[Histonet] fixation and decal of cortical bone
n_hedgecock <@t> hotmail.com
Thu Aug 26 19:50:49 CDT 2004
I would like to fix, decalcify, and embed rabbit cortical bone sections in
order to do TUNEL staining for osteocyte apoptosis, but I've come across
conflicts between the literature and an actual, working protocol a colleague
has shared with me.
That being said, can you store the sample in a fixative for long periods of
time? i thought that if left in formaldehye or formalin for too long then
there would be too much cross-linking and antibodies (for IHC) could not
penetrate. Is over-cross-linking not an issue for TUNEL?
Also, don't formalin and paraformaldehyde break down into formic acid after
a period of time? I read that formic acid is no good for TUNEL, EDTA is the
decalcifier of choice.
finally, if you can't store your sample in the fixative for extended periods
of time (weeks to months), then how can I store it until I am ready to
Orthopedic Research Lab
UCD Medical Center
University of California, Davis
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