Application of FRET-Based Biosensor “ATeam” for Visualization of ATP Levels in the Mitochondrial Matrix of Living Mammalian Cells

• DOI: 10.1007/978-1-4939-6824-4_14
• Publication Date: 2017-03-08T11:51:57Z
• Authors: Tomoki Yoshida; Soaad Alfaqaan; Norio Sasaoka; Hiromi Imamura
• Keywords: Adenosine Triphosphate; Microscopy, Fluorescence; Statistics as Topic; Fluorescence Resonance Energy Transfer; Image Processing, Computer-Assisted; Animals; Humans; Biosensing Techniques; Cells, Cultured; Mitochondria; Molecular Imaging

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.