Re: freezing fixed tissues

From:"J. A. Kiernan"

> Garry Ashton wrote:
> I am about to freeze down some tissues that have been fixed in
> 10%NBF.
> Is taking the tissue through a sucrose gradient and freezing
> slowing in OCT the best way to achieve reasonably good
> morphology, only I remember doing this several years ago and
> the morphological preservation was quite poor.

Freezing slowly is always bad.
Soaking in 30% sucrose will reduce the
sizes of the ice crystal holes; they
should be small enough not to be
visible by light microscopy.
There's lots of stuff on the web about
this. Looking for "sucrose" on 
www.histosearch.com brought up 282 hits!
There's a good account by Roy Ellis at
http://home.primus.com.au/royellis/fr.htm
-- 
-------------------------
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
   kiernan@uwo.ca
   http://publish.uwo.ca/~jkiernan/



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