A DIRECT CYTOCHEMICAL METHOD FOR THE IDENTIFICATION OF LYSOZYME IN VARIOUS TISSUES

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Abstract
This report describes a cytochemical technique for the demonstration of lysozyme in a variety of cells, including normal human and rabbit neutrophil granules, rabbit lacrimal gland cells and blasts obtained from bone marrow and peripheral blood of patients with myelomonocytic and histiomonocytic leukemia. It utilizes the dis-azo dye Biebrich scarlet which stains lysozyme as well as other basic proteins. The method is made specific for lysozyme by the use of N-acetylglucosamine oligosaccharides obtained from an acid hydrolysate of chitin. These greatly diminish the color reaction obtained when Biebrich scarlet is added to lysozyme, presumably by competing with Biebrich scarlet for the active site of the lysozyme molecule. Cytochemically, this is expressed as a marked diminution in the ability of Biebrich scarlet to stain the cytoplasm of lysozyme-containing cells if these cells have been pretreated with a hydrolysate of chitin. Evidence is presented that this stain is specific for lysozyme rather than a nonspecific reaction between Biebrich scarlet and tissue bases, particularly arginine.

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