Fig. 1. Development of light sheet microscopy in high-throughput imaging. (a) Adult GFP-M mouse was transcardially perfused, cleared with uDISCO and imaged by fluorescence stereomicroscope at cellular resolution^[5]; (b) working principle and comparison of axial image quality achieved in a CLARITY-cleared VIP-tdTomato mouse brain for standard light sheet illumination (left) and for the axially swept light sheet mode (right) ^[7]; (c) working principle of the proposed method, tiling a smaller but thinner light sheet along the light sheet length direction and extending the field of view (lateral and axial maximum intensity projections of a cleared Thy1-YFP mouse brain imaged with a Zeiss light sheet microscope, a six-tile mode, and a non-tile mode of the presented tiling light sheet microscope)^[10]; (d) CACS procedure and the comparison of CACS Bessel plane illumination microscopy and other imaging modes^[18]

Fig. 2. Application of light sheet microscope in super-resolution imaging. (a) Simulation results of the intensity pattern at the rear pupil plane of the excitation objective, the cross-sectional intensity of the pattern in the xz plane at the focus of the excitation objective, the cross-sectional intensity of the light sheet created by dithering the focal pattern along the x axis, and the xz cross section of the overall PSF of the microscope of the traditional Gaussian light sheet, Bessel swept sheet, the square lattice and the hexagonal lattice^[25]; (b) conventional SPIM image and IML-SPIM image of human mammary MCF10A cell spheroids expressing H2B-PAmCherry^[27]; (c) principle of two-photon light sheet nanoscopy and setup of 2P3A-DSLM, the principle of TAG-based two photon light sheet excitation, generated by rapid scanning of laser focus along the optics axis (x axis) using a TAG and vertical scanning along the y-axis by a galvanometer, fast 3D imaging is achieved with a P2T^[30]; (d) schematic illustrating the generation of DRSPIM and its difference from a lattice light sheet and field synthesis. The yz slices of the microtubules (labeled with Tubulin-Atto 488) in a live U2OS cell imaged via N-SIM under the wide-field mode and 3D N-SIM mode DR-SPIM and SRRF DR–SPIM^[32]

Fig. 3. Typical modality of light sheet microscope. (a) Inverted T-shaped mode^[1]; (b) V-shaped double objective; (c) open-top double objective; (d) single objective light sheet microscope^[33]