
==== Front
bioRxiv
BIORXIV
bioRxiv
2692-8205
Cold Spring Harbor Laboratory

10.1101/2024.08.22.608909
preprint
1
Article
Protocol for muscle fiber type and cross-sectional area analysis in cryosections of whole lower mouse hindlimbs
Battey Edmund http://orcid.org/0000-0003-4465-7533

Biswas Dipsikha http://orcid.org/0000-0003-0990-9678

Dos Santos Matthieu http://orcid.org/0000-0002-7113-102X

Maire Pascal http://orcid.org/0000-0001-7795-0029

Sakamoto Kei http://orcid.org/0000-0001-8839-5980

22 8 2024
2024.08.22.608909https://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License, which allows reusers to distribute, remix, adapt, and build upon the material in any medium or format, so long as attribution is given to the creator. The license allows for commercial use.
http://biorxiv.org/lookup/doi/10.1101/2024.08.22.608909
nihpp-2024.08.22.608909.pdf
We outline a robust, simple, and cost-effective method to simultaneously visualize all mouse lower hindlimb skeletal muscles. We describe procedures for orientating the whole lower hindlimb in gum tragacanth prior to freezing, simplifying the proceeding experimental steps, and enhancing the clarity and comprehensiveness of characterizations. We then detail steps for quantifying muscle fiber size and fiber type characteristics in a single cryosection using immunofluorescence and histochemistry. This protocol can be applicable for commonly used histological and (immuno) histochemical evaluations such muscle degeneration/regeneration, fibrosis, immune cell infiltration, enzymatic activity and glycogen content.
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pmc
