[Histonet] bleaching problem

From:"tiang yeepeng"

Dear all,
I  would like to ask if anyone ever experienced fast bleaching of IF staining of your samples after the exposure to light emitted from the fluorescence microscope. I am staining with a FITC conjugated secondary Ab (Jackson Immunolab) and mounted with ProLong Gold (Molecular Probes). The fluorescence got very dim after 20-30sec of exposure. Though I got back the signal after removing it from the fluorescence microscope awhile but the signal was very much weaker from the very first time I saw in the microscope. I am unable to find out what is the cause. Can anyone please advise? 
Thanks alot!

With regards,
TIANG YEE PENG
Department of Medical Microbiology,
Faculty of Medicine,
University of Malaya, Malaysia
 
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