Re: nissl/nadph stain for glycerol-coverslipping
From: | "J. A. Kiernan" <jkiernan@julian.uwo.ca> |
On Fri, 13 Oct 2000, Susan Travers wrote:
> We would like to counterstain napdh-reacted brain tissue with a nissl
> stain for cell bodies. Because the nadph staining precipitates out a
> little with organic solvents, we find it best to use glycerol for
> coverslipping. Thus, we need nissl stain that contrasts with the
> purple nadph, but won't be soluble in water. Any ideas out there??
If your NADPH diaphorase method has a blue end-product (as from
nitro-BT) you need a red nissl stain for contrast. Neutral red
does a good job (try a 0.5% solution at pH 4 for about a minute;
reduce the pH or differentiate in 0.5-1.0% acetic acid if too
dark). Make your glycerol mounting medium a bit alkaline to
prevent extraction of the neutral red.
John A. Kiernan,
Department of Anatomy & Cell Biology,
The University of Western Ontario,
LONDON, Canada N6A 5C1
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