Protocol to image and analyze the morphology of mouse peripheral nerves using transmission electron microscopy

Hiromi Nakai-Shimoda; Tatsuhito Himeno; Mikio Motegi; Norio Ozeki; Rieko Inoue; Tomohide Hayami; Emiri Miura-Yura; Yuichiro Yamada; Yoshiaki Morishita; Shin Tsunekawa; Yoshiro Kato; Yusuke Seino; Masaki Kondo; Keiko Naruse; Koichi Kato; Hiroki Mizukami; Jiro Nakamura; Hideki Kamiya

doi:10.1016/j.xpro.2022.101591

Protocol to image and analyze the morphology of mouse peripheral nerves using transmission electron microscopy

STAR Protoc. 2022 Aug 1;3(3):101591. doi: 10.1016/j.xpro.2022.101591. eCollection 2022 Sep 16.

Authors

Hiromi Nakai-Shimoda¹, Tatsuhito Himeno², Mikio Motegi¹, Norio Ozeki¹, Rieko Inoue¹, Tomohide Hayami¹, Emiri Miura-Yura¹, Yuichiro Yamada¹, Yoshiaki Morishita¹, Shin Tsunekawa¹, Yoshiro Kato¹, Yusuke Seino³, Masaki Kondo¹, Keiko Naruse⁴, Koichi Kato⁵, Hiroki Mizukami⁶, Jiro Nakamura⁷, Hideki Kamiya¹

Affiliations

¹ Division of Diabetes, Department of Internal Medicine, Aichi Medical University School of Medicine, Nagakute 480-1185, Japan.
² Division of Diabetes, Department of Internal Medicine, Aichi Medical University School of Medicine, Nagakute 480-1185, Japan; Department of Innovative Diabetes Therapy, Aichi Medical University School of Medicine, Nagakute 480-1185, Japan. Electronic address: thimeno@aichi-med-u.ac.jp.
³ Division of Endocrinology and Metabolism, Department of Internal Medicine, Fujita Health University School of Medicine, Toyoake, Aichi 470-1192, Japan.
⁴ Department of Internal Medicine, Aichi Gakuin University School of Dentistry, Nagoya 464-0821, Japan.
⁵ Department of Medicine, Aichi Gakuin University School of Pharmacy, Nagoya 464-8650, Japan.
⁶ Department of Pathology and Molecular Medicine, Hirosaki University Graduate School of Medicine, Hirosaki 036-8562, Japan.
⁷ Division of Diabetes, Department of Internal Medicine, Aichi Medical University School of Medicine, Nagakute 480-1185, Japan; Department of Innovative Diabetes Therapy, Aichi Medical University School of Medicine, Nagakute 480-1185, Japan.

Abstract

Morphological analysis of peripheral nerves in mouse models can be used to characterize the pathophysiology of peripheral nerve disease, but obtaining high-quality electron micrographs can be challenging. Here, we present a protocol to obtain electron micrographs of mouse peripheral nerves. We detail the procedures of sampling, fixation, and embedding of peripheral nerves. We then outline the steps for ultrathin sectioning and transmission electron microscopy imaging. Finally, we describe morphological evaluation of nerve fibers in these images using ImageJ and AxonSeg. For complete details on the use and execution of this protocol, please refer to Nakai-Shimoda et al. (2021).

Keywords: Metabolism; Microscopy; Neuroscience.

Publication types

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

MeSH terms

Animals
Histological Techniques* / methods
Mice
Microscopy, Electron, Transmission
Peripheral Nerves* / diagnostic imaging
Specimen Handling