Re: chromotrope 2R for eosinophil granules

From:Amy Porter <>

Cindy - we have used Lendrum's Method for staining
eosinophil polymorphonuclear leucocytes in our
laboratory just for that specific cell type.  I don't
know if it would interfere with your immuno reaction
or not.  Here it is in a nutshell:
Phenol crystals(pure)           1 gram
Chromotrope 2R                0.5 grams

Phenol crystals are placed in a flask and dissolved by
rotating under the hot tap water.  Next add the
chromotrope 2R and mix well.  The mixture is then
dissolved in 100.0 ml of distilled water and filtered.
 The stain is good for at least three months.

To Stain:  
1.  Sections are stained in the usual manner with an
alum haematoxylin, decolorized in 1% acid alcohol and
2.  Stain with Carbol-chromotrope for 30 - 60 minutes.
3.  Rinse in tap water.
4.  Dehydrate in the usual manner.
5.  Clear in Xylene.
6.  Mount in balsam or D.P.X.  (we use permount)

Eosinophil granules - bright red
Cell nuclei - blue
Red blood corpuscles - bright red

(J. Path. Bact.(1944) 56, 3.)  In a very old
manual-Practical Section Cutting and Staining fifth
edition by E.C. Clayden.

Hope this helps you out.
--- Cindy Burchell <> wrote:
> 	Hello Histonetters,
>     It's been a while since I have had the chance to
> be in contact with 
> you, and now I need a favor again.  My bosses insist
> that I use something 
> called Chromotrope 2R to counterstain eosinophilic
> granules after IHC 
> staining MBP of eos.  I've found references, but
> never any specific 
> methods.  I would be in debt to anyone who could
> spell out just how to 
> handle this counter stain.   Thanks so much in
> advance for your time if you 
> will help me in this matter!!!   Cindy B.

Amy S.Porter, HT(ASCP)
Michigan State University
College of Human Medicine

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