Experimental Studies in Demineralization and Its Effects on Cytology and Staining of Bone and Marrow Cells

The three types of bone used in this study were rib, iliac crest, and femoral head. 315 bone samples were variously fixed, demineralized, and histologically evaluated after paraffin embedment. Fixation in 10% buffered formalin followed by demineralization proved to be the best procedure for preservation of cellular and morphologic detail. If bone samples were to be demineralized in ethylene diaminetetraacetic acid disodium salt (EDTA), 70% ethanol fixation was better than formalin. Formalin fixation before EDTA caused hardness of bone more than alcohol fixation did, making it difficult to determine the end point of demineralization. No reagent was the ideal reagent for removing mineral salts from bone.Many of the procedures were standardized so the entire process could be timed as one block of activity. An empirical formula for demineralizing bone was proposed; and a reservoir of data as a baseline for monitoring quality control in the laboratory was developed; and the latest microtechniques for bone pathology were examined.