• 한국시뮬레이션학회논문지
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• 제17권4호
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• pp.167-174
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• 2008

공초점 현미경(confocal microscopy) 기술의 적용은 살아있는 세포를 고배율로 관찰하는 것을 가능하게 하였다. 알츠하이머나 파킨슨 질환 같은 퇴행성 뇌질환의 경우 뇌세포의 수상돌기의 형태학적 변화가 연관되어 있음이 알려져 있다. 따라서 공초점 현미경 영상으로부터 이러한 정보를 추출하는 방법에 대한 연구가 필요하다. 그러나 공초점 현미경 영상은 명암도 분포가 고르지 않고, 구조의 경계 부분의 번짐 현상 등으로 인해 구조 추출에 어려움을 겪고 있는 실정이다. 따라서 이러한 문제를 극복하고 관심 구조에 대한 특성을 추출할 수 있는 영상처리 기법이 필요하다. 본 논문에서는 뇌세포의 수상돌기 공초점 현미경 사진으로부터 구조정보를 추출하는 새로운 방법을 제안한다. 첫째, 미세 분기 구조의 경계를 향상시키는 비선형 확산 필터링을 적용한다. 둘째, 관심구조를 반복적 역치 선택 방법을 이용해 분할한다. 셋째, 분할된 구조의 분석을 위해 구조의 중심축과 경계선을 추출하기 위한 패스트 마칭 방법(Fast Marching Method)에 기반을 둔 골격화를 수행한다. 본 논문에서 제안된 방법은 기존의 방법들과는 달리 주변 잡음에 덜 민감하였으며 거친 경계선에 영향을 훨씬 적게 받음으로써 보다 정확하고 사실적인 중심축 추출 결과를 보였다.

• Journal of the Optical Society of Korea
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• 제14권1호
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• pp.42-48
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• 2010

We used video-rate Confocal Laser Scanning Microscopy (CLSM) to observe the motion of blood cells in a micro-channel. Video-rate CLSM allowed us to acquire images at the rate of 30 frames per second. The acquired images were used to perform Particle Image Velocimetry (PIV), thus providing the velocity profile of the blood in a micro-channel. While previous confocal microscopy-assisted PIV required exogenous micro/nano particles as the tracing particles, we employed blood cells as tracing particles for the CLSM in the reflection mode, which uses light back-scattered from the sample. The blood flow at various depths of the micro-channel was observed by adjusting the image plane of the microscope. The velocity profile at different depths of the channel was measured. The confocal micro-PIV technique used in the study was able to measure blood velocity up to a few hundreds ${\mu}m/sec$, equivalent to the blood velocity in the capillaries of a live animal. It is expected that the technique presented can be applied for in vivo blood flow measurement in the capillaries of live animals.

• Applied Microscopy
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• 제51권
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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.

• 한국광학회지
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• 제8권4호
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• pp.255-259
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• 1997

본 연구에서는, Nipkow 원판을 사용하는 공초점 현미겨을 구성하여 시료의 2차원 영상을 얻었으며, 이들을 간단한 알고리즘으로 연결하여 표면의 3차원 영상을 얻었다. 마이크로 필름으로 제작한 Nipkow 원판의 적절한 경로로 통과하는 빛의 투과율은 0.5% 정도였으며, 따라서 반사율이 높은 물체만을 관찰할 수 있었다. 광원으로는 파장 692.7 nm의 반도체 레이저를 사용하였다. 구성된 현미경으로 전산기용 기억소자 칩 표면에 식각된 회로의 깊이와 거칠기를 관찰할 수 있었따. 재생전용 compact disk(CD)도 표면의 보호막을 제거하였을 때 표면에 있는 pit들의 배열 구조가 관찰되었다.

• 한국복합재료학회:학술대회논문집
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• 한국복합재료학회 2001년도 춘계학술발표대회 논문집
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• pp.201-204
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• 2001

A Confocal Laser Scanning Microscope (CLSM) is applied to determine three-dimensional fiber orientation states in injection-molded short fiber composites. Since the CLSM optically sections the composites, more than two planes either on or below the surface of composites can be obtained. Therefore, three dimensional fiber orientation states are determined without destruction. To predict the orientation states, velocity and temperature fields are calculated by using a hybrid FEM/FDM method. The change of orientation state during packing stage is also considered by employing a compressible Hele-Shaw model. The predicted orientation states show good agreement with measured ones. However, some differences are found at the end of cavity. They may result from other effects, which are not considered in the numerical analysis.

• Archives of Pharmacal Research
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• 제28권1호
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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.

• Fibers and Polymers
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• 제2권1호
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• pp.163-172
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• 2001

To determine three-dimensional fiber orientation states in injection-molded short fiber composites a CLSM (Confocal Laser Scanning Microscope) is used. Since the CLSM optically sections the composites, more than two cross-sections either on or below the surface of the composite can be obtained. Three dimensional fiber orientation states can be determined with geometric parameters of fibers on two parallel cross-sections. For experiment, carbon fiber reinforced polystyrene is examined by the CLSM. Geometric parameters of fibers are measured by image analysis. In order to compactly describe fiber orientation states, orientation tensors are used. Orientation tensors are determined at different positions of the prepared specimen. Three dimensional orientation states are obtained without the difficulty in determining the out-of-plane angles by utilizing images on two parallel planes acquired by the CLSM. Orientation states are different at different positions and show the shell-core structure along the thickness of the specimen.

• 한국레이저가공학회:학술대회논문집
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• 한국레이저가공학회 2006년도 춘계학술발표대회 논문집
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• pp.123-128
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• 2006

We constructed a micro-patterning system that build patterns on a photoresist coated wafer using laser direct writing system. Confocal microscope system was adapted for real-time auto-focusing of the laser writing lens to generate lines of uniform width.

• Journal of the Optical Society of Korea
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• 제12권2호
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• pp.107-111
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• 2008

We demonstrated single-molecule fluorescence resonance energy transfer (FRET) from single donor-acceptor dye pair attached to a DNA with a setup based on a confocal microscope. Singlestrand DNAs were immobilized on a glass surface with suitable inter-dye distance. Energy transfer efficiency between the donor and the acceptor dyes attached to the DNA was measured with different lengths of DNA. Photobleaching of single dye molecule was observed and used as a sign of single-molecule detection. We could achieve high enough signal-to-noise ratio to detect the fluorescence from a single-molecule, which allows real-time observation of the distance change between single dye pairs in nanometer scale.

• Journal of the Optical Society of Korea
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• 제9권1호
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• pp.32-35
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• 2005

In light microscopy, spatial resolution is limited by diffraction effect. Confocal microscopy has improved resolutions in both lateral and axial directions, but these are still limited by diffraction effect. Confocal self-interference microscopy (CSIM) uses interference between two perpendicularly polarized beams to enhance lateral resolution. In previous research, we proposed a calcite plate with its optic-axis perpendicular to the propagation angle and one of the boundary surfaces of the plate. This type of plate is not widely used to our knowledge. In this paper, we change the calcite plate to more common one, which is commercially available. This calcite plate has its optic axis in the plane of incidence. We analyze the characteristics of this calcite plate and numerically compare the performances of CSIM in previous research and CSIM with the commercial calcite plate. Numerical results show improved performance when using the commercial calcite plate