[Histonet] Frozen Rat Heart

[Sebree Linda A]

Sharon,

I used to freeze rabbit hearts (a LITTLE bigger than rat) this way.  

Pour isopentane into a metal beaker inside a styrofoam container filled
with liquid nitrogen to part way up the sides of the beaker.  When the
isopentane starts to freeze on the bottom of the beaker it is ready.
Place cross sections of fresh tissue (or the whole rat heart) on a round
of cork with gum tragacanth, orienting it to the way you may be
sectioning it if that's its destiny.  With a LONG forceps or hemostat,
lower cork round, heart side up, into the isopentane.  It's important to
lower the tissue slowly to prevent cracking but not so slow as to cause
freeze artifact.  Complete freezing will only take a couple minutes if
that.  To section tissue, if that's the next step, freeze the bottom of
the cork round onto a chuck using OCT.  In this way, you insulate the
tissue from the RT or cold OCT preventing any chance of thawing of your
tissue.

Hope this helps you in your situation,
  
Linda A. Sebree
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
DB1-223 VAH
600 Highland Ave.
Madison, WI 53792
(608)265-6596


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Shaw,
Sharon
Sent: Thursday, April 22, 2010 10:06 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Frozen Rat Heart

Does anybody have a protocol on freezing rat heart that you can share
with me.
Thanks,
Sharon
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