HISTOCHEMICAL METHODS FOR ACID PHOSPHATASE USING HEXAZONIUM PARAROSANILIN AS COUPLER

pimatase activity is largely vitiated by the insperfections of the available histochenmical methods. The sources of error its the lead sulfide nmetimod weme appreciated by Gomori, but this ha.s usot discouraged the uncritical use of this technique in the area of “appbied” histochenmsistmy. ‘[he azo (lye nmethods have suffered fronms two deficiencies : a sui)strate that would yield a hydrolysis iroduct (If low soiubilitv and Imigh substantivity, amsd a capture reagent tisat would couple rapidly ins acid nmedia to productaum ussoluble anmsorphous azo dye. TIme pisos mhate esters of various usapimthob AS derivatives, as developed by Burstone (9), gave imimroved localizatious its freeze-dried material because of the increased l)roteius affinity aumci cxtrenumeby low’ solubibity of the naphtlmobic isycirolysis product. On the other sicica major refineummeust in diazomuiunms coupling tec-immmique followed time introduction of hexazonium b)ararosausilin by Davis and Ornstein (13). ‘[he use of freslsbv diazotizecl pararosanilin w-itim a-msaphthyb pisosphate its a sinmultaneous coupling azo dye method for acid phosphatase (4) resulted in significant improveumment because of the nmaumy desirable characteristics of the final azo dye (1, 2). Several logical msmodificatiomss that conmbimse time advantages of time naphthol AS sui)strates and the isexazonium pararosanibin coupler have givo-us reproducible artifact-free bocabizatioum of acid