• Medical Lasers
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• 제10권1호
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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.

• 반도체디스플레이기술학회지
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• 제7권1호
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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.

• 한국정밀공학회:학술대회논문집
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• 한국정밀공학회 2005년도 추계학술대회 논문집
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• pp.576-580
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• 2005

New real-time confocal microscopy using spectral encoding technique and slit confocal aperture is proposed and designed. Spectral encoding technique, which encodes one-dimensional spatial information of a specimen in wavelength, and slit aperture make it possible to obtain two-dimensional lateral image of the specimen simultaneously at standard video rates without expensive scanning units such as polygon mirrors and galvano mirrors. The working principle and the configuration of the system are explained. The variation in axial responses for the simplified model of the system with normalized slit width is numerically analyzed based on the wave optics theory. Slit width that directly affects the depth discrimination of the system is determined by a compromise between axial resolution and signal intensity from the simulation result. On the assumption of the lateral sampling resolution of 50 nm, design variables and governing equations of the system are derived. The system is designed to have the mapping error less than the half pixel size, to be diffraction-limited and to have the maximum illumination efficiency. The designed system has the FOV of $12.8um{\times}9.6um$, the theoretical axial FWHM of 1.1 um and the lateral magnification of-367.8.

• 한국광학회지
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• 제20권4호
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• pp.207-210
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• 2009

공초점 방식을 이용하여 두 기판사이의 얇은 공기갭의 두께를 측정하는 방법을 제안하였다. 공기갭이 약 200 nm 이내 일때는 두 기판면이 초점의 레일리 영역 안에 있으므로 완전한 간섭신호를 측정할 수 있었다. 또 초점영역 근처에서의 간섭무늬만 측정되므로 다층 박막중에 존재하는 공기갭이라도 측정할 수 있는 장점이 있다. 간섭무늬 변화가 작은 영역을 제외하면 1 nm 이내의 안정도를 가진다.

• Current Optics and Photonics
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• 제4권1호
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• pp.44-49
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• 2020

In this paper, a new method is presented to measure the refractive index of single plain glass or multilayered materials, based on a laser frequency-shifted confocal feedback microscope. Combining the laser frequency-shifted feedback technique and the confocal effect, the method can attain high axial-positioning accuracy, stability and sensitivity. Measurements of different samples are given, including N-BK7 glass, Silica plain glass, and a microfluidic chip with four layers. The results for N-BK7 glass and Silica plain glass show that the measurement uncertainty in the refractive index is better than 0.001. Meanwhile, the feasibility of this method for multilayered materials is tested. Compared to conventional methods, this system is more compact and has less difficulty in sample processing, and thus is promising for applications in the area of refractive-index measurement.

• 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.

• 한국가시화정보학회지
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• 제8권1호
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• pp.46-52
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• 2010

In the present study, we employed the confocal laser scanning microscopy (CLSM) system to visualize the blood flow field with $1{\times}1{\mu}m^2$ spatial resolution. Based on the confocal microscopic image of red blood cells (RBCs), we performed the velocity vector field measurement and evaluated characteristics of cell migration from the cell depleted layer thickness calculation. The rat and mouse's blood were supplied into a micro glass tubes in vitro. The line scanning rate of confocal microscopy was 15 kHz for a $500{\times}500$ pixels image. As a result, the red blood cell itself can be used as a tracer directly without any kind of invasive tracer particle to get the velocity vector field of blood flow by performing particle image velocimetry (PIV) technique.

• 대한임상검사과학회지
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• 제46권4호
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• pp.136-139
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• 2014

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the severe nosocomial infectious agents. The traditional diagnostic methods including biochemical test, antibiotic susceptibility test and PCR amplification are time consuming and require much work. The Surface enhanced Raman spectroscopy (SERS) biosensor is a rapid and powerful tool for analyzing the chemical composition within a single living cell. To identify the biochemical and genetic characterization of clinical MRSA, all isolates from patients were performed with VITEK2 gram positive (GP) bacterial identification and Antibiotic Susceptibility Testing (AST). Virulence genes of MRSA also were identified by DNA based PCR using specific primers. All isolates, which were placed on a gold coated nanochip, were analyzed by a confocal Raman microscopy system. All isolates were identified as S. aureus by biochemical tests. MRSA, which exhibited antibiotic resistance, demonstrated to be positive gene expression of both femA and mecA. Furthermore, Raman shift of S. aureus and MRSA (n=20) was perfectly distinguished by a confocal Raman microscopy system. This novel technique explained that a SERS based confocal Raman microscopy system can selectively isolate MRSA from non-MRSA. The study recommends the SERS technique as a rapid and sensitive method to detect antibiotic resistant S. aureus in a single cell level.

• Current Optics and Photonics
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• 제2권6호
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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.