Imaging the peripheral and central nervous system requires high-resolution techniques capable of resolving fine axonal structures and synaptic connections across large, three-dimensional volumes. These tissues pose unique challenges due to their dense, heterogeneous composition and the intricate, branching architecture of nerve fibers, necessitating imaging approaches that combine deep tissue penetration, isotropic resolution, and minimal photodamage. This chapter outlines a robust protocol for tissue clearing and imaging using axially swept light-sheet microscopy (ASLM). We describe the optical principles of light-sheet fluorescence microscopy (LSFM), the challenges of imaging cleared tissues, and the critical trade-offs between field of view, resolution, and optical sectioning. By comparing the performance of two ASLM systems—bench-top mesoSPIM and a custom-designed cleared tissue (CT)-ASLM—we evaluate their capabilities in imaging complex neural and organ systems. Quantitative analyses of image sharpness, staining uniformity, and aggregate reduction serve as valuable metrics for optimizing tissue labeling, clearing, and microscope operation. This chapter provides practical guidelines for implementing LSFM in biological research, offering insights into optimizing tissue clearing protocols, imaging parameters, and downstream data analysis to advance the study of cellular and subcellular structures within native tissue contexts.