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.
Counterstaining for immunohistochemistry or other special stains.
- Sections to distilled water after IHC.
- Stain in methyl green solution for 5 minutes at room temperature (60 C may produce slightly stronger stain).
- Rinse in distilled water (sections will look blue).
- 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).
- Clear in xylene or xylene substitute.
- Mount with resinous mounting medium.
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