Decalcification by ascorbic acid for immuno- and affinohistochemical techniques on the inner ear.
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.
Merchán-Pérez
A
A
Center for Cell Culture (CAI-UCM) and Department of Surgery II (ORL), School of Medicine, Complutense University, Madrid, Spain.
Gil-Loyzaga
P
P
Bartolomé
M V
MV
Remezal
M
M
Fernández
P
P
Rodríguez
T
T
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
D016900:Neurofilament Proteins; D005680:gamma-Aminobutyric Acid; D002795:Choline O-Acetyltransferase; D015740:Calcitonin Gene-Related Peptide; D001205:Ascorbic Acid