DNA electroporation, isolation and imaging of myofibers

Alexis R. Demonbreun; Elizabeth M. McNally

doi:10.3791/53551

DNA electroporation, isolation and imaging of myofibers

Alexis R. Demonbreun^*, Elizabeth M. McNally

^*Corresponding author for this work

Medicine, Cardiology Division

Research output: Contribution to journal › Article

8 Scopus citations

Abstract

Mature muscle has a unique structure that is amenable to live cell imaging. Herein, we describe the experimental protocol for expressing fluorescently labeled proteins in the flexor digitorum brevis (FDB) muscle. Conditions have been optimized to provide a large number of high quality myofibers expressing the electroporated plasmid while minimizing muscle damage. The method employs fluorescent tags on various proteins. Combining this expression method with high resolution confocal microscopy permits live cell imaging, including imaging after laser-induced damage. Fluorescent dyes combined with imaging of fluorescently-tagged proteins provides information regarding the basic structure of muscle and its response to stimuli.

Original language	English (US)
Article number	e53551
Journal	Journal of Visualized Experiments
Volume	2015
Issue number	106
DOIs	https://doi.org/10.3791/53551
State	Published - Dec 23 2015

Keywords

Confocal microscopy
DNA electroporation
Fluorescence
Issue 106
Molecular Biology
Muscle
Myofiber
Sarcolemma

ASJC Scopus subject areas

Neuroscience(all)
Chemical Engineering(all)
Biochemistry, Genetics and Molecular Biology(all)
Immunology and Microbiology(all)

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Demonbreun, A. R., & McNally, E. M. (2015). DNA electroporation, isolation and imaging of myofibers. Journal of Visualized Experiments, 2015(106), [e53551]. https://doi.org/10.3791/53551

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