"Dr. Ian Montgomery." <ian.montgomery@bio.gla.ac.uk>
<html>
Jen,<br>
<x-tab> </x-tab>Following
on from a posting several weeks ago, attach the muscle to a <b>cork disc
</b> with OTC before freezing. This will save you using a cryostat chuck
which in a multi-user facility might be in short supply. Plus, why store
a valuable chuck when a wee cork disc can be attached to the chuck many
times without the risk of muscle thaw. <br>
Ian.<br>
<br>
<br>
<blockquote type=cite class=cite cite>Date: Thu, 25 Jan 2001 10:59:02
+0000<br>
From: "t.hacker@har.mrc.ac.uk"
<T.Hacker@har.mrc.ac.uk><br>
Subject: Re: cryo-preservation<br>
To: "Philopena, Jennifer"
<jennifer.philopena@canji.com><br>
Cc: "'Histonet'" <Histonet@pathology.swmed.edu><br>
<br>
Date sent:
<x-tab> </x-tab>Wed, 24
Jan 2001 17:23:59 -0500<br>
From:
<x-tab> </x-tab>"Philopena,
Jennifer" <jennifer.philopena@canji.com><br>
Subject:
<x-tab> </x-tab>cryo-preservation<br>
To:
<x-tab> </x-tab>"'Histonet'"
<histonet@pathology.swmed.edu><br>
<br>
> Happy Chinese New Year to all in Histoland.<br>
> <br>
> I am doing fresh frozen sections of muscle and am getting freezing
artifact.<br>
<br>
Jen, I would avoid any form of "cryo-protection" with muscle as
this <br>
tends to provide the tissue with enough time to develop ice crystal
<br>
artefact before freezing (producing those nasty little holes within
<br>
the fibre). Expose the fibres as best you can (small block) and <br>
immerse in pre cooled (LN2) isopentane, preferably on the cryostat <br>
chuck (specimen holder). If you attach the tissue to the chuck after
<br>
freezing avoid any thawing and re-freezing as this will introduce <br>
even worse artefact.<br>
The above observations are the result of personal experience and a <br>
number of frustrating hours attempting to get perfectly preserved <br>
muscle fibres.<br>
Best of luck/skill,<br>
Terry.<br>
Terry Hacker,<br>
Medical Research Council,<br>
Harwell,<br>
Didcot,<br>
Oxfordshire, OX11 ORD<br>
01235 834393 x360</blockquote>
<x-sigsep><p></x-sigsep>
<font color="#0000FF">Dr. Ian Montgomery,<br>
West Medical Building,<br>
University of Glasgow,<br>
Glasgow,<br>
G12 8QQ.<br>
Tel: 0141 339 8855. Extn:6602.<br>
Fax: 0141 330 2923<br>
e-mail: ian.montgomery@bio.gla.ac.uk</font></html>