• 제목/요약/키워드: Fluorescence detection system

검색결과 133건 처리시간 0.028초

Real-Time Detection of DNA Hybridization Assay by Using Evanescent Field Microscopy

  • Kim, Do-Kyun;Choi, Yong-Sung;Murakami, Yuji;Tamiya, Eiichi;Kwon, Young-Soo
    • KIEE International Transactions on Electrophysics and Applications
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    • 제11C권3호
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    • pp.85-90
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    • 2001
  • The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and other application area. So, the improvement of DNA detection system is very important for the determination of this hybridization reaction. In this study, we report the characterization of the probe and target oligonucleotide hybridization reaction using the evanescent field microscopy. First, we have fabricated DNA chip microarray. The particles which were immobilized oligonucleotides were arranged by the random fluidic self-assembly on the pattern chips, using hydrophobic interaction. Second, we have detected DNA hybridization reaction using evanescent field microscopy. The 5'-biotinylated probe oligonucleotides were immobilized on the surface of DNA chip microarray and the hybridization reaction with the Rhodamine conjugated target oligonucleotide was excited fluorescence generated on the evanescent field microscopy. In the foundation of this result, we could be employed as the basis of a probe olidonucleotide, capable of detecting the target oligonucleotide and monitoring it in a large analyte concentration range and various mismatching condition.

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경 X선 형광분석을 위한 모세관 광학소자 제작 (Fabrication of X-ray monocapillary optics for soft X-ray fluorescence analysis)

  • 조형욱;박병훈;김용민;최철희;최성희;김기홍;천권수
    • 한국방사선학회논문지
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    • 제5권6호
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    • pp.409-413
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    • 2011
  • 경 X선 형광분석 기법에 사용되는 X선 튜브는 X선의 휘도가 낮아 분석의 정밀도가 떨어지고 분석 시간 또한 오래 걸린다. 모세관 집광소자를 이용하면 X선 휘도의 이득(gain)을 최소 10 이상 얻을 수가 있다. 모세관 광학소자는 8.4keV의 텅스텐 특성방사선을 효율적으로 집광할 수 있도록 설계되었다. 파이렉스 유리로 된 모세관 모재를 풀러(puller)를 이용하여 45 g의 추에 $650^{\circ}$의 온도를 가하여 모세관 광학소자를 제작하였다. 모세관 광학소자의 제작은 총 460분이 소요되었으며 제작된 모세관 광학소자의 길이는 87 mm, X선 입사부의 직경은 300 ${\mu}m$, 출구부의 직경은 192 ${\mu}m$로 제작되었다. 제작된 모세관 광학소자를 경 X선 형광분석에 적용하면 황(S)과 같은 경원소 검출의 정밀도를 높일 수 있을 것이다.

3중 DNA probe를 이용한 FISH(fluorescence in situ hybridization) 기법으로 방사선에 의한 염색체 이상 분석 (Analysis of Chromosome aberrations by fluorescence in situ hybridization using triple chromosome-specific probes in human lymphocyte exposed to radiation)

  • 정해원;김수영;하성환
    • Journal of Radiation Protection and Research
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    • 제24권1호
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    • pp.45-53
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    • 1999
  • 각 염색체에 특이한 DNA probe를 이용하는 FISH기법은 방사선에 의해 유발된 상호전좌 및 삽입 등의 염색체의 구조적 변화를 측정하는 매우 효과적인 방법으로서 그 활용성이 증가되고 있다. 본연구는 방사선 피폭시 생물학적 선량측정법으로서 FISH기법을 활용하기 위하여 사람의 1, 2, 4번 염색체에 특이한 probe를 이용하여 고선량 단일 피폭시 유발된 각종 염색체 이상빈도를 관찰하고 이를 PAINT분류체계에 의해 분석하였다. 방사선 조사에 의한 염색체 이상빈도는 상호전좌(t)와 이동원염색체(dic)의 수가 선량 증가에 따라 같이 증가하는 것을 알 수 있으며 color junction의 수도 선량에 따라 증가하는 것을 알 수 있었다. 상호전좌의 빈도는 이동원 염색체의 빈도보다 상대적으로 높게 나타났다. 삽입(ins), 무동원염색체(ace), 및 환상염색체(r)의 수도 선량 증가에 따라 같이 증가하는 것을 알 수 있었다. 기존의 염색체재배열 분석방법과 비교해 볼 때 FISH기법은 다양한 형태의 염색체재해열을 보다 쉽게 관찰할 수 있게 하며 생물학적 선량제로서 중요한 역할을 할 것이라 기대된다.

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Automatic Reading System for On-off Type DNA Chip

  • Ryu, Mun-Ho;Kim, Jong-Dae;Kim, Jong-Won
    • Journal of Information Processing Systems
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    • 제2권3호
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    • pp.189-193
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    • 2006
  • In this study we propose an automatic reading system for diagnostic DNA chips. We define a general specification for an automatic reading system and propose a possible implementation method. The proposed system performs the whole reading process automatically without any user intervention, covering image acquisition, image analysis, and report generation. We applied the system for the automatic report generation of a commercialized DNA chip for cervical cancer detection. The fluorescence image of the hybridization result was acquired with a $GenePix^{TM}$ scanner using its library running in HTML pages. The processing of the acquired image and the report generation were executed by a component object module programmed with Microsoft Visual C++ 6.0. To generate the report document, we made an HWP 2002 document template with marker strings that were supposed to be searched and replaced with the corresponding information such as patient information and diagnosis results. The proposed system generates the report document by reading the template and changing the marker strings with the resultant contents. The system is expected to facilitate the usage of a diagnostic DNA chip for mass screening by the automation of a conventional manual reading process, shortening its processing time, and quantifying the reading criteria.

Microfluidic LOC 시스템 (Microfluidic LOC System)

  • 김현기;구홍모;이양두;이상렬;윤영수;주병권
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2004년도 하계학술대회 논문집 Vol.5 No.2
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    • pp.906-911
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    • 2004
  • In this paper, we used only PR as etching mask, while it used usually Cr/AU as etching mask, and in order to fabricate a photosensor has the increased sensitivity, we investigated on the sensitivity of general type and p-i-n type diode. we designed microchannel size width max 10um, min 5um depth max 10um, reservoir size max 100um, min 2mm. Fabrication of microfluidic devices in glass substrate by glass wet etching methods and glass to glass fusion bonding. The p-i-n diode has higher sensitivity than photodiode. Considering these results, we fabricated p-i-n diodes on the high resistive($4k{\Omega}{\cdot}cm$) wafer into rectangle and finger pattern and compared internal resistance of each pattern. The internal resistance of p-i-n diode can be decreased by the application of finger pattern has parallel resistance structure from $571\Omega$ to $393\Omega$.

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A Spectral Comparison Study of PDT Drugs - ALA and ALA-Hexyl ester

  • Yao, Chen-Ji;Peng Qian;JodI H-J
    • Journal of Photoscience
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    • 제9권2호
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    • pp.521-523
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    • 2002
  • 5-aminolevulinic acid (ALA) has been used to stimulate endogenous protoporphyrin IX (PpIX) in tumor and then initiate PDT. Recently, ALA-Hexyl ester (He-ALA) was found much effective than ALA on producing PpIX in cancer cells. To clarify the transportation mechanism of ALA and He-ALA, the detection of them is the important step. ALA and its derivatives all don't emit fluorescence, so the Raman spectroscopy was used here for the direct detection of ALA and He-ALA. The results showed that ALA and He-ALA have the common strong Raman peaks at 2930, 2950 CM$\^$-1/, due to the CH$_2$ vibration. The peak 3050 CM$\^$-1/ appeared in ALA spectrum can be attributed to OH vibration, while the peaks of 2860, 2900 CM$\^$-1/ in He-ALA spectrum were assigned as the modes of CH$_3$. This Raman spectral characteristic is consistence with the structure difference of He-ALA and ALA. Thus, Raman spectroscopy provides a new way to detect and distinguish ALA and He-ALA, and could be explored further in biology system.

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DNA Hybridization 검출 센서를 이용한 매치 및 미스매치 DNA hybridization 특성 연구 (A Study on Match and Mismatch DNA Hybridization properties Using DNA Hybridization Detection Sensor)

  • 김도균;권영수
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 2003년도 추계학술대회 논문집 전기물성,응용부문
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    • pp.89-91
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    • 2003
  • The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and other application area. So, the improvement of DNA detection system is very important for the determination of this hybridization reaction. In this study, we report the characterization of the probe and target oligonucleotide hybridization reaction using the evanescent field microscopy. First, we have fabricated DNA chip microarray. The particles which were immobilized oligonucleotides were arranged by the random fluidic self-assembly on the pattern chips, using hydrophobic interaction. Second, we have detected DNA hybridization reaction using evanescent field microscopy. The 5'-biotinylated probe oligonucleotides were immobilized on the surface of DNA chip microarray and the hybridization reaction with the Rhodamine conjugated target oligonucleotide was excited fluorescence generated on the evanescent field microscopy. In the foundation of this result, we could be employed as the basis of a probe olidonucleotide, capable of detecting the target oligonucleotide and monitoring it in a large analyte concentration range and various mismatching condition.

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FMOC 표식에 의한 Sugar Chain의 분석 (Application of FMOC-Cl for the Quantitative Determination of N-linked Oligosaccharides)

  • 김동현;황보식;정구용
    • 한국식품과학회지
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    • 제29권4호
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    • pp.630-634
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    • 1997
  • N-glycanase처리에 의해 얻어진 1-amino-oligosaccharide를 FMOC-Cl로 표식 하는 방법을 확립하였다. 이 방법에 의해 얻어진 FMOC-표식 oligosaccharides는 기존의 방법보다 약 4배의 감도를 나타내었다. Amido 80 column을 사용한 분석에 의하여, Man 5-9 GlcNAc 2 amines의 5개의 성분은 각각 분리되어 용출 되었으며, 회수율은 기존의 방법과 거의 같았다. 1-amino-oligosaccharides는 0.05 pmol에서 1.5 pmol 사이에서 직선적인 관계를 나타내었으며, 2-aminopyridine에 의한 표식과 비교하였을 때, 안정된 화합물을 형성하고 있으며, 감도가 높아 미량의 oligosaccharides의 분석에 매우 적합한 것이 확인되었다.

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식품병마개 PVC gasket과 식품에 함유된 semicarbazide의 분석 (Determination of Semicarbazide in PVC Gaskets of Food Bottle Cap and Foods)

  • 박상욱;이광호;곽인신;전대훈;이시경
    • 한국식품과학회지
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    • 제37권3호
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    • pp.334-338
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    • 2005
  • 본 연구는 식품병마개 PVC 가스켓과 식품에 함유된 SEM을 효과적으로 분석하기 위해 연구했다. SEM은 발포제로 사용되는 ADC의 열분해산물로 알려졌다. 그러나 SEM은 분자량이 낮으며 ultraviolet light나 fluorescence에 활성을 갖지 않는다. 따라서 2-NBA-SEM으로 유도체화하여 HPLC triple column system으로 분석했을 때 상관관계는 0.9997이상이며 검출한계는 0.48ng/g으로 나타났다. 식품병마개 PVC 가스켓에서 SEM의 검출율은 77.08%로 나타났으며, 유통중인 병마개 PVC 가스켓에서 SEM은 812.20-5771.30ng/g 수준이였다. 그리고 회수율은 PVC 가스켓과 식품에서 각각 92.12-98.71%와 83.45-97.33%로 측정되었다.

Gallic acid가 Lipopolysaccharide로 활성화된 마우스 대식세포의 케모카인과 성장인자 생성에 미치는 영향 (Inhibitory Effect of Gallic acid on Production of Chemokine and Growth Factor in Mouse Macrophage Stimulated by Lipopolysaccharide)

  • 박완수
    • 동의생리병리학회지
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    • 제24권4호
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    • pp.586-591
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    • 2010
  • Chemokine and Growth Factor are major mediumtors of immuno-inflammatory pathway. The purpose of this study is to investigate whether productions of Chemokine and Growth Factor in lipopolysaccharide (LPS)-induced mouse macrophage RAW 264.7 cells are modulated by Gallic acid (GA), which is easily founded in tannin-containing natural materials such as red wine, green tea, grape juice, and Corni Fructus. Productions of Chemokine and Growth Factor were analyzed by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. At first, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, the antibody-conjugated beads were added and incubated for 30 minutes. After incubation, detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System. Based on fluorescence intensity, concentrations of Chemokine and Growth Factor were determined. The results of the experiment are as follows. GA significantly inhibited the production of interferon-inducible protein (IP)-10, keratinocyte-derived chemokine(KC), and vascular endothelial growth factor (VEGF) in LPS-induced RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). GA significantly inhibited the production of monocyte chemoattractant protein-1(MCP-1) and macrophage-colony stimulating factor(M-CSF) in LPS-induced RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). GA diminished the production of granulocyte macrophage-colony stimulating factor (GM-CSF) in LPS-induced RAW 264.7 cells. But GA did not show the inhibitory effect on the production of leukemia inhibitory factor (LIP) and macrophage inflammatory protein (MIP)-2 in LPS-induced RAW 264.7 cells. These results suggest that GA has the immuno-modulating activity related with its inhibitory effects on the production of IP-10, KC, MCP-1, VEGF, and M-CSF in LPS-induced macrophages.