DNA Electroporation, Isolation and Imaging of Myofibers

Overview

Journal J Vis Exp

Specialties Biology
General Medicine

Date 2016 Jan 19

PMID 26780499

Citations 21

Authors

Alexis R Demonbreun

Elizabeth M McNally

Affiliations

Soon will be listed here.

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.

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