A NEW METHOD FOR THE INVESTIGATION OF THE CAPTURE REACTION IN PHOSPHATASE CYTOCHEMISTRY III. EFFECTS OF THE COMPOSITION OF THE INCUBATION MEDIUM ON THE TRAPPING OF PHOSPHATE IONS IN A MODEL SYSTEM

Abstract

A model system developed for the study of diffusion problems in lead salt enzyme cytochemistry served as a basis for the experiments reported in the present paper. Phosphate leakage, which was investigated in polyacrylamide films during incubation in media containing lead, could be expressed by a graphical parameter related to the average displacement of the phosphate ions prior to the onset of lead phosphate precipitation in the films. This parameter can be used as a measure for the efficiency of the capture reaction in cytochemical reactions in tissue sections. The concentration of lead ions, and to a lesser extent that of the phosphate ions, was found to be critical for the degree of phosphate diffusion in this system. Addition of 0.3% of β-glycerophosphate to the lead medium gave increased phosphate diffusion, and trapping was markedly improved by added chloride ions. Raising of the molarity of the acetate buffer resulted in impaired trapping efficiency. Similar findings were made in a parallel study on the kinetics of lead phosphate precipitation in supersaturated solutions with a nephelometric method. The results of the present investigation indicate that the film model system is highly suitable for the study of factors affecting the rapid immobilization of diffusible products of cytochemical reactions.