• 제목/요약/키워드: Fluorescence probes

검색결과 110건 처리시간 0.03초

Calibration-free real-time organic film thickness monitoring technique by reflected X-Ray fluorescence and compton scattering measurement

  • Park, Junghwan;Choi, Yong Suk;Kim, Junhyuck;Lee, Jeongmook;Kim, Tae Jun;Youn, Young-Sang;Lim, Sang Ho;Kim, Jong-Yun
    • Nuclear Engineering and Technology
    • /
    • 제53권4호
    • /
    • pp.1297-1303
    • /
    • 2021
  • Most thickness measurement techniques using X-ray radiation are unsuitable in field processes involving fast-moving organic films. Herein, we propose a Compton scattering X-ray radiation method, which probes the light elements in organic materials, and a new simple, non-destructive, and non-contact calibration-free real-time film thickness measurement technique by setting up a bench-top X-ray thickness measurement system simulating a field process dealing with thin flexible organic films. The use of X-ray fluorescence and Compton scattering X-ray radiation reflectance signals from films in close contact with a roller produced accurate thickness measurements. In a high-thickness range, the contribution of X-ray fluorescence is negligible, whereas that of Compton scattering is negligible in a low-thickness range. X-ray fluorescence and Compton scattering show good correlations with the organic film thickness (R2 = 0.997 and 0.999 for X-ray fluorescence and Compton scattering, respectively, in the thickness range 0-0.5 mm). Although the sensitivity of X-ray fluorescence is approximately 4.6 times higher than that of Compton scattering, Compton scattering signals are useful for thick films (e.g., thicker than ca. 1-5 mm under our present experiment conditions). Thus, successful calibration-free thickness monitoring is possible for fast-moving films, as demonstrated in our experiments.

방사선에 의한 염색체이상과 DNA 함량과의 관계 (Relationship between the DNA content of human chromosome and their contribution to radiation-induced chromosome aberration analysed by fluorescence in situ hybridization(FISH))

  • 정해원;김수영;하성환;김태환;조철구
    • Journal of Radiation Protection and Research
    • /
    • 제26권2호
    • /
    • pp.101-111
    • /
    • 2001
  • 본 연구는 염색체 1, 2, 4, 7, 8, 9 및 21번 염색체의 DNA probe를 이용하여 2Gy의 방사선을 조사한 후 DNA 양을 감안한 기대치와 관찰치의 차이를 비교함으로서 각 염색체의 방사선에 대한 감수성을 평가하여 궁극적으로 방사선 피폭시 생물학적 선량계로서 FISH기법의 타당성을 평가하고자 하였다. 1번 및 4번 염색체의 경우 상호전좌와 이동원 염색체의 관찰치가 기대치보다 더 높게 나타났으며 이와 반대로 2, 7, 8 및 9번 염색체의 경우 상호전좌와 이동원염색체의 관찰치 모두 기대치보다 낮게 나타났다. 2번 및 4번 염색체의 경우 1번 염색체보다 더 많은 acentric fragment의 빈도를 나타내었다. 1, 2, 및 4번 염색체 3종을 조합했을 때 상호전좌의 경우 관찰치와 기대치는 세포 100개당 25.5 및 25.40으로 차이가 없었으며 이동원염색체의 경우 13.25 및 13.2로 역시 거의 차이가 없게 나타났다. 따라서 본 연구결과 방사선 피폭시 발생하는 염색체이상 빈도는 염색체마다 DNA 양에 비례해 나타나지 않을 수 있어 각 염색체마다 방사선 감수성에 차이가 있을 것으로 판단된다. 또한 방사선 피폭시 생물학적 선량계로서 1, 2 및 4번을 동시에 관찰 할 경우 염색체 FISH 법을 활용하기 위하여 적절한 염색체 조합이라고 판단된다.

  • PDF

탄소 나노튜브와 생체 분자와의 결합을 통한 나노-바이오 응용 (Nano-Bio Applications Using Carbon Nanotube-Biomolecule Conjugates)

  • 황응수;조승범;홍상현;정혜진;차창용;최재붕;김영진;백승현
    • 한국정밀공학회지
    • /
    • 제23권3호
    • /
    • pp.179-186
    • /
    • 2006
  • Single-walled carbon nanotubes (SWNT) exhibit strong Raman signals as well as fluorescence emissions in the near infrared regions where most biomolecules are transparent. Such signals do not blink or photobleach under prolonged excitation. which is advantageous to optical nano-bio marker applications. In this paper, single walled carbon nanotubes are conjugated with specific types of single-stranded DNA in order to detect oligonucleotides of corresponding complimentary sequences. Dot blotting experiments and comparative Raman spectroscopy observations demonstrated excellent sensitivity and specificity of carbon nanotube-DNA probes. The results show the possibility of using SWNT as generic nano-bio markers for the precise detection of specific kinds of genes.

Design and Synthesis of Novel Rhodamine-based Chemosensor Probe Toward Cu2+ Cation

  • Son, Young-A
    • 한국염색가공학회지
    • /
    • 제26권1호
    • /
    • pp.7-12
    • /
    • 2014
  • Nowdays, fluorescent rhodamine chemosensors have attracted a worldwide interest due to its ability to selectively detect heavy and transition metal cations. Due to the importance in environmental and biological toxic effects, the developments of fluorescent chemosensors have been received considerable attention in recent. Especially, a rhodamine-based chemosensor probes have been proved to be useful by exhibiting the efficient "off-on" fluorescence switching toward selected metal cations. This fluorophore can undergo the transformation from non-fluorescent and colorless spirolactam derivative to fluorescent ring-open form. In this study, a new fluorescent chemosensor was synthesized using rhodamine B through two-step procedures, and its selectivity and related optical property were characterized. Selectivity and sensitivity was found toward $Cu^{2+}$ guest molecules and then related optical properties of rhodamine B based fluorescent chemosensor compound were characterized using discussed. In addition, computational calculation was used to determine the HOMO/LUMO values.

High-throughput SNP Genotyping by Melting Curve Analysis for Resistance to Southern Root-knot Nematode and Frogeye Leaf Spot in Soybean

  • Ha, Bo-Keun;Boerma, H. Roger
    • Journal of Crop Science and Biotechnology
    • /
    • 제11권2호
    • /
    • pp.91-100
    • /
    • 2008
  • Melting curve analysis of fluorescently labeled DNA fragments is used extensively for genotyping single nucleotide polymorphism(SNP). Here, we evaluated a SNP genotyping method by melting curve analysis with the two probe chemistries in a 384-well plate format on a Roche LightCycler 480. The HybProbe chemistry is based on the fluorescence resonance energy transfer(FRET) and the SimpleProbe chemistry uses a terminal self-quenching fluorophore. We evaluated FRET HybProbes and SimpleProbes for two SNP sites closely linked to two quantitative trait loci(QTL) for southern root-knot nematode resistance. These probes were used to genotype the two parents and 94 $F_2$ plants from the cross of PI 96354$\times$Bossier. The SNP genotypes of all samples determined by the LightCycler software agreed with previously determined SSR genotypes and the SNP genotypes determined on a Luminex 100 flow cytometry instrument. Multiplexed HybProbes for the two SNPs showed a 98.4% success rate and 100% concordance between repeats two of the same 96 DNA samples. Also, we developed a HybProbe assay for the Rcs3 gene conditioning broad resistance to the frogeye leaf spot(FLS) disease. The LightCycler 480 provides rapid PCR on 384-well plate and allows simultaneous amplification and analysis in approximately 2 hours without any additional steps after amplification. This allowed for a reduction of the potential contamination of PCR products, simplicity, and enablement of a streamlined workflow. The melting curve analysis on the LightCycler 480 provided high-throughput and rapid SNP genotyping and appears highly effective for marker-assisted selection in soybean.

  • PDF

Generation of FISH Probes Using Laser Microbeam Microdissection and Application to Clinical Molecular Cytogenetics

  • Shim, Sung-Han;Kyhm, Jee-Hong;Chung, Sung-Ro;Kim, Seung-Ryong;Park, Moon-Il;Lee, Chul-Hoon;Cho, Youl-Hee
    • Journal of Microbiology and Biotechnology
    • /
    • 제17권7호
    • /
    • pp.1079-1082
    • /
    • 2007
  • Chromosome microdissection and the reverse FISH technique is one of the most useful methods for the identification of structurally abnormal chromosomes. In particular, the laser microbeam microdissection (LMM) method allows rapid isolation of a target chromosome or a specific region of chromosomes without damage of genetic materials and contamination. Isolated chromosomes were directly amplified by the degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), and then the FISH probes labeled with spectrum green- or spectrum red-dUTP were generated by nick-translation. Whole chromosome painting (WCP) probes were successfully generated from only 5 copies of the chromosome. With this method, we produced 24 WCP probes for each human chromosome. We also tried to characterize a marker chromosome, which seemed to be originated from chromosome 11 on conventional banding technique. The marker chromosomes were isolated by the LMM method and analyzed by reverse FISH. We elucidated that the marker chromosome was originated from the short arm of chromosome 5 ($5p11{\to}pter$). A fully automated and computer-controlled LMM method is a very simple laboratory procedure, and enables rapid and precise characterization of various chromosome abnormalities.

Age Prediction in the Chickens Using Telomere Quantity by Quantitative Fluorescence In situ Hybridization Technique

  • Kim, Y.J.;Subramani, V.K.;Sohn, S.H.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • 제24권5호
    • /
    • pp.603-609
    • /
    • 2011
  • Telomeres are special structures at the ends of eukaryotic chromosomes. Vertebrate telomeres consist of tandem repeats of conserved TTAGGG sequence and associated proteins. Birds are interesting models for molecular studies on aging and cellular senescence because of their slow aging rates and longer life spans for their body size. In this longitudinal study, we explored the possibility of using telomeres as an age-marker to predict age in Single Comb White Leghorn layer chickens. We quantified the relative amount of telomeric DNA in isolated peripheral blood lymphocytes by the Quantitative Fluorescence in situ Hybridization technique on interphase nuclei (IQ FISH) using telomere-specific DNA probes. We found that the amount of telomeric DNA (ATD) reduced significantly with an increase in chronological age of the chicken. Especially, the telomere shortening rates are greatly increased in growing individuals compared to laying and old-aged individuals. Therefore, using the ATD values obtained by IQ FISH we established the possibility of age prediction in chickens based on the telomere theory of aging. By regression analysis of the ATD values at each age interval, we formulated an equation to predict the age of chickens. In conclusion, the telomeric DNA values by IQ FISH analyses can be used as an effective age-marker in predicting the chronological age of chickens. The study has implications in the breeding and population genetics of poultry, especially the reproductive potential.

DNA Dynamics: a Fluorescence Resonance Energy Transfer Study Using a Long-Lifetime Metal-Ligand Complex

  • Kang, Jung-Sook;Lakowicz, Joseph-R.;Piszczek, Grzegorz
    • Archives of Pharmacal Research
    • /
    • 제25권2호
    • /
    • pp.143-150
    • /
    • 2002
  • Fluorescent probes bound to DNA typically display nanosecond decay times and reveal only nanosecond motions. We extend the time range of measurable DNA dynamics using $[Ru(pby)_2(dppz)]^{2+}$ (bpy=2.2'-bipyridine, dppz=dipyrido[3,2-a2',3'-c]phenazine) (RuBD) which displays a mean lifetime near 90 ns. To test the usefulness of RuBD as a probe for diffusive processes in calf thymus DNA, we compared the efficiencies of fluorescence resonance energy transfer (FRET) using three donors which display lifetimes near 5 ns for acridine orange (AO), 22 ns for ethidum bromide (EB) and 92 ns for RuBD, with nile blue (NB) as the acceptor. The F rster distances for AO-NB, EB-NB and RuBD-NB donor-acceptor pairs were 42.3, 52.3, and $30.6{\;}{\AA}$, respectively. All three donors showed dramatic decreases in fluorescence intensities and more rapid intensity decays with increasing NB concentrations. The intensity decays of AO and EB in the presence of varying concentrations of NB were satisfactorily described by the one-dimensional FRET model without diffusion (Blumen and Manz, 1979). In the case of the long-lifetime donor RuBD, the experimental phase and modulation somewhat deviated from the recovered values computed from this model. The recovered NB concentrations and FRET efficiencies from the model were slightly larger than the expected values, however, the recovered and expected values did not show a significant difference. Thus, it is suggested that the lifetime of RuBD is too short to measure diffusive processes in calf thymus DNA.

양자점 기반 다중 바이오마커 검출법의 연구동향 (Recent Progress in Multiplexed Detection of Biomarkers Based on Quantum Dots)

  • 김예린;최유림;김봉근;나현빈
    • 공업화학
    • /
    • 제33권5호
    • /
    • pp.451-458
    • /
    • 2022
  • 반도체 양자점은 우수한 형광 특성을 가진 광학 탐침자로 생명-의학 영상화 기술 및 바이오센싱 분야에서 광범위하게 활용되고 있다. 양자점은 넓은 광흡수 에너지띠, 좁은 형광 에너지띠와 같은 광학 특성을 가지므로 서로 다른 형광 파장을 지닌 양자점을 조합해 다종의 신호를 생성할 수 있도록 구성하면 복수의 바이오마커를 동시에 검출할 수 있다. 본 총설에서는 이와 같은 다중 검출 분석법에서의 양자점 및 이에 기반한 양자점 나노비드가 가지는 장점과 활용 사례를 기술하고 다중 형광 바이오마커 검출법의 최근 개발 동향 및 개선사항을 요약 정리하였다. 특히 양자점을 활용한 형광-결합 면역흡착 분석법, 양자점 나노비드를 이용한 면역크로마토그래피 분석법 등 면역 분석법에서의 신호 전환 소재 디자인을 중심으로 최근의 연구 결과를 검토하였다. 정확성과 민감도가 우수한 다중 바이오마커 검출 기술이 확보된 데이터를 처리하고 해석하는 인공지능 알고리즘과 결합될 경우 질병의 조기 진단을 포함한 다양한 분야에 활용가능한 새로운 검출 플랫폼의 개발로 이어질 것으로 기대된다.