PySight: plug and play photon counting for fast continuous volumetric intravital microscopy

Imaging increasingly large neuronal populations at high rates pushed multi-photon microscopy into the photon-deprived regime. We present PySight, an add-on hardware and software solution tailored for photon-deprived imaging conditions. PySight more than triples the measured median amplitude of neuronal calcium transients in awake mice and facilitates single-trial intravital voltage imaging in fruit flies. Its unique data streaming architecture allowed us to image a fruit fly's brain olfactory response over 234 mu m x 600 mu m x 330 mu m at 73 volumes per second, while retaining over 200 times lower data rates than those of a conventional data acquisition system with comparable voxel sizes (1.2 mu m x 1.2 mu m x 2.2 mu m). PySight requires no electronics expertise or custom synchronization boards, and its open-source software is extensible to any imaging method based on single-pixel (bucket) detectors. PySight offers an optimal data acquisition scheme for ever increasing imaging volumes of turbid living tissue. (C) 2018 Optical Society of America under the terms of the OSA Open Access Publishing Agreement