Single Myofiber Isolation
Single Myofiber Isolation - Related Videos
03:53
Myofiber Isolation: A Technique to Obtain Single Myofibers from Harvested Mouse Extensor Digitorum Longus (EDL) Muscle
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2025
This video demonstrates the isolation of single myofibres from harvested murine Extensor Digitorum Longus (EDL) muscle. The isolated myofibres can be cultured to study myofiber-associated muscle stem cell activity.
07:52
DNA Electroporation, Isolation and Imaging of Myofibers
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Cited by 30 •
2015
This protocol utilizes electroporation to introduce and express fluorescently labeled proteins in mouse muscle fibers. Following recovery after electroporation, fibers are isolated. Individual fibers are then imaged using high resolution confocal microscopy to visualize muscle structure.
11:57
Isolation and Culture of Individual Myofibers and their Satellite Cells from Adult Skeletal Muscle
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Cited by 173 •
2013
Isolation and culture of myofibers is the gold standard in vitro system to study the transition of satellite cells through quiescence, activation and differentiation. Importantly, the single myofiber culture system preserves the myofiber/stem cell association, which is an essential component of the muscle stem cell niche.
08:12
In Vitro Differentiation of Mature Myofibers for Live Imaging
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Cited by 38 •
2017
Muscle cells are among the most complex eukaryotic cells. We present a protocol for the in vitro differentiation of highly mature myofibers that allows for genetic manipulation and clear imaging during all developmental stages.
05:58
Analysis of Embryonic and Larval Zebrafish Skeletal Myofibers from Dissociated Preparations
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Cited by 21 •
2013
Zebrafish are an emerging system for modeling human disorders of the skeletal muscle. We describe a fast and efficient method to isolate skeletal muscle myofibers from embryonic and larval zebrafish. This method yields a high-density myofiber preparation suitable for study of single skeletal muscle fiber morphology, protein subcellular localization, and muscle physiology.
FAQs
Collagenase loosens the extracellular matrix, the network surrounding and supporting muscle fibers in intact tissue. This enzymatic step makes the tissue less resistant to separation, allowing subsequent mechanical trituration to release individual fibers. Its role is therefore preparative: it reduces matrix constraints while helping preserve fibers for later microscopic examination or culture.
Gentle mechanical trituration is important because excessive force can compromise the intact fiber preparation, whereas controlled movement releases fibers after enzymatic loosening has reduced tissue cohesion. Preserving intact fibers matters for studying structure and function directly, including contractility and relationships with associated nuclei and surrounding cellular features.
Preserved associated nuclei and surrounding cellular features allow investigators to examine more than fiber shape alone. They can relate individual fiber structure to contractility, observe satellite cell behavior, and investigate neuromuscular interactions in a preparation centered on one fiber. This resolution helps connect cellular features with muscle function and regeneration-related responses.
A typical workflow begins with intact skeletal muscle tissue, followed by collagenase digestion to loosen the extracellular matrix. Gentle mechanical trituration then releases fibers, which are collected and examined under a microscope or maintained in culture. These stages connect tissue preparation with direct analysis of individual muscle fibers.
Microscopic examination provides a way to inspect isolated fibers and their associated cellular features directly. Culture offers a complementary setting for maintaining the fibers while studying processes such as satellite cell behavior. Choosing between these outcomes depends on whether the investigation emphasizes immediate structural observation or continued cellular analysis in the prepared sample.
The preparation is useful when researchers need to study muscle at the level of individual fibers rather than only intact tissue. In biology, it supports investigations of muscle development, aging, injury, and disease, while also enabling analysis of regeneration, neuromuscular interactions, contractility, and satellite cell behavior.