Investigation of Thiazin Dyes as Biological Stains I. The Staining Properties of Thionin and its Derivatives as Compared with Their Chemical Formulae

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Abstract
An investigation has been made of the staining properties of eight dyes of the thionin group. The dyes studied are as follows: tetra-ethyl thionin, asymmetrical di-ethyl thionin, tetra-methyl thionin (methylene blue), tri-methyl thionin (azure B), asymmetrical di-methyl thionin (azure A), symmetrical di-methyl thionin, mono-methyl thionin (azure C), and unsubstituted thionin. The staining properties were tested on sections of paraffin embedded material following five different methods of fixation. No counterstain was employed. It was shown that there was a general correlation between the extent of ethylation or methylation of the dyes and their staining properties. As one passes from tetra-ethyl thionin down the series to thionin itself, there is a progressive decrease in the amount of green showing in the preparations, and an increase in the amount of red present, also an increase in the metachromatic effects, and in the intensity of nuclear staining. There seems, also, to be a similar relation between staining qualities on the one hand and the color and solubility of the dye base on the other.

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