title: Decalcification by ascorbic acid for immuno- and affinohistochemical techniques on the inner ear.
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 degrees 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 (gamma-aminobutyric acid and calcitonin gene-related peptide), neurofilaments, and the galectine endogenous lectin were successfully detected immunohistochemically.
authors.affiliation: Center for Cell Culture (CAI-UCM) and Department of Surgery II (ORL), School of Medicine, Complutense University, Madrid, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Merchán-Pérez
authors.forename: P
authors.initials: P
authors.lastname: Gil-Loyzaga
authors.forename: M V
authors.initials: MV
authors.lastname: Bartolomé
authors.forename: M
authors.initials: M
authors.lastname: Remezal
authors.forename: P
authors.initials: P
authors.lastname: Fernández
authors.forename: T
authors.initials: T
authors.lastname: Rodríguez
mesh_terms: D000818:Animals; D001205:Ascorbic Acid; D018775:Bone Demineralization Technique; D002113:Calcification, Physiologic; D015740:Calcitonin Gene-Related Peptide / Q000032:analysis; D002795:Choline O-Acetyltransferase / Q000032:analysis; D003051:Cochlea / Q000737:chemistry / Q000473:pathology*; D006199:Hair Cells, Auditory, Inner / Q000737:chemistry; D018072:Hair Cells, Auditory, Outer / Q000737:chemistry; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D016900:Neurofilament Proteins / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D005680:gamma-Aminobutyric Acid / Q000032:analysis
chemical_list: D016900:Neurofilament Proteins; D005680:gamma-Aminobutyric Acid; D002795:Choline O-Acetyltransferase; D015740:Calcitonin Gene-Related Peptide; D001205:Ascorbic Acid