Don't recall seeing the question posted but here it is:

Please note that this is not MY procedure but I do not remember where I got it.

Methyl Green Counterstain

REAGENTS:
a)	Acetic Acid, Glacial (aldehyde-free) 
b)	Sodium Acetate, Anhydrous 
c)	Methyl Green

MIX

1 )    0.1 N Acetic Acid: Add 6ml of glacial acetic acid to 1000 ml of filtered
distilled water.  

2)	0.1 N sodium Acetate: Add 4.102 g of sodium acetate to 500 ml of  distilled water.

 
  3)      Buffer:
   a) Add 755 ml of 0.1 N acetic acid to 265 ml of 0.1 N sodium acetate.
b)	Adjust pH to 4.2 with NAOH.

4)	Methyl Green 
a)	Add 20.0 g of methyl green to the buffer solution.
b)	Filter the methyl green solution.

STORAGE: room temperature
SHELF LIFE: Indefinite while in storage; when in use, the shelf life is 2 weeks or
until staining becomes too weak.

PROCEDURE:
1)	After color development of slides, rinse in distilled water briefly.

2)	Place slides in methyl green solution for 3 to 5 minutes, or as long as necessary
to get the
         staining desired. (slides which have been denatured, i.e. as for
in-situ-hybridization, 
         typically need far less Counterstain 30 seconds or so.)

3)	Do not rinse in water.  The methyl green washes out too easily.  Go directly into
95%  
          alcohol for a brief time (about 10 dips) .

4)	Continue dehydrating through one more 9S% alcohol, then 3 changes of 100% alcohol,
  
         allowing the slides to stay in the last alcohol for 2-3 minutes.

  5)	     Clear through 3-4 changes of xylene, and coverslip.


best regards,
Bob
Robert Schoonhoven
Laboratory of Molecular Carcinogenesis and Mutagenesis
Dept. of Environmental Sciences and Engineering
University of North Carolina
CB#7400
Chapel Hill, NC 27599
Phone 
office 919-966-6343
   Lab 919-966-6140
   Fax 919-966-6123 

Don't go around saying the world owes you a living; the world owes you
nothing; it was here first. 
Mark Twain [Samuel Langhornne Clemens] (1835-1910) 

-- Begin original message --

> From: Gayle Callis <uvsgc@msu.oscs.montana.edu>
> Date: Mon, 13 Sep 1999 22:37:15 -0600
> Subject: Bob Schoonhoven, dig out your methyl green counterstain again
> To: histonet@pathology.swmed.edu
> 
> Bob Schoonhoven has a methyl green counterstain that works very nicely, of course
> I have volunteered his expertise to come forth with the recipe/method.
> 
> Sorry, I am at home without my manual here, can't recite recipe
> 
> Gayle Callis
> 
> 

-- End original message --