title: DNA Electroporation, Isolation and Imaging of Myofibers.
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.
authors.affiliation: Center for Genetic Medicine, Northwestern University; alexis.demonbreun@northwestern.edu.
authors.forename: Alexis R
authors.initials: AR
authors.lastname: Demonbreun
authors.affiliation: Center for Genetic Medicine, Northwestern University.
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: McNally
mesh_terms: D000818:Animals; D004247:DNA / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D018274:Electroporation / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D018485:Muscle Fibers, Skeletal / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D018482:Muscle, Skeletal / Q000166:cytology; D010957:Plasmids / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics
chemical_list: D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D004247:DNA