Re: quench autofluorescence-GFP
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From: | Scott Whittaker <sdw@biotech.ufl.edu> |
To: | Carla_Aiwohi@usgs.gov (Carla Aiwohi) |
Reply-To: | |
Date: | Mon, 30 Aug 1999 21:53:30 -0400 |
Content-Type: | text/plain; charset="us-ascii" |
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
have
>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
>much!
>
>Sincerely,
>Carla Aiwohi
>Western Fisheries Research Center
>Seattle, WA
>
>
>
<><><><><><><><><><><><><><><><><><><><><><><><><><>
Scott Whittaker ph: 352-392-1184
EM Technician fax: 352-846-0251
University of Florida email: sdw@biotech.ufl.edu
ICBR EM Core Lab web:www.biotech.ufl.edu/~emcl/
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