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Automated muscle fiber type population analysis with ImageJ of whole rat muscles using rapid myosin heavy chain immunohistochemistry
Author(s) -
Bergmeister Konstantin D.,
Gröger Marion,
Aman Martin,
Willensdorfer Anna,
ManzanoSzalai Krisztina,
Salminger Stefan,
Aszmann Oskar C.
Publication year - 2016
Publication title -
muscle and nerve
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.025
H-Index - 145
eISSN - 1097-4598
pISSN - 0148-639X
DOI - 10.1002/mus.25033
Subject(s) - myosin , biceps , immunohistochemistry , population , staining , skeletal muscle , anatomy , muscle fibre , fiber type , biology , pathology , biomedical engineering , medicine , biochemistry , environmental health
: 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. Methods : 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. Results : 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%. Conclusions : 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