[Histonet] Davidson's fixative and IHC question
jaylundgren at gmail.com
Mon Feb 22 13:53:09 CST 2016
Today I learned what Davidson's fixative is! Thanks!
Anyway, so it's a strong alcoholic formalin. The answer to your question
is; yes. You need to be careful of overfixation, thus masking the epitope
" Of the many pre-analytical variables which affect IHC and ISH
results, fixation is probably the most significant, impacting many other
variables such as antigen retrieval and epitope binding. Unfortunately, to
date, no single fixative has proven to be ideal for all targets and
detection methods. However, it is generally more deleterious for tissue to
be ‘underfixed’, rather than ‘overfixed’."
Make sure not to leave your specimen in Davidson's for more than 24 hrs,
and then in formalin for *less* than 72 hrs and you should be OK. If
staining is unsatisfactory you might have to tinker with your antigen
retrieval protocol (longer) or primary Ab incubation time (longer).
Jay A. Lundgren, M.S., HTL (ASCP)
On Mon, Feb 22, 2016 at 11:45 AM, Judi Ford via Histonet <
histonet at lists.utsouthwestern.edu> wrote:
> Hi Everyone. Hope you all had a really good weekend. Thanks for the
> replies to my double ihc question.
> I do have another question; this one is from a friend of mine. Her client
> wants to do ihc on rabbit eye tissue. The tissue will be fixed in
> Davidson's fixative for 24 hours then in 10% NBF. Will this have affect
> future ihc projects?
> Histonet mailing list
> Histonet at lists.utsouthwestern.edu
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