DMD-based parallel line-illumination modulation for high-throughput optical sectioning microscopy

Three-dimensional structural mapping of biological organs at single-cell resolution is essential for understanding their physiological functions and pathological mechanisms. However, conventional optical microscopy techniques face a fundamental trade-off between imaging speed and out-of-focus background suppression, making it difficult to balance high-throughput imaging of large samples with high signal-to-background ratio (SBR). To address this challenge, we present a parallel line-illumination modulation optical sectioning microscopy method (P-LiMo) based on a digital micromirror device (DMD). By employing multiple parallel modulated beams and differential computation, the method significantly improves imaging throughput while effectively suppressing defocused background signals. Compared with conventional single-beam line-illumination modulation (LiMo), P-LiMo leverages the rapid switching capability of the DMD to achieve an order-of-magnitude enhancement in imaging speed. In addition, the system systematically investigated the effects of line illumination width, spatial period, tilt angle, and reconstruction slice count on imaging quality, providing a tunable technical solution for optimizing 3D structural analysis of large specimens. This offers what we believe to be a novel tool for full-scale high-speed imaging and efficient processing of large-volume data in large-sample studies.