• Title/Summary/Keyword: Real time fluorescence measurement

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Development of a pH/dissolved- oxygen Monitoring System Using HPTS and Rudpp (HPTS, Rudpp를 활용한 pH 및 용존산소 모니터링 시스템 연구)

  • Dong Hyuk Jeong;Daewoong Jung
    • Journal of Sensor Science and Technology
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    • v.32 no.2
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    • pp.82-87
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    • 2023
  • This study proposes a pH-dissolved-oxygen monitoring system using 8-HydroxyPyrene-1,3,6-trisulfonic acid Trisodium Salt (HPTS) and tris(4,7-diphenyl-1,10-phenanthroline)Ruthenium(II) chloride (Rudpp). Commercial water-quality sensors are electrochemical devices that require frequent calibration and cleaning, are subject to high maintenance costs, and have difficulties conducting measurements in real-time. The proposed pH-dissolved-oxygen monitoring system selects a thin-film sensing layer to measure the change in fluorescence intensity. This change in fluorescence intensity is based on reactions with hydrogen ions in an aqueous solution at a given pH and specific amount of dissolved oxygen. The change in fluorescence intensity is then measured using light-emitting diodes and photodiodes in response to HPTS and Rudpp. This method enables the development of a relatively small, inexpensive, and real-time measureable water-quality measurement system.

Real Time Scale Measurement of Inorganic Phosphate Release by Fluorophore Labeled Phosphate Binding Protein (형광단이 붙어 있는 인산결합 단백질에 의한 인산 배출의 실시간 측정)

  • Jeong Yong-Joo
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.935-940
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    • 2005
  • Fluorescence change of coumarin labeled phosphate binding protein (PBP-MDCC) was monitored to measure the amount of released inorganic phosphate ($P_{i}$) during nucleoside triphosphate (NTP) hydrolysis reaction. After purification of PBP-MDCC, fluorescence emission spectra showed that fluorescence responded linearly to $P_{i}$ up to about 0.7 molar ratio of $P_{i}$ to protein. The correlation of fluorescence signal and $P_{i}$ standard was measured to obtain [$P_{i}$] - fluorescence intensity standard curve on the stopped-flow instrument. When T7 bacteriophage helicase, double-stranded DNA unwinding enzyme using dTTP hydrolysis as an energy source, reacted with dTTP, the change of fluorescence was able to be converted to the amount of released $P_{i}$ by the $P_{i}$ standard curve. $P_{i}$ release results showed that single-stranded Ml3 DNA stimulated dTTP hydrolysis reaction several folds by T7 helicase. Instead of end point assay in NTP hydrolysis reaction, real time $P_{i}$-release assay by PBP-MDCC was proven to be very easy and convenient to measure released $P_{i}$.

Development of Image-based Fluorescence Photobleaching Technique for Measuring Macromolecule Diffusion in Biological Porous Medium (생체 다공성 매질에서 분자 확산 측정을 위한 영상 기반 형광 광표백 기법 개발)

  • Lee, Dong-Hee;Lee, Jeong-Hoon;Park, Choon-Ho;Kim, Jung-Kyung
    • Journal of the Korean Society of Visualization
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    • v.7 no.1
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    • pp.9-13
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    • 2009
  • Fluorescence recovery after photobleaching (FRAP) has been widely used for the measurement of molecular diffusion in living cells and tissues. We developed an image-based FRAP (iFRAP) technique using a modified real-time microscope and a 488 nm Ar-ion laser. A fractional intensity curve was obtained from the time-lapse images of fluorescence recovery in the bleached spot to determine the diffusion coefficient of fluorescently labeled macromolecules in porous medium. We validated iFRAP through experiments with agar gels (0.5% and 1.5% w/v) containing FITC-Dextrans (10, 70 and 500 kDa MW). Further validation was performed by a Monte Carlo approach, where we simulated the three-dimensional random walk of macromolecules in agar gel model. Diffusion coefficients were deduced from the mean square displacement curves and showed good agreements with those measured by iFRAP.

The Study of Optical Biopsy‘s Usefulness in Radiotherapy (방사선 치료에서 광학적 생검의 유용성에 관한 연구)

  • ;;Muller M.G.,;Feld M.S.
    • Progress in Medical Physics
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    • v.12 no.1
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    • pp.9-17
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    • 2001
  • The prior purpose of this study is to introduce a optical biopsy and evaluate whether the optical biopsy, real-time, non-invasive technique, is a reliable tool to assess response to radiotherapy Four healthy volunteers, and four patients with inflammatory conditions of the oral cavity participated on the study. was obtained from each person enrolled in the study. Using FastEEM(Ercited Emission Matrix) as a optical biopsy tool, normal and tumor spectra are taken from the normal and the tumor regions. And then second optical biopsy are taken from the tumor regions in 4 patients with time delay at 7days.. Using a diagnostic algorithm, made by Gillenwater based on spectra excited at 337nm The Optical Biopsy turned out to be more suited for tumor diagnostic resulting in significant difference fluorescence spectra. The fluorescence intensity of cancerous tissue showed a higher position. The second fluorescence intensity of optical biopsy of cancerous oral tissue has more smaller than the first result. I conclude that optical biopsy, which technique don't need to remove tissue sample from body, and is a real time , and non-invasive measurement is a reliable tool to access to radiotherapy because FastEEM can do measure the variation of the tissue composition chemical, biological, and morphological after radiotherapy. Based on the fluorescence spectrum are taken from the optical biopsy in normal and tumor spectra as well as tumor spectra after 7days.

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A Study on the Correlation between the Harmful Cyanobacterial Density and Phycocyanin Concentration at Recreational Sites in Nakdong River (낙동강 친수활동구간 유해 남조류 분포와 피코시아닌(Phycocyanin) 농도 상관성에 관한 연구)

  • Hyo-Jin Kim;Min-Kyeong Kim
    • Journal of Korean Society on Water Environment
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    • v.39 no.6
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    • pp.451-464
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    • 2023
  • Harmful cyanobacterial monitoring is time-consuming and requires skilled professionals. Recently, Phycocyanin, the accessory pigment unique to freshwater cyanobacteria, has been proposed as an indicator for the presence of cyanobacteria, with the advantage of rapid and simple measurement. The purpose of this research was to evaluate the correlation between the harmful cyanobacterial cell density and the concentration of phycocyanin and to consider how to use the real-time water quality monitoring system for algae bloom monitoring. In the downstream of the Nakdong River, Microcystis spp. showed maximum cell density (99 %) in harmful cyanobacteria (four target genera). A strong correlation between phycocyanin(measured in the laboratory) concentrations and harmful cyanobacterial cell density was observed (r = 0.90, p < 0.001), while a weaker relationship (r = 0.65, p < 0.001) resulted between chlorophyll a concentration and harmful cyanobacterial cell density. As a result of comparing the phycocyanin concentration (measured in submersible fluorescence sensor) and harmful cyanobacterial cell density, the error range increased as the number of cyanobacteria cells increased. Before opening the estuary bank, the diurnal variations of phycocyanin concentrations did not mix by depth, and in the case of the surface layer, a pattern of increase and decrease over time was shown. This study is the result of analysis when Microcystis spp. is dominant in downstream of Nakdong River in summer, therefore the correlation between the harmful cyanobacteria density and phycocyanin concentrations should be more generalized through spatio-temporal expansion.

Engineering of Recombinant Escherichia coli Towards Methanol Sensing Using Methylobacterium extroquens Two-component Systems

  • Selvamani, Vidhya;Ganesh, Irisappan;Chae, Sowon;Maruthamuthu, Murali kannan;Hong, Soon Ho
    • Microbiology and Biotechnology Letters
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    • v.48 no.1
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    • pp.24-31
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    • 2020
  • Five genes (mxbDM, mxcQE and mxaB) are responsible for the transcription of methanol oxidation genes in Methylobacterium strains. Among these, MxbDM and MxcQE constitute the two-component system (TCS) regulating methanol metabolism. In this study, we integrated the methanol-sensing domain of MxbD and MxcQ with the EnvZ/OmpR from Escherichia coli. The domain-swapping strategy resulted in chimeric histidine kinases (HK's) MxbDZ and MxcQZ AM1 containing recombinant E. coli. Real-time quantitative PCR was used to monitor OmpC expression mediated by the chimeric HK and response regulator (RR) OmpR. Further, an ompC promoter based fluorescent biosensor for sensing methanol was developed. GFP fluorescence was studied both qualitatively and quantitatively in response to environmental methanol. GFP measurement also confirmed ompC expression. Maximum fluorescence was observed at 0.05% methanol and 0.01% methanol using MxbDZ and MxcQZ AM1, respectively. Thus the chimeric HK containing E. coli were found to be highly sensitive to methanol, resulting in a rapid response making them an ideal sensor.

Study on Improvement of Signal to Background Ratio of Laser-based Fluorescence Imaging System (레이저 기반 형광 영상 시스템의 Signal to Background Ratio 향상 연구)

  • Kim, J.H.;Jeong, M.Y.
    • Journal of the Microelectronics and Packaging Society
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    • v.27 no.4
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    • pp.107-111
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    • 2020
  • Recently, as an aging society progresses, a lot of interest in health and diagnosis is increasing, As the field of various bio-imaging systems for guided surgery capable of accurate diagnosis has emerged as important, a Fluorescence imaging system capable of accurate measurement and real-time confirmation has emerged as an important field. Fluorescence images currently being used are mainly in the NIR-I band, but many studies are in progress in the NIR-II band in order to improve resolution and confirm fluorescence deeply and accurately. In this paper, the difference between NIR-I and NIR-II, optical characteristics, and SBR (signal to background ration) of a fluorescent imaging system, was investigated using the finite element (FEM) method. After confirming, it was confirmed that the SBR was 16.2 times higher in the NIR-II area than in the NIR-I by making the skin phantom and measuring the fluorescence. It is confirmed that the enhancement in SBR of the Fluorescence imaging system is more effective in the NIR-II region than in the NIR-I region and expected to be used in application fields such as guided surgery, bio-sensor and also device which can detect the defect of optical devices.

Analyses of Synchronous Fluorescence Spectra of Dissolved Organic Matter for Tracing Upstream Pollution Sources in Rivers (상류 오염원 추적을 위한 용존 유기물질 Synchronous 형광스펙트럼 분석 연구)

  • Hur, Jin;Kim, Mi-Kyoung;Park, Sung-Won
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.3
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    • pp.317-324
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    • 2007
  • Fluorescence measurements of dissolved organic matter(DOM) have the superior advantages over other analysis tools for applying to water quality management. A preliminary study was conducted to test the feasibility of applying synchronous fluorescence measurements for tracing and monitoring pollution sources in a small stream located in an upstream area of the Sooyoung watershed in Busan. The water quality of the small stream is affected by leachate from sawdust pile and discharge of untreated sewage. The sampling sites included an upstream site, two pipes discharging untreated sewage, leachate from sawdust, and a downstream site. Of the five field samples, the leachate was distinguished from the other samples by a high peak at a lower wavelength range and a blunt peak at 350nm, suggesting that synchronous fluorescence can be used as a discrimination tool for monitoring the pollution. The efficacy of various indices derived from the spectral features to discriminate the pollution source was tested for well-defined mixture of the sawdust leachate and the upstream stream by comparing (1)the difference between measured values and those predicted based on mass balance and the characteristics of the two samples and (2)the linear correlations between index values and mass ratios of the sample mixtures. Of various discrimination indices selected, fluorescence intensities at 276 nm$({\Delta}\lambda=30nm)$and 347 nm$({\Delta}\lambda=60nm)$ were suggested as promising potential discrimination indices for the sawdust pollution source. Despite the limited number of samples and the study area, this study illustrates the evaluation process that should be followed to develop rapid, low-cost discrimination indices to monitor pollution sources based on end member mixing analyses.

Distributions and Behaviors of H2O2 Above the Yellow Sea in the Years Between 2002 and 2004 (2002년에서 2004년 동안 서해상공에서 관측된 과산화수소의 농도분포 및 거동)

  • Kim Y.M.;Shin S.A.;Han J.S.;Lee M.H.;Kim J.A.
    • Journal of Korean Society for Atmospheric Environment
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    • v.21 no.6
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    • pp.689-697
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    • 2005
  • Hydrogen peroxide is a reservoir of OH radical which is the powerful oxidant in the atmosphere. Therefore, the status of the oxidizing atmosphere could be reflected on the concentration of $H_{2}O_{2}$. In this study, the distribution of $H_{2}O_{2}$ was determined during the intensive aircraft measurements over the Yellow sea in March, December 2002, April, November 2003 and March, October 2004. Flights covered from $124^{circ}E\;to\;129^{circ}E\;and\;35^{circ}N\;to\;37^{circ}N$, and extending to 3,000 m. The flight patterns were set properly to assess the altitudinal and longitudinal distribution for $H_{2}O_{2}$. $H_{2}O_{2}$ was extracted onto aqueous solution using a continuously flowing glass coil and analyzed by a high performance liquid chromatography (HPLC) accompanied with a fluorescence detector using postcolumn enzyme derivatization. Mixing ratios of $O_{3},\;NO_{x}\;and\;SO_{2}$ were measured in real time by commercial analysis instruments. Along the heights, the maximum concentration of $H_{2}O_{2}$ appeared around 1,500 m then gradually decreased with increasing altitude. The vertical behavior of ozone showed the similar trend to $H_{2}O_{2}$. The mean mixing ratio of $NO_{x}$ was about 2 ppbv and not showed clear vertical distribution patterns. The mean value of was the same as $NO_{x}$ however $SO_{2}$ appeared extreme concentration in low altitude. $H_{2}O_{2}\;and\;O_{3}$ showed even longitudinal distribution however $NO_{x}$ mixing ratio in land ($127^{circ}E$) was much higher than over the sea. $SO_{2}$ rather decreased with increasing longitude. $H_{2}O_{2}$ was in inverse proportion to $NO_{x}$ in spring and summer and $SO_{2}$ in spring, which indicated its significant role to NO and $SO_{2}$ oxidation pathways.