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

Biotechnology and Bioengineering Microscopy

Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
13 Apr 2023
Historique:
received: 09 11 2022
revised: 27 01 2023
accepted: 13 03 2023
entrez: 15 4 2023
pubmed: 16 4 2023
medline: 16 4 2023
Statut: aheadofprint

Résumé

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).

Identifiants

DOI: 10.1016/j.xpro.2023.102220 PMID: 37060559 PMC: PMC10140170
pubmed: 37060559
pii: S2666-1667(23)00178-8
doi: 10.1016/j.xpro.2023.102220
pmc: PMC10140170
pii:
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

102220

Subventions

Organisme : NIDCD NIH HHS
ID : K99 DC019960
Pays : United States
Organisme : NIDCD NIH HHS
ID : R21 DC020005
Pays : United States

Informations de copyright

Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.

Déclaration de conflit d'intérêts

Declaration of interests The authors declare no competing interests.

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Auteurs

Adele Moatti (A)

Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA; Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA.

Yuheng Cai (Y)

Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA; Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA.

Chen Li (C)

Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA; Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA.

Kristen D Popowski (KD)

Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA; College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27606, USA.

Ke Cheng (K)

Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA; Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA; College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27606, USA.

Frances S Ligler (FS)

Department of Biomedical Engineering, Texas A&M University, College Station, TX 77843, USA.

Alon Greenbaum (A)

Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27606, USA; Comparative Medicine Institute, North Carolina State University, Raleigh, NC 27606, USA. Electronic address: greenbaum@ncsu.edu.

Classifications MeSH