title: Photoconvertible fluorescent protein-based live imaging of mitochondrial fusion.
abstract: Mitochondria are highly dynamic organelles that undergo fusion and fission on a relatively fast time scale. Here, a straightforward method is described for capturing mitochondrial fusion events in real time using a photoconvertible fluorescent protein and a far-field fluorescence microscope equipped with appropriate image acquisition and analysis software. The Kaede photoconvertible fluorescent protein is tagged with a mitochondrial targeting sequence and delivered to primary neurons by lentiviral transduction, which ensures efficient low copy number transgene insertion, as well as stable transgene expression.
authors.affiliation: Department of Neurosurgery, David Geffen School of Medicine at the University of California, Los Angeles, 300 Stein Plaza, Ste. 562, Los Angeles, CA, 90095, USA, geoffreyowens@mednet.ucla.edu.
authors.forename: Geoffrey C
authors.initials: GC
authors.lastname: Owens
authors.forename: David B
authors.initials: DB
authors.lastname: Edelman
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D005822:Genetic Vectors / Q000235:genetics; D007091:Image Processing, Computer-Assisted; D008164:Luminescent Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D063154:Mitochondrial Dynamics; D057054:Molecular Imaging / Q000379:methods; D009457:Neuroglia / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D051381:Rats; D014161:Transduction, Genetic
chemical_list: D008164:Luminescent Proteins