• Title/Summary/Keyword: Fluorescence Detection

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Single-molecule Detection of Fluorescence Resonance Energy Transfer Using Confocal Microscopy

  • Kim, Sung-Hyun;Choi, Don-Seong;Kim, Do-Seok
    • Journal of the Optical Society of Korea
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    • v.12 no.2
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    • pp.107-111
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    • 2008
  • We demonstrated single-molecule fluorescence resonance energy transfer (FRET) from single donor-acceptor dye pair attached to a DNA with a setup based on a confocal microscope. Singlestrand DNAs were immobilized on a glass surface with suitable inter-dye distance. Energy transfer efficiency between the donor and the acceptor dyes attached to the DNA was measured with different lengths of DNA. Photobleaching of single dye molecule was observed and used as a sign of single-molecule detection. We could achieve high enough signal-to-noise ratio to detect the fluorescence from a single-molecule, which allows real-time observation of the distance change between single dye pairs in nanometer scale.

Recent Progress in Multiplexed Detection of Biomarkers Based on Quantum Dots (양자점 기반 다중 바이오마커 검출법의 연구동향)

  • Kim, Yerin;Choi, Yu Rim;Kim, Bong-Geun;Na, Hyon Bin
    • Applied Chemistry for Engineering
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    • v.33 no.5
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    • pp.451-458
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    • 2022
  • Semiconductor quantum dots (QDs) are optical probes with excellent fluorescence properties. Therefore, they have been applied to various bio-medical imaging techniques and biosensors. Due to the unique optical characteristics of wide absorption and narrow fluorescence energy bands, multiple types of signals can be generated by the combination of fluorescence wavelengths from different QDs, which enables the simultaneous detection of more than two biomarkers. In this review, the advantages and applications of QDs and QD nanobeads (QBs) in multiple biomarker assays were described, and new developments or improvements in multiplexed biomarker detection techniques were summarized. In particular, recent reports were summarized, focusing on the design strategies in immunoassay construction and signal transducing materials for fluorescence-linked immunosorbent assays using QDs and immunochromatographic assays using QBs. New detection platforms will be developed for early diagnosis of diseases and other fields if multiplexed detection technologies of excellent accuracy and sensitivity are combined with artificial intelligence algorithms.

A STUDY ON THE EARLY DETECTION OF ENAMEL CARIES BY THE LUMINESCENCE EXCITED BY ARGON LASER (아르곤 레이저 광감각법의 법랑질 우식증 조기탐지 효과에 관한 연구)

  • Lee, Nan-Young;Lee, Chang-Seop;Lee, Sang-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.24 no.1
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    • pp.313-324
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    • 1997
  • The aim of the present study was to describe an safe and convenient method for the early detection of enamel caries using laser fluorescence. Fluorescence from natually carious lesion of human teeth illuminated by an argon laser(488nm) was observed and photographed using barrier filter. Intact enamel was found to fluorescence with a yellowish light. Whereas, incipient caries lesions in the enamel were dearly visible as dark areas in contrast to the fluorescence surroundings. For evaluation of accuracy of this method, lesion depth measured by the laser fluorescence in light microscope was compared with that polarizing microscope. The results from the present study can be summarized as follows : 1. Enamel caries of smooth surface was observed as pale white spot and undefined outline in ordinary light. Whereas, lesion was clearly visible as dark spot in laser fluorescence. 2. There was no difference between ordinary light view and laser fluorescence in occlusal surface and interproximal surface. 3. There was no significant difference between the lesion depth observed by laser fluorescence with light microscope and polarizing microscope. Apparent correlation exists between two groups.

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A Pyrenylboronic Acid-based Fluorescence Sensor for Highly Efficient Detection of Mercury(II) Ions (효율적인 수은이온 검출을 위한 피렌-보론산 기반의 형광센서 개발)

  • Lee, Seung Yeob;Lee, Seoung Ho
    • Journal of Environmental Science International
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    • v.29 no.2
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    • pp.201-207
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    • 2020
  • A new chemosensor based on a self-assembled system has been devised to detect Hg2+ions efficiently. We demonstrated that the amphiphilic building blocks consisting of pyrene and boronic acid (1) aggregate in aqueous solutions and provide an outstanding sensing platform for sensitive detection. The self-assembled 1 exhibited high selectivity and sensitivity for Hg2+ion detection via fluorescence quenching, where the Hg2+ion detection ensued from a fast transmetallation of 1. The Stern-Volmer (SV) quenching constant for its fluorescence quenching by Hg2+ions was approximately 1.58 × 108 M-1. In addition, self-assembled 1 exhibited excellent sensing abilities at nano-molar concentration levels when tap water and freshwater samples were contaminated with of Hg2+ ions.

Determination of Glyphosate in Whole Blood by HPLC-fluorescence Detection (HPLC 형광검출법에 의한 Glyphosate의 혈중농도 측정)

  • 이상기;김기욱;양자열;인상환;이수연
    • YAKHAK HOEJI
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    • v.45 no.4
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    • pp.347-351
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    • 2001
  • A rapid and sensitive method for the determination of glyphosate, a phosphated amino acid herbicide, in whole blood is presented. After removal of protein, the whale blood was purified by using the anion exchange resin (Dowex 1), and derivatized with 9-fluorenylmethyl chloroformate (FMCL). Derivatized glyphosate from blood sample was injected onto a Whatman partisil 10SAX column and separated with 0.1M phosphate buffer (pH 2.5) and acetonitrile (ratio=3:1). The high performance liquid chromatography-fluorescence detection gave the detection limit of 86pg and linearity of 0.9999 in the range of 0.25 $\mu$g/ml and 25 $\mu$g/ml. The recoveries of glyphosate added to the blood samples were ranged from 75.3% to 100.4% compared to the samples prepared in water. The derivatized glyphosate was stable at various acidity and temperature. This method has been successfully applied to the blood samples of lethal intoxication with the herbicide glyphosate.

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Photoreactivity of Anthraquinones for the Analysis of Ginsenosides Using Photoreduction Fluorescence Detection-HPLC

  • Park, Man-Ki;Kim, Bak-Kwang;Park, Jeong-Hill;Shin, Young-Geun;Cho, Kyung-Hee;Do, Young-Mi
    • Archives of Pharmacal Research
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    • v.19 no.6
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    • pp.562-565
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    • 1996
  • The photoreactivity of twelve anthraquinone derivatives was examined to evaluate its usefulness as a photo-reagent for the analysis of ginsenosides using photoreduction fluorescence (PRF) detection method. Among the tested compounds, 2-tert-butylandthraquinone (TBAQ), 2-chloroanthraquinone (CAQ) and anthraquinone (AQ) showed good characteristics as photoreagents. The detection limits of ginsenoside $Rg_{1}$PRF-HPLC method using TBAQ, CAQ or AQ as a photo-reagent were found to be ca. 35 ng, 50 ng and 50 ng, respectively.

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Analysis of Saikosaponins by HPLC with Photoreduction Fluorescence Detection (광반응 HPLC를 이용한 시호 사포닌의 분석)

  • Shin, Young-Geun;Cho, Kyung-Hee;Kwon, Soo-Jin;Do, Young-Mi;Hwang, Gwi-Seo;Park, Jeong-Hill;Park, Man-Ki
    • YAKHAK HOEJI
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    • v.40 no.1
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    • pp.41-45
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    • 1996
  • A high performance liquid chromatography using photoreduction fluorescence detection was described for the analysis of saikosaponins. Saikosaponins were separated on an $NH_2$ column using acetonitrile and aqueous 2-tert-butylanthraquinone(t-BAQ) as mobile phase. Column effluent was passed through a 40cm PTFE capillary tube coiled around a 10W UV lamp to reduce t-BAQ to a highly fluorescent dihydroxyanthracene derivative which was detected by a fluorescence detector. The optimal concentration of t-BAQ was found to be $6{\times}10^{-5}M$ and the optimal reaction time to be 2 seconds. The detection limit for saikosaponin a and d by this method was found to be about 280ng and 80ng. The dynamic linear range was over two orders and the correlation coefficient of the calibration curve of them was 0.998.

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Fabrication and characteristic evaluation of microfluidics chip integrated OLED for the light sources (OLED광원이 집적화된 마이크로 플루이딕칩의 제작 및 특성 평가)

  • Kim, Young-Hwan;Han, Jin-Woo;Kim, Jong-Yeon;Kim, Byoung-Yong;Seo, Dae-Shik
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2007.06a
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    • pp.377-377
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    • 2007
  • A simplified integration process including packaging is presented, which enables the realization of the portable fluorescence detection system. A fluorescence detection microchip system consisting of an integrated PIN photodiode, an organic light emitting diode (OLED) as the light source, an interference filter, and a microchannel was developed. The on-chip fluorescence detector fabricated by poly(dimethylsiloxane) (PDMS)-based packaging had thin-film structure. A silicon-based integrated PIN photo diode combined with an optical filter removed the background noise, which was produced by an excitation source, on the same substrate. The active area of the finger-type PIN photo diode was extended to obtain a higher detection sensitivity of fluorescence. The sensitivity and the limit of detection (LOD S/N = 3) of the system were $0.198\;nA/{\mu}M$ and $10\;{\mu}M$, respectively.

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Development and Characterization of Finger-type PIN Photodiode for Fluorescence Detection of RNA (RNA 형광 검출을 위한 Finger형 PIN 광다이오드의 제작 및 평가)

  • Kim, Ju-Hwan;Oh, Myung-Hwan;Ju, Byeong-Kwon
    • Journal of Sensor Science and Technology
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    • v.13 no.2
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    • pp.85-89
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    • 2004
  • This paper represents the development of high sensitivity photo-sensor for the fluorescence detection in the integrated biological analysis system. The finger-type PIN photodiodes were fabricated as the photo-sensor, and had a high sensitivity ($I_{light}/I_{dark}$ = 8720). The interference filter consisted of $TiO_{2}$ and $SiO_{2}$ was directly deposited on the photodiodes. Deposited filter with 95.5% reflection under 532 nm and 98% transmission over 580 nm exceedingly decreased the magnitude of background signal in the detection. The PDMS micro-fluidic channels are bonded on the photodiode by $O_{2}$ plasma treatment. The detection current was proportional to two primary parameters (light intensity, concentration), and the on-chip detection system could detect fluorescence signals down to 100 nM concentration (LOD = Limit of detection of rhodamine).

Single C-Reactive Protein Molecule Detection on a Gold-Nanopatterned Chip Based on Total Internal Reflection Fluorescence

  • Heo, Yunmi;Lee, Seungah;Lee, Sang-Won;Kang, Seong Ho
    • Bulletin of the Korean Chemical Society
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    • v.34 no.9
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    • pp.2725-2730
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    • 2013
  • Single C-reactive protein (CRP) molecules, which are non-specific acute phase markers and products of the innate immune system, were quantitatively detected on a gold-nanopatterned biochip using evanescent field-enhanced fluorescence imaging. The $4{\times}5$ gold-nanopatterned biochip (spot diameter of 500 nm) was fabricated by electron beam nanolithography. Unlabeled CRP molecules in human serum were identified with single-molecule sandwich immunoassay by detecting secondary fluorescence generated by total internal reflection fluorescence (TIRF) microscopy. With decreased standard CRP concentrations, relative fluorescence intensities reduced in the range of 33.3 zM-800 pM. To enhance fluorescence intensities in TIRF images, the distance between biochip surface and CRP molecules was optimally adjusted by considering the quenching effect of gold and the evanescent field intensity. As a result, TIRF only detected one single-CRP molecule on the biochip the first time.