Photoconvertible fluorescent protein-based live imaging of mitochondrial fusion
Geoffrey C Owens1, David B Edelman
1Department 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.
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.
