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

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

STAR protocols (2021-05-25)
Yan Lu, Fangfang Wang, Haoyu Wang, Philipp Bastians, Yunfeng Hua
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).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Spurr Low Viscosity Embedding Kit, hydrophobic resin for embedding histological samples
Sigma-Aldrich
Potassium hexacyanoferrate(II) trihydrate, ACS reagent, 98.5-102.0%
Sigma-Aldrich
Sodium cacodylate trihydrate, ≥98%