unfixed cryo sections left at room temp...

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From:"L. Gibbs" <lgibbs@u.washington.edu>
To:Histonet <HistoNet@pathology.swmed.edu>
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Hi Histonetters-

When I do cryosectioning on unfixed specimens, I usually let
sections air-dry for a couple minutes and then store at -80C
until needed. For most applications we would then remove slides
from freezer and allow condensation to evaporate before doing a
post-fix. On occasion, I have done ICC on slides without
fixation.

More often, unfixed cryosections are cut then post-fixed within a
few minutes and air-dried. These fixed slides may be stored at
room temperature for a day or at 4C or back to the freezer for
long-term storage. 

The question:

I just received some un-fixed cryo slides that have been at
r.t. for 24 hours.
They are for cytokeratin (various antibodies) ICC.

How long can unfixed cyro-sections be left at r.t. before the
tissue and epitope integrity are jeopardized?

Thanks much!

Lorraine Gibbs




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