Time-lapse imaging using dual-color coded quantitative differential phase contrast microscopy
Journal
Proceedings of SPIE - The International Society for Optical Engineering
Journal Volume
12151
ISBN
9781510651784
Date Issued
2022-01-01
Author(s)
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.
Subjects
biomedical imaging | phase contrast imaging | pupil design | quantitative
Type
conference paper