Large-scale 3D imaging of mouse cochlea using serial block-face scanning electron microscopy

Yan Lu; Fangfang Wang; Haoyu Wang; Philipp Bastians; Yunfeng Hua

doi:10.1016/j.xpro.2021.100515

Large-scale 3D imaging of mouse cochlea using serial block-face scanning electron microscopy

STAR Protoc. 2021 May 4;2(2):100515. doi: 10.1016/j.xpro.2021.100515. eCollection 2021 Jun 18.

Authors

Yan Lu¹, Fangfang Wang¹, Haoyu Wang¹, Philipp Bastians², Yunfeng Hua¹

Affiliations

¹ Shanghai Institute of Precision Medicine, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
² ZEISS Research Microscopy Solutions, Shanghai, China.

Abstract

This protocol describes how to prepare intact mouse cochleae for serial block-face scanning electron microscopy (SBEM). The detailed workflow includes cochlea fixation, en bloc staining, resin embedding, X-ray microscopy-guided trimming and SBEM data acquisition. This protocol allows large-scale, nanometer-resolution three-dimensional imaging of subcellular structures in a targeted tonotopic range of the cochlea and enables fast volumetric scan at submicron resolution using a compact X-ray microscope. For complete details on the use and execution of this protocol, please refer to Hua et al. (2021).

Keywords: Cell Biology; Microscopy; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cochlea / ultrastructure*
Imaging, Three-Dimensional*
Mice
Microscopy, Electron, Scanning*
Microtomy*