Format

Send to

Choose Destination
Hear Res. 2008 Sep;243(1-2):1-10. doi: 10.1016/j.heares.2008.05.009. Epub 2008 Jun 6.

Three-dimensional imaging of the intact mouse cochlea by fluorescent laser scanning confocal microscopy.

Author information

1
Virginia Merrill Bloedel Hearing Research Center, Department of Otolaryngology-HNS, University of Washington, Seattle, WA 98195, USA.

Abstract

The complex anatomy of the mammalian cochlea is most readily understood by representation in three-dimensions. However, the cochlea is often sectioned to minimize the effects of its anatomic complexity and optical properties on image acquisition by light microscopy. We have found that optical aberrations present in the decalcified cochlea can be greatly reduced by dehydration through graded ethanols followed by clearing with a mixture of five parts methyl salicylate and three parts benzyl benzoate (MSBB). Clearing the cochlea with MSBB enables acquisition of high-resolution images with multiple fluorescent labels, through the full volume of the cochlea by laser scanning confocal microscopy. The resulting images are readily applicable to three-dimensional morphometric analysis and volumetric visualizations. This method promises to be particularly useful for three-dimensional characterization of anatomy, innervation and expression of genes or proteins in the many new animal models of hearing and balance generated by genetic manipulation. Furthermore, the MSBB is compatible with most non-protein fluorophores used for histological labeling, and may be removed with traditional transitional solvents to allow subsequent epoxy embedding for sectioning.

PMID:
18573326
PMCID:
PMC2566306
DOI:
10.1016/j.heares.2008.05.009
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for Elsevier Science Icon for PubMed Central
Loading ...
Support Center