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Proceedings Paper

Imaging living hair cells within the cochlear epithelium of mice using two-photon microscopy
Author(s): Tao Yuan; Simon S. Gao; Peter Saggau; John S. Oghalai
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Paper Abstract

Mice are an excellent model for studying mammalian hearing and transgenic mouse models of human hearing loss are commonly available for research. However, the mouse cochlea is substantially smaller than other animal models routinely used to study cochlear physiology. This makes the study of their hair cells difficult. We developed a novel methodology to optically image calcium within living hair cells left undisturbed within the excised mouse cochlea. Fresh cochleae were harvested, left intact within their otic capsule bone, and glued upright in a recording chamber. The bone overlying the region of the cochlear epithelium to be studied was opened and Reissner's membrane was incised. A fluorescent indicator was applied to the preparation to image intracellular calcium. A custom-built upright two-photon microscope was used to image the preparation using three dimensional scanning. We were able to image about 1/3 of a cochlear turn simultaneously, in either the apical or basal regions. Within one hour of animal sacrifice, we found that outer hair cells demonstrated increased fluorescence compared with surrounding supporting cells. Thus, this methodology can be used to visualize hair cell calcium changes and mechanotransduction over a region of the epithelium. Because the epithelium is left within the cochlea, dissection trauma is minimized and artifactual changes in hair cell physiology are reduced.

Paper Details

Date Published: 23 February 2009
PDF: 9 pages
Proc. SPIE 7182, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII, 718209 (23 February 2009); doi: 10.1117/12.809789
Show Author Affiliations
Tao Yuan, Baylor College of Medicine (United States)
Simon S. Gao, Rice Univ. (United States)
Peter Saggau, Rice Univ. (United States)
Baylor College of Medicine (United States)
John S. Oghalai, Baylor College of Medicine (United States)
Rice Univ. (United States)

Published in SPIE Proceedings Vol. 7182:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues VII
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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