[Histonet] flash frozen tissue

[Ingles Claire]

We are a Mohs clinic and I had done a study on the feasability of IHC on frozen skin. I cut slides as normal then fixed in acetone before using Biocare's Mach 3 kit (Alk. phos. with Red chromogen) with predilute antibodies. They mostly turned out well, except for the S100. I don't believe that one turns out satisfactory in frozen tissue no matter what you do. I did a MART-1 stain, HMB-45 (predilute from Innovex), and Tyrosinase. There was very little background. Biocare will be more than happy to help with any questions you might have.  
Claire  

________________________________

From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Kalleberg, Kristopher
Sent: Wed 7/29/2009 9:31 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] flash frozen tissue



Hello All,

I will be running a study upcoming and we have decided not to run FFPE
as we normally do and have decided to use flash frozen.  My big concern
is what is the proper way to fix all of these SKIN samples to use for
IHC.  Any recommendations as to if prefix the samples in sucrose and
then flash freeze in OCT is best or any recommendations as to if post
fixing (in acetone??) the already sectioned, flash frozen sample in OCT
is best.  Any and and all help will be greatly appreciated.  Thank you.

Kris Kalleberg   
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