title: Application of FRET-Based Biosensor "ATeam" for Visualization of ATP Levels in the Mitochondrial Matrix of Living Mammalian Cells.
abstract: Genetically encoded biosensors utilizing the Förster resonance energy transfer (FRET) are powerful tools for live cell imaging of various cellular processes. Our group has previously developed a series of FRET-based biosensors, named "ATeam," for visualization of ATP levels inside a single living cell. ATeam not only provides a window of insight into a single cell but also allows for visualization of ATP levels in mitochondrial matrix of a single living cell. This novel tool is able to monitor alterations in cellular ATP in response to various treatments in real time. Here we present a method for the evaluation of ATP levels in mitochondria in living cells by using ATeam. At the end of this chapter, an example of experimental results is described for a better understanding of the presented procedure.
authors.affiliation: Department of Functional Biology, Graduate School of Biostudies, Kyoto University, Yoshida-konoe-cho, Sakyo-ku, Kyoto, 606-8501, Japan.
authors.forename: Tomoki
authors.initials: T
authors.lastname: Yoshida
authors.affiliation: Department of Functional Biology, Graduate School of Biostudies, Kyoto University, Yoshida-konoe-cho, Sakyo-ku, Kyoto, 606-8501, Japan.
authors.forename: Soaad
authors.initials: S
authors.lastname: Alfaqaan
authors.affiliation: Department of Functional Biology, Graduate School of Biostudies, Kyoto University, Yoshida-konoe-cho, Sakyo-ku, Kyoto, 606-8501, Japan.
authors.forename: Norio
authors.initials: N
authors.lastname: Sasaoka
authors.affiliation: Department of Functional Biology, Graduate School of Biostudies, Kyoto University, Yoshida-konoe-cho, Sakyo-ku, Kyoto, 606-8501, Japan. imamura@lif.kyoto-u.ac.jp.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Imamura
mesh_terms: D000255:Adenosine Triphosphate / Q000032:analysis*; D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002478:Cells, Cultured; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism*; D057054:Molecular Imaging / Q000295:instrumentation / Q000379:methods; D013223:Statistics as Topic
chemical_list: D000255:Adenosine Triphosphate
keywords: ATP; ATeam; FRET; Fluorescent biosensor; Live imaging