Re: [Histonet] GFP

From:Gayle Callis

This has been discussed on Histonet, go to Archives and search, look for
Donna Montague, she has great success with paraffin work unless you are
doing frozen sections.   

Formalin and 4% paraformaldehyde will work as a fixatives.  You need to
know the difference between autofluorescence and GFP, hopefully the GFP
will fluoresce at maximum level to overcome the autofluorescence.  So you
need to have a control, naive or normal (contains no GFP) rat duodenum,
fixed at same time as your experimental animal, and look at this tissue so
you know how autoflourescence appears.    

Do not use an H&E, GFP is fluorescent and you need to examine without any
staining to see GFP, hopefully not a wild type GFP but eGFP, or enhanced GFP. 

At 08:33 PM 11/10/2003 EST, you wrote:
> I need to use this procedure to visualize a vector that will be injected
>into rat duodenum. I would appreciate any hints on fixation and staining
>procedures procedure that anyone may want to share.
> lin
Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)

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