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

• Published in: STAR protocols Vol. 3; no. 3; p. 101591
• Main Authors: Nakai-Shimoda, Hiromi, Himeno, Tatsuhito, Motegi, Mikio, Ozeki, Norio, Inoue, Rieko, Hayami, Tomohide, Miura-Yura, Emiri, Yamada, Yuichiro, Morishita, Yoshiaki, Tsunekawa, Shin, Kato, Yoshiro, Seino, Yusuke, Kondo, Masaki, Naruse, Keiko, Kato, Koichi, Mizukami, Hiroki, Nakamura, Jiro, Kamiya, Hideki
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 16.09.2022; Elsevier
• Subjects: Metabolism; Microscopy; Neuroscience; Protocol; Microscopy; Metabolism; Neuroscience
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101591

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). [Display omitted] •Morphological analysis of mouse peripheral nerves using transmission electron microscopy•Step-by-step guide for sampling, fixation, and embedding of peripheral nerves•Ultrathin sectioning by glass knife and imaging using transmission electron microscopy•Image analysis of unmyelinated and myelinated nerve fibers using ImageJ and AxonSeg Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.