Single‐shot quantitative phase imaging as an extension of differential interference contrast microscopy

Abstract

Various studies have been conducted to obtain quantitative phase information based on differential interference contrast (DIC) microscopy. As one such attempt, we propose in this study a single‐shot quantitative phase imaging (QPI) method by combining two developments. First, an add‐on optical system to a commercialized DIC microscope was developed to perform quantitative phase gradient imaging (QPGI) with single image acquisition using a polarization camera. Second, an algorithm was formulated to reconstitute QPI from the obtained QPGI by reducing linear artifacts, which arise in simply integrated QPGI images. To demonstrate the applicability of the developed system in cell biology, the system was used to measure various cell lines and compared with fluorescence microscopy images of the same field of view. Consistent with previous studies, nucleoli and lipid droplets can be imaged by the system with greater optical path lengths (OPL). The results also implied that combining fluorescence microscopy and the developed system might be more informative for cell biology research than using these methods individually. Exploiting the single‐shot performance of the developed system, time‐lapse imaging was also conducted to visualize the dynamics of intracellular granules in monocyte‐/macrophage‐like cells. Our proposed approach may accelerate the implementation of QPI in standard biomedical laboratories.

Capturing cell morphology dynamics with high temporal resolution using single-shot quantitative phase gradient imaging

“…The optical system for ss-QPGM was built as a functional extension of a DIC microscope, 11 , 12 with the configuration shown in Fig. 1(a) .…”

“…An alternative approach that achieved single-shot QPI utilizing a quarter-wave plate and a polarization camera has been demonstrated. 12 Since the integration of quantitative phase gradients can induce image artifacts, the authors employed the Alternative Direction Method of Multipliers-based algorithm for phase recovery in this work; however, this processing requires extensive computing resources and time. Efforts to achieve real-time QPI images using light-emitting diode (LED)-based color-multiplexing, 13 , 14 polarization multiplexing, 15 or meta-surfaces 16 have also been reported.…”

Quantitative phase imaging by gradient retardance optical microscopy

“…More recently, DIC-based QPI was combined with phase shifting interferometry (PSI) to enhance sensitivity and robustness. In this context, PSI was enabled by modifying the imaging path using combinations of quarter waveplates and a rotatable analyzer 38 or polarization sensitive cameras 39 – 42 , as well as spatial light modulators in the imaging path 23 . Alternative approaches include specimen rotation 43 – 45 or modifying both the illumination and detection paths with a liquid crystal polarization rotator between the Wollaston prisms and polarizers 46 .…”

Non-invasive Visualization and Characterization of Bile Canaliculus Formation Using Refractive Index Tomography

“…15) RI serves as a cellular intrinsic imaging contrast that reflects morphological and biophysical information. 15,16) RI tomography can acquire the 3D distribution of RI with high resolution, and visualize subcellular structures, including chromosomes, mitochondria, and lipid droplets, in living cells. 17,18) Although the observation target of RI tomography was basically limited to the single cell level, we recently developed an in-silico clearing approach for deep RI tomography, which suppresses multiple-scattering via corrective light propagation in a computer.…”