Abstract

Differential phase contrast (DPC) microscopy, a central quantitative phase imaging (QPI) technique in cell biology, facilitates label-free, real-time monitoring of intrinsic optical phase variations in biological samples. The existing DPC imaging theory, while important for QPI, is grounded in paraxial diffraction theory. However, this theory lacks accuracy when applied to high numerical aperture (NA) systems that are vital for high-resolution cellular studies. To tackle this limitation, we have, for the first time, formulated a nonparaxial DPC imaging equation with a transmission cross-coefficient (TCC) for high NA DPC microscopy. Our theoretical framework incorporates the apodization of the high NA objective lens, nonparaxial light propagation, and the angular distribution of source intensity or detector sensitivity. Thus, our TCC model deviates significantly from traditional paraxial TCCs, influenced by both NA and the angular variation of illumination or detection. Our nonparaxial imaging theory could enhance phase retrieval accuracy in QPI based on high NA DPC imaging.

© 2023 Optical Society of Korea

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