Rapid Method for localizing Beta-glucuronidase in Populations of Human Leucocytes and of Mouse Ehrlich Carcinoma Cells

Abstract

THE localization of beta-glucuronidase in populations of free cells has already been reported¹ from this laboratory. Thus, formaldehyde–chloral hydrate fixation of the sedimented cells followed by embedding in Knox gelatine¹ made it possible to cut thin sections of the frozen gelatine block on the freezing microtome and the sections so obtained were incubated while floating freely in Fishman–Baker substrate medium containing 8-hydroxyquinoline β-D-glucosiduronic acid and ferric ions. The enzymatically liberated 8-hydroxyquinoline became insoluble as the brownish-black ferric chelate at the sites rich in enzyme, and the ion was later converted to Prussian blue. The whole process required a minimum of 24 h from beginning to end and included many steps². Efforts to use dried smears with this technique were usually unsuccessful (except for one report)³ because of loss of cells from the slide during the various steps in the staining process.