Protocol for multiscale in vivo adenosine triphosphate imaging in living mice

In vivo adenosine triphosphate (ATP) imaging is a powerful approach for studying physiological and pathological mechanisms underlying various metabolic processes. Here, we present a protocol for multiscale in vivo ATP imaging using AVID (ATP visualization in vivo directly) mice, which express GO-ATeam2, a fluorescent biosensor. We describe steps for endotracheal intubation, mechanical ventilation, and imaging preparation. We then detail procedures for macroscopic or two-photon imaging and image analysis.