• Title/Summary/Keyword: Quantitative phase microscopy

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DigitalMicrograph Script Source Listing for a Geometric Phase Analysis

  • Kim, Kyou-Hyun
    • Applied Microscopy
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    • 제45권2호
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    • pp.101-105
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    • 2015
  • Numerous digital image analysis techniques have been developed with regard to transmission electron microscopy (TEM) with the help of programming. DigitalMicrograph (DM, Gatan Inc., USA), which is installed on most TEMs as operational software, includes a script language to develop customized software for image analysis. Based on the DM script language, this work provides a script source listing for quantitative strain measurements based on a geometric phase analysis.

살아있는 세포 영상획득을 위한 common-path phase microscopy (Common-path phase microscopy for lives cell imaging)

  • 이지용;이승락;;김덕영
    • 한국광학회:학술대회논문집
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    • 한국광학회 2008년도 하계학술발표회 논문집
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    • pp.273-274
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    • 2008
  • We present a quantitative phase microscopy for live cells. This method uses the principles of common path inteferometry and single shot phase image. This system has the ability to measure live cells quantitatively with subnanometer path length stability and millisecond scale aquisition time.

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Nonparaxial Imaging Theory for Differential Phase Contrast Imaging

  • Jeongmin Kim
    • Current Optics and Photonics
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    • 제7권5호
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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.

Applications of Digital Holography in Biomedical Microscopy

  • Kim, Myung-K.
    • Journal of the Optical Society of Korea
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    • 제14권2호
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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.

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

  • 조진희;주기남
    • 한국광학회지
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    • 제34권6호
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    • pp.261-268
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    • 2023
  • 본 연구에서는 간단한 구조의 층밀림 간섭계 기반의 정량적 위상 이미징을 위한 미분 간섭 현미경 모듈을 제안한다. 제안하는 모듈은 복굴절성과 편광 광 분할기의 특성을 갖는 기하 위상 소자 중의 하나인 편광 격자를 이용하여 층밀림 간섭계 구조로 구성되고, 편광 카메라를 통해 획득된 4개의 편광 영상으로 정량적인 위상차 영상뿐만 아니라 일반적인 명시야 현미경 영상을 동시에 추출할 수 있다. 또한, 제안하는 모듈의 실시간 영상 획득 가능성, 안정성 및 소형화로 인해 기존의 현미경에 편리하게 적용할 수 있다. 제안하는 시스템을 검증하기 위해 다양한 시편에 대한 명시야 영상 및 위상차 영상을 획득하였고, 또한 이들 영상들을 합성하여 보다 가독성이 높은 시편 영상을 획득하였다.

SOFC 음극용 Ni-YSZ 복합체의 미세구조와 전기적 물성간의 상관관계 : I. 미세구조 분석 (Correlatin between the Microstructure and the Electrical Conductivity of SOFC anode, Ni-YSZ : I. Microstructure Analysis)

  • 문환;이해원;이종호;윤기현
    • 한국세라믹학회지
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    • 제37권5호
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    • pp.479-490
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    • 2000
  • The microstructure of Ni-YSZ composite as an anode of SOFC was investigated as a function of Ni content(10-70 vol%) in order to examine the correlation between microstructural-and electrical property. Image analysis based on quantitative microscopy theory was performed to quantify the microstructural property. We could get the informations about the size and distribution, contiguity and interfacial area of each phase or between the phases from the image analysis. According to the image analysis, contiguity between the same phae was mainly dependent on the amount of the phase while the contiguity between different phases was additionally influenced by the microstructural changes, especailly by the coarsening of the Ni phase. The whole length of pores perimeter was increased as Ni content increased, which indicated the overall microstructural evolution was mostly related with the coarsening of Ni phase. Ni-Ni interfacial area was also gradually increased as Ni content increased but controlled by pore phase at low Ni content region and by YSZ phase at intermediate Ni content region. These quantified microstructural properties were used to characterize the electrical properties of Ni-YSZ composite.

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

  • 강미선;이정엄;김혜련;김명희
    • 대한의용생체공학회:의공학회지
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    • 제33권4호
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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.

Statistical Analysis of 3D Volume of Red Blood Cells with Different Shapes via Digital Holographic Microscopy

  • Yi, Faliu;Lee, Chung-Ghiu;Moon, In-Kyu
    • Journal of the Optical Society of Korea
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    • 제16권2호
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    • pp.115-120
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    • 2012
  • In this paper, we present a method to automatically quantify the three-dimensional (3D) volume of red blood cells (RBCs) using off-axis digital holographic microscopy. The RBCs digital holograms are recorded via a CCD camera using an off-axis interferometry setup. The RBCs' phase image is reconstructed from the recorded off-axis digital hologram by a computational reconstruction algorithm. The watershed segmentation algorithm is applied to the reconstructed phase image to remove background parts and obtain clear targets in the phase image with many single RBCs. After segmenting the reconstructed RBCs' phase image, all single RBCs are extracted, and the 3D volume of each single RBC is then measured with the surface area and the phase values of the corresponding RBC. In order to demonstrate the feasibility of the proposed method to automatically calculate the 3D volume of RBC, two typical shapes of RBCs, i.e., stomatocyte/discocyte, are tested via experiments. Statistical distributions of 3D volume for each class of RBC are generated by using our algorithm. Statistical hypothesis testing is conducted to investigate the difference between the statistical distributions for the two typical shapes of RBCs. Our experimental results illustrate that our study opens the possibility of automated quantitative analysis of 3D volume in various types of RBCs.

Single Shot White Light Interference Microscopy for 3D Surface Profilometry Using Single Chip Color Camera

  • Srivastava, Vishal;Inam, Mohammad;Kumar, Ranjeet;Mehta, Dalip Singh
    • Journal of the Optical Society of Korea
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    • 제20권6호
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    • pp.784-793
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    • 2016
  • We present a single shot low coherence white light Hilbert phase microscopy (WL-HPM) for quantitative phase imaging of Si optoelectronic devices, i.e., Si integrated circuits (Si-ICs) and Si solar cells. White light interferograms were recorded by a color CCD camera and the interferogram is decomposed into the three colors red, green and blue. Spatial carrier frequency of the WL interferogram was increased sufficiently by means of introducing a tilt in the interferometer. Hilbert transform fringe analysis was used to reconstruct the phase map for red, green and blue colors from the single interferogram. 3D step height map of Si-ICs and Si solar cells was reconstructed at multiple wavelengths from a single interferogram. Experimental results were compared with Atomic Force Microscopy and they were found to be close to each other. The present technique is non-contact, full-field and fast for the determination of surface roughness variation and morphological features of the objects at multiple wavelengths.

회절위상현미경을 이용한 광섬유의 굴절률 프로파일 측정 (Measurement of Refractive Index Profile of Optical Fiber Using the Diffraction Phase Microscope)

  • ;문석배
    • 한국광학회지
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    • 제23권4호
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    • pp.135-142
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    • 2012
  • 본 연구에서는 공동경로간섭계(common-path interferometer)에 기반한 회절위상현미경(diffraction phase microscopy)을 이용한 광섬유의 굴절률 프로파일(refractive index profile) 측정기술을 개발하였다. 투과형 회절격자를 이용하여 광섬유 시료를 통과한 빛으로부터 핀홀을 이용하여 영의 공간주파수 성분만을 갖는 기준광을 생성하고, 기준광을 다시 시료의 위상정보를 갖는 시료광과 간섭시키는 방법을 통해 시료의 위상정보를 가진 간섭무늬를 형성시켰다. 이렇게 얻어진 간섭 이미지로부터 수치적 처리과정을 거쳐 공간적 위상정보 곧, 위상 이미지를 획득하고 이 데이터를 역아벨변환(inverse Abel transform)을 통해 굴절률 프로파일로 변환할 수 있었다. 이때 클래딩과 광섬유 주변의 매질 사이의 굴절률차로 인해 발생하는 배경위상을 이론적으로 얻어진 함수형태에 맞춰 예측하고 이를 측정된 위상에서 제거하는 배경위상제거 방법을 개발하여 사용하였다. 이를 통해 광섬유 코어 부근의 위상정보만으로도 굴절률 프로파일을 성공적으로 이뤄질 수 있음이 입증되었다. 본 연구를 통하여 회절위상현미경 특유의 측정 안정성과 편의성을 가진 광섬유 굴절률 프로파일 측정장치를 개발하였고 광섬유 및 도파로의 굴절률 분포를 비파괴적으로 분석할 수 있어 광섬유 및 광섬유소자 개발에 활용될 수 있을 것으로 기대된다.