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Kyoohyun Kim

Publications and source records attributed to Kyoohyun Kim.

At least 19 recordsLinked to original sources

Super-resolution three-dimensional fluorescence and optical diffraction tomography of live cells using structured illumination generated by a digital micromirror device

We present a multimodal approach for measuring the three-dimensional (3D) refractive index (RI) and fluorescence distributions of live cells by combining optical diffraction tomography (ODT) and 3D structured illumination microscopy (SIM). A digital micromirror device is utilized to generate structured illumination patterns for both ODT and SIM, which enables fast and stable measurements. To verify its feasibility and applicability, the proposed method is used to measure the 3D RI distribution and 3D fluorescence image of various samples, including a cluster of fluorescent beads, and the time-lapse 3D RI dynamics of fluorescent beads inside a HeLa cell, from which the trajectory of the beads in the HeLa cell is analyzed using spatiotemporal correlations.

physics.optics

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

We present a novel illumination control technique for optical diffraction tomography (ODT). Various spatial frequencies of beam illumination were controlled by displaying time-averaged sinusoidal patterns using a digital micromirror device (DMD). Compared to the previous method using binary Lee holograms, the present method eliminates unwanted diffracted beams which may deteriorate the image quality of the ODT. We demonstrated the capability of the present method by reconstructing three-dimensional refractive index (RI) distributions of various samples, with high RI sensitivity (σ_Δn = 3.15 +/- 10-4), and reconstructing 3-D RI tomograms of biological samples, which provided quantitative biochemical and morphological information about the samples.

physics.bio-ph

Three-dimensional label-free imaging and quantification of lipid droplets in live hepatocytes

Lipid droplets (LDs) are subcellular organelles with important roles in lipid storage and metabolism and involved in various diseases including cancer, obesity, and diabetes. Conventional methods, however, have limited ability to provide quantitative information on individual LDs and have limited capability for three-dimensional (3-D) imaging of LDs in live cells especially for fast acquisition of 3-D dynamics. Here, we present an optical method based on 3-D quantitative phase imaging to measure the 3-D structural distribution and biochemical parameters (concentration and dry mass) of individual LDs in live cells without using exogenous labelling agents. The biochemical change of LDs under oleic acid treatment was quantitatively investigated, and 4-D tracking of the fast dynamics of LDs revealed the intracellular transport of LDs in live cells.

physics.bio-ph

Tomographic active optical trapping of arbitrarily shaped objects by exploiting 3-D refractive index maps

Optical trapping can be used to manipulate the three-dimensional (3-D) motion of spherical particles based on the simple prediction of optical forces and the responding motion of samples. However, controlling the 3-D behaviour of non-spherical particles with arbitrary orientations is extremely challenging, due to experimental difficulties and the extensive computations. Here, we achieved the real-time optical control of arbitrarily shaped particles by combining the wavefront shaping of a trapping beam and measurements of the 3-D refractive index (RI) distribution of samples. Engineering the 3-D light field distribution of a trapping beam based on the measured 3-D RI map of samples generates a light mould, which can be used to manipulate colloidal and biological samples which have arbitrary orientations and/or shapes. The present method provides stable control of the orientation and assembly of arbitrarily shaped particles without knowing a priori information about the sample geometry. The proposed method can be directly applied in biophotonics and soft matter physics.

physics.optics

The effects of spatiotemporal coherence on interferometric imaging

Illumination coherence plays a major role in various imaging systems, from microscopy, metrology, digital holography, optical coherence tomography, to ultrasound imaging. Here, we present a systematic study on the effects of degrees of spatiotemporal coherence of an illumination (DSTCI) on imaging quality. An optical field with arbitrary DSTCI was decomposed into wavelets with constituent spatiotemporal frequencies, and the effects on image quality were quantitatively investigated. The results show the synergistic effects on reduction of speckle noise when DSTCI is decreased. This study presents a method to systematically control DSTCI, and the result provides an essential reference on the effects of DSTCI on imaging quality. We believe that the presented methods and results can be implemented in various imaging systems for characterising and improving imaging quality.

physics.optics

Holographic intravital microscopy for 2-D and 3-D imaging intact circulating blood cells in microcapillaries of live mice

Intravital microscopy is an essential tool that reveals behaviours of live cells under conditions close to natural physiological states. So far, although various approaches for imaging cells in vivo have been proposed, most require the use of labelling and also provide only qualitative imaging information. Holographic imaging approach based on measuring the refractive index distributions of cells, however, circumvent these problems and offer quantitative and label-free imaging capability. Here, we demonstrate in vivo two- and three-dimensional holographic imaging of circulating blood cells in intact microcapillaries of live mice. The measured refractive index distributions of blood cells provide morphological and biochemical properties including three-dimensional cell shape, haemoglobin concentration, and haemoglobin contents at the individual cell level. With the present method, alterations in blood flow dynamics in live healthy and sepsis-model mouse were also investigated.

physics.bio-ph

Optical characterization of red blood cells from individuals with sickle cell trait and disease in Tanzania using quantitative phase imaging

Sickle cell disease (SCD) is common across Sub-Saharan Africa. However, the investigation of SCD in this area has been significantly limited mainly due to the lack of research facilities and skilled personnel. Here, we present optical measurements of individual red blood cells (RBCs) from healthy individuals and individuals with SCD and sickle cell trait in Tanzania using the quantitative phase imaging technique. By employing a quantitative phase imaging unit (QPIU), an existing microscope in a clinic is transformed into a powerful quantitative phase microscope providing measurements on the morphological, biochemical, and biomechanical properties of individual cells. The present approach will open up new opportunities for cost-effective investigation and diagnosis of several diseases in low resource environments.

physics.med-ph

White-light Quantitative Phase Imaging Unit

We introduce the white light quantitative phase imaging unit (WQPIU) as a practical realization of quantitative phase imaging (QPI) on standard microscope platforms. The WQPIU is a compact stand-alone unit which measures sample induced phase delay under white-light illumination. It does not require any modification of the microscope or additional accessories for its use. The principle of the WQPIU based on lateral shearing interferometry and phase shifting interferometry provides a cost-effective and user-friendly use of QPI. The validity and capacity of the presented method are demonstrated by measuring quantitative phase images of polystyrene beads, human red blood cells, HeLa cells and mouse white blood cells. With speckle-free imaging capability due to the use of white-light illumination, the WQPIU is expected to expand the scope of QPI in biological sciences as a powerful but simple imaging tool.

physics.optics

Single-shot optical imaging through scattering medium using digital in-line holography

Non-invasive and single-shot holographic imaging through complex media is technically challenging due to random light scattering which significantly scrambles optical information. Recently, several methods have been presented to address this issue. However, they require complicated measurements of optical transmission matrices, or existing techniques do only retrieve intensity information. Here we propose and experimentally demonstrate a holographic approach for single-shot imaging through a scattering layer based on digital in-line holography in combination with the autocorrelation of the speckle intensity. Using a simple optical configuration and experimental procedure, the proposed method enables to retrieve the complex amplitude image of an object located at arbitrary planes behind scattering media. The technique has potential applications in biomedical imaging, deep tissue microscopy, and 3D imaging through turbid media.

physics.optics

Optical diffraction tomography techniques for the study of cell pathophysiology

Three-dimensional imaging of biological cells is crucial for the investigation of cell biology, provide valuable information to reveal the mechanisms behind pathophysiology of cells and tissues. Recent advances in optical diffraction tomography (ODT) have demonstrated the potential for the study of various cells with its unique advantages of quantitative and label-free imaging capability. To provide insight on this rapidly growing field of research and to discuss its applications in biology and medicine, we present the summary of the ODT principle and highlight recent studies utilizing ODT with the emphasis on the applications to the pathophysiology of cells.

physics.bio-ph

Label-free optical quantification of structural alterations in Alzheimer's disease

We present a quantitative label-free imaging of mouse whole brain tissue slices with sub-micrometre resolution, employing holographic microscopy and an automated scanning platform. From the measured light field images, scattering coefficients and anisotropies are quantitatively retrieved, which enables access to structural information about brain tissues. As a proof of principle, we demonstrate that these scattering parameters enable us to quantitatively address structural alteration in the brain tissues of mice with Alzheimer's disease.

physics.med-ph

Optical diffraction tomography using a digital micromirror device for stable measurements of 4-D refractive index tomography of cells

Optical diffraction tomography (ODT) is an interferometric microscopy technique capable of measuring 3-D refractive index (RI) distribution of transparent samples. Multiple 2-D holograms of a sample illuminated with various angles are measured, from which 3-D RI map of the sample is reconstructed via the diffraction theory. ODT has been proved as a powerful tool for the study of biological cells, due to its non-invasiveness, label-free and quantitative imaging capability. Recently, our group has demonstrated that a digital micromirror device (DMD) can be exploited for fast and precise control of illumination beams for ODT. In this work, we systematically study the precision and stability of the ODT system equipped with a DMD and present measurements of 3-D and 4-D RI maps of various types of live cells including human red blood cells, white blood cells, hepatocytes, and HeLa cells. Furthermore, we also demonstrate the effective visualization of 3-D RI maps of live cells utilizing the measured information about the values and gradient of RI tomograms.

physics.optics

Large-scale optical diffraction tomography for inspection of optical plastic lenses

Herein is presented an optical diffraction tomography (ODT) technique for measuring 3-D refractive index (RI) maps of optical plastic lenses. A Mach-Zehnder interferometer was used to measure multiple complex optical fields of a plastic lens immersed in RI matching oil, at various rotational orientations. From this, ODT was used to reconstruct a 3-D RI distribution of the plastic lens with unprecedented RI sensitivity (dn = 4.21 x 10^-5) and high resolution (12.8 um). As a demonstration, 3-D RI distributions of a 2-mm-diameter borosilicate sphere and a 5-mm-diameter plastic lens

physics.optics

Hyperspectral optical diffraction tomography

Here, we present a novel microscopic technique for measuring wavelength-dependent three-dimensional (3-D) distributions of the refractive indices (RIs) of microscopic samples in the visible wavelengths. Employing 3-D quantitative phase microscopy techniques with a wavelength-swept source, 3-D RI tomograms were obtained in the range of 450 - 700 nm with a spectral resolution of a few nanometers. The capability of the technique was demonstrated by measuring the hyperspectral 3-D RI tomograms of polystyrene beads, human red blood cells, and hepatocytes. The results demonstrate the potential for label-free molecular specific 3-D tomography of biological samples.

physics.optics

Alterations in cell surface area and deformability of individual human red blood cells in stored blood

The functionality and viability of stored human red blood cells (RBCs) is an important clinical issue in transfusion. To systematically investigate changes in stored whole blood, the hematological properties of individual RBCs were quantified in blood samples stored for various periods with and without a preservation solution called CPDA-1. With 3-D quantitative phase imaging techniques, the optical measurements of the 3-D refractive index (RI) distributions and membrane fluctuations were done at the individual cell level. From the optical measurements, the morphological (volume, surface area and sphericity), biochemical (hemoglobin content and concentration), and mechanical parameters (dynamic membrane fluctuation) were simultaneously quantified to investigate the functionalities and their progressive alterations in stored RBCs. Our results show that the stored RBCs without CPDA-1 had a dramatic morphological transformation from discocytes to spherocytes within 2 weeks which was accompanied with significant decreases in cell deformability and cell surface area, and increases in sphericity. However, the stored RBCs with CPDA-1 maintained their morphology and deformability for up to 6 weeks.

physics.med-ph

Active illumination using a digital micromirror device for quantitative phase imaging

We present a powerful and cost-effective method for active illumination using a digital micromirror device (DMD) for quantitative phase imaging techniques. Displaying binary illumination patterns on a DMD with appropriate spatial filtering, plane waves with various illumination angles are generated and impinged onto a sample. Complex optical fields of the sample obtained with various incident angles are then measured via Mach-Zehnder interferometry, from which a high-resolution two-dimensional synthetic aperture phase image and a three-dimensional refractive index tomogram of the sample are reconstructed. We demonstrate the fast and stable illumination control capability of the proposed method by imaging colloidal spheres and biological cells, including a human red blood cell and a HeLa cell.

physics.optics

Quantitative Morphological and Biochemical Studies on Human Downy Hairs using 3-D Quantitative Phase Imaging

This study presents the morphological and biochemical findings on human downy arm hairs using 3-D quantitative phase imaging techniques. 3-D refractive index tomograms and high-resolution 2-D synthetic aperture images of individual downy arm hairs were measured using a Mach-Zehnder laser interferometric microscopy equipped with a two-axis galvanometer mirror. From the measured quantitative images, the biochemical and morphological parameters of downy hairs were non-invasively quantified including the mean refractive index, volume, cylinder, and effective radius of individual hairs. In addition, the effects of hydrogen peroxide on individual downy hairs were investigated.

physics.bio-ph

3-D refractive index tomograms and deformability of individual human red blood cells from cord blood of newborn infants and maternal blood

Red blood cells (RBCs) from the cord blood of newborn infants have distinctive functions for fetal and infant development. To systematically investigate the biophysical characteristics of individual cord RBCs in newborn infants, a comparative study was performed of RBCs from cord blood of newborn infants, and of adult RBCs from mothers or non-pregnant women, employing optical holographic micro-tomography. Optical measurements of 3-D refractive index distributions, and of dynamic membrane fluctuations of individual RBCs, enabled retrieval of the morphological, biochemical, and mechanical properties of cord, maternal, and adult RBCs at the individual cell level. The volume and surface area of the cord RBCs were significant larger than those of RBCs from non-pregnant women, and cord RBCs have more flattened shapes than RBCs in adults. In addition, the Hb content in the cord RBCs of newborns was significantly greater. The Hb concentration in cord RBCs was higher than for non-pregnant women or maternal RBCs, but they were within the physiological range of adults. Interestingly, the amplitude of dynamic membrane fluctuations in cord RBCs were comparable to those in non-pregnant women and maternal RBCs, suggesting that the deformability of cord RBCs is similar to that of healthy RBCs in adults.

physics.med-ph