[Histonet] Snap Freezing of Lymph Nodes for RNA testing

Thu Apr 17 07:34:31 CDT 2008

For snap freezing muscle biopsies to send on dry ice we use a 100% Alcohol
or Acetone mixed with dry ice slurry.
1. Wrap dry ice pellets in a towel, pulverize the dry ice with a hammer than
pour the powder dry ice into a 200 ml beaker.
2. Slowly add 100% alcohol and stir the mixture. At least 80% of the total
volume of the slurry should consist of dry ice and only 20% alcohol. (
Acetone can replace 100% alcohol in the slurry)
3.When the slurry stops bubbling, the temperature has fallen to around -70C,
it is then suitable for flash freezing. Keep adding dry ice as needed to
reach this point. The final product should resemble a "snowcone".
4. Hold the biopsy specimen with forceps, and plunge tissue into the slurry
quickly. Swirl the specimen in the slurry for 10-15 seconds, remove and
quickly blot dry with an absorbent towel to remove excess alcohol.
Immediately place tissue in a pre frozen specimen container that has
prechilled on dry ice. A well frozen specimen should have a white chalky
color.
5. Place cover on container. (puncture lid prior to allow excess alcohol to
evaporate.)
6. Ship away on dry ice or store in a freezer that goes down to -70 C. We
have a Jewitt freezer.

Hope this helps you. It is a inexpensive alternative. Credit goes to Mayo
Medical Services where this system is recommended for shipping to them.

----- Original Message ----- 
From: "Norm Burnham" <Norm.Burnham <@t> propathlab.com>
To: <gayle.callis <@t> bresnan.net>; <histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, April 16, 2008 9:31 PM
Subject: Re: [Histonet] Snap Freezing of Lymph Nodes for RNA testing

> Hi Gayle,
> Thank you for your advice. We want to avoid LN and Isopentane if at all
> possible.  We are looking for a simple, inexpensive, easy to use, and
RNAse
> free method to preserve, stabilize and transport tissue for RNA studies.
> Does anyone have an alternative to LN or Isopentane bath for snap freezing
> followed by shipment on dry ice, or an alternate way, to stabilize tissue
for
> periods ranging from a few hours to perhaps as long as 72 hours?
> Thanks and best regards,
> Norm Burnham
>
> ----- Original Message -----
> From: Gayle Callis <gayle.callis <@t> bresnan.net>
> To: Norm Burnham; histonet <@t> lists.utsouthwestern.edu
> <histonet <@t> lists.utsouthwestern.edu>
> Sent: Wed Apr 16 19:20:36 2008
> Subject: Re: [Histonet] Snap Freezing of Lymph Nodes for RNA testing
>
> It may be better to use a transportable Dewar flask for just the liquid
> nitrogen.  I don't think a doctors office will want to handle isopentane,
> too volatile, flammable and explosive.  Contact or maybe those on Histonet
> in dermatopathology offices have ways to do this for biopsies they
collect,
> freeze and ship.
>
> Good luck
>
> Gayle M. Callis
> HTL/HT/MT(ASCP)
> Bozeman MT 59715
>
>
> ----- Original Message ----- 
> From: "Norm Burnham" <Norm.Burnham <@t> propathlab.com>
> To: <histonet <@t> lists.utsouthwestern.edu>
> Sent: Wednesday, April 16, 2008 12:50 PM
> Subject: [Histonet] Snap Freezing of Lymph Nodes for RNA testing
>
>
> Hi Histonet,
>
>
>
> Does anyone have a simple, inexpensive and very portable method for snap
> freezing lymph nodes in preparation for shipment on dry ice?  We have
> considered Liquid Nitrogen and Isopentane bath, but I am looking for a
fully
> portable method that can be handled in a doctors office by their staff.
>
>
>
> Thanks,
>
>
>
> Norm Burnham
>
>
>
> ____________________________________
>
> Norm Burnham
> Director of Laboratory Operations
> ProPath - The Leader in Pathology Services(r)
>
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> Dallas, TX  75247
> 214.237.1602 Office
> 214.237.1802 Fax
>
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>
> Email:  norm.burnham <@t> propath.com <mailto:norm.burnham <@t> propath.com>
>
>
> To learn more about ProPath, please visit www.ProPath.com
> <http://www.ProPath.com>
>
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