Re: Tissue in OCT - how to "Thaw??? HELP!!

From:John Kiernan (by way of Histonet)

Thawing will not cause damage. Freezing might have
damaged the cells if it was not done
properly. The harm results from ice crystals that
leave holes in the tissue when the ice melts. If
the crystals form slowly they make holes big enough
to see with a microscope. In really badly frozen
tissue the holes are about the same size as the
cells and are so numerous that the architecture of
the tissue is not recognizable in sections. There
is nothing you can do to reverse damage already
caused by ice crystals.

If the ice crystals are too small to see, then the
tissue will be fine for light microscopy, and it
will not matter whether you thaw quickly or slowly.
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
Bruce Abaloz wrote:
> Hello Histonetters/Colleagues,
> I have a piece of 'Pouch Young' (Wallaby) tissue approx. 1mm. in size
>that was fixed in 4%  PFA then frozen in OCT. I now want to do 'WHOLE
>MOUNT IMMUNO. & am wondering what procedure IS best to 'thaw' the tissue
>with MINIMAL damage to the cells - OR anyone with previous experience RE:
>this situation can offer advice. Thankyou in advance,
> Gratiana ( )
> --
> BRUCE ABALOZ                           PH:61383446282
> HISTOLOGIST                            FAX:61383447909
> DEPT.of ZOOLOGY                        EMAIL:
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