[Histonet] post-fixing quick frozen tissues for IHC

[Patsy Ruegg]

I have the same issues, I get tissue quick frozen without any OCT or
surcrose cryoprotectant from the molecular biologists all the time,
sometimes the tissue is not even quick frozen it is just taken fresh and
stuck in a tube in the -80dc freezer and the morphology is terrible.   There
was a technique a year ago in Histologic about thawing and refreezing poorly
prepared samples, I have tried this with some improvement but it is never as
good as doing it right in the first place.  I would be interested in others
approach to this issue as I don't see an end to getting samples not properly
prepared for frozen sectioning.
Patsy 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of phil tsai
Sent: Tuesday, June 28, 2005 5:26 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] post-fixing quick frozen tissues for IHC

Hello all,

   I was wondering if anyone has had any problems with tissues that were
quick frozen before being being fixed and cryoprotected?  We usually obtain
our tissues fresh, fix in some sort of paraformaldehyde solution (Zamboni's,
Lanas), cryoprotect in sucrose solution for around 1 week, and then quick
freeze.  However, for our positive controls, we are only able to obtain
quick frozen tissues that haven't yet been fixed.  I suppose that this might
affect the achitecture a bit, but does anyone have any knowledge of whether
immunohistochemistry might be affected?  We are looking for nerve
innervation, and will be obtaining muscle and skin for these purposes....

-Phil



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