RE: Mast cells/Bismarck Brown

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From:"Gary W. Gill" <garywgill@email.msn.com>
To:"Tony Henwood" <henwood@mail.one.net.au>, "HistoNet Server" <HistoNet@pathology.swmed.edu>, "A G DU TOIT" <AGDT@GERGA.SUN.AC.ZA>
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Date:Thu, 16 Sep 1999 21:14:52 -0500
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Make sure the pH of the staining solution is alkaline.  Bismarck brown Y is
a basic dye, which means not that it is alkaline, but that it carries a
cationic dye that carries a positive charge.  By ensuring an alkaline pH,
even slightly, there are more negative charges on the amino acids that
constitute the constituent proteins, thus promoting uptake of basic dyes.  I
have no experience staining mast cells, so my suggestion may be groundless.
It is nonetheless fundamentally sound.

Gary W. Gill

-----Original Message-----
From: Tony Henwood [mailto:henwood@mail.one.net.au]
Sent: September 15, 1999 6:16 PM
To: HistoNet Server; A G DU TOIT
Subject: Re: Mast cells/Bismarck Brown


Dear Andre,

> I am looking for a stain specifically for Mast cells.  We have been
> doing the Bismarck Brown for some time, but unfortunately no
> literature available.  Surely somebody might just be able to help.
> Thanks a million in advance, I am looking forward to hear from all of
> you.
>
Most Mast cells stain metachromatically, so a toluidine blue (0.1%
aqueous) will do the trick. Allow stained sections to air dry prior
to coverllipping (ie no alcohol). Churukian has a marvelous technique
that was published some time ago in J Histotechnology.

Aqueous alcian blue (1% in 0.5% HCl) (pH 1) will also stain them.
This is mainly due to their being rich in heparin sulphate. Try one
of the Sigma dyes - they tend to  dissolve better at this pH.

My histonet summaries contain a page on Mast cell controls which may
be of use.
Regards, Tony
.
Tony Henwood
Senior Scientist
Anatomical Pathology
Royal Prince Alfred Hospital
Sydney, AUSTRALIA

http://www2.one.net.au/~henwood
http://www.pathsearch.com/homepages/TonyHenwood/default.html





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