VOLUMETRIC IMAGING OF CELLULAR DYNAMICS USING LIGHT-SHEET MICROSCOPY: FROM SUBCELLULAR STRUCTURES TO WHOLE ORGANISMS

Advancing biomedical imaging requires platforms that combine high-resolution, speed, and stability across biological scales. This dissertation presents three innovations designed to address limitations of conventional light-sheet and oblique plane imaging. First, we introduce Reflected Inline Detection in Epi-Oblique Plane Microscopy (RIDE-OPM), a compact and drift-resistant system that eliminates a tertiary detection path while maintaining alignment flexibility, enabling robust long-term live imaging. Second, we present a high-speed Axially Swept Light-Sheet Microscopy (ASLM) platform for cleared tissues, using dual foci and synchronized sweeping to achieve isotropic sub-micron resolution at up to 40 frames-per-second, a fourfold improvement over standard ASLM. Integrated with a deep-learning tissue boundary detector, it enables efficient 3D imaging. Finally, we propose a remote focusing architecture that ensures consistent magnification and 4f geometry across objectives, enabling seamless multi-scale imaging. Together, these methods establish a flexible framework for next-generation microscopy with broad applications in live, cleared, and multi-resolution imaging.