Calcium Localized in Odontogenic Cells of Rat Mandibular Teeth by the Glyoxal Bis (2-Hydroxyanil) Method

¹ Department of Anatomy, University of Louisville School of Medicine and School of Dentistry, Louisville, Kentucky

The glyoxal bis(2-hydroxyanil) (GBHA) method was used to stain calcium in odontogenic cells in molars from mandibles of suckling rats. Fresh blocks of mandibles were sectioned transversely approximately 1 mm. thick and immersed in the GBHA solution for 16 hours. The GBHA-stained blocks were dehydrated in ethanol, cleared in xylene, and embedded in paraffin (m.p. 65°-70°C.). Paraffin sections 7µ thick were affixed to albumenized slides by finger pressure, dried, immersed in solutions of Na₂CO₃ and KCN to insure specificity for calcium, and counterstained with methylene blue. The red Ca-GBHA granules were localized in cytoplasm of ameloblasts, cementoblasts, and in odontoblastic processes, but not in the apatite. The granules in the ameloblasts were sparse and fine compared to the abundant coarse granules in the cementoblasts. The granule size in the odontoblastic processes was similar to that found in the processes of osteocytes and osteoblasts. Difficulties in obtaining intact tissues in sections from paraffin blocks of GBHA-stained undecalcified teeth suggested a need for another synthetic embedding medium.

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