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J Vis Exp. 2016 Jan 1;(107). doi: 10.3791/53561.

Whole Mount Dissection and Immunofluorescence of the Adult Mouse Cochlea.

Author information

1
Department of Surgery, Division of Otolaryngology, Southern Illinois University, School of Medicine.
2
Department of Surgery, Division of Otolaryngology, Southern Illinois University, School of Medicine; Department of Pharmacology, Southern Illinois University, School of Medicine; bcox@siumed.edu.

Abstract

The organ of Corti, housed in the cochlea of the inner ear, contains mechanosensory hair cells and surrounding supporting cells which are organized in a spiral shape and have a tonotopic gradient for sound detection. The mouse cochlea is approximately 6 mm long and often divided into three turns (apex, middle, and base) for analysis. To investigate cell loss, cell division, or mosaic gene expression, the whole mount or surface preparation of the cochlea is useful. This dissection method allows visualization of all cells within the organ of Corti when combined with immunostaining and confocal microscopy to image cells at different planes in the z-axis. Multiple optical cross-sections can also be obtained from these z-stack images. In addition, the whole mount dissection method can be used for scanning electron microscopy, although a different fixation method is needed. Here, we present a method to isolate the organ of Corti as three intact cochlear turns (apex, middle, and base). This method can be used for mice ranging from one week of age through adulthood and differs from the technique used for neonatal samples where calcification of the cochlea is incomplete. A slightly modified version can be used for dissection of the rat cochlea. We also demonstrate a procedure for immunostaining with fluorescently tagged antibodies.

PMID:
26779585
PMCID:
PMC4780967
DOI:
10.3791/53561
[Indexed for MEDLINE]
Free PMC Article

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