Real-time imaging of moving organs and tissues at microscopic resolutions represents a major challenge in studying complex biology in live systems. Here, we present a new technique for imaging the beating murine heart at the single cell level, based on a novel stabilizer setup combined with a gating acquisition algorithm. The method allowed serial in vivo fluorescence imaging of the beating heart in live mice in both confocal and nonlinear modes for several hours. We demonstrate the utility of this technique for in vivo optical sectioning and dual-channel time-lapse fluorescence imaging of cardiac ischemia. The generic method could be adapted to other moving organs and thus broadly facilitate in vivo microscopic investigations.

Highlights d Ultrasound-induced neuromodulation is initiated by opening of TRPA1 in astrocytes d The Ca 2+ entry through TRPA1 causes a release of glutamate through Best1 channels d The released glutamate activates NMDA receptors in neighboring neurons