Single-shot polarization phase-contrast microscopy with rotational self-calibration based on multi-polarization-directional illumination

To address the fundamental limitation of traditional differential phase contrast imaging-its dependence on multi-frame acquisition, which restricts temporal resolution and hinders the observation of fast dynamic processes such as live-cell activities-we propose a single-shot Polarization Phase-Contrast microscopy with Rotational self-calibration based on multi-polarization-direction illumination. A polarization mask with three sector-shaped polarized regions enables polarization multiplexed modulation, allowing the three directional illumination components to be recovered simultaneously from a single exposure through an invertible polarization intensity transfer model, which also provides an estimate of the global rotational misalignment. The phase is then reconstructed using an alternating direction method of multipliers (ADMM) combined with total variation (TV) regularization, effectively suppressing haze-like noise and background interference commonly observed in unstained transparent samples while preserving the optical-thickness features of cellular structures, thereby improving phase reconstruction accuracy. Numerical simulations and live-cell dynamic experiments show that the proposed method substantially increases temporal resolution without degrading phase reconstruction quality, and that it maintains stable performance under complex noise conditions as well as in the presence of assembly-induced rotational errors. This approach offers a promising and practical solution for label-free quantitative phase imaging of dynamic biological processes.