DNA electroporation, isolation and imaging of myofibers

Alexis R. Demonbreun*, Elizabeth M. McNally

*Corresponding author for this work

Research output: Contribution to journalArticle

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 languageEnglish (US)
Article numbere53551
JournalJournal of Visualized Experiments
Volume2015
Issue number106
DOIs
StatePublished - 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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