• 제목/요약/키워드: fluorescence microscopy.

검색결과 435건 처리시간 0.029초

Spontaneous Nanoparticle Formation From a Fluorescent Nucleoside Analogue

  • Bang, Eun-Kyoung;Moon, Do-Hyun;Kim, Byeang-Hyean
    • Bulletin of the Korean Chemical Society
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    • 제32권spc8호
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    • pp.2906-2910
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    • 2011
  • A fluorescent nucleoside analogue, $^AC$, featuring two non-complementary nucleobases linked through an ethynyl group, was synthesized. The extended ${\pi}$-conjugation imparts $^AC$ with red-shifted absorbance (relative to adenine and cytosine) and pale-blue fluorescence. It spontaneously forms nanoparticles, which exhibit considerably enhanced fluorescence, without the help of any additional stabilizing agent. The DMSO/water ratio was an important factor influencing the construction of the NPs. X-ray crystallography confirmed the structure of $^AC$; dynamic light scattering and scanning electron microscopy confirmed the existence of the nanoparticles.

Association of Aster Yellow Phytoplasma with Witches′ Broom Disease of Ash(Fraxinus rhynchophylla Hence) in Korea

  • Sangsub Han;Lim, Tae-Heon;Byeongjin Cha
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.73.2-73
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    • 2003
  • Typical whiches broom symptoms caused by phytoplasma were observed in Ash (Fraxinus rhynchophylla Hence) in Korea. The symptoms were showing abnormally small leaves, short internodes, and proliferation of shoots. Fluorescence and electron microscopy of leaf midribs revealed phytoplasma positive DAPI fluorescence and numerous phytoplasma bodies localized in the phloem sieve tubes. Phytoplasma DNA of 1.8 Kb was detected consistently from all symptomatic samples by the amplification of phytoplasma DNA with the phytoplasma specific primer pair Pl/P7. But no phytoplasma DNA was detected in healthy ash seedlings. Based on sequence analyses of an amplified region, this phytoplasma is closely related to Eqilodium phyllody, Mulberry dwarf, and Aster yellows phytoplasmas with the homology of 99.95 %, 99.79 % and 99.78 %, respectively, This phylogenetic analyses indicate that ash witches broom phytoplasma but is evidently distinct from the ash yellows group 16SrⅦ and should be classified into the Aster yellows group 16SrⅥ.

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Fabrication of Protein A-Viologen Hetero Langmuir- Blodgett Film for Fluorescence Immunoassay

  • Lee, Woochang;Chun, Bum-Suk;Oh, Byung-Keun;Lee, Won-Hong;Park, Jeong-Woo
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제9권4호
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    • pp.241-244
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    • 2004
  • Protein A molecular thin film was fabricated as a platform of antibody-based biosensor. For the immobilization of the protein A thin film, a viologen multilayer was built up using the Langmuir-Blodgett (LB) technique, and then, protein A was adsorbed on the viologen LB film by an electrostatic interaction force, which was formed as a hetero-film structure. For the deposition of viologen, surface pressure area ($\pi$-A) isotherm was investigated. The fabricated protein A-viologen hetero LB film was investigated using atomic force microscopy (AFM). Using the developed molecular film, antibody immobilization and fluorescence measurement was carried out.

Comparative lectin binding patterns of Cochlodinium polykrikoides Margalef

  • Rhodes, Lesley L.;Cho, Yong-Chul;Cho, Eun-Seob
    • Journal of the korean society of oceanography
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    • 제35권3호
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    • pp.153-157
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    • 2000
  • Four different FITC-conjugated lectins were used to visually evaluate lectin binding activity by optical staining quality using confocal laser scanning microscopy (CLSM) of Cochzodinium polykrikoides in nature (wild type) and culture (cultured type). Cells from the field and cultures treated with ConA fluoresced only at the outer cell wall, and the abundance and distribution of the fluorescent signal were similar. Treatment with PWM and HPA did not elicit fluorescence at the cell surface, but the wild type exposed to HPA showed greater binding than did the cultured cells, possibly due to greater concentrations of glucosamine. The wild type cells treated with LBL lectin showed a strong green fluorescence on the cell surface, whereas cultured cells did not. Signal intensity and abundance were greater than for any other lectins tested in this study. These results suggest that wild type and cultured type are significantly different based on surface sugar production. In particular, the wild type cells apear richer in galactosamine-like moieties. Neither glucose nor mannose-like moieties were present in either wild types or cultured cells.

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Polarity Probing Two-Photon Fluorophores Based on [2.2]Paracyclophane

  • Woo, Han-Young;Korystov, Dmitry;Jin, Young-Eup;Suh, Hong-Suk
    • Bulletin of the Korean Chemical Society
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    • 제28권12호
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    • pp.2253-2260
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    • 2007
  • A series of tetra donor substituted [2.2]paracyclophane-based two-photon absorption (TPA) fluorophores were synthesized in neutral and cationic forms. The imaging activity of overall set of fluorophores was studied by the two-photon induced fluorescence (TPIF) method in a range of solvents. We also measured a clear progression toward a longer photoluminescence lifetime with increasing solvent polarity (intrinsic photoluminescence lifetime, τi: ~2 ns in toluene → 12-16 ns in water). The paracyclophane fluorophores with this unique property can be utilized as an optical polarity probe for the biomolecular substrates. The combined measurement of the two-photon fluorescence microscopy (TPM) cell image and TPIF lifetime can give us a better understanding of the biological processes and local environments in the cells.

Signal increasing method in confocal scanning microscopy in fluorescence mode using curved mirror

  • Kang, Dong-kyun;Seo, Jung-woo;Gweon, Dae-gab
    • 제어로봇시스템학회:학술대회논문집
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    • 제어로봇시스템학회 2001년도 ICCAS
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    • pp.99.3-99
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    • 2001
  • In fluorescence mode confocal scanning microscope, level of detected signal is very low. In object scanning type confocal scanning microscope, the additional optical system with objective lens and plane mirror was proposed to increase signal intensity, but there was none for beam scanning type confocal scanning microscope. We propose reflecting optical systems which improve signal intensity in beam scanning type confocal scanning microscope. We choose one of the proposed optical systems and design the optical system, i.e., select optical components and assign distances between the selected components. To design the optical system, we use finite ray tracing method and make cost function to be minimized.

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스마트폰 기반 Mobile SmartScope를 이용한 혈구 영상 분석 (Analysis of Blood Cell Images Using Smartphone-based Mobile SmartScope)

  • 박춘호;조명옥;이동희;김중경
    • 한국가시화정보학회지
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    • 제10권2호
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    • pp.25-31
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    • 2012
  • High-performance smartphones, equipped with a digital camera and an application software, can render conventional bench-top laboratory instruments mobile at affordable costs. As the smartphone-based devices are portable and wireless, they have wide applications especially in providing point-of-care (POC) tests in resource-constrained areas. We developed a hand-held diagnostic system, Mobile SmartScope, which consists of a small optical unit integrated with a smartphone. The performance of the SmartScope was favorably compared with that of conventional light microscopy in detecting and quantifying red blood cells. We also evaluated the fluorescence detection limit of the SmartScope incorporated with a blue light-emitting diode and appropriate optical filters by using fluorescently labeled microbeads for intensity calibration.

Caspase-11 Promoter-GFP Construct as a Dual Reporter of Cytotoxicity and Inflammation

  • Shin, Ki-Soon;Kang, Shin-Jung
    • Animal cells and systems
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    • 제10권2호
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    • pp.73-77
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    • 2006
  • Caspase-11 has been known as a dual regulator of apoptosis and inflammatory response. An unusual feature of caspase-11 is that its expression is induced by apoptotic or proinflammatory stimuli. Utilizing these unusual features of caspase-11, we have developed a simple and sensitive assay method to screen pro- or anti-apoptotic/inflammatory molecules. To develop this assay method, we generated a reporter construct where GFP expression is regulated by caspase-11 promoter. When several types of cultured cells were transfected with this reporter construct and subsequently treated with various apoptotic or proinflammatory molecules, expression of GFP by the activation of caspase-11 promoter was easily detected by fluorescence microscopy or spectrofluorometry. In addition, a reduction of the GFP fluorescence was detected when an agent reported to suppress caspase-11 induction was applied. These results suggest that our reporter system can be used to screen pro- or anti-apoptotic/inflammatory molecules.

Single-Molecule Methods for Investigating the Double-Stranded DNA Bendability

  • Yeou, Sanghun;Lee, Nam Ki
    • Molecules and Cells
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    • 제45권1호
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    • pp.33-40
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    • 2022
  • The various DNA-protein interactions associated with the expression of genetic information involve double-stranded DNA (dsDNA) bending. Due to the importance of the formation of the dsDNA bending structure, dsDNA bending properties have long been investigated in the biophysics field. Conventionally, DNA bendability is characterized by innate averaging data from bulk experiments. The advent of single-molecule methods, such as atomic force microscopy, optical and magnetic tweezers, tethered particle motion, and single-molecule fluorescence resonance energy transfer measurement, has provided valuable tools to investigate not only the static structures but also the dynamic properties of bent dsDNA. Here, we reviewed the single-molecule methods that have been used for investigating dsDNA bendability and new findings related to dsDNA bending. Single-molecule approaches are promising tools for revealing the unknown properties of dsDNA related to its bending, particularly in cells.

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.