• 제목/요약/키워드: confocal microscopy

검색결과 437건 처리시간 0.033초

LIF 및 CLSM을 결합한 미소 간극 내 유체의 단면 온도 분포 측정 기법 (Measurement of Cross-sectional Temperature Distribution in Micro-scale Gap Fluid Using LIF Technique in Combination with CLSM)

  • 정동운;이상용
    • 대한기계학회논문집B
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    • 제30권9호
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    • pp.834-841
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    • 2006
  • In the present wort the Laser-induced Fluorescence (LIF) technique and Confocal Laser Scanning Microscopy (CLSM) have been combined to measure the temperature distribution across a micro-scale liquid layer as a direct and non-invasive method. Only the fluorescent light emitted from a very thin volume around a focal plane can be selectively detected, and it enables us to measure the liquid temperatures even at the close vicinity of the walls. As an experimental verification, a test section consists of two flat plates (for heating and cooling, respectively) separated by about 240 microns was made, and the methanol mixed with a temperature-sensitive dye, Rhodamine B, was filled in the gap between them. The measured temperature distribution across the gap showed good linearity, which is a typical characteristic of conduction heat transfer through a thin liquid layer. In result, the CLSM-LIF technique proposed in the present study was found to be a promising method to measure the local temperatures in the liquid flow field in microfluidic devices.

Bacterial Traits Involved in Colonization of Arabidopsis thaliana Roots by Bacillus amyloliquefaciens FZB42

  • Dietel, Kristin;Beator, Barbara;Budiharjo, Anto;Fan, Ben;Borriss, Rainer
    • The Plant Pathology Journal
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    • 제29권1호
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    • pp.59-66
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    • 2013
  • Colonization studies previously performed with a green-fluorescent-protein, GFP, labeled derivative of Bacillus amyloliquefaciens FZB42 revealed that the bacterium behaved different in colonizing surfaces of plant roots of different species (Fan et al., 2012). In order to extend these studies and to elucidate which genes are crucial for root colonization, we applied targeted mutant strains to Arabidopsis seedlings. The fates of root colonization in mutant strains impaired in synthesis of alternative sigma factors, non-ribosomal synthesis of lipopeptides and polyketides, biofilm formation, swarming motility, and plant growth promoting activity were analyzed by confocal laser scanning microscopy. Whilst the wild-type strain heavily colonized surfaces of root tips and lateral roots, the mutant strains were impaired in their ability to colonize root tips and most of them were unable to colonize lateral roots. Ability to colonize plant roots is not only dependent on the ability to form biofilms or swarming motility. Six mutants, deficient in abrB-, sigH-, sigD-, nrfA-, yusV and RBAM017410, but not affected in biofilm formation, displayed significantly reduced root colonization. The nrfA- and yusV-mutant strains colonized border cells and, partly, root surfaces but did not colonize root tips or lateral roots.

Modulation of Subcellular Ca2+ Signal by Fluid Pressure in Rat Atrial Myocytes

  • Woo Sun-Hee;Morad Martin
    • Biomolecules & Therapeutics
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    • 제14권1호
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    • pp.19-24
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    • 2006
  • Atrial chambers serve as mechanosensory systems during the haemodynamic or mechanical disturbances, which initiates arrhythmia. Atrial myocytes, lacking t-tubules, have two functionally separate sarcoplasmic reticulums (SRs): those at the periphery close to the surface membrane, and those at the cell interior (center) not associated with the membrane. To explore possible role of fluid pressure (FP) in the regulation of atrial local $Ca^{2+}$ signaling we investigated the effect of FP on subcellular $Ca^{2+}$ signals in isolated rat atrial myocytes using confocal microscopy. FP was applied to whole area of single myocyte with pressurized automatic micro-jet (200-400 $mmH_2O$) positioned close to the cell. Application of FP enhanced spontaneous occurrences of peripheral and central $Ca^{2+}$ sparks with larger effects on the peripheral release sites. Unitary properties of single sparks were not altered by FP. Exposure to higher FP often triggered longitudinal $Ca^{2+}$ wave. These results suggest that fluid pressure may directly alter excitability of atrial myocytes by activating $Ca^{2+}$-dependent ionic conductance in the peripheral membrane and by enhancing spontaneous activation of central myofilaments.

Anti-biofilm Activity of Xanthorrhizol Isolated from Curcuma xanthorrhiza Roxb. against Bacterial Biofilms Formed by Saliva and Artificial Multi-species Oral Strains

  • Yanti, Yanti;Rukayadi, Yaya;Lee, Kwan-Hyoung;Han, Sung-Hwa;Hwang, Jae-Kwan
    • Food Science and Biotechnology
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    • 제18권2호
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    • pp.556-560
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    • 2009
  • Xanthorrhizol, a sesquiterpene isolated from Curcuma xanthorrhiza Roxb., was used to investigate its effect on reducing the saliva and multi-species oral biofilms consisting of Streptococcus mutans, Streptococcus sanguis, and Actinomyces viscosus by anti-biofilm and confocal laser scanning microscopy (CLSM) assays. Xanthorrhizol exhibited significant antibiofilm activity in the dose- and time-dependent manners. Exposure to 2 and $5{\mu}g/mL$ xanthorrhizol for 30 min remained <50% of saliva and multi-species biofilms formed for 24 hr. In addition, exposure to $10{\mu}g/mL$ xanthorrhizol for 30 min reduced 65 and 77% of 24 hr saliva and multi-species oral biofilms, respectively. CLSM results visually demonstrated that xanthorrhizol reduced bacterial viability in the saliva and multi-species oral biofilms. These results suggest that C. xanthorrhiza Roxb. containing xanthorrhizol with strong anti-biofilm activity can be employed as a plant source for oral care functional foods.

Galactosylated Chitosan (GC)-graft-Poly(vinyl pyrrolidone) (PVP) as Hepatocyte-Targeting DNA Carrier: In Vitro Transfection

  • Park, In-Kyu;Jiang, Hu-Lin;Cook, Seung-Eun;Cho, Myung-Haing;Kim, Su-Il;Jeong, Hwan-Jeong;Akaike, Toshihiro;Cho , Chong-Su
    • Archives of Pharmacal Research
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    • 제27권12호
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    • pp.1284-1289
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    • 2004
  • Galactosylated chitosan-graft-poly(vinyl pyrrolidone) (GCPVP) was synthesized and characterized for hepatocyte-targeting gene carrier. GCPVP itself as well as GCPVP/DNA complex had negligible cytotoxicity regardless of the concentration of GCPVP and the charge ratio, but GCPVP/DNA complex had slightly cytotoxic effect on HepG2 cells only in the case of the higher charge ratio and 20 mM of $Ca^{2+}$ concentration used. Through the confocal laser scanning microscopy, it is shown that the endocytosis by interaction between galactose ligands of GCPVP and ASGPR of the hepatocytes was the major route of transfection of GCPVP/F-plasmid complexes.

Multi-dimensional analyses of plant chromosomes and genomes.

  • Fukui, Kiichi;Ohmido, Nobuko;Wako, Toshiyuki
    • 한국식물학회:학술대회논문집
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    • 한국식물학회 1998년도 The 12th Symposium on Plant Biotechnology Vol.12
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    • pp.61-70
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    • 1998
  • Genome and chromosome analyses in plants using fluorescence in situ hybridization (FISH) and immuno-staining (IMS) methods are reviewed by presenting the recent results obtained by the Chromosome Link, a group of chromosome and genome researchers. FISH is now effective to detect unique nucleotide sequences with 153 bp on the extended DNA fibers. Genomic in situ hybridization (GISH) also allows painting plant chromosomes of different genomes. GISH is quite effective to detect the genomic differentiation in the individual chromosomes within a nucleus. Three dimensional (3D) analyses are now available by confocal microscopy and a deconvolution system. These techniques are invaluable to visualize both the structural and functional dynamics within a nucleus. 3D-FISH revealed the spatial differentiation of different genomees within a nucleus. 3D-FISH also proved structural partition of centromeric and telomeric domains within a barely nucleus. The dynamic acetylation of histone H4 at the specific regions of a genome during a cell cycle is also analyzed using 3D-IMS. It is anticipated that these methods will provide us powerful tools to understand the structural and functional significance of plant chromosomes and genomes.

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Synthesis of a Novel Near-Infrared Fluorescent Dye: Applications for Fluorescence Imaging in Living Cells and Animals

  • Chen, Tongbin;Lai, Yijun;Huang, Suisheng
    • Bulletin of the Korean Chemical Society
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    • 제34권10호
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    • pp.2937-2941
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    • 2013
  • Fluorescence imaging is considered as one of the most powerful techniques for monitoring biomolecule activities in living systems. Near-infrared (NIR) light is advantageous for minimum photodamage, deep tissue penetration, and minimum background autofluorescence interference. Herein, we have developed a new NIR fluorescent dye, namely, RB-1, based on the Rhodamine B scaffold. RB-1 exhibits excellent photophysical properties including large absorption extinction coefficients, high fluorescence quantum yields, and high photostability. In particular, RB-1 displays both absorption and emission in the NIR region of the "biological window" (650-900 nm) for imaging in biological samples. RB-1 shows absorption maximum at 614 nm (500-725 nm) and emission maximum at 712 nm (650-825 nm) in ethanol, which is superior to those of traditional rhodamine B in the selected spectral region. Furthermore, applications of RB-1 for fluorescence imaging in living cells and small animals were investigated using confocal fluorescence microscopy and in vivo imaging system with a high signal-to-noise ratio (SNR = 10.1).

Effective Thermal Inactivation of the Spores of Bacillus cereus Biofilms Using Microwave

  • Park, Hyong Seok;Yang, Jungwoo;Choi, Hee Jung;Kim, Kyoung Heon
    • Journal of Microbiology and Biotechnology
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    • 제27권7호
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    • pp.1209-1215
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    • 2017
  • Microwave sterilization was performed to inactivate the spores of biofilms of Bacillus cereus involved in foodborne illness. The sterilization conditions, such as the amount of water and the operating temperature and treatment time, were optimized using statistical analysis based on 15 runs of experimental results designed by the Box-Behnken method. Statistical analysis showed that the optimal conditions for the inactivation of B. cereus biofilms were 14 ml of water, $108^{\circ}C$ of temperature, and 15 min of treatment time. Interestingly, response surface plots showed that the amount of water is the most important factor for microwave sterilization under the present conditions. Complete inactivation by microwaves was achieved in 5 min, and the inactivation efficiency by microwave was obviously higher than that by conventional steam autoclave. Finally, confocal laser scanning microscopy images showed that the principal effect of microwave treatment was cell membrane disruption. Thus, this study can contribute to the development of a process to control food-associated pathogens.

Expression of CD320 in human B cells in addition to follicular dendritic cells

  • Cho, Wha-Jung;Choi, Jin-Suk;Park, Chan-Hum;Yoon, Sun-Ok;Jeoung, Doo-Il;Kim, Young-Myeong;Choe, Jong-Seon
    • BMB Reports
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    • 제41권12호
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    • pp.863-867
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    • 2008
  • CD320 has been recently discovered and reported as a follicular dendritic cell (FDC) protein. Although CD320 is known to enhance proliferation of germinal center (GC) B cells, little other information is available. In this study, we investigated its cellular distribution in the GC. Confocal microscopy of human tonsil sections revealed co-localization of CD320 with CD19 and CD38 but not with CD3 indicating that GC B cells expressed CD320 in addition to FDC. In purified GC B cells, CD320 expression was inhibited in the nucleus, membrane and cytoplasm. Reverse transcriptase-polymerase chain reaction confirmed CD320 mRNA expression in B cells. These finding indicate that CD320 is expressed in B cells in addition to FDC, and that its GC activity may be more complicated than previously thought.

다양한 구조를 가진 알긴산-피브로인 비드 제조 (Preparation of Alginate-fibroin Beads with Diverse Structures)

  • 이진실;이신영;허원
    • KSBB Journal
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    • 제26권5호
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    • pp.422-426
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    • 2011
  • Alginate bead has been supplemented with various polymers to control permeability and to enhance mechanical strength. In this report, fibroin-reinforced alginate hydrogel was prepared, in which spatial localization of fibroin molecules was investigated. Confocal laser scanning microscopy revealed that fibroin molecules formed a fibrous network in the alginate-fibroin beads, which was expected to enhance mechanical strength as same as in many composite materials. Uniaxial compression test showed that fibroin-reinforced alginate beads had increased mechanical strength only after methanol treatment that caused ${\beta}$-sheet formation among fibroin molecules. Simultaneous curing and dialysis of alginate beads were carried out to remove excesscalcium but to retain fibroin in the dialysis chamber, which fabricated beads without internal fibrous fluorescent stains. Fibroin molecules were only found beneath the surface of the beads. The fibroin-diffused shell was further processed to form a thick wall after drying or was mobilizedto the centre of the bead by methanol treatment. Accordingly, the structure analyses provide processing methods of fibroin to form a wall or center clumps, which could be applied to design controlled delivery device.