Patti, we do the Dahl's calcium stain very often! (AFIP, 3rd edition) The
procedure follows. The only thing to be careful of is the pH of the
solution. The Alizarin red solution will get very thick with time, but as
long as the pH is correct, it works well for us:

Fix:           10% NBF or 95% Ethyl alcohol
Technique:      5 microns

1% Alizarin Red S Solution:
          Alizarin Red S...... 1.0g
          Distilled water...... 100.0 ml
          Mix well, add 10 ml of 0.1% ammonium hydroxide, slowly with
constant stirring. pH to 6.4 using more ammonium        hydroxide if
necessary. The book says that the solution is stable for 1 month. We find
that if you maintain the 6.4       pH, it lasts almost indefinitely.

Procedure:     Deparaffinize to 95% alcohol
          Alizarin Red S for 5-10 minutes
          Distilled water rinse, 6 dips  (I have used tap water without
negative results)
          1% light green counterstain, 1 minute
          Distilled water rinse, 6 dips
          Dehydrate to xylene
          Synthetic mount

Results:  Calcium salts - intense reddish-orange

This is a very reliable and easy stain!





"Bourassa, Patricia" <patricia_bourassa@groton.pfizer.com> on 10/06/98
12:17:49 PM

To:   "Histonet @Pathology.Swmed.Edu (E-mail)"
      <histonet@pathology.swmed.edu>
cc:    (bcc: Lori Miller/WLGORE)
Subject:  Alizarin Red




Hi, There!

Has anyone had any experience using Alizarin Red to stain for Calcium
deposits?  I was wondering if there were any fixation and/or embedding
restraints.  Also, does anyone have any good protocols for a specific
staining method for calcium?

Thanks!!

-Patti Bourassa
 Pfizer, Groton