An Improved Stain for Fresh, Mineralized Bone Sections

Abstract

Fresh, unfixed, unembedded, mineralized bone sections were hand ground or cut on a thin sectioning machine to 75-100 microns thickness; stained in the "Osteochrome" for 90 min. (for quick diagnosis) or 48 hr. (for complete permeation of the sections). Then they are washed in 0.1% zephiran chloride, rinsed in distilled water, differentiated in alcoholic 0.01% acetic acid, dehydrated in ascending alcohols, cleared in xylol, and permanently mounted in Harleco"s mounting medium. The stain was a single pre-buffered powder dissolved in 70% reagent methanol. The staining reactions were consistent; osteoid seams stained 2 major colors red and green. The method was proved useful in routine diagnostic work with bone biopsies from cases of metabolic bone disease, especially the osteomalacias.