• 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.

• Archives of Pharmacal Research
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• v.28 no.1
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• pp.93-99
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• 2005

Intracellular trafficking of transferrin-conjugated dimethyldioctadecyl-ammonium bromide liposome $(T_f-liposome)/DNA$ complexes in HeLa cells was studied using the double-labeled fluorescence technique and confocal microscopy. The size of the $T_f-liposome/DNA$ complex was about 367 nm in diameter and the zeta-potential of it at a 5:1 (w/w) ratio was almost neutral. The intracellular pathway of the $T_f-liposome/DNA$ complex, noted as green (FITC), red (rhodamine) or yellow (FITC + rhodamine) fluorescence, was elucidated from the plasma membrane to the endosome (or lysosome), and finally to the nucleus. The results of this study indicate that plasmid DNA enters into the nucleus not only as a free form but as an associated form complexed with $T_f-liposome$. More interestingly, the $T_f-liposome$ undergoes a nuclear location in the form of ordered structures. This could be a very useful piece of information in designing a safe and advanced gene delivery system.

• International Journal of Precision Engineering and Manufacturing
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• v.7 no.4
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• pp.3-7
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• 2006

Confocal scanning microscopy is a measurement technique used to observe micrometer and sub-micrometer features due to its high resolution, nondestructive properties, and 3D surface profiling capabilities. The design, implementation, and performance test of a confocal scanning microscopy system are presented in this paper. A short-wavelength laser (405 nm) and an objective lens with a high numerical aperture (0.95) were used to achieve the desired high resolution, while the x- and y-axis scans were implemented using an acousto-optic deflector and galvanomirror, respectively. An objective lens with a piezo-actuator was used to scan the z-axis. A spatial resolution of less than 138 nm was achieved, along with successful 3D surface reconstructions.

• Journal of the Korean Society of Manufacturing Technology Engineers
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• v.21 no.5
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• pp.708-713
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• 2012

The indented geometry for rockwell hardness indenter has been configured by using Confocal Laser Scanning Microscopy (CLSM). For this purpose, the CLSM can be well suited to construct the three-dimensional indented volume from the indented surface by rockwell hardness tester. Furthermore, the height data of HEI(height encoded image) by CLSM must be acquired at first and converted to indented surface later. And the indented surface patterns enable us to predict the indenter shape and volume. This volume can be used to study the rockwell hardness model as a volume parameter. As a result, the technique performed in this study by combining the CLSM with compensation technique is an excellent one to obtain the geometries of indented surfaces over a wide range of surface resolution in a micro scale. And it can be used for micro volume calculation.

• Korean Journal of Optics and Photonics
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• v.15 no.6
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• pp.583-590
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• 2004

The autofocus is one of the important processes in the automated vision inspection or measurements using optical microscopes, because it influences the measuring accuracy. In this paper, we used the confocal microscope configuration based on not a pinhole but a single-mode optical fiber. A single mode fiber has the functions of source and detector by applying the reciprocal scheme. As a result, we acquired a simple system configuration and easy alignment of the optical axis. Also, we embodied a fast autofocus system by acquiring the focus error signal through a source modulation technique. The source modulation technique can effectively reduce physical disturbances compared with objective lens modulation, and it is easily applicable to general optical microscopes. The focus error signal was measured with respect to the modulation amplitude, reflectance of the specimen and inclination angle of the measuring surface. The performance of the proposed autofocus system was verified through autofocusing flat mirror surface. In addition, we confirmed that source modulation rarely degrades the depth resolution by the comparison between the FWHMs of axial response curves.

• 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.

• 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.

• Korean Journal of Optics and Photonics
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• v.13 no.2
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• pp.155-161
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• 2002

The nail bed is located under the finger nail. The arched portions of the nail bed, which contain a large number of capillary loops, are separated by the valley of the nail bed. The valley of the nail bed does not contain capillary loops. Light is scattered when it propagates through the dermis of skin, and human blood strongly absorbs the light with proper wavelength. By use of the optical properties of the nail bed, we propose an optical technique which extracts the nail bed image of the finger nail. After achieving nail bed images of each individual, we correlated between them. The correlation outputs show that we can identify individuals by comparing the peak heights of the correlation outputs.

• 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.

• Clinical Endoscopy
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• v.57 no.4
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• pp.434-445
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• 2024

Pancreatic cystic lesions (PCLs) have increased in prevalence due to the increased usage and advancements in cross-sectional abdominal imaging. Current diagnostic techniques cannot distinguish between PCLs requiring surgery, close surveillance, or expectant management. This has increased the morbidity and healthcare costs from inappropriately aggressive and conservative management strategies. Endoscopic ultrasound (EUS) needle-based confocal laser endomicroscopy (nCLE) allows for microscopic examination and delineation of the surface epithelium of PCLs. Landmark studies have identified characteristics distinguishing various types of PCLs, confirmed the high diagnostic yield of EUS-nCLE (especially for PCLs with an equivocal diagnosis), and shown that EUS-nCLE helps to change management and reduce healthcare costs. Refining procedure technique and reducing procedure length have improved the safety of EUS-nCLE. The utilization of artificial intelligence and its combination with other EUS-based advanced diagnostic techniques would further improve the results of EUS-based PCL diagnosis. A structured training program and device improvements to allow more complete mapping of the pancreas cyst epithelium will be crucial for the widespread adoption of this promising technology.