• Title/Summary/Keyword: confocal microscopy

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Specific Detection of DNA Using Quantum Dots and Magnetic Beads for Large Volume Samples

  • Kim, Yeon-Seok;Kim, Byoung-Chan;Lee, Jin-Hyung;Kim, Jung-Bae;Gu, Man-Bock
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.449-454
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    • 2006
  • Here we present a sensitive DNA detection protocol using quantum dots (QDs) and magnetic beads (MBs) for large volume samples. In this study, QDs, conjugated with streptavidin, were used to produce fluorescent signals while magnetic beads (MBs) were used to isolate and concentrate the signals. The presence of target DNAs leads to the sandwich hybridization between the functionalized QDs, the target DNAs and the MBs. In fact, the QDs-MBs complex, which is bound using the target DNA, can be isolated and then concentrated. The binding of the QDs to the surface of the MBs was confirmed by confocal microscopy and Cd elemental analysis. It was found that the fluorescent intensity was proportional to concentration of the target DNA, while the presence of non-complementary DNA produced no significant fluorescent signal. In addition, the presence of low copies of target DNAs such as 0.5 pM in large volume samples up to 40mL was successfully detected by using a magnet-assisted concentration protocol which consequently results in the enhancement of the sensitivity more than 100-fold.

A Study on the Anisotropic Flow Characteristics of Droplets on Rice Leaf Surface (벼 잎 표면에서 액적의 이방성 흐름 특성에 관한 연구)

  • Kim, Tae Wan
    • Tribology and Lubricants
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    • v.33 no.6
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    • pp.251-255
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    • 2017
  • In this study, we aimed to clarify the wettability and anisotropic flow characteristics of rice leaves as a basic study for engineering applications of anisotropic flow characteristics of rice leaf surface. To investigate the surface structure of rice leaf, the micro grooves and asperities of rice leaves were analyzed and quantified by scanning electron microscope, Confocal laser scanning microscopy, and stylus profilometer. The analysis of the structure of rice leaf surface confirmed that asymmetrical cone - like protrusions in leaf veins were inclined toward the leaf tip. The static contact angle test showed that the contact angle at the midline vein or leaf vein location where the micropapilla is concentrated is about $20^{\circ}$ higher than the leaf blade position. The contact angles of fresh and dried rice leave were also compared. The dried rice leaves showed a contact angle of about $5^{\circ}$ to $15^{\circ}$ higher than that of fresh leaves, suggesting that the volume of the protrusions decreased as the water was removed, thus reducing the contact area with the droplet. In the contact angle history test the hysteresis in the leaf tip direction was found to be much lower than that in the leaf petiole direction. This results can be explained that asymmetrical cone - like protrusions had a significant effect on the droplet flow characteristics through contact angle hysteresis experiment.

Use of Cationic PAM as a Surface Sizing Additive to Improve Paper Properties

  • Seo, Man-Seok;Lee, Hak-Lae;Youn, Hye-Jung
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2006.06b
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    • pp.245-250
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    • 2006
  • This study was focused on the use of cationic PAM (Polyacrylamide) as a surface sizing additive to improve the surface sizing properties of paper. Effects of the ionic property, viscosity and charge density of PAM on bending stiffness of surface sized papers were investigated. Use of cationic PAM as a surface sizing additive improved bending stiffness while addition of anionic PAM did not show any effect. Increase of starch holdout with the addition of cationic PAM was attributed as a prime reason of stiffness increase. Viscosity of PAM was one of the most important factors affecting surface sizing due to its influence on the interaction between cationic PAM and oxidized starch solution. Greater improvement of bending stiffness of paper was obtained when high charged PAM was used as an additive. The order of addition was found to have significant influence on the effect of additives since it influences the formation of network structure among starch, cationic PAM, and SA (styrene acrylic acid copolymer). Investigation on the penetration of starch solution was carried out with CLSM (Confocal Laser Scanning Microscopy), and it was shown that the addition of cationic PAM to oxidized starch solution made starch molecules stay on the paper surface rather than penetrating into the paper structure because of the electrostatic interaction between negatively charged fibers and positively charged cationic PAM.

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Plastid Transformation in the Monocotyledonous Cereal Crop, Rice (Oryza sativa) and Transmission of Transgenes to Their Progeny

  • Lee, Sa Mi;Kang, Kyungsu;Chung, Hyunsup;Yoo, Soon Hee;Ming Xu, Xiang;Lee, Seung-Bum;Cheong, Jong-Joo;Daniell, Henry;Kim, Minkyun
    • Molecules and Cells
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    • v.21 no.3
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    • pp.401-410
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    • 2006
  • The plastid transformation approach offers a number of unique advantages, including high-level transgene expression, multi-gene engineering, transgene containment, and a lack of gene silencing and position effects. The extension of plastid transformation technology to monocotyledonous cereal crops, including rice, bears great promise for the improvement of agronomic traits, and the efficient production of pharmaceutical or nutritional enhancement. Here, we report a promising step towards stable plastid transformation in rice. We produced fertile transplastomic rice plants and demonstrated transmission of the plastidexpressed green fluorescent protein (GFP) and aminoglycoside 3′-adenylyltransferase genes to the progeny of these plants. Transgenic chloroplasts were determined to have stably expressed the GFP, which was confirmed by both confocal microscopy and Western blot analyses. Although the produced rice plastid transformants were found to be heteroplastomic, and the transformation efficiency requires further improvement, this study has established a variety of parameters for the use of plastid transformation technology in cereal crops.

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

  • Jeong, Dong-Woon;Lee, Sang-Yong
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.30 no.9 s.252
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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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    • v.29 no.1
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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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    • v.14 no.1
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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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    • v.18 no.2
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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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    • v.27 no.12
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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
    • Proceedings of the Botanical Society of Korea Conference
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    • 1998.07a
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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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