[Histonet] Frozens and antigen retrieval
Hi all, A question from my student: antigen retrieval when using frozens for immunofluorescence -yes or no? The protocol suggested by the antibody company lists the retrieval as an option. I am used to do retrieval on paraffin sections, but not on cryo sections. Hit me with your opinions on this :-) Thanks in advance, Daniela Sent from my iPad __ This email has been scanned by the Symantec Email Security.cloud service. For more information please visit http://www.symanteccloud.com __ ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] Mohs
I assume you are talking about HT(ASCP) Registered techs. The majority of Mohs Techs are off-the-street people. Here in FL for Registered Licensed techs, between $25 - $30 / hour. The off-the-street folks $14 - $18, sometimes as low as $12.00 / hr. Yours, Dave From: Rebecca a. Johnson r...@bluemarble.net To: histonet Histonet@lists.utsouthwestern.edu Sent: Monday, July 2, 2012 6:37 PM Subject: [Histonet] Mohs Need to know what Mohs techs are getting paid. Thanks Becky ___ Histonet mailing list Histonet@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] Frozens and antigen retrieval
Usually not. Fixation cross-links the proteins, which can mask the epitope (=antibody binding site of the antigen). So antigen retrieval breaks the fixative cross-links, exposing the epitope. If there's no fixation, there's no cross-links, so the epitope is usually exposed and available to easily bind to the antibody. Plus, there's no destruction of tissue morphology if you're not using antigen retrieval, so the quality of the section looks much nicer. That being said, there may be some antibody out there that still needs antigen retrieval on frozen section, but then the company's protocol probably wouldn't say optional. So try it the first time without antigen retrieval. Peggy A. Wenk, HTL(ASCP)SLS Schools of Histotechnology William Beaumont Hospital Royal Oak, MI 48073 The opinions expressed are mine, and do not reflect Beaumont. -Original Message- From: Daniela Bodemer Sent: Wednesday, July 04, 2012 7:02 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Frozens and antigen retrieval Hi all, A question from my student: antigen retrieval when using frozens for immunofluorescence -yes or no? The protocol suggested by the antibody company lists the retrieval as an option. I am used to do retrieval on paraffin sections, but not on cryo sections. Hit me with your opinions on this :-) Thanks in advance, Daniela Sent from my iPad __ This email has been scanned by the Symantec Email Security.cloud service. For more information please visit http://www.symanteccloud.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] Immuno Controls
Looking for the same Ian -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Carlos Hernandez Sent: Tuesday, July 03, 2012 11:34 PM To: Histonet Subject: [Histonet] Immuno Controls Hi All! I was wondering if there is anybody out there that has an over abundance of controls for mart/melan a, s100, hmb45, pan keratin, ki-67, and mitf that they are will to share or sell? Please let me know if you can help me out. Thanks, Carlos ___ Histonet mailing list Histonet@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] 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
Re: [Histonet] How to make HP control tissue?
It may be hard to find someone to help you in the States. Doing that is against regulations. We must use human tissue with a natural infection I we are going to be using it as a control for human medical testing. That being said, you are right, your hp is washing out of the tissue. My suggestion is to not fix the hp culture, inject it fresh into unfixed tissue. Preferable fresh autopsy stomach or intestine. Allow it to culture (and attach) for about an hour or 2 and then fix. Be careful too long out of fixative and autolysis will occur. I hope that works. I have no experience with lung. It should work, but I do not know if the environment would cause the bacteria to attach and grow. Will Sent from my iPhone On Jul 4, 2012, at 11:09 AM, Hoekert, W.E.J. w.e.j.hoek...@olvg.nl wrote: 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] How to make HP control tissue?
Hi Willem, H. pylori needs the low pH of the stomach to survive and grow. It won't naturally be in lung tissue. When looking for HP, it's found on the mucosa and gastric pits of the stomach. Since you're looking for it in a specific place, I think putting it in lung is a bad idea. Your best bet for control tissue is a positive gastrectomy specimen. You could chop that up into a thousand pieces and never run out. We've had several cases like this. Send me your address and a FedEx account number and I'll see about send you a block or two. Mark On Wednesday, July 4, 2012, Hoekert, W.E.J. wrote: 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 mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] PCNA sc-7907?
Hello Histonet - Has anyone had any success staining with PCNA sc-7907 on frozen, paraformaldehyde-fixed tissue? I am experiencing a lot of background, and so far diluting down to 1:5000 hasn't helped at all. Any advice would be very much appreciated! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] PCNA sc-7907?
dilute antibody with %1 BSA.. and use %1 BSA for serum blocking... On Wed, Jul 4, 2012 at 10:59 PM, Mehmet Fatih BOZKURT fbozk...@gmail.comwrote: dilute antibody with %1 BSA.. and use %1 BSA for serum blocking... On Wed, Jul 4, 2012 at 10:21 PM, E Wolfe ewolfe...@gmail.com wrote: Hello Histonet - Has anyone had any success staining with PCNA sc-7907 on frozen, paraformaldehyde-fixed tissue? I am experiencing a lot of background, and so far diluting down to 1:5000 hasn't helped at all. Any advice would be very much appreciated! ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Mehmet Fatih BOZKURT, DVM, PhD Afyon Kocatepe University Faculty of Veterinary Medicine Department of Pathology 03030, ANS Campus Afyonkarahisar-TURKEY Tel: +902722281312-173/237 -- Mehmet Fatih BOZKURT, DVM, PhD Afyon Kocatepe University Faculty of Veterinary Medicine Department of Pathology 03030, ANS Campus Afyonkarahisar-TURKEY Tel: +902722281312-173/237 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] (no subject)
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