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Tissue clearing and three-dimensional imaging of the whole cochlea and vestibular system from multiple large-animal models.

ABSTRACT: The inner ear of humans and large animals is embedded in a thick and dense bone that makes dissection challenging. Here, we present a protocol that enables three-dimensional (3D) characterization of intact inner ears from large-animal models. We describe steps for decalcifying bone, using solvents to remove color and lipids, and imaging tissues in 3D using confocal and light sheet microscopy. We then detail a pipeline to count hair cells in antibody-stained and 3D imaged cochleae using open-source software. For complete details on the use and execution of this protocol, please refer to (Moatti et al., 2022).¹.

SUBMITTER: Moatti A

PROVIDER: S-EPMC10140170 | biostudies-literature | 2023 Apr

REPOSITORIES: biostudies-literature

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Tissue clearing and three-dimensional imaging of the whole cochlea and vestibular system from multiple large-animal models.

Moatti Adele A Cai Yuheng Y Li Chen C Popowski Kristen D KD Cheng Ke K Ligler Frances S FS Greenbaum Alon A

STAR protocols 20230413 2

The inner ear of humans and large animals is embedded in a thick and dense bone that makes dissection challenging. Here, we present a protocol that enables three-dimensional (3D) characterization of intact inner ears from large-animal models. We describe steps for decalcifying bone, using solvents to remove color and lipids, and imaging tissues in 3D using confocal and light sheet microscopy. We then detail a pipeline to count hair cells in antibody-stained and 3D imaged cochleae using open-sour ...[more]

PMID: 37060559

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