Using Electrically Tunable Lens to Improve Axial Resolution and Imaging Field in Light Sheet Fluorescence Microscope

Light sheet fluorescence microscope (LSFM) has become one of the most promising 3D microscopy techniques due to its prominent lateral section illumination. In addition, comparing with the confocal microscope, it has low phototoxicity and high temporal resolution. In this paper, we proposed an easy-to-operate tiling light sheet fluorescence microscope, which integrates a cylindrical lens, a galvanometric mirror, and an excitation objective to produce a light sheet with desired numerical aperture and axial resolution and utilizes an electrically tunable lens (ETL) to enlarge the limited field of view (FOV) of single excitation section following the temporal multiplexing. Imaging performances have been characterized from the experiment by imaging the 20 nm fluorescent beads with water immersion, and the benefits of our proposed technique have been shown in volumetric resolution, FOV, and speed as well as contrast.