• Title/Summary/Keyword: fluorescence microscopy.

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Confocal laser scanning microscopy image를 이용한 UASB granule의 메탄 생성 능력 측정

  • Lee, Yu-Jin;Kim, Hyo-Seop;An, Yeong-Hui;Park, Seong-Hun
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.365-369
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    • 2000
  • Methanogenic activity of granular sludge was monitored by specific methanogenic activity (SMA) assay and confocal laser scanning microscopy (CLSM) during start-up of a thermophilic UASB reactor. Autofluorescence by CLSM could visualize the methanogenic bacterial population inside sludge granules and its intensity was proportional to SMA. Considering the complex procedures of SMA measurement, fluorescence quantification by CLSM can be suggested as a routine technique measuring methanogenic activity in UASB granules.

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Ultrathin Polymer Networks of Itaconic Acid Copolymers and Poly(allkylamine) by the ;angmuir-Blodgett Technique

  • 최기선;이범종;장상목;권영수
    • Bulletin of the Korean Chemical Society
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    • v.16 no.6
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    • pp.493-498
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    • 1995
  • Ultrathin networks of itaconic acid copolymers and poly(allylamine) were produced by a Langmuir-Blodgett (LB) technique employing a double-chain amine as a monolayer template which was subsequently removed by extraction after thermal crosslinking. Itaconic acid copolymers used were copoly (itaconic acid-ethyl vinyl ether) and copoly (itaconic acid-n-butyl vinyl ether). The polyion-complexed monolayers of three components consisting of template amine, itaconic acid copolymer and poly (allylamine) were formed at the air-water interface. The Langmuir film properties have been studied by the surface pressure-area isotherm and fluorescence microscopy. The monolayers were transferred on solid substrates and were characterized by FT-IR spectroscopy, X-ray photoelectron spectroscopy, and scanning electron microscopy (SEM). Two-dimensional polymer networks were formed through imide or amide linkages by heat treatment under vacuum. The heat-treated films were extracted with chloroform after immersion in aq. sodium chloride to remove the template amines. SEM observation of a LB film on a porous fluorocarbon membrane filter with pore diameter of 0.1 μm showed covering of the pores by six layers in the polyion complex state.

Changes of Cytosolic $Ca^{2+}$ under Metabolic Inhibition in Isolated Rat Ventricular Myocytes

  • Kang, Sung-Hyun;Kim, Na-Ri;Joo, Hyun;Youm, Jae-Boum;Park, Won-Sun;Warda, Mohamed;Kim, Hyung-Kyu;Von Cuong, Dang;Kim, Tae-Ho;Kim, Eui-Yong;Han, Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.5
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    • pp.291-298
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    • 2005
  • To characterize cytosolic $Ca^{2+}$ fluctuations under metabolic inhibition, rat ventricular myocytes were exposed to $200{\mu}M$ 2,4-dinitrophenol (DNP), and mitochondrial $Ca^{2+}$, mitochondrial membrane potential (${\Delta}{\Psi}m$), and cytosolic $Ca^{2+}$ were measured, using Rhod-2 AM, TMRE, and Fluo-4 AM fluorescent dyes, respectively, by Laser Scanning Confocal Microscopy (LSCM). Furthermore, the role of sarcolemmal $Na^+$/$Ca^{2+}$ exchange (NCX) in cytosolic $Ca^{2+}$ efflux was studied in KB-R7943 and $Na^+$-free normal Tyrode's solution (143 mM LiCl ). When DNP was applied to cells loaded with Fluo-4 AM, Fluo-4 AM fluorescence intensity initially increased by $70{\pm}10$% within $70{\pm}10$ s, and later by $400{\pm}200$% at $850{\pm}45$ s. Fluorescence intensity of both Rhod-2 AM and TMRE were initially decreased by DNP, coincident with the initial increase of Fluo-4 AM fluorescence intensity. When sarcoplasmic reticulum (SR) $Ca^{2+}$ was depleted by $1{\mu}M thapsigargin plus $10{\mu}M ryanodine, the initial increase of Fluo-4 AM fluorescence intensity was unaffected, however, the subsequent progressive increase was abolished. KB-R7943 delayed both the first and the second phases of cytosolic $Ca^{2+}$ overload, while $Na^+$-free solution accelerated the second. The above results suggest that: 1) the initial rise in cytosolic $Ca^{2+}$ under DNP results from mitochondrial depolarization; 2) the secondary increase is caused by progressive $Ca^{2+}$ release from SR; 3) NCX plays an important role in transient cytosolic $Ca^{2+}$ shifts under metabolic inhibition with DNP.

Initial bacterial adhesion on resin, titanium and zirconia in vitro

  • Lee, Byung-Chul;Jung, Gil-Yong;Kim, Dae-Joon;Han, Jung-Suk
    • The Journal of Advanced Prosthodontics
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    • v.3 no.2
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    • pp.81-84
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    • 2011
  • PURPOSE. The aim of this in vitro study was to investigate the adhesion of initial colonizer, Streptococcus sanguis, on resin, titanium and zirconia under the same surface polishing condition. MATERIALS AND METHODS. Specimens were prepared from Z-250, cp-Ti and 3Y-TZP and polished with $1 {\mu}m$ diamond paste. After coating with saliva, each specimen was incubated with Streptococcus sanguis. Scanning electron microscope, crystal violet staining and measurement of fluorescence intensity resulting from resazurin reduction were performed for quantifying the bacterial adhesion. RESULTS. Surface of resin composite was significantly rougher than that of titanium and zirconia, although all tested specimens are classified as smooth. The resin specimens showed lower value of contact angle compared with titanium and zirconia specimens, and had hydrophilic surfaces. The result of scanning electron microscopy demonstrated that bound bacteria were more abundant on resin in comparison with titanium and zirconia. When total biofilm mass determined by crystal violet, absorbance value of resin was significantly higher than that of titanium or zirconia. The result of relative fluorescence intensities also demonstrated that the highest fluorescence intensity was found on the surface of resin. Absorbance value and fluorescence intensity on titanium was not significantly different from those on zirconia. CONCLUSION. Resin specimens showed the roughest surface and have a significantly higher susceptibility to adhere Streptococcus sanguis than titanium and zirconia when surfaces of each specimen were polished under same condition. There was no significant difference in bacteria adhesion between titanium and zirconia in vitro.

Green Synthesis of Multifunctional Carbon Nanodots and Their Applications as a Smart Nanothermometer and Cr(VI) Ions Sensor

  • Li, Lu;Shao, Congying;Wu, Qian;Wang, Yunjian;Liu, Mingzhu
    • Nano
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    • v.13 no.12
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    • pp.1850147.1-1850147.14
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    • 2018
  • In this work, water-soluble and blue-emitting carbon nanodots (CDs) were synthesized from apple peels for the first time via one-step hydrothermal method. The synthetic route is facile, green, economical and viable. The as-prepared CDs were characterized thoroughly by transmission electron microscopy (TEM), X-ray diffraction (XRD), Raman, Fourier transform infrared (FT-IR), X-ray photoelectron (XPS), fluorescence and UV-Vis absorption spectroscopy in terms of their morphology, surface functional groups and optical properties. The results show that these CDs possessed ultrasmall size, good dispersivity, and high tolerance to pH, ionic strength and continuous UV irradiation. Significantly, the CDs had fast and reversible response towards temperature, and the accurate linear relationship between fluorescence intensity and temperature was used to design a novel nanothermometer in a broad temperature range from 5 to $65^{\circ}C$ facilely. In addition, the fluorescence intensity of CDs was observed to be quenched immediately by Cr(VI) ions based on the inner filter effect. A low-cost Cr(VI) ions sensor was proposed employing CDs as fluorescent probe, and it displayed a wide linear range from 0.5 to $200{\mu}M$ with a detection limit of $0.73{\mu}M$. The practicability of the developed Cr(VI) sensor for real water sample assay was also validated with satisfactory recoveries.

A Study on Match and Mismatch DNA Hybridization properties Using DNA Hybridization Detection Sensor (DNA Hybridization 검출 센서를 이용한 매치 및 미스매치 DNA hybridization 특성 연구)

  • Kim, Do-Kyun;Kwon, Young-Soo
    • Proceedings of the KIEE Conference
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    • 2003.10a
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    • pp.89-91
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    • 2003
  • The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and other application area. So, the improvement of DNA detection system is very important for the determination of this hybridization reaction. In this study, we report the characterization of the probe and target oligonucleotide hybridization reaction using the evanescent field microscopy. First, we have fabricated DNA chip microarray. The particles which were immobilized oligonucleotides were arranged by the random fluidic self-assembly on the pattern chips, using hydrophobic interaction. Second, we have detected DNA hybridization reaction using evanescent field microscopy. The 5'-biotinylated probe oligonucleotides were immobilized on the surface of DNA chip microarray and the hybridization reaction with the Rhodamine conjugated target oligonucleotide was excited fluorescence generated on the evanescent field microscopy. In the foundation of this result, we could be employed as the basis of a probe olidonucleotide, capable of detecting the target oligonucleotide and monitoring it in a large analyte concentration range and various mismatching condition.

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Bufalin Induces Mitochondrial Pathway-Mediated Apoptosis in Lung Adenocarcinoma Cells

  • Ding, Da-Wei;Zhang, Yong-Hong;Huang, Xin-En;An, Qing;Zhang, Xun
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.23
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    • pp.10495-10500
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    • 2015
  • Background: To evaluate the effects of bufalin in A549 human lung adenocarcinoma epithelial cells in vitro and assess the underlying mechanisms. Materials and Methods: Human A549 non-small cell lung cancer (NSCLC) cells were treated with various concentrations of bufalin. Cell proliferation was measured by CCK-8 assay, apoptotic cell percentage was calculated by flow cytometry and morphological change was observed by inverted phase contrast microscopy/transmission electron microscopy. In addition, the membrane potential of mitochondria was detected by JC-1 fluorescence microscopy assay, and the related protein expression of cytochrome C and caspase-3 was analyzed by Western blotting. Results: Bufalin could inhibit the proliferation of A549 cells via induction of apoptosis, with the evidence of characteristic morphological changes in the nucleus and mitochondria. Furthermore, bufalin decreased the mitochondrial membrane potential with up-regulation of cytochrome C in the cytosol, and activation of caspase-3. Conclusions: Bufalin inhibits the proliferation of A549 cells and triggers mitochondria-dependent apoptosis, pointing to therapeutic application for NSCLC.

Melatonin Protects Human Adipose-Derived Stem Cells from Oxidative Stress and Cell Death

  • Tan, Shaun S.;Han, Xiaolian;Sivakumaran, Priyadharshini;Lim, Shiang Y.;Morrison, Wayne A.
    • Archives of Plastic Surgery
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    • v.43 no.3
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    • pp.237-241
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    • 2016
  • Background Adipose-derived stem cells (ASCs) have applications in regenerative medicine based on their therapeutic potential to repair and regenerate diseased and damaged tissue. They are commonly subject to oxidative stress during harvest and transplantation, which has detrimental effects on their subsequent viability. By functioning as an antioxidant against free radicals, melatonin may exert cytoprotective effects on ASCs. Methods We cultured human ASCs in the presence of varying dosages of hydrogen peroxide and/or melatonin for a period of 3 hours. Cell viability and apoptosis were determined with propidium iodide and Hoechst 33342 staining under fluorescence microscopy. Results Hydrogen peroxide (1-2.5 mM) treatment resulted in an incremental increase in cell death. 2 mM hydrogen peroxide was thereafter selected as the dose for co-treatment with melatonin. Melatonin alone had no adverse effects on ASCs. Co-treatment of ASCs with melatonin in the presence of hydrogen peroxide protected ASCs from cell death in a dose-dependent manner, and afforded maximal protection at $100{\mu}M$ (n=4, one-way analysis of variance P<0.001). Melatonin co-treated ASCs displayed significantly fewer apoptotic cells, as demonstrated by condensed and fragmented nuclei under fluorescence microscopy. Conclusions Melatonin possesses cytoprotective properties against oxidative stress in human ASCs and might be a useful adjunct in fat grafting and cell-assisted lipotransfer.

Apoptosis Induction, Cell Cycle Arrest and in Vitro Anticancer Activity of Gonothalamin in a Cancer Cell Lines

  • Alabsi, Aied M.;Ali, Rola;Ali, Abdul Manaf;Al-Dubai, Sami Abdo Radman;Harun, Hazlan;Kasim, Noor H. Abu;Alsalahi, Abdulsamad
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.10
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    • pp.5131-5136
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    • 2012
  • Cancer is one of the major health problems worldwide and its current treatments have a number of undesired adverse side effects. Natural compounds may reduce these. Currently, a few plant products are being used to treat cancer. In this study, goniothalamin, a natural occurring styryl-lactone extracted from Goniothalamus macrophyllus, was investigated for cytotoxic properties against cervical cancer (HeLa), breast carcinoma (MCF-7) and colon cancer (HT29) cells as well as normal mouse fibroblast (3T3) using MTT assay. Fluorescence microscopy showed that GTN is able to induce apoptosis in HeLa cells in a time dependent manner. Flow cytometry further revealed HeLa cells treated with GTN to be arrested in the S phase. Phosphatidyl serine properties present during apoptosis enable early detection of the apoptosis in the cells. Using annexin V/PI double staining it could be shown that GTN induces early apoptosis on HeLa cells after 24, 48 and 72 h. It could be concluded that goniothalamin showing a promising cytotoxicity effect against several cancer cell lines including cervical cancer cells (HeLa) with apoptosis as the mode of cell death induced on HeLa cells by Goniothalamin was.

Image Stitching Using Normalized Cross-Correlation and the Thresholding Method in a Fluorescence Microscopy Image of Brain Tumor Cells (정규 상호상관도 및 이진화 기법을 이용한 뇌종양 세포의 형광 현미경 영상 스티칭)

  • Seo, Ji Hyun;Kang, Mi-Sun;Kim, Hyun-jung;Kim, Myoung-Hee
    • Journal of Korea Multimedia Society
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    • v.20 no.7
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    • pp.979-985
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    • 2017
  • This paper, which covers a fluorescence microscopy image of brain tumor cells, looks at drug reactions by treating different types and concentrations of drugs on a plate of $24{\times}16$ wells. Due to the limitation of the field of view, a well was taken into 9 field images, and each has an overlapping area with its neighboring fields. To analyze more precisely, image stitching is needed. The basic method is finding a similar area using normalized cross-correlation (NCC). The problem is that some overlapping areas may not have any duplicated cells that help to find the matching point. In addition, the cell objects have similar sizes and shapes, which makes distinguishing them difficult. To avoid calculating similarity between blank areas and roughly distinguishing different cells, thresholding is added. The thresholding method classifies background and cell objects based on fixed thresholds and finds the location of the first seen cell. After getting its location, NCC is used to find the best correlation point. The results are compared with a simple boundary stitched image. Our proposed method stitches images that are connected in a grid form without collision, selecting the best correlation point among areas that contain overlapping cells and ones without it.