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p-HYDROQUINONE AND p-BENZOQUINONE AS HISTOCHEMICAL REAGENTS I. A NEW TETRAZOLIUM METHOD FOR AMINO GROUPS, AND THE MECHANISM OF FORMAZAN STAINING OF PARAFFIN AND FRESH FROZEN SECTIONS

G. G. CARMICHAEL 1 and STEPHANIE T. K. MANDER 1

1 Institute of Medical and Veterinary Science, Adelaide, South Australia

The staining of amino groups by formazan when dehydrated paraffin sections are incubated in a mixture of hydroquinone and 3-(4,5-dimethyl thiazolyl-2)-2 ,5-diphenyl-2H-tetrazolium bromide at an acid pH is reported. The mechanism of this reaction and of the cytoplasmic deposition of formazan in fresh frozen sections incubated under similar conditions is investigated. It is shown that the oxidation of hydroquinone to semiquinone is responsible for the reaction, the tetrazole acting as electron acceptor. The tissue amino groups, exposed by dehydration and thermal shrinkage, and the nitrogen groupings of phosphobipid behave as "catalysts." The relevant properties of the hydroquinone-benzoquinone oxidation-reduction system are described, and the reactions between benzoquinone and tissue constituents are reviewed.

Submitted on January 23, 1967


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