[Histonet] RE: Endogenous Tau (normal Tau)
Hi Claudia, We use Dako anti-Human Tau (rabbit polyclonal) at a concentration of 1:8000. We do not use a pretreatment (HIER or protease). Staining is done on a Ventana stainer. Good luck, Willem Van: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] namens Segovia, Claudia [csego...@nsalabs.com] Verzonden: donderdag 6 november 2014 19:23 Aan: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] Endogenous Tau (normal Tau) Hi there, I am writing a poster on different Tau epitopes in human brain free floating sections. I have been unable to get any normal Tau and I wonder what could be the problem. Does anybody know what could be? Also, does anyone have a protocol that can share with me? Thank you! Claudia N. Segovia Senior Neurohistologist Antibody Specialist NeuroScience Associates 10915 Lake Ridge Drive Knoxville, TN 37934 865-675-2245 csego...@nsalabs.com STATEMENT OF CONFIDENTIALITY: The information contained in this electronic message and any attachments are intended for the exclusive use of the addressee(s) and may contain confidential or privileged information. If you are not the intended recipient, or the person responsible for delivering email to the intended recipient, be advised you have received this message in error and any use, dissemination, forwarding, printing or copying is strictly prohibited. Please notify NeuroScience Associates immediately at 865-675-2245 or at csego...@nsalabs.com, and destroy all copies of this message and any attachments. You will be reimbursed for reasonable costs incurred in notifying us. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Corpora amylacea are destroyed after antigen retrieval?
I am not sure if you are talking about the Beta Amyloid antibody, but if you are: the best way to do the antigen retrieval is 3 minutes in formic acid. Willem Van: histonet-boun...@lists.utsouthwestern.edu namens Pirici Daniel Verzonden: di 18-2-2014 23:59 Aan: Histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] Corpora amylacea are destroyed after antigen retrieval? Has anyone observed corpora amylacea on brain tissue being destroyed after antigen retrieval (for example microwaving in citrate buffer)? We are experiencing this problem, as we are interested to study these structures, and we are not sure if it is a common artifact when preparing slides for IHC... Thanks so much for your time! Daniel Pirici Nicolae Daniel, MD, PhD Department of Histology University of Medicine and Pharmacy Craiova Petru Rares Street 2, 200349 Craiova, Dolj Romania ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] M2D2 and SMAD
Hi, SMAD4 is very difficult on the Ventana platform. The only clone that would work on the Ventana to my knowledge(clone EP618Y, immunologic) has recently 'collapsed' (i hope that's the right word?) It means that it is not available anymore. Maybe there is some vendor that has a few vials left, but my supplier does not. If anybody knows of some other clone for the Ventana, I am very interested. MDM2: our sarcoma pathologist says that the MDM2 immuno is not reliable. Some liposarcomas will be negative. It would be better to do FISH, which is also very difficult because the fatty tissue often rinses off the slides. Willem Van: histonet-boun...@lists.utsouthwestern.edu namens Sue Hunter Verzonden: do 5-12-2013 16:50 Aan: Histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] M2D2 and SMAD Good morning Does anyone have suggestions for antibodies for M2D2 and SMAD? We have not been able to get these antibodies to work for us on the Ventana Ultras. If you do have suggestions, protocols and control tissue suggestions would also be appreciated. Thanks Sue Sue Hunter, Supervisor Advanced Diagnostics Beaumont Health System Royal Oak MI 248-898-5146 shun...@beaumont.edumailto:shun...@beaumont.edu ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Her2 Dual ISH and breast processing
Thank you for the references, I have found what I needed in this article: Minimum Formalin Fixation Time for Consistent Estrogen Receptor Immunohistochemical Staining of Invasive Breast Carcinoma. Goldstein NS, et. al., Am J Clin Pathol 2003;120:86-92 Willem My mistake: the web site is: http://www.histosearch.com/rene.html From: Rene J Buesa rjbu...@yahoo.com To: Hoekert, W.E.J. w.e.j.hoek...@olvg.nl; vtol...@cox.net vtol...@cox.net; histonet@lists.utsouthwestern.edu [] Willem, yes, there are. Look through the attached references which include some for breast fixation. There have been some more recently as well, just go on scholar.google.com . Tim Morken Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Centero Medical Center Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Her2 Dual ISH and breast processing
I was wondering,is there any literature on this subject? i.e. the minimal required fixation time of breast tissue in order to get reliable immuno staining (Oestro, Prog and Her2neu and ISH). We immunologists are alway trying to convince our pathologists about the importance of good fixation, but the pathologist are always in a hurry to get the diagnosis out. Many mamma biopsies do not get to be fixed long enough (some are even put in the processor after a few hours of fixation). It would be helpfull if I can show them some literature to back me up. Willem Van: histonet-boun...@lists.utsouthwestern.edu namens Rene J Buesa Verzonden: ma 10-12-2012 18:23 Aan: vtol...@cox.net; histonet@lists.utsouthwestern.edu Onderwerp: Re: [Histonet] Her2 Dual ISH and breast processing Val: Acknowledging that you have a fixation problem is the fundamental step to solving your staining problems. You state that the slices have now a consistent thickness of between 2-3 mm and that is great BUT what about the fixation time? Thin slices is a first step but until you have the slices properly fixed, you will keep having some problems. Even when one should never assume, I assume that you are using NBF at room temperature. With that fixative and under those temperature conditions, your 2-3 mm thick breast slices will require 4 hours to be fully penetrated; will require 24 hours to be 100% covalent bound and will require 96 hours to be completely cross-linked. The 48 hours of maximum exposition to NBF recommended by ASCO-CAP will guarantee a 100% cross-linking of 1 mm thick slices, but your 2-3 mm slices will require 96 hours to be completely cross-linked. Until you reach an optimum fixation you will keep having sporadic problems of your protocols (depending on greater or lower fat contents of the samples). René J. From: vtol...@cox.net vtol...@cox.net To: histonet@lists.utsouthwestern.edu Sent: Monday, December 10, 2012 11:48 AM Subject: [Histonet] Her2 Dual ISH and breast processing Hello all-- My lab is having some problems with inconstant results with our Her2 Dual ISH on the Ventana Ultra. Right now, we know we have a problem with our large breast tissue being under-fixed. There have been many gripes to the PAs and residents about the thickness of the tissue sections and they have listened. We are now getting breast sections that are consistently cut between 2-3mm in thickness. However, we are still having issues with inconsistency in our dual ISH staining. Many times, the staining is absent. We are currently processing our breast tissue on a Tissue-Tek VIP6 processor. Are there any labs out there that are using this processor and also running Her2 Dual ISH on the Ventana with nice, consistent results? If so, would any of you be willing to share your processing protocol? Thanks in advance for your help! Val ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] MSI antibodies
Hi Kenia I guess you will need at least two blocks because they are never 'positive' all four of them in the same tumour. You might search your archive for young people with colon cancer. There is a good chance that you will find positive material. Willem Hoekert OLVG The Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens lynch kenia Verzonden: wo 28-11-2012 16:09 Aan: Histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] MSI antibodies Need help please..does anyone have a colon block, or would be will ing to cut me a few slides of colon, that has stained positive (which is actually no staining) for the MSI antibodies; MLH1,PMS2, MSH6, MSH2 ? Thanks! Kenia Lynch, M.T./H.T. Histology/Molecular Supervisor Caldwell Memorial Hospital 321 Mulberry Street SW Lenoir, NC 28645 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Pronase antigen retrieval
Hello Eva, We are not using pronase anymore, but when we did, we were doing it at room temperature. (15 mg in 250 ml PBS, sigma P-5147). It worked fine. Our Trypsin antigen retrieval, we were doing it at 37C as follows: the evening before the retrieval, we put some PBS in the 37C stove, the next morning we dissolved the trypsin in the warm PBS and we put the slides in it (again in the stove, for 15 minutes). Good luck, Willem Van: histonet-boun...@lists.utsouthwestern.edu namens Eva Permaul Verzonden: di 11-9-2012 19:36 Aan: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] Pronase antigen retrieval Hello out there in Histoland, I have to do some bulk runs with Pronase antigen retrieval. Up until now the number of slides have been so few that I have been able to do the antigen retrieval by dropping pronase on each slide and incubating them in a humidified chamber at 37C for 20min. I now have so many slides that my manager has suggested that I make up large amounts of the Pronase (250ml) and submerge the slides. She suggested placing the container in the oven earlier to allow the Pronase to get to 37C before I add the slides. How long is the Pronase ok to be keep at 37C before I use it? How long in advance can I place it in the 37C before it looses its ability to be used as an antigen retrieval? Thanks for all your help, Eva Permaul HTSR ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] How to make HP control tissue?
Hi Histonetters, Does anybody has experience in making your own HP control tissue? I am tired of using positive biopsies of patients since they are always almost finished. I have heard of a procedure on making HP controls but I am not exactly sure how it is done. I have tried the following: I went to the microbiology department and asked for some freshly grown HP bacteria. We scraped them of the petridishes and put them in formalin. I had them fixed for 24 hours and then I injected them into some already fixed lung tissue and processed it as normal. But if I do a HP stain on that tissue, I don't see any bacteria. Probably they rinse off during the process. Can anyone tell me how to do it? Thanks in advance, Willem Hoekert OLVG The Netherlands Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] MDM-2 on Ventana, which clone?
Hi Histonetters, I am having a very hard time in setting up the MDM-2 antibody on our Ventana Benchmark XT's. I am using clone SMP14 from Becton Dickinson, which was working fine on the Labvision. I have tried everything (even incubating overnight at 4ºC) and I am out of idea's. Maybe I should use another clone, for instance 1B10 from Monosan? Can somebody please give me a good protocol? Thank's in advance, Willem Hoekert Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] SMAD4
Which stainer are you using? Because some clones will not work on a Venatana stainer. Willem Van: histonet-boun...@lists.utsouthwestern.edu namens Chakib Boussahmain Verzonden: ma 23-4-2012 4:20 Aan: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] SMAD4 Hello histonet, Does anyone uses SMAD4 antibody? If so, can you tell where can I find a good one? Can you also share the staining protocol? dilution? epitope retrieval? Your help will be appreciated so much. Thank you. Chakib Boussahmain Histotech ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] IHC and negative controls?
We are not using negative controls. We only add positive controls at the bottom of our patient slides. Sometimes there are 3 different types of control tissue (a TMA), so that you can use the same control block for several antibodies. You could add a negative control in the same block as the positive control and add these at the bottom of your slides (or at the top, if you wish). Willem Hoekert OLVG The Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens Tom McNemar Verzonden: do 16-2-2012 16:27 Aan: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] IHC and negative controls? Hi All, We have one Ventana XT and often exceed the slide capacity. Our pathologists no longer want us to run negatives and will use internal negatives instead. Is this common practice? Thanks. I appreciate any feedback. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcne...@lmhealth.orgmailto:tmcne...@lmhealth.org www.LMHealth.orgfile:///C:\Documents%20and%20Settings\TMCNEMAR\Application%20Data\Microsoft\Signatures\www.LMHealth.org file:///C:/Documents%20and%20Settings/TMCNEMAR/Application%20Data/Microsoft/Signatures/www.LMHealth.org This e-mail, including attachments, is intended for the sole use of the individual and/or entity to whom it is addressed, and contains information from Licking Memorial Health Systems which is confidential or privileged. If you are not the intended recipient, nor authorized to receive for the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this e-mail and attachments is prohibited. If you have received this in error, please advise the sender by reply e-mail and delete the message immediately. You may also contact the LMH Process Improvement Center at 740-348-4641. E-mail transmissions cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. The sender therefore does not accept liability for any errors or omissions in the contents of this message, which arise as a result of e-mail transmission. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] HSV1 and HSV2
For the immunologic pAB cocktail we are using Protease 1 (4 min) and ab incubation of 32 min. The only problem is that mast cells are also staining, it is a side effect of the protease 1. (AB concentration 1:400) Willem Hoekert OLVG Amsterdam The Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens Evans, Andria B Verzonden: vr 23-12-2011 18:48 Aan: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] HSV1 and HSV2 I am currently having issues with our HSV1 and HSV2 staining tissue components that it shouldn't be/background. We are running Ventana Benchmark XTs and Ultras. Using Dako's polyclonal rabbit concentrate. Does anyone out there in histoland have a concentration with a protocol that works and looks clean? Thank you for your help! Andria B Evans HTL(ASCP)CM This email was sent securely from the LGHealth Email Service 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] GLUT-1 antibody
Hi Sandra, I am using Neomarkers (Glut 1 Ab-1, polyclonal) on the Benchmark XT. 30' cc1 32' ab incubation 1:50 It works fine. Willem Hoekert OLVG Amsterdam The Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens Diaz, Sandra Verzonden: za 26-11-2011 0:38 Aan: 'histonet@lists.utsouthwestern.edu' Onderwerp: [Histonet] GLUT-1 antibody We recently purchased Cell Marque's Glut-1 antibody and it is not staining correctly for us. Does anyone have a working protocol for the Benchmark Ultra? Or is the antibody from another company work anyone? Sandra Diaz H.T. IHC Department, HMFW 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 Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Fatty mamma tissue keeps on washing off
Hi histonetters Here is an update on my struggle to keep the fatty mamma tissue on the slides. I got a few responses on my mail so I have tried a few other things. Post fix the slides. Deparaffinize the slides using xylene and hydrate to water, post fix the slides in 10% NBF for 10 minutes (I did it for about 4 hours) and proceed as usual. That worked for a few of my hopeless cases. Still, some other cases would still come of the slides. I have also tried the Dako slides. Indeed they work great. Of the 4 cases that washed off no matter what, all but one stayed on the Dako slides. So from now on, if we have a case that washes off, we will be using the Dako slides. Thanks for your help, Willem Hoekert OLVG AMsterdam The Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens Hoekert, W.E.J. Verzonden: do 20-10-2011 12:09 Aan: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] Fatty mamma tissue keeps on washing off Hi Histonetters, Every once in a while, we have mamma tissue that washes off the slides completely. The reason for this is that the tissue is still too fat and/or not sufficiently fixed. There is no problem with the HE stain, but the immuno will wash off. What do you do in such cases? I have tried the following: 1: Baking the slides for about 3/4 hours on superfrost slides (that works sometimes but not always) 2: Wood glue slides (that works sometimes but not always) 3:We put the blocks back in hot parraffin for several hours (that usually works) but I was told that this will have a bad influence on the Her2neu stain: a 2+ could become a 1+. I know, the solution is to be sure that the tissue is fixed long enough and that the fat has been removed sufficiently using acetone, but sometimes that is just not the case. How do you cope with this? Willem Hoekert PA-lab OLVG Amsterdam The Netherlands Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Fatty mamma tissue keeps on washing off
Hi Histonetters, Every once in a while, we have mamma tissue that washes off the slides completely. The reason for this is that the tissue is still too fat and/or not sufficiently fixed. There is no problem with the HE stain, but the immuno will wash off. What do you do in such cases? I have tried the following: 1: Baking the slides for about 3/4 hours on superfrost slides (that works sometimes but not always) 2: Wood glue slides (that works sometimes but not always) 3:We put the blocks back in hot parraffin for several hours (that usually works) but I was told that this will have a bad influence on the Her2neu stain: a 2+ could become a 1+. I know, the solution is to be sure that the tissue is fixed long enough and that the fat has been removed sufficiently using acetone, but sometimes that is just not the case. How do you cope with this? Willem Hoekert PA-lab OLVG Amsterdam The Netherlands Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] P 16
Hi Phyllis, We are doing a double stain (Mib-1 - P16) on cervix biopsies in which the P16 antibody is coloured with AP. It works fine, our pathologists are quite fond of it. Almost every day we have a few slides. Willem Hoekert OLVG, Amsterdam The Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens Phyllis Thaxton Verzonden: wo 28-9-2011 21:14 Aan: Histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] P 16 Has anyone ever used P16 with AP Red detection on the Ventana Benchmark? Phyllis Jordan HT(ASCP)QIHC DCH Regional Medical Center Tuscaloosa, AL ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Microsatellite Instability Antibodies
We are currently running the Ventana IHC stainers with the UltraView Detection. Just like us! MLH-1: Cell Marque (ref 760-4264). 30' cc1, 32' ab incubation. MSH-2: Cell Marque (ref 760-4265). 60' cc1, 60' ab inc. PMS2: Becton Dickinson, clone A16-4, cat nr 556415. 30' cc1, 32' ab inc. +amplification MSH6: Becton Dickinson, clone 44, cat nr 610919. 60' cc2, 60' ab inc. MSH-6 does not give very nice results, however, it works. Good Luck, Willem Van: histonet-boun...@lists.utsouthwestern.edu namens Evans, Andria B Verzonden: do 24-3-2011 13:31 Aan: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] Microsatellite Instability Antibodies I am currently working up the MSI Antibodies (MLH1, MSH6, MSH2, and PMS2) and need some input on the protocol that you may be running in your lab and what clone(manufacture) you are using. We are currently running the Ventana IHC stainers with the UltraView Detection. Thanks for your help! Andria B Evans HTL(ASCP)CM This email was sent securely from the LGHealth Email Service 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. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] RE: Procedure for making Gram Control
OK! I always thought that a Slim Jim was some kind of candy, but it is a snack sausage. Here in The Netherlands we have Bi-Fi. I will try one of those. Thanks, Willem Hoekert Amsterdam, The Netherlands. Van: histonet-boun...@lists.utsouthwestern.edu namens Thomas Jasper Verzonden: do 17-3-2011 18:32 Aan: Hoekert, W.E.J. CC: histonet@lists.utsouthwestern.edu Onderwerp: RE: [Histonet] RE: Procedure for making Gram Control Some sort of small, snack sausage of questionable quality. I'm not familiar with anything like that in Europe, but maybe you could determine that somehow. Good Luck! Tom Jasper Bend, Oregon -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Hoekert, W.E.J. Sent: Thursday, March 17, 2011 1:47 AM To: Walter Benton; Hayes, Randi (HorizonNB); histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Procedure for making Gram Control What would be the equivalent of a Slim Jim in Europe? The Netherlands to be more precise? Van: histonet-boun...@lists.utsouthwestern.edu namens Walter Benton Verzonden: wo 16-3-2011 15:07 Aan: Hayes, Randi (HorizonNB); histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] RE: Procedure for making Gram Control Making you own...not sure how to do that, but Slim Jim's work in a pinch. Walter Benton HT(ASCP)QIHC Histology Supervisor Chesapeake Urology Associates 806 Landmark Drive, Suite 126 (All Deliveries to Suite 127) Glen Burnie, MD 21061 443-471-5850 (Direct) 410-768-5961 (Lab) 410-768-5965 (Fax) wben...@cua.md From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Hayes, Randi (HorizonNB) [randi.ha...@horizonnb.ca] Sent: Wednesday, March 16, 2011 10:03 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Procedure for making Gram Control Hello out there in Histoland, I'm looking for a procedure for making your own Gram controls. Any assistance would be appreciated. Thanks, Randi /pre--- Horizon Health Network Disclaimer ---brbrThis e-mail communication (including any or all attachments) is intendedbronly for the use of the person or entity to which it is addressed and maybrcontain confidential and/or privileged material. If you are not the intendedbrrecipient of this e-mail, any use, review, retransmission, distribution,brdissemination, copying, printing, or other use of, or taking of any action inbrreliance upon this e-mail, is strictly prohibited. If you have received thisbre-mail in error, please contact the sender and delete the original and anybrcopy of this e-mail and any printout thereof, immediately. Yourbrco-operation is appreciated.brbrLe présent courriel (y compris toute pièce jointe) s'adresse uniquement àbrson destinataire, qu'il soit une personne ou un organisme, et pourraitbrcomporter des renseignements privilégiés ou confidentiels. Si vous n'êtesbrpas le destinataire du courriel, il est interdit d'utiliser, de revoir, debrretransmettre, de distribuer, de disséminer, de copier ou d'imprimer cebrcourriel, d'agir en vous y fiant ou de vous en servir de toute autre façon.brSi vous avez reçu le présent courriel par erreur, prière de communiquerbravec l'expéditeur et d'éliminer l'original du courriel, ainsi que toute copiebrélectronique ou imprimée de celui-ci, immédiatement. Nous sommesbrreconnaissants de votre collaboration.pre CONFIDENTIALITY NOTICE: The information contained in this electronic message is intended solely for the personal and confidential use of the designated recipient(s) named above and may contain information that is protected from disclosure under applicable law. If you are not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this transmission is strictly prohibited. If you have received this transmission in error, please notify the transmitting person/department immediately by email or telephone (410) 581-5881 and delete the message without making a copy. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] RE: Procedure for making Gram Control
What would be the equivalent of a Slim Jim in Europe? The Netherlands to be more precise? Van: histonet-boun...@lists.utsouthwestern.edu namens Walter Benton Verzonden: wo 16-3-2011 15:07 Aan: Hayes, Randi (HorizonNB); histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] RE: Procedure for making Gram Control Making you own...not sure how to do that, but Slim Jim's work in a pinch. Walter Benton HT(ASCP)QIHC Histology Supervisor Chesapeake Urology Associates 806 Landmark Drive, Suite 126 (All Deliveries to Suite 127) Glen Burnie, MD 21061 443-471-5850 (Direct) 410-768-5961 (Lab) 410-768-5965 (Fax) wben...@cua.md From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Hayes, Randi (HorizonNB) [randi.ha...@horizonnb.ca] Sent: Wednesday, March 16, 2011 10:03 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Procedure for making Gram Control Hello out there in Histoland, I'm looking for a procedure for making your own Gram controls. Any assistance would be appreciated. Thanks, Randi /pre--- Horizon Health Network Disclaimer ---brbrThis e-mail communication (including any or all attachments) is intendedbronly for the use of the person or entity to which it is addressed and maybrcontain confidential and/or privileged material. If you are not the intendedbrrecipient of this e-mail, any use, review, retransmission, distribution,brdissemination, copying, printing, or other use of, or taking of any action inbrreliance upon this e-mail, is strictly prohibited. If you have received thisbre-mail in error, please contact the sender and delete the original and anybrcopy of this e-mail and any printout thereof, immediately. Yourbrco-operation is appreciated.brbrLe présent courriel (y compris toute pièce jointe) s'adresse uniquement àbrson destinataire, qu'il soit une personne ou un organisme, et pourraitbrcomporter des renseignements privilégiés ou confidentiels. Si vous n'êtesbrpas le destinataire du courriel, il est interdit d'utiliser, de revoir, debrretransmettre, de distribuer, de disséminer, de copier ou d'imprimer cebrcourriel, d'agir en vous y fiant ou de vous en servir de toute autre façon.brSi vous avez reçu le présent courriel par erreur, prière de communiquerbravec l'expéditeur et d'éliminer l'original du courriel, ainsi que toute copiebrélectronique ou imprimée de celui-ci, immédiatement. Nous sommesbrreconnaissants de votre collaboration.pre CONFIDENTIALITY NOTICE: The information contained in this electronic message is intended solely for the personal and confidential use of the designated recipient(s) named above and may contain information that is protected from disclosure under applicable law. If you are not the intended recipient, or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this transmission is strictly prohibited. If you have received this transmission in error, please notify the transmitting person/department immediately by email or telephone (410) 581-5881 and delete the message without making a copy. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] ER staining in Tonsil.
Yes, we see the same. We also see staining in follicles in appendix and colon. Van: histonet-boun...@lists.utsouthwestern.edu namens Andrea Verzonden: do 9-12-2010 15:05 Aan: Histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] ER staining in Tonsil. Good Morning, Is it usual to get low ER staining in tonsil? ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Formalin
Not here in The Netherlands. Van: histonet-boun...@lists.utsouthwestern.edu namens Laurie Colbert Verzonden: vr 12/11/2010 15:42 Aan: godsgal...@aol.com; Histonet@lists.utsouthwestern.edu Onderwerp: RE: [Histonet] Formalin Not us - in Pasadena CA. -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of godsgal...@aol.com Sent: Thursday, November 11, 2010 2:45 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Formalin Ok, so how many of you are allowed to dump formalin down the drain? Sent from my Verizon Wireless Phone ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] PMS2
We used the same antibody but then at 1:1200, retrieved with PH9 (PT module buffer 4, thermo) on a labvision stainer. We had beautiful results. Now, we switched to Ventana Benchmark XT (1:50, 30' cc1 + amplification) and our results are quite poor for PMS2: many cytoplasmatic staining. So if anybody has a good protocol for the Benchmark XT, I am very interested. Regards, Willem Hoekert OLVG, Amsterdam Van: histonet-boun...@lists.utsouthwestern.edu namens Dana Settembre Verzonden: wo 25-8-2010 12:12 Aan: dianar...@aol.com; histonet@lists.utsouthwestern.edu Onderwerp: Re: [Histonet] PMS2 Diane, When I was doing PMS2 I was using BD's mouse antibody @ 1:10 Retreived with Dako's TRS in a steamer for 40 min, incubated the antibody for 60 minutes and I used a labelled polymer for the detection (Dako's Envision + Mouse) It was difficult to work up. Good Luck, Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJUSA dianar...@aol.com 08/24/10 8:47 PM Can anyone share their protocol for PMS2? I just keep getting background staining. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Smad4 on a Ventana Benchmark XT
Hi Histonetters, We are thinking of taking the Ventana Benchmark XT, and I am testing it right now. So far, it is impossible to get the Smad4 up and running. I have tried everything: -Pretreatment with cc1 and cc2 for up to 90 minutes -Incubation of primairy AB for up to 90 minutes -Incubation at 37 degrees and room temperature -Pretreatment using a PT module, and even with a microwave -Increasing the concentration of AB But nothing works. Maybe I should try another clone? Right now we are using clone B-8 from Santa Cruz, and we are using HIER (EDTA) as pretreatment. Does anyone have any suggestions? I hope to hear from you. Willem Hoekert OLVG, PA Lab The Netherlands Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] How to do an EBER on cytological material?
Dear histonetters, I was wondering if it is possible to do an EBER ISH on a cell-smear. What pretreatment should I use? 1: Fix the cells in formalin (let's say overnight, or else one or two hours), and than use the proteinase K digestion step. 2: Or maybe it is enough to fix the cells in cold acetone, and skip the proteinase K step? Or will the RNA leak out of the cells? Does anybody has experience whit this? We only have two slides left, and we don't have (cytological) control material, only FFPE control material. Also, I think that we should bake the slides prior to the asessment because we might lose a lot of cells during the (stringent) washing steps. Maybe it is not possible at all because the RNA is broken down allready (the cell smears were taken 3 days ago, they were stored at room temperature). I hope you folks can help me out. Willem Hoekert OLVG, The Netherlands Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] [ SOLVED ][ Histonet ] Strange circles in IHC slides
Are you sure that you don't introduce air bubbles when you put your slides into the coverplates? The antibody will not touch the tissue if there is an air bubble. Willem Hoekert Van: histonet-boun...@lists.utsouthwestern.edu namens Rene J Buesa Verzonden: do 25-2-2010 16:42 Aan: histonet@lists.utsouthwestern.edu; histonet.nos...@vneubert.com Onderwerp: Re: [Histonet] [ SOLVED ][ Histonet ] Strange circles in IHC slides To me it seems that the sections after being picked from the water bath were not completely drained and the dewaxing process was incomplete in a way that the round areas kept certain amount of paraffin wax that prevented the reagents reactions. The fact that the areas are round are an indication that water was involved, since water always leave a round imprint, due to its surface tension. I would suggest that you dewax the sections with a 2% aq. solution of dish washer soap. Dewaxing with xylene sections containing water will be incomplete because it does not mix completely with water but the detergent will mix with the water and will better remove the paraffin.René J. --- On Thu, 2/25/10, histonet.nos...@vneubert.com histonet.nos...@vneubert.com wrote: From: histonet.nos...@vneubert.com histonet.nos...@vneubert.com Subject:[Histonet] [ SOLVED ][ Histonet ] Strange circles in IHC slides To: histonet@lists.utsouthwestern.edu Date: Thursday, February 25, 2010, 8:34 AM Hello Histonet, it has been a while (~10 months) since I posted a problem about uneven immuno-staining with specimen showing unstained circles after manual staining with HRP-polymere/DAB method; complete mail see below, response mails see Histonet archive (via website). Links to pictures I took: http://img12.imageshack.us/img12/8513/ts0402162049.jpg http://img13.imageshack.us/img13/6514/ts0402162104.jpg The problem occured suddenly, without having changed any reagents or methods. Things I changed to avoid the unstained spots: *Adding 0,05% Tween 20 to TBS *Blocking peroxidase in coplin jar, not mounted in racks *Lowering antibody concentration which temporarily produced better results. After reviewing a big number of slides it showed up that most of the tissue affected was lung, liver and kidney which mostly means a lot of blood in the tissue when fixation in formalin starts. Erythrocytes, granulocytes and macrophages show a lot of endogenous and pseudoendogenous peroxidase activity. This is how it's done since then: Slides are taken from racks into a big coplin jar with 3% H2O2 diluted in distilled water (demineralized H2O). A slow magnetic stirrer on the bottom of the jar keeps the solution floating around the tissue, removing any O2 bubble that might appear. Slides then are remounted and rinsed a lot with TBS-T. Thank you for all your help, though it's a little late... V. Neubert, Germany - Original Message - From: histonet.nos...@vneubert.com To: histonet@lists.utsouthwestern.edu Date: 02.04.2009 18:12:18 Subject: [Histonet] Strange circles in IHC slides [...] http://img12.imageshack.us/img12/8513/ts0402162049.jpg http://img13.imageshack.us/img13/6514/ts0402162104.jpg [...] So, has ever anyone experienced sth. like this? My conjugate control (every step except the antibody) was fine, nothing to be seen about DAB and no circles at all. I used Shandon single-use coverplates, sterile buffer, fresh antibody aliquots. Any idea? Thanks, V. Neubert ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] CISH - HER2
We are using Spot Light her2 Cish from invitrogen (manual, FDA approved). Willem Hoekert Pathology OLVG, Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens Taylor, Jean Verzonden: di 5-1-2010 22:02 Aan: 'histonet@lists.utsouthwestern.edu'; 'ih...@googlegroups.com' Onderwerp: [Histonet] CISH - HER2 The pathologists that I work for are requesting that our lab look into starting up CISH mainly for HER2. What company are labs purchasing their CISH HER2 kits from? Does anyone use Dako's DuoCISH kit along with their HER2 FISH pharmDx kit? We currently run the old Dako Autostainers, but are in the market for purchasing a new IHC stainer as well. Thanks for your help! Jean Taylor, HT(ASCP)QIHC IHC Tech Meriter Health Services Madison, WI ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] To my fellow histotechs....
* Cyclin D1 : NeoMarkers, SP4 * CD56 : NeoMarkers, 123C3.D5 * CD138 : Dako, M115 * Kappa : Dako, A0191 (Polyclonal) * Lambda : Dako, A0193 (Polyclonal) * MUM-1 : Dako, MUM1p Willem Hoekert Pathology, OLVG The Netherlands Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Labvision immunostainers skip slides
Thank you all for your replies, they are of great use to me. I now understand how the skipped staining can occur. I have told our support guy to look at the reagent sensor system, and he will do so today. He told me that he has already replaced them a while ago, and that the new ones are relatively stable. However, he also told me that a few weeks ago, they changed the settings of the probe, so that the probe is now descending to the bottom of the vial when it is sucking up the reagents. So actually, I still don't understand how we can get the skipped staining. I am afraid that this problem will not be fixed very easy, if it can be fixed at all. I understand that Labvision is working on this problem and that in october next year they hope to have a solution. We have decided to deparaffinize with xylene since it seems that the PT modules are not effective enough in doing so. We will keep a closer eye on them as some of you have pointed out that they can be a cause of problems. We also have irregular staining, but this occurs also with slides that have not been treated in the PT modules. PS: we use Tris Hcl buffer (10 times concentrated) from Klinipath, which contains tween already. And yes, we do get good support, but still the problems are not solved. Thanks again, Willem Hoekert Pathology Lab, OLVG Amsterdam The Netherlands Van: histonet-boun...@lists.utsouthwestern.edu namens Hoekert, W.E.J. Verzonden: vr 9-10-2009 13:19 Aan: histonet@lists.utsouthwestern.edu Onderwerp: [Histonet] Labvision immunostainers skip slides Dear Histonetters, A few months ago, we bought 2 Labvision Autostainers 720 from Thermo (immunostainers). We have some bad experiences with them. They seem to skip some slides every now and than (it happens maybe once every 3-4 weeks, we stain about 150 - 200 slides a day). If so, our positive controls are virtually completely negative, and the patient slides are negative as well. If this happens, we see empty 'holes' in the nuclei. Click here to see an image: http://www.immunoportal.com/modules.php?name=gallery2g2_itemId=14523g2_imageViewsIndex=1 All the other antibodies from the same run work fine, and if we repeat the failed cases (without changing anything) they turn out fine. Does anybody recognize this problem or knows what is going on? We dewax in PT modules (the problem occurs both with citraclere and with PT Module Buffer 1 from immunologic). I hope to hear from you. Willem Hoekert Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Labvision immunostainers skip slides
Dear Histonetters, A few months ago, we bought 2 Labvision Autostainers 720 from Thermo (immunostainers). We have some bad experiences with them. They seem to skip some slides every now and than (it happens maybe once every 3-4 weeks, we stain about 150 - 200 slides a day). If so, our positive controls are virtually completely negative, and the patient slides are negative as well. If this happens, we see empty 'holes' in the nuclei. Click here to see an image: http://www.immunoportal.com/modules.php?name=gallery2g2_itemId=14523g2_imageViewsIndex=1 All the other antibodies from the same run work fine, and if we repeat the failed cases (without changing anything) they turn out fine. Does anybody recognize this problem or knows what is going on? We dewax in PT modules (the problem occurs both with citraclere and with PT Module Buffer 1 from immunologic). I hope to hear from you. Willem Hoekert Disclaimer: Dit e-mail bericht is uitsluitend bestemd voor de geadresseerde(n). Verstrekking aan en gebruik door anderen dan geadresseerden is niet toegestaan. Indien u niet de geadresseerde bent, wordt u verzocht de verzender hiervan op de hoogte te stellen en het bericht te verwijderen. In verband met electronische verzending kunnen aan dit e-mail bericht geen rechten worden ontleend. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
