DEMINERALIZATION OF BONE BY AMMONIUM-CHLORIDE FOR THE PYROANTIMONATE METHOD

Rat bones were fixed by the pyroantimonate method and demineralized by treatment with ammonium chloride (NH2Cl). Bones from very young rats (neonatal and a-week-old rats) that had been treated with NH4Cl were easy to cut into ultrathin sections. However, bones of 8-week-old rats needed to be demineralized for a longer time and were not very easy to cut. The extent of demineralization was measured by scintillation counting. After demineralization, precipitates of calcium pyroantimonate (Ca-pyroantimonate) were well retained and the fine structure of the tissue was well preserved. In addition, the distribution of Ca-pyroantimonate in the demineralized bone was similar to that in undecalcified specimens. This demineralization method is used in combination with the pyroantimonate method.