High-resolution Immunofluorescence Imaging of Mouse Cochlear Hair Bundles

Overview

Journal STAR Protoc

Publisher Cell Press

Specialties Biology
Biomedical Engineering
Science

Date 2022 Jun 7

PMID 35669049

Authors

Katharine K Miller

Pei Wang

Nicolas Grillet

Affiliations

Soon will be listed here.

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).

Citing Articles

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LOXHD1 is indispensable for maintaining TMC1 auditory mechanosensitive channels at the site of force transmission.

Wang P, Miller K, He E, Dhawan S, Cunningham C, Grillet N Nat Commun. 2024; 15(1):7865.

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SUB-immunogold-SEM reveals nanoscale distribution of submembranous epitopes.

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VASCilia (Vision Analysis StereoCilia): A Napari Plugin for Deep Learning-Based 3D Analysis of Cochlear Hair Cell Stereocilia Bundles.

Kassim Y, Rosenberg D, Das S, Huang Z, Rahman S, Shammaa I bioRxiv. 2024; .

PMID: 38948743 PMC: 11212889. DOI: 10.1101/2024.06.17.599381.

SUB-Immunogold-SEM reveals nanoscale distribution of submembranous epitopes.

Miller K, Wang P, Grillet N Res Sq. 2024; .

PMID: 38343799 PMC: 10854333. DOI: 10.21203/rs.3.rs-3876898/v1.

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