• Title/Summary/Keyword: Dark field microscopy

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Dark Field Digital Holographic Microscopy Based on Two-lens 360-degree Oblique Illumination

  • Zhang, Xiuying;Zhao, Yingchun;Yuan, Caojin;Feng, Shaotong;Wang, Lin
    • Current Optics and Photonics
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    • v.4 no.3
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    • pp.193-199
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    • 2020
  • In this paper we propose a dark-field digital holographic microscopy system based on 360-degree oblique illumination. This setup is constructed without using a dark-field objective. The principle of 360-degree oblique illumination of vortex beam and dark-field digital holographic microscopy are introduced theoretically and experimentally. By analyzing the reconstructed image of a dark-field digital hologram of a USAF 1951 target, it is proved that the imaging resolution can be improved by this method. And also, comparison and analysis are made on the reconstructed image of a bright-dark field digital hologram of a pumpkin stem slice, the result shows that the imaging contrast is also enhanced with this method, and it is effective for dark-field digital holographic microscopy imaging of large transparent biological samples.

Dark-field Transmission Electron Microscopy Imaging Technique to Visualize the Local Structure of Two-dimensional Material; Graphene

  • Na, Min Young;Lee, Seung-Mo;Kim, Do Hyang;Chang, Hye Jung
    • Applied Microscopy
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    • v.45 no.1
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    • pp.23-31
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    • 2015
  • Dark field (DF) transmission electron microscopy image has become a popular characterization method for two-dimensional material, graphene, since it can visualize grain structure and multilayer islands, and further provide structural information such as crystal orientation relations, defects, etc. unlike other imaging tools. Here we present microstructure of graphene, particularly, using DF imaging. High-angle grain boundary formation wass observed in heat-treated chemical vapor deposition-grown graphene on the Si substrate using patch-quilted DF imaging processing, which is supposed to occur by strain around multilayer islands. Upon the crystal orientation between layers the multilayer islands were categorized into the oriented one and the twisted one, and their local structure were compared. In addition information from each diffraction spot in selected area diffraction pattern was summarized.

Reflective Fourier Ptychographic Microscopy Using Segmented Mirrors and a Mask

  • Ahn, Hee Kyung;Chon, Byong Hyuk
    • Current Optics and Photonics
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    • v.5 no.1
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    • pp.40-44
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    • 2021
  • In this paper, LED arrays with segmented mirrors and a mask are presented as a new dark-field illuminator for reflective Fourier ptychographic microscopy (FPM). The illuminator can overcome the limitations of the size and the position of samples that the dark-field illuminator using a parabolic mirror has had. The new concept was demonstrated by measuring a USAF 1951 target, and it resolved a pattern in group 10 element 6 (274 nm) in the USAF target. The new design of the dark-field illuminator can enhance competitiveness of the reflective FPM as a versatile measurement method in industry.

Image Processing of Defocus Series TEM Images for Extracting Reliable Phase Information (정확한 위상정보를 얻기 위한 탈초점 영상들의 이미지 처리기법)

  • Song, Kyung;Shin, Ga-Young;Kim, Jong-Kyu;Oh, Sang-Ho
    • Applied Microscopy
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    • v.41 no.3
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    • pp.215-222
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    • 2011
  • We discuss the experimental procedure for extracting reliable phase information from a defocus series of transmission electron microscopy (TEM) dark-field images using the transport of intensity equation (TIE). Taking InGaN/GaN multi-quantum well light-emitting diode as a model system, various factors affecting the final result of reconstructed phase such as TEM sample preparation, TEM imaging condition, image alignment, the correction of defocus values and the use of high frequency pass filter are evaluated. The obtained phase of wave function was converted to the geometric phase of the corresponding lattice planes, which was then used for the two-dimensional mapping of lattice strain following the dark-field inline holography (DIH) routine. The strain map obtained by DIH after optimized image processing is compared with that obtained by the geometric phase analysis of high resolution TEM (HRTEM) image, manifesting that DIH yields more accurate and reliable strain information than HRTEM-based GPA.

Development of Dark Field image Processing Technique for the Investigation of Nanostructures

  • Jeon, Jongchul;Kim, Kyou-Hyun
    • Journal of Powder Materials
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    • v.24 no.4
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    • pp.285-291
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    • 2017
  • We propose a custom analysis technique for the dark field (DF) image based on transmission electron microscopy (TEM). The custom analysis technique is developed based on the $DigitalMicrograph^{(R)}$ (DM) script language embedded in the Gatan digital microscopy software, which is used as the operational software for most TEM instruments. The developed software automatically scans an electron beam across a TEM sample and records a series of electron diffraction patterns. The recorded electron diffraction patterns provide DF and ADF images based on digital image processing. An experimental electron diffraction pattern is recorded from a IrMn polycrystal consisting of fine nanograins in order to test the proposed software. We demonstrate that the developed image processing technique well resolves nanograins of ~ 5 nm in diameter.

Simple Analysis for Interaction between Nanoparticles and Dye-Containing Vesicles as a Biomimetic Cell-Membrane

  • Shin, Sohyang;Umh, Ha Nee;Kim, Younghun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.1
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    • pp.231-236
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    • 2013
  • Some cytotoxicity studies for the interpretation of the interaction between nanoparticles and cells are non-mechanistic and time-consuming. Therefore, non-biological screening methods, which are faster and simpler than in-vivo and in-vitro methods, are required as alternatives to current cytotoxicity tests. Here, we proposed a simple screening method for the analysis of the interaction between several AgNPs (bare-, citrate-, and polyvinylpyrrolidone-coating) and dye-containing vesicles acting as a biomimetic cell-membrane. The interaction between AgNPs and vesicles could be evaluated readily by UV-vis spectra. Absorbance deviation in UV-vis spectra revealed a large attraction between neighboring particles and vesicles. This was confirmed by (Derjagin, Landau, Verwey, and Overbeek) theory and DMF (dark-field microscopy) analysis. This proposed method might be useful for analyzing the cytotoxicity of nanoparticles with cell-membranes instead of in vitro or in vivo cytotoxicity tests.

Real-time Monitoring of Colloidal Nanoparticles using Light Sheet Dark-field Microscopy Combined with Microfluidic Concentration Gradient Generator (μFCGG-LSDFM)

  • Choe, Hyeokmin;Nho, Hyun Woo;Park, Jonghoon;Kim, Jin Bae;Yoon, Tae Hyun
    • Bulletin of the Korean Chemical Society
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    • v.35 no.2
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    • pp.365-370
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    • 2014
  • For real-time monitoring of colloidal nanoparticles (NPs) in aqueous media, a light sheet type dark-field microscopy system combined with a microfluidic concentration gradient generator (${\mu}FCGG$-LSDFM) was developed. Various concentrations of colloidal Au NPs were simultaneously generated with the iFCGG and characterized with the LSDFM setup. The number concentrations and hydrodynamic size distributions were measured via particle counting and tracking analysis (PCA and PTA, respectively) approaches. For the 30 nm Au NPs used in this study, the lower detection limit of the LSDFM setup was 3.6 ng/mL, which is about 400 times better than that of optical density measurements under the same ${\mu}FCGG$ system. Additionally, the hydrodynamic diameter distribution of Au NPs was estimated as $39.7{\pm}12.2nm$ with the PTA approach, which agrees well with DLS measurement as well as the manufacturer's specification. We propose this ${\mu}FCGG$-LSDFM setup with features of automatic generation of NP concentration gradient and real-time monitoring of their physicochemical characteristics (e.g., number concentration, and hydrodynamic size distribution) as an important component of future high-throughput screening or high-content analysis platforms of nanotoxicity.

Detection of Leptospires in Experimentally Infected Mice, Using Fluorescent Antibody Technique (형광항체법(螢光抗體法)을 이용(利用)한 실험적(實驗的) 감염(感染) 마우스에서의 Leptospira균(菌)의 검출(檢出))

  • Seuk, H.B.;Seo, I.S.
    • Korean Journal of Veterinary Research
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    • v.13 no.1
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    • pp.39-46
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    • 1973
  • Cultural method, dark field microscopy & fluorescent antibody technique were compared for their sensitivity of the detection of leptospires from experimentally infected mice. Two groups of mice were infected with L. icterohemorrhagiae (M20) and L. australis (Ballico), and the infected blood, urine and a number of organs were subjected to the bacterial isolation. The results obtained were summarized as follows: 1. L. icterohemorrhagiae (M20) and L. australis (Ballico) in blood, urine and various tissues of experimentally infected mice were detected with a negrigible non specificity, by the fluorescent antibody technique. 2. The fluorescent antibody technique, as applied to detection of leptospires in blood, urine and various infected tissue, proved to be better than cultural method and dark-field microscopy. 3. Early detection of leptospires by fluorescent antibody technique were possible in blood at 2 days after inoculation, whereas detection of organisms in liver, spleen, lung and kidney were observed later. By means of fluorescent antibody technique, the detection of leptospires in kidney and urine was possible up to 34 days postinoculation, whereas those in other parts were impossible. 4. Fluorescent antibody reaction of leptospires were highly specific to homologous antigen rather than to heterologous one. 5. Fluorescent antibody technique may be of value in the application for the demonstration of leptospira from clinical specimens.

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Detail analysis of the peak disappearance of minor phase in mechanically alloyed samples(II) (기계적 합금화 시료에서 미소상 피이크의 소멸현상 해석(II))

  • Kim, Hye-Sung
    • Journal of the Korean Society of Industry Convergence
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    • v.4 no.1
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    • pp.27-34
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    • 2001
  • Refining of powder particles and their dissolution into the Al matrix during mechanical alloying(MA) were investigated by using X-ray diffraction(XRD) transmission electron microscopy (TEM) functions of alloy composition, milling time and ball to powder ratio(BPR). It is found that Ti particles less than 20nm are observed in a dark field image of mechanically alloyed Al-10wt%Ti whose XHD pattern exhibits no Ti peak. The observed change of lattice constant of AI indicates that about 1 wt%Ti can he solved in Al after MA for a long time, independent of alloy composition, milling time and BPR, suggesting that most of Ti particles arc retained in the Al matrix. It is concluded that the disappearance of XRD peaks in mechanically alloyed Al-10wt%Ti is not simply attributable to the dissolution of Ti into Al, but associated mainly with extreme refining and/or heavy straining of Ti Particles.

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Sequential conversion from line defects to atomic clusters in monolayer WS2

  • Gyeong Hee Ryu;Ren-Jie Chan
    • Applied Microscopy
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    • v.50
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    • pp.27.1-27.6
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    • 2020
  • Transition metal dichalcogenides (TMD), which is composed of a transition metal atom and chalcogen ion atoms, usually form vacancies based on the knock-on threshold of each atom. In particular, when electron beam is irradiated on a monolayer TMD such as MoS2 and WS2, S vacancies are formed preferentially, and they are aligned linearly to constitute line defects. And then, a hole is formed at the point where the successively formed line defects collide, and metal clusters are also formed at the edge of the hole. This study reports a process in which the line defects formed in a monolayer WS2 sheet expends into holes. Here, the process in which the W cluster, which always occurs at the edge of the formed hole, goes through a uniform intermediate phase is explained based on the line defects and the formation behavior of the hole. Further investigation confirms the atomic structure of the intermediate phase using annular dark field scanning transition electron microscopy (ADF-STEM) and image simulation.