[Histonet] Fluorescence backgound

Monfils, Paul PMonfils <@t> Lifespan.org
Thu Aug 18 12:20:51 CDT 2005


Autofluorescence in CNS tissue, as well as some other tissues, is often due
to the presence of lipofuscin. Staining with Sudan Black B after
immunolabeling is often effective in suppressing such background
autofluorescence.

Make up 0.l% solution in 70% ethanol.  Heat to boiling, then cool and
filter.  Optimum staining time may vary somewhat from one tissue to another
and for sections of different thickness, but 10 minutes usually provides an
acceptable level of suppression, and often complete suppression of
autofluorescence.  Quickly rinse off the stain with water or buffer, several
rapid changes, and coverslip as usual.

Paul M.

> ----------
> From: 	histonet-bounces <@t> lists.utsouthwestern.edu on behalf of
> klapka
> Sent: 	Thursday, August 18, 2005 5:33 AM
> To: 	Histonet
> Subject: 	[Histonet] Fluorescence backgound
> 
> Hello all,
> 
> does anyone use a method to suppress autofluorescence of tissue due to 
> PFA fixation? I have large background in my PFA fixated nervous tissue...
> Thank you,
> Nicole
> 
> -- 
> Nicole Klapka PhD
> Labor für Molekulare Neurobiologie
> TVA, Geb. 22.22
> Heinrich-Heine Universität Düsseldorf
> Universitätsstr. 1
> 40225 Düsseldorf
> Tel: ++49 211 81 14437 
> 
> 
> 
> 
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