[Histonet] Snap Freezing Tissue

[Andrea T. Hooper]

I have used the "Gayle Callis" method of freezing tissue for histological applications for years with great success :) The tissues look fantastic. 
 
Essentially float out a metal pan (like the ones you sterilize surgical tools in) on a liquid nitrogen bath (in a styrofoam container or large dewar). Place fresh or cryoprotected tissue into cryomold (I like the Tissue Tek ones mainly) and add sufficient OCT. Place mold onto metal tray and close lid of dewar or styrofoam container. Let freeze. When frozen wrap in foil and place into bitran bag, place into -80 deg C for long-term storage.
 
If snap freezing tissue for RNA/protein extraction I use the classic isopentane cooling method.
 
Andrea T. Hooper


--- On Wed, 2/24/10, Laurie Colbert <laurie.colbert <@t> huntingtonhospital.com> wrote:


From: Laurie Colbert <laurie.colbert <@t> huntingtonhospital.com>
Subject: [Histonet] Snap Freezing Tissue
To: histonet <@t> lists.utsouthwestern.edu
Cc: "Steve Pike" <stevep <@t> mikronet.com>
Date: Wednesday, February 24, 2010, 9:54 PM


I was wondering how others are snap freezing tissue?  We have been using
a histobath (a refrigerator device that cools isopentane for freezing
tissue), but it has died.  I would love to have another Histobath, but
they are no longer carried by the vendor that I originally bought mine
from.  Does anyone know where I can get either a new or refurbished
Histobath (vendor calls are welcome)?  I'm also interested in other
freezing techniques.



Laurie Colbert

Huntington Hospital

Pasadena, CA

(626) 397-8620 

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