Methyl methacrylate embedding to study the morphology and immunohistochemistry of adult guinea pig and mouse cochleae

J Neurosci Methods. 2015 Oct 30;254:86-93. doi: 10.1016/j.jneumeth.2015.07.017. Epub 2015 Jul 29.

Abstract

Background: Histological analysis of the cochlea is required to understand the physiological and pathological processes in the inner ear. In the past, many embedding techniques have been tested in the cochlea to find an optimal protocol that gives both good morphological and immunohistochemical results. Resins provide high quality cochlear morphology with reduced immunogenicity due to the higher polymerization temperature.

New method: We used Technovit 9100 New(®), a low temperature embedding system based on methyl methacrylate, on adult guinea pig and mouse cochleae to evaluate preservation of the morphology and maintenance of the antigenicity.

Results: Conventional toluidine blue staining, as well as immunohistochemical staining with a set of commonly used antibodies, showed highly preserved morphology and immunogenicity of decalcified adult guinea pig and mouse cochleae.

Comparison with existing method(s): We demonstrate both, well-preserved morphology and preservation of antigenicity, superior to other embedding techniques.

Conclusions: Our results showed that the Technovit 9100 New(®) embedding system provided highly preserved morphology and immunogenicity with our protocol in adult guinea pig and mouse cochleae.

Keywords: Cochlea; Immunohistochemistry; Morphology; Organ of Corti; Resin embedding; Spiral ganglion neuron.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calbindin 2 / metabolism
  • Cochlea / cytology*
  • Cochlea / metabolism*
  • Cold Temperature
  • Coloring Agents
  • Epoxy Compounds
  • Guinea Pigs
  • Immunohistochemistry / methods*
  • Methylmethacrylate*
  • Mice
  • Tissue Embedding / methods*
  • Tolonium Chloride

Substances

  • Calbindin 2
  • Coloring Agents
  • Epoxy Compounds
  • Tolonium Chloride
  • Methylmethacrylate