ArticleJournal of visualized experiments : JoVE2017
A Rapid Automated Protocol for Muscle Fiber Population Analysis in Rat Muscle Cross Sections Using Myosin Heavy Chain Immunohistochemistry.
Article in Journal of visualized experiments : JoVE, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 12 citations in OpenAlex.
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- Mapping the functional anatomy and topography of the cardiac autonomic innervation for selective cardiac neuromodulation using MicroCT.Frontiers in cell and developmental biology · 2022Article
- Automatic and unbiased segmentation and quantification of myofibers in skeletal muscle.Scientific reports · 2021Article
- Derangements of amino acids in cachectic skeletal muscle are caused by mitochondrial dysfunction.Journal of cachexia, sarcopenia and muscle · 2020Article
- Myosoft: An automated muscle histology analysis tool using machine learning algorithm utilizing FIJI/ImageJ software.PloS one · 2020Article
- Peripheral nerve transfers change target muscle structure and function.Science advances · 2019Article
- MuscleJ: a high-content analysis method to study skeletal muscle with a new Fiji tool.Skeletal muscle · 2018Article
- The role of striated muscle Pik3r1 in glucose and protein metabolism following chronic glucocorticoid exposure.The Journal of biological chemistryArticle
Corrections and comments
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Authors and funding
7 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Quantification of muscle fiber populations provides a deeper insight into the effects of disease, trauma, and various other influences on skeletal muscle composition. Various time-consuming methods have traditionally been used to study fiber populations in many fields of research. However, recently developed immunohistochemical methods based on myosin heavy chain protein expression provide a quick alternative to identify multiple fiber types in a single section. Here, we present a rapid, reliable and reproducible protocol for improved staining quality, allowing automatic acquisition of whole cross sections and automatic quantification of fiber populations with ImageJ. For this purpose, embedded skeletal muscles are cut in cross sections, stained using myosin heavy chains antibodies with secondary fluorescent antibodies and DAPI for cell nuclei staining. Whole cross sections are then scanned automatically using a slide scanner to obtain high-resolution composite pictures of the entire specimen. Fiber population analyses are subsequently performed to quantify slow, intermediate and fast fibers using an automated macro for ImageJ. We have previously shown that this method can identify fiber populations reliably to a degree of ±4%. In addition, this method reduces inter-user variability and time per analyses significantly using the open source platform ImageJ.
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