó¿Õ¼ä¡¢pop3ÊÕÐÅ¡¢ÎÞÏÞÁ¿ÓʼþÌáÐÑ From David.Edmondson <@t> christie-tr.nwest.nhs.uk Wed Aug 17 11:16:12 2005 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Endogenous Peroxidase Message-ID: <3C83687E8F6AE04792E361ABE2D385B841800D@cht-mail2-2k.xchristie.nhs.uk> AND it is there as a bacteriocidal/fungicidal component, and does not do people of copper pipes much good either, as my memory has it. So we just went for using fresh buffer before there were any accidents. Dave Christie Manchester -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Mr James Hugh Reilly Sent: 16 August 2005 08:29 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Endogenous Peroxidase I have been asked to try a new Immumohistochemistry (IHC) technique which uses 1% hydrogen peroxide and 2% sodium nitride in phosphate buffer solution. Does anyone know what sodium nitride Is? and where can it be purchased? Jim Reilly University Dept. Medicine Level 3 QEB Royal Infirmary 10 Alexandra Parade Glasgow G31 2ER _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed Aug 17 11:36:17 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] aldehyde fuchsin References: Message-ID: <43036781.A575AFA@uwo.ca> This is the method in M. Gabe's "Histological Techniques" (1976); it's also in my textbook. A similar method was published by Rosa,CC 1953 Stain Technol. 28:299-302. For comparisons of aldehyde fuchsine made in different ways, see papers by GW Nettleton in J. Histochem. Cytochem. 1980-1982. Dissolve 1 g pararosaniline (CI 42500) in 200 ml water. Heat to boiling and leave to cool to room temperature. Add 2.0 ml concentrated hydrochloric acid. Add 1 ml paraldehyde. Leave for 24 hours (longer if a pink ring appears when a drop of the solution is spotted onto filter paper). Filter and discard the filtrate. Wash the residue with 50 ml water, then dry the filter paper and its contents in ab oven at 60C. Collect the dry aldehyde fuchsine powder in a little screw-cap bottle. The dye must be pararosaniline (CI 42500). The other components of basic fuchsine are not suitable for this stain. You can use acetaldehyde instead of its cyclic trimer, paraldehyde. Acetaldehyde boils at 21C so it's less convenient to use, but unlike paraldehyde it isn't a controlled drug. Acetaldehyde is formed when paraldehyde reacts with acidified water. Staining solution. Aldehyde fuchsine powder 0.25 g 70% ethanol 200 ml Glacial acetic acid 2.0 ml Leave overnight to dissolve. The solution does not need to be filtered. Method. 1. Stain hydrated paraffin sections, 5 minutes. 2. Rinse in running tap water (messy). 3. Immerse in 95% alcohol with 0.5% conc hydrochloric acid until no more colour is extracted (usually half a minute, but not critical; acid alcohol doesn't remove the real staining). 4. Counterstain if desired (e.g. haemalum and fast green FCF). 5. Wash, dehydrate, clear and coverslip. In some methods (eg for cystine-rich proteins including insulin and classical neurosecretory material) an oxidizing step is put in before staining: 0.5% potassium permanganate in 2% sulphuric acid, 5 min, followed by a rinse in 1% oxalic acid to remove brown deposits of manganese dioxide from the sections, then wash in water. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Marshall Terry Dr, Consultant Histopathologist" wrote: > > John tells us: > "It is also possible to recover > the aldehyde fuchsine as a precipitate that can > be collected by filtering, dried and stored > for 10 or more years." > > OK John ......how? > (This would solve my problem of needing it every 4 months only and suffering the groans of the techs at having to make it up again) > > BTW - I am sure that someone suggested a paraldehyde substitute at one time, but I omitted to keep the post. > > 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: John A. Kiernan [mailto:jkiernan@uwo.ca] > Sent: 17 August 2005 16:11 > To: Histonet > Subject: Re: [Histonet] aldehyde fuchsin > > The solution of aldehyde fuchsine has a rather > short shelf-life (8 weeks) so it's best to make > up your own. It is also possible to recover > the aldehyde fuchsine as a precipitate that can > be collected by filtering, dried and stored > for 10 or more years. Solutions made from aldehyde > fuchsine powder (0.125% in 70% ethanol with > 1% acetic acid) are more stable than the traditional > directly made solution, being good for about 2 years. > > It is said that solutions of aldehyde fuchsine > made from the powder will not stain pancreatic > islet B cells without prior permanganate oxidation > (see Mowry,RW 1978 Stain Technol. 53: 141-154. This > paper also has useful tips for other uses of > aldehyde fuchsine. > -- > ------------------------------- > 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 > _______________________________ > Robin Newlin wrote: > > > > Hi All > > Does anyone know if aldehyde fuchsin is available commercially, or do I have to brew my own. > > Thanks > > Robbin Newlin > > Manager, Cell Imaging/Histology Core Facility > > The Burnham Institute > > 10901 N. Torrey Pines Road > > LaJolla, Ca. 92037 > > 858-646-3100 ext.3552 > > _______________________________________________ > > Histonet mailing list > > Histonet@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 STEGTM <@t> samcstl.org Wed Aug 17 11:36:41 2005 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Slide/block storage Message-ID: Hey out there. We all know we're supposed to keep slides/blocks nearly throughout eternity; question, what are you doing with the archived (read will no longer fit on campus) blocks and slides. We have been using a records etc. storage service, but the retrieval times, and often, chances of accurately getting what we need to retrieve instead of something else, or being told that the items needed cannot be found are becoming more frequent. So...... we're looking for suggestions/alternatives. If you know of a good, reliable storage system, and are in the St. Louis metro area, we'd like to hear from you! We would appreciate all/any replies. Peace, Terre S. HT/ASCP From TJJ <@t> Stowers-Institute.org Wed Aug 17 11:56:09 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Re: Rabbit Anti-sera Background Message-ID: Trey, Perhaps your detection system is TOO sensitive for this antibody. In some cases, rather than take the dilution of the primary antibody out to a ridiculous level, we have opted to use an HRP labeled anti-rabbit secondary if we find the positive staining very intense and background too high. Also, have you tried different antigen unmasking procedures? It could be you don't actually need the power of EDTA to open up the binding sites. In any antibody development, it's best to try an enzyme (trypsin, pronase, protease) digestion in addition to a citrate buffer type formulation, and also keep one set of slides with no pretreatment. If your results are weak or negative, then move on to the more powerful enzymes and high and/or low pH HIER. It could be you're hitting it with much more power than it really needs to stain well. I do really admire your approach to troubleshooting. Well done. Hope this helps, Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From kaburns <@t> med.unc.edu Wed Aug 17 11:57:43 2005 From: kaburns <@t> med.unc.edu (Kim Burns) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] RE: Histonet Digest, Vol 21, Issue 20 In-Reply-To: <200508161703.j7GH3f4p004949@smtp.med.unc.edu> Message-ID: <200508171657.j7HGvhEA005662@smtp.med.unc.edu> Katia, In our lab pregnant people do no coverslipping or other procedures which would put them in contact with xylene or its fumes. Kimberlie Burns C.F Center University of North Carolina Chapel Hill, NC Message: 4 Date: Mon, 15 Aug 2005 18:13:24 -0300 From: "kccatunda" Subject: [Histonet] Headache X pregnancy To: "histonet" Message-ID: Content-Type: text/plain; charset=iso-8859-1 Does anybody has complains of preagnant technicians with headaches because of the smell of reagents such as xylol? What are the precautions you have with preagnancy? Our mounting and staining are mannual procedures but we do have one exaustor for fumes that covers the staining battery. My worry is that we don4t use coverslippers, instead we use a kind of automotive coating (yeah... I know... bad bad bad...). Does somebody has any idea to substitute it without using coverslippers? Thanks again Katia ------------------------------ *************************************** From eric <@t> psl-equip.com Wed Aug 17 12:40:32 2005 From: eric <@t> psl-equip.com (Eric) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Histotech Position in Roseburg, OR Message-ID: Dear Histonetters, One of our clients, Advanced Skin Center, in Roseburg, Oregon, has just lost their tech they use. She was processing Dermatology biopsies with standard H&E stains (No IHC or special stains), and performing frozen section cutting once per week for Mohs surgery. The Dr. has asked us to find a new tech for them. This would be an independent position, one tech performing all functions in the lab. He has no requirement for licensing (it will help though), but would like someone with 1-2 years experience in an histology lab. If anyone is interested, please contact Dr. Paul Reicherter @ (541) 672-7546. Eric Swenson Technical and Customer Service Pacific Southwest Lab Equipment Inc. Phone: (760) 295-1842 Fax: (760) 295-1850 E-mail: Eric@psl-equip.com From ihcatdch <@t> yahoo.com Wed Aug 17 12:44:35 2005 From: ihcatdch <@t> yahoo.com (Phyllis Thaxton) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Richard Allen Coverslipper Message-ID: <20050817174435.75771.qmail@web35204.mail.mud.yahoo.com> Hi HistPeople! Does anyone have any experience with the Richard-Allen Coverslipper or Stainer/Coverslipper Combined automation? Thanks for any feedback! Phyllis Thaxton Roll Tide Roll!!! __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From jwatson <@t> gnf.org Wed Aug 17 14:42:08 2005 From: jwatson <@t> gnf.org (James Watson) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Fluorescent triple staining Message-ID: A coworker is staining with three primary antibodies of the following species: rabbit, goat, and mouse. He wants to use a goat anti-rabbit, rabbit anti-goat, and goat anti-mouse secondary antibodies. The questions are, will we get cross reaction between the secondary antibodies? What would be the recommended blocking solution? I recommended that he use secondary antibodies that are not of the same species of any of the primaries, then we could cocktail the blocking serums, primary antibodies, and secondary antibodies. Any other suggestions? Thank you. James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org From sandosis <@t> uia.net Wed Aug 17 14:45:21 2005 From: sandosis <@t> uia.net (sandosis@uia.net) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Lead HT Position in Laguna Hills, CA Message-ID: <200508171945.j7HJjMr40618@smtp3.uia.net> Full time Lead Histology position, days, Mon-Fri, occasional Saturday. Routine hospital histology work, no IHC. Laguna Hills, CA at Saddleback Memorial Medical Center. --------------------------------------------- This message was sent using the UIA Web Mail Server. ULTIMATE Internet Access, Inc http://www.uia.net/ From pruegg <@t> ihctech.net Wed Aug 17 16:55:16 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Fluorescent triple staining In-Reply-To: Message-ID: <200508172155.j7HLtGfw020594@chip.viawest.net> If you are doing this all on the same section at the same time, I can see that you would have trouble using a secondary that is the same species of one of the primary antibodies. As for blocking, I use a 5-10% solution of non-immune serum from the host of the secondary antibodies and I would make a cocktail of all three. I use this block before the primary and then again before the secondary antibody which results in very specific staining results in my hands. In my experience non-specific staining or background staining is usually unwanted binding of the secondary reagent. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of James Watson Sent: Wednesday, August 17, 2005 12:42 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fluorescent triple staining A coworker is staining with three primary antibodies of the following species: rabbit, goat, and mouse. He wants to use a goat anti-rabbit, rabbit anti-goat, and goat anti-mouse secondary antibodies. The questions are, will we get cross reaction between the secondary antibodies? What would be the recommended blocking solution? I recommended that he use secondary antibodies that are not of the same species of any of the primaries, then we could cocktail the blocking serums, primary antibodies, and secondary antibodies. Any other suggestions? Thank you. James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cjohnson <@t> mail.rockefeller.edu Wed Aug 17 17:04:31 2005 From: cjohnson <@t> mail.rockefeller.edu (Carolyn Johnson) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] antigen retrieval for c-fos or preproenkephalin Message-ID: <200508172204.j7HM4aqM025460@smtp2.rockefeller.edu> Hello, I am doing indirect immunofluorescence on 4% paraformaldehyde fixed (5 hours) mouse tissue, cut to 40 um. I cannot achieve labeling for either c-fos (Chemicon) or pre-proenkephalin (bachem). Has anyone had experience with either of these antibodies? Also, the companies recommended that I try antigen retrieval. If anyone has a protocol that works for either of these antigens, I would greatly appreciate it! Thanks, Carolyn From laurie.reilly <@t> jcu.edu.au Wed Aug 17 18:33:15 2005 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Breast Processing Protocol In-Reply-To: <080566D001A3D9459FFC0A391A646C9106B1B19E@marxchg03.mar.med .navy.mil> Message-ID: <5.1.0.14.0.20050818093003.00c56eb0@mail.jcu.edu.au> Douglas and fellow histonetters, The major problem with processing fatty tissues, assuming that they are fixed properly, is that Ethanol is not a good solvent for fat and therefore connot penetrate the tissues completely, so the tissues are inadequately dehydrated. We have had some success with lipomas by adding a "degreasing" step of xylene into the processing schedule. 70% ethanol 80% ethanol 90% ethanol 95% ethanol Absolute ethanol Xylene Absolute ethanol Xylene Xylene Paraffin Paraffin Paraffin The first Absolute ethanol will dehydrate the tissue to some extent. The next Xylene step will remove most of the fat and then the second Absolute ethanol can complete the dehydration. A compromise situation that we use routinely is to have Absolute ethanol 50:50 Absolute ethanol:Xylene Xylene This is not quite as effective but it is less disruptive to the normal schedule and handles moderately fatty tissues. Regards from Townsville, Australia. Laurie. At 09:58 AM 17/08/2005 -0400, DDDeltour@mar.med.navy.mil wrote: >I am looking for a program for processing fatty breast tissue. We currently >let the breast sit in cassettes in Pen-Fix for an extra day before >processing. We then process the first Fixative in Pen-Fix for one hour then >in 10% Formalin for the second hour (I do not like this but that is they way >it was done when I got here). The next steps are.. 95% one hour X3, 100% one >hour X3, Xylene 1 hour X2. Paraffin for one hour X3. I am considering >replacing one of the 95% with another fixative. What programs do you use for >this? They don't like to hear Sh in Sh out:-). Thank you! > > > >DOUGLAS D. DELTOUR >HISTOLOGY TECHNICIAN >NMC PORTSMOUTH VA > > > >_______________________________________________ >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 Kemlo.Rogerson <@t> elht.nhs.uk Thu Aug 18 02:17:30 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Help with Processing run Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4EE@elht-exch1.xelht.nhs.uk> Good question. . In my youth I 'played' with different clearing agents and solvents. If you were clever and the processing machine had the scope, then you could have 'safe havens' of clearing agents that don't overharden. You could also have a final empty station that allowed the wax to harden. Good answer? Well I tried; it just sounded a good idea yesterday, now not so sure! -----Original Message----- From: Bonnie Whitaker [mailto:bwhitaker@brownpathology.com] Sent: 17 August 2005 15:06 To: Rogerson Kemlo (ELHT) Pathology; 'cindy keith'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Help with Processing run To play the devil's advocate: then what good is altering the processing schedule from home via internet ? Theoretically, changing the protocol to run 3 extra hours, regardless of what step(s) are lengthened, destandardizes the processing schedule. Of course, in reality, there will be times that the schedule is altered. As for how badly does it hurt the tissue to set for 3 additional hours in paraffin, I think that depends on the tissue. If they are large, fatty, mastectomy specimens, they will perhaps even benfit. If they are little GI bx's, then that's another ball of wax. If it is a biopsy run, then I do vote for getting someone to just remove the tissue and set it on the counter. Yes, it will delay it a little while it melts, but if you call in sick, they'll be running behind anyway, so what's a few more minutes. One quicker way to melt them (I think) is to submerse the cassettes in molten paraffin for a few minutes, and then put them in the embedding center (as opposed to just putting them in the embedding center to melt). Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rogerson Kemlo (ELHT) Pathology Sent: Wednesday, August 17, 2005 3:35 AM To: cindy keith; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Help with Processing run Personally, no, but the problem sits with your Operational Procedures. Both they and ICC require standardisation and having differing wax impregnation time's flies in the face of good laboratory practice. Hardening then re-melting is an answer but that will delay blocking out until the wax had melted again. Maybe the answer is to never be sick or have a backup processing cycle that finishes 3 hours later; I concede that will not accommodate your first day of sickness but then there would not be anyone there to take the biopsies out of wax to harden anyway; would there? You need a processing machine connected to the internet or dial up that can be altered remotely; Biochemistry has them! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of cindy keith Sent: 17 August 2005 08:47 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Help with Processing run I started a new job in a lab that has always only had a day shift. My new hours here are 3 am -11am replacing a person who worked 530am-2pm. This change was not well received by my fellow coworkers/supervisor. My question is if the processing run ends at 3am and if I were to be out sick would you think it would be ok to leave bx/lg tissue sitting in the parrafin at 62-64 degrees for 2 1/2 hours-to potentially 3 hours waiting for the next person to come in or would you think it would be better for the tissue to be removed by a fellow lab person and hardened on counter then remelted to embed upon someone else arriving? Immunos are performed on this tissue also. Your responses would be greatly appreciated. _______________________________________________ Histonet mailing list Histonet@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 Thu Aug 18 02:24:52 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Breast Processing Protocol Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD4EF@elht-exch1.xelht.nhs.uk> Sometimes, with fatty tissue, it is useful to go through a fat solvent and then to re-fix. What I used to do was to dehydrate, then go through xylene (which is a fat solvent, then back to 100% alcohol (which is also a fixative) then through xylene to wax. The results were better but I never understood if it was due to the extra processing or the removal of fat. Anyway the logic of it appealed to me. There are other fat solvents you could try. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of DDDeltour@mar.med.navy.mil Sent: 17 August 2005 14:58 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Breast Processing Protocol I am looking for a program for processing fatty breast tissue. We currently let the breast sit in cassettes in Pen-Fix for an extra day before processing. We then process the first Fixative in Pen-Fix for one hour then in 10% Formalin for the second hour (I do not like this but that is they way it was done when I got here). The next steps are.. 95% one hour X3, 100% one hour X3, Xylene 1 hour X2. Paraffin for one hour X3. I am considering replacing one of the 95% with another fixative. What programs do you use for this? They don't like to hear Sh in Sh out:-). Thank you! DOUGLAS D. DELTOUR HISTOLOGY TECHNICIAN NMC PORTSMOUTH VA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Thu Aug 18 02:32:26 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Slide/block storage Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698EC7@elht-exch1.xelht.nhs.uk> Two ways I found of ethical disposal; one let rats eat the blocks, two put records in a 'tank room' and watch it flood. Bingo, job done...... CPA might not like it though, might they? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Therersa Stegall Sent: 17 August 2005 17:37 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide/block storage Hey out there. We all know we're supposed to keep slides/blocks nearly throughout eternity; question, what are you doing with the archived (read will no longer fit on campus) blocks and slides. We have been using a records etc. storage service, but the retrieval times, and often, chances of accurately getting what we need to retrieve instead of something else, or being told that the items needed cannot be found are becoming more frequent. So...... we're looking for suggestions/alternatives. If you know of a good, reliable storage system, and are in the St. Louis metro area, we'd like to hear from you! We would appreciate all/any replies. Peace, Terre S. HT/ASCP _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jefontenot <@t> rota.med.navy.mil Thu Aug 18 02:55:36 2005 From: jefontenot <@t> rota.med.navy.mil (Fontenot, Jason E. HM2) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Slide/block storage Message-ID: According to CAP... Blocks are only required to be kept for a Max of 5 years.... And Slides are to be kept for 10 years.... I dispose of blocks in the biohazard waste with my wet tissue and the slides go in to the Medical (green) glass recycle/Disposal bend or you can gather Sharps containers and fill them up... Neatly of course if you just throw the slides in you will fill it quickly and will not have utilized all the free space... The best way is to keep the lid off neatly stack the old glass slides in a clean unused sharps and when full place the lid on.... The max for my hospital for a sharps container is 50 pounds per sharps container. -----Original Message----- From: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: Thursday, August 18, 2005 9:32 AM To: Therersa Stegall; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Slide/block storage Two ways I found of ethical disposal; one let rats eat the blocks, two put records in a 'tank room' and watch it flood. Bingo, job done...... CPA might not like it though, might they? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Therersa Stegall Sent: 17 August 2005 17:37 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide/block storage Hey out there. We all know we're supposed to keep slides/blocks nearly throughout eternity; question, what are you doing with the archived (read will no longer fit on campus) blocks and slides. We have been using a records etc. storage service, but the retrieval times, and often, chances of accurately getting what we need to retrieve instead of something else, or being told that the items needed cannot be found are becoming more frequent. So...... we're looking for suggestions/alternatives. If you know of a good, reliable storage system, and are in the St. Louis metro area, we'd like to hear from you! We would appreciate all/any replies. Peace, Terre S. 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 jefontenot <@t> rota.med.navy.mil Thu Aug 18 03:08:31 2005 From: jefontenot <@t> rota.med.navy.mil (Fontenot, Jason E. HM2) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Slide/block storage Message-ID: Sorry... I was looking at an old form... Blocks, Slides, paper reports with no computer records or 10 years then dispose.... Accession records and Maintenance records are 2 years... This is the most updated material... -----Original Message----- From: Fontenot, Jason E. HM2 [mailto:jefontenot@rota.med.navy.mil] Sent: Thursday, August 18, 2005 9:56 AM To: 'Rogerson Kemlo (ELHT) Pathology'; Therersa Stegall; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Slide/block storage According to CAP... Blocks are only required to be kept for a Max of 5 years.... And Slides are to be kept for 10 years.... I dispose of blocks in the biohazard waste with my wet tissue and the slides go in to the Medical (green) glass recycle/Disposal bend or you can gather Sharps containers and fill them up... Neatly of course if you just throw the slides in you will fill it quickly and will not have utilized all the free space... The best way is to keep the lid off neatly stack the old glass slides in a clean unused sharps and when full place the lid on.... The max for my hospital for a sharps container is 50 pounds per sharps container. -----Original Message----- From: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: Thursday, August 18, 2005 9:32 AM To: Therersa Stegall; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Slide/block storage Two ways I found of ethical disposal; one let rats eat the blocks, two put records in a 'tank room' and watch it flood. Bingo, job done...... CPA might not like it though, might they? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Therersa Stegall Sent: 17 August 2005 17:37 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide/block storage Hey out there. We all know we're supposed to keep slides/blocks nearly throughout eternity; question, what are you doing with the archived (read will no longer fit on campus) blocks and slides. We have been using a records etc. storage service, but the retrieval times, and often, chances of accurately getting what we need to retrieve instead of something else, or being told that the items needed cannot be found are becoming more frequent. So...... we're looking for suggestions/alternatives. If you know of a good, reliable storage system, and are in the St. Louis metro area, we'd like to hear from you! We would appreciate all/any replies. Peace, Terre S. 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 Terry.Marshall <@t> rothgen.nhs.uk Thu Aug 18 06:25:34 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] aldehyde fuchsin Message-ID: Thanks John. Between you and Gayle, a most useful post concerning one of my tinctorially (if there is such a word) favourite stains. 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: John A. Kiernan [mailto:jkiernan@uwo.ca] Sent: 17 August 2005 17:36 To: Marshall Terry Dr, Consultant Histopathologist Cc: Histonet Subject: Re: [Histonet] aldehyde fuchsin This is the method in M. Gabe's "Histological Techniques" (1976); it's also in my textbook. A similar method was published by Rosa,CC 1953 Stain Technol. 28:299-302. For comparisons of aldehyde fuchsine made in different ways, see papers by GW Nettleton in J. Histochem. Cytochem. 1980-1982. Dissolve 1 g pararosaniline (CI 42500) in 200 ml water. Heat to boiling and leave to cool to room temperature. Add 2.0 ml concentrated hydrochloric acid. Add 1 ml paraldehyde. Leave for 24 hours (longer if a pink ring appears when a drop of the solution is spotted onto filter paper). Filter and discard the filtrate. Wash the residue with 50 ml water, then dry the filter paper and its contents in ab oven at 60C. Collect the dry aldehyde fuchsine powder in a little screw-cap bottle. The dye must be pararosaniline (CI 42500). The other components of basic fuchsine are not suitable for this stain. You can use acetaldehyde instead of its cyclic trimer, paraldehyde. Acetaldehyde boils at 21C so it's less convenient to use, but unlike paraldehyde it isn't a controlled drug. Acetaldehyde is formed when paraldehyde reacts with acidified water. Staining solution. Aldehyde fuchsine powder 0.25 g 70% ethanol 200 ml Glacial acetic acid 2.0 ml Leave overnight to dissolve. The solution does not need to be filtered. Method. 1. Stain hydrated paraffin sections, 5 minutes. 2. Rinse in running tap water (messy). 3. Immerse in 95% alcohol with 0.5% conc hydrochloric acid until no more colour is extracted (usually half a minute, but not critical; acid alcohol doesn't remove the real staining). 4. Counterstain if desired (e.g. haemalum and fast green FCF). 5. Wash, dehydrate, clear and coverslip. In some methods (eg for cystine-rich proteins including insulin and classical neurosecretory material) an oxidizing step is put in before staining: 0.5% potassium permanganate in 2% sulphuric acid, 5 min, followed by a rinse in 1% oxalic acid to remove brown deposits of manganese dioxide from the sections, then wash in water. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Marshall Terry Dr, Consultant Histopathologist" wrote: > > John tells us: > "It is also possible to recover > the aldehyde fuchsine as a precipitate that can > be collected by filtering, dried and stored > for 10 or more years." > > OK John ......how? > (This would solve my problem of needing it every 4 months only and suffering the groans of the techs at having to make it up again) > > BTW - I am sure that someone suggested a paraldehyde substitute at one time, but I omitted to keep the post. > > 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: John A. Kiernan [mailto:jkiernan@uwo.ca] > Sent: 17 August 2005 16:11 > To: Histonet > Subject: Re: [Histonet] aldehyde fuchsin > > The solution of aldehyde fuchsine has a rather > short shelf-life (8 weeks) so it's best to make > up your own. It is also possible to recover > the aldehyde fuchsine as a precipitate that can > be collected by filtering, dried and stored > for 10 or more years. Solutions made from aldehyde > fuchsine powder (0.125% in 70% ethanol with > 1% acetic acid) are more stable than the traditional > directly made solution, being good for about 2 years. > > It is said that solutions of aldehyde fuchsine > made from the powder will not stain pancreatic > islet B cells without prior permanganate oxidation > (see Mowry,RW 1978 Stain Technol. 53: 141-154. This > paper also has useful tips for other uses of > aldehyde fuchsine. > -- > ------------------------------- > 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 > _______________________________ > Robin Newlin wrote: > > > > Hi All > > Does anyone know if aldehyde fuchsin is available commercially, or do I have to brew my own. > > Thanks > > Robbin Newlin > > Manager, Cell Imaging/Histology Core Facility > > The Burnham Institute > > 10901 N. Torrey Pines Road > > LaJolla, Ca. 92037 > > 858-646-3100 ext.3552 > > _______________________________________________ > > Histonet mailing list > > Histonet@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 klapka <@t> uni-duesseldorf.de Thu Aug 18 07:33:11 2005 From: klapka <@t> uni-duesseldorf.de (klapka) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Fluorescence backgound Message-ID: <43048007.8060206@uni-duesseldorf.de> Hello all, does anyone use a method to suppress autofluorescence of tissue due to PFA fixation? I have large background in my PFA fixated nervous tissue... Thank you, Nicole -- Nicole Klapka PhD Labor f?r Molekulare Neurobiologie TVA, Geb. 22.22 Heinrich-Heine Universit?t D?sseldorf Universit?tsstr. 1 40225 D?sseldorf Tel: ++49 211 81 14437 From JWEEMS <@t> sjha.org Thu Aug 18 07:43:56 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Position in Atlanta Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA4640D@sjhaexc02.sjha.org> Saint Joseph's Hospital in Atlanta, GA is seeing an experienced Histotech. Requirements: 2 years of histology experience in a hospital lab HT (ASCP) certification required. If interested, apply at http://www.stjosephsatlanta.org/ 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 pwg1 <@t> cdc.gov Thu Aug 18 07:47:43 2005 From: pwg1 <@t> cdc.gov (Greer, Patricia) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Rabbit Anti-sera Background Message-ID: Trey, You might try blocking with 20% normal sheep serum - we use it routinely in our immuno procedures and it takes care of most of the nonspecific staining. Pat Greer Infectious Disease Pathology Activity Centers for Disease Control and Prevention Mail Stop G-32 Atlanta, GA 30333 404-639-2811 We are having some problems with extensive background and nonspecific staining with a new rabbit polyclonal anti-sera. Here are some of the staining details: -FFPE human tissue -1mM EDTA pH 8.0 HIER for 20 minutes in steamer seems to provide best staining -10% H2O2 block for 10 minutes @ RT (standard for our lab) -block with 1% BSA in TBS for approx. 30 minutes @ RT (no rinse afterwards, just drain off blocking reagents) -primary antibody dilution worked out to 1:1000-2000 (diluted in Dako diluent) for 30 minutes @ RT via titration runs -Dako Envision+ Goat Anti-Rabbit/HRP Labeled polymer detection used for 30 minutes @ RT (standard for our lab) -Dako DAB+ for 10 minutes @ RT (standard for our lab) -counter stained w/ Dako Hematoxylin for 1-2 minutes (standard for our lab) -All rinses done in Dako TBS w/ 0.05% Dako Tween 20 (standard for our lab) We have tried doing peptide blocking experiments that have shown some of the staining can be blocked, however most of the staining is still present, suggesting that it is due to nonspecific binding. We know from Westerns that our blocking experiments include a VERY large excess of peptide, so all of the specific antibodies should be bound to peptide. Negative controls (without antibody and/or with peptide only) always show up DEAD negative. No avidin/biotin to worry about since this detections system is a labeled polymer. The detection system has been working very well for us in the past, including with other commercially available rabbit polyclonals, so I can't imagine this would be the cause of our problem. What suggestions might you have to reduce this intense background/nonspecific staining. 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Thu Aug 18 09:27:13 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] aldehyde fuchsin In-Reply-To: References: Message-ID: <43049AC1.4020606@umdnj.edu> "Staining properties of aldehyde fuchsin analogues" by T.S. Buehner, G.S. Nettleton and J.B. Longley. 1979. J. Histochem. Cytochem. 27(3):782-787. This paper discusses their results with paraldehyde, acetaldehyde, etc. Also, it is important that you prepare the aldehyde fuchsin from pararosanalin not just any old basic fuchsine (see Mowry et al., Stain Technology 55(2):91-103, 1980. And, the shelf life is short ......... One can also make aldehyde thionin ......... Geoff Marshall Terry Dr, Consultant Histopathologist wrote: >Thanks John. >Between you and Gayle, a most useful post concerning one of my tinctorially (if there is such a word) favourite stains. > >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: John A. Kiernan [mailto:jkiernan@uwo.ca] >Sent: 17 August 2005 17:36 >To: Marshall Terry Dr, Consultant Histopathologist >Cc: Histonet >Subject: Re: [Histonet] aldehyde fuchsin > > >This is the method in M. Gabe's "Histological Techniques" >(1976); it's also in my textbook. A similar method >was published by Rosa,CC 1953 Stain Technol. 28:299-302. >For comparisons of aldehyde fuchsine made in different >ways, see papers by GW Nettleton in J. Histochem. >Cytochem. 1980-1982. > >Dissolve 1 g pararosaniline (CI 42500) in 200 ml water. >Heat to boiling and leave to cool to room temperature. >Add 2.0 ml concentrated hydrochloric acid. >Add 1 ml paraldehyde. >Leave for 24 hours (longer if a pink ring >appears when a drop of the solution is spotted >onto filter paper). >Filter and discard the filtrate. >Wash the residue with 50 ml water, then >dry the filter paper and its contents in >ab oven at 60C. Collect the dry aldehyde >fuchsine powder in a little screw-cap >bottle. > >The dye must be pararosaniline (CI 42500). >The other components of basic fuchsine are >not suitable for this stain. You can use >acetaldehyde instead of its cyclic trimer, >paraldehyde. Acetaldehyde boils at 21C so >it's less convenient to use, but unlike >paraldehyde it isn't a controlled drug. >Acetaldehyde is formed when paraldehyde >reacts with acidified water. > >Staining solution. > >Aldehyde fuchsine powder 0.25 g >70% ethanol 200 ml >Glacial acetic acid 2.0 ml > >Leave overnight to dissolve. The >solution does not need to be filtered. > >Method. > >1. Stain hydrated paraffin sections, 5 minutes. >2. Rinse in running tap water (messy). >3. Immerse in 95% alcohol with 0.5% conc >hydrochloric acid until no more colour >is extracted (usually half a minute, but >not critical; acid alcohol doesn't >remove the real staining). >4. Counterstain if desired (e.g. haemalum >and fast green FCF). >5. Wash, dehydrate, clear and coverslip. > >In some methods (eg for cystine-rich >proteins including insulin and classical >neurosecretory material) an oxidizing step >is put in before staining: 0.5% potassium >permanganate in 2% sulphuric acid, 5 min, >followed by a rinse in 1% oxalic acid to >remove brown deposits of manganese dioxide >from the sections, then wash in water. > > -- -- ********************************************** 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 RJLevier <@t> LancasterGeneral.org Thu Aug 18 10:24:07 2005 From: RJLevier <@t> LancasterGeneral.org (LeVier, Rebecca J) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Xylene Substitute ? Message-ID: <0FDBF29200C637468CCC1F9E7E85A3D214E88F@MAIL-LR.lha.org> Hello Everyone, I have a question regarding the Xylene Substitute, Xylene Substitute 2 from Shandon. What is everyone's opinion of the substitute and have you seen anything that could be wrong with using this in every step that Xylene is usually used? Thanks, Becky From gcallis <@t> montana.edu Thu Aug 18 11:18:54 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Rabbit Anti-sera Background In-Reply-To: References: Message-ID: <6.0.0.22.1.20050818100908.01b2b1f8@gemini.msu.montana.edu> Swine serum is also supposed to be excellent. At 06:47 AM 8/18/2005, you wrote: >Trey, > >You might try blocking with 20% normal sheep serum - we use it routinely >in our immuno procedures and it takes care of most of the nonspecific >staining. > >Pat Greer >Infectious Disease Pathology Activity >Centers for Disease Control and Prevention >Mail Stop G-32 >Atlanta, GA 30333 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 Thu Aug 18 11:21:11 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] pararosaniline for aldehyde fuchsin In-Reply-To: <43049AC1.4020606@umdnj.edu> References: <43049AC1.4020606@umdnj.edu> Message-ID: <6.0.0.22.1.20050818101953.01b2a520@gemini.msu.montana.edu> Sigma has an excellent pararosaniline for this purpose At 08:27 AM 8/18/2005, you wrote: > "Staining properties of aldehyde fuchsin analogues" by T.S. Buehner, > G.S. Nettleton and J.B. Longley. 1979. J. Histochem. Cytochem. > 27(3):782-787. This paper discusses their results with paraldehyde, > acetaldehyde, etc. Also, it is important that you prepare the aldehyde > fuchsin from pararosanalin not just any old basic fuchsine (see Mowry et > al., Stain Technology 55(2):91-103, 1980. And, the shelf life is short > ......... > One can also make aldehyde thionin ......... > >Geoff > > >> >>This is the method in M. Gabe's "Histological Techniques" >>(1976); it's also in my textbook. A similar method >>was published by Rosa,CC 1953 Stain Technol. 28:299-302. >>For comparisons of aldehyde fuchsine made in different ways, see papers >>by GW Nettleton in J. Histochem. >>Cytochem. 1980-1982. >> >>Dissolve 1 g pararosaniline (CI 42500) in 200 ml water. Heat to boiling >>and leave to cool to room temperature. >>Add 2.0 ml concentrated hydrochloric acid. >>Add 1 ml paraldehyde. >>Leave for 24 hours (longer if a pink ring >>appears when a drop of the solution is spotted onto filter paper). >>Filter and discard the filtrate. >>Wash the residue with 50 ml water, then >>dry the filter paper and its contents in >>ab oven at 60C. Collect the dry aldehyde >>fuchsine powder in a little screw-cap >>bottle. >> >>The dye must be pararosaniline (CI 42500). >>The other components of basic fuchsine are >>not suitable for this stain. You can use >>acetaldehyde instead of its cyclic trimer, paraldehyde. Acetaldehyde >>boils at 21C so >>it's less convenient to use, but unlike >>paraldehyde it isn't a controlled drug. >>Acetaldehyde is formed when paraldehyde >>reacts with acidified water. >> >>Staining solution. >> >>Aldehyde fuchsine powder 0.25 g >>70% ethanol 200 ml >>Glacial acetic acid 2.0 ml >> >>Leave overnight to dissolve. The >>solution does not need to be filtered. >> >>Method. >> >>1. Stain hydrated paraffin sections, 5 minutes. >>2. Rinse in running tap water (messy). >>3. Immerse in 95% alcohol with 0.5% conc >>hydrochloric acid until no more colour >>is extracted (usually half a minute, but >>not critical; acid alcohol doesn't >>remove the real staining). >>4. Counterstain if desired (e.g. haemalum >>and fast green FCF). >>5. Wash, dehydrate, clear and coverslip. >> >>In some methods (eg for cystine-rich proteins including insulin and classical >>neurosecretory material) an oxidizing step >>is put in before staining: 0.5% potassium >>permanganate in 2% sulphuric acid, 5 min, >>followed by a rinse in 1% oxalic acid to >>remove brown deposits of manganese dioxide >>from the sections, then wash in water. >> > > >-- >-- >********************************************** >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 David.Edmondson <@t> christie-tr.nwest.nhs.uk Thu Aug 18 11:29:00 2005 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Breast Processing Protocol Message-ID: <3C83687E8F6AE04792E361ABE2D385B8418012@cht-mail2-2k.xchristie.nhs.uk> Hi all, We have been using Carnoy's fixative as an intermediate / degreasing stage. Cassettes in a pot of it for a few hours, it would be better on an old processing machine "Dunking" but space does not allow. We rinse the cassettes before processing so we do not get the full benefit of any dehydration that may have occured. Dave, Christie Hospital Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Laurie Reilly Sent: 18 August 2005 00:33 To: DDDeltour@mar.med.navy.mil; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Breast Processing Protocol Douglas and fellow histonetters, The major problem with processing fatty tissues, assuming that they are fixed properly, is that Ethanol is not a good solvent for fat and therefore connot penetrate the tissues completely, so the tissues are inadequately dehydrated. We have had some success with lipomas by adding a "degreasing" step of xylene into the processing schedule. 70% ethanol 80% ethanol 90% ethanol 95% ethanol Absolute ethanol Xylene Absolute ethanol Xylene Xylene Paraffin Paraffin Paraffin The first Absolute ethanol will dehydrate the tissue to some extent. The next Xylene step will remove most of the fat and then the second Absolute ethanol can complete the dehydration. A compromise situation that we use routinely is to have Absolute ethanol 50:50 Absolute ethanol:Xylene Xylene This is not quite as effective but it is less disruptive to the normal schedule and handles moderately fatty tissues. Regards from Townsville, Australia. Laurie. At 09:58 AM 17/08/2005 -0400, DDDeltour@mar.med.navy.mil wrote: >I am looking for a program for processing fatty breast tissue. We currently >let the breast sit in cassettes in Pen-Fix for an extra day before >processing. We then process the first Fixative in Pen-Fix for one hour then >in 10% Formalin for the second hour (I do not like this but that is they way >it was done when I got here). The next steps are.. 95% one hour X3, 100% one >hour X3, Xylene 1 hour X2. Paraffin for one hour X3. I am considering >replacing one of the 95% with another fixative. What programs do you use for >this? They don't like to hear Sh in Sh out:-). Thank you! > > > >DOUGLAS D. DELTOUR >HISTOLOGY TECHNICIAN >NMC PORTSMOUTH VA > > > >_______________________________________________ >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 ****************************************************************************************************************** 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 Kristopher.Kalleberg <@t> unilever.com Thu Aug 18 12:19:29 2005 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] paraffin problem Message-ID: Does anyone have any suggestions on how to get paraffin off of clothes. I recently bought a pair of corduroy pants and spilled paraffin down them while in the lab and know of no way to get the paraffin out. Suggestions would be greatly appreciated. Thanks. Kris Kalleberg Histologist Unilever R&D 40 Merritt Blvd. Trumbull, CT 06611 203 381 5765 From PMonfils <@t> Lifespan.org Thu Aug 18 12:20:51 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Fluorescence backgound Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175B2@lsexch.lsmaster.lifespan.org> Autofluorescence in CNS tissue, as well as some other tissues, is often due to the presence of lipofuscin. Staining with Sudan Black B after immunolabeling is often effective in suppressing such background autofluorescence. Make up 0.l% solution in 70% ethanol. Heat to boiling, then cool and filter. Optimum staining time may vary somewhat from one tissue to another and for sections of different thickness, but 10 minutes usually provides an acceptable level of suppression, and often complete suppression of autofluorescence. Quickly rinse off the stain with water or buffer, several rapid changes, and coverslip as usual. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > klapka > Sent: Thursday, August 18, 2005 5:33 AM > To: Histonet > Subject: [Histonet] Fluorescence backgound > > Hello all, > > does anyone use a method to suppress autofluorescence of tissue due to > PFA fixation? I have large background in my PFA fixated nervous tissue... > Thank you, > Nicole > > -- > Nicole Klapka PhD > Labor f?r Molekulare Neurobiologie > TVA, Geb. 22.22 > Heinrich-Heine Universit?t D?sseldorf > Universit?tsstr. 1 > 40225 D?sseldorf > Tel: ++49 211 81 14437 > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From kbroomal <@t> NEMOURS.ORG Thu Aug 18 12:35:28 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] paraffin problem Message-ID: <6E41111281623B4B8A9AB8F9A7EA34378243E0@wlmmsx02.nemours.org> This works with candle wax on fabric: place plain brown paper (like from a paper bag) over the area & apply a hot iron to it. The wax sticks to the paper & comes off the fabric. (also worked once with regular paper for me). Good Luck! Kristen Broomall -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kristopher Kalleberg Sent: Thursday, August 18, 2005 1:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] paraffin problem Does anyone have any suggestions on how to get paraffin off of clothes. I recently bought a pair of corduroy pants and spilled paraffin down them while in the lab and know of no way to get the paraffin out. Suggestions would be greatly appreciated. Thanks. Kris Kalleberg Histologist Unilever R&D 40 Merritt Blvd. Trumbull, CT 06611 203 381 5765 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From timothy.macatee <@t> med.nyu.edu Thu Aug 18 12:33:55 2005 From: timothy.macatee <@t> med.nyu.edu (Timothy Macatee) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] paraffin problem In-Reply-To: Message-ID: I saw this on The Early Show on CBS. They did it on carpeting, but it should work on clothes. Remove all crusted paraffin then place a paper towel over the remaining paraffin and soak up the wax using a warm iron. It usually takes several paper towels but it does work. Tim From king.laurie <@t> marshfieldclinic.org Thu Aug 18 12:37:53 2005 From: king.laurie <@t> marshfieldclinic.org (king.laurie@marshfieldclinic.org) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] paraffin problem Message-ID: <606f01c5a41b$8fdf8570$8e0110ac@mfldclinframe.org> Kriss, I used to have some success getting paraffin out of clothing by ironing it face down onto a brown paper bag-you are then left with a grease stain on the bag and residual on the cloth. At this point, a good laundry degreaser like Shout gel rubbed in and let set a couple of times before laundering. Don't put them in a hot dryer until you see that it is gone. The dryer will just set it in for good. Good luck. Laurie King ------Original Message------ From: "Kristopher Kalleberg" Date: Thu Aug 18, 2005 -- 12:23:05 PM To: "histonet@lists.utsouthwestern.edu" Subject: [Histonet] paraffin problem Does anyone have any suggestions on how to get paraffin off of clothes. I recently bought a pair of corduroy pants and spilled paraffin down them while in the lab and know of no way to get the paraffin out. Suggestions would be greatly appreciated. Thanks. Kris Kalleberg Histologist Unilever R&D 40 Merritt Blvd. Trumbull, CT 06611 203 381 5765 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet >From histonet-bounces@lists.utsouthwestern.edu Thu Aug 18 12:22:42 2005 From gcallis <@t> montana.edu Thu Aug 18 12:37:58 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Fluorescence backgound In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE017175B2@lsexch.lsmaster.l ifespan.org> References: <09C945920A6B654199F7A58A1D7D1FDE017175B2@lsexch.lsmaster.lifespan.org> Message-ID: <6.0.0.22.1.20050818113554.01b24e20@gemini.msu.montana.edu> I think the Chemicon autofluorescence removal reagent - whatever they call it, is to get rid of lipofuscins - their website will have the information how it works. At 11:20 AM 8/18/2005, you wrote: >Autofluorescence in CNS tissue, as well as some other tissues, is often due >to the presence of lipofuscin. Staining with Sudan Black B after >immunolabeling is often effective in suppressing such background >autofluorescence. > >Make up 0.l% solution in 70% ethanol. Heat to boiling, then cool and >filter. Optimum staining time may vary somewhat from one tissue to another >and for sections of different thickness, but 10 minutes usually provides an >acceptable level of suppression, and often complete suppression of >autofluorescence. Quickly rinse off the stain with water or buffer, several >rapid changes, and coverslip as usual. > >Paul M. > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > > klapka > > Sent: Thursday, August 18, 2005 5:33 AM > > To: Histonet > > Subject: [Histonet] Fluorescence backgound > > > > Hello all, > > > > does anyone use a method to suppress autofluorescence of tissue due to > > PFA fixation? I have large background in my PFA fixated nervous tissue... > > Thank you, > > Nicole > > > > -- > > Nicole Klapka PhD > > Labor f?r Molekulare Neurobiologie > > TVA, Geb. 22.22 > > Heinrich-Heine Universit?t D?sseldorf > > Universit?tsstr. 1 > > 40225 D?sseldorf > > Tel: ++49 211 81 14437 > > > > > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From PMonfils <@t> Lifespan.org Thu Aug 18 12:39:07 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] paraffin problem Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175B3@lsexch.lsmaster.lifespan.org> You mean you don't have a little bottle of xylene on the shelf next to your washing machine? :-) I do! I use it to spot-treat paraffin spills and sometimes other types of non-water soluble stains like oil or grease. Most common clearing agents can be applied to most fabrics without harming either the fabric or the dyes in the fabric. I usually apply the xylene with a small plastic dropper, out on the patio, which is right off the laundry area. I wait about 10 minutes after applying the solvent, then just drop the item into the washer while it is already washing (water as hot as the garment will allow, and double the usual amount of detergent). Both the clearing agent and the dissolved paraffin wash out quickly. I did have one little problem with one of my wife's blouses though. She spilled candle wax on it. The blouse was white with little pink and blue flowers, and green leaves. I brought it into work, and I applied chloroform to the spot. It removed the wax quickly. And the blue flowers. Oops. Paul M. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Kristopher Kalleberg > Sent: Thursday, August 18, 2005 10:19 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] paraffin problem > > Does anyone have any suggestions on how to get paraffin off of clothes. I > recently bought a pair of corduroy pants and spilled paraffin down them > while > in the lab and know of no way to get the paraffin out. Suggestions would > be > greatly appreciated. Thanks. > > Kris Kalleberg > Histologist > Unilever R&D > 40 Merritt Blvd. > Trumbull, CT 06611 > 203 381 5765 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From algranth <@t> u.arizona.edu Thu Aug 18 13:11:33 2005 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] paraffin problem In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE017175B3@lsexch.lsmaster.l ifespan.org> Message-ID: <4.3.2.7.2.20050818110830.00cc78d8@algranth.inbox.email.arizona.edu> Goo-Gone works too. You can find this at places like Walmart, Bed, Bath and Beyond, etc. I have it in my laundry room and use it for paraffin related spots and some other things too. It works well when your kid washes/dries his clothes and doesn't take the chapstik out of a pocket. Andi At 01:39 PM 8/18/2005 -0400, Monfils, Paul wrote: >You mean you don't have a little bottle of xylene on the shelf next to your >washing machine? :-) I do! I use it to spot-treat paraffin spills and >sometimes other types of non-water soluble stains like oil or grease. Most >common clearing agents can be applied to most fabrics without harming either >the fabric or the dyes in the fabric. I usually apply the xylene with a >small plastic dropper, out on the patio, which is right off the laundry >area. I wait about 10 minutes after applying the solvent, then just drop the >item into the washer while it is already washing (water as hot as the >garment will allow, and double the usual amount of detergent). Both the >clearing agent and the dissolved paraffin wash out quickly. > >I did have one little problem with one of my wife's blouses though. She >spilled candle wax on it. The blouse was white with little pink and blue >flowers, and green leaves. I brought it into work, and I applied chloroform >to the spot. It removed the wax quickly. And the blue flowers. Oops. > >Paul M. > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > > Kristopher Kalleberg > > Sent: Thursday, August 18, 2005 10:19 AM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] paraffin problem > > > > Does anyone have any suggestions on how to get paraffin off of clothes. I > > recently bought a pair of corduroy pants and spilled paraffin down them > > while > > in the lab and know of no way to get the paraffin out. Suggestions would > > be > > greatly appreciated. Thanks. > > > > Kris Kalleberg > > Histologist > > Unilever R&D > > 40 Merritt Blvd. > > Trumbull, CT 06611 > > 203 381 5765 > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From anh2006 <@t> med.cornell.edu Thu Aug 18 13:16:04 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] paraffin problem In-Reply-To: References: Message-ID: Try ironing it with a uncoated brown paper bag (grocery store bags work great) over the affected area. Lifting and replacing every few seconds. The wax should melt and be absorbed into the brown paper bag. I do this to remove household candle wax and it works well. >Does anyone have any suggestions on how to get paraffin off of clothes. I >recently bought a pair of corduroy pants and spilled paraffin down them while >in the lab and know of no way to get the paraffin out. Suggestions would be >greatly appreciated. Thanks. > >Kris Kalleberg >Histologist >Unilever R&D >40 Merritt Blvd. >Trumbull, CT 06611 >203 381 5765 >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- From jennifer.l.hofecker <@t> Vanderbilt.Edu Thu Aug 18 13:59:34 2005 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Ahh, my trichromes are green again Message-ID: <898D946569A27444B65667A49C07405207546E@mailbe06.mc.vanderbilt.edu> Hi everyone, Just wanted to thank the many people who gave me helpful hints. We found that our commercially prepared Gomori trichrome solution was at pH 2.0! We brought it up to 3.4 and it worked great. We have ordered new solution and I plan on checking the pH when it comes in. We also decided to wait a few minutes after cutting frozen sections. Several people recommended both of these things and we are very grateful. Have a great rest of the week. Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax From nmuvarak <@t> facstaff.wisc.edu Thu Aug 18 14:12:49 2005 From: nmuvarak <@t> facstaff.wisc.edu (NIDAL EMILIO MUVARAK) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Does anyone know? Message-ID: <187893e1877374.1877374187893e@wiscmail.wisc.edu> How do I unsubscribe? From essssy <@t> gmail.com Thu Aug 18 14:13:39 2005 From: essssy <@t> gmail.com (DrEssy) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Ethanol or acetone fixation Message-ID: <85b7376505081812134445114d@mail.gmail.com> Hi, I was wondering if anyone knows how acetone and/or alcohol fixation works? Does it work in a similar manner as water soluble fixatives such as formalin? Thanks! -Essy From mcauliff <@t> umdnj.edu Thu Aug 18 14:13:40 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Ahh, my trichromes are green again In-Reply-To: <898D946569A27444B65667A49C07405207546E@mailbe06.mc.vanderbilt.edu> References: <898D946569A27444B65667A49C07405207546E@mailbe06.mc.vanderbilt.edu> Message-ID: <4304DDE4.5010100@umdnj.edu> This is a good argument for making your own solutions. That way you will know exactly what is in them and what the pH is. Geoff Hofecker, Jennifer L wrote: >Hi everyone, >Just wanted to thank the many people who gave me helpful hints. We found that our commercially prepared Gomori trichrome solution was at pH 2.0! We brought it up to 3.4 and it worked great. We have ordered new solution and I plan on checking the pH when it comes in. We also decided to wait a few minutes after cutting frozen sections. Several people recommended both of these things and we are very grateful. >Have a great rest of the week. > >Jennifer > > > >Jennifer Hofecker, HT (ASCP) >Vanderbilt University Medical Center >Division of Neuropathology >(615) 343-0083 >(615) 343-7089 fax >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From amoklebu <@t> seattlecca.org Thu Aug 18 14:15:46 2005 From: amoklebu <@t> seattlecca.org (Moklebust, Amanda C) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] CD31 Antibody from Zymed Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B948071D6976@wala01.seattlecca.org> Dear Histonetters, One of our pathologists received an advertisement packet from Zymed Laboratories. He is interested in anyone's experience with their antibody for CD31 (PECAM-1) Clone 1A10. Thank you in advance for your help. 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 jkiernan <@t> uwo.ca Thu Aug 18 14:46:06 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Ethanol or acetone fixation References: <85b7376505081812134445114d@mail.gmail.com> Message-ID: <4304E57E.30C81685@uwo.ca> No, the mechanisms are quite different. Alcohol, acetone and formaldehyde are all water-soluble fixatives. Formaldehyde attaches to protein molecules and (given some time) links them together. Alcohol or acetone coagulates protein, changing the shapes of the molecules but not their chemical properties. Coagulant fixatives do their work instantly as they penetrate the tissue. Formaldehyde (a smaller molecule than ethanol or acetone) penetrates rapidly but needs 12-24 hours to stabilize the structure. Mixtures containing both alcohol and formaldehyde can provide the advantages of both types of fixation. Such mixtures usually contain some acetic acid as well, which coagulates nuclear chromatin and opposes the shrinking action of the alcohol. See http://publish.uwo.ca/~jkiernan/formglut.htm for more about how formaldehyde works. -- ------------------------------- 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 _______________________________ DrEssy wrote: > > Hi, > I was wondering if anyone knows how acetone and/or alcohol fixation works? > Does it work in a similar manner as water soluble fixatives such as > formalin? > Thanks! > -Essy > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Thu Aug 18 15:27:04 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] PU1, BOB1, OCT2 antibodies Message-ID: Hi histonetters, We've been asked to send slides somewhere to get these 3 antibodies done. Are these clones and are they known by any other name? Does anyone out there (reference labs) offer these (preferably without interpretation)? Thanks, Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 From DALEY_C <@t> palmer.edu Thu Aug 18 16:12:12 2005 From: DALEY_C <@t> palmer.edu (Clover Daley) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Fixative for storage of perfused rat organs Message-ID: I am putting a tissue bank together for research students. We are currently perfusing with 2% paraformaldehyde and 1% glutaraldehyde in phosphate buffer to collect tissues for EM studies. The liver, kidney, stomach, brain will be stored. What fixative and temperature of solution would be best. These tissues will be used for light microscopy studies. Also we will be perfusing with 4% paraformaldehyde in phosphate buffer, harvesting the same tissues. Should they be stored in the same solution and do they need to be kept in the refrigerator? Thank you for your time, Clover Daley Palmer Center for Chiropractic Research From anh2006 <@t> med.cornell.edu Thu Aug 18 17:23:50 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Fluorescence backgound In-Reply-To: <6.0.0.22.1.20050818113554.01b24e20@gemini.msu.montana.edu> References: <09C945920A6B654199F7A58A1D7D1FDE017175B2@lsexch.lsmaster.lifespan.org> <6.0.0.22.1.20050818113554.01b24e20@gemini.msu.montana.edu> Message-ID: Has anyone used this? Have you used this yourself Gayle? It worries me that the instructions say to use it after staining and then subsequently do 70% ethanol rinses! Any info provided is most appreciated. Andrea At 11:37 AM -0600 8/18/05, Gayle Callis wrote: >I think the Chemicon autofluorescence removal reagent - whatever >they call it, is to get rid of lipofuscins - their website will have >the information how it works. -- From vitha <@t> mic.tamu.edu Thu Aug 18 17:34:20 2005 From: vitha <@t> mic.tamu.edu (Stanislav Vitha) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] (no subject) Message-ID: <6.2.3.4.0.20050818171504.03e63630@mic.tamu.edu> I have used a setup similar to the one described in the previous reply by Dr. Chiovetti, using a coolpix 4500 camera. You can set a zoom of the coolpix objective to a precise numerical value using the "photopc" freely downloadable software and a serial cable (not the USB cable supplied with the camera; Coolpix serial cable can be purchased for ` $25). There are DOS, windows and Linux versions of the software. see http://www.lightner.net/lightner/bruce/photopc/ I set up an old computer as a linux system and wrote a simple shell script that would on startup set the zoom of the camera to a certain default value (say 12.7 mm), as well as set some other parameters. The computer also serves as a remote release, so that you do not have to spend another $80 for the remote release gadget and just press the "Enter " key to take a picture. If you are interested, I will be happy to send you the shell script and more info on 'photopc" software directly as an attachment. Stan >Message: 8 >Date: Mon, 15 Aug 2005 11:38:55 EDT >From: RCHIOVETTI@aol.com >Subject: Re: [Histonet] Nikon relay lens >To: nyilmaz@mersin.edu.tr, histonet@lists.utsouthwestern.edu >Message-ID: <1c3.2eb8ab83.3032110f@aol.com> >Content-Type: text/plain; charset="US-ASCII" > >In a message dated 8/14/2005 11:35:35 PM US Mountain Standard Time, >nyilmaz@mersin.edu.tr writes: > > > We're using an Olympus BX 50 light microscope attached with Nikon Coolpix > > 5000 digital camera. I'm wondering what the mangnification value > of MDC relay > > lens between the camera and the microscope is. >4. Since the zoom on the camera is infinitely adjustable, we had to decide >on a reproducible zoom setting that always gave the same magnification. For >the Coolpix 8800 and our adapter, we opted to zoom to the maximum >setting. It's >10x on the 8800; I'm not sure what it is on the 5000. > >5. To make measurements we set the camera to this maximum (10x) zoom, and >took pictures of the stage micrometer with each objective. From there, it's >relatively easy to measure the spacing of the 10 micrometer lines and to >calculate a total magnification. > >This was the only way we could come up with to make measurements, since the >camera's zoom is so variable. It's a small inconvenience, but the >camera works >great with this set-up. > >You could probably use a similar procedure, provided you are using the >camera's optical zoom to fill the field of view with the MDC relay >lens. The trick >is to use a zoom setting that is reproducible. > >Hope this is of some help. > >Best regards, > >Robert (Bob) Chiovetti, Ph.D. >The Microscope Works >Arizona's Microscopy Resource >132 North Elster Drive >Tucson, AZ 85710-3212 USA >Tel./Fax 520-546-4986 >Member, Arizona Small Business Association - ASBA Dr. Stanislav Vitha vitha@mic.tamu.edu Microscopy and Imaging Center Texas A&M University BSBW 119 College Station, TX 77843-2257 tel: 979-845-1129 (main desk) tel: 979-845-1607 (direct link) fax: 979-847-8933 From Lynn.Scott <@t> ahss.org Fri Aug 19 08:44:07 2005 From: Lynn.Scott <@t> ahss.org (Scott, Lynn M.) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Solvent Recyclers Message-ID: Looking for comments concerning gravity flow solvent recyclers ? Lynn M. Scott Regional Supervisor Pathology Adventist Lab Partners Hinsdale Hospital 630-856-7859 Lynn.Scott@AHSS.org ------------------------------- 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, 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 djohnson14 <@t> hotmail.com Fri Aug 19 09:34:56 2005 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Mohs coverslippign air bubbles Message-ID: Question: I have mohs tech in OH that is having a lot of problems with air bubbles when coverslipping. THey coverslip by hand and they have tried various mouting medias, etc and still are baffled Any suggestions Dave Johnson From gagnone <@t> KGH.KARI.NET Fri Aug 19 09:52:51 2005 From: gagnone <@t> KGH.KARI.NET (Gagnon, Eric) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Ventana HER2 4B5 Message-ID: Hello Histonetters, Anyone have any experience with Ventana HER2 4B5 antibody? We are running the Ventana XT, and had some pathologist rumblings about c-erbB-2 (ASR)CB11, our previous antibody, giving some staining in non-neoplastic cells. We are switching over to the 4B5, and wondering, can we expect better results? Any feedback appreciated! Eric Gagnon, MLT Kingston General Hospital, Kingston, Ontario From kjnpeppa <@t> yahoo.com Fri Aug 19 09:54:53 2005 From: kjnpeppa <@t> yahoo.com (Barbara Rouse) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Pap staining question Message-ID: <20050819145453.41708.qmail@web51704.mail.yahoo.com> Hello to all: My pathologist states that all of our non-gyn preparations are not staining with any green coloring. We use OG6 and EA36 and have for the last 10 or more years. Any ideas? Thanks Barbara __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From petepath <@t> yahoo.com Fri Aug 19 10:12:14 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Mohs coverslippign air bubbles Message-ID: <20050819151214.99931.qmail@web30409.mail.mud.yahoo.com> Hi Dave, A histotech taught me this technique when I was a resident. Lay down a cover slip on a paper towel with the long axis parrallel to your arm. Put a drop of mounting medium on the coverslip. When my slide comes out of xylene, drip a drop of xylene on the drop of medium to dilute it a bit. Line the slide up on edge next to the cover slip and lever it down so the drop of medium just touches the slide and then lever it back up. Do not just drop the slide on the cover slip. The cover slip will adhere to the slide by cohesive forces and the medium will spread by capillary action leaving little or no bubbles. You can get very fast at this, lay out a bunch of coverslips and cover a number of slides very quickly. Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From gcallis <@t> montana.edu Fri Aug 19 10:23:36 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Fluorescence backgound In-Reply-To: References: <09C945920A6B654199F7A58A1D7D1FDE017175B2@lsexch.lsmaster.lifespan.org> <6.0.0.22.1.20050818113554.01b24e20@gemini.msu.montana.edu> Message-ID: <6.0.0.22.1.20050819085908.01b0ece8@gemini.msu.montana.edu> Andrea, I don't need to use the Chemicon reagent for my work, but it new product being marketed for autofluorescence reduction and something I had not seen before I might try it IF I thought I was having troubles with lipofuscin - the reason I brought it up as someone in a previous message was pointing out lipofuscin creates autofluorescence problems in brain. It was only a suggestion, not based on me using it - however the info has been filed away for future reference. My autofluorescence problems are minimal since we avoid aldehyde fixatives. We prefer all IFA staining on FS fixed with acetone/alcohol mixture or 4C acteone. However, these fixatives will damage DsRed or eGFP (we experience loss of fluorescence with these glowing proteins after solvent fixation) so the new gig is to leave FS unfixed, air dry for 30 min - 1 hr, then coverslip these unfixed, dry FS with Molecular Probes Prolong Gold antifade with DAPI, this is hard set, ready to use. We did not need IFA staining, just trying to see where the DsRed-protein was binding certain cells, and the DAPI was a bonus - morphology wise. The results were wonderful. So many ways ------- I just ran across information from Molecular Probes website this week, part of the Q&A from a "Dr. Tech". If anyone knows about effects of solvents on fluorophores, they are probably the experts - maybe their tech services know more about the Chemicon Autofluorescence "buster"! used with their fluorescent compounds. Q: Will the solvents used for fixing, permeabilizing, or deparaffinizing samples degrade the fluorescent dyes on my conjugated cells, antibodies, proteins, etc.? How can I avoid degrading my fluorescent labels? A: Many chemicals commonly used for fixing, permeabilizing, or deparaffinizing samples will not degrade fluorescent dyes conjugated to your biomolecules and cells. Formaldehyde, glutaraldehyde, formalin, detergents (e.g., Triton X-100, saponin), surfactants (e.g., Tween, Brij), alcohols, acetone, and xylene do not promote the decomposition of the dyes. Note that xylene will quench many dyes, but once the xylene is removed, the dyes can still fluoresce. Covalently attached dyes are stable in the presence of all of these reagents. Fluorescent dyes are degraded by extremes of pH (pH <3 and >9 to 12, depending on the dye) and strong oxidizing agents. You may wish to avoid heavy-metal fixation and picric acid. Many heavy metals can either quench fluorescence or promote dye degradation by redox reactions. At 04:23 PM 8/18/2005, you wrote: >Has anyone used this? Have you used this yourself Gayle? It worries me >that the instructions say to use it after staining and then subsequently >do 70% ethanol rinses! > >Any info provided is most appreciated. >Andrea > >At 11:37 AM -0600 8/18/05, Gayle Callis wrote: >>I think the Chemicon autofluorescence removal reagent - whatever they >>call it, is to get rid of lipofuscins - their website will have the >>information how it works. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From PMonfils <@t> Lifespan.org Fri Aug 19 10:29:24 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Mohs coverslippign air bubbles Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175B5@lsexch.lsmaster.lifespan.org> The viscosity of the mounting medium is very important. If it is too dilute, it spreads across the slide too fast, in a half second or less, often trapping air bubbles. Also, if it is too dilute, the relatively large amount of solvent, once it evaporates, will reduce the total volume of the medium enough that it no longer completely fills the space between the slide and the coverslip. Air is then drawn under the coverslip to fill the space voided by the evaporating solvent. If the medium is of proper viscosity it will spread slowly between slide and coverslip, taking perhaps 3 to 5 seconds to fill the space, with much less tendency to trap air bubbles. Also, you have more control. You can actually watch the progress of the spreading medium, and if you see an air bubble forming you can easily remove it by a little pressure on the coverslip with forceps. Also, the slide should be moist (but not dripping wet) with the clearing agent. If the slide dries too much before applying the medium and coverslip, air bubbles are more likely to form. And if too much clearing agent is present it will reduce the viscosity of the mounting medium, causing problems as mentioned above. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Dave > Johnson > Sent: Friday, August 19, 2005 7:34 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Mohs coverslippign air bubbles > > Question: > > I have mohs tech in OH that is having a lot of problems with air bubbles > when coverslipping. THey coverslip by hand and they have tried various > mouting medias, etc and still are baffled > > Any suggestions > > Dave Johnson > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From tdobersztyn <@t> chmca.org Fri Aug 19 12:27:35 2005 From: tdobersztyn <@t> chmca.org (tdobersztyn@chmca.org) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Contact lists???? Message-ID: Hello everyone. I was wondering if there was a database of HT's and/or those working in the field?? It is for educational research. My study pertains to HT Job Vacancy and compensation. (loaded topics!) If you would like to help me with this, or know of a list of HT's please advice! Thank you all so much in advance Theresa Theresa R Dobersztyn HT ASCP Electron Microscopy/Histology Labs Senior Technologist Department of Pathology Akron Children's Hospital 1 Perkins Square Akron, Ohio 44308 330-543-8279 From Jackie.O'Connor <@t> abbott.com Fri Aug 19 12:37:50 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Whole mouse liver Message-ID: Does anyone have a tried and true paraffin processing schedule for whole, intact mouse liver? I have a Shandon Citadel, no vacuum. Jackie O' From maria <@t> ski.org Fri Aug 19 12:46:23 2005 From: maria <@t> ski.org (Maria Mejia) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] histo persons at Genentech Message-ID: <43061AEF.8010209@ski.org> Hello Everyone, I was wondering if there are any histology people out there, who work at Genentech? If so, I would be most grateful if you would contact me. I have a few questions I'd like to ask. Yours Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 From hymclab <@t> hyhc.com Fri Aug 19 14:45:44 2005 From: hymclab <@t> hyhc.com (hymclab) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Xylene Substitute ? Message-ID: I have used the Shandon Xylene Substitute for approximately 13 years in all aspects that I would normally use Xylene (processing, staining, coverslipping, etc..) with the exception of the cleaning cycle on my tissue processors and soaking off old coverslips. We are very happy with it. It is odorless and not greasy feeling what-so-ever and the slides dry relatively quickly. Hope this helps, Dawn -----Original Message----- From: LeVier, Rebecca J [mailto:RJLevier@LancasterGeneral.org] Sent: Thursday, August 18, 2005 10:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Xylene Substitute ? Hello Everyone, I have a question regarding the Xylene Substitute, Xylene Substitute 2 from Shandon. What is everyone's opinion of the substitute and have you seen anything that could be wrong with using this in every step that Xylene is usually used? Thanks, Becky _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Fri Aug 19 16:36:33 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Fixative for storage of perfused rat organs Message-ID: <5784D843593D874C93E9BADCB87342AB916533@tpiserver03.Coretech-holdings.com> Tip: to preserve the extracellular space for EM of soft tissue, use the Perfusion One, and associated protocol. It was originally developed for EM. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=Sacrifice+Equipment&idsubcategory=21 Don't know about the storage question. 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 Clover Daley Sent: Thursday, August 18, 2005 4:12 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Fixative for storage of perfused rat organs I am putting a tissue bank together for research students. We are currently perfusing with 2% paraformaldehyde and 1% glutaraldehyde in phosphate buffer to collect tissues for EM studies. The liver, kidney, stomach, brain will be stored. What fixative and temperature of solution would be best. These tissues will be used for light microscopy studies. Also we will be perfusing with 4% paraformaldehyde in phosphate buffer, harvesting the same tissues. Should they be stored in the same solution and do they need to be kept in the refrigerator? Thank you for your time, Clover Daley Palmer Center for Chiropractic Research _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kotreshmattur <@t> yahoo.com Sat Aug 20 07:41:34 2005 From: kotreshmattur <@t> yahoo.com (kotresh mattur) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Re: Cryosections from formalin fixed tissues Message-ID: <20050820124134.59839.qmail@web52310.mail.yahoo.com> Hi dear all, We r trying to get cryosections from 10% NBF fixed tissues for Oil Red 'O' staining. First we kept the fixed tissues for washing under tap water for overnight and then tried cryosectioning where we didnot get the quality sections Later we tried by keeping the tissues under 20% sucrose solution under refrigeration overnight(as mentioned by Gayle callis )and then we got better quality sections compared to the above said method. still we have few querries, can anybody give us ideal procedure for the same with your practical experiences? 1. How does sucrose acts in preserving the tissues? i have read that it acts as cryoprotectant but i want to know how exactly it helps 2.If we r keeping the 10% NBF fixed tissues in 20% sucrose solution then for how many days the tissues can be kept in the same solution under ref.? If so does it affect or come in the way of Oil Red 'O' staining and evaluation? 3.After taking cryosetions if i want to preserve the same tissues whether they can be placed back to 10% NBF ? Thanks in advance Dr.K.Y.Mathur Dr.K.Y.Mathur Junior Scientist Dept.of Preclincal Safety Evaluation Discovery Research Dr.Reddy's Laboratory Ltd. Bollaram Road, Miyapur, HYDERABAD-INDIA-49. Ph No: 040-23045439-exn 424,421,422,423 Cell :09391293129 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From kotreshmattur <@t> yahoo.com Sat Aug 20 07:45:28 2005 From: kotreshmattur <@t> yahoo.com (kotresh mattur) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Re: Brain sections-rat Message-ID: <20050820124528.39673.qmail@web52305.mail.yahoo.com> Dear all, We are facing some problems in rat brain sectioning and staining. We are sectioning brain at 5-6 microns thickness and then we take sections at 37-40 centigrade after leaving for 1min. in water bath or till the sections are flattened. During staining at hydration step sections are lifting off. For this problem we tried to take the sections on histogrip coated slides which also did not help us at all. Whether there are any special delicate techniques in brain sectioning, staining? Regarding this problem can anybody help with their vast experience? Suggestions/advice from all are welcome. Thanks in advance Dr.K.Y.Mathur Dr.K.Y.Mathur Junior Scientist Dept.of Preclincal Safety Evaluation Discovery Research Dr.Reddy's Laboratory Ltd. Bollaram Road, Miyapur, HYDERABAD-INDIA-49. Ph No: 040-23045439-exn 424,421,422,423 Cell :09391293129 --------------------------------- Yahoo! Mail for Mobile Take Yahoo! Mail with you! Check email on your mobile phone. From cfavara <@t> niaid.nih.gov Sat Aug 20 09:14:22 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:28 2005 Subject: [Histonet] Re: Brain sections-rat Message-ID: I would be happy to help with suggestions but need a bit more precise information. The information provided indicates you are cutting fixed embedded tissue. Is this formalin fixed paraffin embedded, standard protocol? Your problems may be inadequate fixation/processing. I have been doing rodent tissue for years and with brain I have found that inadequate fixation leads to a myriad of problems. There have been numerous discussions of rodent brain fixation/processing/cutting over the years that you should be able to find in the archives. If you still have questions please fell free to contact me. 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: kotresh mattur [mailto:kotreshmattur@yahoo.com] Sent: Saturday, August 20, 2005 5:45 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Brain sections-rat Dear all, We are facing some problems in rat brain sectioning and staining. We are sectioning brain at 5-6 microns thickness and then we take sections at 37-40 centigrade after leaving for 1min. in water bath or till the sections are flattened. During staining at hydration step sections are lifting off. For this problem we tried to take the sections on histogrip coated slides which also did not help us at all. Whether there are any special delicate techniques in brain sectioning, staining? Regarding this problem can anybody help with their vast experience? Suggestions/advice from all are welcome. Thanks in advance Dr.K.Y.Mathur Dr.K.Y.Mathur Junior Scientist Dept.of Preclincal Safety Evaluation Discovery Research Dr.Reddy's Laboratory Ltd. Bollaram Road, Miyapur, HYDERABAD-INDIA-49. Ph No: 040-23045439-exn 424,421,422,423 Cell :09391293129 --------------------------------- Yahoo! Mail for Mobile Take Yahoo! Mail with you! Check email on your mobile phone. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Sat Aug 20 10:31:03 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Contact lists???? In-Reply-To: Message-ID: ASCP has a data base of HT/HTL who are members of ASCP, which they would be willing to sell you for a one time only use. I don't know if NSH also does this (sell lists), though of course they have a data base of NSH members. However, ASCP Board of Registry already does this survey on vacancies and salaries, and have been doing it for the last 10-12 years. It was every other year. Now they are doing it every year. Go to this web page http://www.ascp.org/bor/center/center_research.asp And you can read the results from 1998-2003. The 2004 results will probably be published late 2005 (it's always a year behind, what with gathering the stats, analyzing them, writing the article and finally getting it published.) Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of tdobersztyn@chmca.org Sent: Friday, August 19, 2005 1:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Contact lists???? Hello everyone. I was wondering if there was a database of HT's and/or those working in the field?? It is for educational research. My study pertains to HT Job Vacancy and compensation. (loaded topics!) If you would like to help me with this, or know of a list of HT's please advice! Thank you all so much in advance Theresa Theresa R Dobersztyn HT ASCP Electron Microscopy/Histology Labs Senior Technologist Department of Pathology Akron Children's Hospital 1 Perkins Square Akron, Ohio 44308 330-543-8279 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Helen.Shulver <@t> pharmagene.com Mon Aug 22 08:54:00 2005 From: Helen.Shulver <@t> pharmagene.com (Helen Shulver) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Frozen tissue array construction Message-ID: <60874D1C09E623488DF12342C80325EEC3233A@pgn-mail.pharmagene.com> Hello, I am looking to make some frozen tissue arrays in our lab. I have a tissue arrayer machine and have also read the frozen tumour paper by fejzo and Slamon. I would be extremely grateful for any help that fellow histonetters can offer in the construction of these arrays. Many thanks, Helen This message and any files transmitted with it are intended for the addressee only and may contain information that is confidential or privileged. Unauthorised use is strictly prohibited and may be unlawful. If you are not the addressee, you should not read, copy,disclose or otherwise use this message, except for the purpose of delivery to the addressee. If you received it in error please notify us immediately and then destroy it. Please note that e-mails sent to Pharmagene may be opened by an authorised delegate appointed by the addressee. Further, Pharmagene makes every effort to keep its network free from viruses. However, you do need to check this e-mail and any attachments to it for viruses as Pharmagene can take no responsibility for any computer virus that might be transferred by way of this e-mail. From PIXLEYSK <@t> UCMAIL.UC.EDU Mon Aug 22 09:26:39 2005 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] RE: Fixative for storage of perfused rat organs Message-ID: Dear Clover Daley: Depending on your antigens, I would not store in paraformaldehyde-too harsh. For storage, you can store short periods of time in phosphate buffer with sodium azide (i.e. 0.02%). You definitely need to refrigerate and add azide because otherwise they will get bacteria and/or mold. If you are going to do frozen sectioning, then I currently use and really like the following protocol for long-term storage. Supposedly this allows storage for years without loss of antigenicity. (If you are paraffin embedding, store the tissues after embedding in paraffin.) Cryoprotection and long-term storage: Preparing tissues for long-term storage prior to cryostat sectioning. * Perfuse or post-fix tissues in paraformaldehyde. * Rinse with buffer or water * Decalcify if necessary, rinse with water, then buffer. * Put tissues in Cryoprotectant (see below), Store in fridge until needed: supposedly good for years. * Before sectioning, put tissues in 30% sucrose in 0.1M PB, overnight or until tissue floats. Can keep in sucrose 1 week or so. * Remove tissue, embed in M1 embedding matrix (Shandon). * Freeze tissue in cryostat, cut cryostat sections, discard uncut tissue. Cryoprotectant: (30% ethylene glycol v/v; 30 % sucrose, 0.02% Na Azide, 0.04 M PB) Total volume: 1 liter: Add, in order indicated: 1. 200 ml dd water 2. 200 ml 0.2 M phosphate buffer (final conc. 0.04M PB) 3. 300 ml Ethylene Glycol 4. 10 ml of 2% Na Azide 5. 300 g sucrose: add slowly (~100 g at a time) with stirring 6. Stir until dissolved 7. Check volume, but should be 1 liter. 8. Refrigerate 30% Sucrose in 0.1M Phosphate Buffer: For 500 mls: 1. 250 mls 0.2 M PB 2. 150 g sucrose 3. 100 mls water 4. 5 mls 2% Na Azide (100X) 5. Stir until dissolved 6. Add water to make up to 500 mls 7. Refrigerate. Sarah Pixley University of Cincinnati College of Medicine From TJJ <@t> Stowers-Institute.org Mon Aug 22 10:30:43 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] PMMA sections sticking to plastic Message-ID: We're having some problems with our PMMA sections adhering to the plastic instead of the glass slide. After sectioning, we place the sections on a plus slide (+ charged) flooded with 50% alcohol. We stretch the section, place a strip of plastic wrap over it, and roll it flat. We will then stack 10 or so slides like this together with tongue depressors at the top and bottom, and clamp them together. We will leave them in a 60 degree oven overnight (at least). After we remove them from the oven, we are unable to peel the plastic off the slides without the sections coming off as well. The last time this happened, I proceeded with deplasticizing using xylene and MEA (recommended by the Technovit instructions) and kept the plastic wrap on the slides. I was able to remove it by eventually placing the slides in acetone. Is there an easier way? Best wishes, Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From cytologer <@t> gmail.com Mon Aug 22 11:03:20 2005 From: cytologer <@t> gmail.com (Ul Soo Choi) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] blood sample for TEM... Message-ID: Hi histonetters! I am interested and in need of a protocol for preparing blood samples for transmission electron microscopy. But I am short of appropriate protocol for fixation, and processing for it. I would like to see cat neutrophils and lymphocytes by TEM, and Would anyone there who has experiences in blood specimen for TEM, kindly give me some tips for preparing blood samples for EM? Thank you for any inputs. I would appreciate. Best regards, Ul Soo Choi DVM., PhD. Dept of clinical pathology, CVM, SNU, Seoul, Korea 82-02-880-8688 From sjchtascp <@t> yahoo.com Mon Aug 22 11:21:21 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] sucrose infiltrated muscle Message-ID: <20050822162121.23680.qmail@web90201.mail.scd.yahoo.com> Has anyone ever worked with fixed muscle infiltrated in sucrose then snap frozen. I'm looking at the option due to inconsistencies with regular snap frozen muscle? Steve --------------------------------- Start your day with Yahoo! - make it your home page From ian.montgomery <@t> bio.gla.ac.uk Mon Aug 22 11:28:05 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:25:29 2005 Subject: Fwd: [Histonet] blood sample for TEM... Message-ID: <6.2.3.4.2.20050822172223.030e61e0@udcf.gla.ac.uk> For TEM I would simply put a couple of drops of blood into the primary fixative. Agitate gently, leave for 30 minutes then spin down. Decant off the supernatant and replace with several buffer washes before the post fixative. Dehydrate and embed in epoxy resin. If the pellet started to breakdown during processing then another spin solved the problem. Ian. >Envelope-to: iom1f@udcf.gla.ac.uk >DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; s=beta; d=gmail.com; > >h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; > >b=C+bXiUq+XgmcE9ir4aw8/dBdwHo8kesoBNmC5VgG13gSwhjQQGhTv2k6y13psg87AY9Ny7XbyIyb56G9O6COjxgvToV22vy225UYEHInQpe9wJNKY9qAXbiOBnxxLGgZf6sIZghXhCkSnOYq5zdVGxRt6vUOODtKcqBa/QUTNZI= >Date: Tue, 23 Aug 2005 01:03:20 +0900 >From: Ul Soo Choi >To: histonet@lists.utsouthwestern.edu >X-Scan-Signature: 201832d891fa504721b2e650a5a52212 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >List-Id: For the exchange of information pertaining to histotechnology and > related fields >List-Unsubscribe: >, > > >List-Archive: >List-Post: >List-Help: >List-Subscribe: , > >Sender: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 05628b3a735271e1629bc35bb8abda07 >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: [Histonet] blood sample for TEM... >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.0 required=5.5 tests=none autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >X-GLA-Spam-Scan: R >X-GLA-Spam-Score: 0.0 (/) >X-GLA-Spam-Report: > >Hi histonetters! > >I am interested and in need of a protocol for preparing blood samples >for transmission electron microscopy. But I am short of appropriate >protocol for fixation, and processing for it. > >I would like to see cat neutrophils and lymphocytes by TEM, and > >Would anyone there who has experiences in blood specimen for TEM, kindly >give me some tips for preparing blood samples for EM? > >Thank you for any inputs. I would appreciate. > >Best regards, > >Ul Soo Choi >DVM., PhD. >Dept of clinical pathology, >CVM, SNU, Seoul, Korea >82-02-880-8688 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From DCadoretteHall <@t> sprg.mercy.net Mon Aug 22 11:31:33 2005 From: DCadoretteHall <@t> sprg.mercy.net (Cadorette-Hall, Diane M) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] RE: Salary Survey Message-ID: <99648DDDA90EA647B3E8A1FD4AFD339801704B11@sprgexelroy.sprg.mercy.net> Theresa, The ASCP does do a salary and vacancy survey, however, actual salaries are not reflected in those reports. For your area, Ohio in the East North Central area, the 2003 ASCP survey reported the salaries of HT from $14.43 to $18.82. Recently I was gathering this same information to take to my managers for a possible adjustment in our ranges, I received reports from HT's working in your area making from $3.19 to $5.13 more per hour than the ASCP report indicated. I also used the internet link for Bank Rate.Com Cost of Living Comparison Calculator to calculate the salaries reported by actual workers compared to the salaries reported by the ASCP. This showed that the actual salaries employers were willing to pay were higher than adjustments made for the cost of living in certain cities. I asked the NSH to help with a salary survey and was dissapointed to have them tell me that they use the ASCP reports. The recent reply to the Letters to the Editor in the August 15th issue of Advance magazine was from the president of the NSH. His reponse was, again, disappointing on several counts. What do the rest of the HistoNetters think? Diane Cadorette-Hall, HT (ASCP) Pathology Laboratory Supervisor St. Johns Regional Health Center 1235 East Cherokee Springfield, Missouri 65804 (417) 820-6492 Fax: (417) 820-7790 DCadoretteHall@sprg.mercy.net From M.Prideaux <@t> sheffield.ac.uk Mon Aug 22 11:33:52 2005 From: M.Prideaux <@t> sheffield.ac.uk (Matt Prideaux) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] PMMA sections sticking to plastic Message-ID: <1124728431.4309fe700326d@webmail.shef.ac.uk> Hi, We had a similar problem on our LR White resin sections. Have you tried leaving the slides to cool for a couple of hours after they have been removed from the oven? this seemed to solve the problem for us. Also what type of plastic wrapping do you use? we found that saran wrap seemed to work fine whereas using a thicker plastic wrap caused problems when we tried to remove it. Hope that helps Matt -- Matt Prideaux Academic Unit of Bone Biology Division of Clinical Sciences (South) D Floor (DU20) Medical School Henry Wellcome Laboratories for Medical Research University of Sheffield Beech Hill Road Sheffield S10 2RX Quoting "Johnson, Teri" : > We're having some problems with our PMMA sections adhering to the > plastic instead of the glass slide. After sectioning, we place the > sections on a plus slide (+ charged) flooded with 50% alcohol. We > stretch the section, place a strip of plastic wrap over it, and roll it > flat. We will then stack 10 or so slides like this together with tongue > depressors at the top and bottom, and clamp them together. We will > leave them in a 60 degree oven overnight (at least). After we remove > them from the oven, we are unable to peel the plastic off the slides > without the sections coming off as well. > > The last time this happened, I proceeded with deplasticizing using > xylene and MEA (recommended by the Technovit instructions) and kept the > plastic wrap on the slides. I was able to remove it by eventually > placing the slides in acetone. Is there an easier way? > > Best wishes, > > Teri Johnson > Managing Director Histology Facility > Stowers Institute for Medical Research > 1000 E. 50th St. > Kansas City, MO 64133 > > From BDUE <@t> PARTNERS.ORG Mon Aug 22 11:36:46 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] sucrose infiltrated muscle Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB50102711F@PHSXMB7.partners.org> What kind of "inconsistencies" are you experiencing? Are these clinical bxs? -brice Neuropathology Brigham & Women's Hosp. Boston "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Steven Coakley Sent: Monday, August 22, 2005 12:21 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] sucrose infiltrated muscle Has anyone ever worked with fixed muscle infiltrated in sucrose then snap frozen. I'm looking at the option due to inconsistencies with regular snap frozen muscle? Steve --------------------------------- Start your day with Yahoo! - make it your home page _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Mon Aug 22 11:38:48 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] PMMA sections sticking to plastic Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175B8@lsexch.lsmaster.lifespan.org> Some years ago, when we were first setting up for PMMA, we experimented with various kinds of plastic wraps and other polymer films. We quickly discovered that plastic wraps are not all the same. Several of them adhered to the PMMA after oven drying, just as you described. We finally settled on a product called Handi-Wrap, manufactured by Dow Chemical Co., and available at the supermarket. I have been using it ever since, and have never again encountered this problem, whether I use + slides, gelatin-coated slides, chrome-gelatin, PVA, or other adhesives. Personally though, I don't think that + slides alone, without additional adhesive, provide sufficient adhesion of the tissue to the slide in PMMA work (at least not for bone). I usually use Sta-On by Surgipath, which is a chrome-gelatin product, in an 80 degree waterbath. Otherwise my method is essentially the same as yours. Except I like pink rubber erasers between the slides and the clamp, rather than tongue depressors. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Johnson, Teri > Sent: Monday, August 22, 2005 8:30 AM > To: Histonet > Subject: [Histonet] PMMA sections sticking to plastic > > <> > We're having some problems with our PMMA sections adhering to the > plastic instead of the glass slide. After sectioning, we place the > sections on a plus slide (+ charged) flooded with 50% alcohol. We > stretch the section, place a strip of plastic wrap over it, and roll it > flat. We will then stack 10 or so slides like this together with tongue > depressors at the top and bottom, and clamp them together. We will > leave them in a 60 degree oven overnight (at least). After we remove > them from the oven, we are unable to peel the plastic off the slides > without the sections coming off as well. > > The last time this happened, I proceeded with deplasticizing using > xylene and MEA (recommended by the Technovit instructions) and kept the > plastic wrap on the slides. I was able to remove it by eventually > placing the slides in acetone. Is there an easier way? > > Best wishes, > > Teri Johnson > Managing Director Histology Facility > Stowers Institute for Medical Research > 1000 E. 50th St. > Kansas City, MO 64133 > > > From gcallis <@t> montana.edu Mon Aug 22 12:12:44 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] PMMA sections sticking to plastic In-Reply-To: <09C945920A6B654199F7A58A1D7D1FDE017175B8@lsexch.lsmaster.l ifespan.org> References: <09C945920A6B654199F7A58A1D7D1FDE017175B8@lsexch.lsmaster.lifespan.org> Message-ID: <6.0.0.22.1.20050822110621.01b16e88@gemini.msu.montana.edu> Handi wrap will not stick to superglue either - we used it as spacer between 100 um or thicker sections when supergluing these secions to a white plastic slide, then clamping or weighting down to let glue "set up". Haupts gelatin also is a common gelatin based adhesive for MMA sections but you don't need to bother with plus charge when using this, once the plus charge is coated with a protein, you have rendered it useless. You can save the expense and just use regular slides with Haupts or any other gelatin coating. I loved the pink eraser idea, gentle, softer and must conform to the surface nicely when clamped firmly. At 10:38 AM 8/22/2005, you wrote: >Some years ago, when we were first setting up for PMMA, we experimented with >various kinds of plastic wraps and other polymer films. We quickly >discovered that plastic wraps are not all the same. Several of them adhered >to the PMMA after oven drying, just as you described. We finally settled on >a product called Handi-Wrap, manufactured by Dow Chemical Co., and available >at the supermarket. I have been using it ever since, and have never again >encountered this problem, whether I use + slides, gelatin-coated slides, >chrome-gelatin, PVA, or other adhesives. > >Personally though, I don't think that + slides alone, without additional >adhesive, provide sufficient adhesion of the tissue to the slide in PMMA >work (at least not for bone). I usually use Sta-On by Surgipath, which is a >chrome-gelatin product, in an 80 degree waterbath. Otherwise my method is >essentially the same as yours. Except I like pink rubber erasers between >the slides and the clamp, rather than tongue depressors. > > > ---------- > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > > Johnson, Teri > > Sent: Monday, August 22, 2005 8:30 AM > > To: Histonet > > Subject: [Histonet] PMMA sections sticking to plastic > > > > <> > > We're having some problems with our PMMA sections adhering to the > > plastic instead of the glass slide. After sectioning, we place the > > sections on a plus slide (+ charged) flooded with 50% alcohol. We > > stretch the section, place a strip of plastic wrap over it, and roll it > > flat. We will then stack 10 or so slides like this together with tongue > > depressors at the top and bottom, and clamp them together. We will > > leave them in a 60 degree oven overnight (at least). After we remove > > them from the oven, we are unable to peel the plastic off the slides > > without the sections coming off as well. > > > > The last time this happened, I proceeded with deplasticizing using > > xylene and MEA (recommended by the Technovit instructions) and kept the > > plastic wrap on the slides. I was able to remove it by eventually > > placing the slides in acetone. Is there an easier way? > > > > Best wishes, > > > > Teri Johnson > > Managing Director Histology Facility > > Stowers Institute for Medical Research > > 1000 E. 50th St. > > Kansas City, MO 64133 > > > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From ameeker <@t> mail.jhmi.edu Mon Aug 22 12:31:44 2005 From: ameeker <@t> mail.jhmi.edu (ALAN K MEEKER) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Re: Autofluorescence reagent Message-ID: <59e6d7a64829.4309d3c0@jhmimail.jhmi.edu> From: "Andrea T. Hooper" === >>At 11:37 AM -0600 8/18/05, Gayle Callis wrote: >I think the Chemicon autofluorescence removal reagent - whatever >they call it, is to get rid of lipofuscins - their website will have >the information how it works. Has anyone used this? Have you used this yourself Gayle? It worries me that the instructions say to use it after staining and then subsequently do 70% ethanol rinses! === Andrea, I have tried the Chemicon product a couple of times. We were looking for a way to eliminate autofluorescence in probing DNA sequence using a Cy3-labelled PNA probe on various FFPE tissues. We followed the manufacturer's directions and saw a reduction in background autofluorescence to varying extents (depending on the tissue type). I'd say at best we might have lost about 1/3 of the background. Unfortunately, this was accompanied by a decrease in the fluorescent intensity of the Cy3 probe signals as well, so we really dodn't gain anything. I didn't do any controls without the reagent to see if this reduced signal was due to the reagent, of to the ethanol steps which we usually do not do in our protocol. Alan Meeker, PhD Assistant Professor of Pathology Department of Pathology Division of Genitourinary Pathology Bunting-Blaustein Cancer Research Building Room 153 1650 Orleans Street Baltimore, MD 21231-1000 ph (410) 502-7880 /(410) 614-5686 fax (410) 502-9817 From pathrm35 <@t> adelphia.net Mon Aug 22 12:32:30 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] IF antibody help Message-ID: <26499674.1124731950671.JavaMail.root@web2.mail.adelphia.net> I was wondering what company you all are using for your IF antibodies? If you could send me some catalog numbers that would be great. I'm interested in C3, fibrinogen, IgA, IgG and IgM. Thanks in advance, Ron Martin From GDawson <@t> dynacaremilwaukee.com Mon Aug 22 12:37:59 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] RE: Salary Survey Message-ID: Diane, I have found the same. I'm not sure if histotechs are not reporting their real salaries to the ASCP or what, but I have found that the payscales listed in the survey are SIGNIFICANTLY lower than what employers in Wisconsin are really paying for newly hired histotechs. The survey has actually become a tool for employers to use to justify paying histotechs less so I would strongly suggest that any techs answering this survey report their real wages. Glen Dawson -----Original Message----- From: Cadorette-Hall, Diane M [mailto:DCadoretteHall@sprg.mercy.net] Sent: Monday, August 22, 2005 10:32 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: Salary Survey Theresa, The ASCP does do a salary and vacancy survey, however, actual salaries are not reflected in those reports. For your area, Ohio in the East North Central area, the 2003 ASCP survey reported the salaries of HT from $14.43 to $18.82. Recently I was gathering this same information to take to my managers for a possible adjustment in our ranges, I received reports from HT's working in your area making from $3.19 to $5.13 more per hour than the ASCP report indicated. I also used the internet link for Bank Rate.Com Cost of Living Comparison Calculator to calculate the salaries reported by actual workers compared to the salaries reported by the ASCP. This showed that the actual salaries employers were willing to pay were higher than adjustments made for the cost of living in certain cities. I asked the NSH to help with a salary survey and was dissapointed to have them tell me that they use the ASCP reports. The recent reply to the Letters to the Editor in the August 15th issue of Advance magazine was from the president of the NSH. His reponse was, again, disappointing on several counts. What do the rest of the HistoNetters think? Diane Cadorette-Hall, HT (ASCP) Pathology Laboratory Supervisor St. Johns Regional Health Center 1235 East Cherokee Springfield, Missouri 65804 (417) 820-6492 Fax: (417) 820-7790 DCadoretteHall@sprg.mercy.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From edonato <@t> fhcrc.org Mon Aug 22 12:51:23 2005 From: edonato <@t> fhcrc.org (Donato, Elizabeth M) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] IHC position at Fred Hutchinson Cancer Research Center Message-ID: <5597E353734D6F40BB80524918D088671ECE29@harpo.fhcrc.org> Hello, We have an IHC research tech opening and I'd like to pass along the information to any interested histonet members. Here's the link to our web posting https://erecruit.fhcrc.org/servlets/iclientservlet/fhjobs/?ICType=Panel&Menu=ROLE_APPLICANT&Market=GBL&PanelGroupName=HR_JOB_POST_APP . Please let me know if you are interested or if you have any questions about the position. Liz Donato Fred Hutchinson Cancer Research Center PO Box 19024, Mail Stop C1-015 1100 Fairview Ave N. Seattle WA 98109-1024 edonato@fhcrc.org 206.667.4501 ph 206.667.5815 fax From John.Thweatt <@t> med.va.gov Mon Aug 22 12:57:04 2005 From: John.Thweatt <@t> med.va.gov (Thweatt, John T) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] LAP Testing Message-ID: <7A00E2723D55D311AC240000F831866802606FA2@VHAAMAEXC1.amarillo.med.va.gov> Dear Fellow Histonetters, Just curious, but does anyone run or do the LAP stain better known as the Leukocyte Alkaline Phosphatase test? Is this something that Histology does routinely in Histology or is this a function shared or done by other departments, or did it get delegated to Histology like it did in our lab for one reason or another? Is our lab unique that we would do this test?, or is this a common practice? It seems more hematological than histological, yet I can see the staining application and how it would fall into histology. Just wondering? Thanks in advance. See some of you all in Florida. Sincerely, Tom Thweatt, HT (ASCP) Amarillo VA Health Care System Pathology and Laboratory Medicine Service Anatomical Pathology 6010 Amarillo Boulvard West Amarillo, TX 79106 806-355-9703 x 7071 fax 806-354-7865 From BDUE <@t> PARTNERS.ORG Mon Aug 22 12:59:35 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] sucrose infiltrated muscle Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB501027120@PHSXMB7.partners.org> Steve, You didn't mention what the problems have been, nor which "snap" freeze protocol you've been using. But even whole adult rat quads should not be too big to snap freeze. Nor whole limbs. I've frozen entire rat pups without artifact. Human bxs up to 2.0x2.0x3.0cm without freeze atrifacts (those were autopsy steaks). Are you using LN2 chilled isopentane? If so, try letting the isopentane solidify or almost solidify. Then remove the isopentane in the metal beaker and break it up as it thaws. You only get about 2min to work depending on how much isopentane you are using. I get the best (quickest) freezes when the isopentane is a chunky slush. The solid bits act as a heatsink so the liquid can stay at freezing point longer. When you dunk the tissue be careful about bumping the chunks during the first 0.5-1.5sec, then agitate agitate agitate. If you don't agitate, the isopentane immediately surrounding the tissue will have warmed up and do you no good. Near freezing pt, the liquid is too thick to convect well. So agitate. Time to freeze all the way to center is never certain, so I always spend 20-30sec even for small bxs. Once all the isopentane chunks are gone is one way of timing the freeze. This can cause the tissue to crack, but you can either learn to cut cracked tissue or repair the cracks with cold OCT (refrigerator temp or lower). You can't repair freeze artifacts once they happen, so choose your trade-off. Are you sure your freeze artifacts (if that is what you are getting) are happening at the time of snap freeze? IF the tissue ever thaws partially then ice crystals will grow as it re-freezes. Check all your handling to eliminate any possibility of thaw. Especially: how do you mount the tissue for sectioning? Do you use warm room temp OCT? Any source of heat can cause problems, even long after the snap freeze. Good luck, -brice "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: Steven Coakley [mailto:sjchtascp@yahoo.com] Sent: Monday, August 22, 2005 1:30 PM To: Due, Brice Subject: RE: [Histonet] sucrose infiltrated muscle Rat/mouse leg muscle for research. Often a "tad" to big for good snap frozens. "Due, Brice" wrote: What kind of "inconsistencies" are you experiencing? Are these clinical bxs? -brice Neuropathology Brigham & Women's Hosp. Boston "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Steven Coakley Sent: Monday, August 22, 2005 12:21 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] sucrose infiltrated muscle Has anyone ever worked with fixed muscle infiltrated in sucrose then snap frozen. I'm looking at the option due to inconsistencies with regular snap frozen muscle? Steve --------------------------------- Start your day with Yahoo! - make it your home page _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____ Start your day with Yahoo! - make it your home page From PMonfils <@t> Lifespan.org Mon Aug 22 13:02:22 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Re: Cryosections from formalin fixed tissues Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175B9@lsexch.lsmaster.lifespan.org> Sucrose doesn't "preserve" the tissue in the same sense that fixatives do. Rather it protects the structural integrity of the tissue which would otherwise be disrupted by the formation of ice crystals. A concentrated sucrose solution freezes as a microscopically homogeneous matrix, without the formation of large crystals as seen during the freezing of most aqueous solutions. This helps in two ways. First, ice crystals as they form perforate cell membranes and other delicate structures, and also tend to "push aside" the surrounding tissue, leaving spaces once the ice eventually melts. Secondly, ice does not slice smoothly, so the more ice is present in a tissue, the more difficult it will be to produce a smooth slice of that tissue. The sucrose matrix slices much more smoothly than ice does. Tissue that has been in formalin or another aqueous fixative (or which has been washed in water or buffer) naturally contains a large amount of water, and therefore freezes with a high degree of ice formation. I have occasionally kept tissues in sucrose solution for several weeks without adverse effects, though I don't do so routinely. About the only possible problem that might occur would be the growth of microorganisms in the solution; but even that is unlikely due to the high osmotic pressure of such a solution (we don't have to refrigerate corn syrup or honey, for example). Though I have not tested Oil Red O staining after long sucrose exposure, it seems unlikely that there would be an adverse effect since fat is immiscible with aqueous solutions, and I can't think of any actual reaction that might occur between the lipids and the sucrose. Sections can be returned to formalin after cryosectioning with sucrose, and if the tissue was well infiltrated with the sucrose solution and properly frozen, there is typically minimal if any detectable freeze artifact. We did this most recently with some whole rat brains which had been sucrose-infiltrated and frozen, which we subsequently needed to embed in paraffin. If the tissue is frozen in a tissue freezing medium, I just place the whole frozen block into the formalin, and allow it to thaw there at room temparature. I tried thawing them in the refrigerator, but found that the tissue embedding medium was reluctant to dissolve into the formalin at that temperature. I leave them in formalin overnight, both to allow the embedding medium to dissolve, and to allow the sucrose solution to diffuse out of the tissue. Then I transfer them to fresh formalin. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > kotresh mattur > Sent: Saturday, August 20, 2005 5:41 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Cryosections from formalin fixed tissues > > Hi dear all, > We r trying to get cryosections from 10% NBF fixed tissues for Oil Red > 'O' staining. > > First we kept the fixed tissues for washing under tap water for > overnight and then tried cryosectioning where we didnot get the quality > sections > > Later we tried by keeping the tissues under 20% sucrose solution under > refrigeration overnight(as mentioned by Gayle callis )and then we got > better quality sections compared to the above said method. > > still we have few querries, can anybody give us ideal procedure for the > same with your practical experiences? > > 1. How does sucrose acts in preserving the tissues? i have read that it > acts as cryoprotectant but i want to know how exactly it helps > > 2.If we r keeping the 10% NBF fixed tissues in 20% sucrose solution then > for how many days the tissues can be kept in the same solution under ref.? > If so does it affect or come in the way of Oil Red 'O' staining and > evaluation? > > 3.After taking cryosetions if i want to preserve the same tissues whether > they can be placed back to 10% NBF ? > > Thanks in advance > > Dr.K.Y.Mathur > > > > Dr.K.Y.Mathur > Junior Scientist > Dept.of Preclincal Safety Evaluation > Discovery Research > Dr.Reddy's Laboratory Ltd. > Bollaram Road, Miyapur, > HYDERABAD-INDIA-49. > > Ph No: > 040-23045439-exn 424,421,422,423 > Cell :09391293129 > > __________________________________________________ > 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 Charles.Embrey <@t> carle.com Mon Aug 22 13:36:29 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] RE: Salary Survey Message-ID: One thing I was told is that ASCP only surveys technicians that are ACTIVE members of ASCP and not all registered Histotechs. If you don't pay the annual dues you don't get surveyed. This could explain the false numbers by eliminating a large portion of the histology work force. Here, only one out of my seven histotechs is an active member. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dawson, Glen Sent: Monday, August 22, 2005 12:38 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] RE: Salary Survey Diane, I have found the same. I'm not sure if histotechs are not reporting their real salaries to the ASCP or what, but I have found that the payscales listed in the survey are SIGNIFICANTLY lower than what employers in Wisconsin are really paying for newly hired histotechs. The survey has actually become a tool for employers to use to justify paying histotechs less so I would strongly suggest that any techs answering this survey report their real wages. Glen Dawson -----Original Message----- From: Cadorette-Hall, Diane M [mailto:DCadoretteHall@sprg.mercy.net] Sent: Monday, August 22, 2005 10:32 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: Salary Survey Theresa, The ASCP does do a salary and vacancy survey, however, actual salaries are not reflected in those reports. For your area, Ohio in the East North Central area, the 2003 ASCP survey reported the salaries of HT from $14.43 to $18.82. Recently I was gathering this same information to take to my managers for a possible adjustment in our ranges, I received reports from HT's working in your area making from $3.19 to $5.13 more per hour than the ASCP report indicated. I also used the internet link for Bank Rate.Com Cost of Living Comparison Calculator to calculate the salaries reported by actual workers compared to the salaries reported by the ASCP. This showed that the actual salaries employers were willing to pay were higher than adjustments made for the cost of living in certain cities. I asked the NSH to help with a salary survey and was dissapointed to have them tell me that they use the ASCP reports. The recent reply to the Letters to the Editor in the August 15th issue of Advance magazine was from the president of the NSH. His reponse was, again, disappointing on several counts. What do the rest of the HistoNetters think? Diane Cadorette-Hall, HT (ASCP) Pathology Laboratory Supervisor St. Johns Regional Health Center 1235 East Cherokee Springfield, Missouri 65804 (417) 820-6492 Fax: (417) 820-7790 DCadoretteHall@sprg.mercy.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Mon Aug 22 14:10:37 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] RE: Salary Survey In-Reply-To: Message-ID: <200508221910.j7MJAUnQ006150@chip.viawest.net> I think that makes a good argument for keeping up the ASCP membership, since ASCP is our certifing agency (not NSH) I would think that it would pay to stay connected with ASCP so that you might could have a say in such things as "salary surveys". The most critical of an organization are usually the ones least involved in that organization. It seems that it is a lot easier to criticise than to take command and make a difference. Best regards, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, August 22, 2005 11:36 AM To: Dawson, Glen; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Salary Survey One thing I was told is that ASCP only surveys technicians that are ACTIVE members of ASCP and not all registered Histotechs. If you don't pay the annual dues you don't get surveyed. This could explain the false numbers by eliminating a large portion of the histology work force. Here, only one out of my seven histotechs is an active member. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dawson, Glen Sent: Monday, August 22, 2005 12:38 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] RE: Salary Survey Diane, I have found the same. I'm not sure if histotechs are not reporting their real salaries to the ASCP or what, but I have found that the payscales listed in the survey are SIGNIFICANTLY lower than what employers in Wisconsin are really paying for newly hired histotechs. The survey has actually become a tool for employers to use to justify paying histotechs less so I would strongly suggest that any techs answering this survey report their real wages. Glen Dawson -----Original Message----- From: Cadorette-Hall, Diane M [mailto:DCadoretteHall@sprg.mercy.net] Sent: Monday, August 22, 2005 10:32 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: Salary Survey Theresa, The ASCP does do a salary and vacancy survey, however, actual salaries are not reflected in those reports. For your area, Ohio in the East North Central area, the 2003 ASCP survey reported the salaries of HT from $14.43 to $18.82. Recently I was gathering this same information to take to my managers for a possible adjustment in our ranges, I received reports from HT's working in your area making from $3.19 to $5.13 more per hour than the ASCP report indicated. I also used the internet link for Bank Rate.Com Cost of Living Comparison Calculator to calculate the salaries reported by actual workers compared to the salaries reported by the ASCP. This showed that the actual salaries employers were willing to pay were higher than adjustments made for the cost of living in certain cities. I asked the NSH to help with a salary survey and was dissapointed to have them tell me that they use the ASCP reports. The recent reply to the Letters to the Editor in the August 15th issue of Advance magazine was from the president of the NSH. His reponse was, again, disappointing on several counts. What do the rest of the HistoNetters think? Diane Cadorette-Hall, HT (ASCP) Pathology Laboratory Supervisor St. Johns Regional Health Center 1235 East Cherokee Springfield, Missouri 65804 (417) 820-6492 Fax: (417) 820-7790 DCadoretteHall@sprg.mercy.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MSafron <@t> wilresearch.com Mon Aug 22 14:32:54 2005 From: MSafron <@t> wilresearch.com (Mike Safron) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Headache X pregnancy Message-ID: If you are pregnant, or employ a tech who is, please take a look at the results of this toxicology study. I think that xylene use by those pregnant is a major issue in histology. Pregnancy Outcome Following Gestational Exposure to Organic Solvents: A Prospective Controlled Study JAMA, Mar 1999; 281: 1106 - 1109. Michael Safron A.S.,HT(ASCP) Manager, Histology WIL Research Laboratories LLC From tpmorken <@t> labvision.com Mon Aug 22 14:39:33 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:29 2005 Subject: wage survey source...[Histonet] RE: Salary Survey Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D04F@usca0082k08.labvision.apogent.com> The ASCP 2003 Wage survey article ( http://www.ascp.org/bor/center/center_research.asp ) says a "A representative sample of clinical laboratories by facility type was drawn from the Centers for Medicare and Medicaid database of all legally operating licensed clinical laboratories in the United States. Of the 33,674 laboratories in this database, 9,738 facilities were selected to participate in the survey. A total of 1,682 laboratories responded to the survey yielding a response rate of 17%. This number of respondents provides an overall sampling margin of error of +/- 2.4% at the 95% confidence level for total sample statistics at the national level." However, it goes on to say not all specialities are at all lab locations so response rate and margin of error varies between specialties. They don't say in the article, but I seem to remember that the survey is sent to the lab management, not individual technologists. The authors don't provide the quesitonaire so we don't know what was asked, whether individual salaries or averages of a given group. Anyway, there are always outliers not caugt in in any survey. An anecdotal knowledge of a given lab that pays higher than a survey is a certainty. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, August 22, 2005 11:36 AM To: Dawson, Glen; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Salary Survey One thing I was told is that ASCP only surveys technicians that are ACTIVE members of ASCP and not all registered Histotechs. If you don't pay the annual dues you don't get surveyed. This could explain the false numbers by eliminating a large portion of the histology work force. Here, only one out of my seven histotechs is an active member. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dawson, Glen Sent: Monday, August 22, 2005 12:38 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] RE: Salary Survey Diane, I have found the same. I'm not sure if histotechs are not reporting their real salaries to the ASCP or what, but I have found that the payscales listed in the survey are SIGNIFICANTLY lower than what employers in Wisconsin are really paying for newly hired histotechs. The survey has actually become a tool for employers to use to justify paying histotechs less so I would strongly suggest that any techs answering this survey report their real wages. Glen Dawson -----Original Message----- From: Cadorette-Hall, Diane M [mailto:DCadoretteHall@sprg.mercy.net] Sent: Monday, August 22, 2005 10:32 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: Salary Survey Theresa, The ASCP does do a salary and vacancy survey, however, actual salaries are not reflected in those reports. For your area, Ohio in the East North Central area, the 2003 ASCP survey reported the salaries of HT from $14.43 to $18.82. Recently I was gathering this same information to take to my managers for a possible adjustment in our ranges, I received reports from HT's working in your area making from $3.19 to $5.13 more per hour than the ASCP report indicated. I also used the internet link for Bank Rate.Com Cost of Living Comparison Calculator to calculate the salaries reported by actual workers compared to the salaries reported by the ASCP. This showed that the actual salaries employers were willing to pay were higher than adjustments made for the cost of living in certain cities. I asked the NSH to help with a salary survey and was dissapointed to have them tell me that they use the ASCP reports. The recent reply to the Letters to the Editor in the August 15th issue of Advance magazine was from the president of the NSH. His reponse was, again, disappointing on several counts. What do the rest of the HistoNetters think? Diane Cadorette-Hall, HT (ASCP) Pathology Laboratory Supervisor St. Johns Regional Health Center 1235 East Cherokee Springfield, Missouri 65804 (417) 820-6492 Fax: (417) 820-7790 DCadoretteHall@sprg.mercy.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From info <@t> instrumedics.com Mon Aug 22 14:40:16 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Frozen tissue array construction References: <60874D1C09E623488DF12342C80325EEC3233A@pgn-mail.pharmagene.com> Message-ID: <02dd01c5a751$59b39ca0$6401a8c0@INSTRUMEDICS22> Helen, As you may have read in the Fejzo/Slamon paper they were not successful until they used the CryoJane Tape-transfer system. Without snap-freezing the tissue and the tape-transfer process the cryoarrays were not acceptable. Once they used the CryoJane the results could be published. Please visit www.instrumedics.com for the details. Bernice ----- Original Message ----- From: "Helen Shulver" To: Sent: Monday, August 22, 2005 9:54 AM Subject: [Histonet] Frozen tissue array construction Hello, I am looking to make some frozen tissue arrays in our lab. I have a tissue arrayer machine and have also read the frozen tumour paper by fejzo and Slamon. I would be extremely grateful for any help that fellow histonetters can offer in the construction of these arrays. Many thanks, Helen This message and any files transmitted with it are intended for the addressee only and may contain information that is confidential or privileged. Unauthorised use is strictly prohibited and may be unlawful. If you are not the addressee, you should not read, copy,disclose or otherwise use this message, except for the purpose of delivery to the addressee. If you received it in error please notify us immediately and then destroy it. Please note that e-mails sent to Pharmagene may be opened by an authorised delegate appointed by the addressee. Further, Pharmagene makes every effort to keep its network free from viruses. However, you do need to check this e-mail and any attachments to it for viruses as Pharmagene can take no responsibility for any computer virus that might be transferred by way of this e-mail. From tpmorken <@t> labvision.com Mon Aug 22 14:50:04 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:29 2005 Subject: Clarification...wage survey source...[Histonet] RE: Salary Su rvey Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D050@usca0082k08.labvision.apogent.com> OK, I went and looked at the 2002 ASCP survey, part 1 (same link as below). In that one they detail more clearly who they sent the survey to: "The sample represented all laboratory managers who were in the ASCP membership database." However, this 2002 article says nothing about using the "...Centers for Medicare and Medicaid database of all legally operating licensed clinical laboratories in the United States. " So maybe that is a new change. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Monday, August 22, 2005 12:40 PM To: histonet@lists.utsouthwestern.edu Subject: RE: wage survey source...[Histonet] RE: Salary Survey The ASCP 2003 Wage survey article ( http://www.ascp.org/bor/center/center_research.asp ) says a "A representative sample of clinical laboratories by facility type was drawn from the Centers for Medicare and Medicaid database of all legally operating licensed clinical laboratories in the United States. Of the 33,674 laboratories in this database, 9,738 facilities were selected to participate in the survey. A total of 1,682 laboratories responded to the survey yielding a response rate of 17%. This number of respondents provides an overall sampling margin of error of +/- 2.4% at the 95% confidence level for total sample statistics at the national level." However, it goes on to say not all specialities are at all lab locations so response rate and margin of error varies between specialties. They don't say in the article, but I seem to remember that the survey is sent to the lab management, not individual technologists. The authors don't provide the quesitonaire so we don't know what was asked, whether individual salaries or averages of a given group. Anyway, there are always outliers not caugt in in any survey. An anecdotal knowledge of a given lab that pays higher than a survey is a certainty. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, August 22, 2005 11:36 AM To: Dawson, Glen; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Salary Survey One thing I was told is that ASCP only surveys technicians that are ACTIVE members of ASCP and not all registered Histotechs. If you don't pay the annual dues you don't get surveyed. This could explain the false numbers by eliminating a large portion of the histology work force. Here, only one out of my seven histotechs is an active member. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dawson, Glen Sent: Monday, August 22, 2005 12:38 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] RE: Salary Survey Diane, I have found the same. I'm not sure if histotechs are not reporting their real salaries to the ASCP or what, but I have found that the payscales listed in the survey are SIGNIFICANTLY lower than what employers in Wisconsin are really paying for newly hired histotechs. The survey has actually become a tool for employers to use to justify paying histotechs less so I would strongly suggest that any techs answering this survey report their real wages. Glen Dawson -----Original Message----- From: Cadorette-Hall, Diane M [mailto:DCadoretteHall@sprg.mercy.net] Sent: Monday, August 22, 2005 10:32 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: Salary Survey Theresa, The ASCP does do a salary and vacancy survey, however, actual salaries are not reflected in those reports. For your area, Ohio in the East North Central area, the 2003 ASCP survey reported the salaries of HT from $14.43 to $18.82. Recently I was gathering this same information to take to my managers for a possible adjustment in our ranges, I received reports from HT's working in your area making from $3.19 to $5.13 more per hour than the ASCP report indicated. I also used the internet link for Bank Rate.Com Cost of Living Comparison Calculator to calculate the salaries reported by actual workers compared to the salaries reported by the ASCP. This showed that the actual salaries employers were willing to pay were higher than adjustments made for the cost of living in certain cities. I asked the NSH to help with a salary survey and was dissapointed to have them tell me that they use the ASCP reports. The recent reply to the Letters to the Editor in the August 15th issue of Advance magazine was from the president of the NSH. His reponse was, again, disappointing on several counts. What do the rest of the HistoNetters think? Diane Cadorette-Hall, HT (ASCP) Pathology Laboratory Supervisor St. Johns Regional Health Center 1235 East Cherokee Springfield, Missouri 65804 (417) 820-6492 Fax: (417) 820-7790 DCadoretteHall@sprg.mercy.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@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 koellinr <@t> amgen.com Mon Aug 22 15:28:16 2005 From: koellinr <@t> amgen.com (Koelling, Ray) Date: Fri Sep 16 15:25:29 2005 Subject: Clarification...wage survey source...[Histonet] RE: Salary Su rvey Message-ID: <16834C6DFFA6004C88DE4507FB8AE544018CC89B@wa-mb4-sea.amgen.com> In my opinion, all the responses have been valid and rational. I've been in ASCP actively and yes it is anecdotal but I've never received a survey as an individual or a manager. Outliers in this survey must surely be more than a few. That database of salaries simply does not fully reflect reality and the real world....in my opinion. And by way, way more than statistical error from what I've seen and experienced and know. Perhaps it is time for a different database to be generated and in a completely different way by different people since the data, while it might be statistically and technically correct, is prejudicial and logically incorrect. That's not a solution-just an observation. Ray Raymond Koelling Research Scientist, Pathology Amgen Corp. Seattle -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Monday, August 22, 2005 12:50 PM To: histonet@lists.utsouthwestern.edu Subject: RE: Clarification...wage survey source...[Histonet] RE: Salary Su rvey OK, I went and looked at the 2002 ASCP survey, part 1 (same link as below). In that one they detail more clearly who they sent the survey to: "The sample represented all laboratory managers who were in the ASCP membership database." However, this 2002 article says nothing about using the "...Centers for Medicare and Medicaid database of all legally operating licensed clinical laboratories in the United States. " So maybe that is a new change. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Morken, Tim - Labvision Sent: Monday, August 22, 2005 12:40 PM To: histonet@lists.utsouthwestern.edu Subject: RE: wage survey source...[Histonet] RE: Salary Survey The ASCP 2003 Wage survey article ( http://www.ascp.org/bor/center/center_research.asp ) says a "A representative sample of clinical laboratories by facility type was drawn from the Centers for Medicare and Medicaid database of all legally operating licensed clinical laboratories in the United States. Of the 33,674 laboratories in this database, 9,738 facilities were selected to participate in the survey. A total of 1,682 laboratories responded to the survey yielding a response rate of 17%. This number of respondents provides an overall sampling margin of error of +/- 2.4% at the 95% confidence level for total sample statistics at the national level." However, it goes on to say not all specialities are at all lab locations so response rate and margin of error varies between specialties. They don't say in the article, but I seem to remember that the survey is sent to the lab management, not individual technologists. The authors don't provide the quesitonaire so we don't know what was asked, whether individual salaries or averages of a given group. Anyway, there are always outliers not caugt in in any survey. An anecdotal knowledge of a given lab that pays higher than a survey is a certainty. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, August 22, 2005 11:36 AM To: Dawson, Glen; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Salary Survey One thing I was told is that ASCP only surveys technicians that are ACTIVE members of ASCP and not all registered Histotechs. If you don't pay the annual dues you don't get surveyed. This could explain the false numbers by eliminating a large portion of the histology work force. Here, only one out of my seven histotechs is an active member. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dawson, Glen Sent: Monday, August 22, 2005 12:38 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] RE: Salary Survey Diane, I have found the same. I'm not sure if histotechs are not reporting their real salaries to the ASCP or what, but I have found that the payscales listed in the survey are SIGNIFICANTLY lower than what employers in Wisconsin are really paying for newly hired histotechs. The survey has actually become a tool for employers to use to justify paying histotechs less so I would strongly suggest that any techs answering this survey report their real wages. Glen Dawson -----Original Message----- From: Cadorette-Hall, Diane M [mailto:DCadoretteHall@sprg.mercy.net] Sent: Monday, August 22, 2005 10:32 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: Salary Survey Theresa, The ASCP does do a salary and vacancy survey, however, actual salaries are not reflected in those reports. For your area, Ohio in the East North Central area, the 2003 ASCP survey reported the salaries of HT from $14.43 to $18.82. Recently I was gathering this same information to take to my managers for a possible adjustment in our ranges, I received reports from HT's working in your area making from $3.19 to $5.13 more per hour than the ASCP report indicated. I also used the internet link for Bank Rate.Com Cost of Living Comparison Calculator to calculate the salaries reported by actual workers compared to the salaries reported by the ASCP. This showed that the actual salaries employers were willing to pay were higher than adjustments made for the cost of living in certain cities. I asked the NSH to help with a salary survey and was dissapointed to have them tell me that they use the ASCP reports. The recent reply to the Letters to the Editor in the August 15th issue of Advance magazine was from the president of the NSH. His reponse was, again, disappointing on several counts. What do the rest of the HistoNetters think? Diane Cadorette-Hall, HT (ASCP) Pathology Laboratory Supervisor St. Johns Regional Health Center 1235 East Cherokee Springfield, Missouri 65804 (417) 820-6492 Fax: (417) 820-7790 DCadoretteHall@sprg.mercy.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@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 h.croes <@t> ru.nl Mon Aug 22 15:45:27 2005 From: h.croes <@t> ru.nl (Home) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] blood sample for TEM... References: Message-ID: <004b01c5a75a$6db94200$6502a8c0@medion> Hello Ul Soo Choi, What you can do is, spin down your heparinized blood cells (unfixed). What you get is a buffycoat (white cells) on top of your erythrocytes. Gently remove the serum and then ad gently your fixative (2% glutaraldehyde/ buffer) without disturbing the buffycoat layer, let it stand for at least 3 hours on ice and then you can remove the buffycoat with your white cells as a compact layer. You can cut it in pieces and proces it like you would normally do with tissue samples. Huib ----- Original Message ----- From: "Ul Soo Choi" To: Sent: Monday, August 22, 2005 6:03 PM Subject: [Histonet] blood sample for TEM... Hi histonetters! I am interested and in need of a protocol for preparing blood samples for transmission electron microscopy. But I am short of appropriate protocol for fixation, and processing for it. I would like to see cat neutrophils and lymphocytes by TEM, and Would anyone there who has experiences in blood specimen for TEM, kindly give me some tips for preparing blood samples for EM? Thank you for any inputs. I would appreciate. Best regards, Ul Soo Choi DVM., PhD. Dept of clinical pathology, CVM, SNU, Seoul, Korea 82-02-880-8688 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From debbiekeith <@t> cox.net Mon Aug 22 16:30:02 2005 From: debbiekeith <@t> cox.net (Debbie Keith) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] update about Alysa Anne Barbarisi Message-ID: <5.2.0.9.0.20050822125009.05248b50@pop.central.cox.net> Dear Histonetters! this is an update about Lisa Barbarisi's Daughter. she was diagnosed a few months ago with stage IV Neuroblastoma. today she goes in for the 7th session of Chemo. on the 30th of this month she turns 1 year old! she has been fighting this horrible disease for half of her life, now. We have put together a benefit in downtown Phoenix at Alice Cooper'stown. Please take a moment to check out Alysa Anne's website... www.alysa-anne.com here is the page specific to the benifit http://www.alysa-anne.com/Benefit%20at%20cooper'stown.htm we are looking for ideas for the benefit. we're looking for anyone that can print t-shirts (about 100) and flyers for free. we've got news coverage (radio and tv) and we're working on local publications. if anyone has any ideas.. thoughts... donations... we would so happy! thank you! debbie keith From cytologer <@t> gmail.com Mon Aug 22 20:45:39 2005 From: cytologer <@t> gmail.com (Ul Soo Choi) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] blood sample for TEM... In-Reply-To: <004b01c5a75a$6db94200$6502a8c0@medion> References: <004b01c5a75a$6db94200$6502a8c0@medion> Message-ID: hi! dear histonetters, thank a lot for your kindness in all your quick, and detailed replies. Everytime I rely on you, I am greatly benefited. Thanks again. Ul Soo On 8/23/05, Home wrote: > Hello Ul Soo Choi, > > What you can do is, spin down your heparinized blood cells (unfixed). > What you get is a buffycoat (white cells) on top of your erythrocytes. > Gently remove the serum and then ad gently your fixative (2% glutaraldehyde/ > buffer) without disturbing the buffycoat layer, let it stand for at least 3 > hours on ice and then you can remove the buffycoat with your white cells as > a compact layer. You can cut it in pieces and proces it like you would > normally do with tissue samples. > > Huib > > ----- Original Message ----- > From: "Ul Soo Choi" > To: > Sent: Monday, August 22, 2005 6:03 PM > Subject: [Histonet] blood sample for TEM... > > > Hi histonetters! > > I am interested and in need of a protocol for preparing blood samples > for transmission electron microscopy. But I am short of appropriate > protocol for fixation, and processing for it. > > I would like to see cat neutrophils and lymphocytes by TEM, and > > Would anyone there who has experiences in blood specimen for TEM, kindly > give me some tips for preparing blood samples for EM? > > Thank you for any inputs. I would appreciate. > > Best regards, > > Ul Soo Choi > DVM., PhD. > Dept of clinical pathology, > CVM, SNU, Seoul, Korea > 82-02-880-8688 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From essssy <@t> gmail.com Mon Aug 22 21:44:37 2005 From: essssy <@t> gmail.com (DrEssy) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] automated tissue processor Message-ID: <85b73765050822194439be81f3@mail.gmail.com> Hello, Does anyone know if there is an off-the-shelf automated tissue prcessor that can be used for methacrylate embedding for 2-3 cm bone samples? Specifically, I am looking for something that has no heating capable of operating at ambient temps with the ability to draw pulsatile vacuum for better polymer penetration. Thanks in advance histonet users! -Essy From petepath <@t> yahoo.com Tue Aug 23 06:54:06 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] update about Alysa Anne Barbarisi Message-ID: <20050823115406.70753.qmail@web30414.mail.mud.yahoo.com> For anyone interested in helping Alysa Ann, I am offering her a 15% commission on sales of my Precision Cryoembedding System for anyone mentioning her name. You can demo this apparatus without obligation. If you decide to buy it, 15% of the purchase price will go to Lisa and Alysa Ann. If you want to see my system in action visit booth 835 at NSH. I challenge other vendors to get involved. Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From sobrien <@t> bthosp.com Tue Aug 23 08:09:30 2005 From: sobrien <@t> bthosp.com (O'Brien, Sue) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Autopsy Worksheet Message-ID: <4D95C24EC0E4C84787A6919F1165B25E24B1B6@btmhems01.BTHOSP.INT> Does anyone have an autopsy worksheet that they are willing to share with me? (I am looking for one that would be a fill-in the blanks type for organs, weights etc.). Anything you would have as a start point would be appreciated! Thank-you, Sue O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital E-mail: sobrien@bthosp.com From tp2 <@t> medicine.wisc.edu Tue Aug 23 09:04:27 2005 From: tp2 <@t> medicine.wisc.edu (Thomas Pier) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] CD4, CD8 antibody question Message-ID: Hello, I would like to know what CD4 and CD8 antibodies (clones and manufacturers) all of you histonetters are using with good results in FFPE tissues. Protocols would be great too if you're willing to share. Yes, I am still having difficulties with CD4 for whatever reason. I'm probably going to go back to the drawingboard here and start from the ground up again. I'd really appreciate any help that anyone could send my way. Have a great week. Tom Pier From contact <@t> excaliburpathology.com Tue Aug 23 09:16:21 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] automated tissue processor for plastics Message-ID: <20050823141622.52842.qmail@web50308.mail.yahoo.com> EMS has a processor that will take specimens all the way to 100% resin. Here is the link: http://www.emsdiasum.com/microscopy/products/equipment/tissue_processor.aspx?mm=19 Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From gcallis <@t> montana.edu Tue Aug 23 09:22:34 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Methacrylate with Re: automated tissue processor In-Reply-To: <85b73765050822194439be81f3@mail.gmail.com> References: <85b73765050822194439be81f3@mail.gmail.com> Message-ID: <6.0.0.22.1.20050823080342.01af6908@gemini.msu.montana.edu> By methacrylate, you mean methyl methacrylate? If so, the answer is No (so far no one has developed a processor for MMA protocol) but you can use an automated processor through all the dehydrationa and clearing steps as long as you do NOT use monomer as a clearing agent. We used our VIP to dehydrate through alcohols and clearing agents but programmed out all the paraffin steps. After the last clearing agent, undecalcified bone samples are removed for hand processing through infiltration mixtures. Since infiltration was done in a refrigerator at 4C to prevent unexpected polymerization of partially infiltrated samples, we simply used a Wheaton O ring vacuum dessicator with inhouse water vacuum (very handy!) or vacuum pump inside a hood with each change of infiltration mixture (we had three) and vacuum and watch bubbles come off generally for about 4 hours, then release and put the samples in a refrigerator in tightly capped containers, then remove and bring to room temperature AFTER desired infiltration time, and change mixture, repeat this process. We processed 1 cm thick bone slabs with this method, taking extremely long times for dehydration/clearing - and the VIP never missed a beat other than having to top off alcohols and clearing agents due to evaporation. Monomer was never put on this instrument. No company has developed an automated processor for methyl methacrylate but more importantly, you do NOT want to breathe the fumes as methyl methacrylate monomer is a toxic substance very damaging to Central Nervous System. If you smell it, you are exposed and one should always do any handling/infiltrations/vacuuming inside a good fume hood. Protect yourself and others from fumes - the act of changing the monomer out of a processor, disposal, etc would be a messy, smelly process. Besides you do not want some of the infiltration mixtures on the processor, if they polymerize, it would be a disaster. I will be happy to send you a very nice MMA protocol per Sterchi publication where the infiltration mixtures are NOT thick, gooey consistency, and you get excellent results for infiltration, embedding and polymerization. At 08:44 PM 8/22/2005, you wrote: >Hello, > >Does anyone know if there is an off-the-shelf automated tissue prcessor that >can be used for methacrylate embedding for 2-3 cm bone samples? >Specifically, I am looking for something that has no heating capable of >operating at ambient temps with the ability to draw pulsatile vacuum for >better polymer penetration. > >Thanks in advance histonet users! > >-Essy >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From tp2 <@t> medicine.wisc.edu Tue Aug 23 09:32:41 2005 From: tp2 <@t> medicine.wisc.edu (Thomas Pier) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] CD4, CD8 antibody question REVISED Message-ID: Hello, I would like to know what CD4 and CD8 antibodies (clones and manufacturers) all of you histonetters are using on human tissues with good results in FFPE tissues. Protocols would be great too if you're willing to share. Yes, I am still having difficulties with CD4 for whatever reason. I'm probably going to go back to the drawingboard here and start from the ground up again. I'd really appreciate any help that anyone could send my way. Have a great week. Tom Pier From contact <@t> excaliburpathology.com Tue Aug 23 09:57:41 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Automated tissue processor Message-ID: <20050823145742.66521.qmail@web50308.mail.yahoo.com> Just to clarify about the EMS tissue processor for plastics. It is for plastics that need high heat for polymerization and may not be suitable for methacrylates. From RitaR <@t> lexhealth.org Tue Aug 23 10:26:02 2005 From: RitaR <@t> lexhealth.org (Rita Riddle) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] PAS-F Message-ID: I am sure someone has this procedure. I have seen it somewhere , but can not find it. I need a PAS-F using Chromic Acid instead of Periodic Acid. Hope you can help. I am not talking about the Gridley stain. Thanks for you help . Rita Riddle HT(ASCP)PA Lead Histology Tech Lexington Medical Center West Columbia, SC Telephone 803-791-2881 Fax 803-791-2331 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From ree3 <@t> leicester.ac.uk Tue Aug 23 10:31:56 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] FERROPORTIN ANTIBODY Message-ID: I am after an antibody to ferroportin that works on rodent paraffin processed tissues..... Thanks Richard Edwards MRC TOXICOLOGY UNIT..U.K..... From chiggerson <@t> memhosp.com Tue Aug 23 11:39:41 2005 From: chiggerson <@t> memhosp.com (chiggerson@memhosp.com) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Hospital fetus disposition survey In-Reply-To: Message-ID: Histonetters, We are evaluating our current practices for fetus disposal at our facility and would like to know how both Catholic and non-Catholic hospitals handle fetus disposition at their facilities. I am interested in practices for both fetal loss greater than 20 weeks gestation AND less than 20 weeks gestation (products of conception). Please include religious affiliation, if any, of your institution. 1. Are parents given the option to have the hospital handle the disposition for the remains at < 20 wks, >20 weeks, or both? 2. If parents opt for hospital disposition, is it handled on-site, or contracted by the hospital off-site (with a funeral home)? If off-site, is the patient charged for disposition? 3. Method of disposition? Cremation? Burial? Other? 4. Are remains collected/disposed of separately, or co-mingled with other tissues? 5. Is disposal method dictated by State Law? Please include state. Any additional information you can give me about your practices would be helpful. Thank you in advance for your help. Cindy From llewllew <@t> shaw.ca Tue Aug 23 11:40:43 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] PAS-F References: Message-ID: <001401c5a801$68088670$7e034246@yourlk4rlmsu> Do you mean the PAFS (periodic acid fluorescent Schiff)? If so the procedure is on StainsFile (http://stainsfile.info). Select staining methods then look in the methods index. Bryan Llewellyn ----- Original Message ----- From: "Rita Riddle" To: Sent: Tuesday, August 23, 2005 8:26 AM Subject: [Histonet] PAS-F >I am sure someone has this procedure. I have seen it somewhere , but can > not find it. I need a PAS-F using Chromic Acid instead of Periodic Acid. > Hope you can help. I am not talking about the Gridley stain. Thanks for > you help . > > Rita Riddle HT(ASCP)PA > Lead Histology Tech > Lexington Medical Center > West Columbia, SC > Telephone 803-791-2881 > Fax 803-791-2331 > > _________________________________ > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of > its > attachments, please be advised that you have received this email in error > and that any use, dissemination, distribution, forwarding, printing, or > copying of this email or any attached files is strictly prohibited. If you > have received this email in error, please immediately purge it and all > attachments and notify the sender by reply email or contact the sender at > the number listed above if one is provided. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jqb7 <@t> cdc.gov Tue Aug 23 11:49:36 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] PAS-F Message-ID: Or PAS-F, PAS for fungal elements? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: Tuesday, August 23, 2005 12:41 PM To: Histonet Subject: Re: [Histonet] PAS-F Do you mean the PAFS (periodic acid fluorescent Schiff)? If so the procedure is on StainsFile (http://stainsfile.info). Select staining methods then look in the methods index. Bryan Llewellyn ----- Original Message ----- From: "Rita Riddle" To: Sent: Tuesday, August 23, 2005 8:26 AM Subject: [Histonet] PAS-F >I am sure someone has this procedure. I have seen it somewhere , but can > not find it. I need a PAS-F using Chromic Acid instead of Periodic Acid. > Hope you can help. I am not talking about the Gridley stain. Thanks for > you help . > > Rita Riddle HT(ASCP)PA > Lead Histology Tech > Lexington Medical Center > West Columbia, SC > Telephone 803-791-2881 > Fax 803-791-2331 > > _________________________________ > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of > its > attachments, please be advised that you have received this email in error > and that any use, dissemination, distribution, forwarding, printing, or > copying of this email or any attached files is strictly prohibited. If you > have received this email in error, please immediately purge it and all > attachments and notify the sender by reply email or contact the sender at > the number listed above if one is provided. > _______________________________________________ > Histonet mailing list > Histonet@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 sluhisto <@t> yahoo.com Tue Aug 23 11:53:50 2005 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Kappa Lambda on B5 fixed decaled BM biopsies Message-ID: <20050823165350.41911.qmail@web51006.mail.yahoo.com> Hello All: We have, for some time now, had issues with our bone marrow core biopsies and staining with Kappa and Lambda. Our cores are B5 fixed for 2 hours then decaled in RDO for one hour. They are processed overnight on the tissue processor and embedded the next morning. If anyone has any other protocols or more specifically could share your handling schedules and K/L protocols, I would be forever grateful. I am at a dead end and have a pathologist very upset that I can not get this "fixed". Thanks always for your help. Susan __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From gcallis <@t> montana.edu Tue Aug 23 12:00:19 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] PAS-F In-Reply-To: References: Message-ID: <6.0.0.22.1.20050823105453.01b32e68@gemini.msu.montana.edu> Easy, simply replace the periodic acid with 4% chromic acid (chromium trioxide is the dry chemical). This is the same reagent used in a GMS stain, and you do the same timing for treatment of section, rinse as you would for GMS then proceed to Schiffs reagent. Do not drain dump Chromic acid, it should be collected due to chromium ions. WE only use our chromic acid for a week, the dispose of it. At 09:26 AM 8/23/2005, you wrote: >I am sure someone has this procedure. I have seen it somewhere , but can >not find it. I need a PAS-F using Chromic Acid instead of Periodic Acid. >Hope you can help. I am not talking about the Gridley stain. Thanks for >you help . > >Rita Riddle HT(ASCP)PA >Lead Histology Tech >Lexington Medical Center >West Columbia, SC >Telephone 803-791-2881 >Fax 803-791-2331 > >_________________________________ >This email and any files transmitted with it may contain PRIVILEGED or >CONFIDENTIAL information and may be read or used only by the intended >recipient. If you are not the intended recipient of the email or any of its >attachments, please be advised that you have received this email in error >and that any use, dissemination, distribution, forwarding, printing, or >copying of this email or any attached files is strictly prohibited. If you >have received this email in error, please immediately purge it and all >attachments and notify the sender by reply email or contact the sender at >the number listed above if one is provided. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From weneng2004 <@t> yahoo.com Tue Aug 23 12:12:14 2005 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] How to wipe off omission oil from slides? Message-ID: <20050823171214.43420.qmail@web53407.mail.yahoo.com> Hello, I found our way wiping off omission oil after reading under microscope works not good enough. I want to hear how other people do. What solution you use? Thanks, Wen __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From TillRenee <@t> uams.edu Tue Aug 23 12:26:06 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] histostain Message-ID: Has anyone used the Histostain-Plus or Histostain-SP kits from Zymed on animal tissues? I know the Histomouse kit will work, but I need more bulk than 50 slides, but less price than what the 250 HIstomouse costs. I realize that there could be the background problem, I just want to know if it will work at all. I need some thing relatively cheap for a researcher with not a lot of money Renee' Till, HT From pruegg <@t> ihctech.net Tue Aug 23 12:45:17 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] How to wipe off omission oil from slides? In-Reply-To: <20050823171214.43420.qmail@web53407.mail.yahoo.com> Message-ID: <200508231745.j7NHj9nQ023777@chip.viawest.net> I use a wipe dipped in 70% alcohol to wipe off immersion oil from the lens and from the slide. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of wen eng Sent: Tuesday, August 23, 2005 10:12 AM To: histonet Subject: [Histonet] How to wipe off omission oil from slides? Hello, I found our way wiping off omission oil after reading under microscope works not good enough. I want to hear how other people do. What solution you use? Thanks, Wen __________________________________________________ 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 Nancy.Temple <@t> ssfhs.org Tue Aug 23 12:53:13 2005 From: Nancy.Temple <@t> ssfhs.org (Temple Nancy) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Re: Disposition of fetal remains Message-ID: Here at St. Francis Hospital in Indianapolis (Catholic hospital), fetal remains over 20 weeks are required by state law to be buried by parents. Fetal remains under 20 weeks may be buried, if parents choose. They must make arrangements with funeral home, and remains are released only to funeral home. Other fetal remains (POC specimens) are bagged together and sent to a funeral home periodically for cremation. Once a year, there is a memorial service at the funeral home, and any of the parents that want may attend. The hospital has a perinatal bereavement coordinator that handles communication with parents, etc. Nancy Temple HT(ASCP) Histology Supervisor St. Francis Hospital Indianapolis, In From pruegg <@t> ihctech.net Tue Aug 23 12:57:57 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] automated tissue processor for plastics In-Reply-To: <20050823141622.52842.qmail@web50308.mail.yahoo.com> Message-ID: <200508231757.j7NHvmnQ027786@chip.viawest.net> Years ago I used a carosel tissue processor that was for EM to process GMA infiltrated bone, but the reagent vials were very small limiting the size tissue you could process, I forget what it was called. It was actually small enough to put in a large fridge, we used it in a cold room. It did not accommondate the large undecalcified bones I did but it was pretty good for bone marrow biopsies taken with a jamshidi 8 gauge needle. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Pierce Sent: Tuesday, August 23, 2005 7:16 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] automated tissue processor for plastics EMS has a processor that will take specimens all the way to 100% resin. Here is the link: http://www.emsdiasum.com/microscopy/products/equipment/tissue_processor.aspx ?mm=19 Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From daniel.pirici <@t> ua.ac.be Tue Aug 23 13:09:08 2005 From: daniel.pirici <@t> ua.ac.be (Daniel Pirici) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] MAB 328 Message-ID: Has anybody had more experiences with Chemicon's MAB 328 (oligo marker) on paraffin sections ? Thanks, Daniel -- No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.338 / Virus Database: 267.10.14/79 - Release Date: 22-Aug-05 From brett_connolly <@t> merck.com Tue Aug 23 13:17:18 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Aurora A & B protocols anyone? Message-ID: <355C35514FEAC9458F75947F5270974D0795C5@usctmx1103.merck.com> I have been struggling with inconsistent results using these 2 Abcam antibodies recently on some formalin fixed tissues. I have ordered additional Ab's from some other suppliers and thought I'd throw out a plea for protocols in case anyone has a good one. Thanks, Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From ROrr <@t> enh.org Tue Aug 23 13:30:30 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] NSH Get together Message-ID: Hi Everyone, What do you think about a very very very informal meet and greet for Histonetters at NSH? I have gotten very useful responses and hopefully have been helpful with some of mine. I would appreciate putting some faces with some names. I'll just throw out a date, how about Monday Sept 12, around 6pm? Before the Thermo party...:-) If I get some interested responses maybe we can figure out a place. I'll spring for some name tags and let's see what happens! Please respond to me separately, and I'll share the general concensus. Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 From emry <@t> u.washington.edu Tue Aug 23 13:32:31 2005 From: emry <@t> u.washington.edu (Trisha Emry) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Seattle Microtome Repair Message-ID: Can anyone recommend a local repair service? Thanks, Trisha U of WA Seattle From Rcartun <@t> harthosp.org Tue Aug 23 13:45:17 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Kappa Lambda on B5 fixed decaled BM biopsies Message-ID: Number one - get rid of B5; use formalin! Number two - you need to use different antibody concentrations and retrieval methods to see light chain-restricted immunoreactivity in formalin-fixed, paraffin-embedded HemePath specimens. We have been moderately successful using a 96 degree waterbath for 40 minutes for retrieval; however, with lymphomas with abundant cytoplasmic immunoglobulin this method may be too sensitive and you may have to turn to an enzyme digestion (we use pepsin). We also have two different dilutions for both kappa and lambda. If the first attempt shows too much background, we will then repeat the studies using lower concentrations of antibody. Everyone's tissue is different, but this has worked for us. See our article, "Immunohistochemical detection of immunoglobulin light chain expression in ........" that appeared in Applied Immunohistochemistry & Molecular Morphology back in 2002 (volume 10/number 3). 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 >>> Histology SLU 08/23/05 12:53PM >>> Hello All: We have, for some time now, had issues with our bone marrow core biopsies and staining with Kappa and Lambda. Our cores are B5 fixed for 2 hours then decaled in RDO for one hour. They are processed overnight on the tissue processor and embedded the next morning. If anyone has any other protocols or more specifically could share your handling schedules and K/L protocols, I would be forever grateful. I am at a dead end and have a pathologist very upset that I can not get this "fixed". Thanks always for your help. Susan __________________________________________________ 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 ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From lance.erickson <@t> ihc.com Tue Aug 23 13:46:55 2005 From: lance.erickson <@t> ihc.com (Lance Erickson) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Hospital fetus disposition survey Message-ID: Here at Primary Children's Medical Center in Salt Lake City, UT (Does not maintain a religious affiliation although was set up originally by the LDS church and later sold to a non-profit hospital group) the parents are given the option of disposition <20 weeks and >20 weeks. As a children's hospital we do not have a labor and delivery department so all the cases we see are referral autopsies from outside institutions. Disposition is only performed by funeral homes. We are contracted with a local funeral home for cremation of those cases without disposition instructions or for those parents that elect for hospital disposition. The parents may request the ashes from the funeral home. It is our policy to not extend any charges to the patient/families for any postmortem work (this type of disposition becomes part of the autopsy service and is paid by the institution). Remains are handled individually and are not placed with any other tissues. The requirements of the cases that are >20 weeks are dictated by state law (so is the transportation of the remains anywhere from a referring institution). Please send all responses to the Histonet as we are also interested in this topic. Lance Erickson Anatomic Pathology Supervisor PCMC >>> 08/23/05 10:39 AM >>> Histonetters, We are evaluating our current practices for fetus disposal at our facility and would like to know how both Catholic and non-Catholic hospitals handle fetus disposition at their facilities. I am interested in practices for both fetal loss greater than 20 weeks gestation AND less than 20 weeks gestation (products of conception). Please include religious affiliation, if any, of your institution. 1. Are parents given the option to have the hospital handle the disposition for the remains at < 20 wks, >20 weeks, or both? 2. If parents opt for hospital disposition, is it handled on-site, or contracted by the hospital off-site (with a funeral home)? If off-site, is the patient charged for disposition? 3. Method of disposition? Cremation? Burial? Other? 4. Are remains collected/disposed of separately, or co-mingled with other tissues? 5. Is disposal method dictated by State Law? Please include state. Any additional information you can give me about your practices would be helpful. Thank you in advance for your help. Cindy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jtrynak <@t> hotmail.com Tue Aug 23 13:57:57 2005 From: jtrynak <@t> hotmail.com (John Rynak) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Wanted: Histoology Instrumentation Sales Professionals Message-ID: SciGenium's Client is experiencing tremendous growth within their North American operations. This growth is due to the recent launch of their FDA approved automated histology instrumentation system, and the growing antibody and reagent consumable product line for the histology market . Due to the new launch of these products They are currently looking for Senior Account Representatives, within the following regions Mid-Atlantic, Southwest, West Coast areas. The Ideal candidate will have had 3+ years of experience selling capital equipment and associated consumables into a clinical diagnostics laboratory. This person will have had prior experience selling to end user or laboratory manager ideally within the histopathology sector,and a BS/MS in a life science discipline. This position requires the ability to travel 75% of the time in a multistate region. Interested parties should forward a resume to [1]resume@scigenium.com SciGenium PO 380916 Cambridge, MA 02238 References 1. mailto:resume@scigenium.com From d.gregg <@t> juno.com Tue Aug 23 14:02:32 2005 From: d.gregg <@t> juno.com (d.gregg@juno.com) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Research histotech/virology technician opening Message-ID: <20050823.150233.2356.0.d.gregg@juno.com> I have a position open at Plum Island Animal Disease Center on Plum Island off of the East end of Long Island. It is research on foreign animal diseases-mostly pathogenesis work. The techniques include immunohistochemistry, virology, some paraffin, more cryosectioning. we also do laser confocal, laser dissection and electron microscopy. It is IA small unit supporting one veterinary pathologist and several post-docs. It is mainly a one person histo lab with great a variety of work. The area is very scenic and a resort destination, surrounded by water, and the job includes two boat rides each day to and from Plum Island, either to Long Island (10 minutes) or Connecticut (40 min.). Also included is laboratory cloths and one shower a day going out of the BL-3 facility. Most work is on non-zoonotic viruses such as foot-and-mouth disease and classical swine fever. The position has a wide grade range from GS 5-8 with regional differential cost of living pay added. If interested, please contact me directly. The position can be filled as a fellowship appointment or as a permanent full time employee in ARS, USDA. Douglas Gregg DVM, PhD Veterinary pathologist From dobbin <@t> upei.ca Tue Aug 23 15:05:59 2005 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] NSH Get together In-Reply-To: Message-ID: <430B4968.2901.17774D7@localhost> Hi Rebecca, I would think the most appropriate place to meet would be at the Histonet booth in the registration area. It will be called the "Histonet Outpost" and a few of us have done some work to pull this together for exactly the reasons to which you refer. :-) Have fun there. I'll have to meet you next time! Greg Date sent: Tue, 23 Aug 2005 13:30:30 -0500 From: "Orr, Rebecca" To: Subject: [Histonet] NSH Get together > > > Hi Everyone, > > What do you think about a very very very informal meet and greet for > Histonetters at NSH? > > > I have gotten very useful responses and hopefully have been helpful > with some of mine. > > I would appreciate putting some faces with some names. > > > > I'll just throw out a date, how about Monday Sept 12, around 6pm? > Before the Thermo party...:-) > > If I get some interested responses maybe we can figure out a place. > > > > I'll spring for some name tags and let's see what happens! > > Please respond to me separately, and I'll share the general concensus. > > > > Becky Orr, CLA HT (ASCP) > > IHC Lead > > Evanston Northwestern Healthcare > > ph: 847-570-2771 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Research Technologist, Pathology Lab, Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From JNocito <@t> Pathreflab.com Tue Aug 23 14:14:31 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] NSH Get together In-Reply-To: Message-ID: count me in 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 Orr, Rebecca Sent: Tuesday, August 23, 2005 1:31 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] NSH Get together Hi Everyone, What do you think about a very very very informal meet and greet for Histonetters at NSH? I have gotten very useful responses and hopefully have been helpful with some of mine. I would appreciate putting some faces with some names. I'll just throw out a date, how about Monday Sept 12, around 6pm? Before the Thermo party...:-) If I get some interested responses maybe we can figure out a place. I'll spring for some name tags and let's see what happens! Please respond to me separately, and I'll share the general concensus. Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Aug 23 14:46:18 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Seattle Microtome Repair In-Reply-To: References: Message-ID: <6.0.0.22.1.20050823134529.01b56288@gemini.msu.montana.edu> If it is a Leica, call Bartells and Stout, they repair in house at their Seattle area location. At 12:32 PM 8/23/2005, you wrote: >Can anyone recommend a local repair service? >Thanks, >Trisha >U of WA >Seattle > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From pex0220 <@t> yahoo.com.cn Tue Aug 23 14:51:44 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] decalcification Message-ID: <20050823195144.1512.qmail@web15509.mail.cnb.yahoo.com> Hello,all Does cartilage need to be decalcified? Thank you! Guofeng --------------------------------- DO YOU YAHOO!? ÑÅ»¢Ãâ·ÑGÓÊÏ䣭ÖйúµÚÒ»¾øÎÞÀ¬»øÓʼþɧÈų¬´óÓÊÏä From PMonfils <@t> Lifespan.org Tue Aug 23 15:05:20 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:29 2005 Subject: FW: [Histonet] decalcification Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175BC@lsexch.lsmaster.lifespan.org> Cartilage does not require decalcification, since it does not contain calcium deposits. Neither does heavily keratinized material like nails. Both substances however will benefit from extended processing time. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of pex > Sent: Tuesday, August 23, 2005 12:51 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] decalcification > > Hello,all > > Does cartilage need to be decalcified? > > Thank you! > > Guofeng > > > > > --------------------------------- > DO YOU YAHOO!? > ????G??-???????????????? > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gcallis <@t> montana.edu Tue Aug 23 15:33:08 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Cartilage and decalcification Message-ID: <6.0.0.22.1.20050823141305.01b78f50@gemini.msu.montana.edu> >Articular cartilage located over the ends of bones in joints does not need >decalcification unless you pull it off adjacent to the bone and bone >fragments remain attached to the cartilage. Cartilagenous matrix in >epiphysis of developing long bones will calcify hence there is such a >thing as calcified cartilage. Young individuals will have calcifying >cartilage present in the areas of growth plate between epiphysis and >metaphysis. You will see this as columns of developing bone where >cartilage matrix is calcifying. >A good histology textbook, not one that is for HISTOTECHNOLOGY, but to >teach tissue morphology and how organs form, etc will explain bone bone >formation from prenatal endochondral bone formation to a mature, adult >individual. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From RCazares <@t> schosp.org Tue Aug 23 15:38:21 2005 From: RCazares <@t> schosp.org (Cazares, Ruth) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Re: NSH get together Message-ID: <913FAC2B773C19488E26AE6572180FA502C6F7D6@exch01.schosp.org> Will there be libations (of the ETOH kind)? That always makes for a fun time!! Ruth Cazares Swedish Covenant Hospital Chicago, IL *** Confidentiality Statement *** This e-mail is intended only for the use of the individual or entity to which it is addressed and may contain information that is privileged and confidential. If the reader of this message is not the intended recipient, please notify the sender immediately by replying to this message and then delete it from your system. Any review, dissemination, distribution, or reproduction of this message by unintended recipients is strictly prohibited and may be subject to legal restriction. Thank you for your cooperation. From katherine-walters <@t> uiowa.edu Tue Aug 23 16:52:54 2005 From: katherine-walters <@t> uiowa.edu (Walters, Katherine S) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] puzzling morphology Message-ID: I am once again in need of some collective wisdom from the experts. I have some paraffin embedded lung tissue in which the bronchial endothelium cells exhibit "spaces" (around the nucleus) in the cytoplasm in the H&E 5 micron section. These were fixed with zinc formalin. Does anyone know what would cause this absence of tissue? Thanks again for your wisdom, Kathy From rod.coombe <@t> imvs.sa.gov.au Tue Aug 23 17:04:54 2005 From: rod.coombe <@t> imvs.sa.gov.au (Rod Coombe) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Cassette/slide writers Message-ID: <000201c5a82e$b1257070$d86d140a@ITP36079> I would be very interested in feedback on cassette and slide writers. Which ones are performing and how it is interfaced to your laboratory information system. Any information would be most appreciated. Rod Coombe Manager Tissue Pathology IMVS Frome Road, Adelaide Phone 61 8 8222 3201 / 0401 120808 Fax 61 8 8222 3204 From gcallis <@t> montana.edu Tue Aug 23 17:06:06 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Using chromic acid instead of periodic acid with Schiffs - for fungus staining ONLY! Message-ID: <6.0.0.22.1.20050823150922.01b08c70@gemini.msu.montana.edu> Histonetters, I hope I didn't confuse people with my answer about PAS-F (what the F meant escapes me??? for fungus, fluorescence?) where the person was asking about using chromic acid instead of periodic acid i.e in PAS. Theoretically, one could not call this PAS - maybe chromic acid-Schiff or CAS?? The key word was the mention of Gridley staining, a method for fungus staining, and I assumed this person wanted to use a chromic acid oxidizer followed by Schiffs reagent for that purpose i.e fungus stain. Whatever you do, do NOT use chromic acid for a standard PAS stain if staining for mucosubstances, glycogen, basement membranes or other components that are PAS positive. Chromic acid is a much stronger oxidizer than periodic acid, and will over-oxidize these components to the point of them NOT staining after Schiffs application - not a good idea. However chromic acid with Schiffs does work for fungus staining. Freida Carson et al wrote a publication on false negative fungus staining by using Periodic acid -Schiffs reagent, and mentioned using chromic acid in place of periodic acid. This can be found in J of Histotechnology, and an excellent bit of information. Chromic acid oxidation with Schiffs reagents is reserved for those who do not want use periodic acid as the oxidizing agent for fungus staining. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From rod.coombe <@t> imvs.sa.gov.au Tue Aug 23 17:25:33 2005 From: rod.coombe <@t> imvs.sa.gov.au (Rod Coombe) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Accession numbers and slide, block storage Message-ID: <000301c5a831$93d6f810$d86d140a@ITP36079> Has anybody had experience with random numbering systems. Currently we use a yearly sequential numbering system which reverts back to a set number on the 1st January each year. A few years ago we trialed random numbering (which was entered onto the lab information system) while a second sequential number was used for internal filing. This was a disaster, trying to cope with two numbers and working out where slides were filed and was subsequently abandoned. Does any lab have experience using 1D or 2D bar codes for tracking blocks and slides. If so is it only possible using sequential numbers or can it be accomplished with random numbers. Your experiences would be welcomed. Rod Coombe Manager Tissue Pathology IMVS Frome Road, Adelaide Phone (08) 8222 3201 / 0401 120808 Fax (08) 82223204 From Xudong_Cao <@t> brown.edu Tue Aug 23 18:42:12 2005 From: Xudong_Cao <@t> brown.edu (Cao, Xudong) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] PDGF retrovirus packaging cell line Message-ID: <1DA88DDC48CCC245A777599FDAEED6D002C6035D@MAIL1.AD.Brown.Edu> Hello all, anybody knows where I can get PDGF retrovirus packaging cell line? (e.g. where, who is the PI I need to contact) I know this is way off the topic...sorry. many thanks. cheers, Xudong Cao, Ph.D. Assistant Professor From llewllew <@t> shaw.ca Tue Aug 23 18:29:34 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Using chromic acid instead of periodic acid with Schiffs - for fungus staining ONLY! References: <6.0.0.22.1.20050823150922.01b08c70@gemini.msu.montana.edu> Message-ID: <000801c5a83a$85529ac0$7e034246@yourlk4rlmsu> I think the person wanted the details for Bauer's stain, or CAS (chromic acid Schiff). It was originally published for glycogen, I believe, but PAS is better for that. The method is simple. Oxidise with 1% chromium trioxide in water for 1 hour (or 5% for 10 minutes). Bleach with 5% metabisulphite and wash. Schiff's reagent 20 minutes or so. Finish off as you would a PAS. Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, August 23, 2005 3:06 PM Subject: [Histonet] Using chromic acid instead of periodic acid with Schiffs - for fungus staining ONLY! > Histonetters, > > I hope I didn't confuse people with my answer about PAS-F (what the F > meant escapes me??? for fungus, fluorescence?) where the person was asking > about using chromic acid instead of periodic acid i.e in PAS. > Theoretically, one could not call this PAS - maybe chromic acid-Schiff or > CAS?? The key word was the mention of Gridley staining, a method for > fungus staining, and I assumed this person wanted to use a chromic acid > oxidizer followed by Schiffs reagent for that purpose i.e fungus stain. > > Whatever you do, do NOT use chromic acid for a standard PAS stain if > staining for mucosubstances, glycogen, basement membranes or other > components that are PAS positive. Chromic acid is a much stronger > oxidizer than periodic acid, and will over-oxidize these components to the > point of them NOT staining after Schiffs application - not a good idea. > However chromic acid with Schiffs does work for fungus staining. Freida > Carson et al wrote a publication on false negative fungus staining by > using Periodic acid -Schiffs reagent, and mentioned using chromic acid in > place of periodic acid. This can be found in J of Histotechnology, and an > excellent bit of information. > > Chromic acid oxidation with Schiffs reagents is reserved for those who do > not want use periodic acid as the oxidizing agent for fungus staining. > > Gayle Callis > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From AnthonyH <@t> chw.edu.au Tue Aug 23 19:31:32 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Hospital fetus disposition survey Message-ID: Cindy, The laws in Australia are different to elsewhere but in summery: Stillborns greater than 20 weeks need to be buried/cremated. Under 20 weeks to be disposed of a/c to families wishes. At our hospital, unless otherwise requested, we keep the fetuses for 6 months prior to having them cremated by a local crematorium. The ashes are then sprinkled on the rose garden at the hospital. The crematorium does this as a big favour and we are in their debt. This procedure only applies to those fetuses that have had a postmortem. The parents permission is always obtained. Hope this helps Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of chiggerson@memhosp.com Sent: Wednesday, 24 August 2005 2:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Hospital fetus disposition survey Histonetters, We are evaluating our current practices for fetus disposal at our facility and would like to know how both Catholic and non-Catholic hospitals handle fetus disposition at their facilities. I am interested in practices for both fetal loss greater than 20 weeks gestation AND less than 20 weeks gestation (products of conception). Please include religious affiliation, if any, of your institution. 1. Are parents given the option to have the hospital handle the disposition for the remains at < 20 wks, >20 weeks, or both? 2. If parents opt for hospital disposition, is it handled on-site, or contracted by the hospital off-site (with a funeral home)? If off-site, is the patient charged for disposition? 3. Method of disposition? Cremation? Burial? Other? 4. Are remains collected/disposed of separately, or co-mingled with other tissues? 5. Is disposal method dictated by State Law? Please include state. Any additional information you can give me about your practices would be helpful. Thank you in advance for your help. Cindy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From bhewlett <@t> cogeco.ca Tue Aug 23 21:32:39 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Using chromic acid instead of periodic acid with Schiffs - for fungus staining ONLY! References: <6.0.0.22.1.20050823150922.01b08c70@gemini.msu.montana.edu> Message-ID: <001701c5a854$19114a90$6400a8c0@mainbox> What Rita was asking about PAS-F, I don't know (what does the -F stand for Rita?). However, the Bauer reaction (chromic acid oxidation-Schiff) in 1933 was the first ever use in histochemistry of a glycol to aldehyde oxidation reaction. This produces a positive Schiff result in a variety of mucosubstances, including cellulose, starch and (sorry Gayle) GLYCOGEN! It also gives a positive result with fungal cell walls. In fact the Bauer-Schiff was for many years the method of choice for the demonstration of glycogen and other mucosubstances. As Gayle correctly pointed out, chromic acid further oxidizes the aldehydes , probably to carboxyl groups and then to carbon dioxide. Mucosubstances with a high glycol density (glycogen, starch, fungal cell walls) are more resistant to this effect. The resulting Schiff reaction is somewhat paler than a PAS, how much paler depends on oxidation time, but still positive. Gomori (1946) used chromic acid oxidation in his method for glycogen and mucin(sic). He detected the resulting aldehydes with hexamine-silver. Grocott (1955) optimized the Gomori technique for fungal cell walls. The Gridley stain (1953) was just a Bauer-Schiff/aldehyde fuchsine combination stain. Periodic oxidation-Schiff was introduced by both Hotchkiss and McManus in 1945-46. Periodic acid does not further oxidize the aldehydes produced (at least not in any reasonable time frame) and rapidly became the oxidation-Schiff method of choice. Bryan ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, August 23, 2005 6:06 PM Subject: [Histonet] Using chromic acid instead of periodic acid with Schiffs - for fungus staining ONLY! > Histonetters, > > I hope I didn't confuse people with my answer about PAS-F (what the F > meant escapes me??? for fungus, fluorescence?) where the person was asking > about using chromic acid instead of periodic acid i.e in PAS. > Theoretically, one could not call this PAS - maybe chromic acid-Schiff or > CAS?? The key word was the mention of Gridley staining, a method for > fungus staining, and I assumed this person wanted to use a chromic acid > oxidizer followed by Schiffs reagent for that purpose i.e fungus stain. > > Whatever you do, do NOT use chromic acid for a standard PAS stain if > staining for mucosubstances, glycogen, basement membranes or other > components that are PAS positive. Chromic acid is a much stronger > oxidizer than periodic acid, and will over-oxidize these components to the > point of them NOT staining after Schiffs application - not a good idea. > However chromic acid with Schiffs does work for fungus staining. Freida > Carson et al wrote a publication on false negative fungus staining by > using Periodic acid -Schiffs reagent, and mentioned using chromic acid in > place of periodic acid. This can be found in J of Histotechnology, and an > excellent bit of information. > > Chromic acid oxidation with Schiffs reagents is reserved for those who do > not want use periodic acid as the oxidizing agent for fungus staining. > > Gayle Callis > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jkiernan <@t> uwo.ca Wed Aug 24 00:53:07 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Hospital fetus disposition survey References: Message-ID: <430C0B43.71A17DA8@uwo.ca> Dear Tony, The Australian regulations reflect your country's ethical standards, which are probably the highest in the world when it comes to recognizing human embryos, human fetuses and demented adults as human beings. How do Australian abortionists dispose of their spoils? In Canada, paid-for abortuses go through a kitchen-sink garbage masher and then into the drains. Abortion is legal in Canada right up to 40 weeks, with the intrauterine decapitation method. Any Canadian baby can be killed just before birth at the whim of its mother. The mother must not be asked for a reason, but she has to pay the abortionist's fee. John Kiernan London, Canada ------------------------- Tony Henwood wrote: > > Cindy, > > The laws in Australia are different to elsewhere but in summery: > > Stillborns greater than 20 weeks need to be buried/cremated. > Under 20 weeks to be disposed of a/c to families wishes. At our hospital, > unless otherwise requested, we keep the fetuses for 6 months prior to having > them cremated by a local crematorium. The ashes are then sprinkled on the > rose garden at the hospital. The crematorium does this as a big favour and > we are in their debt. This procedure only applies to those fetuses that have > had a postmortem. The parents permission is always obtained. > > Hope this helps > > Regards > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager & Senior Scientist > The Children's Hospital at Westmead, > Locked Bag 4001, Westmead, 2145, AUSTRALIA. > Tel: 612 9845 3306 > Fax: 612 9845 3318 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > chiggerson@memhosp.com > Sent: Wednesday, 24 August 2005 2:40 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Hospital fetus disposition survey > > Histonetters, > > We are evaluating our current practices for fetus disposal at our facility > and would like to know how both Catholic and non-Catholic hospitals handle > fetus disposition at their facilities. I am interested in practices for > both fetal loss greater than 20 weeks gestation AND less than 20 weeks > gestation (products of conception). > > Please include religious affiliation, if any, of your institution. > > 1. Are parents given the option to have the hospital handle the > disposition for the remains at < 20 wks, >20 weeks, or both? > > 2. If parents opt for hospital disposition, is it handled on-site, or > contracted by the hospital off-site (with a funeral home)? If off-site, > is the patient charged for disposition? > > 3. Method of disposition? Cremation? Burial? Other? > > 4. Are remains collected/disposed of separately, or co-mingled with > other tissues? > > 5. Is disposal method dictated by State Law? Please include state. > > Any additional information you can give me about your practices would be > helpful. > > Thank you in advance for your help. > > Cindy > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ********************************************************************** > This email and any files transmitted with it are confidential and > intended solely for the use of the individual or entity to whom they > are addressed. If you are not the intended recipient, please > delete it and notify the sender. > > Views expressed in this message and any attachments are those > of the individual sender, and are not necessarily the views of The > Children's Hospital at Westmead > > This footnote also confirms that this email message has been > virus scanned and although no computer viruses were detected, > the Childrens Hospital at Westmead accepts no liability for any > consequential damage resulting from email containing computer > viruses. > ********************************************************************** > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Aug 24 02:38:53 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Hospital fetus disposition survey Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD500@elht-exch1.xelht.nhs.uk> In the UK Hospitals abortions are limited to I think 24 weeks and I think it's going to be reduced. Abortions can be carried out later if the Mother's life is at risk and usually the foetus is vacuumed out; we would never decapitate. All such foetuses are cremated or given to the parents for burial; whatever is there wish. I don't know what happens in the Clinics that offer these procedures; it would be of interest to know. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: 24 August 2005 06:53 To: Tony Henwood Cc: histonet@lists.utsouthwestern.edu; 'chiggerson@memhosp.com' Subject: Re: [Histonet] Hospital fetus disposition survey Dear Tony, The Australian regulations reflect your country's ethical standards, which are probably the highest in the world when it comes to recognizing human embryos, human fetuses and demented adults as human beings. How do Australian abortionists dispose of their spoils? In Canada, paid-for abortuses go through a kitchen-sink garbage masher and then into the drains. Abortion is legal in Canada right up to 40 weeks, with the intrauterine decapitation method. Any Canadian baby can be killed just before birth at the whim of its mother. The mother must not be asked for a reason, but she has to pay the abortionist's fee. John Kiernan London, Canada ------------------------- Tony Henwood wrote: > > Cindy, > > The laws in Australia are different to elsewhere but in summery: > > Stillborns greater than 20 weeks need to be buried/cremated. > Under 20 weeks to be disposed of a/c to families wishes. At our hospital, > unless otherwise requested, we keep the fetuses for 6 months prior to having > them cremated by a local crematorium. The ashes are then sprinkled on the > rose garden at the hospital. The crematorium does this as a big favour and > we are in their debt. This procedure only applies to those fetuses that have > had a postmortem. The parents permission is always obtained. > > Hope this helps > > Regards > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager & Senior Scientist > The Children's Hospital at Westmead, > Locked Bag 4001, Westmead, 2145, AUSTRALIA. > Tel: 612 9845 3306 > Fax: 612 9845 3318 > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > chiggerson@memhosp.com > Sent: Wednesday, 24 August 2005 2:40 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Hospital fetus disposition survey > > Histonetters, > > We are evaluating our current practices for fetus disposal at our facility > and would like to know how both Catholic and non-Catholic hospitals handle > fetus disposition at their facilities. I am interested in practices for > both fetal loss greater than 20 weeks gestation AND less than 20 weeks > gestation (products of conception). > > Please include religious affiliation, if any, of your institution. > > 1. Are parents given the option to have the hospital handle the > disposition for the remains at < 20 wks, >20 weeks, or both? > > 2. If parents opt for hospital disposition, is it handled on-site, or > contracted by the hospital off-site (with a funeral home)? If off-site, > is the patient charged for disposition? > > 3. Method of disposition? Cremation? Burial? Other? > > 4. Are remains collected/disposed of separately, or co-mingled with > other tissues? > > 5. Is disposal method dictated by State Law? Please include state. > > Any additional information you can give me about your practices would be > helpful. > > Thank you in advance for your help. > > Cindy > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ********************************************************************** > This email and any files transmitted with it are confidential and > intended solely for the use of the individual or entity to whom they > are addressed. If you are not the intended recipient, please > delete it and notify the sender. > > Views expressed in this message and any attachments are those > of the individual sender, and are not necessarily the views of The > Children's Hospital at Westmead > > This footnote also confirms that this email message has been > virus scanned and although no computer viruses were detected, > the Childrens Hospital at Westmead accepts no liability for any > consequential damage resulting from email containing computer > viruses. > ********************************************************************** > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Aug 24 02:48:19 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] puzzling morphology Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD501@elht-exch1.xelht.nhs.uk> It's not actually an absence of tissue is it? The spaces could be due to virus, like koilocytotic atypia. It could also be an artefact as you suggest but I can't figure what could cause perinuclear haloing as you suggest, I would have thought cellular changes would have affected the whole cell not just shrinking the nucleus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Walters, Katherine S Sent: 23 August 2005 22:53 To: histonet@pathology.swmed.edu Subject: [Histonet] puzzling morphology I am once again in need of some collective wisdom from the experts. I have some paraffin embedded lung tissue in which the bronchial endothelium cells exhibit "spaces" (around the nucleus) in the cytoplasm in the H&E 5 micron section. These were fixed with zinc formalin. Does anyone know what would cause this absence of tissue? Thanks again for your wisdom, Kathy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Paul.B.Atkinson <@t> rli.mbht.nhs.uk Wed Aug 24 04:18:44 2005 From: Paul.B.Atkinson <@t> rli.mbht.nhs.uk (Atkinson Paul B) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Michel's medium for skin biopsies Message-ID: We are considering using Michels's transport medium for skin biopsies in our laboratory. Have other laboratories found its use worthwhile? The original 1973 paper only mentions the effect on immunoglobulin staining, is it also effective in preserving complement antigenicity? Also how long does the tissue need to be in the fluid for the 'fixation' to occur? Many thanks Paul Atkinson DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From marshall <@t> cormack.uct.ac.za Wed Aug 24 05:26:53 2005 From: marshall <@t> cormack.uct.ac.za (Marshall) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] brain sections lifting Message-ID: <430C4B6D.46562CA0@cormack.uct.ac.za> Hi I have a student who is doing fluorescent immunochemistry and has problems with her sections lifting off her slides. We are using aptes coated slides but they are still lifting. I have told her to make sure the sections are as flat as possible when she picks them up from the waterbath but we still have a problem. She is also leaving them overnight at 60 degrees. Does anybody have any suggestions. I thought of using chrome gelatin but I am not sure if this is going to help. Thanks Sharon Marshall Dept. Human Biology University of Cape Town South Africa e-mail: marshall@cormack.uct.ac.za From vanann702 <@t> skmc.gov.ae Wed Aug 24 06:02:17 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] brain sections lifting Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E02F6D5E9@SKMCEMAIL.skmc.gov.ae> Hi Sharon This is Anne - I ran the Neuro lab at med school for many years and we met a few times. Perhaps you remember. Your subject line states 'brain sections lifting' I would leave them at 37 overnight instead of 60. save that for just prior to the dewax step brain washes off easily, more so than other tissue. thicker sections wash off easier than thinner ones. animal tissue washes off quicker than human. If these are thick, animal brains then you have a 'triple' dose of 'wash off' factor' if you still have trouble go across to the med school histo routine lab and find Patricia Papier - she's a whizz at this sort of thing. Tell her I send love. Cheers Annie in Arabia -----Original Message----- From: Marshall [mailto:marshall@cormack.uct.ac.za] Sent: Wednesday, August 24, 2005 2:27 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] brain sections lifting Hi I have a student who is doing fluorescent immunochemistry and has problems with her sections lifting off her slides. We are using aptes coated slides but they are still lifting. I have told her to make sure the sections are as flat as possible when she picks them up from the waterbath but we still have a problem. She is also leaving them overnight at 60 degrees. Does anybody have any suggestions. I thought of using chrome gelatin but I am not sure if this is going to help. Thanks Sharon Marshall Dept. Human Biology University of Cape Town South Africa e-mail: marshall@cormack.uct.ac.za _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jim.manavis <@t> imvs.sa.gov.au Wed Aug 24 06:52:11 2005 From: jim.manavis <@t> imvs.sa.gov.au (Jim Manavis) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Dopamine receptors Message-ID: <001a01c5a8a2$435db470$636c140a@itp36533> Has anyone experience with the Dopamine D1 and D2 receptors in formalin fixed paraffin embedded brain tissue. Cheers Jim From wasielewski.reinhard.von <@t> mh-hannover.de Wed Aug 24 07:22:07 2005 From: wasielewski.reinhard.von <@t> mh-hannover.de (wasielewski.reinhard.von@mh-hannover.de) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] VEGF and VEGF-R in FFPE mouse In-Reply-To: Message-ID: <430C828F.5831.9BCA1F5D@localhost> Dear Histonetters, may I ask your help to find out the best antibody and staining procedure for formalin- fixed, paraffin embedded mouse tissue for the markers VEGF and VEGFR ? Many thanks in advance - best regards Reinhard PD Dr. med. Reinhard von Wasielewski From Sarah_Mack <@t> urmc.rochester.edu Wed Aug 24 07:43:27 2005 From: Sarah_Mack <@t> urmc.rochester.edu (Mack, Sarah) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] VEGF and VEGF-R in FFPE mouse Message-ID: We use Santa Cruz VEGF antibody, catalog #SC-507. Sarah A Mack University of Rochester Cardiovascular Research Institute (CRI) 575 Elmwood Avenue, MRBX 2-11001-A Rochester, NY 14572 585-273-1914 > ---------- > From: wasielewski.reinhard.von@mh-hannover.de > Sent: Wednesday, August 24, 2005 8:22 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] VEGF and VEGF-R in FFPE mouse > > Dear Histonetters, > may I ask your help to find out the best antibody and staining procedure > for formalin- > fixed, paraffin embedded mouse tissue for the markers VEGF and VEGFR ? > > Many thanks in advance - best regards > > Reinhard > > > > PD Dr. med. Reinhard von Wasielewski > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Julie.Sanders <@t> med.va.gov Wed Aug 24 07:15:34 2005 From: Julie.Sanders <@t> med.va.gov (Julie.Sanders@med.va.gov) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] RE: Histonet Digest, Vol 21, Issue 29 Message-ID: <457381D92B01BD44B21CF37CC02EBDFD0292771D@vhacinexc2.v10.med.va.gov> This question is for folks who use the Leica autostainer. I'd like to see various H&E protocols for this stainer and see what everyone is using. I appreciate all responses. Thanks, Julie Sanders, BA, HT(ASCP) Supervisor, Anatomic Pathology VAMC, Cincinnati, Ohio From am458 <@t> cornell.edu Wed Aug 24 08:58:45 2005 From: am458 <@t> cornell.edu (Amanda MacFarlane) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Hospital fetus disposition survey In-Reply-To: <430C0B43.71A17DA8@uwo.ca> References: <430C0B43.71A17DA8@uwo.ca> Message-ID: <1250.128.253.96.73.1124891925.squirrel@128.253.96.73> In response to John Kiernan's reply: The question to which John Kiernan was replying is valid and deserves an objective response based on the specific questions asked. The Histonet should not be used as a forum for grandstanding, especially in regards to a highly controversial subject such as abortion, whether you share pro-life or pro-choice views. I found this response both unprofessional in its strong and uncalled for opinion, which is not even a direct response to the questions asked; it is also wrong in regards to abortions performed in Canada, in hospital or otherwise. In Canada, 90% of abortions are performed in the first trimester, and doctors do not perform abortions past 20 to 21 weeks except for health or genetic reasons. This is a professional forum, let's please keep our reponses as such and our personal opions to ourselves. Regards, Amanda MacFarlane, PhD Division of Nutritional Sciences Cornell University > Dear Tony, > > The Australian regulations reflect your country's > ethical standards, which are probably the highest in > the world when it comes to recognizing human embryos, > human fetuses and demented adults as human beings. > > How do Australian abortionists dispose of their > spoils? In Canada, paid-for abortuses go through > a kitchen-sink garbage masher and then into > the drains. Abortion is legal in Canada right up > to 40 weeks, with the intrauterine decapitation > method. Any Canadian baby can be killed just before > birth at the whim of its mother. The mother must > not be asked for a reason, but she has to pay > the abortionist's fee. > > John Kiernan > London, Canada > ------------------------- > Tony Henwood wrote: >> >> Cindy, >> >> The laws in Australia are different to elsewhere but in summery: >> >> Stillborns greater than 20 weeks need to be buried/cremated. >> Under 20 weeks to be disposed of a/c to families wishes. At our >> hospital, >> unless otherwise requested, we keep the fetuses for 6 months prior to >> having >> them cremated by a local crematorium. The ashes are then sprinkled on >> the >> rose garden at the hospital. The crematorium does this as a big favour >> and >> we are in their debt. This procedure only applies to those fetuses that >> have >> had a postmortem. The parents permission is always obtained. >> >> Hope this helps >> >> >> >> Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) >> Laboratory Manager & Senior Scientist >> The Children's Hospital at Westmead, >> Locked Bag 4001, Westmead, 2145, AUSTRALIA. >> Tel: 612 9845 3306 >> Fax: 612 9845 3318 >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >> chiggerson@memhosp.com >> Sent: Wednesday, 24 August 2005 2:40 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Hospital fetus disposition survey >> >> Histonetters, >> >> We are evaluating our current practices for fetus disposal at our >> facility >> and would like to know how both Catholic and non-Catholic hospitals >> handle >> fetus disposition at their facilities. I am interested in practices for >> both fetal loss greater than 20 weeks gestation AND less than 20 weeks >> gestation (products of conception). >> >> Please include religious affiliation, if any, of your institution. >> >> 1. Are parents given the option to have the hospital handle the >> disposition for the remains at < 20 wks, >20 weeks, or both? >> >> 2. If parents opt for hospital disposition, is it handled on-site, >> or >> contracted by the hospital off-site (with a funeral home)? If off-site, >> is the patient charged for disposition? >> >> 3. Method of disposition? Cremation? Burial? Other? >> >> 4. Are remains collected/disposed of separately, or co-mingled with >> other tissues? >> >> 5. Is disposal method dictated by State Law? Please include state. >> >> Any additional information you can give me about your practices would be >> helpful. >> >> Thank you in advance for your help. >> >> Cindy >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> ********************************************************************** >> This email and any files transmitted with it are confidential and >> intended solely for the use of the individual or entity to whom they >> are addressed. If you are not the intended recipient, please >> delete it and notify the sender. >> >> Views expressed in this message and any attachments are those >> of the individual sender, and are not necessarily the views of The >> Children's Hospital at Westmead >> >> This footnote also confirms that this email message has been >> virus scanned and although no computer viruses were detected, >> the Childrens Hospital at Westmead accepts no liability for any >> consequential damage resulting from email containing computer >> viruses. >> ********************************************************************** >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From vazquezr <@t> ohsu.edu Wed Aug 24 09:13:59 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Michel's medium for skin biopsies Message-ID: Paul, I believe Michels is a transport media, not a fixative media. You don't want the tissue fixed, if you are doing antibody testing on it, in the fresh tissue sense. I use Zeus Tissue Transport Media, my doctor seems to like it better. Robyn OHSU >>> "Atkinson Paul B" 8/24/2005 2:18 AM >>> We are considering using Michels's transport medium for skin biopsies in our laboratory. Have other laboratories found its use worthwhile? The original 1973 paper only mentions the effect on immunoglobulin staining, is it also effective in preserving complement antigenicity? Also how long does the tissue need to be in the fluid for the 'fixation' to occur? Many thanks Paul Atkinson DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Aug 24 09:20:39 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Thanks for info on Bauer reaction - chromic acid/Schiffs In-Reply-To: <001701c5a854$19114a90$6400a8c0@mainbox> References: <6.0.0.22.1.20050823150922.01b08c70@gemini.msu.montana.edu> <001701c5a854$19114a90$6400a8c0@mainbox> Message-ID: <6.0.0.22.1.20050824081115.01b4cea8@gemini.msu.montana.edu> Bryan, Thank you for the historical and technical info on this, I was not aware of the Bauer reaction, for glycogen (no apologies needed as it makes perfect sense!) - still learning after all these years. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) At 08:32 PM 8/23/2005, you wrote: >What Rita was asking about PAS-F, I don't know (what does the -F stand >for Rita?). >However, the Bauer reaction (chromic acid oxidation-Schiff) in 1933 >was the first ever use in histochemistry of a glycol to aldehyde oxidation >reaction. >This produces a positive Schiff result in a variety of mucosubstances, >including cellulose, starch and (sorry Gayle) GLYCOGEN! >It also gives a positive result with fungal cell walls. >In fact the Bauer-Schiff was for many years the method of choice for the >demonstration of glycogen and other mucosubstances. > >As Gayle correctly pointed out, chromic acid further oxidizes the >aldehydes , probably to carboxyl groups and then to carbon dioxide. >Mucosubstances with a high glycol density (glycogen, starch, fungal cell >walls) are more resistant to this effect. >The resulting Schiff reaction is somewhat paler than a PAS, how much paler >depends on oxidation time, but still positive. >Gomori (1946) used chromic acid oxidation in his method for glycogen and >mucin(sic). >He detected the resulting aldehydes with hexamine-silver. >Grocott (1955) optimized the Gomori technique for fungal cell walls. >The Gridley stain (1953) was just a Bauer-Schiff/aldehyde fuchsine >combination stain. >Periodic oxidation-Schiff was introduced by both Hotchkiss and McManus in >1945-46. >Periodic acid does not further oxidize the aldehydes produced (at least >not in any reasonable time frame) and rapidly became the oxidation-Schiff >method of choice. > >Bryan From mucram11 <@t> comcast.net Wed Aug 24 09:40:32 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:25:29 2005 Subject: [Histonet] Michel's medium for skin biopsies Message-ID: <082420051440.14304.430C86E00003FD4A000037E02200762194CECE030E9D0C9A03@comcast.net> Hi Paul, Michel's is a transport media not a fixative as Robyn indicated. It is similar to Zeus in some components. It really depends on what you need a fixative or something to move the tissue and preserve the antigenic sites during that process. The transport media are generally only good for holding 24 to 48 hours with most recommending 24 hours or less for EM. Good Luck, Pam Marcum -------------- Original message -------------- > Paul, > I believe Michels is a transport media, not a fixative media. You don't want > the tissue fixed, if you are doing antibody testing on it, in the fresh tissue > sense. I use Zeus Tissue Transport Media, my doctor seems to like it better. > > Robyn > OHSU > > >>> "Atkinson Paul B" 8/24/2005 2:18 AM >>> > > We are considering using Michels's transport medium for skin biopsies in our > laboratory. Have other laboratories found its use worthwhile? The original > 1973 paper only mentions the effect on immunoglobulin staining, is it also > effective in preserving complement antigenicity? Also how long does the > tissue need to be in the fluid for the 'fixation' to occur? > > Many thanks > > Paul Atkinson > > > DISCLAIMER: This e-mail is confidential and privileged. If you are not the > intended recipient please accept our apologies; please do not disclose, copy > or distribute information in this e-mail or take any action in reliance on > its contents: to do so is strictly prohibited and may be unlawful. Please > inform postmaster@rli.mbht.nhs.uk that this message has gone astray before > deleting it. Comments or opinions expressed in this email are those of > their respective contributors only. The views expressed do not represent the > views of the Trust, its management or employees. Morecambe Bay Hospitals NHS > Trust is not responsible and disclaims any and all liability for the content > of comments written within.Thank you for your co-operation. > > > _______________________________________________ > Histonet mailing list > Histonet@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 Sarah_Mack <@t> urmc.rochester.edu Wed Aug 24 09:52:26 2005 From: Sarah_Mack <@t> urmc.rochester.edu (Mack, Sarah) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] VEGF and VEGF-R in FFPE mouse Message-ID: I hope that this will help. Best of luck. VEGF Immuno Protocol Date Run: Step Minutes Temp Complete 1) Bake slides 60 60C 2) Deparaffinize & rehydrate to PBS RT 3) Methanolic 3% H2O2 30 RT 4) Rinse PBS X 3 15 RT 5) DAKO Proteinase K Digestion 6 RT 6) Rinse PBS X 3 5 RT 7) PAP around section RT 8) 10% Normal Goat Serum/PBS/BSA 30 RT 9) VEGF 1:200/PBS/BSA Overnight 4C 10) Rinse PBS X 3 15 RT 11) Bo Goat anti-rabbit IgG 1:500 30 RT 12) Make ABC RT 13) Rinse PBS X 3 15 RT 14) ABC complex 30 RT 15) Rinse PBS X 3 15 RT 16) 0.5% DAB 3-10 min RT 17) Rinse DH2O 3 RT 18) Counterstain Hematoxylin 1 RT 19) Dehydrate to Xylene RT 20) Cover with Permount RT Reagent Lot Numbers: Normal Goat Serum : Vector S-1000 : Lot # VEGF Antibody : Santa Cruz SC-507 : Lot # Bo Goat anti-rabbit : Vector BA-1000 : Lot # ABC Kit : Vector PK:6100 : Exp. DAB Kit : Vector SK:4100 : Exp. Notes: 1)All antibodies made with PBS &1% BSA & 0.1% Tween 2)Negative control Rabbit IgG 1:200 3) DAKO Digestion:1 Drop stock/ 2ml .05M Tris HCl pH7.5 > ---------- > From: Trajkovic, Dusko > Sent: Wednesday, August 24, 2005 9:23 AM > To: Mack, Sarah > Subject: RE: [Histonet] VEGF and VEGF-R in FFPE mouse > > Good morning Sarah, > I think many of us would love to see a complete protocol procedure for the > VEGF antibody. In the past, I have tried many different procedures and > none > of them resulted in any meaningful success. > Thank you in advance and have a wonderful day. > Dusko Trajkovic > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mack, > Sarah > Sent: Wednesday, August 24, 2005 5:43 AM > To: histonet@lists.utsouthwestern.edu; > 'wasielewski.reinhard.von@mh-hannover.de' > Subject: RE: [Histonet] VEGF and VEGF-R in FFPE mouse > > We use Santa Cruz VEGF antibody, catalog #SC-507. > > > Sarah A Mack > University of Rochester > Cardiovascular Research Institute (CRI) > 575 Elmwood Avenue, MRBX 2-11001-A > Rochester, NY 14572 > 585-273-1914 > > > > ---------- > > From: wasielewski.reinhard.von@mh-hannover.de > > Sent: Wednesday, August 24, 2005 8:22 AM > > To: histonet@lists.utsouthwestern.edu > > Subject: [Histonet] VEGF and VEGF-R in FFPE mouse > > > > Dear Histonetters, > > may I ask your help to find out the best antibody and staining procedure > > for formalin- > > fixed, paraffin embedded mouse tissue for the markers VEGF and VEGFR ? > > > > Many thanks in advance - best regards > > > > Reinhard > > > > > > > > PD Dr. med. Reinhard von Wasielewski > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > LEGAL NOTICE > Unless expressly stated otherwise, this message is confidential and may be > privileged. It is intended for the addressee(s) only. Access to this > E-mail by anyone else is unauthorized. If you are not an addressee, any > disclosure or copying of the contents of this E-mail or any action taken > (or not taken) in reliance on it is unauthorized and may be unlawful. If > you are not an addressee, please inform the sender immediately. > > From kwalker <@t> ils-inc.com Wed Aug 24 10:07:04 2005 From: kwalker <@t> ils-inc.com (Kimwa Walker) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] antibody needed Message-ID: <000201c5a8bd$7d531790$1802a8c0@ilsinc.com> Hi, The laboratory I'm working in is currently trying to complete IHC assays on tissues for an important study of uterine fibroid tissues. The antibody we've been utilizing is the Rabbit Polyclonal version of Ki-67 purchased from Vector Laboratories (catalog# VP-K451, lot# 301107), in conjunction with their Alkaline Phosphatase Detection Kit. We've recently exhausted our supply of this lot of antibody and have tried the assay with Vector's newest lot # for this antibody, but haven't had a successful stain at all. So, we'd like to complete the remainder of the slides we need to stain with the original lot that we've been using. Unfortunately, Vector Labs no longer has any supply of the #301107 rabbit polyclonal batch left either, so I wanted to see if anyone out there might have this particular batch of Ki-67 in their stock (it has to be the specific clone and lot# as mentioned above) that our lab might arrange to borrow or purchase from you. If you don't have the antibody yourself, but know where else it might be found, I'd like to hear about that too. Any assistance anyone can provide would be hugely appreciated. Thanks much! Kimwa L. Walker, B.S., HTL(ASCP) Histology Technician III ILS P.O. Box 13501 RTP, NC 27709 (919) 281-1110, ext 731 (919) 281-1118 (fax) kwalker@ils-inc.com www.ils-inc.com From ryaskovich <@t> dir.nidcr.nih.gov Wed Aug 24 10:12:32 2005 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR)) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Headache X pregnancy Message-ID: It is a major issue. I just went to another room and cut for 9 months. Don't go near it! Ruth Yaskovich National Institutes of Health National Institute of Dental and Crainiofacial Research Pain and Neurosensory Mechanism Branch > ---------- > From: Mike Safron > Reply To: Mike Safron > Sent: Monday, August 22, 2005 2:32 PM > To: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Headache X pregnancy > > If you are pregnant, or employ a tech who is, please take a look at the > results of this toxicology study. I think that xylene use by those > pregnant > is a major issue in histology. > > Pregnancy Outcome Following Gestational Exposure to Organic Solvents: A > Prospective Controlled Study > JAMA, Mar 1999; 281: 1106 - 1109. > > Michael Safron A.S.,HT(ASCP) > Manager, Histology > WIL Research Laboratories LLC > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From RStaskiewicz <@t> agr.state.il.us Wed Aug 24 10:50:22 2005 From: RStaskiewicz <@t> agr.state.il.us (RStaskiewicz@agr.state.il.us) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Stain for fungi in cytological smear Message-ID: Good morning all! I have a request to stain a cytological smear of a nasal wash for cryptococcus. I have never stained smears for fungi, just tissue. Does anyone have any experience or suggestions? Rae Ann Staskiewicz HT(ASCP) Section Head-Histology Galesburg Animal Disease Lab 2100 S. Lake Storey Rd Galesburg, IL 61401 (309) 344-2451 rstaskiewicz@agr.state.il.us From ree3 <@t> leicester.ac.uk Wed Aug 24 10:53:38 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] RE: FERROPORTIN ANTIBODY Message-ID: I am after an antibody to ferroportin that works on rodent paraffin processed tissues..... Thanks Richard Edwards MRC TOXICOLOGY UNIT..U.K..... From Stephen.J.Scholz <@t> osfhealthcare.org Wed Aug 24 11:10:00 2005 From: Stephen.J.Scholz <@t> osfhealthcare.org (Scholz, Stephen J.) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Stain for fungi in cytological smear Message-ID: <7F1312711CA7474A89B3DF8BA0BA54D01333AE@pmc-rfd-mx01.intranet.osfnet.org> We stain a lot of cytology slides for fungus with the GMS method. The bad news is they are a bit temperamental when it comes to cells staying on the slide. I strongly suggest that you use charged or subbed slides (we use fisher's tissue path superfrost plus gold slides only for this application--pricy little buggers) and fix the smears for 10 minutes in 95%ETOH. This is the process that has given us the most success although there still is an occasion when the cells wash off during the stain. Good luck. Stephen Scholz HT(ASCP) St. Anthony Medical Center Rockford IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of RStaskiewicz@agr.state.il.us Sent: Wednesday, August 24, 2005 10:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Stain for fungi in cytological smear Good morning all! I have a request to stain a cytological smear of a nasal wash for cryptococcus. I have never stained smears for fungi, just tissue. Does anyone have any experience or suggestions? Rae Ann Staskiewicz HT(ASCP) Section Head-Histology Galesburg Animal Disease Lab 2100 S. Lake Storey Rd Galesburg, IL 61401 (309) 344-2451 rstaskiewicz@agr.state.il.us _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tahseen <@t> brain.net.pk Wed Aug 24 04:05:06 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Stain for fungi in cytological smear References: Message-ID: <000f01c5a88a$ed975bc0$972bfea9@m7c0y4> We are doing GMS Stain for fungi in cytological smear. Muhammad Tahseen Histology Supervisor SKMCH & RC Lahore Pakistan. ----- Original Message ----- From: To: Sent: Thursday, August 25, 2005 3:50 AM Subject: [Histonet] Stain for fungi in cytological smear > Good morning all! I have a request to stain a cytological smear of a nasal > wash for cryptococcus. I have never stained smears for fungi, just tissue. > Does anyone have any experience or suggestions? > > > Rae Ann Staskiewicz HT(ASCP) > Section Head-Histology > Galesburg Animal Disease Lab > 2100 S. Lake Storey Rd > Galesburg, IL 61401 > (309) 344-2451 > rstaskiewicz@agr.state.il.us > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Aug 24 11:14:39 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] More on Re: Michel's medium for skin biopsies In-Reply-To: <082420051440.14304.430C86E00003FD4A000037E02200762194CECE0 30E9D0C9A03@comcast.net> References: <082420051440.14304.430C86E00003FD4A000037E02200762194CECE030E9D0C9A03@comcast.net> Message-ID: <6.0.0.22.1.20050824093445.01b553b0@gemini.msu.montana.edu> If I remember correctly, the Zeus tissue "fixative" IS Michels transport medium and merely made, packaged and marketed under the Zeus "fixative" name by Zeus Scientific although we purchased it through a vendor, Wampole (?) and in my estimation, calling this reagent Zeus fixative has caused confusion for time immemorial. Go to this website and read up on it www.zeusscientific.com . All information is complete on use,, etc with extensive references. WE considered making up Michels in house, but after long discussions with Jules Elias and reading the following publications: 1. J Elias. New method for shipment of renal biopsies. J Histotechnology (1(1):15-16, 1977 Elias indicated via private conversation it acted via protein denaturation in Michels or "fixative" solution which was then reversed by the rinse buffer before snap freezing, cryotomy and immunofluorescent staining. 2. Michel B et al,. Preservation of tissue fixed immunoglobulins in skin biopsies of patients with lupus erythematosis and bullous dieases - preliminary report (original publication). J Investigative Dermatology, 59(6):449-452, 1973 We decided to purchase it from the vendor under the "Zeus" fixative label along with the rinse buffer. We later changed supplier to Poly Scientific due to larger quantity for less expense and repackaged it ourselves to make up renal biopsy shipping kits for our customers. We had to have biopsies within 72 hours. At 08:40 AM 8/24/2005, you wrote: >Hi Paul, > >Michel's is a transport media not a fixative as Robyn indicated. It is >similar to Zeus in some components. It really depends on what you need a >fixative or something to move the tissue and preserve the antigenic sites >during that process. The transport media are generally only good for >holding 24 to 48 hours with most recommending 24 hours or less for EM. > >Good Luck, >Pam Marcum > >-------------- Original message -------------- > > > Paul, > > I believe Michels is a transport media, not a fixative media. You don't > want > > the tissue fixed, if you are doing antibody testing on it, in the fresh > tissue > > sense. I use Zeus Tissue Transport Media, my doctor seems to like it > better. > > > > Robyn > > OHSU > > > > >>> "Atkinson Paul B" 8/24/2005 2:18 AM >>> > > > > We are considering using Michels's transport medium for skin biopsies > in our > > laboratory. Have other laboratories found its use worthwhile? The original > > 1973 paper only mentions the effect on immunoglobulin staining, is it also > > effective in preserving complement antigenicity? Also how long does the > > tissue need to be in the fluid for the 'fixation' to occur? > > > > Many thanks > > > > Paul Atkinson > > > > > > DISCLAIMER: This e-mail is confidential and privileged. If you are not the > > intended recipient please accept our apologies; please do not disclose, > copy > > or distribute information in this e-mail or take any action in reliance on > > its contents: to do so is strictly prohibited and may be unlawful. Please > > inform postmaster@rli.mbht.nhs.uk that this message has gone astray before > > deleting it. Comments or opinions expressed in this email are those of > > their respective contributors only. The views expressed do not > represent the > > views of the Trust, its management or employees. Morecambe Bay > Hospitals NHS > > Trust is not responsible and disclaims any and all liability for the > content > > of comments written within.Thank you for your co-operation. > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From M.Prideaux <@t> sheffield.ac.uk Wed Aug 24 11:34:44 2005 From: M.Prideaux <@t> sheffield.ac.uk (Matt Prideaux) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Stain for fungi in cytological smear In-Reply-To: References: Message-ID: <1124901283.430ca1a401829@webmail.shef.ac.uk> The lab I used to work at did Grocott's staining for fungi on cytology smears regularly, without the cells washing off. I think the slides were APES treated. Hope that helps. -- Matt Prideaux Academic Unit of Bone Biology Division of Clinical Sciences (South) D Floor (DU20) Medical School Henry Wellcome Laboratories for Medical Research University of Sheffield Beech Hill Road Sheffield S10 2RX Quoting RStaskiewicz@agr.state.il.us: > Good morning all! I have a request to stain a cytological smear of a nasal > wash for cryptococcus. I have never stained smears for fungi, just tissue. > Does anyone have any experience or suggestions? > > > Rae Ann Staskiewicz HT(ASCP) > Section Head-Histology > Galesburg Animal Disease Lab > 2100 S. Lake Storey Rd > Galesburg, IL 61401 > (309) 344-2451 > rstaskiewicz@agr.state.il.us > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From TJJ <@t> Stowers-Institute.org Wed Aug 24 12:11:09 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Re: stain for fungi in cytological smear Message-ID: Rae, if you are staining for cryptococcus, Mucicarmine stains that quite well. I offer it as an alternative to a silver technique. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From Elizabeth.Martin2 <@t> HCAhealthcare.com Wed Aug 24 12:22:24 2005 From: Elizabeth.Martin2 <@t> HCAhealthcare.com (Martin Elizabeth - Arlington) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Re: Stain for fungi in cytological smear (Matt Prideaux) Message-ID: You can use either air-dried or alcohol fixed smears. Both work fine. If you are looking for cryptococcus and using a mucicarmine stain, then the alcohol looks better. GMS and PAS, I also prefer alcohol fixed, but the air-dried also works if that is all you have. Elizabeth Martin, M.D. Northern Virginia Community Hospital Arlington, VA Today's Topics: 1. Re: Stain for fungi in cytological smear (Matt Prideaux) ---------------------------------------------------------------------- Message: 1 Date: Wed, 24 Aug 2005 17:34:44 +0100 From: Matt Prideaux Subject: Re: [Histonet] Stain for fungi in cytological smear To: RStaskiewicz@agr.state.il.us, Histonet Message-ID: <1124901283.430ca1a401829@webmail.shef.ac.uk> Content-Type: text/plain; charset=ISO-8859-1 The lab I used to work at did Grocott's staining for fungi on cytology smears regularly, without the cells washing off. I think the slides were APES treated. Hope that helps. -- Matt Prideaux Academic Unit of Bone Biology Division of Clinical Sciences (South) D Floor (DU20) Medical School Henry Wellcome Laboratories for Medical Research University of Sheffield Beech Hill Road Sheffield S10 2RX Quoting RStaskiewicz@agr.state.il.us: > Good morning all! I have a request to stain a cytological smear of a nasal > wash for cryptococcus. I have never stained smears for fungi, just tissue. > Does anyone have any experience or suggestions? > > > Rae Ann Staskiewicz HT(ASCP) > Section Head-Histology > Galesburg Animal Disease Lab > 2100 S. Lake Storey Rd > Galesburg, IL 61401 > (309) 344-2451 > rstaskiewicz@agr.state.il.us > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 21, Issue 31 **************************************** From mfrei <@t> sial.com Wed Aug 24 13:22:13 2005 From: mfrei <@t> sial.com (Mark Frei) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] RE: Histonet Digest, Vol 21, Issue 29 In-Reply-To: <457381D92B01BD44B21CF37CC02EBDFD0292771D@vhacinexc2.v10.med.va.gov> Message-ID: The Sigma-Aldrich Hematology and Histology website, sigmaaldrich.com/handh, has a section for instrument applications for people to use or review. It does require a sign-in process in case we need to notify users of any changes or updates. Mark Frei MT(ASCP) Sigma-Aldrich Corporation 3050 Spruce Street St. Louis, MO 63103 (314) 286-8080 Julie.Sanders@med.va.gov Sent by: histonet-bounces@lists.utsouthwestern.edu 08/24/2005 07:15 AM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] RE: Histonet Digest, Vol 21, Issue 29 This question is for folks who use the Leica autostainer. I'd like to see various H&E protocols for this stainer and see what everyone is using. I appreciate all responses. Thanks, Julie Sanders, BA, HT(ASCP) Supervisor, Anatomic Pathology VAMC, Cincinnati, Ohio _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lowman034 <@t> yahoo.com Wed Aug 24 16:02:19 2005 From: lowman034 <@t> yahoo.com (Dave Low) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Autopsy Worksheet In-Reply-To: <4D95C24EC0E4C84787A6919F1165B25E24B1B6@btmhems01.BTHOSP.INT> Message-ID: <20050824210219.5556.qmail@web32007.mail.mud.yahoo.com> Sue, Check this link out from AFIP! Hopefully this works! http://www.afip.org/Departments/oafme/diagrams.html Dave --- "O'Brien, Sue" wrote: > Does anyone have an autopsy worksheet that they are > willing to share > with me? (I am looking for one that would be a > fill-in the blanks type > for organs, weights etc.). Anything you would have > as a start point > would be appreciated! Thank-you, > > Sue O'Brien, Histology Supervisor > > Burdette Tomlin Memorial Hospital > > E-mail: sobrien@bthosp.com > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________ Do you Yahoo!? Yahoo! Mail - You care about security. So do we. http://promotions.yahoo.com/new_mail From llewllew <@t> shaw.ca Tue Aug 23 23:20:46 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Using chromic acid instead of periodic acid with Schiffs - for fungus staining ONLY! Message-ID: <000601c5a863$33bc1500$7e034246@yourlk4rlmsu> Apparently I screwed up. The concentration of chromic acid should be 4% for 1 hour (or 10% for 10 minutes - a much later modification). Sorry. Bryan Llewellyn ----- Original Message ----- From: "Bryan Llewellyn" To: "Histonet" Sent: Tuesday, August 23, 2005 4:29 PM Subject: Re: [Histonet] Using chromic acid instead of periodic acid with Schiffs - for fungus staining ONLY! >I think the person wanted the details for Bauer's stain, or CAS (chromic >acid Schiff). It was originally published for glycogen, I believe, but PAS >is better for that. The method is simple. > > Oxidise with 1% chromium trioxide in water for 1 hour (or 5% for 10 > minutes). > Bleach with 5% metabisulphite and wash. > Schiff's reagent 20 minutes or so. > Finish off as you would a PAS. > > Bryan Llewellyn > > > > ----- Original Message ----- > From: "Gayle Callis" > To: > Sent: Tuesday, August 23, 2005 3:06 PM > Subject: [Histonet] Using chromic acid instead of periodic acid with > Schiffs - for fungus staining ONLY! > > >> Histonetters, >> >> I hope I didn't confuse people with my answer about PAS-F (what the F >> meant escapes me??? for fungus, fluorescence?) where the person was >> asking about using chromic acid instead of periodic acid i.e in PAS. >> Theoretically, one could not call this PAS - maybe chromic acid-Schiff or >> CAS?? The key word was the mention of Gridley staining, a method for >> fungus staining, and I assumed this person wanted to use a chromic acid >> oxidizer followed by Schiffs reagent for that purpose i.e fungus stain. >> >> Whatever you do, do NOT use chromic acid for a standard PAS stain if >> staining for mucosubstances, glycogen, basement membranes or other >> components that are PAS positive. Chromic acid is a much stronger >> oxidizer than periodic acid, and will over-oxidize these components to >> the point of them NOT staining after Schiffs application - not a good >> idea. However chromic acid with Schiffs does work for fungus staining. >> Freida Carson et al wrote a publication on false negative fungus >> staining by using Periodic acid -Schiffs reagent, and mentioned using >> chromic acid in place of periodic acid. This can be found in J of >> Histotechnology, and an excellent bit of information. >> >> Chromic acid oxidation with Schiffs reagents is reserved for those who do >> not want use periodic acid as the oxidizing agent for fungus staining. >> >> Gayle Callis >> Research Histopathology Supervisor >> Veterinary Molecular Biology >> Montana State University - Bozeman >> PO Box 173610 >> Bozeman MT 59717-3610 >> 406 994-6367 >> 406 994-4303 (FAX) >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > From cbass <@t> bidmc.harvard.edu Wed Aug 24 17:04:32 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] HCV core-E1-E2 antibody Message-ID: Hi, Does anyone have suggestions for an antibody against HCV core-E1-E2, or any of the single genes? I would like something that would work with western or IHC. However, as we will probably do more IHC I would like one that works specifically with it. I have found a few antibodies that "should" work with tissue, but the company has only tested it with westerns. The only problem is that we need a non- mouse antibody. Any other species will be fine. Thanks, Caroline Bass From AnthonyH <@t> chw.edu.au Wed Aug 24 17:59:12 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Stain for fungi in cytological smear Message-ID: The smears can either be air-dried or ethanol fixed. I have found better cellular retention with air-drying. After fixation (see above) do a PAS, or a mucicarmine or the Alcian Blue-Argentaffin reaction for the cryptococcus. For other fungi, a PAS or Chromic acid-Methenamine Silver or a Sulphated Toluidine Blue (good for pneumocystis)can be done. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of RStaskiewicz@agr.state.il.us Sent: Thursday, 25 August 2005 1:50 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Stain for fungi in cytological smear Good morning all! I have a request to stain a cytological smear of a nasal wash for cryptococcus. I have never stained smears for fungi, just tissue. Does anyone have any experience or suggestions? Rae Ann Staskiewicz HT(ASCP) Section Head-Histology Galesburg Animal Disease Lab 2100 S. Lake Storey Rd Galesburg, IL 61401 (309) 344-2451 rstaskiewicz@agr.state.il.us _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From gcallis <@t> montana.edu Wed Aug 24 18:12:27 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] NSH VIR committee table volunteers needed Message-ID: <6.0.0.22.1.20050824170835.01b5af98@gemini.msu.montana.edu> Are there any Veterinary Industry and Research Committee members attending the Florida NSH convention meeting who could help man the VIR committee table on Sat, Sunday, Monday or Tuesday. This is done during breaks and noon hour. If so, please contact the VIR chair Diane Sterchi at d.sterchi@lilly.com so she can schedule a time. It would certainly help out. Thanks Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From Linresearch <@t> aol.com Wed Aug 24 18:49:49 2005 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] GLP Guidelines Message-ID: <194.45a48d1d.303e619d@aol.com> Hello, I am trying to find GLP guidelines for computer data and images. If anyone knows of a site or has the current guidelines, could send me a copy or direct me to the site? Thanks, Lin From frintelen <@t> yahoo.com Thu Aug 25 02:53:29 2005 From: frintelen <@t> yahoo.com (Felix Rintelen) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Differentiation between Eosinophil and Neutrophil by Sirius Red staining? Message-ID: <20050825075329.55234.qmail@web35302.mail.mud.yahoo.com> Hi everyone, I'm trying to decide by histological staining whether an infiltrate in mouse tissue is dominated by either neutrophils or eosinophils. I performed the Llewllyn's sirius red F3B staining (see web page http://stainsfile.info/StainsFile/stain/amyloid/siriusllew.htm, Sirius Red F3B = Direct Red 80 was from Sigma Cat N? 365548) which according to the protocol stains Eosinophils and Paneth cells. This staining was also recommended in the Histonet and indeed the staining worked very nicely. Now, my question is just, if anybody knows if neutrophils are not stained by this procedure or not. In other words, if I can be sure that when a cell is stained red, it will for sure be an eosinophil and not a neutrophil. Thank you very much in advance! Best regards, Felix Felix Rintelen Serono Pharmaceutical Research Institute Geneva, Switzerland --------------------------------- Do you Yahoo!? Read only the mail you want - Yahoo! Mail SpamGuard. From RitaR <@t> lexhealth.org Thu Aug 25 05:51:10 2005 From: RitaR <@t> lexhealth.org (Rita Riddle) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] CAS or PAS-F Message-ID: Thanks so much for all the great response to my question regarding chromic acid. I tried one and the Dr. just loved it. Great work Histo. Rita Riddle HT(ASCP) Lead Histology Tech Lexington Medical Center West Columbia, SC 29169 Telephone 803-791-2881 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From TJJ <@t> Stowers-Institute.org Thu Aug 25 08:34:56 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Leica RM2255 holder for round specimens Message-ID: I just bought the accessory for our RM2255 microtome so we could use it for sectioning our PMMA embedded samples. The inserts that came with it are 6, 15, and 25 mm in diameter. It appears that unless the diameter of the block is almost exactly this measurement, the holder will not work. Is this truly the case or is there some adjustment we aren't aware of? Unfortunately as well, the vials we used for embedding are smaller than 25 mm but larger than 15. We plan to either mount them onto a block holder with adhesive and try to cut them that way, or use a dremel or other power tool to reduce the diameter TO 15 mm. Are there other things we can do that I'm just not seeing? Thanks! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From jkiernan <@t> uwo.ca Thu Aug 25 09:03:35 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Differentiation between Eosinophil and Neutrophil by SiriusRed staining? References: <20050825075329.55234.qmail@web35302.mail.mud.yahoo.com> Message-ID: <430DCFB7.44F7056F@uwo.ca> Bryan Llewellyn's method uses sirius red F3B from an alkaline solution, in contrast to the methods for collagen (Puchtler, Junqueira etc) in which the same dye is dissolved in saturated aqueous picric acid. An alkaline solution of an anionic dye will be attracted only to strongly basic proteins - those rich in arginine. (The guanidino side-chain of arginine is a cation even at high pH.) This limits the staining to eosinophil granules, Paneth cell granules and the tails of spermatozoa. Amyloid is also stained - the principal application of the method - but that's for different reasons. Alkaline solutions of some anionic dyes can stain elastic fibres and laminae (also for different reasons). Stained eosinophils are unlikely to be confuesed with any of the other things that can be coloured by an alkaline solution of an acid dye. The dye doesn't have to be sirius red F3B. Eosin Y will do the job; that's how the cells got their name. John Kiernan Anatomy, UWO London, Canada. ______________________________________________ Felix Rintelen wrote: > > Hi everyone, > > > > I'm trying to decide by histological staining whether an infiltrate in mouse tissue is dominated by either neutrophils or eosinophils. I performed the Llewllyn's sirius red F3B staining (see web page http://stainsfile.info/StainsFile/stain/amyloid/siriusllew.htm, Sirius Red F3B = Direct Red 80 was from Sigma Cat N? 365548) which according to the protocol stains Eosinophils and Paneth cells. This staining was also recommended in the Histonet and indeed the staining worked very nicely. > > Now, my question is just, if anybody knows if neutrophils are not stained by this procedure or not. In other words, if I can be sure that when a cell is stained red, it will for sure be an eosinophil and not a neutrophil. > > > > Thank you very much in advance! > > > > Best regards, > > > > Felix > > > > > > Felix Rintelen > > Serono Pharmaceutical Research Institute > > Geneva, Switzerland > > > --------------------------------- > 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 Don.Birgerson <@t> leica-microsystems.com Thu Aug 25 09:07:44 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Leica RM2255 holder for round specimens In-Reply-To: <20050825133751.7B227D1F3FD9@NA-Barracuda.leica-microsystems.com> Message-ID: Hi Teri, I suggest you use the "Standard Specimen Clamp with the "V" notch insert. Plastic embedded specimens are hard enough to clamp directly without crushing. (14050237998 and 14050238000 ). If you have further questions, please call. Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Johnson, Teri" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/25/2005 08:34 AM To "Histonet" cc Subject [Histonet] Leica RM2255 holder for round specimens I just bought the accessory for our RM2255 microtome so we could use it for sectioning our PMMA embedded samples. The inserts that came with it are 6, 15, and 25 mm in diameter. It appears that unless the diameter of the block is almost exactly this measurement, the holder will not work. Is this truly the case or is there some adjustment we aren't aware of? Unfortunately as well, the vials we used for embedding are smaller than 25 mm but larger than 15. We plan to either mount them onto a block holder with adhesive and try to cut them that way, or use a dremel or other power tool to reduce the diameter TO 15 mm. Are there other things we can do that I'm just not seeing? Thanks! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com From boycea <@t> mail.nih.gov Thu Aug 25 09:30:38 2005 From: boycea <@t> mail.nih.gov (Boyce, Amanda (NIH/NIAMS)) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] muscle digest for sekeltal prep Message-ID: <0E3E7E8F6E23DF4C8127A063568356B510B98792@nihexchange12.nih.gov> We are processing animals for skeletal preps that have been in 10% PFA for quite some time (many months). The first steps involved skinning and eviscerating the embryos, followed by alcian blue staining. We then put the embryos in a trypsin digetion solution, which normally chews up the majority of the muscle. The rest of the muscle will be dissolved after the skeletons are stained with alizarin red and put through a series of KOH washes. The problem is that because these animals are overfixed, the trypsin digest is not sufficient. Does anyone have any suggestions for a more vigorous digestion? Thanks for your help! Amanda -------------------------- Amanda Taylor Boyce, Ph.D. Postdoctoral IRTA Fellow Cartilage Biology and Orthopaedics Branch National Institute of Arthritis and Musculoskeletal and Skin Diseases National Institutes of Health Bldg. 13, Rm. 3W17 Bethesda, MD 20892-5755 Phone: 301-451-6860 Fax: 301-480-4315 Email: boycea@mail.nih.gov From juan.gutierrez <@t> christushealth.org Thu Aug 25 09:50:11 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Hospital fetus disposition survey Message-ID: I'll try to make this short. We are a Catholic not for profit hospital. Parents have the option to bury on their own as long as they go thru a funeral home regardless of gestational Age. The hospital also offers a free mass burial once a month for the <20 week group. For the 20> week they are offered a private burial at a greatly reduced price or free if they can't afford it. In the case when the parents do not care about the remains, they are buried as stated above, with the hospital incurring the cost. In Texas, state law only requires that a funeral home handles the transaction. They don't care what the final disposition is. 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 chiggerson@memhosp.com Sent: Tuesday, August 23, 2005 11:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Hospital fetus disposition survey Histonetters, We are evaluating our current practices for fetus disposal at our facility and would like to know how both Catholic and non-Catholic hospitals handle fetus disposition at their facilities. I am interested in practices for both fetal loss greater than 20 weeks gestation AND less than 20 weeks gestation (products of conception). Please include religious affiliation, if any, of your institution. 1. Are parents given the option to have the hospital handle the disposition for the remains at < 20 wks, >20 weeks, or both? 2. If parents opt for hospital disposition, is it handled on-site, or contracted by the hospital off-site (with a funeral home)? If off-site, is the patient charged for disposition? 3. Method of disposition? Cremation? Burial? Other? 4. Are remains collected/disposed of separately, or co-mingled with other tissues? 5. Is disposal method dictated by State Law? Please include state. Any additional information you can give me about your practices would be helpful. Thank you in advance for your help. Cindy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Aug 25 10:33:13 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] NSH IHCRG committee table volunteers needed In-Reply-To: <6.0.0.22.1.20050824170835.01b5af98@gemini.msu.montana.edu> Message-ID: <200508251533.j7PFXCnQ003569@chip.viawest.net> I would like to ditto Gayles email asking for help with VIR committee table at the NSH S/C for the IHC Resource Group. If there are any members of the IHCRG attending the NSH S/C in Florida willing to help man our tables, please contact Patsy Ruegg at pruegg@ihctech.net If you are not yet a member of the IHC Resource Group but want to be, you can sign up online at www.ihcrg.org Thanks, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Wednesday, August 24, 2005 4:12 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] NSH VIR committee table volunteers needed Are there any Veterinary Industry and Research Committee members attending the Florida NSH convention meeting who could help man the VIR committee table on Sat, Sunday, Monday or Tuesday. This is done during breaks and noon hour. If so, please contact the VIR chair Diane Sterchi at d.sterchi@lilly.com so she can schedule a time. It would certainly help out. Thanks Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MSafron <@t> wilresearch.com Thu Aug 25 10:57:00 2005 From: MSafron <@t> wilresearch.com (Mike Safron) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Leica RM2255 holder for round specimens Message-ID: Teri, If you decide to mount them onto block holders you will need to order them from Leica. Many of the other block holders will not fit the chucks provided (I know they look the same but trust me.) The item number for the block holders is 702218310. You can check their website to make sure that is what you are thinking of. If you have any questions feel free to call. Michael Safron A.S.,HT(ASCP) Manager, Histology WIL Research Laboratories LLC 419-289-8700 ext: 3040 Ashland, Ohio 44805 From dmarsha3 <@t> utmem.edu Thu Aug 25 11:15:47 2005 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] polyvinylpyrollidone-edta recipe request Message-ID: <000f01c5a990$40910dc0$f623c084@DanaM> Hi everyone, If someone could take pity on me and send their recipe for a PVP-EDTA decalcification/fixation solution I would really appreciate it. I have to take tissues really soon, need to put bones in PVP solution and cannot find a recipe. I am an amateur and appreciate your help:-) Thanks, Dana From gcallis <@t> montana.edu Thu Aug 25 11:16:38 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Leica RM2255 holder for round specimens In-Reply-To: <200508251340.j7PDejHe026813@mail4.msu.montana.edu> References: <200508251340.j7PDejHe026813@mail4.msu.montana.edu> Message-ID: <6.0.0.22.1.20050825100828.01b82e08@gemini.msu.montana.edu> We ground down our blocks to fit in whatever holder we had OR superglue the block to a correct diameter sized but blank PMMA block - we just made a bunch of those to keep around. The ends of the bone block and blank block being interfaced should be flat surfaces but rough to ensure the superglue really holds at the interface, like gluing a slab section to a plastic slide. You don't want blocks popping off during trim or microtoming. In fact, we would grind the block face down to the bone before trimming to save the tungsten carbide edge and speed up trimming this plastic. PMMA will need rock solid adhesive holding and you do not sacrifice your holder to some messy glue. At 07:34 AM 8/25/2005, you wrote: >I just bought the accessory for our RM2255 microtome so we could use it >for sectioning our PMMA embedded samples. The inserts that came with it >are 6, 15, and 25 mm in diameter. It appears that unless the diameter >of the block is almost exactly this measurement, the holder will not >work. Is this truly the case or is there some adjustment we aren't >aware of? Unfortunately as well, the vials we used for embedding are >smaller than 25 mm but larger than 15. We plan to either mount them >onto a block holder with adhesive and try to cut them that way, or use a >dremel or other power tool to reduce the diameter TO 15 mm. Are there >other things we can do that I'm just not seeing? > >Thanks! > >Teri Johnson >Managing Director Histology Facility >Stowers Institute for Medical Research >1000 E. 50th St. >Kansas City, MO 64133 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From liz <@t> premierlab.com Thu Aug 25 11:16:44 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] angiomyolipoma of kidney Message-ID: <001201c5a990$6267ed10$a7d48a80@AMY> Does anyone know of a source that I could get blocks from an angiomyolipoma of kidney. I'm also looking for a good source for normal tissue microarrays. Any suggestions would be helpful. Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 MSafron <@t> wilresearch.com Thu Aug 25 11:22:38 2005 From: MSafron <@t> wilresearch.com (Mike Safron) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] GLP Guidelines Message-ID: Here is the FDA website for part 11 issues. Make sure you have plenty of coffee......it is not too exciting, and it is also somewhat vague in parts. Good Luck. http://www.fda.gov/cder/guidance/5667fnl.htm Michael Safron A.S.,HT(ASCP) Manager, Histology WIL Research Laboratories LLC 419-289-8700 ext: 3040 Ashland, Ohio 44805 From joycefr <@t> frontiernet.net Thu Aug 25 11:49:39 2005 From: joycefr <@t> frontiernet.net (Joyce Friedland) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Microwave Tissue Processors Message-ID: <6cf02f167b6109bcaacf0454af7097e5@frontiernet.net> Hi All, My lab director is interested in purchasing a processor which will allow us to process batches of cases throughout the day, and serve as a backup to our two VIPs. I would greatly appreciate knowing what experiences you all have had with the various microwave processors available. I have particular interest in the Pathos from Milestone since it is automated, but I am open to all possibilities. Thanks, Joyce Friedland From jqb7 <@t> cdc.gov Thu Aug 25 11:47:45 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Microwave Tissue Processors Message-ID: Be sure and look into the Sakura Xpress as well. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Friedland Sent: Thursday, August 25, 2005 12:50 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave Tissue Processors Hi All, My lab director is interested in purchasing a processor which will allow us to process batches of cases throughout the day, and serve as a backup to our two VIPs. I would greatly appreciate knowing what experiences you all have had with the various microwave processors available. I have particular interest in the Pathos from Milestone since it is automated, but I am open to all possibilities. Thanks, Joyce Friedland _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DonnaWillis <@t> texashealth.org Thu Aug 25 12:05:03 2005 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Microwave Tissue Processors Message-ID: <4483B6EE0FA82A4EAED2B1E161E5E41147C9B4@ftwex02.txhealth.org> Joyce, The Pathos will allow you to do all that your are interested in doing on biopsy samples and on tissue cut up to 5mm thick. I have one in my lab. You can batch up to 210 cassettes. I have not worked with the Sakura unit but I know that the thickness of the samples on the unit are limited and batch volumes are smaller. Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Friedland Sent: Thursday, August 25, 2005 11:50 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave Tissue Processors Hi All, My lab director is interested in purchasing a processor which will allow us to process batches of cases throughout the day, and serve as a backup to our two VIPs. I would greatly appreciate knowing what experiences you all have had with the various microwave processors available. I have particular interest in the Pathos from Milestone since it is automated, but I am open to all possibilities. Thanks, Joyce Friedland _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From lrichey <@t> u.washington.edu Thu Aug 25 12:14:38 2005 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Microwave Tissue Processors In-Reply-To: <6cf02f167b6109bcaacf0454af7097e5@frontiernet.net> References: <6cf02f167b6109bcaacf0454af7097e5@frontiernet.net> Message-ID: <430DFC7E.70204@u.washington.edu> We are unsing the Sakura express microwave processor. We process and cut sm biopsies throughout the day. It works especially well for rush specimens. It can be loaded every 15 minutes. We are still processing derms and breast bxs on our VIPs. Like all new stains and procedures, it takes some time for the pathologists to get use to the different process. Joyce Friedland wrote: > Hi All, > My lab director is interested in purchasing a processor which will > allow us to process batches of cases throughout the day, and serve as > a backup to our two VIPs. > I would greatly appreciate knowing what experiences you all have had > with the various microwave processors available. I have particular > interest in the Pathos from Milestone since it is automated, but I am > open to all possibilities. > > Thanks, > Joyce Friedland > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Thu Aug 25 12:23:28 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] muscle digest for sekeltal prep Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175BE@lsexch.lsmaster.lifespan.org> Apart from the obvious step of using a higher concentration of enzyme, you could try a mixture of trypsin and pepsin. Also, formaldehyde fixation can be largely undone by extended washing. Try washing the tissue in running water for 24 hours or longer, before doing the digestion procedure. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Boyce, Amanda (NIH/NIAMS) > Sent: Thursday, August 25, 2005 7:30 AM > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] muscle digest for sekeltal prep > > We are processing animals for skeletal preps that have been in 10% PFA for > quite some time (many months). The first steps involved skinning and > eviscerating the embryos, followed by alcian blue staining. We then put > the > embryos in a trypsin digetion solution, which normally chews up the > majority > of the muscle. The rest of the muscle will be dissolved after the > skeletons > are stained with alizarin red and put through a series of KOH washes. The > problem is that because these animals are overfixed, the trypsin digest is > not sufficient. Does anyone have any suggestions for a more vigorous > digestion? Thanks for your help! Amanda > > -------------------------- > Amanda Taylor Boyce, Ph.D. > Postdoctoral IRTA Fellow > Cartilage Biology and Orthopaedics Branch > National Institute of Arthritis and Musculoskeletal and Skin Diseases > National Institutes of Health > Bldg. 13, Rm. 3W17 > Bethesda, MD 20892-5755 > Phone: 301-451-6860 > Fax: 301-480-4315 > Email: boycea@mail.nih.gov > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From liz <@t> premierlab.com Thu Aug 25 12:36:38 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] eosinophil major basic protein Message-ID: <000001c5a99b$8cfea1d0$a7d48a80@AMY> Hello everyone Is anyone aware of an antibody to eosinophil major basic protein that will work in paraformaldehyde or formalin fixed guinea pig tissue. Thanks Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 emry <@t> u.washington.edu Thu Aug 25 12:37:49 2005 From: emry <@t> u.washington.edu (Trisha Emry) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] paraffin Message-ID: I would like to know if there are any new paraffins that would be better for bone and larger specimens like pig snouts. Thanks, Trisha Seattle From emry <@t> u.washington.edu Thu Aug 25 12:41:57 2005 From: emry <@t> u.washington.edu (Trisha Emry) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] microtome adjustment Message-ID: I am having problems with the microtome skipping every other slice. If there a blade adjustment that can help, please, describe how I can do that before I send it off to repair. Thanks, Trisha Seattle From djohnson14 <@t> hotmail.com Thu Aug 25 13:54:16 2005 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Please help us name our new product Message-ID: Mercedes Medical is coming out with a new teflon coated microtome blade, but we need a name! We have 3 names up for consideration: 1. Advant-Edge 2. Cutting Edge 3. Platinum Blade. Let us know which you like, or come up with a new one and if we use it we’ll send you a great gift. Also, come see us at our island booth at NSH in Ft. Lauderdale booth #229. Enter to win a 26” Flat screen TV, and pick up your free Henry the Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be held at the Annual Symposium/Convention from September 10-14 in Ft Lauderdale. You can register on line at www.NSH.org. Visit us on line at www.MercedesMedical.com and get 10% off your first order. Or call at 800.331.2716. From gcallis <@t> montana.edu Thu Aug 25 13:58:25 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] paraffin In-Reply-To: References: Message-ID: <6.0.0.22.1.20050825125552.01b44ce8@gemini.msu.montana.edu> You did not say what you used, but we liked Tissue Prep 2 (Fisher) - it is harder that helps support bone better and worked well with midsaggital slabs from sheep tibia (big slabs mounted on 4 x 5 inch slides) , goat bones, human femoral heads, etc. At 11:37 AM 8/25/2005, you wrote: >I would like to know if there are any new paraffins that would be better for >bone and larger specimens like pig snouts. > >Thanks, >Trisha >Seattle > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From kbroomal <@t> NEMOURS.ORG Thu Aug 25 14:16:26 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Please help us name our new product Message-ID: <6E41111281623B4B8A9AB8F9A7EA34378243FB@wlmmsx02.nemours.org> Advant Edge is the name of nutritional products. http://www.easadvantedge.com/adv_products.asp Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Dave Johnson Sent: Thursday, August 25, 2005 2:54 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Please help us name our new product Mercedes Medical is coming out with a new teflon coated microtome blade, but we need a name! We have 3 names up for consideration: 1. Advant-Edge 2. Cutting Edge 3. Platinum Blade. Let us know which you like, or come up with a new one and if we use it we'll send you a great gift. Also, come see us at our island booth at NSH in Ft. Lauderdale booth #229. Enter to win a 26" Flat screen TV, and pick up your free Henry the Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be held at the Annual Symposium/Convention from September 10-14 in Ft Lauderdale. You can register on line at www.NSH.org. Visit us on line at www.MercedesMedical.com and get 10% off your first order. Or call at 800.331.2716. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Don.Birgerson <@t> leica-microsystems.com Thu Aug 25 14:20:08 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] microtome adjustment In-Reply-To: Message-ID: Hi Trisha, This is most commonly caused by blade holder and its clearance angle being to small. Give me a call and I'll go over some things you can try. Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Trisha Emry" Sent by: histonet-bounces@lists.utsouthwestern.edu 08/25/2005 12:41 PM To "histo" cc Subject [Histonet] microtome adjustment I am having problems with the microtome skipping every other slice. If there a blade adjustment that can help, please, describe how I can do that before I send it off to repair. Thanks, Trisha Seattle _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com From petepath <@t> yahoo.com Thu Aug 25 14:35:59 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Histonet] Please help us name our new product Message-ID: <20050825193559.30481.qmail@web30406.mail.mud.yahoo.com> How about " Nasty Cut" blades. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From Jackie.O'Connor <@t> abbott.com Thu Aug 25 14:39:32 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Histonet] Please help us name our new product Message-ID: How about "A Cut Above". "Stephen Peters M.D." Sent by: histonet-bounces@lists.utsouthwestern.edu 08/25/2005 02:35 PM To: Histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Histonet] Please help us name our new product How about " Nasty Cut" blades. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Aug 25 14:45:05 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Microwave Tissue Processors In-Reply-To: <430DFC7E.70204@u.washington.edu> Message-ID: <200508251945.j7PJj4nQ020632@chip.viawest.net> I am interested in feed back on IHC results when using MW processors for tissue. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lori Richey Sent: Thursday, August 25, 2005 10:15 AM To: Joyce Friedland Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Microwave Tissue Processors We are unsing the Sakura express microwave processor. We process and cut sm biopsies throughout the day. It works especially well for rush specimens. It can be loaded every 15 minutes. We are still processing derms and breast bxs on our VIPs. Like all new stains and procedures, it takes some time for the pathologists to get use to the different process. Joyce Friedland wrote: > Hi All, > My lab director is interested in purchasing a processor which will > allow us to process batches of cases throughout the day, and serve as > a backup to our two VIPs. > I would greatly appreciate knowing what experiences you all have had > with the various microwave processors available. I have particular > interest in the Pathos from Milestone since it is automated, but I am > open to all possibilities. > > Thanks, > Joyce Friedland > > > _______________________________________________ > Histonet mailing list > Histonet@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 funderwood <@t> mcohio.org Thu Aug 25 14:44:17 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Histonet] Please help us name our new product Message-ID: Maybe Bono and Edge. >>> "Jackie M O'Connor" 08/25/05 03:39PM >>> How about "A Cut Above". "Stephen Peters M.D." Sent by: histonet-bounces@lists.utsouthwestern.edu 08/25/2005 02:35 PM To: Histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Histonet] Please help us name our new product How about " Nasty Cut" blades. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.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 Janet.Bonner <@t> FLHOSP.ORG Thu Aug 25 14:47:23 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Histonet] Please help us name our new product Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB437E@fh2k093.fhmis.net> I believe this is the name of a Hair cutting-Styling establishment. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Stephen Peters M.D. Cc: Histonet@lists.utsouthwestern.edu Sent: 8/25/2005 3:39 PM Subject: Re: [Histonet] Histonet] Please help us name our new product How about "A Cut Above". "Stephen Peters M.D." Sent by: histonet-bounces@lists.utsouthwestern.edu 08/25/2005 02:35 PM To: Histonet@lists.utsouthwestern.edu cc: Subject: [Histonet] Histonet] Please help us name our new product How about " Nasty Cut" blades. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.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 pkarlisch <@t> psu.edu Thu Aug 25 15:07:50 2005 From: pkarlisch <@t> psu.edu (Patricia Karlisch) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts Message-ID: Hello all, Can anyone reference a description for 'levels' vs 'recuts'. We use the terms to mean specific procedures in cutting. Thus, RECUTS are sections directly off the top of a previously cut block. Levels are 'recuts' but at different distances between sections. Generally, when we cut levels we toss the unwanted sections out. In some cases the unstained are kept. Does anyone in histology consider these to be 'routine' terms and is this defined in any textbook? I appreciate your input on this rather basic question. Pat Karlisch Supervisor, Histology, Pathology and Laboratory Medicine Penn State Milton S. Hershey Medical Center Mail Code H179 Hershey, PA 17033 Phone (717) 531-6072 Fax: (717) 531- 7741 email: pkarlisch@psu.edu *****E-Mail Confidentiality Notice***** This message (including any attachments) contains information intended for a specific individual(s) and purpose that may be privileged, confidential or otherwise protected from disclosure pursuant to applicable law. Any inappropriate use, distribution or copying of the message is strictly prohibited and may subject you to criminal or civil penalty. If you have received this transmission in error, please reply to the sender indicating this error and delete the transmission from your system immediately. From TCostello <@t> gtxinc.com Thu Aug 25 15:16:56 2005 From: TCostello <@t> gtxinc.com (Terry Costello) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Please help us name our new product Message-ID: <9E8CB17DF8638A46832960D5E3DCAACDC83330@gtxmail1.gtx.gtxinc.com> "MERCEDES BLADZ" Terrence Costello GTx, Inc. Three North Dunlap Third Floor Van Vleet, Bldg. Memphis, TN? 38163 (901) 523-9700? ext. 102? (office) (901) 523-9772? (fax) tcostello@gtxinc.com ? ? ____________________________________________________________________________________________________________________ ? This electronic message, including any attachments, is confidential and proprietary and is soley for the intended recipient.? If you are not the intended recipient, this message was sent to you in error and you are hereby advised that any review, disclosure, copying, distribution or use of this message, or any of the information included therein, is unauthorized and strictly prohibited.? If you have received this electronis transmission in error, please immediately notify the sender by reply and permanently delete all copies of this message and its attachments. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dave Johnson Sent: Thursday, August 25, 2005 1:54 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Please help us name our new product Mercedes Medical is coming out with a new teflon coated microtome blade, but we need a name! We have 3 names up for consideration: 1. Advant-Edge 2. Cutting Edge 3. Platinum Blade. Let us know which you like, or come up with a new one and if we use it we'll send you a great gift. Also, come see us at our island booth at NSH in Ft. Lauderdale booth #229. Enter to win a 26" Flat screen TV, and pick up your free Henry the Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be held at the Annual Symposium/Convention from September 10-14 in Ft Lauderdale. You can register on line at www.NSH.org. Visit us on line at www.MercedesMedical.com and get 10% off your first order. Or call at 800.331.2716. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Thu Aug 25 15:36:45 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017175C0@lsexch.lsmaster.lifespan.org> To my way of thinking, "levels" is actually a subset of "recuts", or a specific kind of recuts. "Recuts" means any additional sections taken from a block subsequent to the original sectioning of that block. "Levels" (also sometimes referred to as "step sections") refers to a particular kind of recuts, just as you defined the term. Recuts might include just a given number of sections off the top, or step sections, or every second section, or deeper sections, or serial sections, or mirror image sections, or whatever. But in the everyday jargon of the histology lab, "recuts" and "levels" are usually used just as you described. If you never heard of mirror image sections, don't fret, I didn't either until three weeks ago. One doctor requested that two successive sections be taken, but the first one be placed on the water bath upside down, so the two sections looked like mirror images of each other. What he wanted was sections of the exact same cells at the staining surface of both sections. What will they think of next? > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Patricia Karlisch > Sent: Thursday, August 25, 2005 1:07 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Levels and Recuts > > Hello all, > Can anyone reference a description for 'levels' vs 'recuts'. We use > the terms to mean specific procedures in cutting. Thus, RECUTS are > sections directly off the top of a previously cut block. Levels are > 'recuts' but at different distances between sections. Generally, when > we cut levels we toss the unwanted sections out. In some cases the > unstained are kept. Does anyone in histology consider these to be > 'routine' terms and is this defined in any textbook? > I appreciate your input on this rather basic question. > > > Pat Karlisch > Supervisor, Histology, Pathology and Laboratory Medicine > Penn State Milton S. Hershey Medical Center > Mail Code H179 > Hershey, PA 17033 > Phone (717) 531-6072 > Fax: (717) 531- 7741 > email: pkarlisch@psu.edu > > *****E-Mail Confidentiality Notice***** > This message (including any attachments) contains information intended > for a specific individual(s) and purpose that may be privileged, > confidential or otherwise protected from disclosure pursuant to > applicable law. Any inappropriate use, distribution or copying of the > message is strictly prohibited and may subject you to criminal or civil > penalty. If you have received this transmission in error, please reply > to the sender indicating this error and delete the transmission from > your system immediately. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From froyer <@t> bitstream.net Thu Aug 25 15:37:28 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Please help us name our new product In-Reply-To: <9E8CB17DF8638A46832960D5E3DCAACDC83330@gtxmail1.gtx.gtxinc.com> Message-ID: ..."AmpuQuick", "DigitDemise", "Chopitoff", "Lacero de Pollex".... (Sorry Dave...) Ford ;-) Ford M. Royer, MT(ASCP) Sales Manager - Histology Div. Minnesota Medical Specialists, Inc. 7177 Madison Ave. W. Golden Valley, MN 55427-3601 (888) 790-9686 Fax: 763-546-4830 email: -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Terry Costello Sent: Thursday, August 25, 2005 3:17 PM To: Dave Johnson; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Please help us name our new product "MERCEDES BLADZ" Terrence Costello GTx, Inc. Three North Dunlap Third Floor Van Vleet, Bldg. Memphis, TN? 38163 (901) 523-9700? ext. 102? (office) (901) 523-9772? (fax) tcostello@gtxinc.com ? ? ____________________________________________________________________________ ________________________________________ ? This electronic message, including any attachments, is confidential and proprietary and is soley for the intended recipient.? If you are not the intended recipient, this message was sent to you in error and you are hereby advised that any review, disclosure, copying, distribution or use of this message, or any of the information included therein, is unauthorized and strictly prohibited.? If you have received this electronis transmission in error, please immediately notify the sender by reply and permanently delete all copies of this message and its attachments. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dave Johnson Sent: Thursday, August 25, 2005 1:54 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Please help us name our new product Mercedes Medical is coming out with a new teflon coated microtome blade, but we need a name! We have 3 names up for consideration: 1. Advant-Edge 2. Cutting Edge 3. Platinum Blade. Let us know which you like, or come up with a new one and if we use it we'll send you a great gift. Also, come see us at our island booth at NSH in Ft. Lauderdale booth #229. Enter to win a 26" Flat screen TV, and pick up your free Henry the Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be held at the Annual Symposium/Convention from September 10-14 in Ft Lauderdale. You can register on line at www.NSH.org. Visit us on line at www.MercedesMedical.com and get 10% off your first order. Or call at 800.331.2716. _______________________________________________ Histonet mailing list Histonet@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 petepath <@t> yahoo.com Thu Aug 25 15:45:17 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts Message-ID: <20050825204518.96116.qmail@web30413.mail.mud.yahoo.com> In my training "levels" were " step sections" going a given number of turns of the wheel between each section. A recut was another slide made trimming as little as possible. I have heard people refer to recuts as I described "levels". I think this leads to confusion. I do not know what the official definitions are but if there is a vote, I like it the way I described. From vbaker60 <@t> yahoo.com Thu Aug 25 16:11:24 2005 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Please help us name our new product In-Reply-To: Message-ID: <20050825211124.46276.qmail@web52507.mail.yahoo.com> "Bobbitblade"? Vikki Baker --- Ford Royer wrote: > > ..."AmpuQuick", "DigitDemise", "Chopitoff", "Lacero > de Pollex".... > > (Sorry Dave...) > Ford > ;-) > > Ford M. Royer, MT(ASCP) > Sales Manager - Histology Div. > Minnesota Medical Specialists, Inc. > 7177 Madison Ave. W. > Golden Valley, MN 55427-3601 > (888) 790-9686 > Fax: 763-546-4830 > email: > > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On > Behalf Of Terry > Costello > Sent: Thursday, August 25, 2005 3:17 PM > To: Dave Johnson; Histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Please help us name our new > product > > > "MERCEDES BLADZ" > > Terrence Costello > GTx, Inc. > Three North Dunlap > Third Floor > Van Vleet, Bldg. > Memphis, TN? 38163 > (901) 523-9700? ext. 102? (office) > (901) 523-9772? (fax) > tcostello@gtxinc.com > ? > ? > ____________________________________________________________________________ > ________________________________________ > ? > This electronic message, including any attachments, > is confidential and > proprietary and is soley for the intended > recipient.? If you are not the > intended recipient, this message was sent to you in > error and you are hereby > advised that any review, disclosure, copying, > distribution or use of this > message, or any of the information included therein, > is unauthorized and > strictly prohibited.? If you have received this > electronis transmission in > error, please immediately notify the sender by reply > and permanently delete > all copies of this message and its attachments. > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Dave Johnson > Sent: Thursday, August 25, 2005 1:54 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Please help us name our new > product > > Mercedes Medical is coming out with a new teflon > coated microtome blade, but > we need a name! We have 3 names up for > consideration: > 1. Advant-Edge > 2. Cutting Edge > 3. Platinum Blade. > > Let us know which you like, or come up with a new > one and if we use it we'll > send you a great gift. > > Also, come see us at our island booth at NSH in Ft. > Lauderdale booth #229. > > > Enter to win a 26" Flat screen TV, and pick up your > free Henry the > Histo-Potamus T-shirt and new Histology Catalog. The > NSH show will be held > at the Annual Symposium/Convention from September > 10-14 in Ft Lauderdale. > You can register on line at www.NSH.org. > > > Visit us on line at www.MercedesMedical.com and get > 10% off your first > order. Or call at 800.331.2716. > > > > _______________________________________________ > Histonet mailing list > Histonet@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 > ____________________________________________________ Start your day with Yahoo! - make it your home page http://www.yahoo.com/r/hs From gcallis <@t> montana.edu Thu Aug 25 16:26:59 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts In-Reply-To: <20050825204518.96116.qmail@web30413.mail.mud.yahoo.com> References: <20050825204518.96116.qmail@web30413.mail.mud.yahoo.com> Message-ID: <6.0.0.22.1.20050825152345.01b19008@gemini.msu.montana.edu> Simple concise explanation and I vote yes!! t 02:45 PM 8/25/2005, you wrote: >In my training "levels" were " step sections" going a given number of >turns of the wheel between each section. A recut was another slide made >trimming as little as possible. >I have heard people refer to recuts as I described "levels". I think this >leads to confusion. I >do not know what the official definitions are but if there is a vote, I >like it the way I described. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From sjchtascp <@t> yahoo.com Thu Aug 25 16:35:09 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] rodent muscle processing Message-ID: <20050825213509.15232.qmail@web90210.mail.scd.yahoo.com> I processed several pieces of rodent muscle on a fairly light processing schedual 30-45 minutes per station with 35C. The muscle cuts like it has calcium deposits in it. Is 30-45 minutes in the 1st alcohol (50%) long enough to remove the formalin buffers? I'm thinking this is the problem. Steve --------------------------------- Start your day with Yahoo! - make it your home page From Linke_Noelle <@t> Allergan.com Thu Aug 25 16:39:12 2005 From: Linke_Noelle <@t> Allergan.com (Linke_Noelle) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Please help us name our new product Message-ID: "We Cut You Deep" "Blade 911" Buy 10, get a free prosthetic finger..... -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristen Broomall Sent: Thursday, August 25, 2005 12:16 PM To: 'Dave Johnson'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Please help us name our new product Advant Edge is the name of nutritional products. http://www.easadvantedge.com/adv_products.asp Kristen -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Dave Johnson Sent: Thursday, August 25, 2005 2:54 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Please help us name our new product Mercedes Medical is coming out with a new teflon coated microtome blade, but we need a name! We have 3 names up for consideration: 1. Advant-Edge 2. Cutting Edge 3. Platinum Blade. Let us know which you like, or come up with a new one and if we use it we'll send you a great gift. Also, come see us at our island booth at NSH in Ft. Lauderdale booth #229. Enter to win a 26" Flat screen TV, and pick up your free Henry the Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be held at the Annual Symposium/Convention from September 10-14 in Ft Lauderdale. You can register on line at www.NSH.org. Visit us on line at www.MercedesMedical.com and get 10% off your first order. Or call at 800.331.2716. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> cdc.gov Thu Aug 25 17:25:53 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Microwave Tissue Processors Message-ID: Patsy: I went to Miami to see the Sakura processor in action and processed blocks to bring back with me. The IHC performed on these were fine. I also personally have a Milestone TT/Mega and did a project for a college internship where I evaluated various tissue types processed on this machine and tested them with various antibodies using our established IHC protocols. No only did they stain fine, one of them (our anthrax antibody) actually stained better than when processed by our traditional methods. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Patsy Ruegg Sent: Thu 8/25/2005 3:45 PM To: 'Lori Richey'; 'Joyce Friedland' Cc: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Microwave Tissue Processors I am interested in feed back on IHC results when using MW processors for tissue. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lori Richey Sent: Thursday, August 25, 2005 10:15 AM To: Joyce Friedland Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Microwave Tissue Processors We are unsing the Sakura express microwave processor. We process and cut sm biopsies throughout the day. It works especially well for rush specimens. It can be loaded every 15 minutes. We are still processing derms and breast bxs on our VIPs. Like all new stains and procedures, it takes some time for the pathologists to get use to the different process. Joyce Friedland wrote: > Hi All, > My lab director is interested in purchasing a processor which will > allow us to process batches of cases throughout the day, and serve as > a backup to our two VIPs. > I would greatly appreciate knowing what experiences you all have had > with the various microwave processors available. I have particular > interest in the Pathos from Milestone since it is automated, but I am > open to all possibilities. > > Thanks, > Joyce Friedland > > > _______________________________________________ > Histonet mailing list > Histonet@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 gangli.phd <@t> gmail.com Thu Aug 25 17:17:47 2005 From: gangli.phd <@t> gmail.com (Gang Li) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Please help us name our new product In-Reply-To: <120b7bf050825151433538349@mail.gmail.com> References: <120b7bf050825151433538349@mail.gmail.com> Message-ID: <120b7bf050825151766cc49ff@mail.gmail.com> TCP-Edge TCP-Blade TCP for "Teflon coated platinum" On 8/25/05, Gang Li wrote: > > TCP-Edge or TCP-Blade > > TCP for "Teflon coated platinum" > > > On 8/25/05, Dave Johnson wrote: > > > > Mercedes Medical is coming out with a new teflon coated microtome blade, > > but > > we need a name! We have 3 names up for consideration: > > 1. Advant-Edge > > 2. Cutting Edge > > 3. Platinum Blade. > > > > Let us know which you like, or come up with a new one and if we use it > > we'll > > send you a great gift. > > > > Also, come see us at our island booth at NSH in Ft. Lauderdale booth > > #229. > > > > > > Enter to win a 26" Flat screen TV, and pick up your free Henry the > > Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be > > held > > at the Annual Symposium/Convention from September 10-14 in Ft > > Lauderdale. > > You can register on line at www.NSH.org . > > > > > > Visit us on line at www.MercedesMedical.comand get 10% off your first > > order. Or call at 800.331.2716. > > > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From Eric <@t> ategra.com Thu Aug 25 10:39:17 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Histo Tech Supervisors, and Bench techs Needed for Immediate openings Message-ID: Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. (I have a few travel/temp HistoTech positions as well - see below). Most of my Histo Techs positions are clinical, although I do have a few Research Histo Tech positions as well. HISTOTECH SUPERVISORS: I have positions in Colorado, Oregon & California and who are seeking Histotech Supervisors. BENCH HISTO TECHS: I also have Histo bench positions in Colorado,California,Pennsylvania,Illinois,New Hampshire,West Virginia, Michigan,Oregon,Georgia,Washington,Florida,Minnesota,Ohio and Massachusetts. TRAVEL/TEMP HISTOTECHS: I have Temp assignment openings in South Florida, and California. RESEARCH HISTO TECHS: There are also Research Histo Tech openings in Massachusetts and Ohio as well The clients are currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recruiter 1-800-466-9919 ext 223 From Stephen.Eyres <@t> sanofi-aventis.com Fri Aug 26 02:41:52 2005 From: Stephen.Eyres <@t> sanofi-aventis.com (Stephen.Eyres@sanofi-aventis.com) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Microwave Tissue Processors Message-ID: Hi Donna, Do did your pathologists not notice any significant differences between traditional and microwave processing? Cheers Steve "Willis, Donna" To: "Joyce Friedland" Sent by: histonet-bounces@lists.utsouth cc: western.edu Subject: RE: [Histonet] Microwave Tissue Processors 25/08/2005 18:05 Joyce, The Pathos will allow you to do all that your are interested in doing on biopsy samples and on tissue cut up to 5mm thick. I have one in my lab. You can batch up to 210 cassettes. I have not worked with the Sakura unit but I know that the thickness of the samples on the unit are limited and batch volumes are smaller. Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Friedland Sent: Thursday, August 25, 2005 11:50 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave Tissue Processors Hi All, My lab director is interested in purchasing a processor which will allow us to process batches of cases throughout the day, and serve as a backup to our two VIPs. I would greatly appreciate knowing what experiences you all have had with the various microwave processors available. I have particular interest in the Pathos from Milestone since it is automated, but I am open to all possibilities. Thanks, Joyce Friedland _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- From Krat18 <@t> aol.com Fri Aug 26 06:04:53 2005 From: Krat18 <@t> aol.com (Krat18@aol.com) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts Message-ID: <13f.1a3c3da1.30405155@aol.com> Just to add a little, at our hospital we refer to "serial sections" as sections which immediately follow each other, as we would use for the tiniest specimens or recuts in which the pathologist wants to see the very next cells. Then the "step sections" would have a prescribed space between them. Karen Raterman From jeb1 <@t> stir.ac.uk Fri Aug 26 06:13:16 2005 From: jeb1 <@t> stir.ac.uk (James Bron) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Please help us name our new product In-Reply-To: References: Message-ID: Hiya Dave, How about "Vorpal Blades" from the ultra-sharp sword used to slay the Jabberwok in Lewis Carroll's poem of the same name? Dr James Bron Research Lecturer Parasitology Laboratory Institute of Aquaculture University of Stirling Scotland FK9 4LA email: jeb1@stir.ac.uk fax : 01786 472133 tel. : 01786 467925 ................... ...=-/~~.v.~~\-=... ....(...o.o...).... ...(.\......./.)... ...(./.......\.)... ....(.........).... .....\\\...///..... ......./...\....... ......(.....)...... .......U...U....... .......*I.I*....... .......*I.I*....... .......*.V.*....... .......*...*....... ................... -- The University of Stirling is a university established in Scotland by charter at Stirling, FK9 4LA. Privileged/Confidential Information may be contained in this message. If you are not the addressee indicated in this message (or responsible for delivery of the message to such person), you may not disclose, copy or deliver this message to anyone and any action taken or omitted to be taken in reliance on it, is prohibited and may be unlawful. In such case, you should destroy this message and kindly notify the sender by reply email. Please advise immediately if you or your employer do not consent to Internet email for messages of this kind. From Terry.Marshall <@t> rothgen.nhs.uk Fri Aug 26 06:29:41 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts Message-ID: Another subject talked about several times before. I agree it is sensible to call another section (usually 'cos there is something technically wrong with the first), a recut. I usually scribble on the form, something like, "bloody rubbish" or "?Q.C.", or something similarly inflammatory:-) Levels are just a few extra sections at an arbitrary interval. Serial sections are self explanatory, and semi-serial is a combination of serial and levels - cut in a bit then do a burst of serials, then repeat. Sometimes I ask for levels through the block. What is asked for depends and what you are looking for and how you expect to find it. Invariably, for anything other than levels, I talk about the problem to the tech., so that they can cut with an eye to the problem. 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: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 25 August 2005 21:45 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Levels and Recuts In my training "levels" were " step sections" going a given number of turns of the wheel between each section. A recut was another slide made trimming as little as possible. I have heard people refer to recuts as I described "levels". I think this leads to confusion. I do not know what the official definitions are but if there is a vote, I like it the way I described. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> cdc.gov Fri Aug 26 06:32:21 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts Message-ID: "Invariably, for anything other than levels, I talk about the problem to the tech., so that they can cut with an eye to the problem." Discuss it with the tech......Gee, what a concept! :) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Friday, August 26, 2005 7:30 AM To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Another subject talked about several times before. I agree it is sensible to call another section (usually 'cos there is something technically wrong with the first), a recut. I usually scribble on the form, something like, "bloody rubbish" or "?Q.C.", or something similarly inflammatory:-) Levels are just a few extra sections at an arbitrary interval. Serial sections are self explanatory, and semi-serial is a combination of serial and levels - cut in a bit then do a burst of serials, then repeat. Sometimes I ask for levels through the block. What is asked for depends and what you are looking for and how you expect to find it. Invariably, for anything other than levels, I talk about the problem to the tech., so that they can cut with an eye to the problem. 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: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 25 August 2005 21:45 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Levels and Recuts In my training "levels" were " step sections" going a given number of turns of the wheel between each section. A recut was another slide made trimming as little as possible. I have heard people refer to recuts as I described "levels". I think this leads to confusion. I do not know what the official definitions are but if there is a vote, I like it the way I described. _______________________________________________ Histonet mailing list Histonet@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 nancy.troiano <@t> yale.edu Fri Aug 26 06:38:58 2005 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] EDTA recipe with PVP Message-ID: <5.2.1.1.2.20050826073656.00c11d08@email.med.yale.edu> Here is our recipe for EDTA containing PVP: Dissolve 41.3 grams EDTA (disodium salt), 4.4 grams Sodium Hydroxide and 50 g PVP-40 (Sigma) in 1000 ml distilled water over very low heat. Cool to room temperature, adjust pH to 7.4 once cooled. We use this to gently decalcify our mouse long bones, it will take about 3 weeks to decalcify an adult mouse femur. From Kemlo.Rogerson <@t> elht.nhs.uk Fri Aug 26 06:40:07 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD50D@elht-exch1.xelht.nhs.uk> "bloody rubbish"? Is that accepted terminology for one Professional to talk to another? Only teasing but it does highlight one worrying aspect in some Histology Labs. If Biomedical Scientists are really going to be respected by their Medical colleagues then they have to accept responsibility for the work they produce. If a Transfusion BMS gets it wrong the Patient gets maybe the wrong blood type; if a Histology BMS gets it wrong, he/she gets told it's 'bloody rubbish'. If the Medic gets it wrong then it has an effect on Patient management and/ or their can be litigation. I'm all for extending the role of the BMS in Histology, but from my experience, and Terry's response we must do what we do now, better, it seems. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 26 August 2005 12:30 To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Another subject talked about several times before. I agree it is sensible to call another section (usually 'cos there is something technically wrong with the first), a recut. I usually scribble on the form, something like, "bloody rubbish" or "?Q.C.", or something similarly inflammatory:-) Levels are just a few extra sections at an arbitrary interval. Serial sections are self explanatory, and semi-serial is a combination of serial and levels - cut in a bit then do a burst of serials, then repeat. Sometimes I ask for levels through the block. What is asked for depends and what you are looking for and how you expect to find it. Invariably, for anything other than levels, I talk about the problem to the tech., so that they can cut with an eye to the problem. 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: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 25 August 2005 21:45 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Levels and Recuts In my training "levels" were " step sections" going a given number of turns of the wheel between each section. A recut was another slide made trimming as little as possible. I have heard people refer to recuts as I described "levels". I think this leads to confusion. I do not know what the official definitions are but if there is a vote, I like it the way I described. _______________________________________________ Histonet mailing list Histonet@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 Fri Aug 26 07:13:56 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts Message-ID: Well, I do often write that stuff, but it's good natured teasing too. Seriously, one intractable problem in histology is that the tech. produces and the pathologist interprets. Ideally it should be done by the same people (or even) person. It just isn't possible. 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: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 26 August 2005 12:40 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts "bloody rubbish"? Is that accepted terminology for one Professional to talk to another? Only teasing but it does highlight one worrying aspect in some Histology Labs. If Biomedical Scientists are really going to be respected by their Medical colleagues then they have to accept responsibility for the work they produce. If a Transfusion BMS gets it wrong the Patient gets maybe the wrong blood type; if a Histology BMS gets it wrong, he/she gets told it's 'bloody rubbish'. If the Medic gets it wrong then it has an effect on Patient management and/ or their can be litigation. I'm all for extending the role of the BMS in Histology, but from my experience, and Terry's response we must do what we do now, better, it seems. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 26 August 2005 12:30 To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Another subject talked about several times before. I agree it is sensible to call another section (usually 'cos there is something technically wrong with the first), a recut. I usually scribble on the form, something like, "bloody rubbish" or "?Q.C.", or something similarly inflammatory:-) Levels are just a few extra sections at an arbitrary interval. Serial sections are self explanatory, and semi-serial is a combination of serial and levels - cut in a bit then do a burst of serials, then repeat. Sometimes I ask for levels through the block. What is asked for depends and what you are looking for and how you expect to find it. Invariably, for anything other than levels, I talk about the problem to the tech., so that they can cut with an eye to the problem. 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: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 25 August 2005 21:45 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Levels and Recuts In my training "levels" were " step sections" going a given number of turns of the wheel between each section. A recut was another slide made trimming as little as possible. I have heard people refer to recuts as I described "levels". I think this leads to confusion. I do not know what the official definitions are but if there is a vote, I like it the way I described. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Fri Aug 26 07:28:23 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD50E@elht-exch1.xelht.nhs.uk> Can't it? Really why not? Are you seriously saying that a properly qualified and trained BMS is unable to report on such things as appendix, gall bladder, vas deferens, etc. Surely not as that defies the very 80:20 rule you so generously taught me. The most important aspect of reporting that a BMS has to learn is the boundary of his/ her training. If something is without that boundary then referral to a Medic is needed; the old chestnut of 'most of these are easy to report, but you need medical training to identify a subtle change' does not hold water; surely experience and training is the real secret. If something is present that ought not be, then Cytology has shown us that the BMS is just a likely to see it, but I concede may not have the skill to report it. Are you really going to wade through shed loads of gut biopsies to weed out the negatives from the abnormals in colorectal cancer screening? Is that a good use of your valuable and expensive time? The other benefit will be, as in cytology, if the slide is 'bloody rubbish' then that will motivate the BMS to do the job better as he or she will have to report some of them; nothing like having your name on a report to crystallise the mind. You were always very good at 'thinking out of the box', but I expect you were counting on a reply like this. -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 26 August 2005 13:14 To: Rogerson Kemlo (ELHT) Pathology; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Well, I do often write that stuff, but it's good natured teasing too. Seriously, one intractable problem in histology is that the tech. produces and the pathologist interprets. Ideally it should be done by the same people (or even) person. It just isn't possible. 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: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 26 August 2005 12:40 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts "bloody rubbish"? Is that accepted terminology for one Professional to talk to another? Only teasing but it does highlight one worrying aspect in some Histology Labs. If Biomedical Scientists are really going to be respected by their Medical colleagues then they have to accept responsibility for the work they produce. If a Transfusion BMS gets it wrong the Patient gets maybe the wrong blood type; if a Histology BMS gets it wrong, he/she gets told it's 'bloody rubbish'. If the Medic gets it wrong then it has an effect on Patient management and/ or their can be litigation. I'm all for extending the role of the BMS in Histology, but from my experience, and Terry's response we must do what we do now, better, it seems. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 26 August 2005 12:30 To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Another subject talked about several times before. I agree it is sensible to call another section (usually 'cos there is something technically wrong with the first), a recut. I usually scribble on the form, something like, "bloody rubbish" or "?Q.C.", or something similarly inflammatory:-) Levels are just a few extra sections at an arbitrary interval. Serial sections are self explanatory, and semi-serial is a combination of serial and levels - cut in a bit then do a burst of serials, then repeat. Sometimes I ask for levels through the block. What is asked for depends and what you are looking for and how you expect to find it. Invariably, for anything other than levels, I talk about the problem to the tech., so that they can cut with an eye to the problem. 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: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 25 August 2005 21:45 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Levels and Recuts In my training "levels" were " step sections" going a given number of turns of the wheel between each section. A recut was another slide made trimming as little as possible. I have heard people refer to recuts as I described "levels". I think this leads to confusion. I do not know what the official definitions are but if there is a vote, I like it the way I described. _______________________________________________ Histonet mailing list Histonet@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 MSafron <@t> wilresearch.com Fri Aug 26 07:32:03 2005 From: MSafron <@t> wilresearch.com (Mike Safron) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] rodent muscle processing Message-ID: Steve, After processing and embedding, how are you preparing the tissue for sectioning (soaking, icing, etc..) I don't think the formalin buffers are your problem. Our rat processing schedule starts with 30 minutes in 70% Alcohol and we have no problem. Michael Safron A.S.,HT(ASCP) Manager, Histology WIL Research Laboratories LLC Ashland, Ohio 44805 Msafron@wilresearch.com From Terry.Marshall <@t> rothgen.nhs.uk Fri Aug 26 07:37:08 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts Message-ID: Kemlo, I was not saying, seriously or otherwise. I did not use the word "unable". However, I will now say it. Not unable due to deficiency of ability of course, but to practicalities of licensing etc. What I was thinking of when I said "It just isn't possible.", is the "most efficient use of time" element. For a start, to be in to block out and cut, I would have to get up early in the morning, and you may remember how difficult I found that:-) 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: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 26 August 2005 13:28 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Can't it? Really why not? Are you seriously saying that a properly qualified and trained BMS is unable to report on such things as appendix, gall bladder, vas deferens, etc. Surely not as that defies the very 80:20 rule you so generously taught me. The most important aspect of reporting that a BMS has to learn is the boundary of his/ her training. If something is without that boundary then referral to a Medic is needed; the old chestnut of 'most of these are easy to report, but you need medical training to identify a subtle change' does not hold water; surely experience and training is the real secret. If something is present that ought not be, then Cytology has shown us that the BMS is just a likely to see it, but I concede may not have the skill to report it. Are you really going to wade through shed loads of gut biopsies to weed out the negatives from the abnormals in colorectal cancer screening? Is that a good use of your valuable and expensive time? The other benefit will be, as in cytology, if the slide is 'bloody rubbish' then that will motivate the BMS to do the job better as he or she will have to report some of them; nothing like having your name on a report to crystallise the mind. You were always very good at 'thinking out of the box', but I expect you were counting on a reply like this. -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 26 August 2005 13:14 To: Rogerson Kemlo (ELHT) Pathology; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Well, I do often write that stuff, but it's good natured teasing too. Seriously, one intractable problem in histology is that the tech. produces and the pathologist interprets. Ideally it should be done by the same people (or even) person. It just isn't possible. 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: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 26 August 2005 12:40 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts "bloody rubbish"? Is that accepted terminology for one Professional to talk to another? Only teasing but it does highlight one worrying aspect in some Histology Labs. If Biomedical Scientists are really going to be respected by their Medical colleagues then they have to accept responsibility for the work they produce. If a Transfusion BMS gets it wrong the Patient gets maybe the wrong blood type; if a Histology BMS gets it wrong, he/she gets told it's 'bloody rubbish'. If the Medic gets it wrong then it has an effect on Patient management and/ or their can be litigation. I'm all for extending the role of the BMS in Histology, but from my experience, and Terry's response we must do what we do now, better, it seems. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 26 August 2005 12:30 To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Another subject talked about several times before. I agree it is sensible to call another section (usually 'cos there is something technically wrong with the first), a recut. I usually scribble on the form, something like, "bloody rubbish" or "?Q.C.", or something similarly inflammatory:-) Levels are just a few extra sections at an arbitrary interval. Serial sections are self explanatory, and semi-serial is a combination of serial and levels - cut in a bit then do a burst of serials, then repeat. Sometimes I ask for levels through the block. What is asked for depends and what you are looking for and how you expect to find it. Invariably, for anything other than levels, I talk about the problem to the tech., so that they can cut with an eye to the problem. 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: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 25 August 2005 21:45 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Levels and Recuts In my training "levels" were " step sections" going a given number of turns of the wheel between each section. A recut was another slide made trimming as little as possible. I have heard people refer to recuts as I described "levels". I think this leads to confusion. I do not know what the official definitions are but if there is a vote, I like it the way I described. _______________________________________________ Histonet mailing list Histonet@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 Delventh3 <@t> aol.com Fri Aug 26 07:51:01 2005 From: Delventh3 <@t> aol.com (Delventh3@aol.com) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] The Mercedes Knife Message-ID: <1e9.42f565e7.30406a35@aol.com> How about calling it the "Mercedes Edge" Hello everyone - Yes I'm still out here in Histo Land. I work whenever a short assignment comes up. So far have been to Dallas, Tx and Coral Gables, Fl. this year. We've moved to Texas and are enjoying the sun shine and heat. Take care--Always informative to read the HistoNet. Priscilla D. From Kemlo.Rogerson <@t> elht.nhs.uk Fri Aug 26 07:52:42 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD50F@elht-exch1.xelht.nhs.uk> I accept what you say, but it wouldn't work like that would it? Remember the very small team of 'skin' Techs (1) you once had? I remember that team had enough time to cut up (gross), embed, stain, Q/C and then go through the slides before you had them and take a stab of interpreting them. You were generous enough to teach the team (of 1) the bsic rudiments of Dermatopathology; that art could well be deemed to be outside the realms of a BMS, but I certainly had enough time to report them if I had the skill. The practicalities of licensing, I would respectfully suggest, is not an issue; as long as training is adequate and revisited and the proper piece of paper gained, then it's not an issue. We could start with BMS reporting the ICC couldn't we? Granted it needs good communication our Medical colleagues but us Techs are getting the hang being loquacious. -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 26 August 2005 13:37 To: Rogerson Kemlo (ELHT) Pathology; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Kemlo, I was not saying, seriously or otherwise. I did not use the word "unable". However, I will now say it. Not unable due to deficiency of ability of course, but to practicalities of licensing etc. What I was thinking of when I said "It just isn't possible.", is the "most efficient use of time" element. For a start, to be in to block out and cut, I would have to get up early in the morning, and you may remember how difficult I found that:-) 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: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 26 August 2005 13:28 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Can't it? Really why not? Are you seriously saying that a properly qualified and trained BMS is unable to report on such things as appendix, gall bladder, vas deferens, etc. Surely not as that defies the very 80:20 rule you so generously taught me. The most important aspect of reporting that a BMS has to learn is the boundary of his/ her training. If something is without that boundary then referral to a Medic is needed; the old chestnut of 'most of these are easy to report, but you need medical training to identify a subtle change' does not hold water; surely experience and training is the real secret. If something is present that ought not be, then Cytology has shown us that the BMS is just a likely to see it, but I concede may not have the skill to report it. Are you really going to wade through shed loads of gut biopsies to weed out the negatives from the abnormals in colorectal cancer screening? Is that a good use of your valuable and expensive time? The other benefit will be, as in cytology, if the slide is 'bloody rubbish' then that will motivate the BMS to do the job better as he or she will have to report some of them; nothing like having your name on a report to crystallise the mind. You were always very good at 'thinking out of the box', but I expect you were counting on a reply like this. -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 26 August 2005 13:14 To: Rogerson Kemlo (ELHT) Pathology; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Well, I do often write that stuff, but it's good natured teasing too. Seriously, one intractable problem in histology is that the tech. produces and the pathologist interprets. Ideally it should be done by the same people (or even) person. It just isn't possible. 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: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 26 August 2005 12:40 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts "bloody rubbish"? Is that accepted terminology for one Professional to talk to another? Only teasing but it does highlight one worrying aspect in some Histology Labs. If Biomedical Scientists are really going to be respected by their Medical colleagues then they have to accept responsibility for the work they produce. If a Transfusion BMS gets it wrong the Patient gets maybe the wrong blood type; if a Histology BMS gets it wrong, he/she gets told it's 'bloody rubbish'. If the Medic gets it wrong then it has an effect on Patient management and/ or their can be litigation. I'm all for extending the role of the BMS in Histology, but from my experience, and Terry's response we must do what we do now, better, it seems. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 26 August 2005 12:30 To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Another subject talked about several times before. I agree it is sensible to call another section (usually 'cos there is something technically wrong with the first), a recut. I usually scribble on the form, something like, "bloody rubbish" or "?Q.C.", or something similarly inflammatory:-) Levels are just a few extra sections at an arbitrary interval. Serial sections are self explanatory, and semi-serial is a combination of serial and levels - cut in a bit then do a burst of serials, then repeat. Sometimes I ask for levels through the block. What is asked for depends and what you are looking for and how you expect to find it. Invariably, for anything other than levels, I talk about the problem to the tech., so that they can cut with an eye to the problem. 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: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 25 August 2005 21:45 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Levels and Recuts In my training "levels" were " step sections" going a given number of turns of the wheel between each section. A recut was another slide made trimming as little as possible. I have heard people refer to recuts as I described "levels". I think this leads to confusion. I do not know what the official definitions are but if there is a vote, I like it the way I described. _______________________________________________ Histonet mailing list Histonet@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 DonnaWillis <@t> texashealth.org Fri Aug 26 07:53:17 2005 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Microwave Tissue Processors Message-ID: <4483B6EE0FA82A4EAED2B1E161E5E41147C9BE@ftwex02.txhealth.org> No difference with the chemicals that we use. Donna -----Original Message----- From: Stephen.Eyres@sanofi-aventis.com [mailto:Stephen.Eyres@sanofi-aventis.com] Sent: Friday, August 26, 2005 2:42 AM To: Willis, Donna Cc: Histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu; Joyce Friedland Subject: RE: [Histonet] Microwave Tissue Processors Hi Donna, Do did your pathologists not notice any significant differences between traditional and microwave processing? Cheers Steve "Willis, Donna" To: "Joyce Friedland" Sent by: histonet-bounces@lists.utsouth cc: western.edu Subject: RE: [Histonet] Microwave Tissue Processors 25/08/2005 18:05 Joyce, The Pathos will allow you to do all that your are interested in doing on biopsy samples and on tissue cut up to 5mm thick. I have one in my lab. You can batch up to 210 cassettes. I have not worked with the Sakura unit but I know that the thickness of the samples on the unit are limited and batch volumes are smaller. Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joyce Friedland Sent: Thursday, August 25, 2005 11:50 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Microwave Tissue Processors Hi All, My lab director is interested in purchasing a processor which will allow us to process batches of cases throughout the day, and serve as a backup to our two VIPs. I would greatly appreciate knowing what experiences you all have had with the various microwave processors available. I have particular interest in the Pathos from Milestone since it is automated, but I am open to all possibilities. Thanks, Joyce Friedland _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- 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 Kemlo.Rogerson <@t> elht.nhs.uk Fri Aug 26 08:06:46 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Microwave Tissue Processors Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E698F21@elht-exch1.xelht.nhs.uk> How do you brown sausages? Kemlo If you go flying back through time and you see somebody else flying forward into the future, it's probably best to avoid eye contact. Hi Donna, Do did your pathologists not notice any significant differences between traditional and microwave processing? Cheers Steve Joyce, From Terry.Marshall <@t> rothgen.nhs.uk Fri Aug 26 08:07:18 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Levels and Recuts Message-ID: I've no idea what ICC is, but yes:-) All this is possible in time, and given a receptive attitude of those in control. Ah. There's the stumbling block. We've strayed a bit from the subject of levels and recuts:-) 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: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 26 August 2005 13:53 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts I accept what you say, but it wouldn't work like that would it? Remember the very small team of 'skin' Techs (1) you once had? I remember that team had enough time to cut up (gross), embed, stain, Q/C and then go through the slides before you had them and take a stab of interpreting them. You were generous enough to teach the team (of 1) the bsic rudiments of Dermatopathology; that art could well be deemed to be outside the realms of a BMS, but I certainly had enough time to report them if I had the skill. The practicalities of licensing, I would respectfully suggest, is not an issue; as long as training is adequate and revisited and the proper piece of paper gained, then it's not an issue. We could start with BMS reporting the ICC couldn't we? Granted it needs good communication our Medical colleagues but us Techs are getting the hang being loquacious. -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 26 August 2005 13:37 To: Rogerson Kemlo (ELHT) Pathology; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Kemlo, I was not saying, seriously or otherwise. I did not use the word "unable". However, I will now say it. Not unable due to deficiency of ability of course, but to practicalities of licensing etc. What I was thinking of when I said "It just isn't possible.", is the "most efficient use of time" element. For a start, to be in to block out and cut, I would have to get up early in the morning, and you may remember how difficult I found that:-) 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: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 26 August 2005 13:28 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Can't it? Really why not? Are you seriously saying that a properly qualified and trained BMS is unable to report on such things as appendix, gall bladder, vas deferens, etc. Surely not as that defies the very 80:20 rule you so generously taught me. The most important aspect of reporting that a BMS has to learn is the boundary of his/ her training. If something is without that boundary then referral to a Medic is needed; the old chestnut of 'most of these are easy to report, but you need medical training to identify a subtle change' does not hold water; surely experience and training is the real secret. If something is present that ought not be, then Cytology has shown us that the BMS is just a likely to see it, but I concede may not have the skill to report it. Are you really going to wade through shed loads of gut biopsies to weed out the negatives from the abnormals in colorectal cancer screening? Is that a good use of your valuable and expensive time? The other benefit will be, as in cytology, if the slide is 'bloody rubbish' then that will motivate the BMS to do the job better as he or she will have to report some of them; nothing like having your name on a report to crystallise the mind. You were always very good at 'thinking out of the box', but I expect you were counting on a reply like this. -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: 26 August 2005 13:14 To: Rogerson Kemlo (ELHT) Pathology; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Well, I do often write that stuff, but it's good natured teasing too. Seriously, one intractable problem in histology is that the tech. produces and the pathologist interprets. Ideally it should be done by the same people (or even) person. It just isn't possible. 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: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: 26 August 2005 12:40 To: Marshall Terry Dr, Consultant Histopathologist; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts "bloody rubbish"? Is that accepted terminology for one Professional to talk to another? Only teasing but it does highlight one worrying aspect in some Histology Labs. If Biomedical Scientists are really going to be respected by their Medical colleagues then they have to accept responsibility for the work they produce. If a Transfusion BMS gets it wrong the Patient gets maybe the wrong blood type; if a Histology BMS gets it wrong, he/she gets told it's 'bloody rubbish'. If the Medic gets it wrong then it has an effect on Patient management and/ or their can be litigation. I'm all for extending the role of the BMS in Histology, but from my experience, and Terry's response we must do what we do now, better, it seems. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: 26 August 2005 12:30 To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Another subject talked about several times before. I agree it is sensible to call another section (usually 'cos there is something technically wrong with the first), a recut. I usually scribble on the form, something like, "bloody rubbish" or "?Q.C.", or something similarly inflammatory:-) Levels are just a few extra sections at an arbitrary interval. Serial sections are self explanatory, and semi-serial is a combination of serial and levels - cut in a bit then do a burst of serials, then repeat. Sometimes I ask for levels through the block. What is asked for depends and what you are looking for and how you expect to find it. Invariably, for anything other than levels, I talk about the problem to the tech., so that they can cut with an eye to the problem. 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: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 25 August 2005 21:45 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Levels and Recuts In my training "levels" were " step sections" going a given number of turns of the wheel between each section. A recut was another slide made trimming as little as possible. I have heard people refer to recuts as I described "levels". I think this leads to confusion. I do not know what the official definitions are but if there is a vote, I like it the way I described. _______________________________________________ Histonet mailing list Histonet@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 dmarsha3 <@t> utmem.edu Fri Aug 26 08:36:25 2005 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Please help us name our new product References: <9E8CB17DF8638A46832960D5E3DCAACDC83330@gtxmail1.gtx.gtxinc.com> Message-ID: <001501c5aa43$277fb9e0$f623c084@DanaM> i like this one! dm ----- Original Message ----- From: "Terry Costello" To: "Dave Johnson" ; Sent: Thursday, August 25, 2005 3:16 PM Subject: RE: [Histonet] Please help us name our new product "MERCEDES BLADZ" Terrence Costello GTx, Inc. Three North Dunlap Third Floor Van Vleet, Bldg. Memphis, TN 38163 (901) 523-9700 ext. 102 (office) (901) 523-9772 (fax) tcostello@gtxinc.com ____________________________________________________________________________________________________________________ This electronic message, including any attachments, is confidential and proprietary and is soley for the intended recipient. If you are not the intended recipient, this message was sent to you in error and you are hereby advised that any review, disclosure, copying, distribution or use of this message, or any of the information included therein, is unauthorized and strictly prohibited. If you have received this electronis transmission in error, please immediately notify the sender by reply and permanently delete all copies of this message and its attachments. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dave Johnson Sent: Thursday, August 25, 2005 1:54 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Please help us name our new product Mercedes Medical is coming out with a new teflon coated microtome blade, but we need a name! We have 3 names up for consideration: 1. Advant-Edge 2. Cutting Edge 3. Platinum Blade. Let us know which you like, or come up with a new one and if we use it we'll send you a great gift. Also, come see us at our island booth at NSH in Ft. Lauderdale booth #229. Enter to win a 26" Flat screen TV, and pick up your free Henry the Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be held at the Annual Symposium/Convention from September 10-14 in Ft Lauderdale. You can register on line at www.NSH.org. Visit us on line at www.MercedesMedical.com and get 10% off your first order. Or call at 800.331.2716. _______________________________________________ Histonet mailing list Histonet@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 Sarah_Mack <@t> urmc.rochester.edu Fri Aug 26 08:51:44 2005 From: Sarah_Mack <@t> urmc.rochester.edu (Mack, Sarah) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Please help us name our new product Message-ID: I agree! Sarah > ---------- > From: Dana Marshall > Sent: Friday, August 26, 2005 9:36 AM > To: Terry Costello; Dave Johnson; Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Please help us name our new product > > i like this one! > dm > ----- Original Message ----- > From: "Terry Costello" > To: "Dave Johnson" ; > > Sent: Thursday, August 25, 2005 3:16 PM > Subject: RE: [Histonet] Please help us name our new product > > > "MERCEDES BLADZ" > > Terrence Costello > GTx, Inc. > Three North Dunlap > Third Floor > Van Vleet, Bldg. > Memphis, TN 38163 > (901) 523-9700 ext. 102 (office) > (901) 523-9772 (fax) > tcostello@gtxinc.com > > > __________________________________________________________________________ > __________________________________________ > > This electronic message, including any attachments, is confidential and > proprietary and is soley for the intended recipient. If you are not the > intended recipient, this message was sent to you in error and you are > hereby > advised that any review, disclosure, copying, distribution or use of this > message, or any of the information included therein, is unauthorized and > strictly prohibited. If you have received this electronis transmission in > error, please immediately notify the sender by reply and permanently > delete > all copies of this message and its attachments. > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dave > Johnson > Sent: Thursday, August 25, 2005 1:54 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Please help us name our new product > > Mercedes Medical is coming out with a new teflon coated microtome blade, > but > we need a name! We have 3 names up for consideration: > 1. Advant-Edge > 2. Cutting Edge > 3. Platinum Blade. > > Let us know which you like, or come up with a new one and if we use it > we'll > send you a great gift. > > Also, come see us at our island booth at NSH in Ft. Lauderdale booth #229. > > > Enter to win a 26" Flat screen TV, and pick up your free Henry the > Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be held > at the Annual Symposium/Convention from September 10-14 in Ft Lauderdale. > You can register on line at www.NSH.org. > > > Visit us on line at www.MercedesMedical.com and get 10% off your first > order. Or call at 800.331.2716. > > > > _______________________________________________ > Histonet mailing list > Histonet@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 Stephen.Eyres <@t> sanofi-aventis.com Fri Aug 26 09:08:04 2005 From: Stephen.Eyres <@t> sanofi-aventis.com (Stephen.Eyres@sanofi-aventis.com) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Please help us name our new product Message-ID: Now look here chaps and chapesses, this has gone on long enough. I don't want to put a BLOCK on this, nor have I a SCORE to settle, and I don't want to WAX lyrical about blades. However, a good name will give you an EDGE over the competition, who, after all, are always trying to STEEL a march on everyone else. A catchy name will make you a CUT above the rest, and in sales terms BEVALuable. On the other hand, ours is only a small SECTION of the market with customers often FLOATING between brands after SOAKING up all the sales blurb. I think there is a need for TRIMMINGIN the blade market, or so a promotional CASSETTE, wrapped in a nice RIBBON, informed me. In fact there were several cassettes, a SERIAL promotion I suppose you could say. I have lost one of them. Someone must have NICKED it. What LEVELS some folk will stoop to. I think I've RAKEed up enough dust now and if I HEIFFOR see any more messages on this subject I could get into a STROP . I've had enough now. Time to go an HONE some of my other skills. Do you think I've gone off my ROCKER, CAMBRIDGE of course. What if my COVERSLIPS? Do you think someone will LABEL me a nutter? I'm getting tired now. In fact I'm CHATTERED. Wish someone would close the VENETIAN BLIND. Think I need a pint, my throats really DRY. Cheers Steve "Dana Marshall" To: "Terry Costello" Sent by: "Dave Johnson" histonet-bounces@lists.utsouth western.edu cc: Subject: Re: [Histonet] Please help us name our new product 26/08/2005 14:36 i like this one! dm ----- Original Message ----- From: "Terry Costello" To: "Dave Johnson" ; Sent: Thursday, August 25, 2005 3:16 PM Subject: RE: [Histonet] Please help us name our new product "MERCEDES BLADZ" Terrence Costello GTx, Inc. Three North Dunlap Third Floor Van Vleet, Bldg. Memphis, TN 38163 (901) 523-9700 ext. 102 (office) (901) 523-9772 (fax) tcostello@gtxinc.com ____________________________________________________________________________________________________________________ This electronic message, including any attachments, is confidential and proprietary and is soley for the intended recipient. If you are not the intended recipient, this message was sent to you in error and you are hereby advised that any review, disclosure, copying, distribution or use of this message, or any of the information included therein, is unauthorized and strictly prohibited. If you have received this electronis transmission in error, please immediately notify the sender by reply and permanently delete all copies of this message and its attachments. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dave Johnson Sent: Thursday, August 25, 2005 1:54 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Please help us name our new product Mercedes Medical is coming out with a new teflon coated microtome blade, but we need a name! We have 3 names up for consideration: 1. Advant-Edge 2. Cutting Edge 3. Platinum Blade. Let us know which you like, or come up with a new one and if we use it we'll send you a great gift. Also, come see us at our island booth at NSH in Ft. Lauderdale booth #229. Enter to win a 26" Flat screen TV, and pick up your free Henry the Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be held at the Annual Symposium/Convention from September 10-14 in Ft Lauderdale. You can register on line at www.NSH.org. Visit us on line at www.MercedesMedical.com and get 10% off your first order. Or call at 800.331.2716. _______________________________________________ Histonet mailing list Histonet@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 ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- From dsnider <@t> shrinenet.org Fri Aug 26 09:46:04 2005 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:25:30 2005 Subject: [Histonet] Procedure for Mallory's Trichrome Stain Message-ID: <84BE46B37B314D409C5A17B7BAB022D616031A@IDC-EX-VS01.shriners.cc> Hello again netters.... I am trying to find a procedure for Mallory's Trichrome and am not having an easy time of it! Once was emailed to me, but it skips an entire step.(one of the reagents in's used at all!) I have done this stain in the past, but its been a while and I cannot remember the procedure at all. Please help! It can be emailed to me or faxed...513-872-6072. The Masson is not demonstarting what the researcher wants to see. Thanks in advance... Deanna Snider HT ASCP Shriners Hospital for Children Research Dept. Cincinnati, Oh 513-872-6388 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 gcallis <@t> montana.edu Fri Aug 26 10:00:49 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:30 2005 Subject: Question about Re: [Histonet] EDTA recipe with PVP In-Reply-To: <5.2.1.1.2.20050826073656.00c11d08@email.med.yale.edu> References: <5.2.1.1.2.20050826073656.00c11d08@email.med.yale.edu> Message-ID: <6.0.0.22.1.20050826085927.01b7cf48@gemini.msu.montana.edu> Nancy, What is the advantage of adding PVP to the EDTA decalcifying solution? Just curious. At 05:38 AM 8/26/2005, you wrote: >Here is our recipe for EDTA containing PVP: Dissolve 41.3 grams EDTA >(disodium salt), 4.4 grams Sodium Hydroxide and 50 g PVP-40 (Sigma) in >1000 ml distilled water over very low heat. Cool to room temperature, >adjust pH to 7.4 once cooled. We use this to gently decalcify our mouse >long bones, it will take about 3 weeks to decalcify an adult mouse femur. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From Tbarnhart <@t> primecare.org Fri Aug 26 10:01:42 2005 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Fri Sep 16 15:25:31 2005 Subject: [BULK] Re: [Histonet] Please help us name our new product Message-ID: <1779904B5E82D511914C00D0B793339205BFD9EB@exchangent> Bravo....Bravo.....lol -----Original Message----- From: Stephen.Eyres@sanofi-aventis.com [mailto:Stephen.Eyres@sanofi-aventis.com] Sent: Friday, August 26, 2005 9:08 AM To: Dana Marshall Cc: Histonet@lists.utsouthwestern.edu; Terry Costello; histonet-bounces@lists.utsouthwestern.edu Subject: [BULK] Re: [Histonet] Please help us name our new product Importance: Low Now look here chaps and chapesses, this has gone on long enough. I don't want to put a BLOCK on this, nor have I a SCORE to settle, and I don't want to WAX lyrical about blades. However, a good name will give you an EDGE over the competition, who, after all, are always trying to STEEL a march on everyone else. A catchy name will make you a CUT above the rest, and in sales terms BEVALuable. On the other hand, ours is only a small SECTION of the market with customers often FLOATING between brands after SOAKING up all the sales blurb. I think there is a need for TRIMMINGIN the blade market, or so a promotional CASSETTE, wrapped in a nice RIBBON, informed me. In fact there were several cassettes, a SERIAL promotion I suppose you could say. I have lost one of them. Someone must have NICKED it. What LEVELS some folk will stoop to. I think I've RAKEed up enough dust now and if I HEIFFOR see any more messages on this subject I could get into a STROP . I've had enough now. Time to go an HONE some of my other skills. Do you think I've gone off my ROCKER, CAMBRIDGE of course. What if my COVERSLIPS? Do you think someone will LABEL me a nutter? I'm getting tired now. In fact I'm CHATTERED. Wish someone would close the VENETIAN BLIND. Think I need a pint, my throats really DRY. Cheers Steve "Dana Marshall" To: "Terry Costello" Sent by: "Dave Johnson" histonet-bounces@lists.utsouth western.edu cc: Subject: Re: [Histonet] Please help us name our new product 26/08/2005 14:36 i like this one! dm ----- Original Message ----- From: "Terry Costello" To: "Dave Johnson" ; Sent: Thursday, August 25, 2005 3:16 PM Subject: RE: [Histonet] Please help us name our new product "MERCEDES BLADZ" Terrence Costello GTx, Inc. Three North Dunlap Third Floor Van Vleet, Bldg. Memphis, TN 38163 (901) 523-9700 ext. 102 (office) (901) 523-9772 (fax) tcostello@gtxinc.com ____________________________________________________________________________ ________________________________________ This electronic message, including any attachments, is confidential and proprietary and is soley for the intended recipient. If you are not the intended recipient, this message was sent to you in error and you are hereby advised that any review, disclosure, copying, distribution or use of this message, or any of the information included therein, is unauthorized and strictly prohibited. If you have received this electronis transmission in error, please immediately notify the sender by reply and permanently delete all copies of this message and its attachments. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dave Johnson Sent: Thursday, August 25, 2005 1:54 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Please help us name our new product Mercedes Medical is coming out with a new teflon coated microtome blade, but we need a name! We have 3 names up for consideration: 1. Advant-Edge 2. Cutting Edge 3. Platinum Blade. Let us know which you like, or come up with a new one and if we use it we'll send you a great gift. Also, come see us at our island booth at NSH in Ft. Lauderdale booth #229. Enter to win a 26" Flat screen TV, and pick up your free Henry the Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be held at the Annual Symposium/Convention from September 10-14 in Ft Lauderdale. You can register on line at www.NSH.org. Visit us on line at www.MercedesMedical.com and get 10% off your first order. Or call at 800.331.2716. _______________________________________________ Histonet mailing list Histonet@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 ---------------------------------------------------------- Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est exclusivement destin? au(x) destinataire(s), personnes physiques ou morales, qu'il d?signe. Il constitue de ce fait une correspondance ? caract?re priv? et peut contenir des informations confidentielles. Si ce message vous est parvenu par erreur, nous vous remercions d'en aviser imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de le d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver de copie. This message, including any attachment, is intended for the use of the individual or entity to which it is addressed. It is therefore to be considered as a private correspondence which may contain confidential information. If you are not the intended recipient, please advise the sender immediately by reply e.mail and delete this message and any attachment thereto without retaining a copy. ---------------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 DCadoretteHall <@t> sprg.mercy.net Fri Aug 26 10:03:53 2005 From: DCadoretteHall <@t> sprg.mercy.net (Cadorette-Hall, Diane M) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] RE: RE: Salary Survey Message-ID: <99648DDDA90EA647B3E8A1FD4AFD339801A2D9BB@sprgexelroy.sprg.mercy.net> I am receiving more emails on the subject of the salaries acorss the US and how the actual pay scales are higher than reported in the salary survey conducted by the ASCP than I expected. Most also report that their Human Resources departments usually just call hospitals in the same state to compare salaries and give marignal, if any, raises when a request to evaluate salaries is made by department supervisors. I would encourage you to respond to the entire group of HistoNetters so that the managers I have been hearing from will have additional information to present to their employers. Gathering this type of information is crucial to an acurate analysis of market value. > Diane Cadorette-Hall, HT (ASCP) > Pathology Laboratory Supervisor > St. Johns Regional Health Center > 1235 East Cherokee > Springfield, Missouri 65804 > (417) 820-6492 > Fax: (417) 820-7790 > DCadoretteHall@sprg.mercy.net > > From petert <@t> serotec.co.uk Fri Aug 26 10:10:41 2005 From: petert <@t> serotec.co.uk (Peter Tree) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Procedure for Mallory's Trichrome Stain Message-ID: try this link, http://stainsfile.info/StainsFile/stain/conektv/tri_mallory.htm From timothy.macatee <@t> med.nyu.edu Fri Aug 26 10:14:40 2005 From: timothy.macatee <@t> med.nyu.edu (Timothy Macatee) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Please help us name our new product In-Reply-To: Message-ID: How about Rollz-Off. Tim On 8/25/05 2:54 PM, "Dave Johnson" wrote: > Mercedes Medical is coming out with a new teflon coated microtome blade, but > we need a name! We have 3 names up for consideration: > 1. Advant-Edge > 2. Cutting Edge > 3. Platinum Blade. > > Let us know which you like, or come up with a new one and if we use it we?ll > send you a great gift. > > Also, come see us at our island booth at NSH in Ft. Lauderdale booth #229. > > > Enter to win a 26? Flat screen TV, and pick up your free Henry the > Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be held > at the Annual Symposium/Convention from September 10-14 in Ft Lauderdale. > You can register on line at www.NSH.org. > > > Visit us on line at www.MercedesMedical.com and get 10% off your first > order. Or call at 800.331.2716. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Tim Macatee Research Histology Core New York University School of Medicine 550 First Ave. Department of Pathology. Medical Science Building - Room 521 New York, N.Y. 10016 (212) 263-3888 From brandonedwardmills <@t> gmail.com Fri Aug 26 13:10:52 2005 From: brandonedwardmills <@t> gmail.com (Brandon Edward Mills) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] independent histology lab Message-ID: <9204fc2705082611104f98d963@mail.gmail.com> im semi-new to the whole histo-world, and i just have a quick question... are there any completely independent histology labs? not independent pathology labs with a histology lab branch or a few techs working within, but a truly independent "tech" lab, whether small mom&pop or a larger chain of labs. i hear a lot about histotechs being underpaid and underappreciated by pathologists, and it seems to me that someone would have branched away from the pathologist and created an independent tech lab that could possibly service a few larger patho labs and hospitals, and take advantage of economies of scale by doing so. any input? From liz <@t> premierlab.com Fri Aug 26 13:50:29 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] independent histology lab In-Reply-To: <9204fc2705082611104f98d963@mail.gmail.com> Message-ID: <000001c5aa6f$087711c0$a7d48a80@AMY> Brandon We are a small contract lab that services Universities, Pharmaceutical and Biotech companies. When I hire individuals for a histology position I first of all require a BS or HT or HTL certification. I have OTJ trained multiple individuals through the years who have become HTL certified. My last tech was hired with a BS. By the time she left my business she was both HTL certified and passed her QIHC. I feel that I pay a competitive wage, with full benefits (medical, dental and retirement). Its my personal opinion that my business will only be a good as the individuals that I hire, so I want to hire the best, which means I need to pay a competitive salary with benefits, etc. I just hired a new histotech, he's HT certified and did use the ASCP salary survey to judge an appropriate wage. I ended up with a starting salary that was in the mid to high range on the scale. I'm not speaking for all small contract labs, just for myself. Overall starting your own histology business is both challenging and rewarding. Lots of hours and sometimes lots of stress, but I wouldn't change a thing. I love my job. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 Brandon Edward Mills Sent: Friday, August 26, 2005 11:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] independent histology lab im semi-new to the whole histo-world, and i just have a quick question... are there any completely independent histology labs? not independent pathology labs with a histology lab branch or a few techs working within, but a truly independent "tech" lab, whether small mom&pop or a larger chain of labs. i hear a lot about histotechs being underpaid and underappreciated by pathologists, and it seems to me that someone would have branched away from the pathologist and created an independent tech lab that could possibly service a few larger patho labs and hospitals, and take advantage of economies of scale by doing so. any input? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LINDA.MARGRAF <@t> childrens.com Fri Aug 26 13:54:00 2005 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Re: NSH meet and greet Message-ID: Here's a message from Becky she had problems getting on the list.... . -----Original Message----- From: Orr, Rebecca Sent: Wednesday, August 24, 2005 1:39 PM To: 'histonet-request@lists.utsouthwestern.edu' Subject: NSH meeting and greet WOW!!!!!!! What a great response! I got a TON of yes replies! I'm in contact with some of you to see about a central meeting place.. There has been mention of a booth but as I see it, then we'll all saunter over to the booth on our own good time instead of all concentrating at one place and one time... So we'll set up the "OUTPOST" to be at the specific time of 6pm Monday Septemer 12. LOCATION WILL BE SENT TO YOU ALL VIA HISTONET VERY SOON. Thanks for your replies Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 From dholmes <@t> anatomy.umsmed.edu Fri Aug 26 15:41:22 2005 From: dholmes <@t> anatomy.umsmed.edu (Dianne Holmes) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Sihler's stain Message-ID: I have been asked to use the Sihler technique to stain nerves in the trachea. I have only 'heard' of this method and would like some input from someone who has actually used it. The reprint I was given to refer to is a 2001 experiment on rabbit eyes and it took up to 3 months to process the tissue !! Any help would be appreciated. From maria <@t> ski.org Fri Aug 26 17:34:22 2005 From: maria <@t> ski.org (Maria Mejia) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Sihler's stain References: Message-ID: <430F98EE.4040805@ski.org> Hello Dianne, I have not only heard of this stain, I have done it on thick whole monkey extra ocular muscle thick sections. The staining procedure is a simple one, however, don't be fooled - this is a challenging stain to do!!! The reference I have is titled "The Sihler stain: a technique for studying peripheral nerves in whole-mount specimens" by Bei-lian Wu & Ira Sanders. Over the course of several trials, I have modified the methodolgy from the article to fit what I needed. The following technical times are for whole thick monkey extra cellular muscle samples: **The total time required to stain varies & depends on the size & type of the tissue sample - so testing should be done (before) doing on the real McCoy samples. ***Each step (exception is the running water step) - should use agitation!!! Fixation: Ideally, tissue should be perfused w/10% UNNEUTRALIZED formalin/DH20 prior to removing the tissue. Immersion fixation technique of fresh tissue should also work. The slight amount of formic acid in formalin is less harmful in its effects upon nerve tissue than a neutralized solution. Remember to change the formalin fixing solution once a week - it takes 1 month or longer for larger whole- mount tissue. However, for my thick tissue sample, I fixed for only a few days to one (1) week (w/agitation). 1. place tissue samples in tissue cassettes & in a container wash in gentle running water - 30 minutes. 2. remove samples from cassettes & place one tissue sample in each large clear plastic well containing 3% potassium hydroxide w/0.2ml of hydrogen peroxide in 100ml DH20. the clear plastic box w/lid has 15 to 25 wells -depends on the size you buy. each well is clearly ID. **change this solution every other day & total time for tissue in this solution varies from 4-6 days. 3. specimen initially appears highly refractive, but will progressively lose this property. check specimen under a dissecting microscope every day until the soft tissue is faded, transparent & barely refractive while the nerves & especially the small nerve brances can be seen clearly as white fibers. **KHOH affects nerve tissue to a certain extent, so this step should be as brief as possible. the durration of this step varies w/each specimen size. may take 2 to 3 weeks for different wholemounts 4. remove tissue from KHOH solution, place tissue again in cassettes & wash in gentle H20 - 30 minutes. Staining of Tissue: Use the Sihler II solution to stain the specimen. 5. after washing, remove samples from cassettes & again place in plastic wells that contain working Ehrlich's hematoxylin. **this hematoxylin solution is very important!!! it must be at least 3 weeks old and NOT older that 6 months for this stain to work!!! I buy my Ehrlich's hem from Electron Microscopy Sciences 215-646-1566. ask to speak w/the technical person who makes the hematoxylin tell him what you need and to write the date on the bottle when the hematoxylin was made, so you can keep track of its usage peak!!! the time is important because it tends to under stain if not ripe enough & over stain if too old & therefore more difficult to remove excess stain from tissue during the de-stain step. **solution should be changed every other day & agitate. total time for staining should be about 4 to 5 days. **check tissue every day under dissecting scope. could take 2 to 3 wks if staining different thick wholemount tissue - don't know times for thinner sections. The endpoint is when all nerves including the finest brances are stained dark violet blue. for small or this specimens the amount of hematoxyling maybe decrease to as little as 0.5 parts. De-staining: use the Sihler I (working 0.5% acid solution) 6. remove tissue from wells & place in tissue cassettes...again - & wash in gentle running H20 - 10 minutes. 7. after washing, return samples into wells w/Sihler I acid solution - check tissue every 20 minutes & change solution if it turns blue!!! total time could take up to 2.5 hours - so check, check tissue under scope!!! ***check tissue by trans illuminating them while observing under the dissecting scope. end-point is when tissue has a transparent lavendar colored to clear soft tissue w/dark violet nerves. **if the nerve brances look faded or take on a yellow tint, the tissue is either over de-stained or there is too much acetic aicd in the Sihler sol. if this occurs, tissue must be washed thoroughly w/tap H20 & re-stained in Sihler solution II & then repeat the de-stain step again - this time use less glacial acetic acid. **but I found that my modified method was suitable for smaller, thinner or lightly stained tissues samples. Neurtralization: 8. after de-staining the tissue is acidic & needs to be neutralized. place sample in tissue cassettes...again, wash tissue thoroughly in running - 1 to 1.5 hours. 9. remove sample from cassetts & return to wells with freshly made 0.05% lithium carbonate (agitate). **check every 15 minutes & change solution if it turn pinkish color. leave samples in this solution until the black-blue fibers colored nerves turn to violet-blue - about 1.5 hours Clearing: 10. remove samples from wells & place in cassettes again to wash in running H20 - for 30 minutes to 1 hr. (I did 30 minutes). 11. remove samples from cassettes and place in several wells that contain increasingly graded glycerin (buy a good brand grade of glycerin) - start with: >40% glycerin/DH20 - overnight. >60% glycerin/DH20 - overnight. >80% glycerin/DH20 - 1 to 2 nights >99% glycerin - 2 to 3 nights and finally 100% glycerin - 4 nights. ** can store tissue in this last step, just add a few grains of thymol. change glycerin every 4 to 6 months using fresh 100% glycerin w/thymol. WOW, that's it!!! Yours Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria @ski.org Dianne Holmes wrote: >I have been asked to use the Sihler technique to stain nerves in the trachea. I have only 'heard' of this method and would like some input from someone who has actually used it. The reprint I was given to refer to is a 2001 experiment on rabbit eyes and it took up to 3 months to process the tissue !! Any help would be appreciated. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From jcolclefa <@t> aol.com Fri Aug 26 18:37:26 2005 From: jcolclefa <@t> aol.com (John PJ Coleman) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Mallory In-Reply-To: <200508261300.481430f4aba340@rly-xh01.mx.aol.com> References: <200508261300.481430f4aba340@rly-xh01.mx.aol.com> Message-ID: Mallory'sTrich- exactly what does your researcher want to see? On Aug 26, 2005, at 1:01 PM, histonet-request@lists.utsouthwestern.edu wrote: > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. Re: Please help us name our new product > (Stephen.Eyres@sanofi-aventis.com) > 2. Procedure for Mallory's Trichrome Stain (Snider, Deanna) > 3. Question about Re: [Histonet] EDTA recipe with PVP (Gayle Callis) > 4. RE: [BULK] Re: [Histonet] Please help us name our new > product (Barnhart, Tammy) > 5. RE: RE: Salary Survey (Cadorette-Hall, Diane M) > 6. RE: Procedure for Mallory's Trichrome Stain (Peter Tree) > 7. Re: Please help us name our new product (Timothy Macatee) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Fri, 26 Aug 2005 15:08:04 +0100 > From: Stephen.Eyres@sanofi-aventis.com > Subject: Re: [Histonet] Please help us name our new product > To: "Dana Marshall" > Cc: Histonet@lists.utsouthwestern.edu, Terry Costello > , histonet-bounces@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset=ISO-8859-1 > > > > > > Now look here chaps and chapesses, this has gone on long enough. I > don't > want to put a BLOCK on this, nor have I a SCORE to settle, and I don't > want > to WAX lyrical about blades. However, a good name will give you an EDGE > over the competition, who, after all, are always trying to STEEL a > march on > everyone else. A catchy name will make you a CUT above the rest, and in > sales terms BEVALuable. On the other hand, ours is only a small > SECTION of > the market with customers often FLOATING between brands after SOAKING > up > all the sales blurb. I think there is a need for TRIMMINGIN the blade > market, or so a promotional CASSETTE, wrapped in a nice RIBBON, > informed > me. In fact there were several cassettes, a SERIAL promotion I suppose > you > could say. I have lost one of them. Someone must have NICKED it. What > LEVELS some folk will stoop to. I think I've RAKEed up enough dust now > and > if I HEIFFOR see any more messages on this subject I could get into a > STROP > . I've had enough now. Time to go an HONE some of my other skills. Do > you > think I've gone off my ROCKER, CAMBRIDGE of course. What if my > COVERSLIPS? > Do you think someone will LABEL me a nutter? I'm getting tired now. In > fact > I'm CHATTERED. Wish someone would close the VENETIAN BLIND. Think I > need a > pint, my throats really DRY. > > Cheers > > Steve > > > > "Dana Marshall" > To: > "Terry Costello" > Sent by: "Dave > Johnson" > histonet-bounces@lists.utsouth > > western.edu cc: > Subject: > Re: [Histonet] Please help us name our new product > > 26/08/2005 14:36 > > > > > > i like this one! > dm > ----- Original Message ----- > From: "Terry Costello" > To: "Dave Johnson" ; > > Sent: Thursday, August 25, 2005 3:16 PM > Subject: RE: [Histonet] Please help us name our new product > > > "MERCEDES BLADZ" > > Terrence Costello > GTx, Inc. > Three North Dunlap > Third Floor > Van Vleet, Bldg. > Memphis, TN 38163 > (901) 523-9700 ext. 102 (office) > (901) 523-9772 (fax) > tcostello@gtxinc.com > > > _______________________________________________________________________ > _____________________________________________ > > > This electronic message, including any attachments, is confidential and > proprietary and is soley for the intended recipient. If you are not the > intended recipient, this message was sent to you in error and you are > hereby > advised that any review, disclosure, copying, distribution or use of > this > message, or any of the information included therein, is unauthorized > and > strictly prohibited. If you have received this electronis transmission > in > error, please immediately notify the sender by reply and permanently > delete > > all copies of this message and its attachments. > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dave > Johnson > Sent: Thursday, August 25, 2005 1:54 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Please help us name our new product > > Mercedes Medical is coming out with a new teflon coated microtome > blade, > but > we need a name! We have 3 names up for consideration: > 1. Advant-Edge > 2. Cutting Edge > 3. Platinum Blade. > > Let us know which you like, or come up with a new one and if we use it > we'll > send you a great gift. > > Also, come see us at our island booth at NSH in Ft. Lauderdale booth > #229. > > > Enter to win a 26" Flat screen TV, and pick up your free Henry the > Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be > held > at the Annual Symposium/Convention from September 10-14 in Ft > Lauderdale. > You can register on line at www.NSH.org. > > > Visit us on line at www.MercedesMedical.com and get 10% off your first > order. Or call at 800.331.2716. > > > > _______________________________________________ > Histonet mailing list > Histonet@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 > > > > > ---------------------------------------------------------- > Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est > exclusivement destin? au(x) destinataire(s), personnes physiques ou > morales, qu'il d?signe. > Il constitue de ce fait une correspondance ? caract?re priv? et peut > contenir des informations confidentielles. > Si ce message vous est parvenu par erreur, nous vous remercions d'en > aviser > imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de > le > d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver > de > copie. > > > This message, including any attachment, is intended for the use of the > individual or entity to which it is addressed. > It is therefore to be considered as a private correspondence which may > contain confidential information. > If you are not the intended recipient, please advise the sender > immediately > by reply e.mail and delete this message and any attachment thereto > without > retaining a copy. > ---------------------------------------------------------- > > > > > ------------------------------ > > Message: 2 > Date: Fri, 26 Aug 2005 10:46:04 -0400 > From: "Snider, Deanna" > Subject: [Histonet] Procedure for Mallory's Trichrome Stain > To: histonet@lists.utsouthwestern.edu > Message-ID: > <84BE46B37B314D409C5A17B7BAB022D616031A@IDC-EX-VS01.shriners.cc> > Content-Type: text/plain > > Hello again netters.... > I am trying to find a procedure for Mallory's Trichrome and am not > having an > easy time of it! Once was emailed to me, but it skips an entire > step.(one > of the reagents in's used at all!) I have done this stain in the > past, but > its been a while and I cannot remember the procedure at all. Please > help! > It can be emailed to me or faxed...513-872-6072. The Masson is not > demonstarting what the researcher wants to see. > Thanks in advance... > Deanna Snider HT ASCP > Shriners Hospital for Children > Research Dept. > Cincinnati, Oh > 513-872-6388 > > CONFIDENTIALITY NOTICE: This e-mail communication and any attachments > may > contain confidential and privileged information for the use of the > designated recipients. If you are not the intended recipient, (or > authorized to receive for the recipient) you are hereby notified that > you > have received this communication in error and that any review, > disclosure, > dissemination, distribution or copying of it or its contents is > prohibited. > If you have received this communication in error, please destroy all > copies > of this communication and any attachments and contact the sender by > reply > e-mail or telephone (813) 281-0300. > > > ------------------------------ > > Message: 3 > Date: Fri, 26 Aug 2005 09:00:49 -0600 > From: Gayle Callis > Subject: Question about Re: [Histonet] EDTA recipe with PVP > To: "Nancy W. Troiano" , > Histonet@lists.utsouthwestern.edu > Message-ID: > <6.0.0.22.1.20050826085927.01b7cf48@gemini.msu.montana.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Nancy, > > What is the advantage of adding PVP to the EDTA decalcifying > solution? Just curious. > > At 05:38 AM 8/26/2005, you wrote: >> Here is our recipe for EDTA containing PVP: Dissolve 41.3 grams EDTA >> (disodium salt), 4.4 grams Sodium Hydroxide and 50 g PVP-40 (Sigma) in >> 1000 ml distilled water over very low heat. Cool to room temperature, >> adjust pH to 7.4 once cooled. We use this to gently decalcify our >> mouse >> long bones, it will take about 3 weeks to decalcify an adult mouse >> femur. > > Gayle Callis > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 > 406 994-4303 (FAX) > > > > > > ------------------------------ > > Message: 4 > Date: Fri, 26 Aug 2005 10:01:42 -0500 > From: "Barnhart, Tammy" > Subject: RE: [BULK] Re: [Histonet] Please help us name our new > product > To: "'Stephen.Eyres@sanofi-aventis.com'" > , Dana Marshall > > Cc: Histonet@lists.utsouthwestern.edu, Terry Costello > , histonet-bounces@lists.utsouthwestern.edu > Message-ID: <1779904B5E82D511914C00D0B793339205BFD9EB@exchangent> > Content-Type: text/plain; charset="iso-8859-1" > > Bravo....Bravo.....lol > > -----Original Message----- > From: Stephen.Eyres@sanofi-aventis.com > [mailto:Stephen.Eyres@sanofi-aventis.com] > Sent: Friday, August 26, 2005 9:08 AM > To: Dana Marshall > Cc: Histonet@lists.utsouthwestern.edu; Terry Costello; > histonet-bounces@lists.utsouthwestern.edu > Subject: [BULK] Re: [Histonet] Please help us name our new product > Importance: Low > > > > > > > Now look here chaps and chapesses, this has gone on long enough. I > don't > want to put a BLOCK on this, nor have I a SCORE to settle, and I don't > want > to WAX lyrical about blades. However, a good name will give you an EDGE > over the competition, who, after all, are always trying to STEEL a > march on > everyone else. A catchy name will make you a CUT above the rest, and in > sales terms BEVALuable. On the other hand, ours is only a small > SECTION of > the market with customers often FLOATING between brands after SOAKING > up > all the sales blurb. I think there is a need for TRIMMINGIN the blade > market, or so a promotional CASSETTE, wrapped in a nice RIBBON, > informed > me. In fact there were several cassettes, a SERIAL promotion I suppose > you > could say. I have lost one of them. Someone must have NICKED it. What > LEVELS some folk will stoop to. I think I've RAKEed up enough dust now > and > if I HEIFFOR see any more messages on this subject I could get into a > STROP > . I've had enough now. Time to go an HONE some of my other skills. Do > you > think I've gone off my ROCKER, CAMBRIDGE of course. What if my > COVERSLIPS? > Do you think someone will LABEL me a nutter? I'm getting tired now. In > fact > I'm CHATTERED. Wish someone would close the VENETIAN BLIND. Think I > need a > pint, my throats really DRY. > > Cheers > > Steve > > > > > "Dana Marshall" > > To: > "Terry > Costello" > Sent by: "Dave > Johnson" > > histonet-bounces@lists.utsouth > > > western.edu cc: > > Subject: > Re: > [Histonet] Please help us name our new product > > > 26/08/2005 14:36 > > > > > > > > > i like this one! > dm > ----- Original Message ----- > From: "Terry Costello" > To: "Dave Johnson" ; > > Sent: Thursday, August 25, 2005 3:16 PM > Subject: RE: [Histonet] Please help us name our new product > > > "MERCEDES BLADZ" > > Terrence Costello > GTx, Inc. > Three North Dunlap > Third Floor > Van Vleet, Bldg. > Memphis, TN 38163 > (901) 523-9700 ext. 102 (office) > (901) 523-9772 (fax) > tcostello@gtxinc.com > > > _______________________________________________________________________ > _____ > ________________________________________ > > > This electronic message, including any attachments, is confidential and > proprietary and is soley for the intended recipient. If you are not the > intended recipient, this message was sent to you in error and you are > hereby > advised that any review, disclosure, copying, distribution or use of > this > message, or any of the information included therein, is unauthorized > and > strictly prohibited. If you have received this electronis transmission > in > error, please immediately notify the sender by reply and permanently > delete > > all copies of this message and its attachments. > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dave > Johnson > Sent: Thursday, August 25, 2005 1:54 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Please help us name our new product > > Mercedes Medical is coming out with a new teflon coated microtome > blade, > but > we need a name! We have 3 names up for consideration: > 1. Advant-Edge > 2. Cutting Edge > 3. Platinum Blade. > > Let us know which you like, or come up with a new one and if we use it > we'll > send you a great gift. > > Also, come see us at our island booth at NSH in Ft. Lauderdale booth > #229. > > > Enter to win a 26" Flat screen TV, and pick up your free Henry the > Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be > held > at the Annual Symposium/Convention from September 10-14 in Ft > Lauderdale. > You can register on line at www.NSH.org. > > > Visit us on line at www.MercedesMedical.com and get 10% off your first > order. Or call at 800.331.2716. > > > > _______________________________________________ > Histonet mailing list > Histonet@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 > > > > > ---------------------------------------------------------- > Le pr?sent message ainsi que ses ?ventuelles pi?ces jointes est > exclusivement destin? au(x) destinataire(s), personnes physiques ou > morales, qu'il d?signe. > Il constitue de ce fait une correspondance ? caract?re priv? et peut > contenir des informations confidentielles. > Si ce message vous est parvenu par erreur, nous vous remercions d'en > aviser > imm?diatement l'exp?diteur par retour de courrier ?lectronique puis de > le > d?truire, ainsi que ses ?ventuelles pi?ces jointes, sans en conserver > de > copie. > > > This message, including any attachment, is intended for the use of the > individual or entity to which it is addressed. > It is therefore to be considered as a private correspondence which may > contain confidential information. > If you are not the intended recipient, please advise the sender > immediately > by reply e.mail and delete this message and any attachment thereto > without > retaining a copy. > ---------------------------------------------------------- > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > 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: 5 > Date: Fri, 26 Aug 2005 10:03:53 -0500 > From: "Cadorette-Hall, Diane M" > Subject: [Histonet] RE: RE: Salary Survey > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > > <99648DDDA90EA647B3E8A1FD4AFD339801A2D9BB@sprgexelroy.sprg.mercy.net> > Content-Type: text/plain > > > I am receiving more emails on the subject of the salaries acorss the > US and > how the actual pay scales are higher than reported in the salary survey > conducted by the ASCP than I expected. Most also report that their > Human > Resources departments usually just call hospitals in the same state to > compare salaries and give marignal, if any, raises when a request to > evaluate salaries is made by department supervisors. I would > encourage you > to respond to the entire group of HistoNetters so that the managers I > have > been hearing from will have additional information to present to their > employers. Gathering this type of information is crucial to an acurate > analysis of market value. > > > >> Diane Cadorette-Hall, HT (ASCP) >> Pathology Laboratory Supervisor >> St. Johns Regional Health Center >> 1235 East Cherokee >> Springfield, Missouri 65804 >> (417) 820-6492 >> Fax: (417) 820-7790 >> DCadoretteHall@sprg.mercy.net >> >> > > > ------------------------------ > > Message: 6 > Date: Fri, 26 Aug 2005 16:10:41 +0100 > From: "Peter Tree" > Subject: RE: [Histonet] Procedure for Mallory's Trichrome Stain > To: "Snider, Deanna" > Cc: Histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset="iso-8859-1" > > try this link, > http://stainsfile.info/StainsFile/stain/conektv/tri_mallory.htm > > ------------------------------ > > Message: 7 > Date: Fri, 26 Aug 2005 11:14:40 -0400 > From: Timothy Macatee > Subject: Re: [Histonet] Please help us name our new product > To: Dave Johnson , > Histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset=UTF-8 > > How about Rollz-Off. > > Tim > > > On 8/25/05 2:54 PM, "Dave Johnson" wrote: > >> Mercedes Medical is coming out with a new teflon coated microtome >> blade, but >> we need a name! We have 3 names up for consideration: >> 1. Advant-Edge >> 2. Cutting Edge >> 3. Platinum Blade. >> >> Let us know which you like, or come up with a new one and if we use >> it we???ll >> send you a great gift. >> >> Also, come see us at our island booth at NSH in Ft. Lauderdale booth >> #229. >> >> >> Enter to win a 26??? Flat screen TV, and pick up your free Henry the >> Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be >> held >> at the Annual Symposium/Convention from September 10-14 in Ft >> Lauderdale. >> You can register on line at www.NSH.org. >> >> >> Visit us on line at www.MercedesMedical.com and get 10% off your first >> order. Or call at 800.331.2716. >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > -- > Tim Macatee > Research Histology Core > New York University School of Medicine > 550 First Ave. > Department of Pathology. > Medical Science Building - Room 521 > New York, N.Y. 10016 > (212) 263-3888 > > > > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 21, Issue 35 > **************************************** > JPJC-757 335-2159 http://journals.aol.com/jcolclefa/Waytoomuchboringcraptosayusually/ From MEeva <@t> mednet.ucla.edu Fri Aug 26 18:42:57 2005 From: MEeva <@t> mednet.ucla.edu (Eeva, Mervi) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Cox-2 on human colon Message-ID: <1304F6F98344D61190180002A51311C0085AAF96@medmail4.mednet.ucla.edu> Hi, Does anyone have suggestions about well working COX-2 antibody and protocol on human colon tissue immunohistochemistry ? We have got it working well on other tissues but colon is for some reason a problem: either too much background or too little staining. Thanks, - Mervi David Geffen School of Medicine at UCLA Gonda Center 695 Charles E. Young Drive South Los Angeles, CA 90095-7088 U.S.A. ---------------------------------------------------------- IMPORTANT WARNING: This email (and any attachments) is only intended for the use of the person or entity to which it is addressed, and may contain information that is privileged and confidential. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Unauthorized redisclosure or failure to maintain confidentiality may subject you to federal and state penalties. If you are not the intended recipient, please immediately notify us by return email, and delete this message from your computer. ---------------------------------------------------------- From WWmn916 <@t> aol.com Fri Aug 26 23:35:06 2005 From: WWmn916 <@t> aol.com (WWmn916@aol.com) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Dried-out muscle biopsy Message-ID: Hello everyone, I hoping someone can help me troubleshoot frozen muscle biopsies that are sometimes extremely difficult to cut because it's so dried out. The muscle looks like it would be easy to cut, but ends up being difficult and sections hard to get. What would "dry-out" muscle tissue? Every now and then, some muscle biopsies are so dried out that they seem like powder coming off the cutting blade. Three things come to mind as possible reasons: Specimen dried out shortly after procedure and transport, during freezing the muscle, or the cryostat is too cold. Thanks for thoughts and responses! Deb King Sacramento, CA From marjoh3 <@t> telus.net Sat Aug 27 12:22:26 2005 From: marjoh3 <@t> telus.net (Marilyn Johnson) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Newcastles Disease by Immunohistochemistry Message-ID: <002301c5ab2b$e55b5a80$6401a8c0@VALUED20606295> Hi Veterinary Histonetters, Hope there is someone who is staining avian tissues for Newcastles disease by IHC. Could you please provide me with information including the source of primary antisera, company, ordering information, phone/FAX no.and staining procedure, if possible? Hopefully, the staining procedure can be linked with Dako reagents (LSAB2 or Envision systems). Any replies would be greatly appreciated. Thankyou in advance. Marilyn Johnson Food Safety Division Alberta Agriculture Edmonton, Alberta, Canada From petepath <@t> yahoo.com Sat Aug 27 15:06:46 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] A techniqe for thin slicing very soft tissues Message-ID: <20050827200646.39475.qmail@web30415.mail.mud.yahoo.com> For anyone grossing out there ,I just put up a technique on my web site that I have used for thin slicing soft difficult to cut tissues. It requires nothing more than a good paper towel. It is great for breast biopsies recieved for frozen section. As far as I know it is an original technique. Let me know if anyone has ever used this before. I call it " The sausage technique". http://pathologyinnovations.com/surg_path_offerings.htm Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From AnthonyH <@t> chw.edu.au Sun Aug 28 18:14:10 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Levels and Recuts Message-ID: A great method of dealing with the problem A good example of scientific/pathologist communication that results in excellent quality. Terry, I wish I could be "inflammatory" (I calm down by mid-week) but I can't. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall Terry Dr,Consultant Histopathologist Sent: Friday, 26 August 2005 9:30 PM To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Levels and Recuts Another subject talked about several times before. I agree it is sensible to call another section (usually 'cos there is something technically wrong with the first), a recut. I usually scribble on the form, something like, "bloody rubbish" or "?Q.C.", or something similarly inflammatory:-) Levels are just a few extra sections at an arbitrary interval. Serial sections are self explanatory, and semi-serial is a combination of serial and levels - cut in a bit then do a burst of serials, then repeat. Sometimes I ask for levels through the block. What is asked for depends and what you are looking for and how you expect to find it. Invariably, for anything other than levels, I talk about the problem to the tech., so that they can cut with an eye to the problem. 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: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 25 August 2005 21:45 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Levels and Recuts In my training "levels" were " step sections" going a given number of turns of the wheel between each section. A recut was another slide made trimming as little as possible. I have heard people refer to recuts as I described "levels". I think this leads to confusion. I do not know what the official definitions are but if there is a vote, I like it the way I described. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From DDDeltour <@t> mar.med.navy.mil Mon Aug 29 04:11:27 2005 From: DDDeltour <@t> mar.med.navy.mil (DDDeltour@mar.med.navy.mil) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Please help us name our new product Message-ID: <080566D001A3D9459FFC0A391A646C9106C49F93@marxchg03.mar.med.navy.mil> How about Cut-Rite? -----Original Message----- From: Timothy Macatee [mailto:timothy.macatee@med.nyu.edu] Sent: Friday, August 26, 2005 11:15 AM To: Dave Johnson; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Please help us name our new product How about Rollz-Off. Tim On 8/25/05 2:54 PM, "Dave Johnson" wrote: > Mercedes Medical is coming out with a new teflon coated microtome blade, but > we need a name! We have 3 names up for consideration: > 1. Advant-Edge > 2. Cutting Edge > 3. Platinum Blade. > > Let us know which you like, or come up with a new one and if we use it we?ll > send you a great gift. > > Also, come see us at our island booth at NSH in Ft. Lauderdale booth #229. > > > Enter to win a 26? Flat screen TV, and pick up your free Henry the > Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be held > at the Annual Symposium/Convention from September 10-14 in Ft Lauderdale. > You can register on line at www.NSH.org. > > > Visit us on line at www.MercedesMedical.com and get 10% off your first > order. Or call at 800.331.2716. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Tim Macatee Research Histology Core New York University School of Medicine 550 First Ave. Department of Pathology. Medical Science Building - Room 521 New York, N.Y. 10016 (212) 263-3888 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Mon Aug 29 07:32:12 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Please help us name our new product Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB439F@fh2k093.fhmis.net> The Z stands for ....ZORRO!!!!! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: Histonet@lists.utsouthwestern.edu Sent: 8/26/2005 9:51 AM Subject: RE: [Histonet] Please help us name our new product I agree! Sarah > ---------- > From: Dana Marshall > Sent: Friday, August 26, 2005 9:36 AM > To: Terry Costello; Dave Johnson; Histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Please help us name our new product > > i like this one! > dm > ----- Original Message ----- > From: "Terry Costello" > To: "Dave Johnson" ; > > Sent: Thursday, August 25, 2005 3:16 PM > Subject: RE: [Histonet] Please help us name our new product > > > "MERCEDES BLADZ" > > Terrence Costello > GTx, Inc. > Three North Dunlap > Third Floor > Van Vleet, Bldg. > Memphis, TN 38163 > (901) 523-9700 ext. 102 (office) > (901) 523-9772 (fax) > tcostello@gtxinc.com > > > ________________________________________________________________________ __ > __________________________________________ > > This electronic message, including any attachments, is confidential and > proprietary and is soley for the intended recipient. If you are not the > intended recipient, this message was sent to you in error and you are > hereby > advised that any review, disclosure, copying, distribution or use of this > message, or any of the information included therein, is unauthorized and > strictly prohibited. If you have received this electronis transmission in > error, please immediately notify the sender by reply and permanently > delete > all copies of this message and its attachments. > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dave > Johnson > Sent: Thursday, August 25, 2005 1:54 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Please help us name our new product > > Mercedes Medical is coming out with a new teflon coated microtome blade, > but > we need a name! We have 3 names up for consideration: > 1. Advant-Edge > 2. Cutting Edge > 3. Platinum Blade. > > Let us know which you like, or come up with a new one and if we use it > we'll > send you a great gift. > > Also, come see us at our island booth at NSH in Ft. Lauderdale booth #229. > > > Enter to win a 26" Flat screen TV, and pick up your free Henry the > Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be held > at the Annual Symposium/Convention from September 10-14 in Ft Lauderdale. > You can register on line at www.NSH.org. > > > Visit us on line at www.MercedesMedical.com and get 10% off your first > order. Or call at 800.331.2716. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tlclab <@t> free.fr Mon Aug 29 09:59:23 2005 From: tlclab <@t> free.fr (TLCLab) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Please help us name our new product In-Reply-To: <07AB60D5D7B9754EBF56F360F98D083DEB439F@fh2k093.fhmis.net> References: <07AB60D5D7B9754EBF56F360F98D083DEB439F@fh2k093.fhmis.net> Message-ID: <431322CB.4070104@free.fr> What about Gold-edge or...Ramp'edge ?... Florent. Bonner, Janet a ?crit : > >The Z stands for ....ZORRO!!!!! > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >To: Histonet@lists.utsouthwestern.edu >Sent: 8/26/2005 9:51 AM >Subject: RE: [Histonet] Please help us name our new product > >I agree! >Sarah > > > >>---------- >>From: Dana Marshall >>Sent: Friday, August 26, 2005 9:36 AM >>To: Terry Costello; Dave Johnson; Histonet@lists.utsouthwestern.edu >>Subject: Re: [Histonet] Please help us name our new product >> >>i like this one! >>dm >>----- Original Message ----- >>From: "Terry Costello" >>To: "Dave Johnson" ; >> >>Sent: Thursday, August 25, 2005 3:16 PM >>Subject: RE: [Histonet] Please help us name our new product >> >> >>"MERCEDES BLADZ" >> >>Terrence Costello >>GTx, Inc. >>Three North Dunlap >>Third Floor >>Van Vleet, Bldg. >>Memphis, TN 38163 >>(901) 523-9700 ext. 102 (office) >>(901) 523-9772 (fax) >>tcostello@gtxinc.com >> >> >> >> >> >________________________________________________________________________ >__ > > >>__________________________________________ >> >>This electronic message, including any attachments, is confidential >> >> >and > > >>proprietary and is soley for the intended recipient. If you are not >> >> >the > > >>intended recipient, this message was sent to you in error and you are >>hereby >>advised that any review, disclosure, copying, distribution or use of >> >> >this > > >>message, or any of the information included therein, is unauthorized >> >> >and > > >>strictly prohibited. If you have received this electronis transmission >> >> >in > > >>error, please immediately notify the sender by reply and permanently >>delete >>all copies of this message and its attachments. >> >> >>-----Original Message----- >>From: histonet-bounces@lists.utsouthwestern.edu >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dave >>Johnson >>Sent: Thursday, August 25, 2005 1:54 PM >>To: Histonet@lists.utsouthwestern.edu >>Subject: [Histonet] Please help us name our new product >> >>Mercedes Medical is coming out with a new teflon coated microtome >> >> >blade, > > >>but >>we need a name! We have 3 names up for consideration: >>1. Advant-Edge >>2. Cutting Edge >>3. Platinum Blade. >> >>Let us know which you like, or come up with a new one and if we use it >>we'll >>send you a great gift. >> >>Also, come see us at our island booth at NSH in Ft. Lauderdale booth >> >> >#229. > > >>Enter to win a 26" Flat screen TV, and pick up your free Henry the >>Histo-Potamus T-shirt and new Histology Catalog. The NSH show will be >> >> >held > > >>at the Annual Symposium/Convention from September 10-14 in Ft >> >> >Lauderdale. > > >>You can register on line at www.NSH.org. >> >> >>Visit us on line at www.MercedesMedical.com and get 10% off your first >>order. Or call at 800.331.2716. >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From TMcNemar <@t> lmhealth.org Mon Aug 29 10:37:41 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Levels and Recuts Message-ID: <6CD94D97ED7D924BA5C2B588FA9528213967F7@mail.lmhealth.org> We used to have some confusion over these requests so I defined each and added it to the bottom of the request sheet. That way paths and techs would know exactly what they were requesting and what they were to cut. I defined them as follows: Recut: 1 additional slide, superficial cut Levels: multiple slides, step sections deeper into the block Serial sections: multple slides, every section on a slide Exhaust: multiple slides, multiple levels completely through tissue We generally cut 3 to 4 levels when requested. When we exhaust a block, the path generally wants only 6 levels. We throw out the rest. We rarely get a request for serial sections but we have a path that would order "Serial sections to exhaust". After a couple of times of receiving 50 or 60 slides, he quit that. For alot of our blocks, we cut levels at the time of the original H&E. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Patricia Karlisch [mailto:pkarlisch@psu.edu] Sent: Thursday, August 25, 2005 4:08 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Levels and Recuts Hello all, Can anyone reference a description for 'levels' vs 'recuts'. We use the terms to mean specific procedures in cutting. Thus, RECUTS are sections directly off the top of a previously cut block. Levels are 'recuts' but at different distances between sections. Generally, when we cut levels we toss the unwanted sections out. In some cases the unstained are kept. Does anyone in histology consider these to be 'routine' terms and is this defined in any textbook? I appreciate your input on this rather basic question. Pat Karlisch Supervisor, Histology, Pathology and Laboratory Medicine Penn State Milton S. Hershey Medical Center Mail Code H179 Hershey, PA 17033 Phone (717) 531-6072 Fax: (717) 531- 7741 email: pkarlisch@psu.edu *****E-Mail Confidentiality Notice***** This message (including any attachments) contains information intended for a specific individual(s) and purpose that may be privileged, confidential or otherwise protected from disclosure pursuant to applicable law. Any inappropriate use, distribution or copying of the message is strictly prohibited and may subject you to criminal or civil penalty. If you have received this transmission in error, please reply to the sender indicating this error and delete the transmission from your system immediately. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> comcast.net Mon Aug 29 11:02:28 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Levels and Recuts Message-ID: <082920051602.28960.431331940009932F000071202207001641CECE030E9D0C9A03@comcast.net> I understand the levels concept however we always set a specific number of sections between levels as 5 or 10 depending on the block size and thickness requested. That way we knew how deep we were in the block and of course exhaust the block is obvious. Is that not done anymore? Pam Marcum -------------- Original message -------------- > We used to have some confusion over these requests so I defined each and > added it to the bottom of the request sheet. That way paths and techs would > know exactly what they were requesting and what they were to cut. I defined > them as follows: > > Recut: 1 additional slide, superficial cut > Levels: multiple slides, step sections deeper into the block > Serial sections: multple slides, every section on a slide > Exhaust: multiple slides, multiple levels completely through tissue > > We generally cut 3 to 4 levels when requested. When we exhaust a block, the > path generally wants only 6 levels. We throw out the rest. We rarely get a > request for serial sections but we have a path that would order "Serial > sections to exhaust". After a couple of times of receiving 50 or 60 slides, > he quit that. > > For alot of our blocks, we cut levels at the time of the original H&E. > > Tom McNemar, HT(ASCP) > Histology Co-ordinator > Licking Memorial Hospital > Newark, Ohio 43055 > > > > -----Original Message----- > From: Patricia Karlisch [mailto:pkarlisch@psu.edu] > Sent: Thursday, August 25, 2005 4:08 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Levels and Recuts > > > Hello all, > Can anyone reference a description for 'levels' vs 'recuts'. We use > the terms to mean specific procedures in cutting. Thus, RECUTS are > sections directly off the top of a previously cut block. Levels are > 'recuts' but at different distances between sections. Generally, when > we cut levels we toss the unwanted sections out. In some cases the > unstained are kept. Does anyone in histology consider these to be > 'routine' terms and is this defined in any textbook? > I appreciate your input on this rather basic question. > > > Pat Karlisch > Supervisor, Histology, Pathology and Laboratory Medicine > Penn State Milton S. Hershey Medical Center > Mail Code H179 > Hershey, PA 17033 > Phone (717) 531-6072 > Fax: (717) 531- 7741 > email: pkarlisch@psu.edu > > *****E-Mail Confidentiality Notice***** > This message (including any attachments) contains information intended > for a specific individual(s) and purpose that may be privileged, > confidential or otherwise protected from disclosure pursuant to > applicable law. Any inappropriate use, distribution or copying of the > message is strictly prohibited and may subject you to criminal or civil > penalty. If you have received this transmission in error, please reply > to the sender indicating this error and delete the transmission from > your system immediately. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From algranth <@t> u.arizona.edu Mon Aug 29 13:07:10 2005 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] frozens on eye parts Message-ID: <4.3.2.7.2.20050829110234.00ccf338@algranth.inbox.email.arizona.edu> Histonetters, There is an investigator here who will soon be working with human eye "parts" that are frozen and sectioned on a cryostat. Right now they are not certain of what "parts" other than macula. Can these be snap frozen they way we do livers and kidneys, etc. or is there something special about eye tissue that requires some alternative freezing procedure to produce a better section? Thanks. Andi Grantham ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From doninn <@t> mail.nih.gov Mon Aug 29 15:32:09 2005 From: doninn <@t> mail.nih.gov (Donin, Nick (NIH/NCI)) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Fbxw7/hCDC4 Message-ID: <16A0583FB1644E4DB8C0A0265028B6FD02B4E7DA@nihexchange13.nih.gov> Histonetters, I am looking for an antibody for human Fbxw7 aka hCDC4 that works in FFPE mouse brain. I already have the Abcam antibody that works in the western blot, but was wondering if anyone has tried in paraffin. Any help in this is greatly appreciated. Thanks so much, Nick Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 From liz <@t> premierlab.com Mon Aug 29 15:37:41 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] trypsin verses chymotrypsin Message-ID: <000001c5acd9$808ef3a0$a7d48a80@AMY> Hello everyone Is there anyone out there that can explain the difference between trypsin and chymotrypsin for retrieval in immunohistochemistry. I'm working out a protocol for aggrecan on cartilage and I have a protocol that says they retrieved in chymotrypsin. Any input would be appreciated. Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 pruegg <@t> ihctech.net Mon Aug 29 16:52:09 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] independent histology lab In-Reply-To: <000001c5aa6f$087711c0$a7d48a80@AMY> Message-ID: <200508292152.j7TLq1nQ013836@chip.viawest.net> Brandon, I too have my own independent Histopathology Services lab. I worked in pathology at the University for 25 years and retired to become a master gardener. Well, this lab and all the equipment I would need to open it fell into my lap and I postponed the plans to become a master gardener. Because of my association with the University for so many years I have lots of contacts from there who send me work, I also do at times pay my Pathologist colleagues for a consult for my customers. I am also very involved in the histology world thru NSH and ASCP, so I get a lot of referrals from my histology friends. I am now doing work for investigators from all over the USA, I even do some international work. My business is booming. I am not as big or established as Liz but I have more than enough work to keep me going. I have a University student who works with me part-time, (full time in the summer), she is a Jr. biology major, I am training her to be a histotech, she does want to take the HTL certification exam. I am helping to pay her school tuition besides the hourly wage I pay here. I work very hard and get paid pretty well. Like Liz, I love what I do, I get a new project about every day. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Friday, August 26, 2005 11:50 AM To: 'Brandon Edward Mills'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] independent histology lab Brandon We are a small contract lab that services Universities, Pharmaceutical and Biotech companies. When I hire individuals for a histology position I first of all require a BS or HT or HTL certification. I have OTJ trained multiple individuals through the years who have become HTL certified. My last tech was hired with a BS. By the time she left my business she was both HTL certified and passed her QIHC. I feel that I pay a competitive wage, with full benefits (medical, dental and retirement). Its my personal opinion that my business will only be a good as the individuals that I hire, so I want to hire the best, which means I need to pay a competitive salary with benefits, etc. I just hired a new histotech, he's HT certified and did use the ASCP salary survey to judge an appropriate wage. I ended up with a starting salary that was in the mid to high range on the scale. I'm not speaking for all small contract labs, just for myself. Overall starting your own histology business is both challenging and rewarding. Lots of hours and sometimes lots of stress, but I wouldn't change a thing. I love my job. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 Brandon Edward Mills Sent: Friday, August 26, 2005 11:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] independent histology lab im semi-new to the whole histo-world, and i just have a quick question... are there any completely independent histology labs? not independent pathology labs with a histology lab branch or a few techs working within, but a truly independent "tech" lab, whether small mom&pop or a larger chain of labs. i hear a lot about histotechs being underpaid and underappreciated by pathologists, and it seems to me that someone would have branched away from the pathologist and created an independent tech lab that could possibly service a few larger patho labs and hospitals, and take advantage of economies of scale by doing so. any input? _______________________________________________ Histonet mailing list Histonet@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 tgoodpas <@t> fhcrc.org Mon Aug 29 17:26:16 2005 From: tgoodpas <@t> fhcrc.org (Goodpaster, Tracy A) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] trypsin verses chymotrypsin Message-ID: <8BD501B158A381409FE54DB421F82FCA012BB356@groucho.fhcrc.org> I would be interested in any comments on this topic as well! ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Elizabeth Chlipala Sent: Mon 8/29/2005 1:37 PM To: 'Histonet' Subject: [Histonet] trypsin verses chymotrypsin Hello everyone Is there anyone out there that can explain the difference between trypsin and chymotrypsin for retrieval in immunohistochemistry. I'm working out a protocol for aggrecan on cartilage and I have a protocol that says they retrieved in chymotrypsin. Any input would be appreciated. Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 tpmorken <@t> labvision.com Mon Aug 29 18:44:26 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] trypsin verses chymotrypsin Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D08C@usca0082k08.labvision.apogent.com> Liz, various enzymes break peptides at different sites. Maybe the authors found that chymotrypsin works better for cartlilage. See info at: http://www.greatvistachemicals.com/biochemicals/chymotrypsin.html 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 Elizabeth Chlipala Sent: Monday, August 29, 2005 1:38 PM To: 'Histonet' Subject: [Histonet] trypsin verses chymotrypsin Hello everyone Is there anyone out there that can explain the difference between trypsin and chymotrypsin for retrieval in immunohistochemistry. I'm working out a protocol for aggrecan on cartilage and I have a protocol that says they retrieved in chymotrypsin. Any input would be appreciated. Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager 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 DMBCMP <@t> aol.com Mon Aug 29 19:21:54 2005 From: DMBCMP <@t> aol.com (DMBCMP@aol.com) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Are Paraffin Blocks Contaminated? Message-ID: <1a7.3dbd1a3b.304500a2@aol.com> Hello all. I am inquiring of the latest ruling by CAP (or whoever) as to whether paraffin blocks are considered contaminated or hazardous. Are the employees who package them up for mailing required to be trained in the handling of possible exposure? .....as CJ brain blocks? Anyone know if this is necessary? Thanks very much. Dannie Blake, HT Sierra Pathology Lab Fresno, California From katri <@t> cogeco.ca Mon Aug 29 21:00:59 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Egg white/skim milk for biotin block Message-ID: <006e01c5ad06$aadfa640$6a9a9618@Katri> Hi Histonetters, I got this recipe created by Dr Miller, from Histonet archives. Egg white in distilled water is the source of avidin and skim milk source of biotin. It worked well for me until I bought skim milk from a different dairy producer and this milk does not seem to have any (enough) free biotin in it, so results were a disaster. Does anyone use skim milk powder for this purpose (to block endogenous biotin) and at what percentage? Katri Katri Tuomala Hamilton, Ontario, Canada From brucea <@t> unimelb.edu.au Tue Aug 30 00:50:07 2005 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] detection of lipopolysaccharide (LPS) Message-ID: Dear Histonetters, Am wondering whether any of you, know of/recommend a good staining technique for the detection of lipopolysaccharide (LPS). Silver staining is often done to detect H. pylori in situ, I think because it does stain LPS, but I am wondering whether I could use it to visualize an enriched flagella prep (the flagella are about 1-2 um long). Appreciate any response, Cheers - Bruce in OZ -- BRUCE ABALOZ PH:61383446282 HISTOLOGIST FAX:61383447909 DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au THE UNIVERSITY Of MELBOURNE. VICTORIA.AUSTRALIA 3010 http://www.zoology.unimelb.edu.au/ Nobody can make you feel inferior without your permission. - Eleanor Roosevelt DANCE LIKE NO-ONE'S WATCHING ******************************************************************************** This email and any attachments may contain personal information or information that is otherwise privileged, confidential or otherwise protected from disclosure/subject of copyright. Any use, disclosure or copying of any part of it is prohibited. The University does not warrant that this email or any attachments are free from viruses or defects. Please check any attachments for viruses and defects before opening them. If this email is received in error please delete it and notify us by return email. The University does not necessarily share or endorse the views expressed in this email. From brucea <@t> unimelb.edu.au Tue Aug 30 01:44:04 2005 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Omega Chemical's DECAL Message-ID: Dear Histonetters, Am wondering whether any of you, know of the product Omega Chemical's DECAL, which a PhD student in my lab. has informed me of. HAS a 24 hour 'turnaround' to fully decalcify a Mouse Head completely & doesn't interfere in Immuno. We have been using RDO successfully but even the potent RDO took 3 days to fully decalcify a mouse head & a 'Faster' turnaround' would be appreciated. Thankyou in advance for your advice, Bruce in OZ (on behalf of Nanette Schneider from Deutschland) -- BRUCE ABALOZ PH:61383446282 HISTOLOGIST FAX:61383447909 DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au THE UNIVERSITY Of MELBOURNE. VICTORIA.AUSTRALIA 3010 http://www.zoology.unimelb.edu.au/ Nobody can make you feel inferior without your permission. - Eleanor Roosevelt DANCE LIKE NO-ONE'S WATCHING ******************************************************************************** This email and any attachments may contain personal information or information that is otherwise privileged, confidential or otherwise protected from disclosure/subject of copyright. Any use, disclosure or copying of any part of it is prohibited. The University does not warrant that this email or any attachments are free from viruses or defects. Please check any attachments for viruses and defects before opening them. If this email is received in error please delete it and notify us by return email. The University does not necessarily share or endorse the views expressed in this email. From Kemlo.Rogerson <@t> elht.nhs.uk Tue Aug 30 02:26:31 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] independent histology lab Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD512@elht-exch1.xelht.nhs.uk> Interesting thought that appeals to the 'Techs' everywhere. Can a Histology Lab survive with no Medical Input? Probably can Vet Labs do. I suppose realistically the driver for a Lab comes the other way round; Medics finding Medics to service their needs; A completely 'Tech' driven Lab would probably not get the Contracts but that would belie that they would probably be more cost effective and almost certainly better run. It was explained to me once why Medics make poor Managers but for the life of me I can't remember, can anyone else? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Brandon Edward Mills Sent: 26 August 2005 19:11 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] independent histology lab im semi-new to the whole histo-world, and i just have a quick question... are there any completely independent histology labs? not independent pathology labs with a histology lab branch or a few techs working within, but a truly independent "tech" lab, whether small mom&pop or a larger chain of labs. i hear a lot about histotechs being underpaid and underappreciated by pathologists, and it seems to me that someone would have branched away from the pathologist and created an independent tech lab that could possibly service a few larger patho labs and hospitals, and take advantage of economies of scale by doing so. any input? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jorge.tornero <@t> gmail.com Tue Aug 30 05:03:51 2005 From: jorge.tornero <@t> gmail.com (Jorge Tornero) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Cutting defect and blade adjustement Message-ID: <8c964a7905083003033f3d4b7a@mail.gmail.com> Please help me about the right knife angle for cutting technovit 7100 blocks with a leica tungsten carbide knife. I use a leica RM 2145 microtome. I am using a freshly sharpened knife, but I get some sharp lines on the cuttings (you know, those that appear all the way long in the cut and in the same place) anyway. It is the first time that the knife is sharpened. Also I hear a buzzing noise when I cut, it seems like if the block causes some vibration in the blade, but it is well fastened. I have tried to decrease speed when cutting but it doesn't work at all. Now I am using a kind of moulds bigger than those that I used before, I don't know if I should trim the block edge to make it narrower and ease the cutting this way. any suggestions, please? Thank you very much Jorge Tornero IEO-CADIZ Spain From jorge.tornero <@t> gmail.com Tue Aug 30 06:05:19 2005 From: jorge.tornero <@t> gmail.com (Jorge Tornero) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Cutting defect and blade adjustement In-Reply-To: <1125398756.431438e41af0f@webmail.umdnj.edu> References: <8c964a7905083003033f3d4b7a@mail.gmail.com> <1125398756.431438e41af0f@webmail.umdnj.edu> Message-ID: <8c964a7905083004052844ed8@mail.gmail.com> My blocks are 12 mm wide by 16 mm long, and they are about 5 mm thick. They are glued (with technovit 3090) to a small piece of wood and this is the piece that I hold with the specimen holder. I am sectioning anchovy ovaries, and I am using a non standard protocol for the embedding, with much longer processing times, I guess because of the very high water content of the tissue. I wonder if it is better to make more changes in the alcohols instead of long times. I am sorry but no way for using glass knives or diamond (budgets) Thank you very much for your responses, I am waiting for more if you want, any help would be appreciated Jorge Tornero IEO-CADIZ Spain 2005/8/30, meyenhmf@umdnj.edu : > How big are your blocks? Thickness?Tissue? > I would use glass knives or Diamond. > Metal blades always give streaks. > Regards, > Markus > > > > forQuoting Jorge Tornero : > > > Please help me about the right knife angle for cutting technovit 7100 > > blocks with a leica tungsten carbide knife. I use a leica RM 2145 > > microtome. > > > > I am using a freshly sharpened knife, but I get some sharp lines on > > the cuttings (you know, those that appear all the way long in the cut > > and in the same place) anyway. It is the first time that the knife is > > sharpened. Also I hear a buzzing noise when I cut, it seems like if > > the block causes some vibration in the blade, but it is well fastened. > > I have tried to decrease speed when cutting but it doesn't work at > > all. Now I am using a kind of moulds bigger than those that I used > > before, I don't know if I should trim the block edge to make it > > narrower and ease the cutting this way. any suggestions, please? > > > > Thank you very much > > > > Jorge Tornero > > IEO-CADIZ > > Spain > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > -- > > From abright <@t> brightinstruments.com Tue Aug 30 06:15:48 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Cutting defect and blade adjustement Message-ID: Jorge, You do not say if these are vertical lines or horizontal. If they are vertical then you have a small nick in the knife, simply slide the knife to a new position. If the lines are horizontal then it could be a number of things. Please let me know. 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: Jorge Tornero [mailto:jorge.tornero@gmail.com] Sent: 30 August 2005 11:04 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cutting defect and blade adjustement Please help me about the right knife angle for cutting technovit 7100 blocks with a leica tungsten carbide knife. I use a leica RM 2145 microtome. I am using a freshly sharpened knife, but I get some sharp lines on the cuttings (you know, those that appear all the way long in the cut and in the same place) anyway. It is the first time that the knife is sharpened. Also I hear a buzzing noise when I cut, it seems like if the block causes some vibration in the blade, but it is well fastened. I have tried to decrease speed when cutting but it doesn't work at all. Now I am using a kind of moulds bigger than those that I used before, I don't know if I should trim the block edge to make it narrower and ease the cutting this way. any suggestions, please? Thank you very much Jorge Tornero IEO-CADIZ Spain _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jorge.tornero <@t> gmail.com Tue Aug 30 06:35:42 2005 From: jorge.tornero <@t> gmail.com (Jorge Tornero) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Cutting defect and blade adjustement In-Reply-To: References: Message-ID: <8c964a7905083004354171cd9a@mail.gmail.com> To tell you the truth, I have both. The worst are the vertical, because as you say, they seem to be caused by a tiny nick, but I have also tried with a brand new knif and it happen (less but it happen). I am a newby but the last year I cutted a lot of blocks and I was hoping to overcome those problems but even with brand new knifes it happen... About the horizontal stripes, (the venetian blind artifact or something like that) I guess it is caused by blocks that are too soft, but I will appreciate any help about it. I think maybe it is better to let the blocks dry or cure for a long time before cutting them. Here in Cadiz (SW Spain coast) it is very moist and warm and I think it may have lot of influence. Thank you for your help, I wait for more if you could ..... Jorge Tornero IEO-CADIZ Spain 2005/8/30, Alan Bright : > Jorge, > > You do not say if these are vertical lines or horizontal. If they are > vertical then you have a small nick in the knife, simply slide the knife > to a new position. If the lines are horizontal then it could be a number > of things. Please let me know. > > 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: Jorge Tornero [mailto:jorge.tornero@gmail.com] > Sent: 30 August 2005 11:04 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Cutting defect and blade adjustement > > > Please help me about the right knife angle for cutting technovit 7100 > blocks with a leica tungsten carbide knife. I use a leica RM 2145 > microtome. > > I am using a freshly sharpened knife, but I get some sharp lines on the > cuttings (you know, those that appear all the way long in the cut and in > the same place) anyway. It is the first time that the knife is > sharpened. Also I hear a buzzing noise when I cut, it seems like if the > block causes some vibration in the blade, but it is well fastened. I > have tried to decrease speed when cutting but it doesn't work at all. > Now I am using a kind of moulds bigger than those that I used before, I > don't know if I should trim the block edge to make it narrower and ease > the cutting this way. any suggestions, please? > > Thank you very much > > Jorge Tornero > IEO-CADIZ > Spain > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From mucram11 <@t> comcast.net Tue Aug 30 08:41:54 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Cutting defect and blade adjustement Message-ID: <083020051341.11279.43146221000C3B0500002C0F2200735446CECE030E9D0C9A03@comcast.net> Hi Jorge, Could you please give us your processing protocol? It would help to know if the schedule is too long and you over dehydrating if could cause you problems with very hard tissue components. If the blocks are not fully glued down and any wiggle at all ocurrs this could also be a problem. The product you are using contains GMA I believe and it is very water sensitive and will gather water from the just the enviroment so you may wish to stoe them in a chamber with a a dissicant to assure they are completely polymerized and free of excess water. Hope this helps. Pam Marcum UPenn Vet Schooll -------------- Original message -------------- > My blocks are 12 mm wide by 16 mm long, and they are about 5 mm thick. > They are glued (with technovit 3090) to a small piece of wood and this > is the piece that I hold with the specimen holder. I am sectioning > anchovy ovaries, and I am using a non standard protocol for the > embedding, with much longer processing times, I guess because of the > very high water content of the tissue. I wonder if it is better to > make more changes in the alcohols instead of long times. > > I am sorry but no way for using glass knives or diamond (budgets) > > Thank you very much for your responses, I am waiting for more if you > want, any help would be appreciated > > Jorge Tornero > IEO-CADIZ > Spain > > 2005/8/30, meyenhmf@umdnj.edu : > > How big are your blocks? Thickness?Tissue? > > I would use glass knives or Diamond. > > Metal blades always give streaks. > > Regards, > > Markus > > > > > > > > forQuoting Jorge Tornero : > > > > > Please help me about the right knife angle for cutting technovit 7100 > > > blocks with a leica tungsten carbide knife. I use a leica RM 2145 > > > microtome. > > > > > > I am using a freshly sharpened knife, but I get some sharp lines on > > > the cuttings (you know, those that appear all the way long in the cut > > > and in the same place) anyway. It is the first time that the knife is > > > sharpened. Also I hear a buzzing noise when I cut, it seems like if > > > the block causes some vibration in the blade, but it is well fastened. > > > I have tried to decrease speed when cutting but it doesn't work at > > > all. Now I am using a kind of moulds bigger than those that I used > > > before, I don't know if I should trim the block edge to make it > > > narrower and ease the cutting this way. any suggestions, please? > > > > > > Thank you very much > > > > > > Jorge Tornero > > > IEO-CADIZ > > > Spain > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@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 Sue.Kapoor <@t> uhsi.org Tue Aug 30 08:42:54 2005 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] microwave processing bloody specimens Message-ID: <61E9F2400F53D5119CFC00508B44E33B02E9E181@khmcexch.uhsi.org> Does anyone have a procedure for microwave processing bloody enodmetrial specimens that they would be willing to share?? Much thanks in advance! Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From mami_73 <@t> yahoo.com Tue Aug 30 08:54:07 2005 From: mami_73 <@t> yahoo.com (adrienn kiss) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] mouse heart epicardial fat Message-ID: <20050830135407.76859.qmail@web54109.mail.yahoo.com> Dear All, I am wondering whether anyone is experienced with gross and microscopic histology of mouse heart? Has anyone ever had the chance to detect epicardial fat depo in mouse hearts? How extensive is the fat in the epicardium? Is failing of the mouse heart linked to accumulation of fat with or without MI? Has anyone done histology in these or similar conditions? Paraffin sectioning/EtOH would withdraw the fat from the tissue. Thanks a lot, Dr. Adrienn Kis __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Ronnie_Houston <@t> bshsi.com Tue Aug 30 09:31:22 2005 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Supervisor Vacancy Message-ID: <530361BF03351B4CAE5270A05D3037B506B71A01@bsrexms01.bshsir.com> Bon Secours Richmond Health Systems, HealthPartners Laboratory has an immediate opening for a Technical Supervisor in our Histology department. This position requires an advanced level of knowledge acquired through a recognized Histology training program and a minimum of five (5) years experience in either a clinical or research histology laboratory. Certification of HT (ASCP) required; Bachelor of Science preferred. This person would report to the Anatomic Pathology Director and provides supervision for department bench activities, supervises the functioning of laboratory staff with work assignments, and analyzes workflow. Knowledge in IHC and FISH imperative; microwave processing a plus. Responsibilities would include research, troubleshooting and resolving related inquiries and problems-solving within the laboratory. We offer flexibility and an excellent benefits package ( http://bonsecours.com ). Qualified applicants please send/e-mail/fax resume with cover letter to me at the address below Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 2877972 (804) 2877906 - fax ronnie_houston@bshsi.com ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone, and permanently delete the original e-mail, attachment(s), and any copies. From dahmed <@t> mdanderson.org Tue Aug 30 09:47:19 2005 From: dahmed <@t> mdanderson.org (dahmed@mdanderson.org) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Part time and Full time positions with MD Anderson Cancer Center Message-ID: We currently have a part time and Full time technician vacancy in the Clinical Histology Lab. If you are interested, please contact Dianna Menard, tel:713-745-6184 or Deanna Petish, tel: 713-745-6110 in the Human Resource Department. Thank you. David S. Ahmed, HT(ASCP) Supervisor, Clinical Histology Laboratory Department of Pathology M.D. Anderson Cancer Center From MEeva <@t> mednet.ucla.edu Tue Aug 30 10:27:57 2005 From: MEeva <@t> mednet.ucla.edu (Eeva, Mervi) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Cox-2 on human colon Message-ID: <1304F6F98344D61190180002A51311C0085AAFA2@medmail4.mednet.ucla.edu> So far we have used Goat Polyclonal sc-1745 from Santa Cruz w/ Vector ABC kit and Rabbit Polyclonal Cat. No.160126 from Cayman Chemical w/ Envision kit. First one stains too much: background and all kind of cells. Later one doesn't give strong enough signal. Any experience of Transduction Labs antibody ? - Mervi -----Original Message----- From: Trajkovic, Dusko [mailto:dusko.trajkovic@pfizer.com] Sent: Tuesday, August 30, 2005 7:10 AM To: 'Eeva, Mervi' Subject: RE: [Histonet] Cox-2 on human colon Which antibody are you using, or which ones have you tried? Dusko -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Eeva, Mervi Sent: Friday, August 26, 2005 4:43 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Cox-2 on human colon Hi, Does anyone have suggestions about well working COX-2 antibody and protocol on human colon tissue immunohistochemistry ? We have got it working well on other tissues but colon is for some reason a problem: either too much background or too little staining. Thanks, - Mervi David Geffen School of Medicine at UCLA Gonda Center 695 Charles E. Young Drive South Los Angeles, CA 90095-7088 U.S.A. ---------------------------------------------------------- IMPORTANT WARNING: This email (and any attachments) is only intended for the use of the person or entity to which it is addressed, and may contain information that is privileged and confidential. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Unauthorized redisclosure or failure to maintain confidentiality may subject you to federal and state penalties. If you are not the intended recipient, please immediately notify us by return email, and delete this message from your computer. ---------------------------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. ---------------------------------------------------------- IMPORTANT WARNING: This email (and any attachments) is only intended for the use of the person or entity to which it is addressed, and may contain information that is privileged and confidential. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Unauthorized redisclosure or failure to maintain confidentiality may subject you to federal and state penalties. If you are not the intended recipient, please immediately notify us by return email, and delete this message from your computer. ---------------------------------------------------------- From jkiernan <@t> uwo.ca Tue Aug 30 10:32:16 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Omega Chemical's DECAL References: Message-ID: <43147C00.BB6B627B@uwo.ca> Should the claim, "... a 24 hour 'turnaround' to fully decalcify a Mouse Head completely & doesn't interfere in Immuno" be believed? Immuno for every antigen? If it decalcifies things rapidly it is almost certainly a quite concentrated acid, and may cause chemical changes such as Feulgen hydrolysis of DNA and extraction of RNA. Is the product's composition disclosed, or is it a secret brew? It's extremely easy to make decalcifying solutions in the lab from simple, inexpensive ingredients, following well tried and understood published recipes. -- ------------------------------- 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 _______________________________ Bruce Abaloz wrote: > > Dear Histonetters, > > Am wondering whether any of you, know of the product Omega Chemical's > DECAL, which a PhD student in my lab. has informed me of. HAS a 24 > hour 'turnaround' to fully decalcify a Mouse Head completely & > doesn't interfere in Immuno. > We have been using RDO successfully but even the potent RDO took 3 > days to fully decalcify a mouse head & a 'Faster' turnaround' would > be appreciated. Thankyou in advance for your advice, > > Bruce in OZ (on behalf of Nanette Schneider from Deutschland) > > -- > BRUCE ABALOZ PH:61383446282 > HISTOLOGIST FAX:61383447909 > DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au > THE UNIVERSITY Of MELBOURNE. > VICTORIA.AUSTRALIA 3010 > http://www.zoology.unimelb.edu.au/ From dahmed <@t> mdanderson.org Tue Aug 30 11:08:15 2005 From: dahmed <@t> mdanderson.org (dahmed@mdanderson.org) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Part time and Full time positions at MD Anderson Cancer Center- Addendum Message-ID: My apologies for neglecting to indicate location of facility. MD Anderson Cancer Center is located in Houston, Texas. David S. Ahmed, HT(ASCP) Supervisor, Clinical Histology Laboratory Department of Pathology M.D. Anderson Cancer Center From tpmorken <@t> labvision.com Tue Aug 30 11:09:10 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Tutorials available online Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D091@usca0082k08.labvision.apogent.com> I'd like to mention that the Lab Vision Online Tutorial program is going well with over 400 people signed up. We have 5 tutorials now: Basic Immunohistochemistry (1 CEU) Multiple Immunostaining (3 of 5 parts posted, 1 CEU for each part, authored by Chris van der Loos) Bioterrorism and the Histology Laboratory (1 CEU) The tutorials are free of charge and credits are given by the National Society for Histotechnology (NSH). They can be accessed at http://www.labvision.com/indexTutorial.cfm Tim Morken Lab Vision - Neomarkers From sjchtascp <@t> yahoo.com Tue Aug 30 11:41:25 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] glycerol cryoprotection Message-ID: <20050830164125.34327.qmail@web90207.mail.scd.yahoo.com> Has anyone used a 20% glycerol and 2%DMSO(in buffer or fixative) to cryoprotect tissue instead of 30% sucrose. Steve --------------------------------- Start your day with Yahoo! - make it your home page From cjohnson <@t> mail.rockefeller.edu Tue Aug 30 12:03:38 2005 From: cjohnson <@t> mail.rockefeller.edu (Carolyn Johnson) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] can PBS cause background? Message-ID: <200508301703.j7UH3e8c021004@smtp1.rockefeller.edu> Hello all, I have been struggling with high background in frozen mouse sections processed for immunofluorescence. A colleague told me that switching from PBS to a Tris buffer reduced their non-specific background. Does anyone know why this would make a difference? Also, I have tried using 1% sodium borohydride for my paraformaldehyde fixed sections, but the slide mounted tissue trapped gas bubbles and made it wrinkly. Is there a way to avoid the bubbles under the tissue? Thanks! Carolyn Johnson Research Assistant The Rockefeller University 1230 York Ave, Box 275 New York, NY 10021 Phone:(212) 327-8672 Fax:(212) 327-8664 cjohnson@rockefeller.edu From jcline <@t> wchsys.org Tue Aug 30 12:08:09 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Part time tech position Message-ID: <000001c5ad85$65f3a890$1d2a14ac@wchsys.org> I have a part time Histotech position. Forty hours per two week pay with one Saturday a month. Embedding, cutting staining, (accessioning on Sat) We average a little over 19,000 cases and about 81,000 slides per year. Location is Hagerstown, Maryland, about 70 miles outside of Baltimore or Washington, D.C. Located off a major interstate highway. ASCP registered or eligible. Please contact Mike Talhelm, personnel 301-665-4500 Joyce Cline, HT (ASCP) Technical Specialist, Pathology Hagerstown Medical Lab ***** 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 jkiernan <@t> uwo.ca Tue Aug 30 12:12:54 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] glycerol cryoprotection References: <20050830164125.34327.qmail@web90207.mail.scd.yahoo.com> Message-ID: <43149396.93FB8421@uwo.ca> This cryoprotectant mixture was introduced by Rosene, Roy & Davis (1986) J. Histochem. Cytochem. 24:1301-1315. They compared various cryoprotectants, including sucrose, in conjunction with various freezing techniques. Their conclusion was that the best results were given by 20% glycerol with 2% DMSO followed by freezing in isopentane at -75C (dry ice temperature). You can read the abstract at http://jhc.org/search.dtl Enter Rosene as an author, cryoprotection on the Title line, and click on Search. One item will then be shown. Click on the "Abstract" link. Below the abstract are links to about a dozen other papers (1996-2005) that cite the Rosene et al article. All are in high impact journals and full text .PDF files are available for aall of them. -- ------------------------------- 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 _______________________________ Steven Coakley wrote: > > Has anyone used a 20% glycerol and 2%DMSO(in buffer or fixative) to cryoprotect tissue instead of 30% sucrose. > > Steve > > > --------------------------------- > Start your day with Yahoo! - make it your home page > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From info <@t> instrumedics.com Tue Aug 30 13:01:29 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] frozens on eye parts References: <4.3.2.7.2.20050829110234.00ccf338@algranth.inbox.email.arizona.edu> Message-ID: <019601c5ad8c$f29d29e0$6401a8c0@INSTRUMEDICS22> Andrea, I believe that the eye should be snap frozen to minimize the growth of ice-crystals. We have prepared very good frozen eye sections with snap freezing followed by using the CryoJane Tape-Transfer process. To learn more about the tape-transfer technology please visit www.instrumedics.com. If you have any questions we would love to answer them. Bernice ----- Original Message ----- From: "Andrea Grantham" To: Sent: Monday, August 29, 2005 2:07 PM Subject: [Histonet] frozens on eye parts > Histonetters, > There is an investigator here who will soon be working with human eye > "parts" that are frozen and sectioned on a cryostat. Right now they are > not certain of what "parts" other than macula. Can these be snap frozen > they way we do livers and kidneys, etc. or is there something special > about eye tissue that requires some alternative freezing procedure to > produce a better section? > Thanks. > Andi Grantham > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > > > From VBlanc <@t> asterand.com Tue Aug 30 13:05:56 2005 From: VBlanc <@t> asterand.com (Vici Blanc) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Histotechnologist Position available in Detroit Message-ID: <56E63440ABF1FC4897947D5BA4CFA44F05A17B76@ATL1VEXC005.usdom004.tco.tc> All, Once again, Asterand is hiring! I welcome resumes for the following position: You may contact me at vblanc@asterand.com, or our HR manager, Ms. Christine Milne at cmilne@asterand.com Thank you! POSITION SUMMARY: Conducts research services on a custom basis for pharmaceutical and biotech client base. Performs complex laboratory procedures including but not limited to immunohistochemistry and in situ hybridization. Assists in the technical operation of a laboratory. Prepares histological sections for pathology review. Goals: * To assist in the development and management of an emerging revenue-generating department * To provide top-notch research services and data to drug discovery researchers * To become an integral member of a highly motivated and enthusiastic research team POSITION DUTIES AND RESPONSIBILTIES: * Design and perform simple and complicated scientific experiments based on customer requests * Make detailed observations and hypotheses * Perform immunohistochemistry, molecular testing, and in situ analysis on patient samples * Research and development of optimal experimental strategies to meet scientific needs of the customer * Data analysis and interpretation * Compile and prepare technical reports and experimental outcomes * Slide preparation for Pathology review * Section slides from formalin-fixed paraffin embedded and frozen/OCT embedded tissues * Cryosectioning, Microtome sectioning and tissue staining and cover slipping * Data entry and tracking of all products derived from sectioning * Work with laboratory automation * General laboratory and equipment maintenance * TMA sectioning and construction * Custom preparation of slides for sales orders * Other duties as assigned MINIMUM EXPERIENCE AND/OR EDUCATION REQUIRED: * Masters or Ph.D. in Biological or Medical science with emphasis on immunology, histology, cytology, molecular biology, biochemistry, or related field OR * Certified as a Histotechnologist by the ASCP or certified as a Histologic Technician by the ASCP and five years of experience in a clinical or research laboratory. * Equivalent education/experience will substitute for all minimum qualifications except when there are legal requirements such as license/certification/registration. PERSONAL ATTRIBUTES: * Excellent written and verbal communication skills * Interpersonal skills * Organizational skills and attention to detail * Problem solving skills * Creativity and latitude required * Rely on experience and judgment to plan and accomplish goals * Ability to multi-task PHYSCIAL DEMANDS: * May be required to stand for long hours WORKING CONDITIONS: * Occasionally required to work evenings, weekends * Pleasant working environment. RESPONSIBILITY FOR THE WORK OF OTHERS: * Work leadership responsibility: this job may assign day-to-day work to others; check their work; eliminate ordinary work difficulties they may encounter; and maintain their work flow; but does not have full supervisory responsibility. Victoria M. Blanc, Ph.D. Director of Experimental Pathology Services Asterand, Inc 440 Burroughs TechOne, suite 501 Detroit, MI 48202 Phone: 313-263-0960, ext. 240 Fax: 313-263-0961 www.asterand.com This e-mail message, together with any attachments, contains information belonging to Asterand, Inc. that may be confidential, proprietary, copyrighted and/or legally protected or privileged, and is intended for the sole use of the individual or entity named on this message. If you are not the intended recipient, and have received this e-mail message in error, please immediately return this message to its original sender and permanently delete it from your electronic and paper records. Thank You. From bamoe <@t> gundluth.org Tue Aug 30 13:29:07 2005 From: bamoe <@t> gundluth.org (bamoe@gundluth.org) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Full-time or part-time histotech position open - La Crosse Wisconsin Message-ID: We currently have a full-time or part-time position open in our busy clinical histology department at Gundersen Lutheran Medical Center in La Crosse Wisconsin. Full-time = 72 hours per 2-week pay period. Part-time = 48 hours (or flexible) per 2-week pay period. Day time hours, with on-call rotation one week every 6 - 7 weeks. Fun happy nutty bunch to work with! If interested please call soon! Thank you! Histology Lab direct dial - telephone 608-775-3139 - anyone would be glad to take your call! or Christine Kohlman - Dept. manager - telephone 608-775-5275 or visit the Gundersen Lutheran website at www.gundluth.org or see our ad on the NSH website at www.nsh.org under "employment" - go to Wisconsin Barb Moe Histology Department Gundersen Lutheran Medical Center La Crosse WI From sharon.osborn <@t> dnax.org Tue Aug 30 14:07:07 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] independent histo labs Message-ID: <29B25753F6B1D51196110002A589D44402398177@PALMSG30.us.schp.com> Brandon Mills, There are independent histology labs owned by enterprising histotechs throughout the country. You have Excalibur Labs in Moore, Okla to include in those who regularly participate in the histonet. Here in the Bay Area of California, I know of three labs that are histotech owned and operated for over 20 years. They are not regularly participants in the histonet so would not normally respond to your question. One is Histotech Laboratory in Hayward, CA. Another is located in Los Gatos, Ca and the other in the North Bay, somewhere around Sausalito. I have done business in the past with all three of these facilities. They employ several histotechs both in full time and part time capacities. The one in Hayward, CA does both animal and human tissues. All three run delivery/pick up services in this area for their customers. So, to answer your question, Yes, there are private facilities doing histology that are histotech run. The sizes range from one employee(owner) plus part time help to a number of techs. The labs here in the Bay Area (San Francisco) have from 10-25 employees working for them depending upon the work load. Are you interested in doing a survey to start your own practise? If so, there are a number of mentors who would probably be willing to provide feed back about how to make a more smooth business starting out due to their experiences. Sharon Osborn DNAX SP BioPharma 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 jstaruk <@t> masshistology.com Tue Aug 30 14:28:37 2005 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] independent histo labs In-Reply-To: <29B25753F6B1D51196110002A589D44402398177@PALMSG30.us.schp.com> Message-ID: <0IM100JJXVINMT00@vms044.mailsrvcs.net> Brandon, I started my company out of the basement of my home and have grown to include a new, state-of-the-art histopathology facility with many employees, pathologists and drivers. We do both Human, veterinary diagnostic and research/biotech work. I'd be happy to answer any questions you might have regarding starting your own histo business. Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Osborn, Sharon Sent: Tuesday, August 30, 2005 3:07 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] independent histo labs Brandon Mills, There are independent histology labs owned by enterprising histotechs throughout the country. You have Excalibur Labs in Moore, Okla to include in those who regularly participate in the histonet. Here in the Bay Area of California, I know of three labs that are histotech owned and operated for over 20 years. They are not regularly participants in the histonet so would not normally respond to your question. One is Histotech Laboratory in Hayward, CA. Another is located in Los Gatos, Ca and the other in the North Bay, somewhere around Sausalito. I have done business in the past with all three of these facilities. They employ several histotechs both in full time and part time capacities. The one in Hayward, CA does both animal and human tissues. All three run delivery/pick up services in this area for their customers. So, to answer your question, Yes, there are private facilities doing histology that are histotech run. The sizes range from one employee(owner) plus part time help to a number of techs. The labs here in the Bay Area (San Francisco) have from 10-25 employees working for them depending upon the work load. Are you interested in doing a survey to start your own practise? If so, there are a number of mentors who would probably be willing to provide feed back about how to make a more smooth business starting out due to their experiences. Sharon Osborn DNAX SP BioPharma 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eca9 <@t> georgetown.edu Tue Aug 30 14:59:18 2005 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] alpha-lactalbumin Message-ID: <4314BA96.80805@georgetown.edu> Hi, Does any more stain for alpha-lactalbumin? Could you please provide me with the name of your supplier and the staining protocol you use for this antibody? Thank you for all your help, Eva Andersson Georgetown University From fivetoads <@t> charter.net Tue Aug 30 22:08:17 2005 From: fivetoads <@t> charter.net (fivetoads@charter.net) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Eosin Leaching Message-ID: <48u2gp$1co7j10@mxip10a.cluster1.charter.net> We are having an intermittant but ongoing problem with our routing hematoxylin and eosin (H&E) stains leaching out the eosin after coverslipping anywhere from 1 hour to 1 day later. We use Bio-Care Cat-Hematoxylin and their eosin with phloxine, which is new to us this year. I do not think this brand has anything to do with the problem, as we have experienced this before. We use 4 absolute ethyl alcohol changes after the eosin for a total dehydration time of 6 minutes. After that we have 3 changes of Slide Brite (xylene substitute that we have used for the past 8 years)for a total clearing time of 6 minutes. This is all done on an automated stainer. We 'dump and rotate' the absolute alcohol solutions after every basket of 20-30 slides. We dump and rotate the clearing agent 2 times per week, more often when the slide count increases. (100 slides a day is a busy day for our small lab). We use a mounting medium that is designated for use with the clearing agent. All of! our ducks seem to be in 'a row'...what am I missing here?? Jonell Bay Area Hospital Coos Bay, Or From jorge.tornero <@t> gmail.com Wed Aug 31 00:47:49 2005 From: jorge.tornero <@t> gmail.com (Jorge Tornero) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Cutting defect and blade adjustement In-Reply-To: <6.0.0.22.1.20050830091624.01b64dd8@gemini.msu.montana.edu> References: <8c964a7905083003033f3d4b7a@mail.gmail.com> <6.0.0.22.1.20050830091624.01b64dd8@gemini.msu.montana.edu> Message-ID: <8c964a7905083022477598aa0@mail.gmail.com> Hello all, my knife is a leica TC knife, profile D. About the proccessing protocol, I'll give you the two protocols I've been using (last year and the protocol that we are trying now) Protocol A (Used last year) 2 Hours in 70% ethanol 2 Hours in 96% ethanol 1 hour in absolute ethanol 1 hour in a 1:1 mixture of base resin and 100% ethanol 48 hours in pure resin with hardener, at 4 degrees celsius Polimerization in oven at 37 degrees celsius, 24 hours. I have to ask you about this last step, because the instructions in heraeus-kulzer kits talk about 1 hour of polimerization at 23 ?C and subsequently another hour at 37 ?C, it sound confusing to me. Protocol B (To be used from now on) 32 hours in 70% ethanol 16 hours in 96% ethanol 8 hours in absolute ethanol 48 hours in 1:1 Resin:alcohol mixture 48 in pure resin+hardener Polimerization at room temperature Hope it helps you to help me, thank you very much Jorge Tornero IEO-CADIZ Spain 2005/8/30, Gayle Callis : > Wood is soft, and not as hard as the plastic - GMA, you should glue the > block with superglue to a metal chuck instead. > > Tungsten carbide knive will give lines in GMA, they are sharp but not as > sharp as glass. As long as you don't see defects in the tissue, ignore the > lines. Some people have great success with disposable blades, but be > careful where you buy the blades. Yes, I would trim trim down the block > edges to make it smaller, the less the plastic means less hardness you > have to go through. > > To adjust cutting angle, take a blank block without tissues to make angle > adjustments. Sharpening can change blades a bit, so you need to check that > angle again. > > You did not say whether the TC knife is a d profile or C profile, and you > can buy the latter. > > At 04:03 AM 8/30/2005, you wrote: > >Please help me about the right knife angle for cutting technovit 7100 > >blocks with a leica tungsten carbide knife. I use a leica RM 2145 > >microtome. > > > >I am using a freshly sharpened knife, but I get some sharp lines on > >the cuttings (you know, those that appear all the way long in the cut > >and in the same place) anyway. It is the first time that the knife is > >sharpened. Also I hear a buzzing noise when I cut, it seems like if > >the block causes some vibration in the blade, but it is well fastened. > >I have tried to decrease speed when cutting but it doesn't work at > >all. Now I am using a kind of moulds bigger than those that I used > >before, I don't know if I should trim the block edge to make it > >narrower and ease the cutting this way. any suggestions, please? > > > >Thank you very much > > > >Jorge Tornero > >IEO-CADIZ > >Spain > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 > 406 994-4303 (FAX) > > > From kcboswell <@t> grandecom.net Wed Aug 31 03:39:57 2005 From: kcboswell <@t> grandecom.net (Kevin and Chantel Bosewell) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Control Slides Message-ID: <000601c5ae07$94eef6e0$c9fb5a42@Rascal> Hello All, Our Pathologist believe control slides which are purchased are bomb proof and we should process our own. How does everyone else feel about this? Oh, and one more question, when you do any special stain do you review the patient slide or just the control? I have been asking around and I have been getting different answers. We recently had a false negative on a GMS for Fungus and this tech does not review the patient slides, in this case the control slide was positive. Thank you for any insight. Chantel Boswell HT (ASCP) Waco, Tx Thank you in advance, Chantel Boswell HT (ASCP) Waco, Tx From Kemlo.Rogerson <@t> elht.nhs.uk Wed Aug 31 04:06:59 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Control Slides Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD518@elht-exch1.xelht.nhs.uk> Just a thought but a control should be processed and fixed to the same standard as the Patient's material. I'd have thought logically that you can only control those factors yourself and that all controls should be 'home grown'. Except, that is, those that are unattainable due to their rarity. You should look at both the control and the patient's slide, if there is a disparity then you will have to restain; a known case of TB negative? Like any procedure things go wrong and as a Professional you ought to be in 'control'. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kevin and Chantel Bosewell Sent: 31 August 2005 09:40 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Control Slides Hello All, Our Pathologist believe control slides which are purchased are bomb proof and we should process our own. How does everyone else feel about this? Oh, and one more question, when you do any special stain do you review the patient slide or just the control? I have been asking around and I have been getting different answers. We recently had a false negative on a GMS for Fungus and this tech does not review the patient slides, in this case the control slide was positive. Thank you for any insight. Chantel Boswell HT (ASCP) Waco, Tx Thank you in advance, Chantel Boswell HT (ASCP) Waco, Tx _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Marirose.Satterfield <@t> MercyMemorial.org Wed Aug 31 05:32:10 2005 From: Marirose.Satterfield <@t> MercyMemorial.org (Satterfield, Marirose) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] sentinel biopsies Message-ID: <545CCCF1899E2042AA1C7A0DF101FC7C137356@mmh-01sv0101ew.mercymemorial.org> I would like to know how everyone is handling these biopsies. Extremely interested in if you facility does frozen sections or touch prints and what special handling (if any) you do. Our pathologists are discussing these options with a surgeon and we would like to know how everyone else is handling these specimens. I cannot find any CAP guidelines. Any input would be appreciated. M. Satterfield From BMolinari <@t> heart.thi.tmc.edu Wed Aug 31 05:49:48 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Gulf Coast Message-ID: My thoughts and prayers to all of you that are affected by Katrina. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) From ree3 <@t> leicester.ac.uk Wed Aug 31 06:08:45 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] RE: FERROPORTIN ANTIBODY Message-ID: -- I am after an antibody to ferroportin that works on rodent paraffin processed tissues..... Thanks Richard Edwards MRC TOXICOLOGY UNIT..U.K..... From Janet.Bonner <@t> FLHOSP.ORG Wed Aug 31 06:34:45 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] sentinel biopsies Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB43A5@fh2k093.fhmis.net> We receive the ndoes fresh, then do touch imprints - 4 slides, two for H&E and two for Diff-quik. Then process then routine Histology. For the Immuno part, the Pathologist may order a Sentinal Lymph Node Protocol: AE1, levels times three. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: 'histonet@pathology.swmed.edu' Sent: 8/31/2005 6:32 AM Subject: [Histonet] sentinel biopsies Importance: High I would like to know how everyone is handling these biopsies. Extremely interested in if you facility does frozen sections or touch prints and what special handling (if any) you do. Our pathologists are discussing these options with a surgeon and we would like to know how everyone else is handling these specimens. I cannot find any CAP guidelines. Any input would be appreciated. M. Satterfield _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Marirose.Satterfield <@t> MercyMemorial.org Wed Aug 31 06:59:55 2005 From: Marirose.Satterfield <@t> MercyMemorial.org (Satterfield, Marirose) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] sentinel biopsies Message-ID: <545CCCF1899E2042AA1C7A0DF101FC7C137357@mmh-01sv0101ew.mercymemorial.org> Do you concern yourself with the radiation issue? Isolate or process as routine? M. Satterfield From uince <@t> isbank.net.tr Wed Aug 31 07:20:55 2005 From: uince <@t> isbank.net.tr (Dr. Umit Ince) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] sentinel biopsies References: <545CCCF1899E2042AA1C7A0DF101FC7C137356@mmh-01sv0101ew.mercymemorial.org> Message-ID: <001801c5ae26$742ca3c0$bc00a8c0@umitince> We receive the nodes fresh. Bisection if the thickness is less than 0,6 cm. If more, we try to trim into three (or rarely four) slices. Than, prepare 2 slides from each surface. One imprint and one scrape, which are rapid H&E or MGG-quick stained. Than submit each slice in seperate cassettes. We do not concern about radiation issue and process as routine. We cut the the block without any trimming to avoid any tisue loss. Serial sections 50 microns apart are done, till the end of the tissue. This method usually produces a lot of slides. However one can be sure that, no micrometastasis is left behind in the block. If primary tumor diagnosis is lobular carcinoma, each serial section might be accompanied by one unstained positively charged slide for probable IHC. According to our experience (more than 21.000 surgicals and about 25.000 cytology cases annually) , frozen sectioning, does not give any additonal information compared to cytology slides and the risk of tissue loss or suboptimal morphology due to freezing artefacts is a real threat. U. Ince MD Pathologist ----- Original Message ----- From: "Satterfield, Marirose" To: Sent: Wednesday, August 31, 2005 1:32 PM Subject: [Histonet] sentinel biopsies >I would like to know how everyone is handling these biopsies. Extremely > interested in if you facility does frozen sections or touch prints and > what > special handling (if any) you do. Our pathologists are discussing these > options with a surgeon and we would like to know how everyone else is > handling these specimens. I cannot find any CAP guidelines. Any input > would > be appreciated. > M. Satterfield > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histolog <@t> fcv.unl.edu.ar Wed Aug 31 07:02:29 2005 From: histolog <@t> fcv.unl.edu.ar (Lab. Invest. Histol.) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Tutorials available online In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA20328D091@usca0082k08.labvision.apogent.com> Message-ID: You know other tutorials in PPT format??? Thanks!! -----Mensaje original----- De: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] En nombre de Morken, Tim - Labvision Enviado el: Martes, 30 de Agosto de 2005 13:09 Para: 'Histonet' Asunto: [Histonet] Tutorials available online I'd like to mention that the Lab Vision Online Tutorial program is going well with over 400 people signed up. We have 5 tutorials now: Basic Immunohistochemistry (1 CEU) Multiple Immunostaining (3 of 5 parts posted, 1 CEU for each part, authored by Chris van der Loos) Bioterrorism and the Histology Laboratory (1 CEU) The tutorials are free of charge and credits are given by the National Society for Histotechnology (NSH). They can be accessed at http://www.labvision.com/indexTutorial.cfm Tim Morken Lab Vision - Neomarkers _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Wed Aug 31 07:36:22 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] sentinel biopsies Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB43A6@fh2k093.fhmis.net> For the radiation specimens, we have a lead box. But for the most part the PA's do these ....good question, I'll check. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: 'histonet@pathology.swmed.edu' Sent: 8/31/2005 7:59 AM Subject: [Histonet] sentinel biopsies Do you concern yourself with the radiation issue? Isolate or process as routine? M. Satterfield _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Wed Aug 31 07:39:16 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] sentinel biopsies Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB43A7@fh2k093.fhmis.net> The PA's do use a HOT box but will perform the touch imprints first on bisected nodes. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: 'histonet@pathology.swmed.edu' Sent: 8/31/2005 7:59 AM Subject: [Histonet] sentinel biopsies Do you concern yourself with the radiation issue? Isolate or process as routine? M. Satterfield _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From japoteete <@t> saintfrancis.com Wed Aug 31 08:08:30 2005 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Control Slides Message-ID: We make all of our control slides, and I review both the patient slides and the control slides. I would rather catch a problem before I take the slides to the pathologist. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK -----Original Message----- From: Kevin and Chantel Bosewell [mailto:kcboswell@grandecom.net] Sent: Wednesday, August 31, 2005 3:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Control Slides Hello All, Our Pathologist believe control slides which are purchased are bomb proof and we should process our own. How does everyone else feel about this? Oh, and one more question, when you do any special stain do you review the patient slide or just the control? I have been asking around and I have been getting different answers. We recently had a false negative on a GMS for Fungus and this tech does not review the patient slides, in this case the control slide was positive. Thank you for any insight. Chantel Boswell HT (ASCP) Waco, Tx Thank you in advance, Chantel Boswell HT (ASCP) Waco, Tx _______________________________________________ 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 la.sebree <@t> hosp.wisc.edu Wed Aug 31 08:12:15 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Control Slides Message-ID: Speaking only as a tech that performs IHC and ISH, we use our own control slides whenever possible unless we have been unable to find positive control tissue, i.e. adenovirus. We not only review the positive control tissue but the patient as well just in case there is an unexpected result on the patient tissue that needs investigating. Even in cases where the control tissue is on the same slide as the patient tissue, there have been instances where one has stained appropriately and the other has not. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kevin and Chantel Bosewell Sent: Wednesday, August 31, 2005 3:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Control Slides Hello All, Our Pathologist believe control slides which are purchased are bomb proof and we should process our own. How does everyone else feel about this? Oh, and one more question, when you do any special stain do you review the patient slide or just the control? I have been asking around and I have been getting different answers. We recently had a false negative on a GMS for Fungus and this tech does not review the patient slides, in this case the control slide was positive. Thank you for any insight. Chantel Boswell HT (ASCP) Waco, Tx Thank you in advance, Chantel Boswell HT (ASCP) Waco, Tx _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Marirose.Satterfield <@t> MercyMemorial.org Wed Aug 31 08:36:10 2005 From: Marirose.Satterfield <@t> MercyMemorial.org (Satterfield, Marirose) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] sentinel biopsies Message-ID: <545CCCF1899E2042AA1C7A0DF101FC7C137358@mmh-01sv0101ew.mercymemorial.org> If anybody does cut a frozen section on the radioactive node, do you decontaminate the cryostat? What is your procedure? Thanks M. Satterfield From pex0220 <@t> yahoo.com.cn Wed Aug 31 08:37:52 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] chondrocytes Message-ID: <20050831133752.53300.qmail@web15505.mail.cnb.yahoo.com> Hello, all, I would like to do primary cultures of articular chondrocytes from newborn mice, then immunofluorescence will be performed. But I have no idea about chondrocyte cultures. Anybody can do me a favor? Thank you! Guofeng --------------------------------- DO YOU YAHOO!? ÑÅ»¢Ãâ·ÑGÓÊÏ䣭ÖйúµÚÒ»¾øÎÞÀ¬»øÓʼþɧÈų¬´óÓÊÏä From Tbarnhart <@t> primecare.org Wed Aug 31 08:47:42 2005 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] sentinel biopsies Message-ID: <1779904B5E82D511914C00D0B793339205BFD9F5@exchangent> We performed an extensive (35 specimen) study of the radioactivity levels of both the nodes and the mastectomy (lumpectomy) specimens and found the levels for the nodes were near background. At two inches from the specimen, the levels were at background. Therefore, we treat the nodes as regular frozen specimens in regard to the radioactivity. We do perform touch preps and the routine immunos on the permanents. The mastectomy and lumectomy specimens were too hot to be handled and kept in out radiology for 72 hours, remonitored and signed off as being at background levels before being sent to the histology lab. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Manager St. Alexius Medical Center Bismarck, ND -----Original Message----- From: Satterfield, Marirose [mailto:Marirose.Satterfield@MercyMemorial.org] Sent: Wednesday, August 31, 2005 8:36 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] sentinel biopsies If anybody does cut a frozen section on the radioactive node, do you decontaminate the cryostat? What is your procedure? Thanks M. Satterfield _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 GDawson <@t> dynacaremilwaukee.com Wed Aug 31 10:39:17 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] HT Certification ? Message-ID: All, I have an excellent lab assistant here that I believe would make an excellent histotech. He happens to be from Mexico where he was an MLT (a 3 year degree program there). I have his transpcripts from the college in Mexico but could anyone tell me who I could ask to see if or how many of his credits could be applied to the ROUTE 2 for HT certification from the ASCP website? This route states that with a year of experience in the histo lab and 60 semester hours of appropriate academic credit, he could sit for the HT certification test. I would really like to be able to tell this individual if some of the courses he took in Mexico will be applicable so if anyone has any suggestions, please let me know. Thanx in Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI From tpmorken <@t> labvision.com Wed Aug 31 10:58:35 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] HT Certification ? Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D09F@usca0082k08.labvision.apogent.com> Glenn, the ASCP recognizes Educational Credential Evaluators; see at www.ece.org 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 Dawson, Glen Sent: Wednesday, August 31, 2005 8:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HT Certification ? All, I have an excellent lab assistant here that I believe would make an excellent histotech. He happens to be from Mexico where he was an MLT (a 3 year degree program there). I have his transpcripts from the college in Mexico but could anyone tell me who I could ask to see if or how many of his credits could be applied to the ROUTE 2 for HT certification from the ASCP website? This route states that with a year of experience in the histo lab and 60 semester hours of appropriate academic credit, he could sit for the HT certification test. I would really like to be able to tell this individual if some of the courses he took in Mexico will be applicable so if anyone has any suggestions, please let me know. Thanx in Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DDDeltour <@t> mar.med.navy.mil Wed Aug 31 11:00:15 2005 From: DDDeltour <@t> mar.med.navy.mil (DDDeltour@mar.med.navy.mil) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] HT Certification ? Message-ID: <080566D001A3D9459FFC0A391A646C9106C4AA3B@marxchg03.mar.med.navy.mil> The best thing to do would be to contact ASCP directly. It says "regionally accredited college/university". Accredited by whom though?? Doug D -----Original Message----- From: Dawson, Glen [mailto:GDawson@dynacaremilwaukee.com] Sent: Wednesday, August 31, 2005 11:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HT Certification ? All, I have an excellent lab assistant here that I believe would make an excellent histotech. He happens to be from Mexico where he was an MLT (a 3 year degree program there). I have his transpcripts from the college in Mexico but could anyone tell me who I could ask to see if or how many of his credits could be applied to the ROUTE 2 for HT certification from the ASCP website? This route states that with a year of experience in the histo lab and 60 semester hours of appropriate academic credit, he could sit for the HT certification test. I would really like to be able to tell this individual if some of the courses he took in Mexico will be applicable so if anyone has any suggestions, please let me know. Thanx in Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From M.Prideaux <@t> sheffield.ac.uk Wed Aug 31 11:08:09 2005 From: M.Prideaux <@t> sheffield.ac.uk (Matt Prideaux) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] (no subject) Message-ID: <1125504489.4315d5e9617de@webmail.shef.ac.uk> Dear Histonetters, A colleague wishes to store cytospun cell preparations in the freezer before doing immunocytochemistry for CD31 and CD44. She would like to know whether it's better to fix the slides before freezing or after thawing them prior to staining. Also, which fixatives would you recommend? Matt -- Matt Prideaux Academic Unit of Bone Biology Division of Clinical Sciences (South) D Floor (DU20) Medical School Henry Wellcome Laboratories for Medical Research University of Sheffield Beech Hill Road Sheffield S10 2RX Tel: 0114 271 3783 Fax: 0114 271 1711 From dmarsha3 <@t> utmem.edu Wed Aug 31 11:34:35 2005 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Gulf Coast References: Message-ID: <001d01c5ae49$dfa84640$f623c084@DanaM> Good thinking Betsy. I second that. dm VAMC Memphis TN ----- Original Message ----- From: "Molinari, Betsy" To: Sent: Wednesday, August 31, 2005 5:49 AM Subject: [Histonet] Gulf Coast My thoughts and prayers to all of you that are affected by Katrina. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BRIAN.CHELACK <@t> usask.ca Wed Aug 31 10:45:45 2005 From: BRIAN.CHELACK <@t> usask.ca (Brian Chelack) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Re: Can PBS cause background Message-ID: <4315D0A9.81D67496@sask.usask.ca> Hello Carolyn; In the late 80's I did use an alternate procedure to inactivate endogenous peroxidases that included sodium borohydride. We were testing methods for antigens that were sensitive to H2O2-methanol. My memory is dim, but as I recall the procedure involved the immersion of the slides in 0.1% phenylhydrazine at 37C for 1 hour followed by immersion in 0.5 mg/ml NaBH4 for 10 minutes. In my experience bubbles under the sections were not a problem, but the treatment was tough on the sections. Phenylhydrazine is also not a very nice chemical to work with. If you are looking for a non-methonal method try using PBS + 2% H2O2 + 0.1% Na Azide. As far as PBS vs Tris based buffers is concerned, I believe the amino groups in the Tris can act to reduce nonspecific antibody binding. Over the years I have used both Tris and PBS based buffers and have seen differences in the staining intensity that is antibody specific. That is, certain antibodies do work slightly better with one buffer or the other. Ultimately the differences were not significant enough to warrant the use of one buffer over the other. We use PBS but Tris will work just fine. Brian Chelack Prairie Diagnostic Services From quinntl <@t> umkc.edu Wed Aug 31 12:09:23 2005 From: quinntl <@t> umkc.edu (Quinn, Tim L.) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] 20% glycerol 2%DMSO cryoprotection method- Effects Message-ID: <3563FA9377A5DC41BDDA1E8A6C06BEF8034D2AD6@KC-MAIL6.kc.umkc.edu> Dear Listies, How does the 20% glycerol 2%DMSO cryo-protection method affect immunolabeling of DMSO soluble cytokines? Timothy L. Quinn Senior Lab Techician Missouri University at Kansas City Medical School- Med Lab 2411 Holmes Kansas City, MO 64108 816-235-1904 quinntl@umkc.edu From vazquezr <@t> ohsu.edu Wed Aug 31 12:14:11 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Gulf Coast Message-ID: I agree Robyn OHSU >>> "Dana Marshall" 8/31/2005 9:34 AM >>> Good thinking Betsy. I second that. dm VAMC Memphis TN ----- Original Message ----- From: "Molinari, Betsy" To: Sent: Wednesday, August 31, 2005 5:49 AM Subject: [Histonet] Gulf Coast My thoughts and prayers to all of you that are affected by Katrina. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From secash <@t> gundluth.org Wed Aug 31 12:21:29 2005 From: secash <@t> gundluth.org (secash@gundluth.org) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] sentinel biopsies Message-ID: We have been told that you get more of a dose of radiation going from the lab to our cars than from sentinel lymph nodes during frozen sections. I have more important things to worry about....like gas prices!!!!! From jmahoney <@t> alegent.org Wed Aug 31 12:31:55 2005 From: jmahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:25:31 2005 Subject: [Histonet] Data Message-ID: Hello, I'm trying to find some data about what the vacancy rate for HT's is throughout the country. I'd also like to find out about how many HT's retire each year and how many registered techs come into the field every year. Any help would be greatly appreciated. Jan Omaha Janice Mahoney Histology/Cytology Coordinator Alegent Health Laboratory Phone (402)717-2889 Fax (402)717-5231 "We pledge to be creative, visionary leaders committed to holistic healthcare in the region" From KMH.02 <@t> ex.uchs.org Wed Aug 31 12:38:38 2005 From: KMH.02 <@t> ex.uchs.org (Hopkins, Karen) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] HT certification Message-ID: <640B23A5DC4B234BB065E56F2DB30596028AB06F@uchex2ucmc.uchs.org> You can get information on this as well as certificate programs from Harford Community College in Bel Air Maryland. They offer long distance programs via the internet. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, August 31, 2005 1:05 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 21, Issue 41 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. HT Certification ? (Dawson, Glen) 2. RE: HT Certification ? (Morken, Tim - Labvision) 3. RE: HT Certification ? (DDDeltour@mar.med.navy.mil) 4. (no subject) (Matt Prideaux) 5. Re: Gulf Coast (Dana Marshall) 6. Re: Can PBS cause background (Brian Chelack) ---------------------------------------------------------------------- Message: 1 Date: Wed, 31 Aug 2005 10:39:17 -0500 From: "Dawson, Glen" Subject: [Histonet] HT Certification ? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" All, I have an excellent lab assistant here that I believe would make an excellent histotech. He happens to be from Mexico where he was an MLT (a 3 year degree program there). I have his transpcripts from the college in Mexico but could anyone tell me who I could ask to see if or how many of his credits could be applied to the ROUTE 2 for HT certification from the ASCP website? This route states that with a year of experience in the histo lab and 60 semester hours of appropriate academic credit, he could sit for the HT certification test. I would really like to be able to tell this individual if some of the courses he took in Mexico will be applicable so if anyone has any suggestions, please let me know. Thanx in Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI ------------------------------ Message: 2 Date: Wed, 31 Aug 2005 11:58:35 -0400 From: "Morken, Tim - Labvision" Subject: RE: [Histonet] HT Certification ? To: "'Dawson, Glen'" , histonet@lists.utsouthwestern.edu Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D09F@usca0082k08.labvision.apogent. com> Content-Type: text/plain Glenn, the ASCP recognizes Educational Credential Evaluators; see at www.ece.org 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 Dawson, Glen Sent: Wednesday, August 31, 2005 8:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HT Certification ? All, I have an excellent lab assistant here that I believe would make an excellent histotech. He happens to be from Mexico where he was an MLT (a 3 year degree program there). I have his transpcripts from the college in Mexico but could anyone tell me who I could ask to see if or how many of his credits could be applied to the ROUTE 2 for HT certification from the ASCP website? This route states that with a year of experience in the histo lab and 60 semester hours of appropriate academic credit, he could sit for the HT certification test. I would really like to be able to tell this individual if some of the courses he took in Mexico will be applicable so if anyone has any suggestions, please let me know. Thanx in Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Wed, 31 Aug 2005 12:00:15 -0400 From: DDDeltour@mar.med.navy.mil Subject: RE: [Histonet] HT Certification ? To: GDawson@dynacaremilwaukee.com, histonet@lists.utsouthwestern.edu Message-ID: <080566D001A3D9459FFC0A391A646C9106C4AA3B@marxchg03.mar.med.navy.mil> Content-Type: text/plain The best thing to do would be to contact ASCP directly. It says "regionally accredited college/university". Accredited by whom though?? Doug D -----Original Message----- From: Dawson, Glen [mailto:GDawson@dynacaremilwaukee.com] Sent: Wednesday, August 31, 2005 11:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] HT Certification ? All, I have an excellent lab assistant here that I believe would make an excellent histotech. He happens to be from Mexico where he was an MLT (a 3 year degree program there). I have his transpcripts from the college in Mexico but could anyone tell me who I could ask to see if or how many of his credits could be applied to the ROUTE 2 for HT certification from the ASCP website? This route states that with a year of experience in the histo lab and 60 semester hours of appropriate academic credit, he could sit for the HT certification test. I would really like to be able to tell this individual if some of the courses he took in Mexico will be applicable so if anyone has any suggestions, please let me know. Thanx in Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Wed, 31 Aug 2005 17:08:09 +0100 From: Matt Prideaux Subject: [Histonet] (no subject) To: Histonet Message-ID: <1125504489.4315d5e9617de@webmail.shef.ac.uk> Content-Type: text/plain; charset=ISO-8859-1 Dear Histonetters, A colleague wishes to store cytospun cell preparations in the freezer before doing immunocytochemistry for CD31 and CD44. She would like to know whether it's better to fix the slides before freezing or after thawing them prior to staining. Also, which fixatives would you recommend? Matt -- Matt Prideaux Academic Unit of Bone Biology Division of Clinical Sciences (South) D Floor (DU20) Medical School Henry Wellcome Laboratories for Medical Research University of Sheffield Beech Hill Road Sheffield S10 2RX Tel: 0114 271 3783 Fax: 0114 271 1711 ------------------------------ Message: 5 Date: Wed, 31 Aug 2005 11:34:35 -0500 From: "Dana Marshall" Subject: Re: [Histonet] Gulf Coast To: "Molinari, Betsy" , Message-ID: <001d01c5ae49$dfa84640$f623c084@DanaM> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Good thinking Betsy. I second that. dm VAMC Memphis TN ----- Original Message ----- From: "Molinari, Betsy" To: Sent: Wednesday, August 31, 2005 5:49 AM Subject: [Histonet] Gulf Coast My thoughts and prayers to all of you that are affected by Katrina. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Wed, 31 Aug 2005 09:45:45 -0600 From: Brian Chelack Subject: [Histonet] Re: Can PBS cause background To: histonet@lists.utsouthwestern.edu Message-ID: <4315D0A9.81D67496@sask.usask.ca> Content-Type: text/plain; charset=us-ascii Hello Carolyn; In the late 80's I did use an alternate procedure to inactivate endogenous peroxidases that included sodium borohydride. We were testing methods for antigens that were sensitive to H2O2-methanol. My memory is dim, but as I recall the procedure involved the immersion of the slides in 0.1% phenylhydrazine at 37C for 1 hour followed by immersion in 0.5 mg/ml NaBH4 for 10 minutes. In my experience bubbles under the sections were not a problem, but the treatment was tough on the sections. Phenylhydrazine is also not a very nice chemical to work with. If you are looking for a non-methonal method try using PBS + 2% H2O2 + 0.1% Na Azide. As far as PBS vs Tris based buffers is concerned, I believe the amino groups in the Tris can act to reduce nonspecific antibody binding. Over the years I have used both Tris and PBS based buffers and have seen differences in the staining intensity that is antibody specific. That is, certain antibodies do work slightly better with one buffer or the other. Ultimately the differences were not significant enough to warrant the use of one buffer over the other. We use PBS but Tris will work just fine. Brian Chelack Prairie Diagnostic Services ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 21, Issue 41 **************************************** From lchung <@t> ppmh.org Wed Aug 31 12:36:13 2005 From: lchung <@t> ppmh.org (Chung, Luong) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Gulf Coast Message-ID: Prayers go out to the victims. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Robyn Vazquez Sent: Wednesday, August 31, 2005 1:14 PM To: BMolinari@heart.thi.tmc.edu; histonet@lists.utsouthwestern.edu; dmarsha3@utmem.edu Subject: Re: [Histonet] Gulf Coast I agree Robyn OHSU >>> "Dana Marshall" 8/31/2005 9:34 AM >>> Good thinking Betsy. I second that. dm VAMC Memphis TN ----- Original Message ----- From: "Molinari, Betsy" To: Sent: Wednesday, August 31, 2005 5:49 AM Subject: [Histonet] Gulf Coast My thoughts and prayers to all of you that are affected by Katrina. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sharon.osborn <@t> dnax.org Wed Aug 31 15:54:28 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Job listings Message-ID: <29B25753F6B1D51196110002A589D44402398195@PALMSG30.us.schp.com> Histonetters, In the next few days we will have postings on our website for several positions in histology with IHC, insitu, LCM, TMA, computer experience. Please check www.dnaxresearch.com for those descriptions. We are hiring for 2-4 positions with varying responsibilities. After the job descriptions are posted, you can apply online or send your resume to Martha Hauser, Director of Personnel, SP BioPharma, 901 California, Palo Alto, CA 94304. ScheringPlough BioPharma (DNAX) is a biotechnology company located within the San Franciso Bay Area Biotechnology sector and adjacent to Stanford University. SP BioPharma carries out translational research leading to drug development in areas of Immunology and Oncology in a highly collaborative environment. I am off to Plainview Texas to settle personal effects of my uncle's estate, returning Sept 12th. I can respond more after return. Sharon Osborn, Histology DNAX, SP BioPharma 901 California Palo Alto, CA 650.496.6539 650.278.1944 cell ********************************************************************* 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 sv18 <@t> columbia.edu Wed Aug 31 16:51:57 2005 From: sv18 <@t> columbia.edu (Sunilda Valladares-Silva) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] 4 histology technician positions plus technical supervisory position available. Message-ID: <4316267D.8050109@columbia.edu> We currently have opening for 4 full time Histology Technician and a Technical Supervisor in a high volume clinical laboratory at Columbia University Medical Center, New York City. The positions requires individuals to work on the night shift: handling all surgical biopsies. Certification and Bachelor of Science required. Salaries compensated with education and experience. If interested and would like more details, please contact Sunny Silva at 212-305-8263 or Doreen Hebert, 212-305-4886. Forward cv to 212-305-2301. From chapcl <@t> yahoo.com Wed Aug 31 17:09:31 2005 From: chapcl <@t> yahoo.com (William Chappell) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Gulf Coast and Volunteerism Message-ID: <20050831220931.77949.qmail@web60023.mail.yahoo.com> Does any one know if any of the field hospitals have pathology? Is there a need for a HTL in the hurricane affected areas. Maybe a deanor or at least someone who is not squimish around dead bodies. I am interested in volunteering for as long as needed. William, HTL From jnocito <@t> satx.rr.com Wed Aug 31 17:10:58 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Control Slides References: <000601c5ae07$94eef6e0$c9fb5a42@Rascal> Message-ID: <00cd01c5ae78$de1e8da0$52bf0b43@yourxhtr8hvc4p> Chantel, we try to use tissues and cases that we find positive that were processed in our lab. However, we have had to purchase control slides from companies because we just don't have them and the pathologsits' do not want to spend the money sending them to another lab. We do look at the "control slide" that is sent for each batch to make sure we get what we pay for. As for looking at the patient slides, we always are looking to see if we can use that block or tissue as a control for ourselves, whether it is for special stains or immunos. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Kevin and Chantel Bosewell" To: Sent: Wednesday, August 31, 2005 3:39 AM Subject: [Histonet] Control Slides Hello All, Our Pathologist believe control slides which are purchased are bomb proof and we should process our own. How does everyone else feel about this? Oh, and one more question, when you do any special stain do you review the patient slide or just the control? I have been asking around and I have been getting different answers. We recently had a false negative on a GMS for Fungus and this tech does not review the patient slides, in this case the control slide was positive. Thank you for any insight. Chantel Boswell HT (ASCP) Waco, Tx Thank you in advance, Chantel Boswell HT (ASCP) Waco, Tx _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Anti-Virus. Version: 7.0.344 / Virus Database: 267.10.17/85 - Release Date: 8/30/2005 From jnocito <@t> satx.rr.com Wed Aug 31 17:15:23 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Gulf Coast References: Message-ID: <00db01c5ae79$7c018f40$52bf0b43@yourxhtr8hvc4p> being through a couple of hurricanes when I was in Miami compares to nothing like what happened this week. My thoughts and prayers are with everyone in the Gulf states. I did my histology training in Biloxi MS and have been to New Orleans many times. Such a catastrophe. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Robyn Vazquez" To: ; ; Sent: Wednesday, August 31, 2005 12:14 PM Subject: Re: [Histonet] Gulf Coast I agree Robyn OHSU >>> "Dana Marshall" 8/31/2005 9:34 AM >>> Good thinking Betsy. I second that. dm VAMC Memphis TN ----- Original Message ----- From: "Molinari, Betsy" To: Sent: Wednesday, August 31, 2005 5:49 AM Subject: [Histonet] Gulf Coast My thoughts and prayers to all of you that are affected by Katrina. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Anti-Virus. Version: 7.0.344 / Virus Database: 267.10.17/85 - Release Date: 8/30/2005 From akarakou <@t> internode.on.net Wed Aug 31 18:45:24 2005 From: akarakou <@t> internode.on.net (Internode) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Re: Histonet Digest, Vol 21, Issue 41 References: <1125507619_32904@mail.internode.on.net> Message-ID: <000401c5ae86$128f2970$6be17acb@clientor7dajn2> Got the interview tomorrow ----- Original Message ----- From: To: Sent: Thursday, September 01, 2005 2:30 AM Subject: Histonet Digest, Vol 21, Issue 41 > 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. HT Certification ? (Dawson, Glen) > 2. RE: HT Certification ? (Morken, Tim - Labvision) > 3. RE: HT Certification ? (DDDeltour@mar.med.navy.mil) > 4. (no subject) (Matt Prideaux) > 5. Re: Gulf Coast (Dana Marshall) > 6. Re: Can PBS cause background (Brian Chelack) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Wed, 31 Aug 2005 10:39:17 -0500 > From: "Dawson, Glen" > Subject: [Histonet] HT Certification ? > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > All, > > I have an excellent lab assistant here that I believe would make an > excellent histotech. He happens to be from Mexico where he was an MLT (a > 3 > year degree program there). I have his transpcripts from the college in > Mexico but could anyone tell me who I could ask to see if or how many of > his > credits could be applied to the ROUTE 2 for HT certification from the ASCP > website? This route states that with a year of experience in the histo > lab > and 60 semester hours of appropriate academic credit, he could sit for the > HT certification test. > > I would really like to be able to tell this individual if some of the > courses he took in Mexico will be applicable so if anyone has any > suggestions, please let me know. > > Thanx in Advance, > > Glen Dawson BS, HT & QIHC (ASCP) > IHC Manager > Milwaukee, WI > > > > > > > ------------------------------ > > Message: 2 > Date: Wed, 31 Aug 2005 11:58:35 -0400 > From: "Morken, Tim - Labvision" > Subject: RE: [Histonet] HT Certification ? > To: "'Dawson, Glen'" , > histonet@lists.utsouthwestern.edu > Message-ID: > <0556BE8AC5551E4E8AF6BB9E42509BA20328D09F@usca0082k08.labvision.apogent.com> > > Content-Type: text/plain > > Glenn, the ASCP recognizes Educational Credential Evaluators; see at > www.ece.org > > 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 Dawson, > Glen > Sent: Wednesday, August 31, 2005 8:39 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] HT Certification ? > > > All, > > I have an excellent lab assistant here that I believe would make an > excellent histotech. He happens to be from Mexico where he was an MLT (a > 3 > year degree program there). I have his transpcripts from the college in > Mexico but could anyone tell me who I could ask to see if or how many of > his > credits could be applied to the ROUTE 2 for HT certification from the ASCP > website? This route states that with a year of experience in the histo > lab > and 60 semester hours of appropriate academic credit, he could sit for the > HT certification test. > > I would really like to be able to tell this individual if some of the > courses he took in Mexico will be applicable so if anyone has any > suggestions, please let me know. > > Thanx in Advance, > > Glen Dawson BS, HT & QIHC (ASCP) > IHC Manager > Milwaukee, WI > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ------------------------------ > > Message: 3 > Date: Wed, 31 Aug 2005 12:00:15 -0400 > From: DDDeltour@mar.med.navy.mil > Subject: RE: [Histonet] HT Certification ? > To: GDawson@dynacaremilwaukee.com, histonet@lists.utsouthwestern.edu > Message-ID: > <080566D001A3D9459FFC0A391A646C9106C4AA3B@marxchg03.mar.med.navy.mil> > Content-Type: text/plain > > The best thing to do would be to contact ASCP directly. It says > "regionally > accredited college/university". Accredited by whom though?? > Doug D > > -----Original Message----- > From: Dawson, Glen [mailto:GDawson@dynacaremilwaukee.com] > Sent: Wednesday, August 31, 2005 11:39 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] HT Certification ? > > All, > > I have an excellent lab assistant here that I believe would make an > excellent histotech. He happens to be from Mexico where he was an MLT (a > 3 > year degree program there). I have his transpcripts from the college in > Mexico but could anyone tell me who I could ask to see if or how many of > his > credits could be applied to the ROUTE 2 for HT certification from the ASCP > website? This route states that with a year of experience in the histo > lab > and 60 semester hours of appropriate academic credit, he could sit for the > HT certification test. > > I would really like to be able to tell this individual if some of the > courses he took in Mexico will be applicable so if anyone has any > suggestions, please let me know. > > Thanx in Advance, > > Glen Dawson BS, HT & QIHC (ASCP) > IHC Manager > Milwaukee, WI > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > ------------------------------ > > Message: 4 > Date: Wed, 31 Aug 2005 17:08:09 +0100 > From: Matt Prideaux > Subject: [Histonet] (no subject) > To: Histonet > Message-ID: <1125504489.4315d5e9617de@webmail.shef.ac.uk> > Content-Type: text/plain; charset=ISO-8859-1 > > Dear Histonetters, > > A colleague wishes to store cytospun cell preparations in the freezer > before > doing immunocytochemistry for CD31 and CD44. She would like to know > whether > it's better to fix the slides before freezing or after thawing them prior > to > staining. Also, which fixatives would you recommend? > > Matt > -- > Matt Prideaux > Academic Unit of Bone Biology > Division of Clinical Sciences (South) > D Floor (DU20) Medical School > Henry Wellcome Laboratories for Medical Research > University of Sheffield > Beech Hill Road > Sheffield > S10 2RX > > Tel: 0114 271 3783 > Fax: 0114 271 1711 > > > > > > > > ------------------------------ > > Message: 5 > Date: Wed, 31 Aug 2005 11:34:35 -0500 > From: "Dana Marshall" > Subject: Re: [Histonet] Gulf Coast > To: "Molinari, Betsy" , > > Message-ID: <001d01c5ae49$dfa84640$f623c084@DanaM> > Content-Type: text/plain; format=flowed; charset="iso-8859-1"; > reply-type=original > > Good thinking Betsy. > I second that. > dm > VAMC Memphis TN > > ----- Original Message ----- > From: "Molinari, Betsy" > To: > Sent: Wednesday, August 31, 2005 5:49 AM > Subject: [Histonet] Gulf Coast > > > My thoughts and prayers to all of you that are affected by Katrina. > > > > Betsy Molinari HT (ASCP) > > Texas Heart Institute > > Cardiovascular Pathology > > 6770 Bertner Ave. > > Houston,TX 77030 > > 832-355-6524 > > 832-355-6812 (fax) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > ------------------------------ > > Message: 6 > Date: Wed, 31 Aug 2005 09:45:45 -0600 > From: Brian Chelack > Subject: [Histonet] Re: Can PBS cause background > To: histonet@lists.utsouthwestern.edu > Message-ID: <4315D0A9.81D67496@sask.usask.ca> > Content-Type: text/plain; charset=us-ascii > > Hello Carolyn; > > In the late 80's I did use an alternate procedure to > inactivate endogenous peroxidases that included sodium > borohydride. We were testing methods for antigens that were > sensitive to H2O2-methanol. My memory is dim, but as I > recall the procedure involved the immersion of the slides in > 0.1% phenylhydrazine at 37C for 1 hour followed by immersion > in 0.5 mg/ml NaBH4 for 10 minutes. In my experience bubbles > under the sections were not a problem, but the treatment was > tough on the sections. Phenylhydrazine is also not a very > nice chemical to work with. If you are looking for a > non-methonal method try using PBS + 2% H2O2 + 0.1% Na Azide. > > As far as PBS vs Tris based buffers is concerned, I believe > the amino groups in the Tris can act to reduce nonspecific > antibody binding. Over the years I have used both Tris and > PBS based buffers and have seen differences in the staining > intensity that is antibody specific. That is, certain > antibodies do work slightly better with one buffer or the > other. Ultimately the differences were not significant > enough to warrant the use of one buffer over the other. We > use PBS but Tris will work just fine. > > Brian Chelack > Prairie Diagnostic Services > > > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 21, Issue 41 > **************************************** From ernestinemiddleton <@t> yahoo.ca Wed Aug 31 19:41:25 2005 From: ernestinemiddleton <@t> yahoo.ca (Ernestine Middleton) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Position opening Message-ID: <20050901004125.11994.qmail@web51504.mail.yahoo.com> Montefiore Medical Center, Bronx, NY has an opening for Clinical Histology Technician. Primary duties are cutting, embedding, maintenance of tissue processor. An Associate degree is the minimun education background. Call Ernestine Middleton at 718-920-4157 or fax resume to 718-547-1920 __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From RJLevier <@t> LancasterGeneral.org Wed Aug 31 20:53:19 2005 From: RJLevier <@t> LancasterGeneral.org (LeVier, Rebecca J) Date: Fri Sep 16 15:25:32 2005 Subject: [Histonet] Histology Tech Opening Message-ID: <0FDBF29200C637468CCC1F9E7E85A3D214E8A1@MAIL-LR.lha.org> Hi HistoNet Folks, There is a Histology Tech opening at Lancaster General Hospital in Lancaster, Pennsylvania. It is a day shift position. If anyone is interested please check out their website. Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of 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.