Automated muscle fiber type population analysis with ImageJ of whole rat muscles using rapid myosin heavy chain immunohistochemistry

Muscle & Nerve
Konstantin D BergmeisterOskar C Aszmann

Abstract

Skeletal muscle consists of different fiber types which adapt to exercise, aging, disease, or trauma. Here we present a protocol for fast staining, automatic acquisition, and quantification of fiber populations with ImageJ. Biceps and lumbrical muscles were harvested from Sprague-Dawley rats. Quadruple immunohistochemical staining was performed on single sections using antibodies against myosin heavy chains and secondary fluorescent antibodies. Slides were scanned automatically with a slide scanner. Manual and automatic analyses were performed and compared statistically. The protocol provided rapid and reliable staining for automated image acquisition. Analyses between manual and automatic data indicated Pearson correlation coefficients for biceps of 0.645-0.841 and 0.564-0.673 for lumbrical muscles. Relative fiber populations were accurate to a degree of ± 4%. This protocol provides a reliable tool for quantification of muscle fiber populations. Using freely available software, it decreases the required time to analyze whole muscle sections. Muscle Nerve 54: 292-299, 2016.

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Citations

Jan 10, 2018·Proceedings of the National Academy of Sciences of the United States of America·Marcus D GoncalvesLewis C Cantley
Jul 11, 2020·Skeletal Muscle·Evi MasscheleinKatrien De Bock
Oct 7, 2017·Journal of Applied Physiology·Yuan WenKenneth S Campbell
Jan 8, 2019·Science Advances·Konstantin D BergmeisterOskar C Aszmann
Oct 21, 2020·Biomedicines·Laura C GunderDave Bridges
Oct 19, 2019·Environmental Toxicology and Pharmacology·Wenhuan YaoJiliang Si
Nov 18, 2020·Skeletal Muscle·Lyle W BabcockSusan L Hamilton
Feb 22, 2021·Ageing Research Reviews·Osvaldo DelbonoMaria Laura Messi

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