A quantitative measure of the affinity of a dye for a substrate is given by: Affinity =
(where R is the gas constant, T the absolute temperature, C d the concentration of the dyebath). Affinities have been measured for several tissue substrates with a range of basis and “mordant” dyes.
The results obtained have been used to study the mechanism of several staining procedures of special interest. For example, the Critical Electrolyte Concentration effects could not, contrary to previous suggestions, be correlated with differential dye-substrate affinities in the salt-free situation. Another application was the study of methyl green-pyronin type stains for differential staining of DNA and RNA. The results failed to support the theories requiring specific dye-nucleic acid affinities. Finally an attempt was made to investigate the mode of action of “mordant” dyes of the preformed metal complex type. The results suggested that some dyes of these at least do not bind via the metal ion.
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Marshall, P.N., Horobin, R.W. Measurements of the affinities of basic and “mordant” dyes for various tissue substrates. Histochemie 36, 303–312 (1973). https://doi.org/10.1007/BF00305709