Light Sheet Microscopy Across Scales: From Single Cells to Whole Organs

Abstract Light sheet fluorescence microscopy (LSFM), or selective plane illumination microscopy (SPIM), has revolutionized biological imaging by enabling rapid, high-resolution 3D visualization of living and fixed specimens with minimal photodamage. Since its introduction in 2004, LSFM has diversified into modalities suited for single-cell dynamics, small to medium tissues, and large, cleared specimens. This article explores how different LSFM platforms are optimized for specific sample size ranges. We compare their design features, strengths, and limitations while highlighting practical considerations for sample preparation, data handling, and system selection. Ultimately, matching LSFM modality to biological scale enhances imaging outcomes, guiding researchers in choosing the most appropriate system for their experimental needs.