Decalcification by ascorbic acid for immuno- and affinohistochemical techniques on the inner ear

Histochem Cell Biol. 1999 Aug;112(2):125-30. doi: 10.1007/s004180050398.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Ascorbic Acid*
  • Bone Demineralization Technique*
  • Calcification, Physiologic*
  • Calcitonin Gene-Related Peptide / analysis
  • Choline O-Acetyltransferase / analysis
  • Cochlea / chemistry
  • Cochlea / pathology*
  • Hair Cells, Auditory, Inner / chemistry
  • Hair Cells, Auditory, Outer / chemistry
  • Immunohistochemistry / methods
  • Male
  • Neurofilament Proteins / analysis
  • Rats
  • Rats, Sprague-Dawley
  • gamma-Aminobutyric Acid / analysis

Substances

  • Neurofilament Proteins
  • gamma-Aminobutyric Acid
  • Choline O-Acetyltransferase
  • Calcitonin Gene-Related Peptide
  • Ascorbic Acid