KyeoReh Lee

Quantitative Phase Imaging Techniques for the Study of Cell Pathophysiology: From Principles to Applications

A cellular-level study of the pathophysiology is crucial for understanding the mechanisms behind human diseases. Recent advances in quantitative phase imaging (QPI) techniques show promises for the cellular-level understanding of the pathophysiology of diseases. To provide important insight on how the QPI techniques potentially improve the study of cell pathophysiology, here we present the principles of QPI and highlight some of the recent applications of QPI ranging from cell homeostasis to infectious diseases and cancer.

Comparative study of iterative reconstruction algorithms for missing cone problems in optical diffraction tomography

In optical tomography, there exist certain spatial frequency components that cannot be measured due to the limited projection angles imposed by the numerical aperture of objective lenses. This limitation, often called as the missing cone problem, causes the under-estimation of refractive index (RI) values in tomograms and results in severe elongations of RI distributions along the optical axis. To address this missing cone problem, several iterative reconstruction algorithms have been introduced exploiting prior knowledge such as positivity in RI differences or edges of samples. In this paper, various existing iterative reconstruction algorithms are systematically compared for mitigating the missing cone problem in optical diffraction tomography. In particular, three representative regularization schemes, edge preserving, total variation regularization, and the Gerchberg-Papoulis algorithm, were numerically and experimentally evaluated using spherical beads as well as real biological samples; human red blood cells and hepatocyte cells. Our work will provide important guidelines for choosing the appropriate regularization in ODT.

Diffraction optical tomography using a quantitative phase imaging unit

A simple and practical method to measure three-dimensional (3-D) refractive index (RI) distributions of biological cells is presented. A common-path self-reference interferometry consisting of a compact set of polarizers is attached to a conventional inverted microscope equipped with a beam scanning unit, which can precisely measure multiple 2-D holograms of a sample with high phase stability for various illumination angles, from which accurate 3-D optical diffraction tomograms of the sample can be reconstructed. 3-D RI tomograms of nonbiological samples such as polystyrene microspheres, as well as biological samples including human red blood cells and breast cancer cells, are presented.

Synthetic Fourier transform light scattering.

We present synthetic Fourier transform light scattering, a method for measuring extended angle-resolved light scattering (ARLS) from individual microscopic samples. By measuring the light fields scattered from the sample plane and numerically synthesizing them in Fourier space, the angle range of the ARLS patterns is extended up to twice the numerical aperture of the imaging system with unprecedented sensitivity and precision. Extended ARLS patterns of individual microscopic polystyrene beads, healthy human red blood cells (RBCs), and Plasmodium falciparum-parasitized RBCs are presented.

Measuring optical transmission matrices by wavefront shaping

We introduce a simple but practical method to measure the optical transmission matrix (TM) of complex media. The optical TM of a complex medium is obtained by modulating the wavefront of a beam impinging on the complex medium and imaging the transmitted full-field speckle intensity patterns. Using the retrieved TM, we demonstrate the generation and linear combination of multiple foci on demand through the complex medium. This method will be used as a versatile tool for coherence control of waves through turbid media.

High-Resolution 3-D Refractive Index Tomography and 2-D Synthetic Aperture Imaging of Live Phytoplankton

Optical measurements of the morphological and biochemical imaging of phytoplankton are presented. Employing quantitative phase imaging techniques, 3-D refractive index maps and high-resolution 2-D quantitative phase images of individual live phytoplankton are simultaneously obtained without exogenous labeling agents. In addition, biochemical information of individual phytoplankton including volume, mass, and density of individual phytoplankton are also quantitatively obtained from the measured refractive index distributions. We expect the present method to become a powerful tool for the study of phytoplankton.

Time-multiplexed structured illumination using a DMD for optical diffraction tomography

We present a time-multiplexing structured illumination control technique for optical diffraction tomography (ODT). Instead of tilting the angle of illumination, time-multiplexed sinusoidal illumination is exploited using a digital micromirror device (DMD). The present method effectively eliminates unwanted diffracted beams from binary DMD patterns, which deteriorates the image quality of the ODT in the previous binary Lee hologram method. We experimentally show the feasibility and advantage of the present method by reconstructing three-dimensional refractive index distributions of various samples and comparing with a conventional Lee hologram method.

Optogenetic control of cell signaling pathway through scattering skull using wavefront shaping

We introduce a non-invasive approach for optogenetic regulation in biological cells through highly scattering skull tissue using wavefront shaping. The wavefront of the incident light was systematically controlled using a spatial light modulator in order to overcome multiple light-scattering in a mouse skull layer and to focus light on the target cells. We demonstrate that illumination with shaped waves enables spatiotemporal regulation of intracellular Ca2+ level at the individual-cell level.

Common-path diffraction optical tomography with a low-coherence illumination for reducing speckle noise

Common-path diffraction optical tomography (cDOT) is a non-invasive and label-free optical holographic technique for measuring both the three-dimensional refractive index (RI) tomograms and two-dimensional dynamic phase images of a sample. Due to common-path geometry, cDOT provides quantitative phase imaging with high phase sensitivity. However, the image quality of the cDOT suffers from speckle noise; the use of a monochromatic laser inevitably results in the formation of parasitic fringe patterns in measured quantitative phase images. Here, we present a technique to reduce speckle noise in the cDOT using a low-coherence illumination source. Utilizing a Ti-sapphire pulsed laser in the cDOT, we achieved the reduction of speckle noise in both the three-dimensional RI tomograms and two-dimensional dynamic phase images.

In vivo deep tissue imaging using wavefront shaping optical coherence tomography

Abstract. Multiple light scattering in tissue limits the penetration of optical coherence tomography (OCT) imaging. Here, we present in vivo OCT imaging of a live mouse using wavefront shaping (WS) to enhance the penetration depth. A digital micromirror device was used in a spectral-domain OCT system for complex WS of an incident beam which resulted in the optimal delivery of light energy into deep tissue. Ex vivo imaging of chicken breasts and mouse ear tissues showed enhancements in the strength of the image signals and the penetration depth, and in vivo imaging of the tail of a live mouse provided a multilayered structure inside the tissue.