High-resolution immunofluorescence imaging of mouse cochlear hair bundles

Katharine K. Miller; Pei Wang; Nicolas Grillet

doi:10.1016/j.xpro.2022.101431

High-resolution immunofluorescence imaging of mouse cochlear hair bundles

Katharine K. Miller, Pei Wang, Nicolas Grillet

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

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).

Original language	English
Article number	101431
Journal	STAR Protocols
Volume	3
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2022.101431
State	Published - Jun 17 2022

Bibliographical note

Publisher Copyright:
© 2022 The Author(s)

Funding

The work was funded by the Department of Otolaryngology — Head & Neck Surgery startup funding and National Institute on Deafness and Other Communication Disorders (NIDCD) grants 1R21DC019457-01 and RO1-DC-016409-01A1 . We thank the Department of Otolaryngology — Head & Neck Surgery imaging core facility, in particular Patrick Atkinson, for input. We thank Shari and Kenneth Eberts, the Oberndorf family, and the SICHL contributors for their support. The authors would like to thank Enago ( www.enago.com ) for the English language review. License for the graphics of the ear (Item ID: 2051711780 ) was purchased from Shutterstock.com . The work was funded by the Department of Otolaryngology — Head & Neck Surgery startup funding and National Institute on Deafness and Other Communication Disorders (NIDCD) grants 1R21DC019457-01 and RO1-DC-016409-01A1. We thank the Department of Otolaryngology — Head & Neck Surgery imaging core facility, in particular Patrick Atkinson, for input. We thank Shari and Kenneth Eberts, the Oberndorf family, and the SICHL contributors for their support. The authors would like to thank Enago (www.enago.com) for the English language review. License for the graphics of the ear (Item ID: 2051711780) was purchased from Shutterstock.com. N.G. and K.K.M. formulated the project. K.K.M. and P.W. carried out methodology and investigation. N.G. and K.K.M. wrote the manuscript. The authors declare no competing interests.

Funders	Funder number
Yale Department of Otolaryngology
National Institute on Deafness and Other Communication Disorders	2051711780, R21DC019457, RO1-DC-016409-01A1

Keywords

Antibody
Cell Biology
Microscopy
Model Organisms
Neuroscience

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

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10.1016/j.xpro.2022.101431

Cite this

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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).",

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AB - 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).

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High-resolution immunofluorescence imaging of mouse cochlear hair bundles

Abstract

Bibliographical note

Funding

Keywords

ASJC Scopus subject areas

Access to Document

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Cite this