RE: Spermatozoa stains (New and other fuchsines)

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From:"Nader, Alexander" <>
To:"'J. A. Kiernan'" <>

I remember when I was a student we used a nigrosin(e)-eosin method. maybe
nigrosin(e) is better known as acid black 2.
I found some information about this stain in

But I thaught that nigrosin(e) is not a basic, but a acidic dye. 

> -----Original Message-----
> From: J. A. Kiernan []
> Sent: Saturday, August 05, 2000 7:25 AM
> To: Roberta Horner
> Cc: Histonet
> Subject: Re: Spermatozoa stains (New and other fuchsines)
> On Thu, 3 Aug 2000, Roberta Horner wrote:
> > Does anyone have a procedure for a stain for spermatozoa?  
> Or could someone
> > tell me if new fuchsin is also known by another name? I  
> have Berg's method
> > but it uses new fuchsin.  I need this as soon as possible 
> for a ram sale.
>   New fuchsine is one of the dyes that compose basic fuchsine. The
>   others are pararosaniline, rosaniline and magenta II. Of these,
>   only pararosaniline and new fuchsine are commercially available
>   as single "pure" dyes. 
>   For most purposes it doesn't matter if you use one of the pure 
>   dyes or a mixture. For the aldehyde-fuchsine stain you must use 
>   pararosaniline or a basic fuchsine that is mostly pararosaniline
>   (Mowry & Emmel 1977 J Histochem Cytochem 25:239). The only other
>   methods I know about for which a single dye is advisable (possibly
>   necessary) are those for which you make a diazonium salt for use 
>   in esterase or peptidase enzyme activity histochemistry (hexazonium
>   pararosaniline or hexa-azotized new fuchsine).
>   Both basic fuchsine and pararosaniline are available as certified
>   dyes. New fuchsine (CI 42520) is not certified as such. A dye
>   certified as basic fuchsine could contain only pararosaniline or
>   new fuchsine, or it could be a mixture of 2, 3 or all 4 dyes (often
>   with a high percentage of rosaniline). Rosaniline (C.I. 42510) and
>   magenta II (has no C.I. number) can be made only by separation
>   from mixtures, which isn't commercially worthwhile. They are not
>   sold as single dyes because there would be no known use for them.
>   In this email I have used -ine rather than -in endings for the
>   names of the dyes because this is the chemically correct spelling
>   in English, and is used worldwide in the literature of dyes.
>   Most catalogues still use fuchsin, pararosanilin etc., following
>   an older (50 yrs +) tradition in which these dyes were derivatives 
>   of the German "Anilin" rather than the English/French/US "aniline."
>   Amines have ended in -ine for a century or more in the English
>   tradition.
>   The -in ending is used for substances that are _not_ derived from
>   organic bases (amines).  Eosin, erythrosin and haematoxylin are
>   good examples.  So are dextrin and insulin.  
>   Sorry if this doesn't tell you how to stain spermatozoa. Someone
>   else is sure to do that. This should put you & others in the
>   picture about the various basic fuchsine dyes. 
>  John A. Kiernan,
>  Department of Anatomy & Cell Biology,
>  The University of Western Ontario,
>  LONDON,  Canada  N6A 5C1

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