Hybrid light-sheet and light-field microscope for high resolution and large volume neuroimaging

Large scale simultaneous recording of fast patterns of neural activity remains challenging. Volumetric imaging modalities such as scanning-beam light-sheet microscopy (LSM) and wide-field light-field microscopy (WFLFM) fall short of the goal due to their complex calibration procedure, low spatial resolution, or high-photobleaching. Here, we demonstrate a hybrid light-sheet light-field microscopy (LSLFM) modality that yields high spatial resolution with simplified alignment of the imaging plane and the excitation plane. This new modality combines the selective excitation of light-sheet illumination with volumetric light-field imaging. This modality overcomes the current limitations of the scanning-beam LSM and WFLFM implementations. Compared with LSM, LSLFM captures volumetric data at a frame rate 50× lower than the rate of LSM and requires no dynamic calibration. Compared with WFLFM, LSLFM produces moderate improvements in spatial resolutions, 10 times improvement in the contrast when imaging fluorescent beads, and 3.2× the signal-to-noise ratio in the detection of neural activity when imaging live zebrafish expressing a genetically encoded calcium sensor.