RE: [Histonet] Masson Trichome Staining: Can I use Fast Green?
We use 5 minutes on FG. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of cscam...@uci.edu Sent: Thursday, April 08, 2010 6:42 PM To: HistoNet Subject: [Histonet] Masson Trichome Staining: Can I use Fast Green? Hi Histonet, I am currently using the protocol from Stainsfile for staining heart tissue: http://stainsfile.info/StainsFile/stain/conektv/masson.htm My lab had Fast Green readily available so I substituted it for Light Green in Solution C. The slides have not been coming out well. I failed to account for the difference in timing between FG and LG - running the slide in FG for the full 10 minutes. Has anyone used FG in Masson's Trichrome? If so, how long did it take to get the desired colors in the solution? Or, is Fast Green a poor substitute for Light Green? Thanks for your advice! -Colin ___ Histonet mailing list Histonet@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] Immuno staining on plastic sections
We are trying to do immuno staining on plastic sections and are not having much luck. Does anyone have any experience with this? We are embedding in technovit 8100. It is a device with encapsulated VEGFR cells. I have been told that we might have to do an etching step. Any help would be greatly appreciated. Bruce ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cd31 in pigs.
Does anyone have a procedure that WORKS for cd31 in pigs? I can't seem to get it to work no matter what I do. Any help will be greatly appreciated. Thanks.- joel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Does a Cytotechnologist Meet CAP Grossing Requirements?
Good Morning to All and Happy Friday! In your opinion, does a graduate from a Cytotechnologist Program (ASCP) with additional documented training meet the new requirement for grossing as high complexity testing personnel under CLIA??? Our PA that works for our Pathologist group is a cytotech who did additional training in grossing. Thanks, Wanda WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC 29406 843-847-4586 843-847-4296 fax 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] CPT coding
They are two distinct specimens, if you diagnose the two separately you may bill for two separate specimens even though they were in one container. Jan M Omaha, NE Sponsored by Catholic Health Initiatives and Immanuel Health Systems, Alegent Health is faithful to the healing ministry of Jesus Christ, providing high quality care for the body, mind and spirit of every person. The information contained in this communication, including attachments, is confidential and private and intended only for the use of the addressees. Unauthorized use, disclosure, distribution or copying is strictly prohibited and may be unlawful. If you received this communication in error, please inform us of the erroneous delivery by return e-mail message from your computer. Additionally, although all attachments have been scanned at the source for viruses, the recipient should check any attachments for the presence of viruses before opening. Alegent Health accepts no liability for any damage caused by any virus transmitted by this e-mail. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] CPT coding
We do a surgical examination on the placenta and the fetus when they are in the same container. We charge 88307 for the placenta and 88309 for the fetus. Thanks, Wanda WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC 29406 843-847-4586 843-847-4296 fax 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. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of histot...@imagesbyhopper.com Sent: Thursday, April 08, 2010 9:32 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CPT coding Hi Histonetters! We have run into a question about CPT charges and I was hoping to get a more definitive answer than what we currently have. I know if you have two tonsils in one container, one with a stitch and one without, you can charge for both tonsils. How about if you have one container with a placenta and a stillborn fetus? One suggestion is to charge an 88307 and the other is to charge 88300 AND an 88307 (to cover the gross only on the fetus and the tissue that was submitted for the placenta). What is the standard, shall I say legal?, way of charging for these specimens? Must you choose one of the charges or are we allowed to use both charges? Thanks for any light you can shed on this! Michelle ___ Histonet mailing list Histonet@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] RE: Does a Cytotechnologist Meet CAP Grossing Requirements?
Wanda, If they do not have a BS as well as their CT(ASCP) they need to have documentation of the required number of hours of biology and chemistry in addition to the training. If so, you should be ok. Jan Mahoney Omaha, NE -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of wanda.sm...@hcahealthcare.com Sent: Friday, April 09, 2010 8:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Does a Cytotechnologist Meet CAP Grossing Requirements? Good Morning to All and Happy Friday! In your opinion, does a graduate from a Cytotechnologist Program (ASCP) with additional documented training meet the new requirement for grossing as high complexity testing personnel under CLIA??? Our PA that works for our Pathologist group is a cytotech who did additional training in grossing. Thanks, Wanda WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC 29406 843-847-4586 843-847-4296 fax 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Sponsored by Catholic Health Initiatives and Immanuel Health Systems, Alegent Health is faithful to the healing ministry of Jesus Christ, providing high quality care for the body, mind and spirit of every person. The information contained in this communication, including attachments, is confidential and private and intended only for the use of the addressees. Unauthorized use, disclosure, distribution or copying is strictly prohibited and may be unlawful. If you received this communication in error, please inform us of the erroneous delivery by return e-mail message from your computer. Additionally, although all attachments have been scanned at the source for viruses, the recipient should check any attachments for the presence of viruses before opening. Alegent Health accepts no liability for any damage caused by any virus transmitted by this e-mail. Thank you for your cooperation. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] charge for pin 4 cocktail
What are people charging Pin 4 p63,CK5-14and P504S..88342 x 3 or 4 Cheryl Miller HT ASCP cm Histology Supervisor Physicians Laboratory Services Omaha, NE. 402 731 4148 PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] charge for pin 4 cocktail
x3 only because you can't tell the difference between these basal cell markers CK5 and CK14 (they stain the same cells and both in the cytoplasm). Mark Tarango On Fri, Apr 9, 2010 at 8:31 AM, Cheri Miller cmil...@physlab.com wrote: What are people charging Pin 4 p63,CK5-14and P504S..88342 x 3 or 4 Cheryl Miller HT ASCP cm Histology Supervisor Physicians Laboratory Services Omaha, NE. 402 731 4148 PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. ___ Histonet mailing list Histonet@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] RE: charge for pin 4 cocktail
X 3 because two of the markers cannot be distinguished. j -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cheri Miller Sent: Friday, April 09, 2010 11:32 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] charge for pin 4 cocktail What are people charging Pin 4 p63,CK5-14and P504S..88342 x 3 or 4 Cheryl Miller HT ASCP cm Histology Supervisor Physicians Laboratory Services Omaha, NE. 402 731 4148 PRIVILEGED / CONFIDENTIAL INFORMATION may be contained in this message. If you are not the addressee intended / indicated or agent responsible for delivering it to the addressee, you are hereby notified that you are in possession of confidential and privileged information. Any dissemination, distribution, or copying of this e-mail is strictly prohibited. If you have received this message in error, please notify the sender immediately and delete this email from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] CPT coding question
Michelle, the example that you raised of having two tonsils, one with a stitch, isn't quite the same as your problem. The two tonsils would otherwise not be identifiable as left or right. That is not a problem with a fetus and a placenta. Each is separately identifiable and each should be coded separately. You will simplify any coding review if each is a separate line on your report. Bill Tench Associate Dir. Laboratory Services Chief, Cytology Services Palomar Medical Center 555 E. Valley Parkway Escondido, California 92025 bill.te...@pph.org Voice: 760- 739-3037 Fax: 760-739-2604 mail2.pph.org made the following annotations - Confidential E-Mail: This e-mail is intended only for the person or entity to which it is addressed, and may contain information that is privileged, confidential, or otherwise protected from disclosure. Dissemination, distribution, or copying of this e-mail or the information herein by anyone other than the intended recipient is prohibited. If you have received this e-mail in error, please notify the sender by reply e-mail, and destroy the original message and all copies. - ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cd31 in pigs
Hi Joel, we have tried a few antibodies for CD31 in FFPE tissues in pigs and have had no luck. We currently use an antibody to vWF to stain pig endothelium. If someone has a working technique for CD31 in swine, I'd be happy to hear about it. Jean-Martin __ Jean-Martin Lapointe, DMV, MS, dACVP AccelLAB Inc Québec, Canada J7H 1N8 jm.lapoi...@accellab.com Date: Fri, 9 Apr 2010 09:40:00 -0400 From: Joel Israel jo...@mcclainlab.com Does anyone have a procedure that WORKS for cd31 in pigs? I can't seem to get it to work no matter what I do. Any help will be greatly appreciated. Thanks.- joel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Masson Trichome Staining: Can I use Fast Green?
The slides have not been coming out well. is not very informative! Your problems may have nothing to do with which dye you are using as the collagen stain. There are plenty of other things that can make a multi-step trichrome method give unexpected results. Fast green FCF (C.I. 42053) is usually considered superior to light green SF (C.I. 42095) on account of being less prone to fading. Be sure you have the correct dye, and that it is from a batch certified by the Biological Stain Commission (B.S.C. - see the label on the bottle of dye powder, which will also tell you the dye content). If you buy a ready-made solution, check with the supplier. The minimum acceptable dye content for certification of fast green FCF (85%) is higher than the minimum for light green SF (65%). A 2% solution of the former might therefore have greater tinctorial power than a 2% solution of the latter. The B.S.C. tests both dyes as substitutes for aniline blue in Gomori's one-step trichrome method; the concentration of blue or green dye in the mixture is 0.3%, irrespective of the dye content of the powder. Gomori's is a more stringent test for dyes than Masson's trichrome method because there is no visual/manual control of the staining. A dye that works with Gomori's method should work well with Masson's. The Masson variant that you have been trying (from Bryan Llewellyn's excellent web site, http://stainsfile.info/StainsFile/stain/conektv/masson.htm) is well documented. Follow up some references from Bryan's citation of the Bancroft Stevens book, and learn the reasons for all the 11 steps of the modified Masson technique. The current edition (6th, 2008) of Theory and Practice is edited by Bancroft Gamble: ISBN 9780443102790. My textbook (ISBN 97819048422) covers trichrome methods in Chapter 8, with plenty of references to scientific/scholarly literature, much of which is now available on the internet, especially if you can go through a university or public library's web interface. There is a well ilustrated chapter on troubleshooting trichromes, by Vinnie Della Speranza, in R.W.Brown's (2009) Common Problems book, ISBN 9780930304959 (pp. 95-101). A rinse in tap water at any stage after after washing out the iron-haematoxylin nuclear stain can weaken either the red or the blue/green component of any trichrome method. So can graded alcohols for the final dehydration of well stained slides. John Kiernan Anatomy, UWO London, Canada = = = - Original Message - From: cscam...@uci.edu Date: Thursday, April 8, 2010 18:42 Subject: [Histonet] Masson Trichome Staining: Can I use Fast Green? To: HistoNet histonet@lists.utsouthwestern.edu Hi Histonet, I am currently using the protocol from Stainsfile for staining heart tissue: http://stainsfile.info/StainsFile/stain/conektv/masson.htm My lab had Fast Green readily available so I substituted it for Light Green in Solution C. The slides have not been coming out well. I failed to account for the difference in timing between FG and LG - running the slide in FG for the full 10 minutes. Has anyone used FG in Masson's Trichrome?If so, how long did it take to get the desired colors in the solution? Or, is Fast Green a poor substitute for Light Green? Thanks for your advice! -Colin ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Immuno staining on plastic sections
A few years ago I took a workshop at the NSH presented by Neil Hand. He basically provided an account of his experience in working with something like over 100 antibodies used on resin (MMA) embedded bone and possibly other tissues. Maybe you can look his information up on the NSH website and contact him directly? With regards to an etching step, there are several posts that describe everything from HCl to formic acid to sodium ethoxide. In fact, I just found this posting from Gayle Callis back in 2004. Hope this helps! Jack [Histonet] Repy on immunostaining with Technovit 8100 / glycol methacrylate Gayle Callis gcallis @t montana.edu Thu Aug 12 10:13:55 CDT 2004 GMA is not a very ideal embedding media for immunohistochemistry, you would be far better off with paraffin using this antibody. GMA cannot be removed once polymerized nor sodium ethoxide (generally reserved for electron microscopy resins) etched with much success, you would be better off embedding in methylmethacrylate and remove the plastic entirely. This has been done with great success by Neil Hand (he has publications) using warm xylene, but he used stringent pressure cooker retrieval and worked with human tissue. The problem with GMA is it prevents the immunoglobulins from reaching antigenic sites, as the GMA is hydrophobic plus it can't be removed once polymerized. It will soften in the presence of water. Also, as GMA polymerizes it becomes very hot due to exothermic reaction unless you control this temperature by letting your blocks polymerize on ice in a refrigerator?? GMA with IHC problems have been discussed at length on Histonet many times, go to Histonet archives and search at www.histosearch.org. Personally, I don't think the product suggestion is correct, as it only suggests but does not say it WILL work. That is probably the reason you don't find it in the literature! Many people have experienced failure of IHC on GMA embedded tissues. I think some have had success with immunofluroescence using immunoglobulins and not a lot of antibodies either. It was a tedious stringent protocol described in a symposium talk. I think you will find in Histonet commentary, that most people attempting IHC/GMA suggest going to another embedding media. Also, CD68 ED1 is a Mouse antiRat, rather than a rat antiMouse (clone FA-11). ED1 and other rat macrophage markers, ED2, ED3, have been discussed on Histonet as FFPE tissues including retrieval for IHC. The staining was very straightforward as was retrieval described and solvents used plus heat of paraffin processing did not damage antigen. ED1, when used in on rat tissue, works well on paraffin sections, although we prefer frozen sections to avoid all aldehyde fixation and no retrieval. When we do frozen sections, the ED1 is diluted out 1:3000 or so, it will not be so dilute for paraffin sections and we detected with secondary to mouse IgG1 isotype (Southern Biotechnology) SA-HRP, and AEC chromogen. Staining pattern is spectacular in a rat spleen, normal positive control. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 To: histonet@lists.utsouthwestern.edu Date: Fri, 9 Apr 2010 09:11:59 -0400 From: brusk...@aol.com Subject: [Histonet] Immuno staining on plastic sections We are trying to do immuno staining on plastic sections and are not having much luck. Does anyone have any experience with this? We are embedding in technovit 8100. It is a device with encapsulated VEGFR cells. I have been told that we might have to do an etching step. Any help would be greatly appreciated. Bruce ___ Histonet mailing list Histonet@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] IHC on FNA
Happy Friday Histonetters What is everyone using for controls for FNA IHC stains. We routinely stain human FFPE sections and only have controls for tissue. Recently we had a few cases when the only sample was an FNA to perform IHC on. Any suggestions would be welcome . Regards Cindy Pyse, CLT, HT (ASCP) Histology Supervisor X-Cell Laboratories e-mail cp...@x-celllab.com ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Cd31 in pigs
Von Willebrand Factor (Factor VIII) is also what I use to stain pig endothelium. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive...@umn.edu (Confidentiality Notice: This message, together with any attachments, is intended only for the use of the individual or entity to which it is addressed and may contain confidential or privileged information. If you think you have received this message in error, please advise the sender and then delete this message and any attachments immediately.) - Original Message - From: Jean-Martin Lapointe jm.lapoi...@accellab.com To: histonet@lists.utsouthwestern.edu Sent: Friday, April 09, 2010 10:59 AM Subject: [Histonet] Cd31 in pigs Hi Joel, we have tried a few antibodies for CD31 in FFPE tissues in pigs and have had no luck. We currently use an antibody to vWF to stain pig endothelium. If someone has a working technique for CD31 in swine, I'd be happy to hear about it. Jean-Martin __ Jean-Martin Lapointe, DMV, MS, dACVP AccelLAB Inc Québec, Canada J7H 1N8 jm.lapoi...@accellab.com Date: Fri, 9 Apr 2010 09:40:00 -0400 From: Joel Israel jo...@mcclainlab.com Does anyone have a procedure that WORKS for cd31 in pigs? I can't seem to get it to work no matter what I do. Any help will be greatly appreciated. Thanks.- joel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Masson Trichome Staining: Can I use Fast Green?
** Proprietary ** ** Reply Requested When Convenient ** Colin, If you havin difficulty with MT try the Gomori One Step 1. Sections to water. 2. Pretreat in 3% Potassium dichromate for 1hr at 60ºC 3. Stain nuclei with celestin blue/Mayer's haematoxylin. 5 min 4. Wash in tap water. 5. Differentiate and blue. 6. Stain with chromotrope green mixture --- 10 mins. 7. Rinse in distilled water 8. Blot and dehydrate from absolute alcohol ,clear and mount. Results Muscle and fibrin - Red RBC's - Red Collagen- Green MASSON TRICHROME (ONE-STEP) Ingredients: Chromotrope 2R 0.6 g. Fast Green FCF 0.3 g. Phosphotungstic acid0.6 g. Glacial acetic acid 1 ml. Distilled water 100 ml. METHOD:- Dissolve the chromotrope 2R, the fast green and the phosphotungstic acid in the water and add the glacial acetic acid. Hope this help Malika Benatti Specialist BMS Camelia Botnar Laboratories Histopathology Department Great Ormond Street Hospital London WC1N 3JH Tel: +44 20 7405 9200 ext 5475 Fax: +44 20 7829 7875 ben...@gosh.nhs.uk John Kiernan jkier...@uwo.ca 09/04/2010 17:08 The slides have not been coming out well. is not very informative! Your problems may have nothing to do with which dye you are using as the collagen stain. There are plenty of other things that can make a multi-step trichrome method give unexpected results. Fast green FCF (C.I. 42053) is usually considered superior to light green SF (C.I. 42095) on account of being less prone to fading. Be sure you have the correct dye, and that it is from a batch certified by the Biological Stain Commission (B.S.C. - see the label on the bottle of dye powder, which will also tell you the dye content). If you buy a ready-made solution, check with the supplier. The minimum acceptable dye content for certification of fast green FCF (85%) is higher than the minimum for light green SF (65%). A 2% solution of the former might therefore have greater tinctorial power than a 2% solution of the latter. The B.S.C. tests both dyes as substitutes for aniline blue in Gomori's one-step trichrome method; the concentration of blue or green dye in the mixture is 0.3%, irrespective of the dye content of the powder. Gomori's is a more stringent test for dyes than Masson's trichrome method because there is no visual/manual control of the staining. A dye that works with Gomori's method should work well with Masson's. The Masson variant that you have been trying (from Bryan Llewellyn's excellent web site, http://stainsfile.info/StainsFile/stain/conektv/masson.htm) is well documented. Follow up some references from Bryan's citation of the Bancroft Stevens book, and learn the reasons for all the 11 steps of the modified Masson technique. The current edition (6th, 2008) of Theory and Practice is edited by Bancroft Gamble: ISBN 9780443102790. My textbook (ISBN 97819048422) covers trichrome methods in Chapter 8, with plenty of references to scientific/scholarly literature, much of which is now available on the internet, especially if you can go through a university or public library's web interface. There is a well ilustrated chapter on troubleshooting trichromes, by Vinnie Della Speranza, in R.W.Brown's (2009) Common Problems book, ISBN 9780930304959 (pp. 95-101). A rinse in tap water at any stage after after washing out the iron-haematoxylin nuclear stain can weaken either the red or the blue/green component of any trichrome method. So can graded alcohols for the final dehydration of well stained slides. John Kiernan Anatomy, UWO London, Canada = = = - Original Message - From: cscam...@uci.edu Date: Thursday, April 8, 2010 18:42 Subject: [Histonet] Masson Trichome Staining: Can I use Fast Green? To: HistoNet histonet@lists.utsouthwestern.edu Hi Histonet, I am currently using the protocol from Stainsfile for staining heart tissue: http://stainsfile.info/StainsFile/stain/conektv/masson.htm My lab had Fast Green readily available so I substituted it for Light Green in Solution C. The slides have not been coming out well. I failed to account for the difference in timing between FG and LG - running the slide in FG for the full 10 minutes. Has anyone used FG in Masson's Trichrome?If so, how long did it take to get the desired colors in the solution? Or, is Fast Green a poor substitute for Light Green? Thanks for your advice! -Colin ___ Histonet mailing list Histonet@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 message may contain confidential information. If you are not the
[Histonet] enzymes significance?
dear histonetters as i'm learning HC IHC i need to know a reference or internet site that could help me to know the significance of different enzymes in tissues?? any help in these!? a second quistion please i'm trying to locat Alk. Phosphatase in chicken intestine and i found in old reference the steps and reagent used for demonstration of it. i need to know is there a commercially avaliable kits ??? please share me your protocols any help will be appreciated thanx mohamed ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC on FNA
This topic touches a nerve with me. We recently went back to receiving requests on FNAs after years of not doing any. When we were routinely staining them with IHC, we would use a frozen tissue section for a control as the same protocols generally worked for both, i.e. no HIER, protease, etc as may be needed for FFPE. When we stopped doing FNA IHC routinely, we eliminated our frozen section control inventory. Now that we're doing them again, it was decided to use FFPE controls and their accompanying IHC protocols. I do not agree with this practice as I feel there are too many differences between FNA preparations and FFPE sections. I was voted down so now we are using HIER, protease, etc. whatever the FFPE protocol calls for on these FNA preparations. Surprisingly, at least to me, there are still cells left on the FNA preps after even the harshest retrieval protocols. We also don't run negative controls of the FNAs. The argument being that there are totally different cells from one slide to the next so a comparison between a negative control slide and one that received antibody is useless. So that's what we're doing even though I don't agree with it. At the time this all came about, I also queried Histonet as to common practices so the archives should have some info. also. Linda A. Sebree University of Wisconsin Hospital Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cynthia Pyse Sent: Friday, April 09, 2010 11:16 AM To: 'Histonet' Subject: [Histonet] IHC on FNA Happy Friday Histonetters What is everyone using for controls for FNA IHC stains. We routinely stain human FFPE sections and only have controls for tissue. Recently we had a few cases when the only sample was an FNA to perform IHC on. Any suggestions would be welcome . Regards Cindy Pyse, CLT, HT (ASCP) Histology Supervisor X-Cell Laboratories e-mail cp...@x-celllab.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
RE: [Histonet] IHC on FNA
Great points, Linda! -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sebree Linda A Sent: Friday, April 09, 2010 11:40 AM To: Cynthia Pyse; Histonet Subject: RE: [Histonet] IHC on FNA This topic touches a nerve with me. We recently went back to receiving requests on FNAs after years of not doing any. When we were routinely staining them with IHC, we would use a frozen tissue section for a control as the same protocols generally worked for both, i.e. no HIER, protease, etc as may be needed for FFPE. When we stopped doing FNA IHC routinely, we eliminated our frozen section control inventory. Now that we're doing them again, it was decided to use FFPE controls and their accompanying IHC protocols. I do not agree with this practice as I feel there are too many differences between FNA preparations and FFPE sections. I was voted down so now we are using HIER, protease, etc. whatever the FFPE protocol calls for on these FNA preparations. Surprisingly, at least to me, there are still cells left on the FNA preps after even the harshest retrieval protocols. We also don't run negative controls of the FNAs. The argument being that there are totally different cells from one slide to the next so a comparison between a negative control slide and one that received antibody is useless. So that's what we're doing even though I don't agree with it. At the time this all came about, I also queried Histonet as to common practices so the archives should have some info. also. Linda A. Sebree University of Wisconsin Hospital Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cynthia Pyse Sent: Friday, April 09, 2010 11:16 AM To: 'Histonet' Subject: [Histonet] IHC on FNA Happy Friday Histonetters What is everyone using for controls for FNA IHC stains. We routinely stain human FFPE sections and only have controls for tissue. Recently we had a few cases when the only sample was an FNA to perform IHC on. Any suggestions would be welcome . Regards Cindy Pyse, CLT, HT (ASCP) Histology Supervisor X-Cell Laboratories e-mail cp...@x-celllab.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
[Histonet] PT or as need work in WI/ILL
Good afternoon all, Just my periodic post looking for work in the So.WI/No.Ill area. Steve ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Job in Little Rock
Hi All, UAMS (University of Arkansas for Medical Sciences) is looking for a full time histologist. Canidate must be a regesterd HT or HTL or eligible for ASCP registry for our Histology Laboratory. It is on the UAMS website under Special Procedures Tech in the Technical area. If you are interested please fill out an application or contact me directly. Recuiters please note we are not allowed to use outside services. Pamela Marcum Anatomic Pathology Manager UAMS Little Rock AR 501-686-5941 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC on FNA
Linda I agree with you that is why I asked the question. We do not have a cryostat, so frozen sections are out of the question. I thought of making cytospin slides out of positive fluid but then you run into the problem of any positive cells making it onto the slide. Would any of you run the FNA slide without pretreatment and the FFPE control with the pretreatment? I appreciate everyone suggestions. Cindy Pyse, CLT, HT (ASCP) Histology Supervisor X-Cell Laboratories e-mail cp...@x-celllab.com -Original Message- From: Sebree Linda A [mailto:lseb...@uwhealth.org] Sent: Friday, April 09, 2010 12:40 PM To: Cynthia Pyse; Histonet Subject: RE: [Histonet] IHC on FNA This topic touches a nerve with me. We recently went back to receiving requests on FNAs after years of not doing any. When we were routinely staining them with IHC, we would use a frozen tissue section for a control as the same protocols generally worked for both, i.e. no HIER, protease, etc as may be needed for FFPE. When we stopped doing FNA IHC routinely, we eliminated our frozen section control inventory. Now that we're doing them again, it was decided to use FFPE controls and their accompanying IHC protocols. I do not agree with this practice as I feel there are too many differences between FNA preparations and FFPE sections. I was voted down so now we are using HIER, protease, etc. whatever the FFPE protocol calls for on these FNA preparations. Surprisingly, at least to me, there are still cells left on the FNA preps after even the harshest retrieval protocols. We also don't run negative controls of the FNAs. The argument being that there are totally different cells from one slide to the next so a comparison between a negative control slide and one that received antibody is useless. So that's what we're doing even though I don't agree with it. At the time this all came about, I also queried Histonet as to common practices so the archives should have some info. also. Linda A. Sebree University of Wisconsin Hospital Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Cynthia Pyse Sent: Friday, April 09, 2010 11:16 AM To: 'Histonet' Subject: [Histonet] IHC on FNA Happy Friday Histonetters What is everyone using for controls for FNA IHC stains. We routinely stain human FFPE sections and only have controls for tissue. Recently we had a few cases when the only sample was an FNA to perform IHC on. Any suggestions would be welcome . Regards Cindy Pyse, CLT, HT (ASCP) Histology Supervisor X-Cell Laboratories e-mail cp...@x-celllab.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] Re: Cd31 in pigs
Serotec's mouse anti-porcine CD31 (MCA1746) will work well in fresh, frozen, acetone-fixed pig tissues. It is very good for angiogenesis. It does not work with FFPE tissues. Liz Wilson Bethyl Labs -- Message: 4 Date: Fri, 9 Apr 2010 11:59:44 -0400 From: Jean-Martin Lapointe jm.lapoi...@accellab.com Subject: [Histonet] Cd31 in pigs To: histonet@lists.utsouthwestern.edu Message-ID: befd613bd39142499989f836556ddc8394a...@ace.accellab.lan Content-Type: text/plain; charset=iso-8859-1 Hi Joel, we have tried a few antibodies for CD31 in FFPE tissues in pigs and have had no luck. We currently use an antibody to vWF to stain pig endothelium. If someone has a working technique for CD31 in swine, I'd be happy to hear about it. Jean-Martin __ Jean-Martin Lapointe, DMV, MS, dACVP AccelLAB Inc Quibec, Canada J7H 1N8 jm.lapoi...@accellab.com Date: Fri, 9 Apr 2010 09:40:00 -0400 From: Joel Israel jo...@mcclainlab.com Does anyone have a procedure that WORKS for cd31 in pigs? I can't seem to get it to work no matter what I do. Any help will be greatly appreciated. Thanks.- joel ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] IHC controls for FNA's
It would depend on what type of tissue/ cells you are staining and what antibodies. Why not obtain some of the fresh tissue from your most commonly performed (and IHC stained) type of cases, or from tissues that you know would be positive for the antibodies in question and do an FNA on it yourself. Aspirate some cells and make smears and fix them as you do your FNA's. It may be a problem to do this contemporaneously but maybe storing some smears in 95% alcohol would be appropriate. Jeff Silverman ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
