Quantitative spatial distributions of calcium, phosphorus, and sulfur in calcifying epiphysis by high resolution electron spectroscopic imaging

Abstract

Electron spectroscopic imaging, a new technique that permits the quantitative detection of the spatial distributions of atomic elements at high resolution, has been applied to the epiphyseal zone of hypertrophy in the mouse for the visualization of calcium, phosphorus, and sulfur. Longitudinally sectioned epiphyseal growth plates reveal a developmental sequence in the longitudinal septum leading from a noncalcified matrix to a calcified matrix. During the early stages of this transition, matrix granules containing highly localized concentrations of P (200-400 atoms/nm2) are found spatially separate from Ca-containing sites. These Ca localizations displayed a concentration range of 20-350 atoms/nm2 and a complete spatial overlap with sulfur. At these sites, S levels range from 10 to 200 atoms/nm2. At a later stage, and therefore more proximal to the zone of provisional calcification, the usual scattered, irregularly shaped mineral deposits are found. These sites contain a virtual superposition of Ca with both P and S. The Ca/P and Ca/S ratios of these mineral deposits are predominantly 1.0 with only minor, locally varying ratios present.