Tina ,

Heres the method I use and it works great

Mix solution of 0.5% (w/v) methyl green in 0.1M sodium acetate, pH to 4.0. 
(this solution can be kept for several months at room temperature and reused)

1)After the application of your substrate(s), wash sections with dH2O.
2)Immerse in methyl green for 10 min at room temperature.
3)Wash well with dH2O.
4)Immerse sections in 3 changes of 100% N-butanol -- dip rack 10 times in first and second and allow to sit for 30 sec in third.
5)Immerse sections in 3 changes of xylenes--2 min each.

Hope this helps,

angeline
dept. of neurology
medical college of georgia
amartin@mail.mcg.edu



>From: Tina Cardamone <kcard@myriad.its.unimelb.edu.au>
>Subject: methyl green as counterstain 

>Hello,
>
>I have a simple question regarding methyl green.
>Does anyone have a methyl green counterstain  protocol for 
>immunohistochemistry.
>It sounds simple however I can't seem to find a protocol anywhere.
>
>Thanks in advance.

>Tina Cardamone
>University of Melbourne.