[Histonet] indian ink- thank you
Dear everyone, I just wanted to thank you for all the tips concerning the use of indian ink. They helped a lot! Vielen Dank, Frauke ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] indian ink
Dear everyone, when I was working in pathology gross section lab, we used black dye (and blue, green etc,Tusche in german) for orientation of the gross section specimens. We fixed the dye with Bouin's solution. Now I was asked for this dye, but I don't know if this was the conventional india ink you can buy for drawing in paper shops or was this a special dye. I don't found anything in commen pathology supplier catalogs and can't reach anyone of my former colleagues who might know. Can you help me out of this confusion?! Thanks a lot (Vielen Dank) Frauke ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] golgi stain- please help
Dear Histonetters, I was performing a golgi stain on mouse brains and my treated animal group gave nice results but the untreated control group did not show any staining at all (it looks like the staining didn't work). Does anyone know / have an idea, if I could repeat the staining on these vibratome sections? I read something about deimpregnation of golgi stained slides, but found no protocol and I'm not sure if I can perform another fresh golgi stain with these slides. Any suggestion is welcome!!! Thanks to all of you, Frauke ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] myelin basic protein
Hi everbody, I'm currently looking for an antibody that stains myelin in mouse brains (no animals with EAE). Does anyone have experences with antibodies against MBP, MAG or other myelin proteins in mouse brains and can give me a suggestion, which one is the best to order for paraffin embedded tissue? Thanks a lot Frauke ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: Re: AW: [Histonet] Chemicals that inactivate the primary antibody
Hi TF, we do IF with two 1.ABs out of mice using a sequential protocol from abcam: www.abcam.com/technical We perform a second blocking step with serum (higher conc. than the first one) after the first Fluorescence was added, do not perform another retrieval procedure (my ABs need the same) and perform the following incubation steps in the dark (we switch the light off and cover the slides after adding the ABs) to prevent fading. Until now, I was sure, this procedure works but maybe the two antigens don't colocalize but it is just a staining artifact ;-) Hope this helps, Frauke Zitat von TF ti...@foxmail.com: Thanks all, Merced M Leiker is right: the double IHC procedure using two primary antibodies from same species. Some expert just sent me their procedure of Heat-interval based inactivation of the primary antibody for first antigen. I am just seeking for a chemical way at room temperature. 2009-05-18 TF 发件人： Rene J Buesa 发送时间： 2009-05-18 22:31:30 收件人： tifei; gu.lang; Merced M Leiker 抄送： histonet 主题： Re: AW: [Histonet] Chemicals that inactivate the primary antibody I am completely confused. IF you have an epitope in the tissue and you react with it an specific antibody that is it. The epitope will have reacted and I do not see any way in which you can add another antibody to that initial epitope. From the reactive point of view, the epitope is blocked to react with another, unless the reaction is not totally specific and there is room for another. I just cannot imagine a way of doing this. René J. --- On Mon, 5/18/09, Merced M Leiker lei...@buffalo.edu wrote: From: Merced M Leiker lei...@buffalo.edu Subject: Re: AW: [Histonet] Chemicals that inactivate the primary antibody To: ti...@foxmail.com, gu.l...@gmx.at Cc: histonet@lists.utsouthwestern.edu Date: Monday, May 18, 2009, 9:42 AM Hi TF and all interested, I think I know what you want, but unfortunately I don't know how to answer your question (it is something I'd like answered myself!!) To re-word for the sake of all interested: You want to perform double-immunofluorescent staining using 2 primaries that were raised in the same species. An additional question for clarification is: Do you want to do this on paraffin or frozen sections? Maybe someone can help figure it out... Merced --On Sunday, May 17, 2009 2:16 AM +0800 TF ti...@foxmail.com wrote: But the heat also damage the fluorescnece... u need specific amplication kit anyway. 2009-05-17 TF 发件人： Gudrun Lang 发送时间： 2009-05-17 02:00:18 收件人： ti...@foxmail.com 抄送： histonet@lists.utsouthwestern.edu 主题： AW: [Histonet] Chemicals that inactivate the primary antibody For doublestaining the primary undergoes denaturation through a second HIER-step. The second secondary ab doesn't bind to the first primary. Therefore the binding sites must have been destroyed by heat in retrieval-buffer. Gudrun -Urspr黱gliche Nachricht- Von: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] Im Auftrag von TF Gesendet: Samstag, 16. Mai 2009 18:48 An: Histonet@lists.utsouthwestern.edu Betreff: [Histonet] Chemicals that inactivate the primary antibody Hi all, just wonder what kind of treatments/chemicals can complete block the binding of 2nd antibody to binded primary antibody on antigen? I tried HCl , but does not damage all the primaryantibody binding sites - i can still see staining pattern finally. 2009-05-17 TF ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Merced M Leiker Cardiovascular Medicine 348 Biomedical Research Building State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 lei...@buffalo.edu 716-829-6118 No trees were harmed in the sending of this email. However, many electrons were severely inconvenienced. ___ 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] Reusing specimen containers
We did it like Gary wrote: We had an extra dish washer for the containers, that produce a nice melange of formalin, damp and desinfectant. Cross contamination did not occur by washing the containers but at the point of gross section or even at the OR itself Frauke Zitat von Martin, Gary gmar...@marshallmedical.org: We cleaned bottles everyday for years and never had a problem ... having said that ... it not a good idea in the end because of the time and the exposure to the formalin. Gary -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Daniel Schneider Sent: Tuesday, April 21, 2009 11:53 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Reusing specimen containers I have no idea what CAP or JCAHO says about this but it's a very bad idea. Cross contamination happens. If you reuse containers -- and I don't care what precautions you take -- you will rarely or occasionally fail to adequately clean a container and you will end up mixing patient tissues. The cost of a specimen container is a minute fraction of the cost of processing a specimen. Ask yourself how much money you're really saving. You're scrapping for pennies. Or is this some sort of green initiative? In any case, it's asking for problems Daniel Schneider On Tue, Apr 21, 2009 at 11:43 AM, Kelly Colpitts kelly_colpi...@hotmail.com wrote: Hi Histoland! I'm just wondering what folks out there are doing about specimen containers. Is anyone cleaning them out and reusing them? Is there any CAP or Joint Commission regulations that say that all specimen containers can only be used once or can you reuse them as long as they have been thoroughly cleaned? Thanks for you all your input, Kelly _ Windows Live(tm) SkyDrive(tm): Get 25 GB of free online storage. http://windowslive.com/online/skydrive?ocid=TXT_TAGLM_WL_skydrive_042009 ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] cryoprotection possible-thanks!
Dear Histonetters, I just wanted to say a big thank you to all who responded to my cryoprotection request. We managed it to fix the samples with GA and do a sucrose step accourding to your suggestions: it worked! happy eastern to all, Frauke ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
