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

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

Author information

1
Tübingen Hearing Research Centre, Department of Otolaryngology, Head and Neck Surgery, Eberhard Karls University Tübingen, Tübingen, Germany; Department of Otorhinolaryngology and Head and Neck Surgery, University of Pécs, Pécs, Hungary.
2
Tübingen Hearing Research Centre, Department of Otolaryngology, Head and Neck Surgery, Eberhard Karls University Tübingen, Tübingen, Germany.
3
Tübingen Hearing Research Centre, Department of Otolaryngology, Head and Neck Surgery, Eberhard Karls University Tübingen, Tübingen, Germany; Massachusetts Eye and Ear Infirmary, Eaton-Peabody Laboratories, Harvard Medical School, Boston, MA, USA.
4
Department of Otorhinolaryngology and Head and Neck Surgery, University of Pécs, Pécs, Hungary.
5
School of Medicine and Health Sciences - European Medical School, University Hospital of Otorhinolaryngology, Medical Campus, University of Oldenburg, Steinweg 13-17, 26122 Oldenburg, Germany; Research Center of Neurosensory Science, University of Oldenburg, 26111 Oldenburg, Germany; Cluster of Excellence Hearing4all, University of Oldenburg, 26111 Oldenburg, Germany.
6
Tübingen Hearing Research Centre, Department of Otolaryngology, Head and Neck Surgery, Eberhard Karls University Tübingen, Tübingen, Germany; School of Medicine and Health Sciences - European Medical School, University Hospital of Otorhinolaryngology, Medical Campus, University of Oldenburg, Steinweg 13-17, 26122 Oldenburg, Germany; Research Center of Neurosensory Science, University of Oldenburg, 26111 Oldenburg, Germany; Cluster of Excellence Hearing4all, University of Oldenburg, 26111 Oldenburg, Germany. Electronic address: marcus.mueller@uni-tuebingen.de.

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

PMID:
26226655
DOI:
10.1016/j.jneumeth.2015.07.017
[Indexed for MEDLINE]
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