Your "new doctor" is absolutely wrong. You do not need to formalin fix or sucrose wash before freezing the specimen in OCT. You can go directly from the fresh specimen → OCT freezing → cutting. You are partially right about the "cross-linking" of the antigens; only "partially" because cross-linking requires about 20 hours and you will never wait so long to make the FS. In any event, it is not necessary to do what your "new doctor" says; it is evident that s/he is really "new". René J.
--- On Thu, 6/23/11, Joel Reichensperger <jreichensper...@siumed.edu> wrote: From: Joel Reichensperger <jreichensper...@siumed.edu> Subject: [Histonet] Frozen tissue question To: histonet@lists.utsouthwestern.edu Date: Thursday, June 23, 2011, 10:12 AM We have a new doctor in our lab who swears that all frozen tissue must be fixed in formalin with a subsequent sucrose treatment before freezing in OCT because not fixing it will cause the structures to be distorted and you can't get good antibody attachment. In my previous experience, we have done this with tissue that came from an animal that was perfused with formalin before the tissue was removed. However, the majority of my previous frozen specimens were flash frozen in OCT and fixed after sectioning. It is also my understanding that fixation in formalin can cause poor antibody detection because of cross-linking caused by the formalin. I would like to hear some other opinions on this please. The particular specimen we will be working with is skin. Thanks in advance, Joel Reichensperger -- Joel Reichensperger Researcher II Southern Illinois University Plastic Surgery Institute jreichensper...@siumed.edu 217-545-7309 (Office) 217-545-1824 (Fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
