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NEW DIAZONIUM COMPONENTS AS COUPLING AGENTS IN THE DEMONSTRATION OF PHOSPHATASES

MARVIN S. BURSTONE 1 and ELIZABETH K. WEISBURGER 1

1 National Institute of Dental Research and National Cancer Institute, National Institutes of Health, Bethesda 14, Maryland

A histochemical technique for the diazotization of highly insoluble amines is described. The diazotization procedure is carried out in the presence of inert solvents, dimethylformamide or demethylsulfoxide.

Most of the amines contain coordinating groups which facilitate chelation of the final dyestuff with metals, including copper and cobalt. The aforementioned coordinating groups include nitrogen, keto, hydroxyl, amino, carboxyl, and the azo grouping. Several dye chelates were found to be highly resistant to xylene and ethanol.

In addition to the successful application of chelation techniques to azo-dye enzyme histochemistry, the first use of a metal containing diazonium salt is reported. The compound is diazotized p-(acetoxymercuri)-aniline. Diazotized 3-aminofluoranthene is highly stable in solution and is suitable for use in simultaneous coupling techniques.

Use of complex diazotized amines possessing coordinating groups for subsequent chelation enhances chromogenicity and fineness of localization and may find application in electron microscopy.

Submitted on August 3, 1960


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