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High-resolution immunofluorescence imaging of mouse cochlear hair bundles.

ABSTRACT: High-resolution immunofluorescence imaging of cochlear hair bundles faces many challenges due to the hair bundle's small dimensions, fragile nature, and complex organization. Here, we describe an optimized protocol for hair-bundle protein immunostaining and localization. We detail the steps and solutions for extracting and fixing the mouse inner ear and for dissecting the organ of Corti. We further emphasize the optimal permeabilization, blocking, staining, and mounting conditions as well as the parameters for high-resolution microscopy imaging. For complete details on the use and execution of this protocol, please refer to Trouillet et al. (2021).

SUBMITTER: Miller KK

PROVIDER: S-EPMC9163841 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature

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High-resolution immunofluorescence imaging of mouse cochlear hair bundles.

Miller Katharine K KK Wang Pei P Grillet Nicolas N

STAR protocols 20220530 2

High-resolution immunofluorescence imaging of cochlear hair bundles faces many challenges due to the hair bundle's small dimensions, fragile nature, and complex organization. Here, we describe an optimized protocol for hair-bundle protein immunostaining and localization. We detail the steps and solutions for extracting and fixing the mouse inner ear and for dissecting the organ of Corti. We further emphasize the optimal permeabilization, blocking, staining, and mounting conditions as well as the ...[more]

PMID: 35669049

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