Highly efficient quantitative phase microscopy using programmable annular LED illumination

TL;DR

This paper introduces a highly efficient quantitative phase microscopy method using programmable annular LED illumination, enhancing resolution and noise robustness for biological and biomedical imaging.

Contribution

It proposes a novel LED-based coded illumination technique that doubles resolution and improves noise robustness in quantitative phase imaging.

Findings

Achieved twice the resolution of traditional methods.

Validated accuracy with micro polystyrene beads and transmission gratings.

Demonstrated applicability on unstained human cancer cells.

Abstract

In this work, we present a highly efficient quantitative phase imaging (QPI) approach using programmable annular LED illumination based on traditional bright-field microscope. As a new type of coded illumination, the LED array provides a flexible and compatible way to realize QPI. The proposed method modulates the transfer function of system by changing the LED illumination pattern, which achieves twice resolution of objective NA and gives noise-robust response of transfer function. The phase of a sample could be recovered from the intensity images with the inversion of transfer function. Moreover, the weak object transfer function (WOTF) of axisymmetric oblique source is derived, and the noise-free and noisy simulation results sufficiently validate the applicability of discrete annular source. The quantitative phase measurements of micro polystyrene bead and visible blazed transmission grating demonstrate the accuracy of proposed method. Finally, the experimental investigations of unstained human cancer cells using different types objective are presented, and these results show the possibility of widespread adoption of QPI in the morphology study of cellular processes and biomedical community.