The Nature of the Specificity of Methyl Green for Chromatin

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Abstract
Photometric cytological analysis demon-strates that methyl green (a basic dye) stains specifically for desoxypentose nucleic acid, that it is quantitatively related to the amt. of nucleic acid present and that it is dependent upon the physical state of the nucleic acid. When slides have been stained with purified dye in phenol-alcohol-glycerin soln. and decolorized in tertiary butyl alcohol to leave only chemically bound dye, the methyl green stains only chromatin. After removal of nucleic acid with trichloracetic acid there is no appreciable staining; the dye is therefore staining the nucleic acid, not the protein, of chromatin. Two hrs. action of ribonuclease on the sections has no effect upon methyl green staining; therefore methyl green is bound to desoxypentose nucleic acid, not to the pentose type. After treatment of sections with hot water there is loss of the staining property; this is ascribed to a physical change (depoly-merization) in the nucleic acid molecule. Desoxypentose nucleic acid is not removed from chromatin by hot water; the Feulgen reaction remains undiminished. The combined use of Feulgen reaction and methyl green staining is potentially an indicator of physical alteration of the desoxypentose nucleic acid molecule, for example, where chromatin is degenerating.

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