Re: Immunohistochemistry on brain sections

From:Carrie Kyle <ckyle@cvdls.ucdavis.edu>

hi marilyn,
i've a couple of suggestions for you.  if you are doing formic acid treatment BEFORE processing, be sure to neutralize in running water for at least 2hr. prior to loading the tissues on your processor.  also, we neutralize the slide sections after the 5min. formic acid treatment in Tris buffer pH 7.5....3 changes of 5min each.  we rarely have difficulty with neuro tissue coming off when doing scrapie (unofficial, of course, "real" testing can only be done in the US by "official" USDA designated labs) or CWD testing.
i'm aware of a gentleman by the name of Valachandra (probably mangled the spelling) who is doing surveillance testing in Canada.....he might be helpful to you too.  i can get contact info for you if would like.
good luck,
carrie kyle-byrne
ucdavis-cahfs


>>> <marilyn.johnson@gov.ab.ca> 03/06/01 03:09PM >>>
Hi Histonetters,
I am having difficulty with brain sections adhering to the slides during
Immunohistochemistry, even using the Aminosilane-coated slides, dried
overnight at 60 degrees. These sections are hydrated, placed in a 96% formic
acid bath for 5 mins. followed by autoclaving for 25 mins. The slides are
then stained with the Dako Autostainer.  Any suggestions would be greatly
appreciated.
Thanks in advance.

Marilyn Johnson
Agri-Foods Surveillance Systems
Alberta Agriculture
Edmonton, AB.







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