Date sent:      	Wed, 24 Jan 2001 17:23:59 -0500
From:           	"Philopena, Jennifer" <jennifer.philopena@canji.com>
Subject:        	cryo-preservation
To:             	"'Histonet'" <histonet@pathology.swmed.edu>

> Happy Chinese New Year to all in Histoland.
> 
> I am doing fresh frozen sections of muscle and am getting freezing artifact.

Jen, I would avoid any form of "cryo-protection" with muscle as this 
tends to provide the tissue with enough time to develop ice crystal 
artefact before freezing (producing those nasty little holes within 
the fibre). Expose the fibres as best you can (small block) and 
immerse in pre cooled (LN2) isopentane, preferably on the cryostat 
chuck (specimen holder). If you attach the tissue to the chuck after 
freezing avoid any thawing and re-freezing as this will introduce 
even worse artefact.
The above observations are the result of personal experience and a 
number of frustrating hours attempting to get perfectly preserved 
muscle fibres.
Best of luck/skill,
Terry.
Terry Hacker,
Medical Research Council,
Harwell,
Didcot,
Oxfordshire, OX11 ORD
01235 834393 x360