Spatiotemporal‐Continuity‐Prior‐Guided Intensity Diffraction Tomography

Dynamic live‐cell imaging demands high spatiotemporal resolution to capture rapid intracellular processes such as organelle transport. Conventional intensity diffraction tomography (IDT) enables label‐free, quantitative 3D refractive‐index (RI) imaging but relies on sequential angular acquisitions, resulting in an inherent trade‐off between imaging speed and reconstruction fidelity. Here, we introduce spatiotemporal‐continuity‐prior‐guided IDT (SCP‐IDT), a computational framework that breaks this trade‐off by recovering intermediate 3D volumes from the same raw measurements. By leveraging the inherent spatial continuity of biological samples, SCP‐IDT implements a sliding‐window strategy to recombine adjacent illumination stacks for elevated imaging rates, while simultaneously employing Hessian‐based regularization to mitigate RI underestimation and enhance structural fidelity beyond traditional deconvolution. To further ensure seamless 4D (‐) evolution, a temporal continuity prior is imposed via a total variation constraint along the time axis, effectively bridging sequential frames. Simulations and experiments demonstrate SCP‐IDT's capability to capture 4D cellular dynamics, including mitosis and intercellular material exchange with high fidelity at camera‐limited speed, while maintaining incoherent diffraction‐limited resolution (388 nm, 40/0.75 NA). These capabilities position SCP‐IDT as a powerful tool for quantitative kinetic analysis in drug discovery and cell biology.