DNA Electroporation, Isolation and Imaging of Myofibers | Zendy

Alexis R. Demonbreun | Zendy; Elizabeth M. McNally | Zendy

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DNA Electroporation, Isolation and Imaging of Myofibers

Author(s) -

Alexis R. Demonbreun,

Elizabeth M. McNally

Publication year - 2015

Publication title -

journal of visualized experiments

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.596

H-Index - 91

ISSN - 1940-087X

DOI - 10.3791/53551

Subject(s) - electroporation , confocal , microbiology and biotechnology , live cell imaging , confocal microscopy , fluorescence lifetime imaging microscopy , biology , fluorescent protein , cell , fluorescence microscope , fluorescence , biophysics , green fluorescent protein , biochemistry , gene , physics , geometry , mathematics , quantum mechanics

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.

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