• Title/Summary/Keyword: Confocal microscopy

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High-speed Three-dimensional Surface Profile Measurement with the HiLo Optical Imaging Technique

  • Kang, Sewon;Ryu, Inkeon;Kim, Daekeun;Kauh, Sang Ken
    • 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.

Characteristics of the Infection of Tilletia laevis Kuhn (syn. Tilletia foetida (Wallr.) Liro.) in Compatible Wheat

  • Ren, Zhaoyu;Zhang, Wei;Wang, Mengke;Gao, Haifeng;Shen, Huimin;Wang, Chunping;Liu, Taiguo;Chen, Wanquan;Gao, Li
    • The Plant Pathology Journal
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    • v.37 no.5
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    • pp.437-445
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    • 2021
  • Tilletia laevis Kuhn (syn. Tilletia foetida (Wallr.) Liro.) causes wheat common bunt, which is one of the most devastating plant diseases in the world. Common bunt can result in a reduction of 80% or even a total loss of wheat production. In this study, the characteristics of T. laevis infection in compatible wheat plants were defined based on the combination of scanning electron microscopy, transmission electron microscopy and laser scanning confocal microscopy. We found T. laevis could lead to the abnormal growth of wheat tissues and cells, such as leakage of chloroplasts, deformities, disordered arrangements of mesophyll cells and also thickening of the cell wall of mesophyll cells in leaf tissue. What's more, T. laevis teliospores were found in the roots, stems, flag leaves, and glumes of infected wheat plants instead of just in the ovaries, as previously reported. The abnormal characteristics caused by T. laevis may be used for early detection of this pathogen instead of molecular markers in addition to providing theoretical insights into T. laevis and wheat interactions for breeding of common bunt resistance.

Fiber Orientation in Injection-Molded Short Fiber Composites with a Confocal Laser Scanning Microscope and Numerical Simulation (공초점 레이저 주사 현미경을 이용한 단섬유 복합재료 사출 성형물 내의 섬유 배열 측정 및 수치모사)

  • Lee, Kwang-Seok;Le, Seok-Won;Youn, Jae-Ryoun
    • Proceedings of the Korean Society For Composite Materials Conference
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    • 2001.05a
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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.

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Studies on Salmonella enteritidis Contamination in Chicken Egg using Confocal Scanning Laser Microscopy (Confocal Scanning Laser Microscopy 를 이용한 계란에서의 Salmonella enteritidis 오염 연구)

  • Jang, Keum-Il;Park, Jong-Hyun;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.31 no.3
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    • pp.771-777
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    • 1999
  • The structural function of three egg membrane layers and cuticle layer, and the effectiveness of 5 film coatings (chitosan, starch, gelatin, dextrin, mineral oil) on the prevention of Salmonella enteritidis penetration was investigated using confocal scanning laser microscopy (CSLM). Diameters of outer membrane fibers, inner membrane fibers and limiting membrane particles in eggshell were $1.5{\sim}7.2$, $0.8{\sim}2.0$ and $0.1{\sim}1.4\;{\mu}m$, respectively and average thicknesses were 10.0, 3.5, $3.6\;{\mu}m$, respectively. Average thickness of cuticle layer was $6.0\;{\mu}m$ and cuticle layer covered $40{\sim}80%$ of total eggshell surface. Average coating films thickness for chitosan, starch, gelatin, dextrin and mineral oil were 2.2, 2.5, 3.9, 3.6 and $5.0\;{\mu}m$, respectively. After immersion process eggshell surface was almost completely covered by coating films. Chitosan coating was most effective among 5 film coatings in inhibiting growth of Salmonella enteritidis. Penetration process of Salmonella enteritidis through eggshell was investigated by multicolor imaging using CSLM and plate counting. Cuticle layer was the most important structure in blocking the penetration. Among 5 film coatings, chitosan showed the best and similar effectiveness with cuticle layer.

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Assessment of Inactivation for Salmonella spp. on Chicken Meat using Confocal Laser Microscopy and Flow Cytometry (공초점 현미경 및 유세포 분류기를 이용한 계육에서의 Salmonella균 불활성화 평가)

  • Jang, Keum-Il;Chung, Duck-Hwa;Ha, Sang-Do;Kim, Keun-Sung;Lee, Kyu-Ho;Kim, Min-Gon;Kim, Cheorl-Ho;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.38 no.2
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    • pp.290-294
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    • 2006
  • Inactivation rates of Salmonella enteritidis in vitro and in vivo were assessed using confocal microscopy and flow cytometry. S. enteritidis was inactivated with 1% (w/v) trisodium phosphate (TSP) and live cells, and inactive cells were distinguished by staining with fluorescent probe, LIVE/DEAD BacLight Bacteria Viability stain. After TSP treatment for 1 min, most of Salmonella cells changed from green (live cells) fluorescence to red (inactive cells) fluorescence, indication of effective sanitizing. Inactivation efficiency and contamination sites of S. enteritidis on chicken skin by TSP treatment were assessed using confocal laser microscopy. Precise flow cytometry histograms for viability changes of S. enteritidis. after TSP treatments were obtained. Efficiency of various sanitizer treatments on foodborne pathogens could be assessed using this method.

Morphological Analyses on Retinal Glial Responses to Glaucomatous Injury Evoked by Venous Cauterization

  • Lee, Ji-Yeon;Shin, Ji-Man;Chun, Myung-Hoon;Oh, Su-Ja
    • Applied Microscopy
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    • v.44 no.1
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    • pp.21-29
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    • 2014
  • Retinal glial responses to hypertensive glaucomatous injury were spatiotemporally surveyed. Retinas as a whole or vertical sections were processed for anti-glial fibrillary acidic protein (GFAP), anti-Iba1, anti-nerve growth factor (NGF), and anti-tumor necrosis factor (TNF)-${\alpha}$ immunohistochemistry for confocal microscopic analyses. The optic nerve head of paired controls was processed for electron microscopy. GFAP positive astrocytes appeared in the nerve fiber layer in the glaucomatous and control retinas, changing from fine protoplasmic to stout fibrous parallel to glaucomatous duration. Iba1 positive microglia appeared in both retinas, and enormous reaction appeared at the latest glaucomatous. M$\ddot{u}$ller reaction detected by GFAP reactivity expanded from the end feet to whole profile following to duration in the glaucomatous. NGF reactivity expended from the end feet to the proximal radial processes of the M$\ddot{u}$ller cells in both retinas according to glaucomatous duration. TNF-${\alpha}$ immunoreactivity in the nerve fiber layer was stronger in both the glaucomatous and controls than in the normal, and exceptionally at the latest glaucomatous was even lower than the normal. The astrocytes in the optic nerve head are interconnected with each other via gap junction. These results demonstrate that astrocyte reaction propagates to the contralateral via physical links, and TNF-${\alpha}$ is correlated with NGF production for neuroprotection in response to hypertensive glaucomatous injury.

Super-resolution Microscopy with Adaptive Optics for Volumetric Imaging

  • Park, Sangjun;Min, Cheol Hong;Han, Seokyoung;Choi, Eunjin;Cho, Kyung-Ok;Jang, Hyun-Jong;Kim, Moonseok
    • Current Optics and Photonics
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    • v.6 no.6
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    • pp.550-564
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    • 2022
  • Optical microscopy is a useful tool for study in the biological sciences. With an optical microscope, we can observe the micro world of life such as tissues, cells, and proteins. A fluorescent dye or a fluorescent protein provides an opportunity to mark a specific target in the crowd of biological samples, so that an image of a specific target can be observed by an optical microscope. The optical microscope, however, is constrained in resolution due to diffraction limit. Super-resolution microscopy made a breakthrough with this diffraction limit. Using a super-resolution microscope, many biomolecules are observed beyond the diffraction limit in cells. In the case of volumetric imaging, the super-resolution techniques are only applied to a limited area due to long imaging time, multiple scattering of photons, and sample-induced aberration in deep tissue. In this article, we review recent advances in super-resolution microscopy for volumetric imaging. The super-resolution techniques have been integrated with various modalities, such as a line-scan confocal microscope, a spinning disk confocal microscope, a light sheet microscope, and point spread function engineering. Super-resolution microscopy combined with adaptive optics by compensating for wave distortions is a promising method for deep tissue imaging and biomedical applications.

Quantitative Analysis of Pulp fiber Characteristics that Affect Paper Properties (II) (종이의 물성에 영향하는 섬유특성의 정량적 해석(II))

  • 이강진;박종문
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.32 no.2
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    • pp.35-39
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    • 2000
  • Refining is very important process of fibers treatment for proper paper properties. An extent of refining is usually measured by freeness, although freeness gives complicated meanings. One of a direct way of studying the refining effects on pulp fibers is making photomicrographs of beaten fibers. The conventional microscopy like light microscopy(LM) and scanning electron microscopy(SEM) require to preserve the wet structure of pulp fibers morphologically since most of papermaking process is carried out almost entirely in water. Recently developed microscopy, especially confocal laser scanning microscopy(CLSM), offers the possibility of examining fully hydrated pulp fibers. Cross-sectional images of wet pulp fibers are also generated using optical sectioning by CLSM and image analysis in order to verify and quantify the extent of fiber wall swelling indicating the internal fibrillation. At low beating load such as 2.5 kgf, in the same freeness, breaking length is higher than that of high beating load such as 5.6 kgf. fiber wall thickness at low beating load is greater than that at high beating load. This result is accounted for the fact that internal fibrillation in the low beating load was high.

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The study of property on the michelson interferometric microscopy (마이켈슨 간섭계형 현미경의 특성 연구)

  • 김경신;권남익
    • Korean Journal of Optics and Photonics
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    • v.10 no.5
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    • pp.369-372
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    • 1999
  • We have constructed an interferometric microscopy using a Michelson interferometer and a He-Ne laser. The three dimensional image was obtained by the interference from the reflected signal by a sample surface and from the reflected signal by a mirror. The axial resolution obtained by Michelson interferometric microscopy is as good as that of the white-light interferometer, but the same fringe is obtained when optical path difference is half-wavelength. The image from Michelson interferometric microscopy was compared with the images from the various types of confocal microscopy.

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