Selected area electron diffraction and high spatial resolution, nondispersive electron probe Xray microanalysis have been used to examine the nature of the solid phase mineral deposits of specific ultrastructural components in freshly dissected, undecalcified bone tissue from embryonic chicks, prepared anhydrously with either 100% ethylene glycol or dry ultracryomicrotomy. No electron diffraction patterns of a specific calcium phosphate solid phase are generated from the dense mitochondrial granules of osteoblasts, shown to contain calcium and phosphorus by electron probe microanalysis, and from the early mineral deposits from certain regions of newly synthesized bone. Similar results are obtained from a synthetic preparation of an amorphous calcium phosphate. Absence of an electron diffraction pattern is not caused by an insufficient mass of the calcium phosphate solid phase. The electron diffraction patterns of the more heavily mineralized older regions of the bone show the reflections and characteristics of poorly crystalline hydroxyapatite. There is a progressive change in the electron diffraction pattern approaching that of crystalline hydroxyapatite with increasing distance from the periosteal surface. Electron probe microanalysis of the same tissue components and regions shows that the changes in the electron diffraction characteristics are accompanied by an increase in the Ca/P ratios of the solid mineral phase.
Read full abstract