Decalcification of Bone in Relation to Staining and Phosphatase Technics

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Abstract
The rate of decalcification of compact bone is increased by use of mineral acids, by increase in concn. of acids, by rise in temp. of mineral acid decalcifying baths. The accelerating effect of electrolysis on decalcification is due entirely to the rise in temp. occasioned thereby. Acid treatment at 58[degree]C rapidly decalcifies bone and then completely dissolves it. Alcohol and low temps. each retard decalcification. 1 [image] mineral acids dissolve about 6.5% Ca phosphate, acetic about 0.3% and formic about 2.5%. Formic acid decalcifies more slowly at 37[degree] than at 20[degree]C. Mineral acid decalcification affects Feulgen and azure eosin staining adversely, especially at higher temps., and formic acid at 37[degree]C also impairs these and Van Gieson stains. Hematoxylin eosin stains are less affected by these factors. For demonstration of alkaline phosphatase use of buffers of pH 4.5 or higher as decalcifying is required. Decalcification at 3[degree]C is recommended. Acetate or citrate buffers of 1 [image] concentration act more rapidly and preserve phosphatase as well as the weaker buffers hitherto used.
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