Re: quench autofluorescence-GFP

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From:Scott Whittaker <> (Carla Aiwohi)
Date:Mon, 30 Aug 1999 21:53:30 -0400
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I will assume you have not fixed the tissue in glut. You might try tol
blue. We have even doubled up and used 0.1% tol blue in borohydride.
Another option I have heard of but  not tried is using Evans Blue to shift
the autofluor. to emit red while your signal stays yellow/green. I do not
know how or what it is made in nor do I know how it works and I have never
found a reference. If anyone else has used this I would appreciate hearing
back as I have been having a murderous time labeling some marine fungi
which light up under everything I have thrown at them. Good luck

At 02:45 PM 8/30/1999 -0600, you wrote:
>Hi everyone:
>I am desperately seeking any suggestions as to quenching autoflourescence in
>tissues that have been 'treated' with green fluorescent protein (GFP)?  We
>6-8 micron frozen sections in which the GFP (if present) cannont be
>distinguished from the background autofluorescence.  Clontech suggests sodium
>borohydride in PBS.  As the slides will be looked at tomorrow or Wed., I am
>looking for a substitute/alternative to the sodium borohydride which I don't
>have.    Any and all suggestions will be greatly appreciated.  Thank you very
>Carla Aiwohi
>Western Fisheries Research Center
>Seattle, WA 

Scott Whittaker                   ph: 352-392-1184
EM Technician                    fax: 352-846-0251
University of Florida   email:
ICBR EM Core Lab

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