Light sheet fluorescence microscopy (LSFM) is the best method for highspeed imaging of thick, live samples over a large field of view (FOV). In this dissertation, we describe four different projects to improve LSFM’s performance. Though LSFM provides excellent temporal resolution and optical sectioning, the images are affected by sample size and thickness. Scattering introduces background light, leading to a lower signal-to-noise ratio (SNR). We have implemented optical sectioning structured illumination (SIM), improving SNR by a factor of 5.6. Further, the system’s imaging speed can capture seizure dynamics in the central nervous system (CNS) of zebrafish larvae. Stripe artifacts in LSFM lower image quality. No current solutions are optimized for SIM-LSFM. To mitigate artifacts while improving optical sectioning capabilities for semi-opaque specimens using SIM, we developed an axial dithering approach that reduces stripe artifacts by 20% at 156 microns deep into the specimen and an adaptive reconstruction approach to improve the image with38% increase in uniformity. The volumetric imaging speed of LSFM can be accelerated through the use of an Electrical Tunable Lens (ETL) which allows the focal plane imaged onto the camera to be ] rapidly adjusted, allowing for imaging at several volumes per second. However, it also introduces spatially varying aberrations. Here, we demonstrate a system combining adaptive optics and an ETL, improving the signal to background ratio by a factor of 3.5 across a 400 by 400 by 100 μm3 volume. Further, it is fast enough to capture neural activities in the CNS of zebrafish larva. The wide-field detection nature of LSFM also limits resolution to the cellular level laterally and reduces its resolution by a factor of 3 axially. Improvement of axial resolution usually comes at the cost of FOV. We have developed a single-objective LSFM system with super-resolution SIM. This allows for multi-direction illumination with better penetration and isotropic resolution improvement. We have measured fluorescent breads and cerebellum organoids with a lateral resolution 313nm and 1.64m axial resolution over a 20μm axial range with 276μm FOV using 2D SIM. We demonstrate through simulations an achievable resolution of 161 nm by 767 nm.