Re: [Histonet] Eosinophil Staining

From:Bryan Llewellyn

The sirius red method you describe was originally published as a technique 
for amyloid.  It was only incidentally noted that it demonstrates 
eosinophils well.  You can leave out the sodium chloride if it is used 
solely for that purpose.  It is found at:, and the dye 
is identified specifically at:

Bryan Llewellyn

----- Original Message ----- 
From: "Griffin-Reyes, Michelle A" 
Sent: Friday, October 26, 2007 1:07 PM
Subject: [Histonet] Eosinophil Staining

I am looking to do a comparative staining study on eosinophils (tissue
is in FFPE). Among several stains, I plan to include a
hematoxylin/eosin/azure II stain which will stain eosinophils pink
(Friend et al. 2000) and a Sirius red stain (Aust et al. 2000).
Unfortunately, for the H&E/azure II stain, no publications give the
protocol or specific stain sequence. As for the Sirius red stain, the
study states that the authors bought the stain from Bayer AG in Germany.
I have contacted Bayer and no one seems to know what I am talking about.
Before I substitute the "Bayer" Sirius red, I want to make sure that
using another company's stain is appropriate as the protocol is
described as "Sirius red (500mg) was dissolved in 45 ml aqua bidest, 50
ml absolute ethanol and 1 ml 1% NaOH; 4ml NaCl at 20% solution was added
until slight precipitation occurred." My main concern is that the Sirius
red I can order from other company's state there is little to no
solubility in ethanol. I am hoping someone will be able to help me in
regards to these questions. Thank you for your time.

Michelle Griffin-Reyes
Comparative Pathology Laboratory
University of Iowa Hospitals and Clinics
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