Re: nissl/nadph stain for glycerol-coverslipping

From:"J. A. Kiernan" <>

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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