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Methyl Green Counterstain Protocol
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Methyl Green Counterstain Protocol


0.1M Sodium Acetate Buffer, pH4.2:

  • Sodium acetate, trihydrate (MW 136.1) ------ 1.36 g
  • Distilled water --------------------------------- 100 ml
  • Mix to dissolve and adjust pH to 4.2 using concentrated glacial acetic acid

Methyl Green Solution (0.5%)

  • Methyl green (ethyl violet free from Sigma) --- 0.5 g
  • 0.1M Sodium acetate buffer, pH4.2 ------------ 100 ml
  • Mix to dissolve.

Staining Pattern


Suggested Use

Counterstaining for immunohistochemistry or other special stains.

Staining Procedure

1.  Sections to distilled water after IHC.

2.  Stain in methyl green solution for 5 minutes at room temperature (60 C may produce slightly stronger stain).

3.  Rinse in distilled water (sections will look blue).

4.  Dehydrate quickly through 95% alcohol (10 dips, sections turn green), 2 changes of 100% alcohol (10 dips each) (alcohol used for dehydration removes some of the stain).

5.  Clear in xylene or xylene substitute.

6.  Mount with resinous mounting medium.

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