RE: GFP in tissue sections

From:"Montague, Donna C"

EGFP is stable in paraffin processing. Prepared slides need only be deparaffinized with three changes of xylene 5 minutes each, covered with Vectashield (Vector laboratories) or Prolong Antifade (Molecular Probes) mounting media and coverslipped with a No 1 thickness glass. The problem with fluorescence visualization of eye structure maybe autofluorescence from retinal pigments. Choose your excitation wavelength wisely or visualize using anti-GFP antibody detection immunohistochemically. Hope this helps,
Donna Montague, M.S., Research Associate
University of Arkansas for Medical Sciences
Center for Orthopaedic Research
Departments of Physiology & Orthopaedic Surgery
4301 W. Markham St. # 505
Little Rock, AR 72205
(501) 603-1239 

-----Original Message-----
From: Pillai, Mahesh (NIH/NEI) [mailto:PillaiM@NEI.NIH.GOV] 
Sent: Monday, December 16, 2002 10:32 AM
To: 'MontagueDonnaC@uams.edu'
Subject: GFP in tissue sections
Importance: High


Hi Dr. Montague,
I am attempting to look at the ocular tissue expression of EGFP in mouse models for my studies. I was wondering whether you can give me some input on whether I should use frozen sections or paraffin embedded sections after fixation for examining under fluorescent microscope.

Thanks and regards,

Mahesh

Dr S Mahesh MD,FRCS
Laboratory of Immunology 
National Eye Institute
Bldg 10 Room 10N113
10 Center Drive
Bethesda, MD 20892
phone: 301-496-1322
fax: 301-480-1122





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