• Title/Summary/Keyword: Optical sectioning

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Flexure hinge mechanism having amplified rectilinear motion for confocal scanning microscopy using optical section

  • Kwon, Oh-Kyu;Park, Poo-Gyeon
    • 제어로봇시스템학회:학술대회논문집
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    • 2001.10a
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    • pp.162.6-162
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    • 2001
  • Confocal scanning microscopy (CSM) is an important instrument in a wide variety of imaging applications because of its ability to provide three-dimensional images of thick, volume specimens. The mechanism for two-dimensional beam scanning and optical sectioning has an important roe in CSM as the three-dimensional profiler. This optical sectioning property arises from the use of a point detector, which serves to attenuate the signals from out-of-focus. The intensity profile for the open loop scanning should be matched with its response for the standard. The non-linearity can be minimized with the optical sectioning or the optical probe of the closed loop control. This paper shows the mathematical expression of the light such as the extinction curve in the optical fields of system using AO deflector, the axial/lateral response experimentally when the error sources change, and the methods of optical sectioning. Thorough design of optical sectioner is crucial to the success of CSM in the field ...

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Optical Tracking of Three-Dimensional Brownian Motion of Nanoparticles

  • Choi C. K.;Kihm K.D.
    • Journal of the Korean Society of Visualization
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    • v.3 no.1
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    • pp.3-19
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    • 2005
  • Novel optical techniques are presented for three-dimensional tracking of nanoparticles; Optical Serial Sectioning Microscopy (OSSM) and Ratiometric Total Internal Reflection Fluorescent Microscopy (R-TIRFM). OSSM measures optically diffracted particle images, the so-called Point Spread Function (PSF), and dotermines the defocusing or line-of-sight location of the imaged particle measured from the focal plane. The line-of-sight Brownian motion detection using the OSSM technique is proposed in lieu of the more cumbersome two-dimensional Brownian motion tracking on the imaging plane as a potentially more effective tool to nonintrusively map the temperature fields for nanoparticle suspension fluids. On the other hand, R-TIRFM is presented to experimentally examine the classic theory on the near-wall hindered Brownian diffusive motion. An evanescent wave field from the total internal reflection of a 488-nm bandwidth of an argon-ion laser is used to provide a thin illumination field of an order of a few hundred nanometers from the wall. The experimental results show good agreement with the lateral hindrance theory, but show discrepancies from the normal hindrance theory. It is conjectured that the discrepancies can be attributed to the additional hindering effects, including electrostatic and electro-osmotic interactions between the negatively charged tracer particles and the glass surface.

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Proposal and design of reflecting optical system to improve detection intensity in fluorescence confocal scanning microscopy (형광 공초점 주사 현미경의 측정 강도 향상을 위한 반사 광학계의 제안 및 설계)

  • 강동균;서정우;권대갑
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 2002.05a
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    • pp.187-190
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    • 2002
  • Confocal microscopy is very popular technology in bio-medical inspection due to its ability to reject background signals and to measure very thin slide of thick specimens, which is called optical sectioning. But intensity of detected signal in fluorescence type confocal microscopy is so small that only 0.2% of emitted fluorescence light can be detected in the best case. In this paper, we proposed the reflecting optical system to improve the detection intensity and designed the optical system by optimal design method. At the end of the paper, we analyzed the characteristics of the proposed reflecting optical system.

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Study on the Contamination of Pseudomonas tolaasii in Oyster Mushroom (느타리버섯에서의 Pseudomonas tolaasii오염 연구)

  • 이혜영;장금일;김광엽
    • Journal of Food Hygiene and Safety
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    • v.16 no.3
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    • pp.232-240
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    • 2001
  • One hundred twenty five bacterial isolates were obtained from the brown blotch-diseased oyster mushrooms collected from markets. Among them, 45 were determined as pathogenic bacteria and white line forming organisms(WLFO) were 6 strains and white line reaction organisms (WLRO) were 6 strains. All of the white line forming isolates were identified as Pseudomonas tolaasii which is a known pathogen of brown blotch disease of oyster mushroom by GC-MIS(Gas chromatography-microbial identification system). Six of the white line reacting organisms were identified as P. chlomraphis, P. fluorescens biotype A and type C. The rest of them were P gingeri, P. agarici, P. fluorescens biotype B, P. chloroyaphis, non-pathogenic P. tolaasii, P. putida biotype A and B etc. For spectrum of activity of tolaasin, culture filtrates from pathogenic isolates were examined by browning of mushroom tissue and pitting of mushroom caps. The weak pathogenic bacteria didn't induce browning or pitting of mushroom tissue. On the other hand, strong pathogenic isolates showed browning and pitting reaction on mushroom. An extracellular toxin produced by P. tolaasii, was investigated. The hemolysis activity test of 6 strains identified as P. tolaasii were 0.8∼0.9 at 600 nm and 3 strains of WLRO were 0.9∼1.0 and Pseudomonas app. were 1.0∼1.2. Observation of fresh mushroom tissue using confocal laser scanning microscopy was carried out for images of optical sectioning and vertical sectioning. Also images of brown blotch diseased oyster mushroom tissue after contamination P. tolaasii was obtained by CLSM.

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BGA Height Measurement Using Pattern Beam (패턴 빔을 이용한 BGA 단차 측정)

  • Shin, Sang-Hoon;Yu, Young-Hun
    • Korean Journal of Optics and Photonics
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    • v.20 no.6
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    • pp.361-365
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    • 2009
  • We describe a simple method to obtain an optical sectioning in a conventional wide-field microscope by projecting a single spatial frequency grid pattern onto the object. Using a patterned beam, we have measured the height of BGA with a rough surface that provide the coherence noise. The configuration of the height measurement system using pattern beam is simple. The image acquired by this system is not depend on the coherence noises. This system is also applicable to the sample reference plan that has no pattern on ground. The reappearance and accuracy are outstanding and applicable to many industrial optical metrology.

Recent Research on Self-interference Incoherent Digital Holography

  • Youngrok Kim;Ki-Hong Choi;Chihyun In;Keehoon Hong;Sung-Wook Min
    • Current Optics and Photonics
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    • v.8 no.1
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    • pp.1-15
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    • 2024
  • This paper presents a brief introduction to self-interference incoherent digital holography (SIDH). Holography conducted under incoherent light conditions has various advantages over digital holography performed with a conventional coherent light source. We categorize the methods for SIDH, which divides the incident light into two waves and modulates them differently. We also explore various optical concepts and techniques for the implementation and advancement of SIDH. This review presents the system design, performance analysis, and improvement of SIDH, as well as recent applications of SIDH, including optical sectioning and deep-learning-based SIDH.

Three Dimensional Confocal Imaging and Biomedical Image Analysis (3차원 Confocal Imaging과 생체 영상 분석)

  • Lee, Yim-Kul
    • 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.

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implementation and its limitations

  • Nahm, Kie-B.;Shin, Eun-S.;Ryoo, Seok-M.
    • Journal of the Optical Society of Korea
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    • v.1 no.2
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    • pp.90-93
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    • 1997
  • The shallow depth of focus in conventional light microscopy hinders the observation of the whole image when the object is thicker than the depth of field. Most of the existing techniques measured the object distance, which is not necessarily the actual distance of each pixel in the image. We implemented a means of determining the "best focus" of each pixel and located the height of object points by sectioning at different sample heights. Combining the height information and its gray values together, we obtained an image where the blur from the finite depth of focus is eliminated. Limitations of the technique are discussed together with composed images.ed images.

High-speed Two-photon Laser Scanning Microscopy Imaging of in vivo Blood Cells in Rapid Circulation at Velocities of Up to 1.2 Millimeters per Second

  • Boutilier, Richard M.;Park, Jae Sung;Lee, Ho
    • Current Optics and Photonics
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    • v.2 no.6
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    • pp.595-605
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    • 2018
  • The two-photon process of microscopy provides good spatial resolution and optical sectioning ability when observing quasi-static endogenous fluorescent tissue within an in vivo animal model skin. In order to extend the use of such systems, we developed a two-photon laser scanning microscopy system capable of also capturing $512{\times}512$ pixel images at 90 frames per second. This was made possible by incorporating a 72 facet polygon mirror which was mounted on a 55 kRPM motor to enhance the fast-scan axis speed in the horizontal direction. Using the enhanced temporal resolution of our high-speed two-photon laser scanning microscope, we show that rapid processes, such as fluorescently labeled erythrocytes moving in mouse blood flow at up to 1.2 mm/s, can be achieved.

Subcellular Location of Spodpotera Cell-expressed Human HepG2-type Glucose Transport Protein

  • Lee, Chong-Kee
    • Biomedical Science Letters
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    • v.18 no.2
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    • pp.160-164
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    • 2012
  • The baculovirus/insect cell expression system is of great value for the large-scale production of normal and mutant mammalian passive glucose-transport proteins heterologously for structural and functional studies. In most mammalian cells that express HepG2, this transporter isoform is predominantly located at the cell surface. However, it had been reported that heterologous expression of other membrane proteins using the baculovirus system induced highly vacuolated cytoplasmic membranes. Therefore, how a cell responds to the synthesis of large amounts of a glycoprotein could be an interesting area for investigation. In order to examine the subcellular location of the human HepG2 transport proteins when expressed in insect cells, immunofluorescence studies were carried out. Insect cells were infected with the recombinant baculovirus AcNPVHIS-GT or with wild-type virus at a MOI of 5, or were not exposed to viral infection. A high level of fluorescence displayed in cells infected with the recombinant virus indicated that transporters are expressed abundantly and present on the surface of infected Sf21 cells. The evidence for the specificity of the immunostaining was strengthened by the negative results shown in the negative controls. Distribution of the transporter protein expressed in insect cells was further revealed by making a series of optical sections through an AcNPVHIS-GT-infected cell using a confocal microscope, which permits optical sectioning of cell sample. These sections displayed intense cytoplasmic immunofluorecence surrounding the region occupied by the enlarged nucleus, indicating that the expressed protein was present not only at the cell surface but also throughout the cytoplasmic membranous structures.