Time-lapse imaging using dual-color coded quantitative differential phase contrast microscopy

Ying-Ju Chen; Yu-Zi Lin; Sunil Vyas; Tai-Horng Young; Yuan Luo

doi:10.1117/12.2624352

24 May 2022 Time-lapse imaging using dual-color coded quantitative differential phase contrast microscopy

Ying-Ju Chen, Yu-Zi Lin, Sunil Vyas, Tai-Horng Young, Yuan Luo

Proceedings Volume 12151, Photosensitive Materials and their Applications II; 1215108 (2022) https://doi.org/10.1117/12.2624352
Event: SPIE Photonics Europe, 2022, Strasbourg, France

Abstract

Quantitative differential phase contrast (qDPC) microscopy is utilized to observe label-free specimens by asymmetric illumination and intensity measurements. To provide uniform phase contrast images with minimum acquisition time, dual-color linear gradient pupils are applied to generate structured light. It is shown that the proposed pupils in qDPC can outperform half-circle and vortex pupils, and isotropic phase transfer function can be achieved with only 2-axis measurements. With the implantation of digital pupils in our system, the limitation of the time-consuming multi-axis measurement and reconstruction artifacts caused by missing frequencies in a half-circle pupil can be overcome. The required frame of dual-color coded microscopy reduces to two due to the color encoding method used in pupil design so that one-axis information can be obtained within a single shot. The improvement in imaging speed can help in monitoring fast developmental cell processes for various biological applications. Standard micro-lens-array and rat astrocyte cells were used to evaluate the performance of our microscopy system. We demonstrated time-lapse phase imaging of living cells and observed detailed morphology and dynamics changes. Present studies show the potential of the dual-color coded qDPC system for quantitative biomedical imaging for cell research. Due to our label-free approach, the natural contents of the cells remain intact and the dynamic processes through the samples can readily be observed. With our method, the nature of the structural changes in the samples can be evaluated in terms of quantitative phase changes inside acquired images.

Citation Download Citation

Ying-Ju Chen, Yu-Zi Lin, Sunil Vyas, Tai-Horng Young, and Yuan Luo "Time-lapse imaging using dual-color coded quantitative differential phase contrast microscopy", Proc. SPIE 12151, Photosensitive Materials and their Applications II, 1215108 (24 May 2022); https://doi.org/10.1117/12.2624352

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KEYWORDS

Microscopy

Phase contrast

Image processing

Microscopes

Time lapse microscopy

Imaging systems

Light sources

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