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Differential phase contrast microscopy (DPC) provides high-resolution quantitative phase distribution of thin transparent samples under multi-axis asymmetric illuminations. Typically, illumination in DPC microscopic systems is designed with 2-axis half-circle amplitude patterns, which, however, result in a non-isotropic phase contrast transfer function (PTF). Efforts have been made to achieve isotropic DPC by replacing the conventional half-circle illumination aperture with radially asymmetric patterns with 3-axis illumination or gradient amplitude patterns with 2-axis illumination. Nevertheless, these illumination apertures were empirically designed based on empirical criteria related to the shape of the PTF, leaving the underlying theoretical mechanisms unexplored. Furthermore, the frequency responses of the PTFs under these engineered illuminations have not been fully optimized, leading to suboptimal phase contrast and signal-to-noise ratio (SNR) for phase reconstruction. In this Letter, we provide a rigorous theoretical analysis about the necessary and sufficient conditions for DPC to achieve perfectly isotropic PTF. In addition, we derive the optimal illumination scheme to maximize the frequency response for both low and high frequencies (from 0 to 2NAobj), and meanwhile achieve perfectly isotropic PTF with only 2-axis intensity measurements. We present the derivation, implementation, simulation and experimental results demonstrating the superiority of our method over state-of-the-arts in both phase reconstruction accuracy and noise-robustness.

Optimal illumination scheme for isotropic quantitative differential phase contrast microscopy