[Histonet] Snap Freezing Tissue

[Adam .]

I learned to freeze tissue by taking a beaker and filling it with
2-methylbutane and then placing that beaker in a liquid nitrogen bath. You
then take place your cryomold, hold it using some long forceps, and place it
on the top of the 2-methylbutane. It will freeze the entire thing in about
10 seconds. Once it's frozen, you dip the entire mold into the
2-methylbutane. Seems to work well for me.

Adam

On Wed, Feb 24, 2010 at 4:13 PM, Andrea T. Hooper <
andreahooper <@t> rocketmail.com> wrote:

> 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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