Here, we present a rapid dual-view projection imaging method with two-photon glutamate uncaging capability based on an upright Bessel beam light-sheet microscopy setup. Compared with light-sheet imaging, our projection imaging method can significantly improve the volumetric rate for neural imaging. In addition, an independent laser scanning module is designed for 2P uncaging, allowing simultaneous synaptic resolution of stimulation and 100 Hz volumetric imaging of neural activity in deep tissue. Imaging results from mouse brain slices under 2P glutamate uncaging will be presented.
Light-sheet fluorescence microscopy serves as a fast high-resolution imaging method for neural imaging. However, its 3D imaging ability is often limited by the speed of scanning the detection focal plane in the z-direction. Herein, we develop a rapid random z-access two-photon light-sheet microscope, which incorporates a two-photon Bessel beam light-sheet microscope with a dynamically driven electrical tunable lens (ETL). With a precise ETL calibration process and a novel rapid random z-access method, our system can selectively scan any desired z-section in the 3D imaging volume at the speed of 100 frame-per-second, and allows neural activates monitoring on the living brain tissue at video rate.
Neuroscience research is propelled by advance imaging tools, which ideally could provide high spatial and temporal resolution imaging. However, available fast 3-dimensional optical sectioning approaches applicable to deep tissue and large scale in-vivo imaging are still limited. We develop a novel two-photon Bessel beam scanning light sheet microscope, which has two distinct imaging modes: 1) The light-sheet imaging mode enables sub-micron 3D imaging over hundreds of microns field of view, which is well adapted to the neural tissue with complex structure; 2) The projection imaging mode aims to capture fast neural activities at up to 100 volumes per second. Imaging results of various neural specimens will be presented.
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