Re: methyl green counterstain that works

From:Lorraine Gibbs <>

I have never had a problem with Methyl Green. Here is the formula I use:

To 500mls of 0.1M acetate buffer, pH 4.2, add 10 grams of Methyl Green (CI#
42585). Stir well and filter. I usually filter before each use. Rinse in
DH20 pre and post stain. Staining times vary from 20sec-3 minutes. Tissue
should be a medium blue-green. Dehydrate quickly through ethanols (the
sections should now be a lighter green by the time they reach the final 100%
Clear & coverslip as usual.

Happy Staining!

Lorraine Gibbs
Physiology & Biophysics
University of Washington
Seattle WA
-----Original Message-----
From: Gayle Callis <>
To: <>
Date: Saturday, February 10, 2001 11:28 AM
Subject: methyl green counterstain that works

>Go into Histonet archives and look for Bob Schoonhoven's methyl green
>counterstain, it is gangbusters, made up in an acetate buffer, for IHC
>work.  Works with immunogold, enhanced, and should work with DAB or
>whatever chromogen combination with MG gives good contrast.
>Many pathologists like hematoxylin for the detail it provides (what they
>are used to looking at) morphologically.  You can dilute progressive
>hematoxylins with distilled water, but they tend to break down after this
>happens, but it will give delicate hematoxylin staining if you are bothered
>by heavy blues.
>Gayle Callis
>Veterinary Molecular Biology
>Montana State University - Bozeman
>Bozeman MT 59717-3610
>406 994-6367
>404 994-4303 (FAX)

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