• Title/Summary/Keyword: Fluorescence Measurement

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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 $P_{i}$.

Micro-LIF Measurement in a Micro-channel Using an Micro Laser Light Sheet (마이크로 레이저 평면빔을 이용한 마이크로채널 내에서의 Micro-LIF 측정)

  • Yoon, Sang-Youl;Kim, Jae-Min;Kim, Su-Hun;Kim, Kyung-Chun
    • Proceedings of the KSME Conference
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    • 2004.11a
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    • pp.1540-1545
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    • 2004
  • Measurement of concentration fields in a micro-channel is the crucial technology in the area of Lab-on-a-chip to be used for various bio-chemical applications. It is wel-known that the only possible way to measure the concentration field in the micro-channel is using micro-LIF(Laser Induced Fluorescence) method. However, an accurate concentration field at a given cross plane in a micro-channel has not been made so far due to the limit of light illumination. The present study demonstrates a novel method to provide an ultra thin laser sheet beam having 5 microns thickness by a micro focus laser line generator. Nile Blue A was used as fluorescent dye for LIF measurement. The laser sheet beam illuminates an exact plane of concentration measurement in the micro-channel to increase the signal to noise ratio and reduce the depth uncertainty considerably.

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Study on the Detachment of Fluorescent Whitening Agents from White Waster Papers (인쇄용지 재활용을 위한 형광증백제 탈착에 대한 기초연구)

  • Lee, Ji Young;Kim, Chul Hwan;Kim, Eun Hea;Park, Tae Ung
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.48 no.2
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    • pp.5-12
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    • 2016
  • White waste papers are very important resources in the paper industry, but their use is limited because of the residual of fluorescent whitening agent (FWA). So the removal of FWAs from waste paper is an important task in the recycling process to improve the use of recycled resources. In this study, we focused on the FWAs used for surface treatments and carried out physical and chemical treatments to remove them from white waste papers. The white waste papers were disintegrated with a surfactant in different pH and temperature conditions, and then handsheets were made for the measurement of the fluorescence index, which is proportional to the amount of FWAs on papers. The effect of the flotation process on the removal of FWAs after disintegration was also investigated. The fluorescence index decreased as the disintegration time increased, but over a relatively long time, the fluorescence index increased again, which indicated the readsorption of the FWAs detached from the cellulosic fibers of the white waste papers. The lowest fluorescence index was shown when the waste papers were disintegrated with a 0.3% surfactant addition at pH 10 and at $45^{\circ}C$. However, the flotation treatment was not effective, because the flotation induced contact between the detached FWAs and the cellulosic fibers, and re-adsorption occurred.

Assessment of PLLIF Measurement for Spray Mass Distribution of Like-Doublet Injector (Like-Doublet 인젝터의 분무 질량분포 측정을 위한 PLLIF기법의 신뢰성 평가)

  • Jung Kihoon;Koh Hyeonseok;Yoon Youngbin
    • Journal of the Korean Society of Visualization
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    • v.1 no.1
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    • pp.98-106
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    • 2003
  • A PLLIF (Planar Liquid Laser Induced Fluorescence) technique has been known to be a useful tool for the measurement of the spray patterns for various spray injectors because it can obtain two-dimensional images with high spatial resolutions without any intrusion on the spray field. In case of dense spray, however, the secondary emission as well as the extinction of an incident laser beam or a fluorescence signal can cause errors in quantifying a mass distribution. Unfortunately, a like-doublet injector which has a dense spray zone at the center may not be a good example or the application of the PLLIF technique. Therefore, we took PLLIF data for the like-doublet injector with a 12 bit color CCD camera by varying laser power, and then assessed their accuracy by comparing with the data obtained with a mechanical patternator and a PDPA (Phase Doppler Particle Analyzer). The experimental results showed that the gray level of fluorescence signal increases nonlinearly due to a secondary emission at the dense spray zone but this nonlinearity can be avoided by reducing the incident laser beam power. In addition, the mass flux distribution of the spray could be obtained by using the mass concentration data from PLLIF technique and the velocity profiles of liquid drops, and this distribution showed good agreement with that of mechanical pattemator. Therefore, it is possible that the PLLIF technique can be successfully applied to finding the mass distributions of like-doublet injectors.

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Relaxation Process of the Photoexcited State and Singlet Oxygen Generating Activity of Water-soluble meso-Phenanthrylporphyrin in a DNA Microenvironment

  • Hirakawa, Kazutaka;Ito, Yusuke;Yamada, Takashi;Okazaki, Shigetoshi
    • Rapid Communication in Photoscience
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    • v.3 no.4
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    • pp.81-84
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    • 2014
  • To examine the microenvironmental effect of DNA on the photosensitized reaction, the electron-donor-connecting porphyrin, meso-(9-phenanthryl)-tris(N-methyl-p-pyridinio) porphyrin (Phen-TMPyP), was synthesized. Phen-TMPyP can bind to oligonucleotides with two binding modes, depending on the DNA concentration. The fluorescence lifetime measurement of Phen-TMPyP shows a shorter component than that of the reference porphyrin without the phenanthryl moiety. However, the observed value is much longer than those of previously reported similar types of electron-donor-connecting porphyrins, suggesting that electron-transfer quenching by the phenanthryl moiety is not sufficient. The fluorescence quantum yield of Phen-TMPyP ($5{\mu}M$) decreased with an increase in DNA concentration of up to $5{\mu}M$ base pair (bp), possibly due to self-quenching through an aggregation along the DNA strand, increased with an increase in DNA concentration of more than $5{\mu}M$ bp and reached a plateau. The fluorescence quantum yield of Phen-TMPyP with a sufficient concentration of DNA was larger than that of the reference porphyrin. The singlet oxygen ($^1O_2$) generating activity of Phen-TMPyP was confirmed by the near-infrared emission spectrum measurement. The quantum yield of $^1O_2$ generation was decreased by a relatively small concentration of DNA, possibly due to the aggregation of Phen-TMPyP, and recovered with a sufficient concentration of DNA. The recovered quantum yield was rather smaller than that without DNA, indicating the quenching of $^1O_2$ by DNA. These results show that a DNA strand can stabilize the photoexcited state of a photosensitizer and, in a certain case, suppresses the $^1O_2$ generation.