Accelerated dynamic light sheet microscopy: unifying time-varying patterned illumination and low-rank and sparsity constrained reconstruction

Abstract Light Sheet Fluorescence Microscopy (LSFM) enables rapid and gentle 3D fluorescence imaging of dynamic processes over extended periods in translucent samples at the mesoscopic scale. However, its temporal resolution is constrained by the sequential acquisition of individual two-dimensional planes at varying depths, making it challenging to capture rapid dynamics such as the beating of a zebrafish heart. To address this limitation, we recently developed spatially modulated Selective Volume Illumination Microscopy, which utilizes a compressed sensing approach to reconstruct the entire imaging volume from measurements where multiple planes are illuminated simultaneously using spatially modulated light. Building on this advancement, we now introduce a novel spatio-temporal patterned illumination strategy and volume reconstruction method that incorporates low-rank and sparsity constraints, effectively leveraging the temporal and spatial redundancy present in sequential volumetric acquisitions. This method was applied to the volumetric imaging of embryonic zebrafish hearts, achieving an improvement in imaging speed of 4-fold compared to standard LSFM and a 2-fold improvement compared to traditional compressed sensing approaches, while preserving reconstruction accuracy and enabling the visualization of fast dynamic events with a resolution of a few tens of milliseconds. Our approach represents a step forward in enhancing the temporal resolution ofLSFM for studying fast biological dynamics.