Microwave decalcification of human temporal bones.
OBJECTIVE
Morphological and immunohistochemical studies of human temporal bones are challenging as a result of difficulties in obtaining reliably fixed 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 examination revealed no obvious artifacts attributable to the microwave decalcification process. The quality of morphological preservation was largely dependent on the postmortem fixation 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 examination. Decalcification time is significantly reduced with no apparent adverse effects on structural preservation or antigenicity.
Cunningham
C D
CD
Department of Otolaryngology--Head and Neck Surgery, Medical University of South Carolina, Charleston 29425, USA.
Schulte
B A
BA
Bianchi
L M
LM
Weber
P C
PC
Schmiedt
B N
BN
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D003648:Decalcification Technique; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008872:Microwaves; D008875:Middle Aged; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013701:Temporal Bone / Q000473:pathology*; D016707:Tissue Fixation
D000254:Sodium-Potassium-Exchanging ATPase