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HISTOCHEMICAL DEMONSTRATION OF CYTOCHROME OXIDASE WITH NEW AMINE REAGENTS
1 National Institute of Dental Research, National Institutes of Health, Bethesda 14, Maryland.
A series of aryl amines were employed as new histochemical reagents for the demonstration of cytochrome oxidase. The amines, upon oxidation, form indamine-azine type dyes which are capable of chelating with a number of metals.
The most useful substrate consisted of a mixture of N-phenyl-p-phenylenediamine (I) and p-methoxy-p-aminodiphenylamine (II) or 3-amino-9-ethyl-carbazole (V).
The reaction was inhibited by pretreatment of sections with saline, and augmented by the addition of cytochrome c. Heat, cyanide, and sulfide were inhibitory.
Although raw-frozen sections were employed, mitochondrial localizations were obtained in specific instances in selected areas of certain tissues.
The possibility of demonstrating other oxidative enzymes with these substrates warrants further study.
Submitted on June 2, 1959
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