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 brought up 282 hits!
There's a good account by Roy Ellis at
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1

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