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A miniaturized version of a light-sheet microscopy (LSM) system, with 3D imaging enabled through active optical control, is presented. Even though the field of LSM technology has advanced significantly in recent years, it is still not considered an easily available technique. This is mainly due to its cost compared to epifluorescence setups and the requirement for specific sample mounting techniques in most cases, as well as stringent optical alignment and difficulty to reduce motion artifacts when the sample is moved through the light path to create the imaging slices. In our research, we demonstrate a miniaturized version of an LSM that can reduce size and cost, and is able to achieve 3D imaging through control of multiple active optical elements and MEMS micromirrors used in both the illumination and imaging path instead of moving the sample. The laser excitation is controlled and shaped via multiple MEMS elements for 3D beam position control and multi-lens beam shaping to generate a 2.85 μm wide light-sheet with controllable height of up to 550 μm, and orthogonal positioning over a 200 μm range. Additionally, the focal point of the excitation can be shifted along the laser propagation direction by 200 μm. The orthogonally positioned imaging path incorporates a x20, NA = 0.4 objective and a tunable lens for imaging selected focal planes synchronized with the excitation positioning. The imaging results show sub-micron resolution with a field-of-view of 400 μm x 300 μm. The synchronization of the two active elements allows for fast imaging of different slices of a sample and promises convenient 3D reconstruction and representation of cell tissue.

Light-sheet microscopy using MEMS and active optics for 3D image acquisition control