Re: [Histonet] GFP in Paraffin Sections?

From:Gayle Callis

Paul,

You may have success or you may not.  We cannot maintain our eGFP with 
paraffin processing, or with any frozen section fixed with 
solvents.   Alcohols may ruin the GFP and its fluorescent properties, 
however there are those who have had success.  One can always come back 
with antiGFP (Rockland makes are very nice goat antiGFP) and then detect 
the goat antiGFP with a Donkey antiGoat-FITC.

It is certainly worth a try to do what you suggest and try both ways or 
mounting medias.  If your tissues are FFPE, then you will be battling green 
autofluorescence too.  I have a review of autofluorescence with GFP in mind 
I am sending to you via private email, a very nice publication.

At 11:25 AM 10/24/2005, you wrote:
>Does anyone know if GFP (Green Fluorescent Protein) maintains its
>fluorescence through paraffin processing?  Can the fluorescence be viewed in
>an otherwise unstained paraffin section?  After deparaffinization, should
>the section be coverslipped with an aqueous medium, or the standard medium
>used for paraffin sections?
>
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Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)



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