Methyl Green Counterstain Protocol


Methyl green is used commonly with bright-field microscopes to stain cell nuclei by dyeing the chromatin of cells  with a green color. It is chemically related to cresyl violet.


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)
  5. 2 changes of 100% alcohol (10 dips each) (alcohol used for dehydration removes some of the stain).
  6. Clear in xylene or xylene substitute.
  7. Mount with resinous mounting medium.


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