PHOSPHORYLATION AS A HISTOCHEMICAL PROCEDURE

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Abstract
(1) The use of phosphoryl chloride (phosphorus oxychloride) in pyridine as a reagent for phosphorylation of tissue components is described. (2) Phosphorylation of many carbohydrate-containing substances is obtained by this method, including mucins, connective tissue (elastic tissue, reticulum, collagen, bone matrix), some glycolipids, glycogen and other tissue components. (3) Certain chemical entities that can be phosphorylated probably do not contain carbohydrate, but are lipids (either unsaturated or incompletely esterified), or proteins with free sulfhydryl, hydroxyl, or amino groups. (4) Phosphorylation reduces metachromasia of some tissue components (e.g. cartilage matrix), but enhances that of many others. The basophilia and metachromasia of phosphorylated components persist when staining is done in strongly acid solutions. (5) The significance of direct leukofuchsin positivity of many phosphorylated substances, and of relatively red staining of such substances with alum hematoxylin, and aldehyde-fuchsin, is not certain.