----- Original Message -----
From: A G DU TOIT <AGDT@GERGA.SUN.AC.ZA>
To: HistoNet Server <HistoNet@pathology.swmed.edu>
Sent: September 13, 1999 5:36 AM
Subject: Mast cells/Bismarck Brown


> Dear All,
> 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.
> Andre du Toit
> Dept Anatomical Pathology
> Tygerberg Hospital
> Cape Town
> South Africa.
>
>


I use aldehyde-toluidine blue.  Make it the same as aldehyde fuchsin, just
use toluidine blue instead of basic fuchsin.  Ripen it for about a week at
room temperature.  It is stable for a very long time if used for mast cells.
I have noted that the sample of toluidine blue does make a difference.  The
best seems to be from Fisher Scientific.  You may need to try several
others.  When you find one, keep lots.

1. Sections to absolute ethanol.
2. Place in aldehyde TB for 1-2 hours.
3. Wash with abs. ethanol.
4. Wash with water.
5. Counterstain nuclei with nuclear fast red 5 mins
6. Contrast stain with 0.1% tartrazine in water for 15 seconds.
7. Dehydrate, clear and mount.

Mast cell granules - deep blue
Nuclei - red
Background - pink and yellow


*Nuclear fast red*
Nuclear fast red 0.5 g
Pot. or amm. alum 25 g
Water 500 ml
Mix, dissolve, boil 2 minutes, cool, filter.