• Title/Summary/Keyword: fluorometer

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Evaluation of the ETRmax in Microalgae Using the PHYTO-PAM Fluorometer (광합성 측정기를 이용한 미세조류의 광합성 효율 측정)

  • Cho, Eun-Seob;Lee, Pil-Yong;Oh, Hyun-Ju;Choi, Yoon-Seok;Choi, Yang-Ho;Lee, Sam-Geun
    • Journal of Environmental Science International
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    • v.15 no.8
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    • pp.727-735
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    • 2006
  • In this study, the PHYTO-PAM-fluorometric method was used to evaluate the ETR$_{max}$ in terms of sensitivity to DIN/DIP against 14 microalgae: Prorocentrum micans, Heterocapsa triquetra, Gymnodinium impudicum, Cymnodinium catenatum, Amphidinium caterae, Chlorella vulgaris, Chroococcus minutus, Microcystis aeruginosa, Chlorella ellipsoidea, Nannochloris oculata, Oocystis lacustris, Chroomonas salina, Gloeocystis gigas, and Prymnessium parvum. We found that P. micans, H. triquetra, and A. caterae exposed to the maximum level of DIN/DIP were significantly smaller in the ETR$_{max}$ than that of the minimum and moderate mixture. Unlikely the ETR$_{max}$, the initial slope alpha was not significantly different at the level of 60 DIN/DIP. In G. catenatum, the moderate levels of 15 and 20 in DIN/DIP were found to be significantly different from the ETR$_{max}$ at Chl-Ch4. Gymnodinium impudicum had a higher value than that of the ETR$_{max}$ than that of dinoflagellates used in this study, ranging from 306.1 (Ch4, DIN/DIP: 10) to 520.1 (Ch4, DIN/DIP: 30). The ETR$_{max}$ value obtained from other microalgae was similar to C. impudicum at any of the ratios of DIN/DIP and channels. Consequently, the influence of offshore water current assures us of the suppression of photosynthesis and electron transport rate in dinoflagellates. Gymnodinium impudicum has not been researched in the area of red tides in Korea, but it will be enough to creat the massive algal blooms in the future because of higher potential photochemical availability.

The Early Detection of the Spore Using Sonication and Fluorescent Dye in the Field (현장에서 초음파 파쇄와 형광시약을 이용한 포자의 조기 탐지)

  • Ha, Yeon-Chul;Choi, Ki-Bong
    • KSBB Journal
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    • v.26 no.4
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    • pp.305-310
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    • 2011
  • This study was carried out to establish the optimum condition of cell disruption with a sonicator for the detection of the spore, Bacillus anthracis ${\Delta}$-sterne for the purpose of developing automatic fluorometer. The efficiency of sonication on the ${\Delta}$-sterne spore disruption was very weak. The ${\Delta}$-sterne spore with zirconia bead showed greater disruption than the ${\Delta}$-sterne spore alone when sonificated. The volumn of the zirconia bead added in the spore solution has little effect on the disruption efficiency. The detection limit of the ${\Delta}$-sterne spore with zirconia bead and the ${\Delta}$-sterne spore alone was $10^6$ CFU/mL and $5{\times}10^7$ CFU/mL respectively, when sample was sonicated for 20 seconds with a sonicator probe of 13 mm diameter.

Time-resolved Fluoroimmunoassay for the Measurement of 17$\beta$-Estradiol using Anti-idiotypic Antibody (Anti-idiotype 항체를 이용한 17$\beta$-Estradiol 측정을 위한 Time-resolved Fluoroimmunoassay)

  • 김윤규;김창규;박성민;이치호;이원창;최영숙;김종배
    • Korean Journal of Animal Reproduction
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    • v.16 no.4
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    • pp.325-333
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    • 1993
  • A competitive type immunoassay method for 17$\beta$-estradiol(E2) based on the idiotypic anti-idiotypic antibody and time-resolved fluorescence is described. The anti-idiotypic antibody(Ab2) produced to E2 binding site of the primary idiotype antibody (Ab1) was labelled with europium and was allowed to compete with E2 standards or serum sample for the binding sites of Ab1 which was bound to 2nd antibody captured ontothe surface of microtitre plates. Fluorescence measured by time-resolved fluorometer was inversely proportional to the concentration of E2 over the range 5~500pg/well. The sensitivity of the assay was 5pg per well which was compatible with that ofradioimmunoassay using the same Ab1 and 3H-E2 as a tracer. One great advantage of this method described here was to enable antibodies to be labelled instead of haptens, and thus makes it easier to develop sensitive and robust immunoassay systems specially for haptens.

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Characteristics of Trichloroethene and Tetrachloroethene Sensing Optical Fiber Biosensor Using Toluene-o-monooxygenase and Fluoresceinamine (Toluene-o-monooxygenase와 Fluoresceinamine을 이용한 Trichloroethene와 Tetrachloroethene 감지용 광섬유 바이오센서의 특성)

  • Ryoo, Doohyun
    • Journal of Soil and Groundwater Environment
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    • v.23 no.4
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    • pp.42-47
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    • 2018
  • E. coli TG1 pBS TOM Green was cultured to produce toluene-o-monooxygenase (TOM). A biosensor system was successfully constructed using purified TOM to effectively detect trichloroethene (TCE) and tetrachloroethene (PCE), which represent some of the major contaminants in groundwater and soil. In order to utilize TOM as a sensor, NADH, a biological oxidizer, was replaced with hydrogen peroxide which is a chemical oxidizing agent. A three-layered sandwich-type sensing tip was fabricated on the outside of the hydrophilic polyvinylidene fluoride membrane. TCE and PCE were applied to the sensor and the hydrogen ions were measured by a fiber optic fluorometer using fluoresceinamine. Calibration curves were obtained for TCE and PCE in the concentration range of 0.2-100 mg/l, and the detection limit of the system was $10{\mu}g/l$ for TCE and PCE.

Bipolar Charge Distribution of Nano Particles Passing through the Dielectric Barrier Discharge Reactor (DBD(Dielectric Barrier Discharge)에 의해 하전된 나노입자의 양극성 대전량 분포)

  • Ji, Jun-Ho;Kang, Suk-Hoon;Byeon, Jung-Hoon;Hwang, Jung-Ho
    • Proceedings of the KSME Conference
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    • 2003.04a
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    • pp.1684-1689
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    • 2003
  • Dielectric Barrier Discharges (DBD) in oxygen and air are well established for the production of large quantities of ozone and are more recently being applied to a wider range of after treatment processes for HAPs(Hazardous Air Pollutants). The potential use as a charger for particle collection are not well known. In this work, we measured charge distribution of nanometer or submicron sized particles passing through the dielectric barrier discharge reactor. The bipolar charge characteristics of particles passing DBD reactor were investigated. Fluorometric method using uranine particles and a fluorometer was employed to examine the bipolar charging characteristics of the charged particles by DBD reactor. Finally, the charge distributions of particles were determined from the electrical mobility classification using DMA.

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Zooplankton Biomass and Size Estimation Using a Multi-frequency Acoustic System (고주파 다주파 음향시스템을 이용한 동물성 플랑크톤의 크기별 생물량 추정)

  • Hwang, Bo-Kyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.41 no.1
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    • pp.54-60
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    • 2008
  • High- and multi-frequency acoustic systems can measure a zooplankton patch successively and estimate the spatial distribution and abundance of zooplankton according to size using a multi-frequency inversion (MFI) method. This study measured zooplankton distribution to a depth of 150m using a multi-frequency acoustic system (TAPS-6), installed on a CTD system with a fluorometer and analyzed it using the MFI method. Simultaneously, zooplankton samples were collected by north pacific standard (NORPAC) net to confirm the species composition. The results showed that the combined method is valuable for estimating the zooplankton profile in detail and investigating the relationship between the zooplankton and phytoplankton profiles.

Miniaturized Fluorometer Based on Total Internal Reflector and Condensing Mirror

  • Jang, Dae-Ho;Yoo, Jae-Chern
    • Journal of the Optical Society of Korea
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    • v.17 no.1
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    • pp.81-85
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    • 2013
  • A miniaturized fluorescence detection system based on total internal reflection (TIR) configuration, which is applicable to detecting the presence of biological materials labeled with fluorescence dye in micro total analysis systems (${\mu}TAS$), is proposed. In conventional fluorescence testing and analysis devices, interference between the excitation light beam and the emitted light from dyes is unavoidable. This paper presents a fluorescence detection system based on TIR configuration that allows the excitation light beam and the emitted light to be spatially perpendicular to each other so as to minimize the interference where fluorescence emission is detected at the orthogonal angle to the excitation beam. We achieved the limit of detection of about 5 nmol/L with a high linearity of 0.994 over a wide range of 6-FAM mol concentration, being comparable to that in earlier studies.

Photosynthetic Responses of the Benthic Diatom Nitzschia sp. to Selected Heavy Metals and Herbicides (일부 중금속과 제초제에 대한 저서규조류 Nitzschia sp.의 광합성 반응)

  • Kang, Eun-Ju;Choi, Tae-Seob;Kim, Kwang-Young
    • ALGAE
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    • v.22 no.4
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    • pp.319-323
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    • 2007
  • This study was conducted with using chlorophyll a fluorescence (indicated as photosynthetic activity) to examine the toxic effect of 96 h exposure of heavy metals and herbicides on the benthic diatom Nitzschia sp. which was isolated from pristine sediment in Pamquat Harbour, Nova Scotia, Canada. Samples of benthic diatom were exposed to 0, 0.01, 0.1 and 1 mg L–1 of copper, 0, 1, 10 and 100 mg L–1 of chrome (VI), 0, 2.45, 24.5 and 245 mg L–1 of paraquat dichloride, and 0, 4.37, 43.7 and 437 mg L–1 of alachlor during 96 hours. The effective quantum yield of photochemistry (ΔF/Fm’) was evaluated by subjecting light acclimated samples to saturating pulses of light using a pulse amplitude modulated (PAM) fluorometer. The impact of heavy metals on Nitzschia sp. photosynthesis was not severe in < 1 mg L–1 but in the high concentrations (> 1 mg L–1) clearly increased toxic stress during 96 h. Herbicides had a limited impact during the exposure period but clearly increased stress on the benthic diatom with increasing concentrations. Acute response of Nitzschia sp. to selected heavy metals and herbicides was characterized, and the capacity of a benthic diatom to tolerate and recover from toxic stress was assessed.

Assay System for N-acylethanolamines Degradation Enzyme, N-acylethanolamine-hydrolyzing Acid Amidase

  • Kim, Dae-Woong;Kim, Gun-Joong;Kim, Hae-Jo;Ghil, Sung-Ho
    • Biomedical Science Letters
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    • v.18 no.4
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    • pp.438-444
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    • 2012
  • N-acylethanolamines (NAEs) including endocannabinoids, anadamide, are long chain fatty acid ethanolamines and express ubiquitously in animal and plant tissues. NAEs have several pharmacological effects including anti-inflammatory, analgesic and anorexic effects. The levels of NAEs in tissues are strictly regulated by synthesizing and hydrolyzing enzymes because NAEs are not stored in the cell but rather made on demand. NAEs are hydrolyzed to free fatty acids and ethanolamines by fatty acid amide hydrolase and N-acylethanolamine-hydrolyzing acid amidase (NAAA). Here, we suggest the fluorescence-based assay system for NAAA. We developed N-(4-methy-2-oxo-2H-chromen-7-yl)palmitamide (PAAC) as a fluorogenic substrate for NAAA and we also generated NAAA stably expressing COSM6 cell line. When extracts of cells expressing NAAA were incubated with PAAC, NAAA specifically hydrolyzed PAAC to palmitic acids and fluorogenic dye, coumarin. Release of coumarin was monitored by using fluorometer. NAAA hydrolyzed PAAC with an apparent Km of $20.05{\mu}M$ and Vmax of 32.18 pmol/mg protein/min. This assay system can be used to develop inhibitors or activators of NAAA.

Live Cell Detection of Monoclonal Antibody Light and Heavy Chain mRNAs using Molecular Beacons (분자 비컨을 이용한 살아 있는 세포에서 단일클론항체 경쇄와 중쇄 mRNA 검출에 의한 세포주 선별방법)

  • Jeong, Seunga;Rhee, Won Jong
    • KSBB Journal
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    • v.31 no.1
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    • pp.33-39
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    • 2016
  • Developing the method for the selection of animal cell line producing therapeutic monoclonal antibody (mAb) is invaluable as its market is rapidly growing. Although the quality of produced mAb is as important as quantity, however there is no method developed for the selective screening of cell lines on the basis of both quantity and quality. From recent reports, the ratio of light and heavy chain mRNAs of mAb in the cell is a key parameter for the indication of product quality. Therefore, it is obvious that developing the novel method that can detect both light and heavy chain mRNAs in single live cell will provide unprecedented opportunities in bio-industry. Here, we have constructed oligonucleotide probes, molecular beacons for the detection of light or heavy chain mRNAs, respectively, in the live cells producing mAbs. Both beacons showed increased fluorescent intensity after transient transfection of plasmid expressing mAbs analyzed by fluorometer. Flow cytometric analysis clearly demonstrated that both molecular beacons can simultaneously detect the expression of light and heavy chain mRNAs of mAb in the same cell. The technique described in the thesis provides the new direction and concept for developing the method for the smart selection of cell lines producing recombinant proteins including therapeutic mAbs.