• Title/Summary/Keyword: fluorescent detection

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The Early Detection of the Protein Toxin using Sanification and Fluorescent Dye in the Field (현장에서 초음파 파쇄와 형광시약을 이용한 단백질 독소의 조기 탐지)

  • Ha, Yeon-Chul;Choi, Ki-Bong;Kim, Seong-Joo;Choi, Jung-Do
    • KSBB Journal
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    • v.22 no.1
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    • pp.48-52
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    • 2007
  • This study was carried out to establish the optimum disruption condition of a sonificator for the protein toxin for the purpose of developing automatic biological agent detector equipped a sonificator. One of the best-known collisional quenchers is molecular oxygen, which quenches almost all known fluorophores. The sonification does an excellent job of degassing, which decreased the quenching effect and increased the fluorescence quantity. The fluorescence measurement for the protein using 0.7 X fluorescent dye concentration and above must be done in 1 minute and the fluorescence measurement for the protein using 0.3 X fluorescent dye concentration and below has to be done between 2 and 3 minute. The fluorescence quantity of the sonificatied protein sample was much higher that of the non-sonificatied protein sample. Sonificating the sample turned out to be favorable for the fluorescence measurement when measuring at the low protein concentration.

Cloning of Transgenic Rabbit Embryos Expressing Green Fluorescent Protein Gene by nuclear Transplantation (Green Fluorescent Protein 발현 토끼 수정란의 핵이식에 의한 복제)

  • Kang, T. Y.;Yin, X. J.;Rho, G. J.;Lee, H.;Chae, Y. J.;Lee, H. J
    • Journal of Embryo Transfer
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    • v.15 no.2
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    • pp.167-173
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    • 2000
  • The principal objective of this study was to clone transgenic embryos in order to improve the efficiency of transgenic animal production by the combination of microinjection and nuclear transplantation techniques. Mature female New Zealand White rabbits were superovulated by eCG and hCG treatments, fllowed by natural mating. Zygotes were collected from the oviducts at 18∼22 h after hCG injection by flushing with D-PBS containing 5% fetal calf serum(FCS). Two to three picoliters of green fluorescent protein(GFP) gene wa microinjected into male pronucleus. The foreign gene-injected zygotes were cultured in TCM-199 or RD medium containing 10% FCS with a monolayer of rabbit oviductal epithelial cells in a 5% CO2 incubator. The morulae expressing GFP gene were selected and their blastomeres were separated for the use of nuclear donor. Following nuclear transplantation of fluorescence-positive morula stage blastomeres, 13 (21.3%) out of 61 fused oocytes developed to blastocyst stage and all of the cloned blastocysts expressed GFP. The results indicate that the screening of transgene in rabbit embryos by GFP detection could be a promisible method for the preselection of transgenic embryos. Also the cloning of preselected transgenic embryos by nuclear transplantatin could be efficiently applied to the multiple production of transgenic animals.

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Detection of Vibrio alginolyticus in Penaeus chinensis Using an Indirect fluorescent Antibody Technique

  • Heo, Moon-Soon;Lee, Sang-Hyeon;Son, Hong-Joo;Choon, Bok-Song;Lee, Je-Hee;Yeo, In-Okyy;Lee, Jung-Jae
    • Journal of Life Science
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    • v.10 no.1
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    • pp.24-27
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    • 2000
  • For the rapid diagnosis of vibriosis in penaeid shirmp, the indirect fluorescent antibody technique(IFAT) was established to detect Vibrio alginolyticus. The titers of the antisera used for this experiment were above 1280. Vibrio alginolyticus possesses the specific antigen, and also have antigens shared with other strains. When an V. alginolyticus-infected adult shirmp was tested by IFAT, V. alginolyticus was detected mainly in the muscle tissues near the injection point and the haemolymph but only few in other tissues. This result indicates that the pathogen bacteria could be detected by IFAT. Thus, it is suggested IFAT is more convenient and sensitive method than conventional plate method for the diagnosis of induced Vibrio infection in the penaeid shrimps.

The analysis of impurities in rare earth oxide for fluorescent substance by ICP-MS (ICP-MS에 의한 형광체용 $(Y,\;Eu)_2O_3$ 중 불순물 분석 연구)

  • Kim, Sang-Kyoung;Jang, Seung-Kyu
    • Analytical Science and Technology
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    • v.7 no.1
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    • pp.1-10
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    • 1994
  • A survey was made on detection limit, reproducibility, matrix effect, linear dynamic range and the memory effect of yttrium and europium in order to analyze rare earth elements which exist as impurities in the rare earth oxide which is raw materials of fluorescent substance. When analysing a certain amount of thulium quantitatively using inductively coupled plasma mass spectrometry, it was found that the analysis was interfered with $EuO^+$ which is one of polyatomic ions caused by plasma. As the intensity of thulium linearly proportional to the europium concentration, it was possible to the determine the actual concentration of thulium.

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Optical Tracking of Three-Dimensional Brownian Motion of Nanoparticles

  • Choi C. K.;Kihm K.D.
    • Journal of the Korean Society of Visualization
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    • v.3 no.1
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    • pp.3-19
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    • 2005
  • Novel optical techniques are presented for three-dimensional tracking of nanoparticles; Optical Serial Sectioning Microscopy (OSSM) and Ratiometric Total Internal Reflection Fluorescent Microscopy (R-TIRFM). OSSM measures optically diffracted particle images, the so-called Point Spread Function (PSF), and dotermines the defocusing or line-of-sight location of the imaged particle measured from the focal plane. The line-of-sight Brownian motion detection using the OSSM technique is proposed in lieu of the more cumbersome two-dimensional Brownian motion tracking on the imaging plane as a potentially more effective tool to nonintrusively map the temperature fields for nanoparticle suspension fluids. On the other hand, R-TIRFM is presented to experimentally examine the classic theory on the near-wall hindered Brownian diffusive motion. An evanescent wave field from the total internal reflection of a 488-nm bandwidth of an argon-ion laser is used to provide a thin illumination field of an order of a few hundred nanometers from the wall. The experimental results show good agreement with the lateral hindrance theory, but show discrepancies from the normal hindrance theory. It is conjectured that the discrepancies can be attributed to the additional hindering effects, including electrostatic and electro-osmotic interactions between the negatively charged tracer particles and the glass surface.

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Application of Micro-Thin Laser sheet and Mixed Solvent for Micro-LIF Measurement in a Microchannel (마이크로 채널 내부의 Micro-LIF 측정을 위한 마이크로 레이저 평면빔과 혼합용매의 적용)

  • Yoon Sang Youl;Kim Jae Min;Kim Kyung Chun
    • 한국가시화정보학회:학술대회논문집
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    • 2004.11a
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    • pp.86-89
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    • 2004
  • One most feasible 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 successfully achieved so far due to various limitations in the light illumination and fluorescence signal detection. The present study demonstrates a novel method to provide an ultra thin laser sheet beam having five(5) microns thickness by use of a micro focus laser line generator. The laser sheet beam illuminates an exact plane of concentration measurement field to increase the signal to noise ratio and considerably reduce the depth uncertainty. Nile Blue A was used as fluorescent dye for the present LIF measurement. The enhancement of the fluorescent intensity signals was performed by a solvent mixture of water $(95\%)$ and ethanol (EtOH)/methanol (MeOH) $(5\%)$ mixture. To reduce the rms errors resulted from the CCD electronic noise and other sources, an expansion of grid size was attempted from $1\times1$ to 3(3 or 5(5 pixel data windows and the pertinent signal-to-noise level has been noticeably increased accordingly.

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A Carbazole Based Bimodal "Turn-On" Fluorescent Probe for Biothiols (Cysteine/Homocysteine) and Fluoride: Sensing, Imaging and its Applications

  • Kaur, Matinder;Yoon, Byungkwon;Kumar, Rajesh;Cho, Min Ju;Kim, Hak Joong;Kim, Jong Seung;Choi, Dong Hoon
    • Bulletin of the Korean Chemical Society
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    • v.35 no.12
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    • pp.3437-3442
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    • 2014
  • A well-known carbazole-based precursor (probe 1) was used for the detection of cysteine/homocysteine and fluoride. Probe 1 shows a "turn-on" response to cysteine/homocysteine and fluoride via enhancement in emission intensity at 442 nm and 462 nm respectively, in solutions and living cells. Furthermore, probe 1 behaves as a fluorescent molecular switch between cysteine/homocysteine and fluoride as the chemical inputs, which have been used for the development of a combinatorial logic circuit and a molecular keypad lock.

A Simple Carbazole-based Schiff Base as Fluorescence "off-on" Probe for Highly Selective Recognition of Cu2+ in Aqueous Solution

  • Tang, Lijun;Wu, Di;Hou, Shuhua;Wen, Xin;Dai, Xin
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2326-2330
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    • 2014
  • A carbazole-based Schiff base CB2 was synthesized and applied as a highly selective and sensitive fluorescent probe for $Cu^{2+}$ in $H_2O$-DMSO (8/2, v/v, pH = 7.4) solution. CB2 exhibits an excellent selectivity to $Cu^{2+}$ over other examined metal ions with a prominent fluorescence "turn-on" at 475 nm. CB2 and $Cu^{2+}$ forms a 1:2 binding ratio complex with detection limit of $9.5{\mu}M$. In addition, the $Cu^{2+}$ recognition process is hardly interfered by other examined metal ions.

Spectrofluorimetric determination of EDTA with Cu(II)-tiron chelate (Cu(II)-tiron 킬레이트를 이용한 EDTA 분광형광법 정량)

  • Choi, Hee-Seon
    • Analytical Science and Technology
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    • v.24 no.3
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    • pp.231-235
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    • 2011
  • A spectrofluorimetric method for the determination of EDTA in real samples such as mayonnaise, powder detergent and cleansing cream with tiron (4,5-dihydroxy-1,3-benzenedisulfonic acid) as a fluorimetric reporter was developed. When tiron is chelated with Cu(II), the fluorescent intensity is decreased by a quenching effect. However, when Cu(II)-tiron chelate reacts with EDTA, fluorescent intensity is increased as tiron is released. Several experimental conditions such as pH of the sample solution, the amount of Cu(II), the amount of tiron, heating temperature and heating time were optimized. Fe(III) interfered more seriously than any other ions, interference of Fe(III) could be disregarded, because Fe(III) was scarcely contained in selected real samples. The linear range of EDTA was from $8.0{\times}106{-8}\;M$ to $2.0{\times}10^{-6}\;M$. With this proposed method, the detection limit of Fe(III) was $5.2{\times}10^{-8}\;M$. Recovery yields of 92.7~99.3% were obtained. Based on experimental results, it is proposed that this technique can be applied to the practical determination of EDTA.

The Rapid Differentiation of Toxic Alexandrium and Pseudo-nitzschia Species Using Fluorescent Lectin Probes

  • Cho, Eun-Seob;Park, Jong-Gyu;Kim, Hak-Gyoon;Kim, Chang-Hoon;Rhodes, Lesley L.;Chung, Chang-Soo
    • Journal of the korean society of oceanography
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    • v.34 no.3
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    • pp.167-171
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    • 1999
  • Since toxic Alexandrium catenella and non-toxic A. fraterculus are morphologically similar, they are difficult to discriminate under the light microscope. However, a novel technology, such as fluorescein isothiocyanate (FITC)-conjugated lectin probes enables easy and rapid differentiation. Toxic A. catenella bound seven different lectins, whereas the non-toxic A. fratercuzus did not bind Arachis hypogaea (PNA) lectin. In addition, Pseudo-nitrschia species in this study were also difficult to identify to species level with light microscope techniques, but it was possible to classify them using fluorescent lectins. Pseudo-nitzschia multistriata, P. subfraudulenta and P. pungens bound Canavalia ensiformis (ConA), whereas P. subpaclfica did not, and P. pungens also bound Ricinus communis (RCA). These results imply that lectin could be used as a critical tool in the differentiation of P. multistriata, P. subfraudulenta and P. pungens. However, P. subpacifica was not differentiated by the lectins tested. Therefore, it isconcluded that lectin probes are useful for discriminating toxic A. catenella from non-toxic A. fraterculus, and for the identification of some Pseudo-nitzschia species. In addition, this method has a great potential to speed and detection between non-toxic and toxic harmful algal blooms (HABs) in Korean biotoxin monitoring systems.

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