Decalcification by ascorbic acid for immuno- and affinohistochemical techniques on the inner ear |
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Authors: | Angel Merchán-Pérez P Gil-Loyzaga Maria Visitación Bartolomé Manuel Remezal Pilar Fernández Teresa Rodríguez |
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Institution: | Center for Cell Culture (CAI-UCM) and Department of Surgery II (ORL), School of Medicine, Complutense University, Madrid, Spain, ES
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Abstract: | An ascorbic acid decalcifying solution was applied to immuno- and affinohistochemical studies on the inner ear. Rat inner
ears fixed in 4% paraformaldehyde in PBS or in 2% acetic acid in ethanol solutions were adequately decalcified in an ascorbic
acid solution, at a temperature of 4°C. The decalcifying solution was prepared with 1% ascorbic acid and 0.84% sodium chloride
in distilled water (pH 2.5–2.6). The decalcification time was in a direct relationship to the specimen calcification. In this
study, two neuroactive substances (γ-aminobutyric acid and calcitonin gene-related peptide), neurofilaments, and the galectine
endogenous lectin were successfully detected immunohistochemically.
Accepted: 20 May 1999 |
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