Single-shot color-coded LED microscopy for quantitative differential phase contrast imaging

Common quantitative differential phase contrast microscopy imaging usually uses multistep circular or annular illuminations and color multiplexed illuminations, bringing about low capture efficiency and poor phase reconstruction, respectively. We present a new single-shot quantitative differential phase contrast microscopy imaging method, which uses R/B-G annular LED multiplexed illumination to achieve better phase reconstruction by capturing 1 colored image. By deriving the phase transfer function of the illumination and simulation verification, the superiority of it is confirmed. Through phase reconstruction simulations for circular step and gradual samples, we verify that the R/B-G annular multiplexed illumination has better axial phase reconstruction accuracy and weaker lateral anisotropy than other color multiplexed illuminations. As a demonstration, quantitative phase imaging is performed on Quantitative Phase Target, validating the axial accuracy, lateral resolution, and lateral anisotropy of our method. Imaging and phase reconstruction were also performed on mouse kidney tissue slice and motional silica microspheres to visually express the utility and practicability of this method. Our research provides a better color multiplexed avenue for the real-time quantitative phase imaging in dispersionfree, weak-phase samples, especially living biological tissues.

Automated summary

Common quantitative differential phase contrast microscopy imaging usually suffers from low capture efficiency and poor phase reconstruction due to multistep circular or annular illuminations and color multiplexed illuminations. In this study, a new single-shot imaging method using R/B-G annular LED multiplexed illumination is presented for better phase reconstruction. Simulation results show that this method achieves better axial phase reconstruction accuracy and weaker lateral anisotropy compared to other color multiplexed illuminations. Experimental results on different samples demonstrate the feasibility and utility of this method in real-time quantitative phase imaging, especially for living biological tissues.