• Title/Summary/Keyword: Phase contrast microscopy

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Estimation of Nanomechanical Properties of Nanosurfaces Using Phase Contrast Imaging in Atomic Force Microscopy (원자력현미경의 위상차영상을 이용한 나노표면의 미소기계적 특성 평가)

  • Ahn, Hyo-Sok
    • Transactions of the Korean Society of Machine Tool Engineers
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    • v.16 no.5
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    • pp.115-121
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    • 2007
  • Phase contrast imaging in atomic force microscopy showed a promise as an effective tool for better understanding of micromechanical properties of surfaces at nano scale. A qualitative estimation model for phase contrast images obtained with a tapping mode AFM was developed. This investigation demonstrated the high efficiency of combined analysis of topography and phase contrast images for characterizing nanosurfaces. Phase contrast images allowed estimation of relative stiffness(elastic modulus) of the sample surface. The phase contrast images revealed a significant inhomogeneity of the nano scale worn surfaces. Phase contrast images are also capable of revealing the formation of tribofilms.

Improving Phase Contrast of Digital Holographic Microscope using Spatial Light Modulator

  • Le, Thanh Bang;Piao, Meilan;Jeong, Jong-Rae;Jeon, Seok-Hee;Kim, Nam
    • Journal of the Optical Society of Korea
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    • v.19 no.1
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    • pp.22-28
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    • 2015
  • We propose a new method for improving the phase contrast of a multiphase digital holographic microscope using a spatial light modulator (SLM). Using the SLM as the annulus, our method improves the light contrast of the object edge to achieve higher accuracy. We demonstrate a digital holographic microscopy technique that provides a 30% improvement in the phase contrast compared to conventional microscopy, which utilizes a mechanical annulus. The phase-contrast improvement allows the 3D reconstructed hologram to be determined more precisely.

An Investigation of Worn DLC Coatings Using Atomic Force Microscopy (DLC코팅 마모면에 대한 원자력 현미경을 이용한 고찰)

  • ;;S. A. Chizhik
    • Tribology and Lubricants
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    • v.18 no.2
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    • pp.138-143
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    • 2002
  • Abstract - Tribofilms formed on won surface protect the DLC coating surface and decrease the fiction coefficient. However it is very difficult to evaluate their micromechanical properties due to their small thickness, inhomogeneity and discontinuity. The phase contrast images in tapping mode atomic farce microscopy allow an estimation of inhomogeneity in micromechanical properties of the sample surface. The purpose of this investigation is to demonstrate how the phase contrast images contribute to the characterization of thin tribofilms.

An investigation of worn DLC coatings using atomic force microscopy (DLC 코팅 마모면에 대한 원자력 현미경을 이용한 고찰)

  • ;;S.A.Chizhik
    • Proceedings of the Korean Society of Tribologists and Lubrication Engineers Conference
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    • 2001.06a
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    • pp.299-304
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    • 2001
  • Tribofilms formed on worn surface protect the DLC coating surface and decrease the friction coefficient. However it is very difficult to evaluate their micromechanical properties due to their small thickness, inhomogeneity and discontinuity. The phase contrast images in tapping mode atomic force microscopy allow an estimation of inhomogeneity in micromechanical properties of the sample surface. The purpose of this investigation is to demonstrate how the phase contrast images contribute to the characterization of thin tribofilms.

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Classification of Tumor cells in Phase-contrast Microscopy Image using Fourier Descriptor (위상차 현미경 영상 내 푸리에 묘사자를 이용한 암세포 형태별 분류)

  • Kang, Mi-Sun;Lee, Jeong-Eom;Kim, Hye-Ryun;Kim, Myoung-Hee
    • Journal of Biomedical Engineering Research
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    • v.33 no.4
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    • pp.169-176
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    • 2012
  • Tumor cell morphology is closely related to its migratory behaviors. An active tumor cell has a highly irregular shape, whereas a spherical cell is inactive. Thus, quantitative analysis of cell features is crucial to determine tumor malignancy or to test the efficacy of anticancer treatment. We use 3D time-lapse phase-contrast microscopy to analyze single cell morphology because it enables to observe long-term activity of living cells without photobleaching and phototoxicity, which is common in other fluorescence-labeled microscopy. Despite this advantage, there are image-level drawbacks to phase-contrast microscopy, such as local light effect and contrast interference ring. Therefore, we first corrected for non-uniform illumination artifacts and then we use intensity distribution information to detect cell boundary. In phase contrast microscopy image, cell is normally appeared as dark region surrounded by bright halo ring. Due to halo artifact is minimal around the cell body and has non-symmetric diffusion pattern, we calculate cross sectional plane which intersects center of each cell and orthogonal to first principal axis. Then, we extract dark cell region by analyzing intensity profile curve considering local bright peak as halo area. Finally, we calculated the Fourier descriptor that morphological characteristics of cell to classify tumor cells into active and inactive groups. We validated classification accuracy by comparing our findings with manually obtained results.

Nonparaxial Imaging Theory for Differential Phase Contrast Imaging

  • Jeongmin Kim
    • Current Optics and Photonics
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    • v.7 no.5
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    • pp.537-544
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    • 2023
  • Differential phase contrast (DPC) microscopy, a central quantitative phase imaging (QPI) technique in cell biology, facilitates label-free, real-time monitoring of intrinsic optical phase variations in biological samples. The existing DPC imaging theory, while important for QPI, is grounded in paraxial diffraction theory. However, this theory lacks accuracy when applied to high numerical aperture (NA) systems that are vital for high-resolution cellular studies. To tackle this limitation, we have, for the first time, formulated a nonparaxial DPC imaging equation with a transmission cross-coefficient (TCC) for high NA DPC microscopy. Our theoretical framework incorporates the apodization of the high NA objective lens, nonparaxial light propagation, and the angular distribution of source intensity or detector sensitivity. Thus, our TCC model deviates significantly from traditional paraxial TCCs, influenced by both NA and the angular variation of illumination or detection. Our nonparaxial imaging theory could enhance phase retrieval accuracy in QPI based on high NA DPC imaging.

Theory of Imaging And Diffraction (TEM 관련 이론해설 (3): 영상 형성이론과 회절이론)

  • Lee, Hwack-Joo
    • Applied Microscopy
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    • v.33 no.3
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    • pp.169-178
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    • 2003
  • In this review, theoretical approaches of imaging and diffraction in electron microscopy are introduced which allows the diffraction patterns and images to be treated with equal facility and emphasized the relationships between them. The coherent wave optics, incoherent wave imaging theory were introduced. The idea of Abbe theory was also introduced. Varoius phase contrast theories in small angle approximation were derived including the wave theory on Multi-component system.

Differential Interference Contrast Microscopic Module Using a Polarization Grating for Quantitative Phase Imaging (편광 격자 기반 정량적 위상 이미징을 위한 미분 간섭 현미경 모듈 개발)

  • Jin Hee Cho;Ki-Nam Joo
    • Korean Journal of Optics and Photonics
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    • v.34 no.6
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    • pp.261-268
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    • 2023
  • We propose a compact differential interference contrast microscopic module, which enables snapshot measurements for quantitative phase imaging. The proposed module adopts the lateral shearing interferometric principle, which can obtain self-interference without a reference. Due to the absence of the reference, the system is more stable than the typical interferometric systems. It uses a polarization grating to generate two laterally shifted wavefronts based on its birefringence and polarizing beam-splitting characteristics. Furthermore, the use of a polarization camera does not require sequential measurements for the phase extraction. In the experiments, we observe and measure the timely varying changes of various specimens to verify the system performance with the bright field images and phase contrast images. Because the proposed microscopic module also has the merit of being adaptable to typical microscopy instead of using an imaging camera, it can conveniently replace conventional contrast microscopy.

Consideration of Analysis Method of Asbestos and Existing Condition for Fibrous Materials in Domestic Air (국내 공기 중 섬유상 물질의 존재실태와 석면의 분석방법에 대한 고찰)

  • Hwang, Jinyeon;Oh, Jiho;Lee, Hyomin;Lee, Jinhyun;Son, Byeongseo
    • Journal of the Mineralogical Society of Korea
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    • v.28 no.2
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    • pp.165-171
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    • 2015
  • Dust samples collected from air in metropolitan areas of Busan and Ulsan were analyzed by phase contrast microscopy, phase contrast dispersion microscopy and SEM. Asbestos concentration in dust samples was lower than detection limit. Many fibrous materials were observed in dust samples, but most of them were organic fibers. Inorganic fibers such as rock fiber, ceramic fiber and gypsum were contained in the samples and non-fibrous minerals such as quartz, calcite and feldspar were also occasionally observed. Domestic law requires that asbestos in air dust is mainly analyzed by phase contrast microscopy. From this study, however, precise analysis of asbestos was almost impossible by this method only. As indicated in JIS method of Japan, therefore, count and identification of asbestos were more efficient by dispersion staining method after removing organic materials in samples by low temperature incinerator.

Applications of Digital Holography in Biomedical Microscopy

  • Kim, Myung-K.
    • Journal of the Optical Society of Korea
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    • v.14 no.2
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    • pp.77-89
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    • 2010
  • Digital holography (DH) is a potentially disruptive new technology for many areas of imaging science, especially in microscopy and metrology. DH offers a number of significant advantages such as the ability to acquire holograms rapidly, availability of complete amplitude and phase information of the optical field, and versatility of the interferometric and image processing techniques. This article provides a review of the digital holography, with an emphasis on its applications in biomedical microscopy. The quantitative phase microscopy by DH is described including some of the special techniques such as optical phase unwrapping and holography of total internal reflection. Tomographic imaging by digital interference holography (DIH) and related methods is described, as well as its applications in ophthalmic imaging and in biometry. Holographic manipulation and monitoring of cells and cellular components is another exciting new area of research. We discuss some of the current issues, trends, and potentials.