Re: quenching neuronal autofluorescent
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.
Cathy
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
autofluorescence.
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
cbobrowi@mcw.edu
--
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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
e-mail: c.gorrie@unsw.edu.au
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