There is methyl violet in methyl green that needs to be removed using
chloroform and a separatory funnel.  It's a mess to do.  I don't remember
where I found the protocol for this, but I'm sure others will be able to
provide that.

We get around it by buying it commercially prepared.

-Teri Johnson
Physicians Reference Laboratory
Overland Park, KS

----- Original Message -----
From: kwalters <kwalters@blue.weeg.uiowa.edu>
To: histo <histonet@pathology.swmed.edu>
Sent: Thursday, April 12, 2001 9:45 AM
Subject: methyl green is methyl blue


> Everytime I make methyl green it turns a gorgeous shade of blue.  I have a
> brand new powder that I am using.
>
> Here is my protocol:  please tell me where I went wrong!!
>
> Thanks for any help!!
>
> Methyl Green
> a ) 0.1 N Acetic Acid: Add 6 ml of glacial acetic acid to 1000 ml of
>          filtered distilled water.
>
> b) 0.1 N sodium Acetate: Add 4.102 g of sodium acetate to 500 ml of
distilled
> water.
>
>          c) Buffer:
> 1)Add 755 ml of 0.1 N acetic acid to 265 ml of 0.1 N sodium
> acetate.
> 2) Adjust pH to 4.2 with NAOH.
>
>          d) Methyl Green
> 1) Add 20.0 g of methyl green to the buffer solution.
>          2) 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.
>
> Kathy Walters                                //
> Central Microscopy Research Facility        / /
> 85 EMRB                                    / /\
> University of Iowa                        / /\ \
> Iowa City, Iowa 52242                    / /  \ \
> Phone #:  (319) 335-8142                / /    \ \
> Fax #:  (319) 384-4469                ______   ((0))
> email: Katherine-Walters@uiowa.edu     |__|    / /
>                                         ||    / /
>                                      --------------
>                                    ------------------
>
>
>