• 제목/요약/키워드: fluorescence microscopy

검색결과 433건 처리시간 0.031초

Simple Analysis for Interaction between Nanoparticles and Fluorescence Vesicle as a Biomimetic Cell for Toxicological Studies

  • Umh, Ha Nee;Kim, Younghun
    • Bulletin of the Korean Chemical Society
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    • 제33권12호
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    • pp.3998-4002
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    • 2012
  • With continuing progress of nanotechnologies and various applications of nanoparticles, one needs to develop a quick and fairly standard assessment tool to evaluate cytotoxicity of nanoparticles. However, much cytotoxicity studies on the interpretation of the interaction between nanoparticles and cells are non-mechanistic and time-consuming. Here, we propose a simple screening method for the analysis of the interaction between several AgNPs (5.3 to 64 nm) and fluorescence-dye containing vesicles ($12{\mu}m$) acting as a biomimetic cell-membrane. Fluorescence-dye containing vesicle was prepared using a fluorescence probe (1,6-diphenyl-1,3,5-hexatryene), which was intercalated into the lipid bilayer due to their hydrophobicity. Zeta potential of all materials except for bare-AgNPs (+32.8 mV) was negative (-26 to -54 mV). The morphological change (i.e., rupture and fusion of vesicle, and release of dye) after mixing of the vesicle and AgNPs was observed by fluorescence microscopy, and fluorescence image were different with coating materials and surface charge of x-AgNPs. In the results, we found that the surface charge of nanoparticles is the key factor for vesicle rupture and fusion. This proposed method might be useful for analyzing the cytotoxicity of nanoparticles with cell-membranes instead of in vitro or in vivo cytotoxicity tests.

생쥐배의 생존성 평가에 있어 FDA의 이용 (The Use of FDA to Assess the Viability of Preimplantation Mouse Embryo In vitro)

  • 김재명;홍진기;서병희;이재현;정길생
    • Clinical and Experimental Reproductive Medicine
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    • 제19권1호
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    • pp.1-8
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    • 1992
  • A fluorescence microscopy technique using flurescein diacetateCFDA) as a substract has been tested for the evaluation of the viability of early mouse embryos. Embryos were incubated in T6 containing FDA concentrations of 2.5 to $50{\mu}g/ml$ for 1 to 5min. Embryos were then examined by reflected light fluorescence using a KP 490 and 520 barrier filter in a Nicon Diaphot microscopy. The results were as follow. 1. The rate of fluorescein accumulation increased on the concentration on FDA from $2.5{\times}10^{-6}M$ to $20{\times}10^{-6}M$ 2. The rate at which intracellular fluorescein was lost from embryos was depended on the temperature at which are stored. 3. Embryos with 3 min exposure to FDA have the most intensity of fluorescence. 4. Exposure of 2 cell embryos to FDA ($2.5-5{\mu}g/ml$) for 1 min did not alter their ability to delope normally in vitro.

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Dynamic lipopolysaccharide transfer cascade to TLR4/MD2 complex via LBP and CD14

  • Kim, Soo Jin;Kim, Ho Min
    • BMB Reports
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    • 제50권2호
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    • pp.55-57
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    • 2017
  • Toll-like receptor 4 (TLR4) together with MD2, one of the key pattern recognition receptors for a pathogen-associated molecular pattern, activates innate immunity by recognizing lipopolysaccharide (LPS) of Gram-negative bacteria. Although LBP and CD14 catalyze LPS transfer to the TLR4/MD2 complex, the detail mechanisms underlying this dynamic LPS transfer remain elusive. Using negative-stain electron microscopy, we visualized the dynamic intermediate complexes during LPS transfer-LBP/LPS micelles and ternary CD14/LBP/LPS micelle complexes. We also reconstituted the entire cascade of LPS transfer to TLR4/MD2 in a total internal reflection fluorescence (TIRF) microscope for a single molecule fluorescence analysis. These analyses reveal longitudinal LBP binding to the surface of LPS micelles and multi-round binding/unbinding of CD14 to single LBP/LPS micelles via key charged residues on LBP and CD14. Finally, we reveal that a single LPS molecule bound to CD14 is transferred to TLR4/MD2 in a TLR4-dependent manner. These discoveries, which clarify the molecular mechanism of dynamic LPS transfer to TLR4/MD2 via LBP and CD14, provide novel insights into the initiation of innate immune responses.

Lab-on-a-Chip for Monitoring the Quality of Raw Milk

  • Choi Jeong-Woo;Kim Young-Kee;Kim Hee-Joo;Lee Woo-Chang;Seong Gi-Hun
    • Journal of Microbiology and Biotechnology
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    • 제16권8호
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    • pp.1229-1235
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    • 2006
  • A lab-on-a-chip (LoC) was designed for simultaneous monitoring of microorganisms, antibiotic residues, somatic cells, and pH in raw milk. The LoC was fabricated from polydimethylsiloxane (PDMS) using microelectromechanical system (MEMS) technology, which consisted of two parts; a protein array and microchannel. The protein array was fabricated by immobilizing five types of antibodies corresponding to two microorganisms, two antibiotic residues, and somatic cells. A sol-gel film was deposited on a glass substrate to immobilize the antibodies. The target analytes in raw milk could be bound with the corresponding antibody by an immunoreaction, and the antigen-antibody complex was detected using fluorescence microscopy. SNARF-dextran was used as a pH indicator, and the SNARF-entrapped hydrogel was attached to the microchannel in the chip. After injecting the milk sample into the channel, the pH was measured by monitoring the change in fluorescence intensity by fluorescence microscopy. The on-chip simultaneous assay of two microorganisms (E. coli O157:H7 and Streptococcus agalactiae), two antibiotic residues (penicillin G and dihydrostreptomycin), and neutrophils was successfully accomplished using the proposed LoC system.

Comparative Study for Grinding of Two Cement Clinkers

  • Ibrahimi, Soumaya;Jamaa, Nejib Ben;Mliki, Khaoula;Bagane, Mohamed
    • International Journal of Concrete Structures and Materials
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    • 제5권2호
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    • pp.113-117
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    • 2011
  • The purpose of this work is the comparative study for grinding of two cement clinkers. X-ray fluorescence, physical and granulometric tests and optical microscopy were used to characterize the clinkers. Also grinding tests were carried out for ten samples to determine the parameters influencing grindability of its clinkers. The results of calculation of the energies of grinding according to the law developed by Von Rittinger and the study of the microstructure of the two clinkers shows good agreements. Indeed, frequent clusters of belite which indicate a lack of uniformity and fineness have an effect on lowering the grindability. The obtained analyses and the results enabled us to interpret the granulometry and the microstructure of clinker to control quality and resistance.

Fluorescence Spectroscopic and Atomic Force Microscopic Studies on the Microstructure of Polyimide/Silica-Titania Ternary Hybrid Composites

  • Park, Hae-Dong;Ahn, Ki-Youl;Mohammad A. Wahab;Jo, Nam-Ju;Kim, Il;Kim, Gyuhyun;Lee, Won-Ki;Ha, Chang-Sik
    • Macromolecular Research
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    • 제11권3호
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    • pp.172-177
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    • 2003
  • Biphenyltetracarboximide-phenylene diamine polyimide/silica-titania ternary hybrid composites were Prepared by thermal imidization and sol-gel reaction. Fluorescence spectroscopic, scanning electron microscopy, and atomic force microscopy studies revealed that the addition of small amount of titania showed profound effects on the microstructure and photophysical behaviors of polyimide/silica hybrid composites, when the content of silica-titania mixture was small or when the ratio of silica to titania in the mixture was high.

Potassium Distribution in the Apical Region of Rice Root

  • Rehman Shafiq;Park Moung-Ryoul;Ashraf Muhammad;Yun Song-Joong
    • 한국작물학회지
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    • 제51권4호
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    • pp.295-297
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    • 2006
  • Potassium (K) distribution in rice (Oryza sativa L.) root was studied by confocal laser microscopy, using potassium sensitive fluorescent dye potassium-binding benzofuran isophthalate (PBFI). Significantly high intensity of K-specific fluorescence was detected at the root cap region followed by meristematic and basal regions. A negligible or fainted fluorescence was observed at the root hairs area. These results suggest that K is heavily distributed in the apical area of rice root, which may be required in higher concentration for division and extension of cells, as it is the rapidly growing region of the root, moreover, may also be involved in water uptake by creating osmotic gradient across membranes.

자외선 형광장치현미경으로 본 화분관생장과 석탄 및 aureomycin의 상호작용에 대하여 (On the synergistic action of calcium and antibiotics in pollen growth as observed with ultraviolet fluorescence microscopy)

  • 곽병환
    • Journal of Plant Biology
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    • 제10권3_4호
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    • pp.1-3
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    • 1967
  • Observations were made on Crinum and Catharanthus pollen growth in artificial media by an ultraviolet transmission fluorescence microscope showing synergistic effect on pollen growth with calcium (Ca) and aureomycin. Bright yellow fluorescence of aureomycin enabling to trace out at tissue or cellular level did reveal that the greater accumulation of fluorescence occurred in the pollen tube wall if Ca was supplemented to the media than when aureomycin alone was present. The promotive pollen growth the media of Ca alone was further enhanced by the addtion of aureomycin. It was assumed that the promoted pollen growth with aureomycin in the Ca media was probably brought about by a supporting role of aureomycin in the Ca action.

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Measurements of Temperature and Flow Fields with Sub-Millimeter Spatial Resolution Using Two-Color Laser Induced Fluorescence (LIF) and Micro-Particle Image Velocimetry (PIV)

  • Kim Hyun Jung
    • Journal of Mechanical Science and Technology
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    • 제19권2호
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    • pp.716-727
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    • 2005
  • Comprehensive measurements for velocity and temperature fields have been conducted. A Micro PIV 2-color LIF system have been setup to measure the buoyancy driven fields in a 1-mm heated channel with low Grashof-Prandtl numbers [$86]. Fluorescence microscopy is combined with an MPIV system to obtain enough intensity images and clear pictures from nano-scale fluorescence particles. The spatial resolution of the Micro PIV system is $75{\mu}m\;by\;67{\mu}m$ and error due to Brownian motion is estimated $1.05\%$. Temperature measurements have achieved the $4.7\;{\mu}m$ spatial resolution with relatively large data uncertainties the present experiment. The measurement uncertainties have been decreased down to less than ${\pm}1.0^{\circ}C$ when measurement resolution is equivalent to $76\;{\mu}m$. Measured velocity and temperature fields will be compared with numerical results to examine the feasibility of development as a diagnostic technique.

High-speed Two-photon Laser Scanning Microscopy Imaging of in vivo Blood Cells in Rapid Circulation at Velocities of Up to 1.2 Millimeters per Second

  • Boutilier, Richard M.;Park, Jae Sung;Lee, Ho
    • Current Optics and Photonics
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    • 제2권6호
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    • pp.595-605
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    • 2018
  • The two-photon process of microscopy provides good spatial resolution and optical sectioning ability when observing quasi-static endogenous fluorescent tissue within an in vivo animal model skin. In order to extend the use of such systems, we developed a two-photon laser scanning microscopy system capable of also capturing $512{\times}512$ pixel images at 90 frames per second. This was made possible by incorporating a 72 facet polygon mirror which was mounted on a 55 kRPM motor to enhance the fast-scan axis speed in the horizontal direction. Using the enhanced temporal resolution of our high-speed two-photon laser scanning microscope, we show that rapid processes, such as fluorescently labeled erythrocytes moving in mouse blood flow at up to 1.2 mm/s, can be achieved.