Microwave decalcification of human temporal bones

Objectives/Hypothesis Morphological and immunohistochemical studies of human temporal bones are challenging as a result of difficulties in obtaining reliably fi-ed specimens and the lengthy time required for decalcification, typically 4 to 7 months. A novel method of processing human temporal bones using a microwave oven to accelerate decalcification is described. This procedure provides a rapid means of decalcifying temporal bones with good preservation of tissue morphology and antigenicity. Methods Human temporal bone specimens obtained at autopsy (n = 12, from specimens aged 43–91 y) were fixed within 6.5 hours of death by transtympanic perilymphatic perfusion of the inner ear. Decalcification was carried out using ethylenediaminetetra-acetic acid (EDTA) in a microwave oven and required only 3 to 6 weeks. Specimens were then dehydrated, embedded in paraffin, sectioned, and mounted on slides for morphological and immunohistochemical evaluation. Results Microscopic e-amination revealed no obvious artifacts attributable to the microwave decalcification process. The quality of morphological preservation was largely dependent on the postmortem fi-ation interval and adequacy of perilymphatic perfusion. Immunohistochemical analysis demonstrated strong positive staining for the enzyme Na,K-ATPase, an integral membrane protein. Conclusions This study demonstrates that microwave decalcification provides an efficient and reliable means of processing human temporal bones for histological and histochemical e-amination. Decalcification time is significantly reduced with no apparent adverse effects on structural preservation or antigenicity