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Dear Hege,

GFP could be quenched by aldehyde fixation, on the other hand you need to
fix the tissue to avoid wash out of GFP as it is mostly located in cytoplasm
(well, depending how its expression is controlled). Poor fixation may result
in a loss of the GFP from the tissue even if you think of not using
permeabilization etc. I would recommend fixing your tissue before
cryosectioning and then use anti-GFP antibody to enhance native GFP (if you
find the signal too weak). I remember I found an extensive discussion on
this issue here in the list or elsewhere in microscopy forums.

good luck

Kirill Ukhanov
University of Florida

On Fri, 7 Mar 2008 16:22:06 +0100, Hege Avsnes Dale
<[log in to unmask]> wrote:

>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Hi.
>Do anyone have a protocol for good preservation of GFP in tissue after
>freeze sectioning?
>The tissue is a tumor with EGFP-expressing cells and need only a
>fixation that will preserve GFP (no permabilization  needed). Would it
>also be a better idea to fix before sectioning?
>
>Hege
>=========================================================================