Andrea,
You might want to try using either 10% gum tragacanth made in DI water or
about a 20% solution of polyvinyl alcohol for supporting your muscle blocks.
The advantage is that both solutions are thicker than OCT and make it easier
to get the muscle oriented. Both solutions will freeze quickly. I agree that
cutting unsupported muscle, usually in cross section, is not easy, but I
find it works well if you pay attention to the basics of cryo sectioning
such as a very sharp knife, proper angles for the knife and anti-roll plate,
correct temperatures for the samples you are cutting, and a great deal of
patience. Best of luck.

John E. Tarpley 5-1-A
Associate Scientist
Amgen Inc.
One Amgen Center Drive
Thousand Oaks, CA 91320 
These Opinions are my own and not necessarily those of my employer.

-----Original Message-----
From: Andrea Grantham [mailto:algranth@u.arizona.edu]
Sent: Thursday, January 24, 2002 9:42 AM
To: histonet@pathology.swmed.edu
Subject: freezing sketetal muscle - need some suggestions


I have some labs who bring me hunks of frozen skeletal muscle for frozen 
sectioning. They clamp the muscle and snap freeze it and then store it in a 
microcentrifuge tube at minus 80 until needed. When I get the muscle I have 
to attach it to the cryostat chuck with OCT without thawing any of the
tissue.
What I have had to do is make a base of OCT and just as it is about to 
freeze over I stick only a tip of the tissue into it. However when 
sectioning I'm cutting only the tissue with no supporting medium around it 
and this ain't easy!
If any other labs are doing this and would want to share how they are going 
about it I'd be really grateful!!!
Thanks!
Andi Grantham
.....................................................................
: Andrea Grantham, HT(ASCP)     Dept. of Cell Biology & Anatomy     :
: Sr. Research Specialist       University of Arizona               :
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