• Title/Summary/Keyword: fluorescent detection

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A comparative study of three different viability tests for chemically or thermally inactivated Escherichia coli

  • Park, Seon Yeong;Kim, Chang Gyun
    • Environmental Engineering Research
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    • v.23 no.3
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    • pp.282-287
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    • 2018
  • Three different methods of bacterial viability monitoring were compared to detect chemically or thermally inactivated Escherichia coli. Direct colony enumeration, live/dead bacterial cell staining with a fluorescent dye, and the dehydrogenase activity assay were compared with respect to their ease of use and time required to perform the three different tests. The green (live cell)/red (dead cell) ratio obtained from the fluorescent bacterial cell staining approach showed a linear relationship with the colony forming units; the result obtained with dehydrogenase was similar to those. The sensitivity of the monitoring methods to detect bacterial deactivation varied with different disinfection conditions. After thermal treatment, the sensitivity of the staining approach was lower, while that of the dehydrogenase activity assay was the highest. After chemical treatment, the sensitivity of detection for both methods was similar.

Screening of Green Fluorescent Protein Gene and Sexing by PCR in Bovine Embryos (소 수정란에서 Green Fluorescent Protein 유전자 검색 및 PCR에 의한 성감별)

  • Lee, H. J.;Kang, T. Y.;Rho, G. J.;Chae, Y. J.;Lee, H.;Choe, S. Y.
    • Journal of Embryo Transfer
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    • v.15 no.2
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    • pp.157-165
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    • 2000
  • The efficiency of transgenic livestock production could be improved by early screening of transgene-integration and sexing of embryos at preimplantational stages before trasferring them into recipients. We examined the effciency of multiplex PCR analysis for the simultaneous confirmation of the trasgene and sex during the preimplantational development of bovine embryos and the possibility of green fluorescent protein(GFP) gene as a non-invasive marker for the early screening of transgenic embryos. The GFP gene was microinjected into the male pronuclei of bovine zygotes produced in vitro. The injected zygotes were co-cultured in TCM-199 containing 10% FCS with boving oviductal epithelial cells in a 5% CO2 incubator. Seventeen(13.0%) out of 136 gene-injected bovine zygotes developed by multiplex PCR analysis and the expression of GFP was detected by observing green fluorescence in embryos under a fluorescent microscope. Eight(67%) of 12 embryos at 2-cell to blastocyst stage were positive in the PCR analysis, but only two(11.8%) of 17 blastocysts expressed the GFP gene. Their sex was determined as 7 female and 5 male embryos by the PCR analysis. The results indicate that the screening of GFP gene and sex in bovine embryos by PCR analysis and fluorescence detection could be a promisible method for the preselection of transgenic embryos.

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The Effectiveness of Environmental Management through Environmental Surveillance (환경감시를 통한 환경관리의 효과)

  • Mi Hyang Lee;Jae Yeun Kim;Sang Ha Kim
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.6
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    • pp.557-561
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    • 2023
  • This study aims to assess how effective environmental management can be accomplished through monitoring of environmental conditions in patient discharge rooms within healthcare facilities through direct observation and the use of fluorescent markers. From March to July 2013, this study evaluated 448 check-out beds in wards and intensive care units before and 494 after intensive environmental monitoring activities. The collected data were analyzed using the SPSS 21.0 program. According to the study's findings, direct observation increased from 95.2% prior to the implementation of intensive environmental monitoring activities to 98.9% following the implementation, which was statistically significant. The non-detection rate of fluorescent markers exhibited an increase from 96.1% prior to the commencement of intensive environmental monitoring activities to 98.0% following their implementation. However, it should be noted that this observed increase was not deemed statistically significant. In light of the results of this research, it is imperative to evaluate the effectiveness of environmental management by employing a variety of assessment methods, including direct observation and fluorescent markers.

Development of FISH Methods for Detection of Genus Bacillus (Bacillus 속 세균을 검출하기 위한 Fluorescent In Situ Hybridization 방법의 개발)

  • 홍선희;김옥선;송홍규;이동훈;안태석
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.204-208
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    • 2001
  • A technique for detection of Bacillus in soil and waste water treatment system was developed. Mixed col- tured solutions of 22 Bacillus strains were applied for selection of probe and pretreatment method for FISHAmong the three probes known as useful tool for FISH method,S-G-Bacill-0597-a-A-22 was best for detec-tion of Bacillus. Ninety five percent of DAPl count was observed with FISH method with S-G-Bacill-0597-a-A-22 probe. For increasing the permeability to Bacillus cell walls, pretreatment with lysozyme was betterthan that with lysozyme and SDS/DTT (dithiothreitol). Bacillus spore was not detected with FISH. So, Bacil-lus detection in ecosystem requires FISH with pretreatment of lysozyme and spore staining.

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Performance Evaluation of the Automated Fluorescent Immunoassay System Rotavirus Assay in Clinical Samples

  • Kim, Jae-Seok;Lee, Su-Kyung;Ko, Dae-Hyun;Hyun, Jungwon;Kim, Hyun Soo
    • Annals of Laboratory Medicine
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    • v.39 no.1
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    • pp.50-57
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    • 2019
  • Background: The Automated Fluorescent Immunoassay System (AFIAS) rotavirus assay (Boditech Med Inc., Chuncheon, Korea) is a new rapid antigen test for rotavirus detection. We evaluated the performance of this assay for detecting rotaviruses and their specific genotypes in clinical stool samples. Methods: AFIAS rotavirus assay was performed in 103 rotavirus-positive and 103 rotavirus-negative stool samples (confirmed by both PCR and ELISA), and its results were compared with those of PCR, ELISA, and immunochromatographic assay (ICA). We evaluated diagnostic sensitivity/specificity, the detectability of rotavirus subtypes, lower limit of detection (LLOD), reproducibility, cross-reactivity, and interference of AFIAS rotavirus assay. Results: Based on PCR and ELISA results, diagnostic sensitivity and specificity of the AFIAS rotavirus assay were both 99.0%. LLOD results showed that the AFIAS assay had sensitivity similar to or greater than ICA and ELISA. High reproducibility was confirmed, and no cross-reactivity or interference was detected. This assay could detect genotypes G1P[8], G2P[4], G3P[8], G4P[6], G4P[8], G8P[4], G8P[8], G9P[4], and G9P[8]. Conclusions: The AFIAS rotavirus assay showed high reproducibility, sensitivity, and specificity as well as excellent agreement with ELISA, PCR, and ICA. It detected the most common as well as unusual genotypes of rotavirus prevalent in Korea. It could be a useful onsite assay for rapid, convenient, and cost-effective detection of rotavirus infection.

Enhancement of Photoluminescence by Ag Localized Surface Plasmon Resonance for Ultraviolet Detection

  • Lyu, Yanlei;Ruan, Jun;Zhao, Mingwei;Hong, Ruijin;Lin, Hui;Zhang, Dawei;Tao, Chunxian
    • Current Optics and Photonics
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    • v.5 no.1
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    • pp.1-7
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    • 2021
  • For higher sensitivity in ultraviolet (UV) and even vacuum ultraviolet (VUV) detection of silicon-based sensors, a sandwich-structured film sensor based on Ag Localized Surface Plasmon Resonance (LSPR) was designed and fabricated. This film sensor was composed of a Ag nanoparticles (NPs) layer, SiO2 buffer and fluorescence layer by physical vapour deposition and thermal annealing. By tuning the annealing temperature and adding the SiO2 layer, the resonance absorption wavelength of Ag NPs matched with the emission wavelength of the fluorescence layer. Due to the strong plasmon resonance coupling and electromagnetic field formed on the surface of Ag NPs, the radiative recombination rate of the luminescent materials and the number of fluorescent molecules in the excited state increased. Therefore, the fluorescent emission intensity of the sandwich-structured film sensor was 1.10-1.58 times at 120-200 nm and 2.17-2.93 times at 240-360 nm that of the single-layer film sensor. A feasible method is provided for improving the detection performance of UV and VUV detectors.

Evaluation of Selective Media Containing Iron Source and Alpha-Glucosidase Substrates for Enterobacter sakazakii (Cronobacter spp.) Detection

  • Chon, Jung-Whan;Seo, Kun-Ho;Yim, Jin-Hyeok;Bae, Dongryeoul;Kim, Binn;Kim, Tae-Jin;Jeong, Dongkwan;Song, Kwang-Young
    • Journal of Dairy Science and Biotechnology
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    • v.39 no.1
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    • pp.9-19
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    • 2021
  • Enterobacter sakazakii (Cronobacter spp.) causes meningitis, necrotizing enterocolitis, sepsis, and bacteremia in neonates and children and has a high mortality rate. For rapid E. sakazakii detection, various differential and selective media containing α-glucosidase substrates, such as 5-bromo-4-chloro-3-indolyl-α-D-glucopyranoside (BCIG) or 4-methylumbelliferyl-α-D-glucoside (α-MUG), have been developed as only E. sakazakii exhibits α-glucosidase activity in the genus Enterobacter. However, Escherichia vulneris (family: Enterobacteriaceae) can also utilize α-glucosidase substrates, thereby resulting in false positives. Various iron sources are known to promote the growth of gram-negative bacteria. This study aimed to develop a selective medium containing α-glucosidase substrates for E. sakazakii detection that would eliminate false positives, such as those of E. vulneris, and to determine the role of iron source in the medium. Three previously developed (TPD) media, i.e., Oxoid, OK, and VRBG, and the medium developed in this study, i.e., NGTE, were evaluated using 58 E. sakazakii and 5 non-E. sakazakii strains. Fifty-four E. sakazakii strains appeared as fluorescent or chromogenic colonies on all four media that were assessed. Two strains showed colonies on NGTE medium and not on TPD media. In contrast, the remaining two strains showed colonies on TPD media and not on NGTE medium. None of the non-E. sakazakii strains showed fluorescent or chromogenic colonies on any of the evaluated media except E. vulneris, which showed colonies on TPD media and not on NGTE medium. This study demonstrated that the newly developed NGTE medium was not only equally efficient in promoting the growth of bacterial colonies when compared with the currently available media but also eliminated false positives, such as E. vulneris.

Microfluidic immunoassay using superparamagnetic nanoparticles in an enhanced magnetic field gradient (강화된 자기장 구배 하에서 나노자성입자를 이용한 미세유체 기반의 면역 측정)

  • Hahn, Young-Ki;Kang, Joo-H.;Kim, Kyu-Sung;Park, Je-Kyun
    • Journal of Sensor Science and Technology
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    • v.15 no.3
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    • pp.158-163
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    • 2006
  • This paper reports a novel immunoassay method using superparamagnetic nanoparticles and an enhanced magnetic field gradient for the detection of protein in a microfluidic device. We use superparamagnetic nanoparticles as a label and fluorescent polystyrene beads as a solid support. Based on this platform, magnetic force-based microfluidic immunoassay is successfully applied to analyze the concentration of IgG as model analytes. In addition, we present ferromagnetic microstructure connected with a permanent magnet to increase magnetic flux density gradient (dB/dx, ${\sim}10^{4}$ T/m), which makes limit of detection reduced. The detection limit is reduced to about 1 pg/mL.

"Turn-on" type colorimetric/fluorimetric probe for selective detection of Cu2+ at neutral pH condition

  • Lee, Hyun Jung;Saleem, Muhammad;Lee, Ki Hwan
    • Rapid Communication in Photoscience
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    • v.4 no.4
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    • pp.88-90
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    • 2015
  • The design and development of fluorescent chemosensors have recently been intensively explored for sensitive and specific detection of environmentally and biologically relevant metal ions in aqueous solution and living cells. Herein, we report the photophysical results of rhodamine B based fluorogenic and chromogenic receptor for selective copper detection in the complete organic or mixed aqueous-organic media at neutral pH under ambient condition. The ligand exhibited the remarkable increment in the fluorescence emission and UV-visible absorption signal intensities at 587 and 547 nm, respectively, on induction of copper ion while the ligand solution remain completely silent on addition of varieties of other metal ions.

In Situ Fluorescence Optical Detection Using a Digital Micromirror Device (DMD) for 3D Cell-based Assays

  • Choi, Jong-Ryul;Kim, Kyujung;Kim, Donghyun
    • Journal of the Optical Society of Korea
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    • v.16 no.1
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    • pp.42-46
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    • 2012
  • We have developed a fluorescence optical detection system using a digital micromirror device (DMD) for monitoring 3D cell culture matrices in situ. Full 3D imaging with fast scanning speed was implemented by the combined action of a DMD and a motorized stage. Imaging results with fluorescent microbeads measure the minimum axial resolution of the system as $6.3{\mu}m$, while full 1-mm scanning through 3D alginate-based matrix was demonstrated. For cell imaging, improved images were obtained by removing background fluorescence although the scanning distance was reduced because of low intracellular fluorescence efficiency. The system is expected to be useful to study various dynamics and behaviors of 3-dimensionally cultured cells in microfluidic systems.