• Title/Summary/Keyword: Fluorescent microscopy

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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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In Situ Fluorescence Optical Detection Using a Digital Micromirror Device (DMD) for 3D Cell-based Assays

  • Choi, Jong-Ryul;Kim, Kyujung;Kim, Donghyun
    • Journal of the Optical Society of Korea
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    • 제16권1호
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    • pp.42-46
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    • 2012
  • We have developed a fluorescence optical detection system using a digital micromirror device (DMD) for monitoring 3D cell culture matrices in situ. Full 3D imaging with fast scanning speed was implemented by the combined action of a DMD and a motorized stage. Imaging results with fluorescent microbeads measure the minimum axial resolution of the system as $6.3{\mu}m$, while full 1-mm scanning through 3D alginate-based matrix was demonstrated. For cell imaging, improved images were obtained by removing background fluorescence although the scanning distance was reduced because of low intracellular fluorescence efficiency. The system is expected to be useful to study various dynamics and behaviors of 3-dimensionally cultured cells in microfluidic systems.

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.

결핵진단의 면역학적 및 분자생물학적 방법 (Diagnosis of Tuberculosis; Serodiagnosis and Molecular Biologic Approach)

  • 신완식
    • Tuberculosis and Respiratory Diseases
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    • 제39권1호
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    • pp.1-6
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    • 1992
  • The diagnosis of tuberculosis is usually established using staining and culturing techniques. Fluorescent stains have improved the sensitivity of direct microscopy. Improved culture media coupled with radiometric means of detecting early mycobacterial growth have shortened the time needed for cultural diagnosis. Rapid immunodiagnostic techniques based on the detection of mycobacterial antigen or of antibodies to theses antigens have not, however, come into widespread clinical use. The DNA or RNA hybridization tests with labeled specific probes which have been described so far are not sensitive enough to be used for clinical speicimens without prior culturing. The advent of the polymerase chain reaction (PCR) has opened new possibilities for diagnosis of microbial infections. This technique has already been applied to a number of microorganisms. In the field of mycobacteria the PCR has been used to identify and to detect DNAs extracted from various mycobacteria. However, despite the extraordinary enthusiasm surrounding this technique and the considerable investiment, PCR has not emerged from the developmental "trenches" in the passed several years. It may be a considerable lenth of time before clinical microbiology laboratories become PCR playgrounds because many details remain to be worked out.

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One-Pot Synthesis of CdSe Quantum Dots Using Selenium Dioxide as a Selenium Source in Aqueous Solution

  • Wang, Yilin;Yang, Hong;Xia, Zhenyi;Tong, Zhangfa;Zhou, Liya
    • Bulletin of the Korean Chemical Society
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    • 제32권7호
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    • pp.2316-2318
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    • 2011
  • A novel technology has been developed for the synthesis of thioglycolic acid (TGA)-capped CdSe quantum dots (QDs) in aqueous medium. The reaction was carried out in air atmosphere with one-pot by using $SeO_2$ to replace Se or $Na_2Se$. The technological parameters including refluxing time, pH values and molar ratios of selenium to cadmium had significant influence on the luminescence properties of CdSe QDs. Furthermore, the obtained QDs were characterized by fluorescent spectroscopy, X-ray powder diffraction (XRD) and transmission electron microscopy (TEM), respectively. The results demonstrated that the CdSe QDs were of zinc-blended crystal structure in a sphere-like shape.

Studies on Sclerotium Formation in Curvularia Species

  • Singh, U.P.;Singh, S.K.;Sugawara, Koya;Srivastava, J.S.;Sarma, B.K.;Prithiviraj, B.
    • Mycobiology
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    • 제29권3호
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    • pp.154-159
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    • 2001
  • Natural sclerotium formation in two species of Curvularia was observed. The sclerotia were spherical to elongated. The scanning electron microscopical observations of sclerotia revealed that the sclerotia were of two distinct layers of cells, outer loosely woven hyphae and inner contact layer of cells. Different lights, viz. red, blue, green, fluorescent or addition of culture filtrate of Sclerotium rolfsii in the medium did not affect sclerotium formation.

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폴리에틸렌 담체에 부착된 혐기성 생물막 부착 특성 (Characteristics of attach of biomass on PE substratum under anaerobic condition)

  • 이승란;김도한;나영수;이창한;박영식;윤태경;송승구
    • 한국환경과학회지
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    • 제11권4호
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    • pp.327-332
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    • 2002
  • Optical microscope, SEM (Scanning Electron Microscopy) and fluorescent microscope were used for qualitative and morphological studies of the attached biomass on PE (polyethylene) substratum under anaerobic condition. It was shown by the observation of optical microscope that the initial attachment of biomass began in crevices of the surface of PE. The shape and structure of the attached biofilm could be observed by SEM photographs, but species of bacteria were and methanogens were not classified. A large number of methanogenic bacteria were identified on the surface of PE substratum by fluorescence under 480nm of radiation. It was estimated that methanogenic bacteria was also related to initial attachment of biomass under anaerobic condition.

잎권세균에 대한 개선된 형광현미경 관찰법 (Improved Epifluorescence Microscopy for Observation of Phyllosphere Bacteria on Leaf Surfaces)

  • 정필문;신광수;이인수;박성주
    • 미생물학회지
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    • 제37권1호
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    • pp.61-65
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    • 2001
  • 식물잎권의 잎표면세균에 대한 직접관찰과 이들을 분리시킨 용액 내 세균에 대한 간접관찰을 위하여 형광염료인 4', 6-diamidino-2-phenylindole(DAPI)와 acridine orange(AO)로 염색한 후 형광현미경을 사용하였다. AO로 염색한 잎표면은 배경형광이 진한 주홍색 이나 적색을 띠어 잎표면세균의 관잘 및 계수가 극히 힘들었으나 DAPI로 염색된 잎권세균은 뚜렷한 형광 영상을 나타내었다. 반면 잎표면세균 분리용액의 여과지에 대한 형광염색 결과는 DAPI와 AO 모두가 관찰이 가능하였다. 다만 DAPI가 AO에 비하여 형광 영상이 좀 더 뚜렷하였고 AO 염색과정에서는 염색된 여과지에 대한 세척과정이 필수적이었다. 최적의 DAPI 염색법은 잎과 여과지 모두가 5 $\mu$g/ml의 농도에서 5분 동안 염색하는 것이었다. 잎권의 형광현미경 관찰에 있어서 핵심적인 요소는 잎에서 나오는 수분을 최소화하는 것으로서 염색된 잎 시료를 거름종이 위에 올려놓고 공기 중에서 말리는 대신 $70^{\circ}C$를 유지한 건조기에서 2분 동안 말린 다음 바로 현미경으로 관찰하는 것이 가장 종은 방법인 것으로 확인되었다. 이렇게 확립된 형광현미경 관찰법을 떡갈나무의 잎권세균을 성공적으로 관찰하고 계수할 수 있었다.

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Root Colonization and ISR-mediated Anthracnose Disease Control in Cucumber by Strain Enterobacter asburiae B1

  • Bharathkumar, S.;Park, Jin-Woo;Han, Ji-Hee;Park, Kyung-Seok
    • The Plant Pathology Journal
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    • 제25권4호
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    • pp.333-343
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    • 2009
  • Here, we show that an endophytic bacterial strain, Enterobacter asburiae B1 exhibits the ability to elicit ISR in cucumber, tobacco and Arabidopsis thaliana. This indicates that strain B1 has a widespread ability to elicit ISR on various host plants. In this study, E. asburiae strain B1 did not show antifungal activity against tested major fungal pathogens, Colletotrichum orbiculare, Botrytis cinerea, Phytophthora capsici, Rhizoctonia solani, and Fusarium oxysporum. Moreover, the siderophore production by E. asburiae strain B1 was observed under in vitro condition. In greenhouse experiments, the root treatment of strain B1 significantly reduced disease severity of cucumber anthracnose caused by fungal pathogen C. orbiculare compared to nontreated control plants. By root treatment of strain B1 more than 50% disease control against anthracnose on cucumber was observed in all greenhouse experiments. Simultaneously, under the greenhouse condition, the soil drench of strain B1 and a chemical inducer benzothiadiazole (BTH) to tobacco plants induced GUS activity which is linked with activation of PR promoter gene. Furthermore, in Arabidopsis thaliana plants the soil drench of strain B1 induced the defense gene expression of PR1 and PDF1.2 related to salicylic acid and jasmonic acid/ethylene signaling pathways, respectively. In this study, for the main focus on root colonization by strain B1 associated with defense responses, bacterial cells of strain B1 was tagged with the gfp gene encoding the green fluorescent protein in order to determine the colonization pattern of strain B1 in cucumber. The gfp-tagged B1 cells were found on root surface and internal colonization in root, stem, and leaf. In addition to this, the scanning electron microscopy observation showed that E. asburiae strain B1 was able to colonized cucumber root surface.

메추리의 발달 중 배자 생식선에서 분리한 원시생식세포의 이동능에 대한 연구 (Studies on the Migratory Ability of Primordial Germ Cells from Embryonic Gonads at Different Developmental Stages in Quail)

  • D. K. Kim;G. H. Song;J. N. Kim;D. K. Jeong;K. D. Kim
    • 한국가금학회지
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    • 제28권1호
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    • pp.69-76
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    • 2001
  • Avian primordial germ cells (PGCs) originate from the epiblast and appear in the germinal crescent. These PGCs enter the developing blood vessels during stage 10∼12 (H&H), circulate in the blood stream, migrate into the developing gonadal anlage and differentiate into germ cells. However, it is not clear until when the migratory ability of PGC is maintained. This study was conducted to examine whether migratory ability is present in PGCs from the gonad at later embryonic developmental stages. In the present study, gonads were dissected from 5-, 6- and 10-day old quail embryos and treated with trypsin-EDTA. Gonadal PGCs (gPGCs) were purified by Ficoll-density-gradient-centrifugation and labeled with PKH26 fluorescent dye. The PKH26-labeled gPGCs were microinjected into the blood vessel of the recipient quail embryo. Manipulated recipients were incubated for 3 days, embedded in paraffin and sdctioned. The foreign gPGCs were detected by fluorescent and confocal laser microscopy. As a result, quail gPGCs, from 10, 6 and 5 day old embryos could migrate through the recipient blood stream at early stage and settle in the gonads. Thus, results suggest that gPGCs from upto 10-day old embryos keep properties seen in circulating PGC. Therefore, the PGCs of 10-day old embryonic gonads can be used for the tools of genetic manipulation.

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