title: Bioluminescent system for dynamic imaging of cell and animal behavior.
abstract: The current utility of bioluminescence imaging is constrained by a low photon yield that limits temporal sensitivity. Here, we describe an imaging method that uses a chemiluminescent/fluorescent protein, ffLuc-cp156, which consists of a yellow variant of Aequorea GFP and firefly luciferase. We report an improvement in photon yield by over three orders of magnitude over current bioluminescent systems. We imaged cellular movement at high resolution including neuronal growth cones and microglial cell protrusions. Transgenic ffLuc-cp156 mice enabled video-rate bioluminescence imaging of freely moving animals, which may provide a reliable assay for drug distribution in behaving animals for pre-clinical studies.
authors.affiliation: Department of Physiology, Keio University School of Medicine, Tokyo 160-8582, Japan.
authors.forename: Chikako
authors.initials: C
authors.lastname: Hara-Miyauchi
authors.forename: Osahiko
authors.initials: O
authors.lastname: Tsuji
authors.forename: Aki
authors.initials: A
authors.lastname: Hanyu
authors.forename: Seiji
authors.initials: S
authors.lastname: Okada
authors.forename: Akimasa
authors.initials: A
authors.lastname: Yasuda
authors.forename: Takashi
authors.initials: T
authors.lastname: Fukano
authors.forename: Chihiro
authors.initials: C
authors.lastname: Akazawa
authors.forename: Masaya
authors.initials: M
authors.lastname: Nakamura
authors.forename: Takeshi
authors.initials: T
authors.lastname: Imamura
authors.forename: Yumi
authors.initials: Y
authors.lastname: Matsuzaki
authors.forename: Hirotaka James
authors.initials: HJ
authors.lastname: Okano
authors.forename: Atsushi
authors.initials: A
authors.lastname: Miyawaki
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Okano
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis / Q000378:metabolism; D001522:Behavior, Animal; D002477:Cells / Q000737:chemistry / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000378:metabolism; D049449:Luminescence; D008164:Luminescent Proteins / Q000032:analysis / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D057054:Molecular Imaging / Q000379:methods*; D009432:Neural Crest / Q000166:cytology / Q000378:metabolism; D058953:Neural Stem Cells / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D008156:Luciferases