• Title/Summary/Keyword: fluorescent detection

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A Label-Free Fluorescent Amplification Strategy for High-Sensitive Detection of Pseudomonas aeruginosa based on Protective-EXPAR (p-EXPAR) and Catalytic Hairpin Assembly

  • Lu Huang;Ye Zhang;Jie Liu;Dalin Zhang;Li Li
    • Journal of Microbiology and Biotechnology
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    • v.34 no.7
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    • pp.1544-1549
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    • 2024
  • This study presents a fluorescent mechanism for two-step amplification by combining two widely used techniques, exponential amplification reaction (EXPAR) and catalytic hairpin assembly (CHA). Pseudomonas aeruginosa (P. aeruginosa) engaged in competition with the complementary DNA in order to attach to the aptamer that had been fixed on the magnetic beads. The unbound complementary strand in the liquid above was utilized as a trigger sequence to initiate the protective-EXPAR (p-EXPAR) process, resulting in the generation of a substantial quantity of short single-stranded DNA (ssDNA). The amplified ssDNA can initiate the second CHA amplification process, resulting in the generation of many double-stranded DNA (dsDNA) products. The CHA reaction was initiated by the target/trigger DNA, resulting in the release of G-quadruplex sequences. These sequences have the ability to bond with the fluorescent amyloid dye thioflavin T (ThT), generating fluorescence signals. The method employed in this study demonstrated a detection limit of 16 CFU/ml and exhibited a strong linear correlation within the concentration range of 50 CFU/ml to 105 CFU/ml. This method of signal amplification has been effectively utilized to create a fluorescent sensing platform without the need for labels, enabling the detection of P. aeruginosa with high sensitivity.

Application of fluorescent particles as a tracer to detect the membrane surface damage in a pilot scale membrane bioreactor (형광입자를 이용한 분리막 표면 검측 방법의 파일럿 규모 플랜트 적용)

  • Kim, Choah;Kim, Hee Jun;Cho, Jinwoo
    • Journal of Korean Society of Water and Wastewater
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    • v.30 no.1
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    • pp.33-40
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    • 2016
  • In this study, a fluorescent silica nano particle is used as the surrogate for challenging test of membrane surface integrity. The particles are functionalized by a fluorescent dying agent so that as an ultraviolet light is imposed a bright fluorescent image from the particles can be taken. If a membrane surface is damaged and has a compromised part larger than the size of surrogate the fluorescent particles would pass through and contained in the permeate. An operator can directly notice whether the membrane surface is damaged or not by detecting a fluorescent image taken from the permeate. Additionally, the size of compromised part is estimated through analysing the fluorescent image in which we surmise the mass of particles included in the permeate by calculating an average RGB value of the image. The pilot scale experiments showed that this method could be applied successfully to determine if a membrane surface had a damaged parts regardless of the test condition. In the testing on the actual damaged area of $4.712mm^2$, the lowest error of estimating the damaged area was -1.32% with the surrogate concentration of 80 mg/L, flux of $40L/m^2/hr$ for 25 minutes of detection. A further study is still going on to increase the lowest detection limit and thus decrease the error of estimation.

A Coumarin-based Fluorescent Sensor for Selective Detection of Copper (II)

  • Wang, Jian-Hong;Guo, Xin-Ling;Hou, Xu-Feng;Zhao, Hui-Jun;Luo, Zhao-Yang;Zhao, Jin
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2400-2402
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    • 2014
  • Cu (II) detection is of great importance owing to its significant function in various biological processes. In this report, we developed a novel coumarin-based chemosensor bearing the salicylaldimine unit (2) for $Cu^{2+}$ selective detection. The results from fluorescence spectra demonstrated that the sensor could selectively recognize $Cu^{2+}$ over other metal cations and the detection limit is as low as $0.2{\mu}M$. Moreover, the confocal fluorescence imaging in HepG2 cells illustrated its potential for biological applications.

A New End of Lamp Life Detection Method for Fluorescent Lamps (새로운 형광램프 수명말기 현상 검출 방법)

  • Cho, Gye-Hyun
    • Journal of the Korean Institute of Illuminating and Electrical Installation Engineers
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    • v.21 no.7
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    • pp.1-5
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    • 2007
  • This paper presents the new detection method for the end of life on fluorescent lamps. At the end of lamp life, the lamp voltage and current asymmetrically increase and decrease more than normal state. If the ballast system does not have the protection function especially for T4 and T5 lamps, we may see the melting socket which is connected to the end of the lamp. To protect from this kind of abnormal status is the most important thing in the ballast system that has very old lamps.

Development of a Fluorescent Sensor Based on Resazurin and Hydrotalcite for the Determination of Ethanol in Alcoholic Beverages

  • Hong Dinh Duong;Juyeon Kim;Jong Il Rhee
    • Journal of Sensor Science and Technology
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    • v.33 no.2
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    • pp.70-77
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    • 2024
  • In this study, a fluorescent ethanol sensor is developed to determine the ethanol concentration in the liquid phase. The sensor is developed using a complex of resazurin (RA)/resorufin (RO) and a hydrotalcite (HT) catalyst in a sol-gel matrix of methyltrimethoxysilane (MTMS) to produce a fluorescent ethanol-sensing membrane (RA/RO*HT membrane). The operation mechanism of the RA/RO*HT membrane is based on (i) the oxidation of ethanol to acetaldehyde and (ii) the reduction of RA to RO, through electron flows followed by EtOH ↔ HT ↔ RA/RO ↔ EtOH interactions. These possible redox reactions can lead to an increased fluorescence intensity of the RA/RO*HT membrane as the ethanol concentration increases. The RA/RO*HT membrane shows a linear detection range of 1-20 vol.% EtOH with limit of detection (LOD) of 0.178%. Additionally, the RA/RO*HT membrane has high sensitivity and accuracy for determining the alcohol content in several Korean alcoholic beverages.

Eastern Staining: A Simple Recombinant Protein Detection Technology Using a Small Peptide Tag and Its Counter Partner Which is a Fluorescent Compound

  • Lee, Jae-Jung;Kim, Jun-Young;Zhai, Duanting;Yun, Seong-Wook;Chang, Young-Tae
    • Interdisciplinary Bio Central
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    • v.4 no.2
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    • pp.5.1-5.9
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    • 2012
  • Small peptide tags such as c-myc, HA, or FLAG tag have facilitated efficient Western-blotting of proteins of interest especially when specific antibodies for the proteins are not available. However, the conventional Western-blotting requires the multi-steps process taking at least several hours up to two days. With examples of various applications, here we show a convenient and time-saving method for protein detection which employs a fluorescent chemical BDED and its binding peptide RC-tag. And we propose "Estern staining", as a standard term for protein detection method using fluorescent chemicals and their binding small peptide tags. Eastern staining may substitutes for the time-consuming "immuno-staining" in many versatile applications.

Specific Detection of DNA Using Quantum Dots and Magnetic Beads for Large Volume Samples

  • Kim, Yeon-Seok;Kim, Byoung-Chan;Lee, Jin-Hyung;Kim, Jung-Bae;Gu, Man-Bock
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.5
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    • pp.449-454
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    • 2006
  • Here we present a sensitive DNA detection protocol using quantum dots (QDs) and magnetic beads (MBs) for large volume samples. In this study, QDs, conjugated with streptavidin, were used to produce fluorescent signals while magnetic beads (MBs) were used to isolate and concentrate the signals. The presence of target DNAs leads to the sandwich hybridization between the functionalized QDs, the target DNAs and the MBs. In fact, the QDs-MBs complex, which is bound using the target DNA, can be isolated and then concentrated. The binding of the QDs to the surface of the MBs was confirmed by confocal microscopy and Cd elemental analysis. It was found that the fluorescent intensity was proportional to concentration of the target DNA, while the presence of non-complementary DNA produced no significant fluorescent signal. In addition, the presence of low copies of target DNAs such as 0.5 pM in large volume samples up to 40mL was successfully detected by using a magnet-assisted concentration protocol which consequently results in the enhancement of the sensitivity more than 100-fold.