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.
Cunningham, C.D., B.A. Schulte, L.M. Bianchi, P.C. Weber, and B.N. Schmiedt. 2001. "Microwave decalcification of human temporal bones." Laryngoscope 111(2): 278-282.
John Wiley & Sons
Cochlea, Inner ear, Na, K-ATPase, Immunohistochemistry, Morphology