• 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.

• Proceedings of the Korean Society of Precision Engineering Conference
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• 2005.10a
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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.

• Current Optics and Photonics
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• v.4 no.1
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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.

• Korean Journal of Optics and Photonics
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• v.20 no.4
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• pp.207-210
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• 2009

A confocal technique was demonstrated for measuring the absolute value of an air gap between substrates. Since the two surfaces were in Rayleigh range of the laser focus for air gaps less than 200 nm, complete interference patterns were observed. And since interference patterns were obtained only from the area of focus, it was an advantage of this method that air gaps between multiple thin films could be measured. Stability is less than 1 nm except in the range where the interference pattern changes slowly.

• Journal of the Korea Society for Simulation
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• v.17 no.4
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• pp.167-174
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• 2008

The introduction of confocal microscopy makes it possible to observe the structural change of live neuronal cell. Neuro-degenerative disease, such as Alzheimer;s and Parkinson’s diseases are especially related to the morphological change of dendrite spine. That’s the reason for the study of segmentation and extraction from confocal microscope image. The difficulty comes from uneven intensity distribution and blurred boundary. Therefore, the image processing technique which can overcome these problems and extract the structural information should be suggested. In this paper, we propose robust structural information extracting technique with confocal microscopy images of dendrite in brain neurons. First, we apply the nonlinear diffusion filtering that enhance the boundary recognition. Second, we segment region of interest using iterative threshold selection. Third, we perform skeletonization based on Fast Marching Method that extracts centerline and boundary for analysing segmented structure. The result of the proposed method has been less sensitive to noise and has not been affected by rough boundary condition. Using this method shows more accurate and objective results.

• Journal of the Optical Society of Korea
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• v.14 no.1
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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.

• Journal of the Korean Society of Visualization
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• v.8 no.1
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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.

• Korean Journal of Clinical Laboratory Science
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• v.46 no.4
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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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• 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.