• Title/Summary/Keyword: Fluorescent probe

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Measurement of Cell Death Constant in Anabaena flos-aquae (Cyanophyceae) by the Molecular Probe (Anabaena flos-aquae 에서의 세포사멸계수(Cell Death Constant)의 측정)

  • 오인혜
    • The Korean Journal of Ecology
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    • v.20 no.3
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    • pp.169-173
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    • 1997
  • The measurement of cell death constant in Anabaena flos-aquae was tested by the Live/Dead BacLight Viability kit(Molecular Probes Co., Seatle, WA). When the Live/Dead BacLight Viability kit was applied to Anabaena flos-aquae, the cells with intact cell membranes(live cells) stained fluorescent green, while the cell with damaged membranes(dead cells) stained fluorescent red and the background remained virtually nonfluorescent. The rations of live : dead cells in the cell suspension were controlled artifically and Live/Dead BacLight Viability kit was applied to them. The ratios of green:red fluorescent cells in the cell suspension were the same as those of live : dead cells controlled artifically. It was also approved by the fluorescence emission. The cell death constant was measured in the P-limited Anabaena flos-aquae chemostal culture in the N-fixing and $KNO_3-supplied$ conditions. The culture in N-fixing chemostat had a dead cell proportion of 1.2% at the growth rate of 0.7/day and increased to 2.6% at the growth rate of 0.3/day. The cell death constant of N-fixing culture was 0.008/day.There was a same trend in the $KNO_3-supplied$ chemostat culture. The proportion of dead cell was 1.6% of dead cell proportion at the growth rate of 0.7/day and increased to 4.3% at the growth rate of 0.3/day.

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Selective Fe2+ Ion Recognition Using a Fluorescent Pyridinyl-benzoimidazole-derived Ionophore

  • Lee, Jeong Ah;Eom, Geun Hee;Park, Hyun Min;Lee, Ju Hoon;Song, Hyesun;Hong, Chang Seop;Yoon, Sungho;Kim, Cheal
    • Bulletin of the Korean Chemical Society
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    • v.33 no.11
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    • pp.3625-3628
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    • 2012
  • Fluorescent organic molecules that respond to changes in the $Fe^{2+}$ concentration with selectivity to other abundant di-valent metal ions will offer the ability to understand the dynamic fluctuations of the $Fe^{2+}$ ion in interesting media. The use of 6-Br-ppmbi, derived from 2-pyridin-2-yl-benzimidazole, for metal ion-selective fluorescence recognition was investigated. Screening of the main group and transition metal ions showed exclusive selectivity for $Fe^{2+}$ ions even in the presence of competing metal ions. In addition, the requirement for low concentrations of probe molecules to detect certain amounts of $Fe^{2+}$ ions make this sensor unique compared to previously reported $Fe^{2+}$ ion sensors.

FMN-Based Fluorescent Proteins as Heavy Metal Sensors Against Mercury Ions

  • Ravikumar, Yuvaraj;Nadarajan, Saravanan Prabhu;Lee, Chong-Soon;Jung, Seunho;Bae, Dong-Ho;Yun, Hyungdon
    • Journal of Microbiology and Biotechnology
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    • v.26 no.3
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    • pp.530-539
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    • 2016
  • Bacterial light-oxygen-voltage-sensing photoreceptor-derived flavin mononucleotide (FMN)-based fluorescent proteins act as a promising distinct class of fluorescent proteins utilized for various biomedical and biotechnological applications. The key property of its independency towards oxygen for its chromophore maturation has greatly helped this protein to outperform the other fluorescent proteins such as GFP and DsRed for anaerobic applications. Here, we describe the feasibility of FMN-containing fluorescent protein FbFP as a metal-sensing probe by measuring the fluorescence emission changes of a protein with respect to the concentration of metal ions. In the present study, we demonstrated the mercury-sensing ability of FbFP protein and the possible amino acids responsible for metal binding. A ratiometric approach was employed here in order to exploit the fluorescence changes observed at two different emission maxima with respect to Hg2+ at micromolar concentration. The engineered variant FbFPC56I showed high sensitivity towards Hg2+ and followed a good linear relationship from 0.1 to 3 μM of Hg2+. Thus, further engineering with a rational approach would enable the FbFP to be developed as a novel and highly selective and sensitive biosensor for other toxic heavy metal ions as well.

The Early Detection of the Spore Using Sonication and Fluorescent Dye in the Field (현장에서 초음파 파쇄와 형광시약을 이용한 포자의 조기 탐지)

  • Ha, Yeon-Chul;Choi, Ki-Bong
    • KSBB Journal
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    • v.26 no.4
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    • pp.305-310
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    • 2011
  • This study was carried out to establish the optimum condition of cell disruption with a sonicator for the detection of the spore, Bacillus anthracis ${\Delta}$-sterne for the purpose of developing automatic fluorometer. The efficiency of sonication on the ${\Delta}$-sterne spore disruption was very weak. The ${\Delta}$-sterne spore with zirconia bead showed greater disruption than the ${\Delta}$-sterne spore alone when sonificated. The volumn of the zirconia bead added in the spore solution has little effect on the disruption efficiency. The detection limit of the ${\Delta}$-sterne spore with zirconia bead and the ${\Delta}$-sterne spore alone was $10^6$ CFU/mL and $5{\times}10^7$ CFU/mL respectively, when sample was sonicated for 20 seconds with a sonicator probe of 13 mm diameter.

Non-Invasive Environmental Detection using Heat Shock Gene-Green Fluorescent Protein Fusions

  • Cha, Hyeong-Jun
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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    • pp.355-356
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    • 2000
  • Three 'stress probe' plasmids were constructed and characterized which utilize a green fluorescent protein (CFP) as a non-invasive reporter to elucidate Escherichia coli cellular stress responses in quiescent or 'resting' cells. Facile detection of cellular stress levels was achieved by fusion of three heat shock stress protein promoter elements, those of the heat shock transcription factor ${\sigma}^{32}$, pretense subunit ClpB, and chaperone DnaK, to the reporter gene $gfp_{uv}$. When perturbed by chemical or physical stress (such as heat shock, nutrient (amino acid) limitation, addition of IPTG, acetic acid, ethanol, phenol, antifoam, and salt (osmotic shock), the E. coli cells produced GFPuv which was easily detected from within the cells as emitted green fluorescence. A temporal and amplitudinal mapping of these responses was performed, demonstrating regions where quantitative delineation of cell stress was afforded.

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A Permeability Measurement of Small Unilamellar Vesicles by 6-Carboxyfluorescein$^*$

  • Lee, Choong-Hee;Choi, Myung-Un
    • Bulletin of the Korean Chemical Society
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    • v.5 no.4
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    • pp.154-158
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    • 1984
  • In order to characterize the permeability of small unilamellar vesicles (SUV), efflux of 6-carboxyfluorescein (6-CF) from the vesicles was monitored spectrophotofluorometrically. Since the entrapped highly quenched 6-CF (200 mM) became fluorescent upon release from the vesicles, the 6-CF could be used as an efflux probe. SUV containing entrapped 6-CF was prepared from egg phosphatidylcholine and separated by gel filtration on Sepharose 4B. Observed change of relative fluorescent intensity with time was sigmoidal. From this curve, the parameter of permeability was determined either by half-time or a released amount per unit time from the initial slope. Half-time of efflux of prepared SUV having 302 ng phospholipid/ml in 10 mM Tris-HCl buffer pH 7.4 was 21.0 min at $37{\circ}C$. Various factors which could affect the half-time were examined including temperature, pH, salt, and vesicle concentration. In particular the effect of vesicle concentration on the efflux revealed that the permeability can be a function of the concentration.

Detection of Chromosomal Rearrangements by Chromium in Human Lymphocyte Using Fluorescence in situ Hybridization (FISH) with Triple Combination of Composite whole Chromosome Specific Probe (FISH(fluorescence in situ hybridization)를 이용하여 분석한 크롬에 의해 유발된 염색체 이상)

  • 정해원;김수영;맹승희;이용묵;유일재
    • Environmental Mutagens and Carcinogens
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    • v.19 no.1
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    • pp.14-19
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    • 1999
  • Chromosome rearrangements induced in human lymphocyte after in vitro exposure to chromium were analysed by the use of fluorescence in situ hybridization(FISH) with triple combination of composite whole chromosome-specific probe for chromosome 1, 2 and 4. Chromosome aberrations was scored by the Protocol for Aberration Identification and Nomenclature Terminology (PAINT). Stable translocation was the most frequent type of aberrations and dicentrics and insertions were also observed. Chromium treatment enhanced the frequencies of stable translocations and color junctions in a dose-dependent manners, but no distinct increase of dicentrics and insertions was seen. The ratio of the yields of translocation to the yields of dicentric varied between 13 to 27. The presents results demonstrate fluorescent in situ hybridization (FISH) is useful for detecting chromosomal rearrangements induced by chromium.

Hybridization by an Electrical Force and Electrochemical Genome Detection Using an Indicator-free DNA on a Microelectrode-array DNA Chip

  • Choi, Yong-Sung;Lee, Kyung-Sup;Park, Dae-Hee
    • Bulletin of the Korean Chemical Society
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    • v.26 no.3
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    • pp.379-383
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    • 2005
  • This research aims to develop DNA chip array without an indicator. We fabricated microelectrode array by photolithography technology. Several DNA probes were immobilized on an electrode. Then, indicator-free target DNA was hybridized by an electrical force and measured electrochemically. Cyclic-voltammograms (CVs) showed a difference between DNA probe and mismatched DNA in an anodic peak. Immobilization of probe DNA and hybridization of target DNA could be confirmed by fluorescent. This indicator-free DNA chip microarray resulted in the sequence-specific detection of the target DNA quantitatively ranging from $10^{-18}\;M\;to\;10^{-5}$ M in the buffer solution. This indicator-free DNA chip resulted in a sequence-specific detection of the target DNA.

PHOTOPHYSICAL AND OPTICAL PROBE PROPERTIES OF 1-(p-N,N-DIMETHYLAMINOPHENYL)-4-PHENYL-2-METHYL-1E,3E-BUTADIENE

  • Singh, A.K.;Krishna, T.S.R.
    • Journal of Photoscience
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    • v.4 no.1
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    • pp.1-5
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    • 1997
  • A hitherto unknown diphenylbutadiene analog viz. 1-(p-N,N-dimethylaminophenyl)-4-phenyl-2-methyl-1E,3E-butadiene (10) has been prepared and its absorption, excitation, and fluorescent emission properties in different media including various organic solvents and aqueous bovine serum albumin (BSA) have been studied. For comparision, these properties have also been investigated for the parent diphenylbutadiene (2). Diene 10 exhibits solvent polarity/polarizability-sensitive fluorescence properties ($\lambda$$_{max}$, $\Phi$$_f$, $\tau$$_f$, K$_f$, f). It also binds to the hydrophobic domains of aqueous bovine serum albumin (BSA) with a binding constant of 3.89 x 10$^4$ M$^{-1}$. The relative fluorescence quantum yield of 10 increases, while, the fluorescence lifetime decreases with increasing concentration of-BSA. The results highlight the polar character of the singlet excited state of diphenylpolyenes and the utility of 10 as fluorescence probe for studying microenvironments of organized assemblies and biological supramolecular structures.

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