Re: quenching neuronal autofluorescent

From:Cathy Gorrie

Re: quenching neuronal autofluorescent
Dear Carol,

I haven't tried this yet but it was sent to me by one of my colleagues who has. She didn't send the paper but I could get hold of it. Worth a try.

Attached is a paper describing the reduction of autoflourescence using Sudan Black B. The protocol I used is for immunostaining.  After the incubation with secondary antibody, slides are washed 3 times, than are treated with Sudan Black B, 0.5% in 70% Ethanol (Sigma, catalog # S0395) for 8 minutes, washed quickly with PBS+0.05% Tween-20, mounted with anti-fade and covered with coverslip.


At 2:40 PM -0600 12/2/03, Carol Bobrowitz wrote:
We are using paraformaldehyde to perfuse goat brainstem for indirect
fluorescent (FITC) immunohistochemical analysis.
The frozen sections (25um) are mounted on chrome-gelatin treated slides. 
After staining a slide thru the immuno procedure with absolutely no antibody
(primary or secondary) application, we found high levels of neuron-specific
Is there some chemical we can apply or treatment we can perform to quench
the autofluorescence?
Any help or suggestions will be greatly appreciated.

Carol Ann Bobrowitz

Department of Physiology
Medical College of Wisconsin
Milwaukee, Wisconsin
(414) 456-8179
FAX (414) 456-6546

Catherine Gorrie
School of Medical Sciences,
University of New South Wales
Sydney, N.S.W. 2052

Phone: 61-2-9385 2462
Fax  : 61-2-9313 6252

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