Deep two-photon voltage imaging with adaptive excitation

Optical imaging of neuronal voltage dynamics is invaluable to studying brain functions. However, high-speed imaging at significant depth is challenging due to the limitations of the short pixel dwell time and the maximum permissible excitation power in tissues. We report high-speed, deep voltage imaging by applying adaptive excitation, which illuminates the regions of interest only. We imaged neuronal voltage activities across two planes down to 500-630 μm depth in the awake mouse brain. Our techniques can be straightforwardly applied to typical two-photon microscopes.