• Journal of the Optical Society of Korea
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• v.15 no.3
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• pp.300-304
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• 2011

Optical microscopes are widely used for medical imaging these days, but biopsy is a lengthy process that causes many problems during the ex-vivo imaging procedure. The endo-microscope has been studied to increase accessibility to the human body and to get in-vivo images to use for medical diagnosis. This research proposes a multi-modal confocal endo-microscope for bio-medical imaging. We introduce the design process for a small endoscopic probe and a coupling mechanism for the probe to make the multi-modal confocal endo-microscope. The endoscopic probe was designed to decrease chromatic and spherical aberrations, which deteriorate the images obtained with the conventional GRIN lens. Fluorescence and reflectance images of various samples were obtained with the proposed endo-microscope. We evaluated the performance of the proposed endo-microscope by analyzing the acquired images, and demonstrate the possibilities of in-vivo medical imaging for early diagnosis.

• Proceedings of the KOSOMBE Conference
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• v.1995 no.11
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• pp.257-261
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• 1995

Confocal laser scanning microscopy (CLSM)는 기존의 coherent or incoherent microscopic imaging 보다 횡축 방향 (lateral direction)으로 고해상도를 가지며, 층과 층 사이를 구분하는 광축 방향 (axial direction)의 optical sectioning에 의해 샘플의 3D 구조를 고해상도로 영상화함으로써 3D 구조 및 생체 기능 분석을 가능하게 해 준다. 본 논문에서는 CLSM에 의한 3D 영상화 원리와 촛점면 부근에서 얻어지는 광세기 분포, 얻어진 2D slice 영상의 시각화 및 응용에 대해 논의된다.

• Applied Microscopy
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• v.51
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• pp.4.1-4.12
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• 2021

Fluorescence in situ hybridization (FISH) is a technique to visualize specific DNA/RNA sequences within the cell nuclei and provide the presence, location and structural integrity of genes on chromosomes. A confocal Whole Slide Imaging (WSI) scanner technology has superior depth resolution compared to wide-field fluorescence imaging. Confocal WSI has the ability to perform serial optical sections with specimen imaging, which is critical for 3D tissue reconstruction for volumetric spatial analysis. The standard clinical manual scoring for FISH is labor-intensive, time-consuming and subjective. Application of multi-gene FISH analysis alongside 3D imaging, significantly increase the level of complexity required for an accurate 3D analysis. Therefore, the purpose of this study is to establish automated 3D FISH scoring for z-stack images from confocal WSI scanner. The algorithm and the application we developed, SHIMARIS PAFQ, successfully employs 3D calculations for clear individual cell nuclei segmentation, gene signals detection and distribution of break-apart probes signal patterns, including standard break-apart, and variant patterns due to truncation, and deletion, etc. The analysis was accurate and precise when compared with ground truth clinical manual counting and scoring reported in ten lymphoma and solid tumors cases. The algorithm and the application we developed, SHIMARIS PAFQ, is objective and more efficient than the conventional procedure. It enables the automated counting of more nuclei, precisely detecting additional abnormal signal variations in nuclei patterns and analyzes gigabyte multi-layer stacking imaging data of tissue samples from patients. Currently, we are developing a deep learning algorithm for automated tumor area detection to be integrated with SHIMARIS PAFQ.

• Development and Reproduction
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• v.21 no.3
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• pp.229-235
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• 2017

In the field of reproductive medicine, assessment of sperm motility is a key factor for achieving successful artificial insemination, in vitro fertilization, or intracellular sperm injection. In this study, the motility of boar sperms was estimated using real-time imaging via confocal microscopy. To confirm this confocal imaging method, flagellar beats and whiplash-like movement angles were compared between fresh and low-temperature-preserved ($17^{\circ}C$ for 24 h) porcine sperms. Low-temperature preservation reduced the number of flagellar beats from $11.0{\pm}2.3beats/s$ (fresh sperm) to $5.7{\pm}1.8beats/s$ and increased the flagellar bending angle from $19.8^{\circ}{\pm}13.8^{\circ}$ (fresh) to $30.6^{\circ}{\pm}15.6^{\circ}$. These data suggest that sperm activity can be assessed using confocal microscopy. The observed motility patterns could be used to develop a sperm evaluation index and automated confocal microscopic sperm motility analysis techniques.

• Medical Lasers
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• v.10 no.1
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• pp.1-6
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• 2021

Intravital microscopy is a high-resolution imaging technique based on laser-scanning two-photon and confocal microscopy, which allows dynamic 3D cellular-level imaging of various biological processes in a living animal in vivo. This unique capability allows biomedical researchers to directly verify a hypothesis in a natural in vivo microenvironment at the cellular level in a physiological setting. During the last decade, intravital microscopy has become an indispensable technique in several fields of biomedical sciences such as molecular and cell biology, immunology, neuroscience, developmental, and tumor biology. The most distinct advantage of intravital microscopy is its capability to provide a longitudinal view of disease progression at the cellular-level with repeated intravital imaging of a single animal over time by saving the images after each session.

• Journal of the Semiconductor & Display Technology
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• v.7 no.1
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• pp.11-16
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• 2008

Confocal scanning microscopy is a widely used technique for three dimensional measurements because it is characterized by high resolution, high SNR and depth discrimination. Generally an image is generated by moving one optical probe that satisfies the confocal condition on the specimen. Measurement speed is limited by movement speed of the optical probe; scanning speed. To improve measurement speed we increase the number of optical probes. Specimen region to scan is divided by optical probes. Multi-point information each optical probe points to can be obtained simultaneously. Therefore image acquisition speed is increased in proportion to the number of optical probes. And multiple optical probes from red, green and blue laser sources can be used for color imaging and image quality, i.e., contrast, is improved by adding color information by this way. To conclude, this technique contributes to the improvement of measurement speed and image quality.

• Journal of the Optical Society of Korea
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• v.15 no.1
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• pp.61-67
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• 2011

The confocal laser scanning microscopy and two-photon microscopy was implemented based on a single laser source and an objective lens. We imaged and compared the morphology of identical sites of ex vivo human skin using both microscopes. The back-scattering emission from the sample provided the contrast for the confocal microscopy. The intrinsic autofluorescence and the second harmonic generation were used as the luminescence source for the two-photon microscopy. The wavelength of the Ti:Sapphire laser was tuned at 710 nm, which corresponds to the excitation peak of NADH and FAD in skin tissue. The various cell layers in the epidermis and the papillary dermis were clearly distinguished by both imaging modalities. The two-photon microscopy more clearly visualized the intercellular region and the nucleus of the cell compared to the confocal microscopy. The fibrous structures in the dermis were more clearly resolved by the confocal microscopy. Numerous cells in papillary dermal layer, as deep as $100\;{\mu}m$, were observed in both CLSM and two-photon microscopy. While most previous studies focused on fibrous structure imaging (collagen and elastin fiber) in the dermis, we demonstrated that the combined imaging with the CLSM and two-photon microscopy can be applied for the non-invasive study of the population, distribution and metabolism of papillary dermal cells in skin.

• Journal of Institute of Control, Robotics and Systems
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• v.21 no.12
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• pp.1130-1135
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• 2015

The bond pull test is the most widely used technique for the evaluation and control of wire bond quality. The wire being tested is pulled upward until the wire or bond to the die or substrate breaks. The inspector test strength of wire by manually and it takes around 3 minutes to perform the test. In this paper, we develop a 3D vision system to measure 3D position of wire. It gives 3D position data of wire to move a hook into wires. The 3D measurement method to use here is a confocal imaging system. The conventional confocal imaging system is a spot scanning method which has a high resolution and good illumination efficiency. However, a conventional confocal systems has a disadvantage to perform XY axis scanning in order to achieve 3D data in given FOV (Field of View) through spot scanning. We propose a method to improve a parallel mode confocal system using a micro-lens and pin-hole array to remove XY scan. 2D imaging system can detect 2D location of wire and it can reduce time to measure 3D position of wire. In the experimental results, the proposed system can measure 3D position of wire with reasonable accuracy.

• Current Optics and Photonics
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• v.2 no.6
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• pp.568-575
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• 2018

Various techniques to measure the three-dimensional (3D) surface profile of a 3D micro- or nanostructure have been proposed. However, it is difficult to apply such techniques directly to industrial uses because most of them are relatively slow, unreliable, and expensive. The HiLo optical imaging technique, which was recently introduced in the field of fluorescence imaging, is a promising wide-field imaging technique capable of high-speed imaging with a simple optical configuration. It has not been used in measuring a 3D surface profile although confocal microscopy originally developed for fluorescence imaging has been adapted to the field of 3D optical measurement for a long time. In this paper, to the best of our knowledge, the HiLo optical imaging technique for measuring a 3D surface profile is proposed for the first time. Its optical configuration and algorithm for a precisely detecting surface position are designed, optimized, and implemented. Optical performance for several 3D microscale structures is evaluated, and it is confirmed that the capability of measuring a 3D surface profile with HiLo optical imaging technique is comparable to that with confocal microscopy.

• 한국가시화정보학회:학술대회논문집
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• 2006.12a
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• pp.96-101
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• 2006

3-D visualization using confocal laser scanning microscopy (CLSM) in a chaotic micromixer was performed as a reproduction experiment and the feasibility of 3-0 imaging technique in the microscale was confirmed. For diagonal micromixer (DM) and two types of staggered herringbone micromixers (SHM) designed by Whitesides et al., to verify the evolution of mixing, cross sectional images are reconstructed at the end of every cycle. In a DM, clockwise rotational flow motion generated by diagonal ridges placed on the floor of micromixer is observed and this motion makes the fluid commingle. On the contrary, there are two rotational flow structures in the SHM and the centers of rotation exchange their position each other every half cycle because of the V shape of ridges varying their orientation every half cycle. Local rotational flow and local extensional flow generated by the complicate ridge pattern make the flow be chaotic and accelerate the mixing of fluid. The dominant parameter that influences on the mixing characteristic of SHM is not the length of micromixer but the number of ridges under the same flow configurations.