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Sample records for fluorescence intensity multiple

  1. Conversion of logarithmic channel numbers into relative linear fluorescence intensity.

    Science.gov (United States)

    Schmid, I; Schmid, P; Giorgi, J V

    1988-11-01

    We describe a simple, reproducible, and generally applicable method to assess the performance of log amplifiers by using a fluorescent sample that provides multiple peaks of different intensities. The channel differences between multiple peaks are used to evaluate the logarithmic behavior of the fluorescence signal amplifier on the flow cytometer. A calibration curve can be created to correct the channel numbers for deviations from true logarithmic behavior and then convert data into relative linear intensities. By using these linear fluorescent intensities, we compared the capacity of different antisera against HIV-1 (human immunodeficiency virus type 1) peptides to inhibit the binding of HIV-1 to CEM, a CD4-positive T-cell line. A wide range of applications for this calibration procedure can be envisioned and the method is valuable for monitoring instrument performance over time.

  2. Intense fluorescence of Au 20

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Chongqi; Harbich, Wolfgang; Sementa, Luca; Ghiringhelli, Luca; Aprá, Edoardo; Stener, Mauro; Fortunelli, Alessandro; Brune, Harald

    2017-08-21

    Ligand-protected Au clusters are non-bleaching fluorescence markers in bio- and medical applications. We show that their fluorescence is an intrinsic property of the Au cluster itself. We find a very intense and sharp fluorescence peak located at λ =739.2 nm (1.68 eV) for Au20 clusters in a Ne matrix held at 6 K. The fluorescence reflects the HOMO-LUMO diabatic bandgap of the cluster. The cluster shows a very rich absorption fine structure reminiscent of well defined molecule-like quantum levels. These levels are resolved since Au20 has only one stable isomer (tetrahedral), therefore our sample is mono-disperse in cluster size and conformation. Density-functional theory (DFT) and time-dependent DFT calculations clarify the nature of optical absorptionand predict both main absorption peaks and intrinsic fluorescence in good agreement with experiment.

  3. High Intensity Exercise in Multiple Sclerosis

    DEFF Research Database (Denmark)

    Wens, Inez; Dalgas, Ulrik; Vandenabeele, Frank

    2015-01-01

    Introduction Low-to-moderate intensity exercise improves muscle contractile properties and endurance capacity in multiple sclerosis (MS). The impact of high intensity exercise remains unknown. Methods Thirty-four MS patients were randomized into a sedentary control group (SED, n = 11) and 2...

  4. Waveguide evanescent field fluorescence microscopy: Thin film fluorescence intensities and its application in cell biology

    Science.gov (United States)

    Hassanzadeh, Abdollah; Nitsche, Michael; Mittler, Silvia; Armstrong, Souzan; Dixon, Jeff; Langbein, Uwe

    2008-06-01

    We demonstrate an inexpensive alternative to total internal reflection fluorescence microscopy. A method for imaging ultrathin films and living cells located on waveguides—illuminated with their evanescent fields—is introduced. An extensive analysis of ion-exchanged waveguides focusing on their application as microscopy substrates for studying interfacial phenomena is presented. Experimental results are in excellent agreement with the simulations. As an application osteoblasts (bone matrix forming cells) and ultrathin Langmuir-Blodgett films were imaged. The fluorescence intensity has been used to determine the cell attachment.

  5. Strongly intensive measures for multiplicity fluctuations

    Science.gov (United States)

    Begun, V. V.; Konchakovski, V. P.; Gorenstein, M. I.; Bratkovskaya, E. L.

    2013-04-01

    The two recently proposed families of strongly intensive measures of fluctuations and correlations are studied within the hadron-string-dynamics (HSD) transport approach to nucleus-nucleus collisions. We consider the measures ΔKπ and ΣKπ for kaon and pion multiplicities in Au+Au collisions in a wide range of collision energies and centralities. These strongly intensive measures appear to cancel the participant number fluctuations. This allows to enlarge the centrality window in the analysis of event-by-event fluctuations for up to at least 10% of the most central collisions. We also present a comparison of the HSD results with the data of the NA49 and STAR Collaborations. HSD describes ΣKπ reasonably well. However, the HSD results depend monotonously on collision energy and do not reproduce the bump-dip structure of ΔKπ observed from the NA49 data in the region of the center of mass energy of the nucleon pair \\sqrt{s_{NN}}= 8{--}12 GeV. This observation deserves further study. The origin of this ‘structure’ is not connected with simple geometrical or limited acceptance effects, as these effects are taken into account in HSD simulations.

  6. High-intensity light-emitting diode vs fluorescent tubes for intensive phototherapy in neonates.

    Science.gov (United States)

    Sherbiny, Hanan S; Youssef, Doaa M; Sherbini, Ahmad S; El-Behedy, Rabab; Sherief, Laila M

    2016-05-01

    Special blue fluorescent tubes are recommended by the American Academy of Pediatrics (AAP) as the most effective light source for lowering serum bilirubin. A high-intensity light-emitting diode ('super LED') could render intensive phototherapy more effective than the above conventional methods. This study compared the efficacy and safety of a high-intensity light-emitting diode bed vs conventional intensive phototherapy with triple fluorescent tube units as a rescue treatment for severe unconjugated neonatal hyperbilirubinaemia. This was a randomised, prospective trial. Two hundred jaundiced neonates ≥ 35 weeks gestation who met the criteria for intensive phototherapy as per AAP guidelines were randomly assigned to be treated either with triple fluorescent tube units (group 1, n = 100) or a super LED bed (group 2, n = 100). The outcome was the avoidance of exchange transfusion by successful control of hyperbilirubinaemia. Statistically significant higher success rates of intensive phototherapy were achieved among neonates treated with super LED (group 2) than in those treated conventionally (group 1) (87% vs 64%, P = 0.003). Significantly higher 'bilirubin decline' rates were reported in both haemolytic and non-haemolytic subgroups treated with the super LED bed compared with a similar sub-population in the conventionally treated group. Comparable numbers of neonates in both groups developed rebound jaundice (8% vs 10% of groups 1 and 2, respectively). Side-effects were mild in both groups, but higher rates of hyperthermia (12% vs 0%, P = 0.03), dehydration (8% vs 2%, P = 0.26) and skin rash (39% vs 1%, P = 0.002) were reported in the fluorescent tubes-treated group compared with the LED group. Super LED is a safe rescue treatment for severe neonatal hyperbilirubinaemia, and its implementation may reduce the need for exchange transfusion.

  7. Multiple stimulated emission fluorescence photoacoustic sensing and spectroscopy

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    Li, Gaoming [School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore 639798 (Singapore); Key Laboratory of OptoElectronic Science and Technology for Medicine, Ministry of Education, Fujian Normal University, Fuzhou 350007 (China); Gao, Fei; Feng, Xiaohua; Zheng, Yuanjin, E-mail: yjzheng@ntu.edu.sg [School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore 639798 (Singapore); Qiu, Yishen [Key Laboratory of OptoElectronic Science and Technology for Medicine, Ministry of Education, Fujian Normal University, Fuzhou 350007 (China)

    2016-07-04

    Multiple stimulated emission fluorescence photoacoustic (MSEF-PA) phenomenon is demonstrated in this letter. Under simultaneous illumination of pumping light and stimulated emission light, the fluorescence emission process is speeded up by the stimulated emission effect. This leads to nonlinear enhancement of photoacoustic signal while the quantity of absorbed photons is more than that of fluorescent molecules illuminated by pumping light. The electronic states' specificity of fluorescent molecular can also be labelled by the MSEF-PA signals, which can potentially be used to obtain fluorescence excitation spectrum in deep scattering tissue with nonlinearly enhanced photoacoustic detection. In this preliminary study, the fluorescence excitation spectrum is reconstructed by MSEF-PA signals through sweeping the wavelength of exciting light, which confirms the theoretical derivation well.

  8. Quantitative Brightness Analysis of Fluorescence Intensity Fluctuations in E. Coli.

    Science.gov (United States)

    Hur, Kwang-Ho; Mueller, Joachim D

    2015-01-01

    The brightness measured by fluorescence fluctuation spectroscopy specifies the average stoichiometry of a labeled protein in a sample. Here we extended brightness analysis, which has been mainly applied in eukaryotic cells, to prokaryotic cells with E. coli serving as a model system. The small size of the E. coli cell introduces unique challenges for applying brightness analysis that are addressed in this work. Photobleaching leads to a depletion of fluorophores and a reduction of the brightness of protein complexes. In addition, the E. coli cell and the point spread function of the instrument only partially overlap, which influences intensity fluctuations. To address these challenges we developed MSQ analysis, which is based on the mean Q-value of segmented photon count data, and combined it with the analysis of axial scans through the E. coli cell. The MSQ method recovers brightness, concentration, and diffusion time of soluble proteins in E. coli. We applied MSQ to measure the brightness of EGFP in E. coli and compared it to solution measurements. We further used MSQ analysis to determine the oligomeric state of nuclear transport factor 2 labeled with EGFP expressed in E. coli cells. The results obtained demonstrate the feasibility of quantifying the stoichiometry of proteins by brightness analysis in a prokaryotic cell.

  9. Quantitative Brightness Analysis of Fluorescence Intensity Fluctuations in E. Coli.

    Directory of Open Access Journals (Sweden)

    Kwang-Ho Hur

    Full Text Available The brightness measured by fluorescence fluctuation spectroscopy specifies the average stoichiometry of a labeled protein in a sample. Here we extended brightness analysis, which has been mainly applied in eukaryotic cells, to prokaryotic cells with E. coli serving as a model system. The small size of the E. coli cell introduces unique challenges for applying brightness analysis that are addressed in this work. Photobleaching leads to a depletion of fluorophores and a reduction of the brightness of protein complexes. In addition, the E. coli cell and the point spread function of the instrument only partially overlap, which influences intensity fluctuations. To address these challenges we developed MSQ analysis, which is based on the mean Q-value of segmented photon count data, and combined it with the analysis of axial scans through the E. coli cell. The MSQ method recovers brightness, concentration, and diffusion time of soluble proteins in E. coli. We applied MSQ to measure the brightness of EGFP in E. coli and compared it to solution measurements. We further used MSQ analysis to determine the oligomeric state of nuclear transport factor 2 labeled with EGFP expressed in E. coli cells. The results obtained demonstrate the feasibility of quantifying the stoichiometry of proteins by brightness analysis in a prokaryotic cell.

  10. A portable fluorescent sensing system using multiple LEDs

    Science.gov (United States)

    Shin, Young-Ho; Barnett, Jonathan Z.; Gutierrez-Wing, M. Teresa; Rusch, Kelly A.; Choi, Jin-Woo

    2017-02-01

    This paper presents a portable fluorescent sensing system that utilizes different light emitting diode (LED) excitation lights for multiple target detection. In order to identify different analytes, three different wavelengths (385 nm, 448 nm, and 590 nm) of excitation light emitting diodes were used to selectively stimulate the target analytes. A highly sensitive silicon photomultiplier (SiPM) was used to detect corresponding fluorescent signals from each analyte. Based on the unique fluorescent response of each analyte, it is possible to simultaneously differentiate one analyte from the other in a mixture of target analytes. A portable system was designed and fabricated consisting of a display module, battery, data storage card, and sample loading tray into a compact 3D-printed jig. The portable sensor system was demonstrated for quantification and differentiation of microalgae (Chlorella vulgaris) and cyanobacteria (Spirulina) by measuring fluorescent responses of chlorophyll a in microalgae and phycocyanin in cyanobacteria. Obtained results suggest that the developed portable sensor system could be used as a generic fluorescence sensor platform for on-site detection of multiple analytes of interest.

  11. Fluorescence intensity studies of Triassic acritarchs from the Yanchang Formation in Ordos basin,northwestern China

    Institute of Scientific and Technical Information of China (English)

    JI Liming; MENG Fanwei; XU Jinli

    2007-01-01

    Fluorescence properties of Early Cambrian acritarchs were investigated using Leica das Mikroskop (DM)microscopy with a mercury lamp.Well-preserved autoflurescence properties show a correlation between acritarchs morphology and the intensity of emitted fluorescence.In accordance with the fluorescence intensity of organic cell walls,two groups ofmicrofossils were distinguished.Results of observation in this study,which are consistent with those of the previous foreign studies,are in good agreement with regular difference in autofluorescence intensity among palynomorphs reported by McPhilemy (1998).Spores and algae,including Botryococcus,have very bright fluorescence while acritarchs often show less intense fluorescence.Dark brown microfossils have been reworked,and have little or no fluorescence.

  12. Dependence of fluorescence intensity on the spectral overlap between fluorophores and plasmon resonant single silver nanoparticles.

    Science.gov (United States)

    Chen, Yeechi; Munechika, Keiko; Ginger, David S

    2007-03-01

    We investigate the fluorescence from dyes coupled to individual DNA-functionalized metal nanoparticles. We use single-particle darkfield scattering and fluorescence microscopy to correlate the fluorescence intensity of the dyes with the localized surface plasmon resonance (LSPR) spectra of the individual metal nanoparticles to which they are attached. For each of three different dyes, we observe a strong correlation between the fluorescence intensity of the dye and the degree of spectral overlap with the plasmon resonance of the nanoparticle. On average, we observe the brightest fluorescence from dyes attached to metal nanoparticles that have a LSPR scattering peak approximately 40-120 meV higher in energy than the emission peak of the fluorophore. These results should prove useful for understanding and optimizing metal-enhanced fluorescence.

  13. Coordinate-targeted fluorescence nanoscopy with multiple off states

    Science.gov (United States)

    Danzl, Johann G.; Sidenstein, Sven C.; Gregor, Carola; Urban, Nicolai T.; Ilgen, Peter; Jakobs, Stefan; Hell, Stefan W.

    2016-02-01

    Far-field super-resolution fluorescence microscopy discerns fluorophores residing closer than the diffraction barrier by briefly transferring them in different (typically ON and OFF) states before detection. In coordinate-targeted super-resolution variants, such as stimulated emission depletion (STED) microscopy, this state difference is created by the intensity minima and maxima of an optical pattern, causing all fluorophores to assume the off state, for instance, except at the minima. Although strong spatial confinement of the on state enables high resolution, it also subjects the fluorophores to excess intensities and state cycles at the maxima. Here, we address these issues by driving the fluorophores into a second off state that is inert to the excess light. By using reversibly switchable fluorescent proteins as labels, our approach reduces bleaching and enhances resolution and contrast in live-cell STED microscopy. Using two or more transitions to off states is a useful strategy for augmenting the power of coordinate-targeted super-resolution microscopy.

  14. Analysis of root surface properties by fluorescence/Raman intensity ratio.

    Science.gov (United States)

    Nakamura, Shino; Ando, Masahiro; Hamaguchi, Hiro-O; Yamamoto, Matsuo

    2017-07-25

    The aim of this study is to evaluate the existence of residual calculus on root surfaces by determining the fluorescence/Raman intensity ratio. Thirty-two extracted human teeth, partially covered with calculus on the root surface, were evaluated by using a portable Raman spectrophotometer, and a 785-nm, 100-mW laser was applied for fluorescence/Raman excitation. The collected spectra were normalized to the hydroxyapatite Raman band intensity at 960 cm(-1). Raman spectra were recorded from the same point after changing the focal distance of the laser and the target radiating angle. In seven teeth, the condition of calculus, cementum, and dentin were evaluated. In 25 teeth, we determined the fluorescence/Raman intensity ratio following three strokes of debridement. Raman spectra collected from the dentin, cementum, and calculus were different. After normalization, spectra values were constant. The fluorescence/Raman intensity ratio of calculus region showed significant differences compared to the cementum and dentin (p < 0.05). The fluorescence/Raman intensity ratio decreased with calculus debridement. For this analysis, the delta value was defined as the difference between the values before and after three strokes, with the final 2 delta values close to zero, indicating a gradual asymptotic curve and the change in intensity ratio approximating that of individual constants. Fluorescence/Raman intensity ratio was effectively used to cancel the angle- and distance-dependent fluctuations of fluorescence collection efficiency during measurement. Changes in the fluorescence/Raman intensity ratio near zero suggested that cementum or dentin was exposed, and calculus removed.

  15. A precise Boltzmann distribution law for the fluorescence intensity ratio of two thermally coupled levels

    Science.gov (United States)

    Qin, Feng; Zhao, Hua; Cai, Wei; Zhang, Zhiguo; Cao, Wenwu

    2016-06-01

    Noncontact monitoring temperature is very important in modern medicine, science, and technologies. The fluorescence intensity ratio (FIR) technique based on the Boltzmann distribution law exhibits excellent application potential, but the observed FIR deviates from the Boltzmann distribution law in the low temperature range. We propose a fluorescence intensity ratio relation FIR* = ηFIR by introducing a quantity η representing thermal population degree, which can be obtained from measured fluorescence decay curves of the upper emitting level. Using Eu3+ as an example, the method is confirmed that the deviated FIR is able to be corrected and return to follow the Boltzmann law.

  16. Development of a software for reconstruction of X-ray fluorescence intensity maps

    Energy Technology Data Exchange (ETDEWEB)

    Almeida, Andre Pereira de; Braz, Delson; Mota, Carla Lemos, E-mail: apalmeid@gmail.co, E-mail: delson@lin.ufrj.b, E-mail: clemos@con.ufrj.b [Coordenacao dos Programas de Pos-graduacao de Engenharia (COPPE/UFRJ), Rio de Janeiro, RJ (Brazil). Programa de Engenharia Nuclear; Oliveira, Luis Fernando de; Barroso, Regina Cely; Pinto, Nivia Graciele Villela, E-mail: cely@uerj.b, E-mail: lfolive@uerj.b, E-mail: nitatag@gmail.co [Universidade do Estado do Rio de Janeiro (UERJ), Rio de Janeiro, RJ (Brazil). Inst. de Fisica; Cardoso, Simone Coutinho, E-mail: simone@if.ufrj.b [Universidade Federal do Rio de Janeiro (UFRJ), RJ (Brazil). Inst. de Fisica; Moreira, Silvana, E-mail: silvana@fec.unicamp.b [Universidade Estadual de Campinas (UNICAMP), SP (Brazil). Faculdade de Engenharia Civil, Arquitetura e Urbanismo

    2009-07-01

    The technique of X-ray fluorescence (XRF) using SR microbeams is a powerful analysis tool for studying elemental composition in several samples. One application of this technique is the analysis done through the mapping of chemical elements forming a matrix of data. The aim of this work is the presentation of the program MapXRF, an in-house software designed to optimize the processing and mapping of fluorescence intensities data. This program uses spectra generated by QXAS as input data and separates the intensities of each chemical element found in the fluorescence spectra in files themselves. From these files, the program generates the intensity maps that can be visualized in any program of treatment of images. The proposed software was tested using fluorescence data obtained in the XRF beamline of XRF at Synchrotron Light National Laboratory (LNLS), Brazil. Automatic 2D scans were performed and element distribution maps were obtained in the form of a matrix of data. (author)

  17. Influence of the excitation light intensity on the rate of fluorescence quenching reactions: pulsed experiments.

    Science.gov (United States)

    Angulo, Gonzalo; Milkiewicz, Jadwiga; Kattnig, Daniel; Nejbauer, Michał; Stepanenko, Yuriy; Szczepanek, Jan; Radzewicz, Czesław; Wnuk, Paweł; Grampp, Günter

    2017-02-22

    The effect of multiple light excitation events on bimolecular photo-induced electron transfer reactions in liquid solution is studied experimentally. It is found that the decay of fluorescence can be up to 25% faster if a second photon is absorbed after a first cycle of quenching and recombination. A theoretical model is presented which ascribes this effect to the enrichment of the concentration of quenchers in the immediate vicinity of fluorophores that have been previously excited. Despite its simplicity, the model delivers a qualitative agreement with the observed experimental trends. The original theory by Burshtein and Igoshin (J. Chem. Phys., 2000, 112, 10930-10940) was created for continuous light excitation though. A qualitative extrapolation from the here presented pulse experiments to the continuous excitation conditions lead us to conclude that in the latter the order of magnitude of the increase of the quenching efficiency upon increasing the light intensity of excitation, must also be on the order of tens of percent. These results mean that the rate constant for photo-induced bimolecular reactions depends not only on the usual known factors, such as temperature, viscosity and other properties of the medium, but also on the intensity of the excitation light.

  18. Fluorescence intensity and bright spot analyses using a confocal microscope for photodynamic diagnosis of brain tumors.

    Science.gov (United States)

    Yoneyama, Takeshi; Watanabe, Tetsuyo; Kagawa, Hiroyuki; Hayashi, Yutaka; Nakada, Mitsutoshi

    2017-03-01

    In photodynamic diagnosis using 5-aminolevulinic acid (5-ALA), discrimination between the tumor and normal tissue is very important for a precise resection. However, it is difficult to distinguish between infiltrating tumor and normal regions in the boundary area. In this study, fluorescent intensity and bright spot analyses using a confocal microscope is proposed for the precise discrimination between infiltrating tumor and normal regions. From the 5-ALA-resected brain tumor tissue, the red fluorescent and marginal regions were sliced for observation under a confocal microscope. Hematoxylin and eosin (H&E) staining were performed on serial slices of the same tissue. According to the pathological inspection of the H&E slides, the tumor and infiltrating and normal regions on confocal microscopy images were investigated. From the fluorescent intensity of the image pixels, a histogram of pixel number with the same fluorescent intensity was obtained. The fluorescent bright spot sizes and total number were compared between the marginal and normal regions. The fluorescence intensity distribution and average intensity in the tumor were different from those in the normal region. The probability of a difference from the dark enhanced the difference between the tumor and the normal region. The bright spot size and number in the infiltrating tumor were different from those in the normal region. Fluorescence intensity analysis is useful to distinguish a tumor region, and a bright spot analysis is useful to distinguish between infiltrating tumor and normal regions. These methods will be important for the precise resection or photodynamic therapy of brain tumors. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Fluorescence enhancement and multiple protein detection in ZnO nanostructure microfluidic devices.

    Science.gov (United States)

    Sang, Chen-Hsiang; Chou, Shu-Jen; Pan, F M; Sheu, Jeng-Tzong

    2016-01-15

    In this study, different morphological ZnO nanostructures, those of sharp nanowires (NWs), rod NWs, and hexahedral-puncheon nanostructures, were grown in microfluidic channels on the same glass substrate. Characterizations of correspondent biomolecule binding properties were simulated and demonstrated. The surface was modified using 3-ammineopropyl-triethoxysilane (3-APTES) and biotin-N-hydroxysuccinimide ester (NHS-biotin). Different concentrations (4.17pM to 41.7nM) of dye-conjugated streptavidin were simultaneously infused through the second microfluidic channels, which lie 90° from the first microfluidic channels. The florescent intensity at the crossover areas showed good agreement with simulations, with sharp ZnO NWs exhibiting the largest dynamic range and the highest fluorescent intensity. We further characterize correspondent protein detection using sharp ZnO NWs. The surfaces of these ZnO NWs were modified with mouse immunoglobulin G (IgG), infused through the second microfluidic channels with dye-conjugated (Alexa 546) anti-mouse IgG in different concentrations. Concentrations ranging from 417fM to 41.7nM can be resolved using sharp ZnO NWs. Finally, multiple protein detection was demonstrated using a five-by-eight microfluidic channel array. Fluorescence images present clear multiple detections at the crossover areas when using the sharp ZnO NWs for simultaneous dye-conjugated anti-mouse IgG and dye-conjugated anti-rabbit IgG (Alexa 647) detection.

  20. Multiple signal classification algorithm for super-resolution fluorescence microscopy

    Science.gov (United States)

    Agarwal, Krishna; Macháň, Radek

    2016-12-01

    Single-molecule localization techniques are restricted by long acquisition and computational times, or the need of special fluorophores or biologically toxic photochemical environments. Here we propose a statistical super-resolution technique of wide-field fluorescence microscopy we call the multiple signal classification algorithm which has several advantages. It provides resolution down to at least 50 nm, requires fewer frames and lower excitation power and works even at high fluorophore concentrations. Further, it works with any fluorophore that exhibits blinking on the timescale of the recording. The multiple signal classification algorithm shows comparable or better performance in comparison with single-molecule localization techniques and four contemporary statistical super-resolution methods for experiments of in vitro actin filaments and other independently acquired experimental data sets. We also demonstrate super-resolution at timescales of 245 ms (using 49 frames acquired at 200 frames per second) in samples of live-cell microtubules and live-cell actin filaments imaged without imaging buffers.

  1. Improving accuracy and capabilities of X-ray fluorescence method using intensity ratios

    Science.gov (United States)

    Garmay, Andrey V.; Oskolok, Kirill V.

    2017-04-01

    An X-ray fluorescence analysis algorithm is proposed which is based on a use of ratios of X-ray fluorescence lines intensities. Such an analytical signal is more stable and leads to improved accuracy. Novel calibration equations are proposed which are suitable for analysis in a broad range of matrix compositions. To apply the algorithm to analysis of samples containing significant amount of undetectable elements a use of a dependence of a Rayleigh-to-Compton intensity ratio on a total content of these elements is suggested. The technique's validity is shown by analysis of standard steel samples, model metal oxides mixture and iron ore samples.

  2. Improving accuracy and capabilities of X-ray fluorescence method using intensity ratios

    Energy Technology Data Exchange (ETDEWEB)

    Garmay, Andrey V., E-mail: andrew-garmay@yandex.ru; Oskolok, Kirill V.

    2017-04-15

    An X-ray fluorescence analysis algorithm is proposed which is based on a use of ratios of X-ray fluorescence lines intensities. Such an analytical signal is more stable and leads to improved accuracy. Novel calibration equations are proposed which are suitable for analysis in a broad range of matrix compositions. To apply the algorithm to analysis of samples containing significant amount of undetectable elements a use of a dependence of a Rayleigh-to-Compton intensity ratio on a total content of these elements is suggested. The technique's validity is shown by analysis of standard steel samples, model metal oxides mixture and iron ore samples.

  3. Comparative study of the fluorescence intensity of dental composites and human teeth submitted to artificial aging.

    Science.gov (United States)

    Jablonski, Tatiana; Takahashi, Marcos Kenzo; Brum, Rafeal Torres; Rached, Rodrigo Nunes; Souza, Evelise M

    2014-01-01

    The aim of this study was to evaluate quantitatively the fluorescence of resin composites and human teeth, and to determine the stability of fluorescence after aging. Ten specimens were built using a 1 mm thick increment of dentin composite overlapped by a 0.5 mm thick increment of enamel composite. Ten sound human molars were sectioned and silicon carbide-polished to obtain enamel and dentin slabs 1.5 mm in thickness. Fluorescence measurements were carried out by a fluorescence spectrophotometer before and after thermocycling (2000 cycles, 5°C and 55°C). One-way analysis of variance (ANOVA) with repeated measures and Tukey's test were performed at a significance level of 5%. Most of the tested composites showed significant differences in fluorescence both before and after aging (P composite whose fluorescence was similar to that of human teeth at both periods of evaluation (P > 0.05), and was the only composite that showed comparable results of fluorescence to the tooth structure before and after thermocycling. With the exception of Filtek Supreme, there were significant reductions in fluorescence intensity for all the tested composites (P < 0.05).

  4. Extracting molecular Hamiltonian structure from time-dependent fluorescence intensity data

    OpenAIRE

    Brif, Constantin; Rabitz, Herschel

    2000-01-01

    We propose a formalism for extracting molecular Hamiltonian structure from inversion of time-dependent fluorescence intensity data. The proposed method requires a minimum of \\emph{a priori} knowledge about the system and allows for extracting a complete set of information about the Hamiltonian for a pair of molecular electronic surfaces.

  5. Major Metabolite Levels of Preoperative Proton Magnetic Resonance Sectroscopy and Intraoperative Fluorescence Intensity in Glioblastoma.

    Science.gov (United States)

    Tian, Hai-Long; Zu, Yu-Liang; Wang, Chao-Chao; Lin, Tao; Guo, Zhen-Tao; Jiang, Bin; Yin, Xin; Guo, Wen-Qiang; Wang, Zhi-Gang

    2017-08-20

    Objective To compare the intraoperative major metabolite level of preoperative proton magnetic resonance spectroscopy((1)H-MRS)and fluorescence intensity marked with fluorescein sodium(FLs)in glioblastoma(GBM)and thus provide an objective basis for fluorescence surgical treatment of GBM. Methods All newly diagnosed patients by plain and enhanced magnetic resonance imaging from the April 1,2014 to December 31,2015 were enrolled in this study.All of them received (1)H-MRS and marked with FLs.The expression of Ki67 in tumor boundary were confirmed by postoperative pathology and determined by immunostaining assay.The relationship between (1)H-MRS metabolite levels and tumor fluorescence intensity was analyzed. Results Totally 33 patients were included in the study.Preoperative (1)H-MRS revealed high-grade gliomas in 25 cases.The N-acetylaspartate(NAA)decreased significantly and choline(Cho)increased significantly in high-grade gliomas.The ratios of Cho/NAA,NAA/creatine(Cr),and Cho/Cr significantly differed in different tumor regions(P=0.02,P=0.01,and P=0.00,respectively).Surgical results were marked with FLs intraoperatively.Tissue fluorescence were clearly seen.There were 29 patients undergoing total resection and 4 cases undergoing subtotal resection.No acute encephalocele occured after operation,while 2 patients suffered from epilepsy.Postoperative pathology results included:28 cases were diagnosed as GBM(22 cases consistent with (1)H-MRS diagnosis).The results of GBM fluorescence imaging included:the level of fluorescence intensity in tumor parenchyma was significantly higher than that in tumor boundary and peritumoral edema(P=0.01).The result of (1)H-MRS metabolite analysis included:The kurtosis of NAA and of Cho and the ratio of Cho/NAA were significantly different according the fluorescence intensity in tumor parenchyma(P=0.01,P=0.02,and P=0.01).While there was no difference in the kurtosis of NAA,the kurtosis of Cho and the ratio of Cho/NAA were significantly

  6. Effect of excitation intensity on fluorescence spectra in ZnO nanostructures and its origin

    Institute of Scientific and Technical Information of China (English)

    ZHANG YuGang; ZHANG LiDe

    2009-01-01

    ZnO nanostructures with three kinds of morphologies, namely, tetrapod-, rod-, and sheet-like shape, are synthesized by chemical vapor deposition, conventional solution-phase, and hydrothermal meth-ods, respectively. The fluorescence measurements display that the spectra of these nanostructures exhibit similar unexpected change laws with the altering excitation intensity. It is observed that when the excitation intensity increases, for the green emission band, the peak position shows a small blue-shift, the width turns broader, and the intensity grows first stronger and then weaker; for the UV emission band, the peak position exhibits a significant red-shift, and the width and intensity have the similar behaviors with those of the green band. Additionally, the relative intensity of green emission to UV emission decreases gradually. It is clarified that the origin of this abnormal phenomenon is as-cribed to the local laser heating effect and the high sensitivity of nanostructures to this heating effect.

  7. Effect of excitation intensity on fluorescence spectra in ZnO nanostructures and its origin

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    ZnO nanostructures with three kinds of morphologies, namely,tetrapod-,rod-,and sheet-like shape, are synthesized by chemical vapor deposition, conventional solution-phase,and hydrothermal methods, respectively.The fluorescence measurements display that the spectra of these nanostructures exhibit similar unexpected change laws with the altering excitation intensity. It is observed that when the excitation intensity increases, for the green emission band,the peak position shows a small blue-shift, the width turns broader,and the intensity grows first stronger and then weaker;for the UV emission band, the peak position exhibits a significant red-shift,and the width and intensity have the similar behaviors with those of the green band.Additionally,the relative intensity of green emission to UV emission decreases gradually.It is clarified that the origin of this abnormal phenomenon is ascribed to the local laser heating effect and the high sensitivity of nanostructures to this heating effect.

  8. [Fluorescence used to investigate the sensitivity of spinach chloroplast membrane to low intensity electromagnetic radiation].

    Science.gov (United States)

    Xi, Gang; Yang, Yun-Jing; Lu, Hong

    2009-07-01

    A system for studying biological effect of radio frequency electromagnetic field was developed. The system can form an area where electromagnetic wave with large frequency range is well distributed. The strength of electromagnetic wave was measured easily. Electromagnetic wave in the system did not have effect on environment. The sensitivity of spinach chloroplast membrane to low intensity electromagnetic radiation of 300 MHz under power density of 5 mW x cm(-2) was studied by the spectral analysis method of fluorescence of 8-anilino-1-naphthalene-sulfonic acid (ANS) and the changes in chlorophyll a (Chla) fluorescence parameters of spinach chloroplast membrane. The result showed that the position of spectrum of ANS fluorescence of spinach chloroplast membrane did not change, but the intensity of ANS fluorescence was obviously increased under the action of electromagnetic radiation with power density of 1-5 mW x cm(-2). There was an increase in the intensity of ANS fluorescence with the increase in electromagnetic radiation. The increase of ANS fluorescence of spinach chloroplast membrane showed that low level electromagnetic field induced the decrease in fluidity of chloroplast membrane compared with control experiment. The cause of the change in the fluidity could be related to the polarization of chloroplast membrane under the electromagnetic field. The analysis of Chla fluorescence parameters of spinach chloroplast membrane indicated that low level electromagnetic field of 300 MHz made the fluorescence parameters F0 and F(VI/)F(V) decrease, and F(V)/Fo, Fv/F(m) and deltaF(V)/T increase. It was showed that low level electromagnetic field caused the change of non-active center of photosystem II of spinach chloroplast membrane to active center and the increase in potential active and photochemical efficiency of PSII, and promoted the transmit process of electron in photosynthesis of chloroplast membrane of photosynthesis cell in spinach leaf. The study confirmed

  9. Effect of quencher and temperature on fluorescence intensity of laser dyes: DETC and C504T

    Science.gov (United States)

    Jana, Basavaraja; Inamdar, S. R.; Kumar, H. M. Suresh

    2017-01-01

    Fluorescence quenching of 7- Diethylamino-3-thenoylcoumarin (DETC) and 2,3,6,7-tetrahydro-1,1,7,7-tetramethyl11-oxo-1H,5H,11H- [1]benzopyrano[6,7,8-ij]quinolizine-10-carboxylic acid, ethyl ester (C504T) by aniline(AN), dimethylaniline (DMA) and diethylaniline (DEA) was investigated in toluene by steady state and transient methods. The quenching parameters like frequency of encounter (kd), probability of quenching per encounter (p), quenching rate parameters (kq) and activation energy of quenching (Ea) were determined experimentally. The kq values determined by steady state and time-resolved methods for the both dyes were found to be same, indicating the dynamic nature of interaction. Magnitudes of p and Ea suggested that the quenching reaction is predominantly controlled by material diffusion. The quenching mechanism is rationalized in terms of electron transfer (ET) from donors (aromatic amines) to the acceptors (coumarin derivatives) confirmed by correlating kq with free energy changes (ΔG°). Further, an effect of temperature on fluorescence intensity was carried out in toluene and methanol solvents. Fluorescence intensity of both the dyes decreases with increase in temperature. Temperature quenching in case of C504T is due to intersystem crossing S1 → T2, whereas for DETC, quenching is due to intersystem crossing S1 → T2 and ICT → TICT transition.

  10. Intense upconversion fluorescence in Tm 3+/Yb3+ codoped alumina lead borate glasses

    Science.gov (United States)

    Krishna Murthy Goud, K.; Shekhar Reddy, M. Chandra; Appa Rao, B.

    2016-09-01

    The Tm3+/Yb3+ codoped alumina lead borate glasses were prepared by the conventional melt quenching technique. Optical absorption and FTIR spectra were recorded. The upconversion fluorescence spectra exhibited weak blue (480 nm) and intense red (660 nm) emissions due to 1G4 → 3H6 and 1G4 → 3H4 transitions, respectively. The results concluded that both emissions are due to three photon absorption process. It has been observed that in the upconversion efficiency increases with the increase in the concentration of Yb3+ ions. The strong red upconversion fluorescence indicate that Tm3+/Yb3+ codoped alumina lead borate glasses can be used as potential host material for upconversion lasers.

  11. Analysis of Cell Movement by Simultaneous Quantification of Local Membrane Displacement and Fluorescent Intensities Using Quimp2

    NARCIS (Netherlands)

    Bosgraaf, Leonard; van Haastert, Peter J. M.; Bretschneider, Till

    The use of fluorescent markers in living cells has increased dramatically in the recent years. The quantitative analysis of the images requires specific analysis software. Previously, the program Quimp was launched for quantitating fluorescent intensities at the membrane or the cortex of the cell.

  12. Fluorescence intensity and lifetime-based cyanide sensitive probes for physiological safeguard

    Energy Technology Data Exchange (ETDEWEB)

    Badugu, Ramachandram [Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, Medical Biotechnology Center, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, MD 21201 (United States); Lakowicz, Joseph R. [Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, Medical Biotechnology Center, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, MD 21201 (United States)]. E-mail: lakowicz@cfs.umbi.umd.edu; Geddes, Chris D. [Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, Medical Biotechnology Center, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, MD 21201 (United States) and Institute of Fluorescence and Center for Fluorescence Spectroscopy, Medical Biotechnology Center, University of Maryland Biotechnology Institute, 725 West Lombard Street, Baltimore, MD 21201 (United States)]. E-mail: chris@cfs.umbi.umd.edu

    2004-09-20

    We characterize six new fluorescent probes that show both intensity and lifetime changes in the presence of free uncomplexed aqueous cyanide, allowing for fluorescence based cyanide sensing up to physiological safeguard levels, i.e. <30 {mu}M. One of the probes, m-BMQBA, shows a {approx}15-fold reduction in intensity and a {approx}10% change in mean lifetime at this level. The response of the new probes is based on their ability to bind the cyanide anion through a boronic acid functional group, changing from the neutral form of the boronic acid group R-B(OH){sub 2} to the anionic R-B{sup -}(CN){sub 3} form, a new cyanide binding mechanism which we have recently reported. The presence of an electron deficient quaternary heterocyclic nitrogen nucleus, and the electron rich cyanide bound form, provides for the intensity changes observed. We have determined the disassociation constants of the probes to be in the range {approx}15-84 {mu}M{sup 3}. In addition we have synthesized control compounds which do not contain the boronic acid moiety, allowing for a rationale of the cyanide responses between the probe isomers to be made. The lifetime of the cyanide bound probes are significantly shorter than the free R-B(OH){sub 2} probe forms, providing for the opportunity of lifetime based cyanide sensing up to physiologically lethal levels. Finally, while fluorescent probes containing the boronic acid moiety have earned a well-deserved reputation for monosaccharide sensing, we show that strong bases such as CN{sup -} and OH{sup -} preferentially bind as compared to glucose, enabling the potential use of these probes for cyanide safeguard and determination in physiological fluids, especially given that physiologies do not experience any notable changes in pH.

  13. Combining electrophoresis with detection under ultraviolet light and multiple ultrafiltration for isolation of humic fluorescence fractions.

    Science.gov (United States)

    Trubetskaya, Olga E; Shaloiko, Lubov A; Demin, Dmitrii V; Marchenkov, Victor V; Proskuryakov, Ivan I; Coelho, Christian; Trubetskoj, Oleg A

    2011-04-01

    Polyacrylamide gel electrophoresis of chernozem soil humic acids (HAs) followed by observation under UV (312 nm) excitation light reveals new low molecular weight (MW) fluorescent fractions. Ultrafiltration of HAs sample in 7 M urea on a membrane of low nominal MW retention (NMWR, 5 kDa) was repetitively used for separation of fluorescent and non-fluorescent species. Thirty ultrafiltrates and the final retentate R were obtained. Fluorescence maxima of separate ultrafiltrates were different and non-monotonously changed in the range of 475-505 nm. Fluorescence maxima of less than 490 nm were detected only in the four first utrafiltrates. For further physical-chemical analyses all utrafiltrates were combined into a fraction called UF<5 (NMW<5 kDa). Retentate R demonstrated very weak fluorescence under 270 nm excitation, while fluorescence intensity of UF<5 was about six times higher than of the bulk HAs. Fraction UF<5 was further ultrafiltrated on membranes of MNWR 3 kDa and 1 kDa, yielding three subfractions UF3-5, UF1-3 and UF<1 with NMW 3-5 kDa, 1-3 kDa and <1 kDa, respectively. The validation of the UF procedure was performed by size exclusion chromatography on Sephadex G-25 column. The fluorescence maxima were found to be at 505, 488 and 465 nm for UF3-5, UF1-3 and UF<1, respectively, with increasing of fluorescence intensity from UF3-5 to UF1-3 to UF<1 fraction. EPR analysis showed that the amount of free radicals was the largest in retentate R and drastically decreased in fluorescent ultrafiltrates. The results demonstrate that more than one fluorophore is present in chernozem soil HAs complex.

  14. Temperature Dependence of the Fluorescence Emission Intensity of Eu/TiO2 Nanocrystals

    Institute of Scientific and Technical Information of China (English)

    ZENG Qing-Guang; DING Ze-Jun; JU Xin; WANG Yi; SHENG Ye-Qing

    2007-01-01

    The samples of europium ions doped titanium dioxide (Eu3+/TiO2) nanocrystals are synthesized by a modified sol-gel method with hydrothermal treatment. The x-ray diffraction and scanning electron microscopy are used to characterize the sample. The temperature-dependent fluorescence emission effect of Eu3+ -doped samples is investigated. It is found that under the excitation of 514.5 nm light, the emission intensity of Eu3+ reaches a maximum value at 450K among various Eu3+ dopant concentrations in Eu3+/TiO2 nanocrystals. The variation of the emission intensity may be attributed to the photon-assist absorption and the temperature-quenching effect.

  15. [Intensity loss of two-photon excitation fluorescence microscopy images of mouse oocyte chromosomes].

    Science.gov (United States)

    Zhao, Feng-Ying; Wu, Hong-Xin; Chen, Die-Yan; Ma, Wan-Yun

    2014-07-01

    As an optical microscope with high resolution, two-photon excitation (TPE) fluorescence microscope is widely used in noninvasive 3D optical imaging of biological samples. Compared with confocal laser scanning microscope, TPE fluorescence microscope provides a deeper detecting depth. In spite of that, the image quality of sample always declines as the detecting depth increases when a noninvasive 3D optical imaging of thicker samples is performed. Mouse oocytes with a large diameter, which play an important role in clinical and biological fields, have obvious absorption and scattering effects. In the present paper, we performed compensation for two-photon fluorescence images of mouse oocyte chromosomes. Using volume as a parameter, the attenuation degree of these chromosomes was also studied. The result of our data suggested that there exists a severe axial intensity loss in two-photon microscopic images of mouse oocytes due to the absorption and scattering effects. It is necessary to make compensation for these images of mouse oocyte chromosomes obtained from two-photon microscopic system. It will be specially needed in studying the quantitative three-dimensional information of mouse oocytes.

  16. Quantum chemical calculations to reveal the relationship between the chemical structure and the fluorescence characteristics of phenylquinolinylethynes and phenylisoquinolinylethynes derivatives, and to predict their relative fluorescence intensity.

    Science.gov (United States)

    Riahi, Siavash; Beheshti, Abolghasem; Ganjali, Mohammad Reza; Norouzi, Parviz

    2009-12-01

    In this paper the relationship between the chemical structure and fluorescence characteristics of 30 phenylquinolinylethyne (PhQE), and phenylisoquinolinylethyne (PhIE) derivatives compounds employing ab initio calculations have been elucidated. Quantum chemical calculations (6-31G) were carried out to obtain: the optimized geometry, energy levels, charges and dipole moments of these compounds, in the singlet (steady and excited states) and triplet states. The relationship between quantum chemical descriptors, and wavelength of maximum excitation and emission indicated that these two parameters have the most correlation with quantum chemical hardness (eta). Also, stokes shift has the most correlation with the square of difference between the maximum of positive charges in the singlet steady and singlet excited states. The quantitative structure-property relationship (QSPR) of PhQE and PhIE was studied for relative fluorescence intensity (RFI). The genetic algorithm (GA) was applied to select the variables that resulted in the best-fit models. After the variable selection, multiple linear regression (MLR) and support vector machine (SVM) were both utilized to construct linear and non-linear QSPR models, respectively. The SVM model demonstrated a better performance than that of the MLR model. The route mean square error (RMSE) in the training and the test sets for the SVM model was 0.195 and 0.324, and the correlation coefficients were 0.965 and 0.960, respectively, thus revealing the reliability of this model. The resulting data indicated that SVM could be used as a powerful modeling tool for QSPR studies. According to the best of our knowledge, this is the first research on QSPR studies to predict RFI for a series of PhQE and PhIE derivative compounds using SVM.

  17. Mapping fast protein folding with multiple-site fluorescent probes.

    Science.gov (United States)

    Prigozhin, Maxim B; Chao, Shu-Han; Sukenik, Shahar; Pogorelov, Taras V; Gruebele, Martin

    2015-06-30

    Fast protein folding involves complex dynamics in many degrees of freedom, yet microsecond folding experiments provide only low-resolution structural information. We enhance the structural resolution of the five-helix bundle protein λ6-85 by engineering into it three fluorescent tryptophan-tyrosine contact probes. The probes report on distances between three different helix pairs: 1-2, 1-3, and 3-2. Temperature jump relaxation experiments on these three mutants reveal two different kinetic timescales: a slower timescale for 1-3 and a faster one for the two contacts involving helix 2. We hypothesize that these differences arise from a single folding mechanism that forms contacts on different timescales, and not from changes of mechanism due to adding the probes. To test this hypothesis, we analyzed the corresponding three distances in one published single-trajectory all-atom molecular-dynamics simulation of a similar mutant. Autocorrelation analysis of the trajectory reveals the same "slow" and "fast" distance change as does experiment, but on a faster timescale; smoothing the trajectory in time shows that this ordering is robust and persists into the microsecond folding timescale. Structural investigation of the all-atom computational data suggests that helix 2 misfolds to produce a short-lived off-pathway trap, in agreement with the experimental finding that the 1-2 and 3-2 distances involving helix 2 contacts form a kinetic grouping distinct from 1 to 3. Our work demonstrates that comparison between experiment and simulation can be extended to several order parameters, providing a stronger mechanistic test.

  18. Understanding the fundamental mechanisms of biofilms development and dispersal: BIAM (Biofilm Intensity and Architecture Measurement), a new tool for studying biofilms as a function of their architecture and fluorescence intensity.

    Science.gov (United States)

    Baudin, Marine; Cinquin, Bertrand; Sclavi, Bianca; Pareau, Dominique; Lopes, Filipa

    2017-09-01

    Confocal laser scanning microscopy (CLSM) is one of the most relevant technologies for studying biofilms in situ. Several tools have been developed to investigate and quantify the architecture of biofilms. However, an approach to quantify correctly the evolution of intensity of a fluorescent signal as a function of the structural parameters of a biofilm is still lacking. Here we present a tool developed in the ImageJ open source software that can be used to extract both structural and fluorescence intensity from CLSM data: BIAM (Biofilm Intensity and Architecture Measurement). This is of utmost significance when studying the fundamental mechanisms of biofilm growth, differentiation and development or when aiming to understand the effect of external molecules on biofilm phenotypes. In order to provide an example of the potential of such a tool in this study we focused on biofilm dispersion. cis-2-Decenoic acid (CDA) is a molecule known to induce biofilm dispersion of multiple bacterial species. The mechanisms by which CDA induces dispersion are still poorly understood. To investigate the effects of CDA on biofilms, we used a reporter strain of Escherichia coli (E. coli) that expresses the GFPmut2 protein under control of the rrnBP1 promoter. Experiments were done in flow cells and image acquisition was made with CLSM. Analysis carried out using the new tool, BIAM, indicates that CDA affects the fluorescence intensity of the biofilm structures as well as biofilm architectures. Indeed, our results demonstrate that CDA removes more than 35% of biofilm biovolume and suggest that it results in an increase of the biofilm's mean fluorescence intensity (MFI) by more than 26% compared to the control biofilm in the absence of CDA. Copyright © 2017. Published by Elsevier B.V.

  19. Comparative study of the fatty acid binding process of a new FABP from Cherax quadricarinatus by fluorescence intensity, lifetime and anisotropy.

    Directory of Open Access Journals (Sweden)

    Jiayao Li

    Full Text Available Fatty acid-binding proteins (FABPs are small cytosolic proteins, largely distributed in invertebrates and vertebrates, which accomplish uptake and intracellular transport of hydrophobic ligands such as fatty acids. Although long chain fatty acids play multiple crucial roles in cellular functions (structural, energy metabolism, regulation of gene expression, the precise functions of FABPs, especially those of invertebrate species, remain elusive. Here, we have identified and characterized a novel FABP family member, Cq-FABP, from the hepatopancreas of red claw crayfish Cherax quadricarinatus. We report the characterization of fatty acid-binding affinity of Cq-FABP by four different competitive fluorescence-based assays. In the two first approaches, the fluorescent probe 8-Anilino-1-naphthalenesulfonate (ANS, a binder of internal cavities of protein, was used either by directly monitoring its fluorescence emission or by monitoring the fluorescence resonance energy transfer occurring between the single tryptophan residue of Cq-FABP and ANS. The third and the fourth approaches were based on the measurement of the fluorescence emission intensity of the naturally fluorescent cis-parinaric acid probe or the steady-state fluorescence anisotropy measurements of a fluorescently labeled fatty acid (BODIPY-C16, respectively. The four methodologies displayed consistent equilibrium constants for a given fatty acid but were not equivalent in terms of analysis. Indeed, the two first methods were complicated by the existence of non specific binding modes of ANS while BODIPY-C16 and cis-parinaric acid specifically targeted the fatty acid binding site. We found a relationship between the affinity and the length of the carbon chain, with the highest affinity obtained for the shortest fatty acid, suggesting that steric effects primarily influence the interaction of fatty acids in the binding cavity of Cq-FABP. Moreover, our results show that the binding affinities

  20. Fluorescence Intensity and Polarization from Photodissociated H2, D2 and HD

    Science.gov (United States)

    Machacek, J. R.; Andrianarijaona, V. M.; Furst, J. E.; Gay, T. J.; Kilcoyne, A. L. D.; Landers, A. L.; McLaughlin, K. W.

    2009-05-01

    We have measured the intensity and linear polarization of Hα (n=3->n=2) 656.3 nm fluorescence resulting from H and D atoms created by photodissociation of H2, D2 and HD using linearly-polarized photons with energies ranging from 16.5 to 17.6 eV. Between the threshold for atomic n=3 production at 16.6 eV and the n=4 production threshold at 17.3 eV, the relative cross section and polarization data are free from cascade contributions due to higher-lying atomic states. The photon beam energy width used for this work was 3 meV. Comparison of relative intensities to previous measurements [1] show marked differences. However, the polarization is in qualitative agreement. [1] H. Frohlich et al., Z. Phys. D 34, 119 (1995). Support provided by the NSF (Grant PHY-0653379), DOE (LBNL/ALS) and ANSTO (Access to Major Research Facilities Programme).

  1. Differentiation of ocular fundus fluorophores by fluorescence lifetime imaging using multiple excitation and emission wavelengths

    Science.gov (United States)

    Hammer, M.; Schweitzer, D.; Schenke, S.; Becker, W.; Bergmann, A.

    2006-10-01

    Ocular fundus autofluorescence imaging has been introduced into clinical diagnostics recently. It is in use for the observation of the age pigment lipofuscin, a precursor of age - related macular degeneration (AMD). But other fluorophores may be of interest too: The redox pair FAD - FADH II provides information on the retinal energy metabolism, advanced glycation end products (AGE) indicate protein glycation associated with pathologic processes in diabetes as well as AMD, and alterations in the fluorescence of collagen and elastin in connective tissue give us the opportunity to observe fibrosis by fluorescence imaging. This, however, needs techniques able to differentiate particular fluorophores despite limited permissible ocular exposure as well as excitation wavelength (limited by the transmission of the human ocular lens to >400 nm). We present an ophthalmic laser scanning system (SLO), equipped with picosecond laser diodes (FWHM 100 ps, 446 nm or 468 nm respectively) and time correlated single photon counting (TCSPC) in two emission bands (500 - 560 nm and 560 - 700 nm). The decays were fitted by a bi-exponential model. Fluorescence spectra were measured by a fluorescence spectrometer fluorolog. Upon excitation at 446 nm, the fluorescence of AGE, FAD, and lipofuscin were found to peak at 503 nm, 525 nm, and 600 nm respectively. Accordingly, the statistical distribution of the fluorescence decay times was found to depend on the different excitation wavelengths and emission bands used. The use of multiple excitation and emission wavelengths in conjunction with fluorescence lifetime imaging allows us to discriminate between intrinsic fluorophores of the ocular fundus. Taken together with our knowledge on the anatomical structure of the fundus, these findings suggest an association of the short, middle and long fluorescence decay time to the retinal pigment epithelium, the retina, and connective tissue respectively.

  2. Effect of Host Medium on the Fluorescence Emission Intensity of Rhodamine B in Liquid and Solid Phase

    Science.gov (United States)

    Fikry, M.; Omar, M. M.; Ismail, Lotfi Z.

    2011-06-01

    In this work, we study the effect of concentration, host medium, PH, ions complex and phase states on the fluorescence emission from the laser dye, Rhodamine B, pumping by UV laser as exited source. The polymethylmethacrylate PMMA used as host medium in case of solid phase samples while, ethanol and Tetrahydrofuran (THF) are used in case of liquid one. The Laser Induced Fluorescence (LIF) technique was used to study the fluorescence properties of the both cases liquid and thin film solid-state samples. In addition, the Dual Thermal Lens (DTL) technique was used to study the quantum yield of these samples. The maximum fluorescence emission observed at concentration of Rhodamine B C=3×10-4M. At this concentration of Rhodamine B, the type of solvent and polarity of the medium affect on the fluorescence emission intensity of Rhodamine B with. The measurements revile that, the behavior of both phases state was analogous and Rhodamine B/PMMA thin film sample by ratio of 4:1 and thickness 0.12 mm is the best photostability sample and its quantum yield about ≈ 0.82. Also, the fluorescence emission intensity of Rhodamine B was quenched by complex formation of Co, Al, Cu and iodide ions with Rhodamine B due to the increase of the charge density of the ions.

  3. Amplification of the Signal Intensity of Fluorescence-Based Fiber-Optic Biosensors Using a Fabry-Perot Resonator Structure

    Directory of Open Access Journals (Sweden)

    Meng-Chang Hsieh

    2015-02-01

    Full Text Available Fluorescent biosensors have been widely used in biomedical applications. To amplify the intensity of fluorescence signals, this study developed a novel structure for an evanescent wave fiber-optic biosensor by using a Fabry-Perot resonator structure. An excitation light was coupled into the optical fiber through a laser-drilled hole on the proximal end of the resonator. After entering the resonator, the excitation light was reflected back and forth inside the resonator, thereby amplifying the intensity of the light in the fiber. Subsequently, the light was used to excite the fluorescent molecules in the reactive region of the sensor. The experimental results showed that the biosensor signal was amplified eight-fold when the resonator reflector was formed using a 92% reflective coating. Furthermore, in a simulation, the biosensor signal could be amplified 20-fold by using a 99% reflector.

  4. Resonance fluorescence of strongly driven two-level system coupled to multiple dissipative reservoirs

    Science.gov (United States)

    Yan, Yiying; Lü, Zhiguo; Zheng, Hang

    2016-08-01

    We present a theoretical formalism for resonance fluorescence radiating from a two-level system (TLS) driven by any periodic driving and coupled to multiple reservoirs. The formalism is derived analytically based on the combination of Floquet theory and Born-Markov master equation. The formalism allows us to calculate the spectrum when the Floquet states and quasienergies are analytically or numerically solved for simple or complicated driving fields. We can systematically explore the spectral features by implementing the present formalism. To exemplify this theory, we apply the unified formalism to comprehensively study a generic model that a harmonically driven TLS is simultaneously coupled to a radiative reservoir and a dephasing reservoir. We demonstrate that the significant features of the fluorescence spectra, the driving-induced asymmetry and the dephasing-induced asymmetry, can be attributed to the violation of detailed balance condition, and explained in terms of the driving-related transition quantities between Floquet-states and their steady populations. In addition, we find the distinguished features of the fluorescence spectra under the biharmonic and multiharmonic driving fields in contrast with that of the harmonic driving case. In the case of the biharmonic driving, we find that the spectra are significantly different from the result of the RWA under the multiple resonance conditions. By the three concrete applications, we illustrate that the present formalism provides a routine tool for comprehensively exploring the fluorescence spectrum of periodically strongly driven TLSs.

  5. Resonance fluorescence of strongly driven two-level system coupled to multiple dissipative reservoirs

    Energy Technology Data Exchange (ETDEWEB)

    Yan, Yiying, E-mail: yiyingyan@sjtu.edu.cn; Lü, Zhiguo, E-mail: zglv@sjtu.edu.cn; Zheng, Hang, E-mail: hzheng@sjtu.edu.cn

    2016-08-15

    We present a theoretical formalism for resonance fluorescence radiating from a two-level system (TLS) driven by any periodic driving and coupled to multiple reservoirs. The formalism is derived analytically based on the combination of Floquet theory and Born–Markov master equation. The formalism allows us to calculate the spectrum when the Floquet states and quasienergies are analytically or numerically solved for simple or complicated driving fields. We can systematically explore the spectral features by implementing the present formalism. To exemplify this theory, we apply the unified formalism to comprehensively study a generic model that a harmonically driven TLS is simultaneously coupled to a radiative reservoir and a dephasing reservoir. We demonstrate that the significant features of the fluorescence spectra, the driving-induced asymmetry and the dephasing-induced asymmetry, can be attributed to the violation of detailed balance condition, and explained in terms of the driving-related transition quantities between Floquet-states and their steady populations. In addition, we find the distinguished features of the fluorescence spectra under the biharmonic and multiharmonic driving fields in contrast with that of the harmonic driving case. In the case of the biharmonic driving, we find that the spectra are significantly different from the result of the RWA under the multiple resonance conditions. By the three concrete applications, we illustrate that the present formalism provides a routine tool for comprehensively exploring the fluorescence spectrum of periodically strongly driven TLSs.

  6. Three-dimensional fluorescent microscopy via simultaneous illumination and detection at multiple planes.

    Science.gov (United States)

    Ma, Qian; Khademhosseinieh, Bahar; Huang, Eric; Qian, Haoliang; Bakowski, Malina A; Troemel, Emily R; Liu, Zhaowei

    2016-08-16

    The conventional optical microscope is an inherently two-dimensional (2D) imaging tool. The objective lens, eyepiece and image sensor are all designed to capture light emitted from a 2D 'object plane'. Existing technologies, such as confocal or light sheet fluorescence microscopy have to utilize mechanical scanning, a time-multiplexing process, to capture a 3D image. In this paper, we present a 3D optical microscopy method based upon simultaneously illuminating and detecting multiple focal planes. This is implemented by adding two diffractive optical elements to modify the illumination and detection optics. We demonstrate that the image quality of this technique is comparable to conventional light sheet fluorescent microscopy with the advantage of the simultaneous imaging of multiple axial planes and reduced number of scans required to image the whole sample volume.

  7. Development of Pathological Diagnostics of Human Kidney Cancer by Multiple Staining Using New Fluorescent Fluolid Dyes

    Directory of Open Access Journals (Sweden)

    Dilibaier Wuxiuer

    2014-01-01

    Full Text Available New fluorescent Fluolid dyes have advantages over others such as stability against heat, dryness, and excess light. Here, we performed simultaneous immunostaining of renal tumors, clear cell renal cell carcinoma (RCC, papillary RCC, chromophobe RCC, acquired cystic disease-associated RCC (ACD-RCC, and renal angiomyolipoma (AML, with primary antibodies against Kank1, cytokeratin 7 (CK7, and CD10, which were detected with secondary antibodies labeled with Fluolid-Orange, Fluolid-Green, and Alexa Fluor 647, respectively. Kank1 was stained in normal renal tubules, papillary RCC, and ACD-RCC, and weakly or negatively in all other tumors. CK7 was positive in normal renal tubules, papillary RCC, and ACD-RCC. In contrast, CD10 was expressed in renal tubules and clear cell RCC, papillary RCC, AML, and AC-RCC, and weakly in chromophobe RCC. These results may contribute to differentiating renal tumors and subtypes of RCCs. We also examined the stability of fluorescence and found that fluorescent images of Fluolid dyes were identical between a tissue section and the same section after it was stored for almost three years at room temperature. This indicates that tissue sections can be stored at room temperature for a relatively long time after they are stained with multiple fluorescent markers, which could open a door for pathological diagnostics.

  8. Efficient HOMO-LUMO separation by multiple resonance effect toward ultrapure blue thermally activated delayed fluorescence

    Science.gov (United States)

    Hatakeyama, Takuji; Ikuta, Toshiaki; Shiren, Kazushi; Nakajima, Kiichi; Nomura, Shintaro; Ni, Jingping

    2016-09-01

    Organic light-emitting diodes (OLEDs) play an important role in the new generation of flat-panel displays. Conventional OLEDs employing fluorescent materials together with triplet-triplet annihilation suffer from a relatively low internal quantum efficiency (IQE) of 62.5%. On the other hand, the IQE of OLEDs employing phosphorescent or thermally activated delayed fluorescence (TADF) materials can reach 100%. However, these materials exhibit very broad peaks with a full-width at half-maximum (FWHM) of 70-100 nm and cannot satisfy the color-purity requirements for displays. Therefore, the latest commercial OLED displays employ blue fluorescent materials with a relatively low IQE, and efficient blue emitters with a small FWHM are highly needed. In our manuscript, we present organic molecules that exhibit ultrapure blue fluorescence based on TADF. These molecules consist of three benzene rings connected by one boron and two nitrogen atoms, which establish a rigid polycyclic framework and significant localization of the highest occupied and lowest unoccupied molecular orbitals by a multiple resonance effect. An OLED device based on the new emitter exhibits ultrapure blue emission at 467 nm with an FWHM of 28 nm, Commission Internationale de l'Eclairage (CIE) coordinates of (0.12, 0.13), and an IQE of 100%, which represent record-setting performance for blue OLED devices.

  9. Contribution of inner shell Compton ionization to the X-ray fluorescence line intensity

    Science.gov (United States)

    Fernández, Jorge E.; Scot, Viviana; Di Giulio, Eugenio

    2016-10-01

    The Compton effect is a potential ionization mechanism of atoms. It produces vacancies in inner shells that are filled with the same mechanism of atomic relaxation as the one following photo-absorption. This contribution to X-ray fluorescence emission is frequently neglected because the total Compton cross-section is apparently much lower than the photoelectric one at useful X-ray energies. However, a more careful analysis suggests that is necessary to consider single shell cross sections (instead of total cross sections) as a function of energy. In this article these Compton cross sections are computed for the shells K, L1-L3 and M1-M5 in the framework of the impulse approximation. By comparing the Compton and the photoelectric cross-section for each shell it is then possible to determine the extent of the Compton correction to the intensity of the corresponding characteristic lines. It is shown that for the K shell the correction becomes relevant for excitation energies which are too high to be influent in X-ray spectrometry. In contrast, for L and M shells the Compton contribution is relevant for medium-Z elements and medium energies. To illustrate the different grades of relevance of the correction, for each ionized shell, the energies for which the Compton contribution reaches the extent levels of 1, 5, 10, 20, 50 and 100% of the photoelectric one are determined for all the elements with Z = 11-92. For practical applications it is provided a simple formula and fitting coefficients to compute average correction levels for the shells considered.

  10. Spatial dynamics of laser-induced fluorescence in an intense laser beam: experiment and theory in alkali metal atoms

    CERN Document Server

    Auzinsh, Marcis; Ferber, Ruvin; Gahbauer, Florian; Kalnins, Uldis

    2015-01-01

    We have shown that it is possible to model accurately optical phenomena in intense laser fields by taking into account the intensity distribution over the laser beam. We developed a theoretical model that divided an intense laser beam into concentric regions, each with a Rabi frequency that corresponds to the intensity in that region, and solved a set of coupled optical Bloch equations for the density matrix in each region. Experimentally obtained magneto-optical resonance curves for the $F_g=2\\longrightarrow F_e=1$ transition of the $D_1$ line of $^{87}$Rb agreed very well with the theoretical model up to a laser intensity of around 200 mW/cm$^2$ for a transition whose saturation intensity is around 4.5 mW/cm$^2$. We have studied the spatial dependence of the fluorescence intensity in an intense laser beam experimentally and theoretically. An experiment was conducted whereby a broad, intense pump laser excited the $F_g=4\\longrightarrow F_e=3$ transition of the $D_2$ line of cesium while a weak, narrow probe ...

  11. Cooperative effects and slow dynamics of fluorescence intensity from quantum emitters in a dielectric

    Science.gov (United States)

    Lozing, N. A.; Gladush, M. G.

    2016-12-01

    We study theoretically the possibility of spontaneous switching between dim and bright fluorescence modes from a cooperative ensemble of two-level atoms driven by a cw-laser. A numerical analysis of transient regimes and transformations of the fluorescence spectrum are reported.

  12. Oxygen evolution and chlorophyll fluorescence from multiple turnover light pulses: charge recombination in photosystem II in sunflower leaves.

    Science.gov (United States)

    Laisk, Agu; Oja, Vello; Eichelmann, Hillar

    2012-09-01

    Oxygen evolution and Chl fluorescence induction were measured during multiple turnover light pulses (MTP) of 630-nm wavelength, intensities from 250 to 8,000 μmol quanta m(-2) s(-1) and duration from 0.3 to 200 ms in sunflower leaves at 22 °C. The ambient O(2) concentration was 10-30 ppm and MTP were applied after pre-illumination under far-red light (FRL), which oxidized plastoquinone (PQ) and randomized S-states because of the partial excitation of PSII. Electron (e ( - )) flow was calculated as 4·O(2) evolution. Illumination with MTP of increasing length resulted in increasing O(2) evolution per pulse, which was differentiated against pulse length to find the time course of O(2) evolution rate with sub-millisecond resolution. Comparison of the quantum yields, Y (IIO) = e ( - )/hν from O(2) evolution and Y (IIF) = (F (m) - F)/F (m) from Chl fluorescence, detected significant losses not accompanied by fluorescence emission. These quantum losses are discussed to be caused by charge recombination between Q (A) (-) and oxidized TyrZ at a rate of about 1,000 s(-1), either directly or via the donor side equilibrium complex Q(A) → P (D1) (+)  ↔ TyrZ(ox), or because of cycling facilitated by Cyt b (559). Predicted from the suggested mechanism, charge recombination is enhanced by damage to the water-oxidizing complex and by restricted PSII acceptor side oxidation. The rate of PSII charge recombination/cycling is fast enough for being important in photoprotection.

  13. Chemical Environment Effects on K[beta]/K[alpha] Intensity Ratio: An X-Ray Fluorescence Experiment on Periodic Trends

    Science.gov (United States)

    Durham, Chaney R.; Chase, Jeffery M.; Nivens, Delana A.; Baird, William H.; Padgett, Clifford W.

    2011-01-01

    X-ray fluorescence (XRF) data from an energy-dispersive XRF instrument were used to investigate the chlorine K[alpha] and K[beta] peaks in several group 1 salts. The ratio of the peak intensity is sensitive to the local chemical environment of the chlorine atoms studied in this experiment and it shows a periodic trend for these salts. (Contains 1…

  14. Study on the Intensive Use of Rural Cemetery Lands from the Perspective of Multiple Functions

    Institute of Scientific and Technical Information of China (English)

    Li; Mingping; Yu; Zhongxiang

    2014-01-01

    According to the investigation and study on the land use of different cemeteries in rural areas,the intensive land use of four cemeteries,including the public cemetery,traditional cemetery,vertical cemetery,and ecological cemetery,was evaluated.According to the data analysis,it is suggested to choose right cemetery according the practical conditions of each area,to take full use the multiple functions of cemetery lands and improve the intensive use.

  15. Generation of broadband spontaneous parametric fluorescence using multiple bulk nonlinear crystals

    CERN Document Server

    Okano, Masayuki; Tanaka, Akira; Subashchandran, Shanthi; Takeuchi, Shigeki; 10.1364/OE.20.013977

    2012-01-01

    We propose a novel method for generating broadband spontaneous parametric fluorescence by using a set of bulk nonlinear crystals (NLCs). We also demonstrate this scheme experimentally. Our method employs a superposition of spontaneous parametric fluorescence spectra generated using multiple bulk NLCs. A typical bandwidth of 160 nm (73 THz) with a degenerate wavelength of 808 nm was achieved using two beta-barium-borate (BBO) crystals, whereas a typical bandwidth of 75 nm (34 THz) was realized using a single BBO crystal. We also observed coincidence counts of generated photon pairs in a non-collinear configuration. The bandwidth could be further broadened by increasing the number of NLCs. Our demonstration suggests that a set of four BBO crystals could realize a bandwidth of approximately 215 nm (100 THz).We also discuss the stability of Hong-Ou-Mandel two-photon interference between the parametric fluorescence generated by this scheme. Our simple scheme is easy to implement with conventional NLCs and does not...

  16. Generation of broadband spontaneous parametric fluorescence using multiple bulk nonlinear crystals.

    Science.gov (United States)

    Okano, Masayuki; Okamoto, Ryo; Tanaka, Akira; Subashchandran, Shanthi; Takeuchi, Shigeki

    2012-06-18

    We propose a novel method for generating broadband spontaneous parametric fluorescence by using a set of bulk nonlinear crystals (NLCs). We also demonstrate this scheme experimentally. Our method employs a superposition of spontaneous parametric fluorescence spectra generated using multiple bulk NLCs. A typical bandwidth of 160 nm (73 THz) with a degenerate wavelength of 808 nm was achieved using two β-barium-borate (BBO) crystals, whereas a typical bandwidth of 75 nm (34 THz) was realized using a single BBO crystal. We also observed coincidence counts of generated photon pairs in a non-collinear configuration. The bandwidth could be further broadened by increasing the number of NLCs. Our demonstration suggests that a set of four BBO crystals could realize a bandwidth of approximately 215 nm (100 THz). We also discuss the stability of Hong-Ou-Mandel two-photon interference between the parametric fluorescence generated by this scheme. Our simple scheme is easy to implement with conventional NLCs and does not require special devices.

  17. Effects of water stress and light intensity on chlorophyll fluorescence parameters and pigments of Aloe vera L.

    Science.gov (United States)

    Hazrati, Saeid; Tahmasebi-Sarvestani, Zeinolabedin; Modarres-Sanavy, Seyed Ali Mohammad; Mokhtassi-Bidgoli, Ali; Nicola, Silvana

    2016-09-01

    Aloe vera L. is one of the most important medicinal plants in the world. In order to determine the effects of light intensity and water deficit stress on chlorophyll (Chl) fluorescence and pigments of A. vera, a split-plot in time experiment was laid out in a randomized complete block design with four replications in a research greenhouse. The factorial combination of three light intensities (50, 75 and 100% of sunlight) and four irrigation regimes (irrigation after depleting 20, 40, 60 and 80% of soil water content) were considered as main factors. Sampling time was considered as sub factor. The first, second and third samplings were performed 90, 180 and 270 days after imposing the treatments, respectively. The results demonstrated that the highest light intensity and the severe water stress decreased maximum fluorescence (Fm), variable fluorescence (Fv)/Fm, quantum yield of PSII photochemistry (ФPSII), Chl and photochemical quenching (qP) but increased non-photochemical quenching (NPQ), minimum fluorescence (F0) and Anthocyanin (Anth). Additionally, the highest Fm, Fv/Fm, ФPSII and qP and the lowest NPQ and F0 were observed when 50% of sunlight was blocked and irrigation was done after 40% soil water depletion. Irradiance of full sunlight and water deficit stress let to the photoinhibition of photosynthesis, as indicated by a reduced quantum yield of PSII, ФPSII, and qP, as well as higher NPQ. Thus, chlorophyll florescence measurements provide valuable physiological data. Close to half of total solar radiation and irrigation after depleting 40% of soil water content were selected as the most efficient treatments.

  18. Non-destructive mobile monitoring of microbial contaminations on meat surfaces using porphyrin fluorescence intensities.

    Science.gov (United States)

    Durek, J; Fröhling, A; Bolling, J; Thomasius, R; Durek, P; Schlüter, O K

    2016-05-01

    A non-destructive mobile system for meat quality monitoring was developed and investigated for the possible application along the whole production chain of fresh meat. Pork and lamb meat was stored at 5 °C for up to 20 days post mortem and measured with a fluorescence spectrometer. Additionally, the bacterial influence on the fluorescence signals was evaluated by different experimental procedures. Fluorescence of NADH and different porphyrins could be correlated to the growth of diverse bacteria and hence used for contamination monitoring. The increase of porphyrin fluorescence started after 9 days p.m. for pork and after 2 days p.m. for lamb meat. Based on the results, a mobile fluorescence system was built and compared with the laboratory system. The corrected function of the meat slices showed a root mean square error of 1156.97 r.u. and a mean absolute percentage error of 12.59%; for lamb the values were 470.81 r.u. and 15.55%, respectively. A mobile and non-invasive measurement system would improve the microbial security of fresh meat.

  19. Intense Internal and External Fluorescence as Solar Cells Approach the Shockley-Queisser Efficiency Limit

    CERN Document Server

    Miller, Owen D; Kurtz, Sarah R

    2012-01-01

    Absorbed sunlight in a solar cell produces electrons and holes. But, at the open circuit condition, the carriers have no place to go. They build up in density and, ideally, they emit external fluorescence that exactly balances the incoming sunlight. Any additional non-radiative recombination impairs the carrier density buildup, limiting the open-circuit voltage. At open-circuit, efficient external fluorescence is an indicator of low internal optical losses. Thus efficient external fluorescence is, counter-intuitively, a necessity for approaching the Shockley-Queisser efficiency limit. A great Solar Cell also needs to be a great Light Emitting Diode. Owing to the narrow escape cone for light, efficient external emission requires repeated attempts, and demands an internal luminescence efficiency >>90%.

  20. Multiple functionalization of fluorescent nanoparticles for specific biolabeling and drug delivery of dopamine

    Science.gov (United States)

    Malvindi, Maria Ada; di Corato, Riccardo; Curcio, Annalisa; Melisi, Daniela; Rimoli, Maria Grazia; Tortiglione, Claudia; Tino, Angela; George, Chandramohan; Brunetti, Virgilio; Cingolani, Roberto; Pellegrino, Teresa; Ragusa, Andrea

    2011-12-01

    The development of fluorescent biolabels for specific targeting and controlled drug release is of paramount importance in biological applications due to their potential in the generation of novel tools for simultaneous diagnosis and treatment of diseases. Dopamine is a neurotransmitter involved in several neurological diseases, such as Parkinson's disease and attention deficit hyperactivity disorder (ADHD), and the controlled delivery of its agonists already proved to have beneficial effects both in vitro and in vivo. Here, we report the synthesis and multiple functionalization of highly fluorescent CdSe/CdS quantum rods for specific biolabeling and controlled drug release. After being transferred into aqueous media, the nanocrystals were made highly biocompatible through PEG conjugation and covered by a carbohydrate shell, which allowed specific GLUT-1 recognition. Controlled attachment of dopamine through an ester bond also allowed hydrolysis by esterases, yielding a smart nanotool for specific biolabeling and controlled drug release.The development of fluorescent biolabels for specific targeting and controlled drug release is of paramount importance in biological applications due to their potential in the generation of novel tools for simultaneous diagnosis and treatment of diseases. Dopamine is a neurotransmitter involved in several neurological diseases, such as Parkinson's disease and attention deficit hyperactivity disorder (ADHD), and the controlled delivery of its agonists already proved to have beneficial effects both in vitro and in vivo. Here, we report the synthesis and multiple functionalization of highly fluorescent CdSe/CdS quantum rods for specific biolabeling and controlled drug release. After being transferred into aqueous media, the nanocrystals were made highly biocompatible through PEG conjugation and covered by a carbohydrate shell, which allowed specific GLUT-1 recognition. Controlled attachment of dopamine through an ester bond also allowed

  1. Hoechst fluorescence intensity can be used to separate viable bromodeoxyuridine-labeled cells from viable non-bromodeoxyuridine-labeled cells

    Science.gov (United States)

    Mozdziak, P. E.; Pulvermacher, P. M.; Schultz, E.; Schell, K.

    2000-01-01

    BACKGROUND: 5-Bromo-2'-deoxyuridine (BrdU) is a powerful compound to study the mitotic activity of a cell. Most techniques that identify BrdU-labeled cells require conditions that kill the cells. However, the fluorescence intensity of the membrane-permeable Hoechst dyes is reduced by the incorporation of BrdU into DNA, allowing the separation of viable BrdU positive (BrdU+) cells from viable BrdU negative (BrdU-) cells. METHODS: Cultures of proliferating cells were supplemented with BrdU for 48 h and other cultures of proliferating cells were maintained without BrdU. Mixtures of viable BrdU+ and viable BrdU- cells from the two proliferating cultures were stained with Hoechst 33342. The viable BrdU+ and BrdU- cells were sorted into different fractions from a mixture of BrdU+ and BrdU- cells based on Hoechst fluorescence intensity and the ability to exclude the vital dye, propidium iodide. Subsequently, samples from the original mixture, the sorted BrdU+ cell population, and the sorted BrdU- cell population were immunostained using an anti-BrdU monoclonal antibody and evaluated using flow cytometry. RESULTS: Two mixtures consisting of approximately 55% and 69% BrdU+ cells were sorted into fractions consisting of greater than 93% BrdU+ cells and 92% BrdU- cells. The separated cell populations were maintained in vitro after sorting to demonstrate their viability. CONCLUSIONS: Hoechst fluorescence intensity in combination with cell sorting is an effective tool to separate viable BrdU+ from viable BrdU- cells for further study. The separated cell populations were maintained in vitro after sorting to demonstrate their viability. Copyright 2000 Wiley-Liss, Inc.

  2. Multiple Signal Classification Algorithm (MUSICAL) for super-resolution fluorescence microscopy

    CERN Document Server

    Agarwal, Krishna

    2016-01-01

    Super-resolution microscopy is providing unprecedented insights into biology by resolving details much below the diffraction limit. State-of-the-art Single Molecule Localization Microscopy (SMLM) techniques for super-resolution are restricted by long acquisition and computational times, or the need of special fluorophores or chemical environments. Here, we propose a novel statistical super-resolution technique of wide-field fluorescence microscopy called MUltiple SIgnal Classification ALgorithm (MUSICAL) which has several advantages over SMLM techniques. MUSICAL provides resolution down to at least 50 nm, has low requirements on number of frames and excitation power and works even at high fluorophore concentrations. Further, it works with any fluorophore that exhibits blinking on the time scale of the recording. We compare imaging results of MUSICAL with SMLM and four contemporary statistical super-resolution methods for experiments of in-vitro actin filaments and datasets provided by independent research gro...

  3. Variational level set approach for automatic correction of multiplicative and additive intensity inhomogeneities in brain MR Images.

    Science.gov (United States)

    Verma, Nishant; Cowperthwaite, Matthew C; Markey, Mia K

    2012-01-01

    Retrospective correction of intensity inhomogeneities in magnetic resonance images of the brain is an essential pre-processing step before any sophisticated image analysis task can be performed. A popular choice when defining the degradation model in MR images is to use multiplicative intensity inhomogeneities that slowly varying across the image domain; this approach has been extensively used for bias field estimation. However, such a multiplicative model is often insufficient given that some of the most dominant physical causes of intensity inhomogeneities in MRI (such as nonuniform excitation strength) have a non-linear relationship with the receptor signal intensity. In this study, we consider a linear image degradation model with multiplicative and additive intensity inhomogeneity components. We propose a variational level sets approach that combines estimation of intensity inhomogeneity components during the image segmentation process. The evaluation of proposed approach on real MR image datasets demonstrate accurate estimation of multiplicative and additive intensity inhomogeneities improving brain tissue segmentation.

  4. Combining OCT and a fluorescence intensity imaging method for atherosclerosis detection

    Science.gov (United States)

    Liang, Shanshan; Saidi, Arya; Jing, Joe; Liu, Gangjun; Yin, Jiechen; Narula, Jagat; Chen, Zhongping

    2012-02-01

    Coronary heart disease (like myocardial infarction) is caused by atherosclerosis. It cause over 30% of all deaths in North America and are the most common cause of death in European men under 65 years of age and the second most common cause in women. To diagnose this atherosclerosis before it gets rupture is the most effect way to increase the chance of survival for patients who suffer from this disease. The crucial tusk is how to find out vulnerable plaques. In resent years optical coherence tomography (OCT) has become a very useful tool for intravascular imaging, since it has high axial and transverse resolution. OCT can tell the detail structure inside the plaque like the thickness of plaque cap which is an important factor to identify vulnerable plaques. But we still need to find out the biochemical characteristics that is unique for vulnerable plaques (like inflammation). Fluorescence molecular imaging is a standard way to exam the biochemical property of biological samples. So we integrate these two techniques together into one probe. Our probe is comprised of a double-clad fiber (DCF) and a grin lens, and rotates with a micro mirror in front. The single-mode inner core of the DCF transmits both OCT and fluorescence excitation light, and the multimode inner cladding is used to detect fluorescence signal. In vitro result shows that this is a possible way for more accurate diagnose of vulnerable plaques.

  5. Surface Coverage and Structure of Mixed DNA/Alkylthiol Monolayers on Gold: Characterization by XPS, NEXAFS, and Fluorescence Intensity Measurements

    Energy Technology Data Exchange (ETDEWEB)

    Lee,C.; Gong, P.; Harbers, G.; Grainger, D.; Castner, D.; Gamble, L.

    2006-01-01

    Self-assembly of thiol-terminated single-stranded DNA (HS-ssDNA) on gold has served as an important model system for DNA immobilization at surfaces. Here, we report a detailed study of the surface composition and structure of mixed self-assembled DNA monolayers containing a short alkylthiol surface diluent [11-mercapto-1-undecanol (MCU)] on gold supports. These mixed DNA monolayers were studied with X-ray photoelectron spectroscopy (XPS), near-edge X-ray absorption fine structure spectroscopy (NEXAFS), and fluorescence intensity measurements. XPS results on sequentially adsorbed DNA/MCU monolayers on gold indicated that adsorbed MCU molecules first incorporate into the HS-ssDNA monolayer and, upon longer MCU exposures, displace adsorbed HS-ssDNA molecules from the surface. Thus, HS-ssDNA surface coverage steadily decreased with MCU exposure time. Polarization-dependent NEXAFS and fluorescence results both show changes in signals consistent with changes in DNA orientation after only 30 min of MCU exposure. NEXAFS polarization dependence (followed by monitoring the N 1s{yields}{pi}* transition) of the mixed DNA monolayers indicated that the DNA nucleotide base ring structures are oriented more parallel to the gold surface compared to DNA bases in pure HS-ssDNA monolayers. This indicates that HS-ssDNA oligomers reorient toward a more-upright position upon MCU incorporation. Fluorescence intensity results using end-labeled DNA probes on gold show little observable fluorescence on pure HS-ssDNA monolayers, likely due to substrate quenching effects between the fluorophore and the gold. MCU diluent incorporation into HS-ssDNA monolayers initially increases DNA fluorescence signal by densifying the chemisorbed monolayer, prompting an upright orientation of the DNA, and moving the terminal fluorophore away from the substrate. Immobilized DNA probe density and DNA target hybridization in these mixed DNA monolayers, as well as effects of MCU diluent on DNA hybridization in

  6. Surface Coverage and Structure of Mixed DNA/Alkylthiol Monolayers on Gold: Characterization by XPS, NEXAFS, and Fluorescence Intensity Measurements

    Science.gov (United States)

    Lee, Chi-Ying; Gong, Ping; Harbers, Gregory M.; Grainger, David W.; Castner, David G.; Gamble, Lara J.

    2006-01-01

    Self-assembly of thiol-terminated single-stranded DNA (HS-ssDNA) on gold has served as an important model system for DNA immobilization at surfaces. Here, we report a detailed study of the surface composition and structure of mixed self-assembled DNA monolayers containing a short alkylthiol surface diluent [11-mercapto-1-undecanol (MCU)] on gold supports. These mixed DNA monolayers were studied with X-ray photoelectron spectroscopy (XPS), near-edge X-ray absorption fine structure spectroscopy (NEXAFS), and fluorescence intensity measurements. XPS results on sequentially adsorbed DNA/MCU monolayers on gold indicated that adsorbed MCU molecules first incorporate into the HS-ssDNA monolayer and, upon longer MCU exposures, displace adsorbed HS-ssDNA molecules from the surface. Thus, HS-ssDNA surface coverage steadily decreased with MCU exposure time. Polarization-dependent NEXAFS and fluorescence results both show changes in signals consistent with changes in DNA orientation after only 30 min of MCU exposure. NEXAFS polarization dependence (followed by monitoring the N 1s → π* transition) of the mixed DNA monolayers indicated that the DNA nucleotide base ring structures are oriented more parallel to the gold surface compared to DNA bases in pure HS-ssDNA monolayers. This indicates that HS-ssDNA oligomers reorient toward a more-upright position upon MCU incorporation. Fluorescence intensity results using end-labeled DNA probes on gold show little observable fluorescence on pure HS-ssDNA monolayers, likely due to substrate quenching effects between the fluorophore and the gold. MCU diluent incorporation into HS-ssDNA monolayers initially increases DNA fluorescence signal by densifying the chemisorbed monolayer, prompting an upright orientation of the DNA, and moving the terminal fluorophore away from the substrate. Immobilized DNA probe density and DNA target hybridization in these mixed DNA monolayers, as well as effects of MCU diluent on DNA hybridization in complex

  7. Surface coverage and structure of mixed DNA/alkylthiol monolayers on gold: characterization by XPS, NEXAFS, and fluorescence intensity measurements.

    Science.gov (United States)

    Lee, Chi-Ying; Gong, Ping; Harbers, Gregory M; Grainger, David W; Castner, David G; Gamble, Lara J

    2006-05-15

    Self-assembly of thiol-terminated single-stranded DNA (HS-ssDNA) on gold has served as an important model system for DNA immobilization at surfaces. Here, we report a detailed study of the surface composition and structure of mixed self-assembled DNA monolayers containing a short alkylthiol surface diluent [11-mercapto-1-undecanol (MCU)] on gold supports. These mixed DNA monolayers were studied with X-ray photoelectron spectroscopy (XPS), near-edge X-ray absorption fine structure spectroscopy (NEXAFS), and fluorescence intensity measurements. XPS results on sequentially adsorbed DNA/MCU monolayers on gold indicated that adsorbed MCU molecules first incorporate into the HS-ssDNA monolayer and, upon longer MCU exposures, displace adsorbed HS-ssDNA molecules from the surface. Thus, HS-ssDNA surface coverage steadily decreased with MCU exposure time. Polarization-dependent NEXAFS and fluorescence results both show changes in signals consistent with changes in DNA orientation after only 30 min of MCU exposure. NEXAFS polarization dependence (followed by monitoring the N 1s --> pi* transition) of the mixed DNA monolayers indicated that the DNA nucleotide base ring structures are oriented more parallel to the gold surface compared to DNA bases in pure HS-ssDNA monolayers. This indicates that HS-ssDNA oligomers reorient toward a more-upright position upon MCU incorporation. Fluorescence intensity results using end-labeled DNA probes on gold show little observable fluorescence on pure HS-ssDNA monolayers, likely due to substrate quenching effects between the fluorophore and the gold. MCU diluent incorporation into HS-ssDNA monolayers initially increases DNA fluorescence signal by densifying the chemisorbed monolayer, prompting an upright orientation of the DNA, and moving the terminal fluorophore away from the substrate. Immobilized DNA probe density and DNA target hybridization in these mixed DNA monolayers, as well as effects of MCU diluent on DNA hybridization in complex

  8. Correlation between the In Vitro Functionality of Stored Platelets and the Cytosolic Esterase-Induced Fluorescence Intensity with CMFDA.

    Science.gov (United States)

    Wang, Jiexi; Yi, Xiaoyang; Liu, Minxia; Zhou, Qian; Ren, Suping; Wang, Yan; Yang, Chao; Zhou, Jianwei; Han, Ying

    2015-01-01

    It has been hypothesized that the cytosolic esterase-induced fluorescence intensity (CEIFI) from carboxy dimethyl fluorescein diacetate (CMFDA) in platelets may related to platelet functions. In the present study, we measured the change of CEIFI in platelets during storage, and examined the correlations of CEIFI with the in vitro functionality of stored platelets, including the ADP-induced aggregation activity, hypotonic shock response, expression of CD62P as well as platelet apoptosis. The CEIFI of fresh platelets, when tested at 10 μM CMFDA, the mean fluorescence intensity index (MFI) was 305.9 ± 49.9 (N = 80). After 1-day storage, it was 203.8 ± 34.4, the CEIFI of the stored platelets started to decline significantly, and reduced to 112.7 ±27.7 after 7-day storage. The change in CEIFI is highly correlated to all four functional parameters measured, with the correlation coefficients being 0.9813, 0.9848, -0.9945 and -0.9847 for the ADP-induced aggregation activity, hypotonic shock response (HSR), expression of CD62P and platelet apoptosis respectively. The above results show that the CEIFI measurement of platelets represents well the viability and functional state of in vitro stored platelets. This may be used as a convenient new method for quality evaluation for stored platelets if this result can be further validated by the following clinical trials.

  9. Characterization and application of PBA fiber optic chemical film sensor based on fluorescence multiple quenching

    Institute of Scientific and Technical Information of China (English)

    陈坚; 李伟; 阎超; 袁立懋; 郭炬亮; 周新继

    1997-01-01

    The three types of structure of the pyrenebutyric acid of fiber optic chemical film sensor were stud-ied by fluorescence multiple quenching. They are, for different test samples and purposes, respectively general, three-way and combined. A tri-cup method was designed to demonstrate the multiple quenching of response mechanism, and a relationship formula of mathematical approach was established. The response mechanism was shown to include the dynamic quenching , inner-filter effects and/or resonance energy transfer. To show the response characterization in a series of organic and inorganic quenchers, a new concept of apparent quenching coefficient Kq was advanced. This kind of sensor has been used in continuous and in situ monitoring of the dissolution rate of drug tablets, on line and in situ monitoring of some organic therapeutic drugs in biological fluid and Cr( VI ) in industrial waste water. The measured data were examined and compared with HPLC or HPTLCS. Test results show that the sensors and appa

  10. A fluorescent cassette-based strategy for engineering multiple domain fusion proteins

    Directory of Open Access Journals (Sweden)

    Khorchid Ahmad

    2003-07-01

    Full Text Available Abstract Background The engineering of fusion proteins has become increasingly important and most recently has formed the basis of many biosensors, protein purification systems, and classes of new drugs. Currently, most fusion proteins consist of three or fewer domains, however, more sophisticated designs could easily involve three or more domains. Using traditional subcloning strategies, this requires micromanagement of restriction enzymes sites that results in complex workaround solutions, if any at all. Results Therefore, to aid in the efficient construction of fusion proteins involving multiple domains, we have created a new expression vector that allows us to rapidly generate a library of cassettes. Cassettes have a standard vector structure based on four specific restriction endonuclease sites and using a subtle property of blunt or compatible cohesive end restriction enzymes, they can be fused in any order and number of times. Furthermore, the insertion of PCR products into our expression vector or the recombination of cassettes can be dramatically simplified by screening for the presence or absence of fluorescence. Conclusions Finally, the utility of this new strategy was demonstrated by the creation of basic cassettes for protein targeting to subcellular organelles and for protein purification using multiple affinity tags.

  11. Analysis of simulated fluorescence intensities decays by a new maximum entropy method algorithm.

    Science.gov (United States)

    Esposito, Rosario; Altucci, Carlo; Velotta, Raffaele

    2013-01-01

    A new algorithm for the Maximum Entropy Method (MEM) is proposed for recovering the lifetime distribution in time-resolved fluorescence decays. The procedure is based on seeking the distribution that maximizes the Skilling entropy function subjected to the chi-squared constraint χ(2) ~ 1 through iterative linear approximations, LU decomposition of the Hessian matrix of the lagrangian problem and the Golden Section Search for backtracking. The accuracy of this algorithm has been investigated through comparisons with simulated fluorescence decays both of narrow and broad lifetime distributions. The proposed approach is capable to analyse datasets of up to 4,096 points with a discretization ranging from 100 to 1,000 lifetimes. A good agreement with non linear fitting estimates has been observed when the method has been applied to multi-exponential decays. Remarkable results have been also obtained for the broad lifetime distributions where the position is recovered with high accuracy and the distribution width is estimated within 3%. These results indicate that the procedure proposed generates MEM lifetime distributions that can be used to quantify the real heterogeneity of lifetimes in a sample.

  12. Optimal Gaussian Mixture Models of Tissue Intensities in Brain MRI of Patients with Multiple-Sclerosis

    Science.gov (United States)

    Xiao, Yiming; Shah, Mohak; Francis, Simon; Arnold, Douglas L.; Arbel, Tal; Collins, D. Louis

    Brain tissue segmentation is important in studying markers in human brain Magnetic Resonance Images (MRI) of patients with diseases such as Multiple Sclerosis (MS). Parametric segmentation approaches typically assume unimodal Gaussian distributions on MRI intensities of individual tissue classes, even in applications on multi-spectral images. However, this assumption has not been rigorously verified especially in the context of MS. In this work, we evaluate the local MRI intensities of both healthy and diseased brain tissues of 21 multi-spectral MRIs (63 volumes in total) of MS patients for adherence to this assumption. We show that the tissue intensities are not uniform across the brain and vary across (anatomical) regions of the brain. Consequently, we show that Gaussian mixtures can better model the multi-spectral intensities. We utilize an Expectation Maximization (EM) based approach to learn the models along with a symmetric Jeffreys divergence criterion to study differences in intensity distributions. The effects of these findings are also empirically verified on automatic segmentation of brains with MS.

  13. Effect of Intensive Therapy of Multiple Factors Intervention on Vascular Complications in Type 2 Diabetes

    Institute of Scientific and Technical Information of China (English)

    吴汉妮; 张淑玲; 沈迪

    2003-01-01

    The effects of intensive versus regular therapy on incidence and progress of microalbuminuria in type 2 diabetes were compared. During a follow-up of 3 years, 96 cases of diabetes mellitus were randomized to intensive and regular therapy groups. HbA1c goal was same in the two groups,but the goal of blood pressure (Bp) and lipid was more strict in the intensive therapy group than in the regular therapy group. There was statistically significant difference in the incidence and progression of vascular complications between the two groups. Logistic stepwise-regression analysis (odds ration, OR) showed that there was significant difference in the progression of nephropathy (OR 0. 24,95 % CI 0. 12-0. 76), retinopathy (OR 0.38, 95 % CI 0.16-0. 88), peripheral neuropathy (OR 0. 42, 95 % CI 0. 22-0. 86) and autonomic neuropathy (OR 0. 29, 95 % CI 0. 12-0. 86) between the two groups (P<0. 01). It was concluded that intensive blood glucose controlling could retard diabetic vascular complications. Intensive therapy of multiple factors interventions (controlling Bp, regulating blood lipid, improving microcirculation) could decrease various risk factors for diabetic vascular complications.

  14. Nitrogen fluorescence induced by the femtosecond intense laser pulses in air

    Institute of Scientific and Technical Information of China (English)

    He Li; Suyu Li; Shuchang Li; Dunli Liu; Dan Tian; Anmin Chen; Ying Wang; Xiaowei Wang; Yunfeng Zhang; Mingxing Jin

    2016-01-01

    Our experiments show that external focusing and initial laser energy strongly influences filament generated by the femtosecond Ti–sapphire laser in air. The experimental measurements show the filament length can be extended both by increasing the laser energy and focal length of focusing lens. On the other hand, the plasma fluorescence emission can be enhanced by increasing the laser energy with fixed focal length or decreasing the focal length. In addition, the collapse distance measured experimentally are larger than the calculated ones owing to the group-velocity-dispersion effect. In addition, we find that the line widths of the spectral lines from N2 is independent of filament positions, laser energies and external focusing.

  15. In situ dimerization of multiple wild type and mutant zinc transporters in live cells using bimolecular fluorescence complementation.

    Science.gov (United States)

    Lasry, Inbal; Golan, Yarden; Berman, Bluma; Amram, Noy; Glaser, Fabian; Assaraf, Yehuda G

    2014-03-14

    Zinc transporters (ZnTs) facilitate zinc efflux and zinc compartmentalization, thereby playing a key role in multiple physiological processes and pathological disorders, presumed to be modulated by transporter dimerization. We recently proposed that ZnT2 homodimerization is the underlying basis for the dominant negative effect of a novel heterozygous G87R mutation identified in women producing zinc-deficient milk. To provide direct visual evidence for the in situ dimerization and function of multiple normal and mutant ZnTs, we applied here the bimolecular fluorescence complementation (BiFC) technique, which enables direct visualization of specific protein-protein interactions. BiFC is based upon reconstitution of an intact fluorescent protein including YFP when its two complementary, non-fluorescent N- and C-terminal fragments (termed YN and YC) are brought together by a pair of specifically interacting proteins. Homodimerization of ZnT1, -2, -3, -4, and -7 was revealed by high subcellular fluorescence observed upon co-transfection of non-fluorescent ZnT-YC and ZnT-YN; this homodimer fluorescence localized in the characteristic compartments of each ZnT. The validity of the BiFC assay in ZnT dimerization was further corroborated when high fluorescence was obtained upon co-transfection of ZnT5-YC and ZnT6-YN, which are known to form heterodimers. We further show that BiFC recapitulated the pathogenic role that ZnT mutations play in transient neonatal zinc deficiency. Zinquin, a fluorescent zinc probe applied along with BiFC, revealed the in situ functionality of ZnT dimers. Hence, the current BiFC-Zinquin technique provides the first in situ evidence for the dimerization and function of wild type and mutant ZnTs in live cells.

  16. Using water Raman intensities to determine the effective excitation and emission path lengths of fluorophotometers for correcting fluorescence inner filter effect.

    Science.gov (United States)

    Nettles, Charles B; Hu, Juan; Zhang, Dongmao

    2015-01-01

    Fluorescence and Raman inner filter effects (IFE) cause spectral distortion and nonlinearity between spectral signal intensity with increasing analyte concentration. Convenient and effective correction of fluorescence IFE has been an active research goal for decades. Presented herein is the finding that fluorescence and Raman IFE can be reliably corrected using the equation I(corr)/I(obsd) = 10(dxAx + dmAm) when the effective excitation and emission path lengths, dx and dm, of a fluorophotometer are determined by simple linear curve-fitting of Raman intensities of a series of water Raman reference samples that have known degrees of Raman IFEs. The path lengths derived with one set of Raman measurements at one specific excitation wavelength are effective for correcting fluorescence and Raman IFEs induced by any chromophore or fluorophore, regardless of the excitation and emission wavelengths. The IFE-corrected fluorescence intensities are linearly correlated to fluorophore concentration over 5 orders of magnitude (from 5.9 nM to 0.59 mM) for 2-aminopurine in a 1 cm × 0.17 cm fluorescence cuvette. This water Raman-based method is easy to implement. It does not involve complicated instrument geometry determination or difficult data manipulation. This work should be of broad significance to physical and biological sciences given the popularity of fluorescence techniques in analytical applications.

  17. Computer simulations on resonant fluorescence spectra in atomic gases in two monochromatic laser fields of arbitrary intensity and magnetic field

    Science.gov (United States)

    Karagodova, Tamara Y.

    1996-03-01

    In the intense radiation fields with power density from 104W/cm2 to 109W/cm2 the essential modification of electronic states of atoms occurs displaying, in particular, in modifications of resonant fluorescence (rf) spectra. We use 'Fermi golden rule' for calculations of relative intensities and frequencies for rf multiplet for real multilevel initially unexcited atoms in two monochromatic laser fields of arbitrary intensity resonant to adjacent transitions of (Xi) or (Lambda) types and magnetic field, giving the level splittings of different values from Zeeman to Paschen-Back effect. The dependence of quasienergies on parameters obtained with the help of a sorting program permits us to define the values of parameters for which the states of the system are mixed and so to receive the correct probability amplitudes for instantaneous or adiabatic regimes of switching the perturbation. The analysis of the quasienergies and form of rf spectra permits us to get relations between the form of the spectra and modifications of electronic structure of the atom due to radiation fields and external magnetic field.

  18. Locally adaptive MR intensity models and MRF-based segmentation of multiple sclerosis lesions

    Science.gov (United States)

    Galimzianova, Alfiia; Lesjak, Žiga; Likar, Boštjan; Pernuš, Franjo; Špiclin, Žiga

    2015-03-01

    Neuroimaging biomarkers are an important paraclinical tool used to characterize a number of neurological diseases, however, their extraction requires accurate and reliable segmentation of normal and pathological brain structures. For MR images of healthy brains the intensity models of normal-appearing brain tissue (NABT) in combination with Markov random field (MRF) models are known to give reliable and smooth NABT segmentation. However, the presence of pathology, MR intensity bias and natural tissue-dependent intensity variability altogether represent difficult challenges for a reliable estimation of NABT intensity model based on MR images. In this paper, we propose a novel method for segmentation of normal and pathological structures in brain MR images of multiple sclerosis (MS) patients that is based on locally-adaptive NABT model, a robust method for the estimation of model parameters and a MRF-based segmentation framework. Experiments on multi-sequence brain MR images of 27 MS patients show that, compared to whole-brain model and compared to the widely used Expectation-Maximization Segmentation (EMS) method, the locally-adaptive NABT model increases the accuracy of MS lesion segmentation.

  19. Manipulating fluorescence color and intensity with regular metal nanoparticle-based composite materials

    Energy Technology Data Exchange (ETDEWEB)

    Nikitin, Andrey G., E-mail: nikitin@cinam.univ-mrs.fr [Centre Interdisciplinaire de Nanoscience de Marseille (CINaM, UPR 3118 CNRS), Aix-Marseille University, Campus de Luminy, Case 913, 13288 Marseille, France and Faculty of Physics and Technology, Al-Farabi Kazakh National University, 71 Al-Farabi Ave., 050040 Almaty (Kazakhstan)

    2016-02-01

    This paper first studies the role of structural parameters of ordered metal nanoparticle-based composites in the modification of the spectra and intensity of directional emission from organic molecules. It then investigates the possibilities of white light generation via color conversion using two materials, one emitting in the green and the other one in the red spectral region. The structures under study exhibit enhanced emission within small solid angle in the forward direction due to excitation of the quasiguided modes. These modes modify the angle-dependent local photon density of states and, thus, result in efficient directional outcoupling of radiation.

  20. A Solar-pumped Fluorescence Model for Line-by-line Emission Intensities in the B-X, A-X, and X-X Band Systems of 12C14N

    Science.gov (United States)

    Paganini, L.; Mumma, M. J.

    2016-09-01

    We present a new quantitative model for detailed solar-pumped fluorescent emission of the main isotopologue of CN. The derived fluorescence efficiencies permit estimation and interpretation of ro-vibrational infrared line intensities of CN in exospheres exposed to solar (or stellar) radiation. Our g-factors are applicable to astronomical observations of CN extending from infrared to optical wavelengths, and we compare them with previous calculations in the literature. The new model enables extraction of rotational temperature, column abundance, and production rate from astronomical observations of CN in the inner coma of comets. Our model accounts for excitation and de-excitation of rotational levels in the ground vibrational state by collisions, solar excitation to the {A}2{{{\\Pi }}}{{i}} and {B}2{{{Σ }}}+ electronically excited states followed by cascade to ro-vibrational levels of {X}2{{{Σ }}}+, and direct solar infrared pumping of ro-vibrational levels in the {X}2{{{Σ }}}+ state. The model uses advanced solar spectra acquired at high spectral resolution at the relevant infrared and optical wavelengths and considers the heliocentric radial velocity of the comet (the Swings effect) when assessing the exciting solar flux for a given transition. We present model predictions for the variation of fluorescence rates with rotational temperature and heliocentric radial velocity. Furthermore, we test our fluorescence model by comparing predicted and measured line-by-line intensities for {X}2{{{Σ }}}+ (1-0) in comet C/2014 Q2 (Lovejoy), thereby identifying multiple emission lines observed at IR wavelengths.

  1. The preparation of core-shell magnetic silica nanospheres for enhancing magnetism and fluorescence intensity.

    Science.gov (United States)

    Yoo, Jeong Ha; Kim, Jong Sung

    2013-11-01

    Recently, magnetic and luminescent composite silica with structure of micro- and nanospheres containing both magnetic (Fe3O4) nanoparticles (MPs) and quantum dots (QDs) has attracted great interests. In this study, we have prepared core-shell structure of silica spheres in which magnets are incorporated into silica core and QDs into a mesoporous silica shell by using C18-TMS (octade-cyltrimethoxysilane). MPs were synthesized by a co-precipitation method from ferrous and ferric solutions with a molecular ratio of 2:3. Monodisperse magnetic silica cores have been prepared via sol-gel reaction of TEOS (tetraethoxysilane) and water using base catalyst. The size of magnetic silica nanospheres was confirmed by dynamic laser light scattering system (DLS) and scanning electoron microscope (SEM). The pore volume and surface area were calculated by using BET after calcination. The core-shell structure plays an important role in providing more domains for MPs in silica Core and QDs in silica shell. QDs were incorporated into the mesoporous shell by hydrophobic interactions. Magnetic characterization was performed using a superconducting quantum interference device (SQUID). The optical properties of the particles were characterized with UV/Vis spectrometer, PL spectrometer, and fluorescence microscope.

  2. 时间延迟反馈控制斑图形成%Studying on the Mechanism for the Surface Plasmonics Enhanced the Fluorescence Radiation Intensity

    Institute of Scientific and Technical Information of China (English)

    汪茂胜; 张季谦; 涂玉兵

    2012-01-01

    The technology of fluorescence spectra has important application in the studying about the material structure and the dynamics process of material reciprocity. The bio - sensor or chemical sensor based on the fluorescence has been researched widely. It is a hot topic to improve the sensitivity of fluorescence based sensor. The nano - metal particles or structure can enhance the fluorescence intensity effectively. It is necessary to study the physical mechanism for the surface plasmonic enhanced fluorescence. In this paper, we discuss the principle of fluorescence radiation and the physical mechanism for the surface plasmonic enhanced fluorescence.. There are three theory model including fluorescence resonant energy transfer, surface plasmonics resonant enhanced fluorescence radiation and the Radiating plasmons model.%在Chlorine—iodine—malonic—acid反应扩散体系中,研究了时间延迟反馈对体系时空动力学的控制作用。首先,理论分析发现调节延迟时间和反馈强度会影响体系霍普夫分岔行为。其次,数值模拟发现时间延迟反馈可诱导体系从稳定定态、图灵斑图态向螺旋波态、整体振荡态的转变。

  3. Intensive Farmaco- Surveillance of Interferon Alpha 2b Recombinant in Multiple Sclerosis.

    Directory of Open Access Journals (Sweden)

    Leslie Pérez Ruiz

    2007-12-01

    Full Text Available Background: The interferon alpha 2b recombinant, produced in Cuba, is used in the treatment of different illnesses, such as the multiple sclerosis. For its commercialization it is needed to know its safety scope. Objective: To assess the adverse reactions of interferon alpha 2b recombinant in the treatment of multiple sclerosis. Method: During the period between 1996 and 2006, 70 clinical histories and data collection notebooks of patients included in the randomized double blind national clinical trial phase IV were revised. This trial was developed in the Clinic of Multiple Sclerosis of the Hospital "Dr. Gustavo Aldereguía Lima" in Cienfuegos. With regard to the total of manifested adverse reactions we analyzed type, length, and use of a given treatment to counteract them, intensity level (light, moderate, serious or lethal and causality level (definitive, probable, possible, conditional or not related. Results: 53 patients out of the total presented 207 adverse reactions to interferon. The most frequent were: fever, migraine, chills, arthralgia, asthenia and myalgia, being most of them moderate collateral effects of definitive character. In 197 patients the outcome was favorable. Conclusion: The use of the interferon alpha 2b recombinant was safe in the treatment of the multiple sclerosis in these patients. 

  4. Relating multi-sequence longitudinal intensity profiles and clinical covariates in incident multiple sclerosis lesions.

    Science.gov (United States)

    Sweeney, Elizabeth M; Shinohara, Russell T; Dewey, Blake E; Schindler, Matthew K; Muschelli, John; Reich, Daniel S; Crainiceanu, Ciprian M; Eloyan, Ani

    2016-01-01

    The formation of multiple sclerosis (MS) lesions is a complex process involving inflammation, tissue damage, and tissue repair - all of which are visible on structural magnetic resonance imaging (MRI) and potentially modifiable by pharmacological therapy. In this paper, we introduce two statistical models for relating voxel-level, longitudinal, multi-sequence structural MRI intensities within MS lesions to clinical information and therapeutic interventions: (1) a principal component analysis (PCA) and regression model and (2) function-on-scalar regression models. To do so, we first characterize the post-lesion incidence repair process on longitudinal, multi-sequence structural MRI from 34 MS patients as voxel-level intensity profiles. For the PCA regression model, we perform PCA on the intensity profiles to develop a voxel-level biomarker for identifying slow and persistent, long-term intensity changes within lesion tissue voxels. The proposed biomarker's ability to identify such effects is validated by two experienced clinicians (a neuroradiologist and a neurologist). On a scale of 1 to 4, with 4 being the highest quality, the neuroradiologist gave the score on the first PC a median quality rating of 4 (95% CI: [4,4]), and the neurologist gave the score a median rating of 3 (95% CI: [3,3]). We then relate the biomarker to the clinical information in a mixed model framework. Treatment with disease-modifying therapies (p < 0.01), steroids (p < 0.01), and being closer to the boundary of abnormal signal intensity (p < 0.01) are all associated with return of a voxel to an intensity value closer to that of normal-appearing tissue. The function-on-scalar regression model allows for assessment of the post-incidence time points at which the covariates are associated with the profiles. In the function-on-scalar regression, both age and distance to the boundary were found to have a statistically significant association with the lesion intensities at some time point

  5. Intense frequency upconversion fluorescence of Er3+/Yb3+ co-doped lithium-strontium-lead-bismuth glasses

    Institute of Scientific and Technical Information of China (English)

    Hongtao Sun; Shiqing Xu; Baoyu Chen; Shixun Dai; Shilong Zhao; Lili Hu; Zhonghong Jiang

    2005-01-01

    @@ Infrared-to-visible upconversion fluorescence of Er3+/Yb3+ co-doped lithium-strontium-lead-bismuth (LSPB) glasses for developing potential upconversion lasers has been studied under 975-nm excitation.Based on the results of energy transfer efficiency and upconversion spectra, the optimal Yb3+-Er3+ concentration ratio is found to be 5∶1. Intense green and red emissions centered at 525, 546, and 657 nm,corresponding to the transitions 2H11/2→4I15/2, 4S3/2→4I15/2, and 4F9/2 → 4I15/2, respectively, were observed. The quadratic dependence of the 525-, 546-, and 657-nm emissions on excitation power indicates that a two-photon absorption process occurs under 975-nm excitation. The high-populated 4I11/2 level is supposed to serve as the intermediate state responsible for the upconversion processes. The intense upconversion luminescence of Er3+/Yb3+ co-doped LSPB glasses may be a potentially useful material for developing upconversion optical devices.

  6. Robust detection of multiple sclerosis lesions from intensity-normalized multi-channel MRI

    Science.gov (United States)

    Karpate, Yogesh; Commowick, Olivier; Barillot, Christian

    2015-03-01

    Multiple sclerosis (MS) is a disease with heterogeneous evolution among the patients. Quantitative analysis of longitudinal Magnetic Resonance Images (MRI) provides a spatial analysis of the brain tissues which may lead to the discovery of biomarkers of disease evolution. Better understanding of the disease will lead to a better discovery of pathogenic mechanisms, allowing for patient-adapted therapeutic strategies. To characterize MS lesions, we propose a novel paradigm to detect white matter lesions based on a statistical framework. It aims at studying the benefits of using multi-channel MRI to detect statistically significant differences between each individual MS patient and a database of control subjects. This framework consists in two components. First, intensity standardization is conducted to minimize the inter-subject intensity difference arising from variability of the acquisition process and different scanners. The intensity normalization maps parameters obtained using a robust Gaussian Mixture Model (GMM) estimation not affected by the presence of MS lesions. The second part studies the comparison of multi-channel MRI of MS patients with respect to an atlas built from the control subjects, thereby allowing us to look for differences in normal appearing white matter, in and around the lesions of each patient. Experimental results demonstrate that our technique accurately detects significant differences in lesions consequently improving the results of MS lesion detection.

  7. Clinical effects of intensive insulin therapy treating traumatic shock combined with multiple organ dysfunction syndrome.

    Science.gov (United States)

    Du, Jundong; Liu, Hongming; Liu, Rong; Yao, Yongming; Jiao, Huabo; Zhao, Xiaodong; Yin, Huinan; Li, Zhanliang

    2011-04-01

    The therapeutic effects of intensive insulin therapy in treatment of traumatic shock combined with multiple organ dysfunction syndrome (MODS) were investigated. A total of 114 patients with traumatic shock combined with MODS were randomly divided into two groups: control group (n=56) treated with conventional therapy, and intensive insulin therapy group (n=58) treated with conventional therapy plus continuous insulin pumping to control the blood glucose level at range of 4.4-6.1 mmol/L. White blood cells (WBC) counts, prothrombin time (PT), serum creatinine (SCr), alanine aminotransferase (ALT), serum albumin and PaO(2) were measured before and at the day 1, 3, 5, 7 and 14 after treatment. The incidence of gastrointestinal dysfunction, the incidence of MODS, hospital stay and the mortality were also observed and compared. After intensive insulin therapy, the WBC counts, SCr, ALT and PT were significantly reduced (Pinsulin therapy. The incidence of gastrointestinal dysfunction, the incidence of MODS, the length of hospital stay and the mortality were markedly decreased (Pinsulin therapy is effective for traumatic shock combined with MODS and can decrease the length of hospital stay and the mortality.

  8. Simultaneous determination of multiple (fluoro)quinolone antibiotics in food samples by a one-step fluorescence polarization immunoassay.

    Science.gov (United States)

    Mi, Tiejun; Wang, Zhanhui; Eremin, Sergei A; Shen, Jianzhong; Zhang, Suxia

    2013-10-02

    This paper describes a rapid one-step fluorescence polarization immunoassay (FPIA) for the simultaneous determination of multiple (fluoro)quinolone antibiotics (FQs) in food samples. Several fluorescent tracers were synthesized and evaluated in the FPIA method based on a broad-specificity of monoclonal antibodies toward FQs. The heterogeneous tracer, SAR-5-FAM, was considered as the optimal choice to prepare the immunocomplex single reagent, which allows a rapid and sensitive displacement reaction by addition of analytes. Optimized single-reagent FPIA exhibited broad cross-reactivities in the range of 7.8-172.2% with 16 FQs tested and was capable of determining most FQs at the level of maximum residue limits. Recoveries for spiked milk and chicken muscle samples were from 77.8 to 116%, with relative standard deviation lower than 17.4%. Therefore, this method could be applicable in routine screening analysis of multiple FQ residues in food samples.

  9. Multiple clinical episodes of Plasmodium falciparum malaria in a low transmission intensity setting: exposure versus immunity.

    Science.gov (United States)

    Rono, Josea; Färnert, Anna; Murungi, Linda; Ojal, John; Kamuyu, Gathoni; Guleid, Fatuma; Nyangweso, George; Wambua, Juliana; Kitsao, Barnes; Olotu, Ally; Marsh, Kevin; Osier, Faith Ha

    2015-05-13

    Epidemiological studies indicate that some children experience many more episodes of clinical malaria than their age mates in a given location. Whether this is as a result of the micro-heterogeneity of malaria transmission with some children effectively getting more exposure to infectious mosquitoes than others, or reflects a failure in the acquisition of immunity needs to be elucidated. Here, we investigated the determinants of increased susceptibility to clinical malaria by comparing the intensity of exposure to Plasmodium falciparum and the acquisition of immunity in children at the extreme ends of the over-dispersed distribution of the incidence of clinical malaria. The study was nested within a larger cohort in an area where the intensity of malaria transmission was low. We identified children who over a five-year period experienced 5 to 16 clinical malaria episodes (children at the tail-end of the over-dispersed distribution, n = 35), remained malaria-free (n = 12) or had a single episode (n = 26). We quantified antibodies against seven Plasmodium falciparum merozoite antigens in plasma obtained at six cross-sectional surveys spanning these five years. We analyzed the antibody responses to identify temporal dynamics that associate with disease susceptibility. Children experiencing multiple episodes of malaria were more likely to be parasite positive by microscopy at cross-sectional surveys (X (2) test for trend 14.72 P = 0.001) and had a significantly higher malaria exposure index, than those in the malaria-free or single episode groups (Kruskal-Wallis test P = 0.009). In contrast, the five-year temporal dynamics of anti-merozoite antibodies were similar in the three groups. Importantly in all groups, antibody levels were below the threshold concentrations previously observed to be correlated with protective immunity. We conclude that in the context of a low malaria transmission setting, susceptibility to clinical malaria is not accounted

  10. Using the geometric mean fluorescence intensity index method to measure ZAP-70 expression in patients with chronic lymphocytic leukemia

    Directory of Open Access Journals (Sweden)

    Wu YJ

    2016-02-01

    Full Text Available Yu-Jie Wu, Hui Wang, Jian-Hua Liang, Yi Miao, Lu Liu, Hai-Rong Qiu, Chun Qiao, Rong Wang, Jian-Yong Li Department of Hematology, First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing, People’s Republic of China Abstract: Expression of ζ-chain-associated protein kinase 70 kDa (ZAP-70 in chronic lymphocytic leukemia (CLL is associated with more aggressive disease and can help differentiate CLL from cases expressing mutated or unmutated immunoglobulin heavy chain variable region (IgHV genes. However, standardizing ZAP-70 expression by flow cytometric analysis has proved unsatisfactory. The key point is that ZAP-70 is weakly expressed with a continuous expression pattern rather than a clear discrimination between positive and negative CLL cells, which means that the resulting judgment is subjective. Thus, in this study, we aimed at assessing the reliability and repeatability of ZAP-70 expression using the geometric mean fluorescence intensity (geo MFI index method based on flow cytometry with 256-channel resolution in a series of 402 CLL patients and to compare ZAP-70 with other biological and clinical prognosticators. According to IgHV mutational status, we were able to confirm that the optimal cut-off point for the geo MFI index was 3.5 in the test set. In multivariate analyses that included the major clinical and biological prognostic markers for CLL, the prognostic impact of ZAP-70 expression appeared to have stronger discriminatory power when the geo MFI index method was applied. In addition, we found that ZAP-70-positive patients according to the geo MFI index method had shorter time to first treatment or overall survival (P=0.0002, P=0.0491. This is the first report showing that ZAP-70 expression can be evaluated by a new approach, the geo MFI index, which could be a useful prognostic method as it is more reliable, less subjective, and therefore better associated with improvement of CLL prognostication

  11. Binomial distribution-based quantitative measurement of multiple-acceptors fluorescence resonance energy transfer by partially photobleaching acceptor

    Science.gov (United States)

    Zhang, Lili; Yu, Huaina; Zhang, Jianwei; Chen, Tongsheng

    2014-06-01

    We report that binomial distribution depending on acceptor photobleaching degree can be used to characterize the proportions of various kinds of FRET (Fluorescence Resonance Energy Transfer) constructs resulted from partial acceptor photobleaching of multiple-acceptors FRET system. On this basis, we set up a rigorous quantitation theory for multiple-acceptors FRET construct named as Mb-PbFRET which is not affected by the imaging conditions and fluorophore properties. We experimentally validate Mb-PbFRET with FRET constructs consisted of one donor and two or three acceptors inside living cells on confocal and wide-field microscopes.

  12. Evaluation of Stress Intensity Factors for Multiple Cracked Circular Disks Under Crack Surface Tractions with SBFEM

    Institute of Scientific and Technical Information of China (English)

    LIU Jun-yu; LIN Gao; LI Xiao-chuan; XU Feng-lin

    2013-01-01

    Stress intensity factors (SIFs) for the cracked circular disks under different distributing surface tractions are evaluated with the scaled boundary finite element method (SBFEM).In the SBFEM,the analytical advantage of the solution in the radial direction allows SIFs to be directly determined from its definition,therefore no special crack-tip treatment is necessary.Furthermore anisotropic material behavior can be treated easily.Different distributions of surface tractions are considered for the center and double-edge-cracked disks.The benchmark examples are modeled and an excellent agreement between the results in the present study and those in published literature is found.It shows that SBFEM is effective and possesses high accuracy.The SIFs of the cracked orthotropic material circular disks subjected to different surface tractions are also evaluated.The technique of substructure is applied to handle the multiple cracks problem.

  13. Optimized fluorescence diagnosis of tumors by comparing five-ALA-induced xenofluorescence and autofluorescence intensities of a murine tumor/nontumor tissue system cultivated on the CAM

    Science.gov (United States)

    Stroebele, Simone; Dressler, Cathrin; Ismail, M. Samy; Daskalaki, Anita; Philipp, Carsten M.; Berlien, Hans-Peter; Weitzel, H.; Liebsch, M.; Spielmann, H.

    1995-12-01

    The in vivo model of the chorioallantoic membrane of fertilized chicken embryos (CAM) was employed for studying the fluorescence characteristics of tumor tissue in comparison with non tumorous tissue. Tumors were grown from the murine fibrosarcoma cell line SSK II and murine 3T3 fibroblasts (clone A31) were used for cultivating non tumorous tissue. Autofluorescence and xenofluorescence intensities induced by 5-aminolaevulinic acid (5-ALA) were compared. Exogenous administration of 5-ALA, an early precursor in haem synthesis, induces accumulation of endogenous photoactive porphyrins, in particular protoporphyrin IX (PpIX). Fluorescence investigations were performed after 3-4d of incubation, when the tissues had reached macroscopically three dimensional stages of growth. Fluorescences were excited with a HBO-X 100 W lamp (Carl Zeiss) at a wavelength (lambda) equals 405 plus or minus 5 nm. Emissions were detected in the spectral range above 630 nm and visualized by real time digital image processing (Argus 10, HAMAMATSU) using an ICCD camera (HAMAMATSU). After administration of 0.4 mmolar 5-ALA solution to the CAM inoculated tissues the SSK II tumors exhibited higher fluorescence intensities than the 3T3 non tumorous tissues. Autofluorescence intensities of both types of tissues were not distinguishable. Furthermore, the effects of several biochemicals on the xenofluorescence intensities of the fibrosarcoma and fibroblast tissues were investigated.

  14. An expression for the atomic fluorescence and thermal-emission intensity under conditions of near saturation and arbitrary self-absorption

    NARCIS (Netherlands)

    Omenetto, N.; Winefordner, J.D.; Alkemade, C.T.J.

    1975-01-01

    An expression for the effect of self-absorption on the fluorescence and thermal emission intensities is derived by taking into account stimulated emission. A simple, idealized case is considered, consisting of a two level atomic system, in a flame, homogeneous with respect to temperature and composi

  15. Formaldehyde Emission in Comet C/2002 T7 (LINEAR) at Infrared Wavelengths: Line-by-Line Validation of Modeled Fluorescent Intensities

    Science.gov (United States)

    DiSanti, Michael A.; Bonev, B. P.; Dello Russo, N.; Magee-Sauer, K.; Mumma, M. J.; Reuter, D. C.; Villanueva, G. L.; Anderson, W. M.; Gibb, E. L.

    2006-09-01

    Cometary nuclei are the most primitive remnants of the early Solar System, so measuring abundances of their ices allows a glimpse into the conditions in which icy bodies formed. Only in the last several years has it become possible to routinely study native cometary volatiles at infrared wavelengths. In spring 2004, we observed comet C/2002 T7 (LINEAR, hereafter C/T7) using the CSHELL spectrometer at the NASA-IRTF 3-m telescope. CSHELL offers sufficiently high spectral resolving power (λ/δλ 2.5 x 104) to permit line-by-line intensities to be measured. Emission lines from multiple molecular species were targeted in the 2.9 - 5.0 micron spectral region, and our observations revealed an extremely rich chemistry in C/T7, including formaldehyde (H2CO). H2CO is ubiquitous in dense interstellar clouds, so its presence is expected in comets if they contain interstellar material. These bodies delivered enormous quantities of pre-biotic chemicals to the young Earth, thus the abundance of H2CO in comets is of keen astrobiological interest. C/T7 provided the best opportunity to date to compare H2CO line intensities predicted by an existing fluorescence model1 with high-resolution comet spectra. We present results2 showing a high degree of correlation between model and data. This work validates the model and permits highly reliable measures of rotational temperature and production rate of H2CO, for comparison with chemically related molecules (CO, methyl alcohol) in C/T7 and other comets in our database. This research was supported by the NASA Planetary Atmospheres (RTOP 344-33-55), Astrobiology (RTOP 344-53-51), and Planetary Astronomy (RTOP 344-32-98) Programs. References: 1Reuter, D. C., et al. 1989 Ap J 341:1045-1058; 2DiSanti, M. A., et al. 2006 Ap J (in press)

  16. Intense-Field Multiple-Detachment of F2¯: Competition with Photodissociation.

    Science.gov (United States)

    Shahi, Abhishek; Albeck, Yishai; Strasser, Daniel

    2017-04-07

    The competition of intense-field multiple-detachment with efficient photodissociation of F2¯ is studied as a function of laser peak intensity. The main product channels are disentangled and characterized by 3D coincidence fragment imaging. The presented kinetic energy release spectra, angular distributions as well as two color pump-probe measurements allow identification of competing sequential and non-sequential mechanisms. Dissociative detachment, producing two neutral atoms (F + F) is found to be dominated by a sequential mechanism of photodissociation (F¯ + F) followed by detachment of the atomic anion fragment. In contrast, dissociative ionization (F + F(+)) shows competing contributions of both a sequential two-step mechanism as well as a non-sequential double-detachment of the molecular anion, which are distinguished by the kinetic energy released in the dissociation. Triple-detachment is found to be non-sequential in nature and results in Coulomb explosion (F(+)+F(+)). Furthermore, the measured kinetic energy release for dissociation on the (2)Σg(+) state provides a direct measurement of the F2¯ dissociation energy, D0 = 1.26±0.03 eV.

  17. The impact of relative intensity noise on the signal in multiple reference optical coherence tomography

    Science.gov (United States)

    Neuhaus, Kai; Subhash, Hrebesh; Alexandrov, Sergey; Dsouza, Roshan; Hogan, Josh; Wilson, Carol; Leahy, Martin; Slepneva, Svetlana; Huyet, Guillaume

    2016-03-01

    Multiple reference optical coherence tomography (MR-OCT) applies a unique low-cost solution to enhance the scanning depth of standard time domain OCT by inserting an partial mirror into the reference arm of the interferometric system. This novel approach achieves multiple reflections for different layers and depths of an sample with minimal effort of engineering and provides an excellent platform for low-cost OCT systems based on well understood production methods for micro-mechanical systems such as CD/DVD pick-up systems. The direct integration of a superluminescent light-emitting diode (SLED) is a preferable solution to reduce the form- factor of an MR-OCT system. Such direct integration exposes the light source to environmental conditions that can increase fluctuations in heat dissipation and vibrations and affect the noise characteristics of the output spectrum. This work describes the impact of relative intensity noise (RIN) on the quality of the interference signal of MR-OCT related to a variety of environmental conditions, such as temperature.

  18. Aggregation-induced fluorescence behavior of triphenylamine-based Schiff bases: the combined effect of multiple forces.

    Science.gov (United States)

    Yang, Mingdi; Xu, Dongling; Xi, Wengang; Wang, Lianke; Zheng, Jun; Huang, Jing; Zhang, Jingyan; Zhou, Hongping; Wu, Jieying; Tian, Yupeng

    2013-10-18

    Eight triphenylamine (TPA)-based Schiff bases that exhibit different aggregation-induced emission (AIE) or aggregation-caused quenching (ACQ) behavior in tetrahydrofuran (THF)/water mixtures have been synthesized and characterized. The photophysical properties in solution, aqueous suspension, film, and the crystalline state along with their relationships were comparatively investigated. The single-crystal structures of 1-8 indicate that compact π···π stacking or excimers induce fluorescence quenching of 1, 2, 5, and 7. However, the existence of J aggregates or multiple intra- and intermolecular interactions restrict the intramolecular vibration and rotation, enabling compounds 3, 4, 6, and 8 to exhibit good AIE character. The size and growth process of particles with different water fractions were studied using scanning electron microscopy, which demonstrated that smaller uniformly dispersed nanoparticles in the THF/water mixtures favor fluorescence emission. The above results suggest that the combined effects of multiple forces caused by structural variation have a great influence on their molecular packing, electronic structure, and aggregation-induced fluorescence properties. In addition, piezofluorochromic experiments verified the potential applications of 4 and 6.

  19. A dithienosilole-based fluorescent chemosensor for multiple logic operations at the molecular level.

    Science.gov (United States)

    Zhang, Chen; Sun, Caixia; Lu, Yahong; Wang, Junxing; He, Xingxing; Lu, Junting; Yin, Shouchun; Qiu, Huayu

    2015-11-01

    A chemosensor consisting of two terpyridines covalently linked to a dithienosilole unit (1) has been synthesized, and its optical and metal sensing properties have been investigated. Due to the metal-organic coordination function, 1 can bind with many transition metal ions and display different fluorescence responses that cause it to function as a "turn-off" fluorescent chemosensor. A significant bathochromic shift in the fluorescence spectra is observed in the presence of Zn(2+). Meanwhile, the emission of 1 is weakened upon exposure to Ag(+) and Fe(2+) and completely quenched by Ni(2+), Co(2+), and Cu(2+). Based on the observed results, several logic gates, such as XNOR, INHIBIT, and IMPLICATION, have been achieved by controlling the chemical inputs.

  20. Time course of strength adaptations following high-intensity resistance training in individuals with multiple sclerosis.

    Science.gov (United States)

    Manca, A; Dvir, Z; Dragone, D; Mureddu, G; Bua, G; Deriu, Franca

    2017-04-01

    No evidence exists regarding the time course and clinical relevance of muscle strength improvements following resistance training in people with multiple sclerosis (PwMS). The purpose of this study was to investigate the temporal course and the clinical meaningfulness of the changes in strength induced by high-intensity resistance training and whether these changes impact on muscle endurance to fatigue and functional outcomes. PwMS with predominantly unilateral hyposthenia of the ankle dorsiflexors underwent a 6-week isokinetic training of the more affected ankle dorsiflexion muscles. Maximal strength was measured at baseline, during the training on a weekly basis, at the end of the intervention (POST) and at the 12-week follow-up. Muscle endurance to fatigue, mobility and walking outcomes were assessed at baseline, POST and follow-up. Reproducibility and responsiveness analyses were performed. Significant gains in muscle strength were already detected after 3 weeks of training with no further improvements in the following weeks. These improvements exceeded the cutoff values for relevant changes and were also positively correlated to improved muscle endurance to fatigue and mobility measures. None of the observed changes in muscle performance and functional outcomes was retained at the follow-up. Preliminary evidence showed that 3 weeks of high-intensity resistance training induces consistent and meaningful improvements in muscle performance of the ankle dorsiflexors in PwMS. These findings may have practical dose-response and cost-effectiveness implications in the management of MS-induced muscle weakness, potentially enhancing the understanding of the response to training exhibited by PwMS. ClinicalTrials.gov identifier NCT02010398; December 2013.

  1. Multiple neuroanatomical tract-tracing using fluorescent Alexa Fluor conjugates of cholera toxin subunit B in rats.

    Science.gov (United States)

    Conte, William L; Kamishina, Hiroaki; Reep, Roger L

    2009-01-01

    Cholera toxin subunit B (CTB) is a highly sensitive retrograde neuroanatomical tracer. With the new availability of fluorescent Alexa Fluor (AF) conjugates of CTB, multiple neuroanatomical connections can be reliably studied and compared in the same animal. Here we provide a protocol that describes the use of AF-CTB for studying connections in the central nervous system of rats. The viscous properties of CTB allow small and discreet injection sites yet still show robust retrograde labeling. Furthermore, the AF conjugates are extremely bright and photostable, compared with other conventional fluorescent tracers. This protocol can also be adapted for use with other neuroanatomical tracers. Including a 7-d survival period, this protocol takes approximately 11 to 12 d to complete in its entirety.

  2. Fluorescence lifetime spectroscopy in multiple-scattering environments: an application to biotechnology

    Science.gov (United States)

    Cerussi, Albert E.; Gratton, Enrico; Fantini, Sergio

    1999-07-01

    Over the past few years, there has been significant research activity devoted to the application of fluorescence spectroscopy to strongly scattering media, where photons propagate diffusely. Much of this activity focused on fluorescence as a source of contrast enhancement in optical tomography. Our efforts have emphasized the quantitative recovery of fluorescence parameters for spectroscopy. Using a frequency-domain diffusion-based model, we have successfully recovered the lifetime, the absolute quantum yield, the fluorophore concentration, and the emission spectrum of the fluorophore, as well as the absorption and the reduced scattering coefficients at the emission wavelength of the medium in different measurements. In this contribution, we present a sensitive monitor of the binding between ethidium bromide and bovine cells in fresh milk. The spectroscopic contrast was the approximately tenfold increase in the ethidium bromide lifetime upon binding to DNA. The measurement clearly demonstrated that we could quantitatively measure the density of cells in the milk, which is an application vital to the tremendous economic burden of bovine subclinical mastitis detection. Furthermore, we may in principle use the spirit of this technique as a quantitative monitor of the binding of fluorescent drugs inside tissues. This is a first step towards lifetime spectroscopy in tissues.

  3. Intravital Fluorescence Facilitates Measurement of Multiple Physiologic Functions and Gene Expression in Tumors of Live Animals

    Directory of Open Access Journals (Sweden)

    Mark W. Dewhirst

    2002-01-01

    Full Text Available The purpose of this report is to present an overview of the use of fluorescence imaging in vivo, with particular emphasis on oncology. It is important to note, however, that many of the methods described herein have been applied to the study of non-malignant tissues as well. Modern medicine and biology research has benefited greatly from an ever-expanding assortment of fluorescent markers and labels. These markers and labels have allowed investigators to observe the behavior and properties of cell and molecular entities of interest in the context of complicated biological systems such as a mammalian cell or a whole mouse. Methods developed to image fluorescence in whole mice have been valuable in studying patterns of tumor growth and metastases. Alternatively, more detailed information and a wide variety of endpoints can be obtained using “intravital” preparations. This review focuses on use of fluorescence imaging for intravital preparations. For detail on fluorescence imaging of whole animals, refer to reviews on this subject [1,2]. For oncologic applications, studies have focused primarily on window chamber preparations that allow for real-time visualization of tumor growth, vascularity, vascular responses to stimulation, vascular permeability, vascular orientation, flow instability, and the like. These endpoints have been used to show that there are functional differences between tumor and normal tissues with respect to these functions under baseline conditions and after therapeutic manipulation. Examples of some of these differences are provided in this review as a means to illustrate how they can be used.

  4. Multiple time scales in modeling the incidence of infections acquired in intensive care units

    Directory of Open Access Journals (Sweden)

    Martin Wolkewitz

    2016-09-01

    Full Text Available Abstract Background When patients are admitted to an intensive care unit (ICU their risk of getting an infection will be highly depend on the length of stay at-risk in the ICU. In addition, risk of infection is likely to vary over calendar time as a result of fluctuations in the prevalence of the pathogen on the ward. Hence risk of infection is expected to depend on two time scales (time in ICU and calendar time as well as competing events (discharge or death and their spatial location. The purpose of this paper is to develop and apply appropriate statistical models for the risk of ICU-acquired infection accounting for multiple time scales, competing risks and the spatial clustering of the data. Methods A multi-center data base from a Spanish surveillance network was used to study the occurrence of an infection due to Methicillin-resistant Staphylococcus aureus (MRSA. The analysis included 84,843 patient admissions between January 2006 and December 2011 from 81 ICUs. Stratified Cox models were used to study multiple time scales while accounting for spatial clustering of the data (patients within ICUs and for death or discharge as competing events for MRSA infection. Results Both time scales, time in ICU and calendar time, are highly associated with the MRSA hazard rate and cumulative risk. When using only one basic time scale, the interpretation and magnitude of several patient-individual risk factors differed. Risk factors concerning the severity of illness were more pronounced when using only calendar time. These differences disappeared when using both time scales simultaneously. Conclusions The time-dependent dynamics of infections is complex and should be studied with models allowing for multiple time scales. For patient individual risk-factors we recommend stratified Cox regression models for competing events with ICU time as the basic time scale and calendar time as a covariate. The inclusion of calendar time and stratification by ICU

  5. Determination of multiple phytohormones in fruits by high-performance liquid chromatography with fluorescence detection using dispersive liquid-liquid microextraction followed by precolumn fluorescent labeling.

    Science.gov (United States)

    Li, Guoliang; Lu, Shuaimin; Wu, Hongliang; Chen, Guang; Liu, Shucheng; Kong, Xiaojian; Kong, Weiheng; You, Jinmao

    2015-01-01

    Plant hormone determination in food matrices has attracted more and more attention because of their potential risks to human health. However, analytical methods for the analysis of multiple plant hormones remain poorly investigated. In the present study, a convenient, selective, and ultrasensitive high-performance liquid chromatography method for the simultaneous determination of multiple classes of plant hormones has been developed successfully using dispersive liquid-liquid microextraction followed by precolumn fluorescent labeling. Eight plant hormones in fruits including jasmonic acid, 12-oxo-phytodienoic acid, indole-3-acetic acid, 3-indolybutyric acid, 3-indolepropionic acid, gibberellin A3 , 1-naphthylacetic acid, and 2-naphthaleneacetic acid were analyzed by this method. The conditions employed for dispersive liquid-liquid microextraction were optimized systematically. The linearity for all plant hormones was found to be >0.9993 (R(2) values). This method offered low detection limits of 0.19-0.44 ng/mL (at a signal-to-noise ratio of 3), and method accuracies were in the range of 92.32-103.10%. The proposed method was applied to determine plant hormones in five kinds of food samples, and this method can achieve a short analysis time, low threshold levels of detection, and a high specificity for the analysis of targeted plant hormones present at trace level concentrations in complex matrices.

  6. Multiple organ dysfunction syndrome associated with hyperglycemia in children requiring intensive care

    Directory of Open Access Journals (Sweden)

    Hendy Halim

    2015-07-01

    Conclusion Multiple organ dysfunction syndrome had a significant association with hyperglycemia. Multiple organ dysfunction syndrome with hyperglycemia occurs ten times greater than with normoglycemia.

  7. Multiple temperature effects on up-conversion fluorescences of Er3+-Y b3+-Mo6+ codoped TiO2 and high thermal sensitivity

    Directory of Open Access Journals (Sweden)

    B. S. Cao

    2015-08-01

    Full Text Available We report multiple temperature effects on green and red up-conversion emissions in Er3+-Y b3+-Mo6+ codoped TiO2 phosphors. With increasing temperature, the decrease of the red emission from 4F9/2→4I15/2, the increase of green emission from 2H11/2→4I15/2 and another unchanged green emission from 4S3/2→4I15/2 were simultaneously observed, which are explained by steady-state rate equations analysis. Due to different evolution with temperature of the two green emissions, higher thermal sensitivity of optical thermal sensor was obtained based on the transitions with the largest fluorescence intensity ratio. Two parameters, maximum theoretical sensitivity (Smax and optimum operating temperature (Tmax are given to describe thermal sensing properties of the produced sensors. The intensity ratio and energy difference ΔE of a pair of energy levels are two main factors for the sensitivity and accuracy of sensors, which should be referred to design sensors with optimized sensing properties.

  8. Validation of interphase fluorescence in situ hybridization (iFISH for multiple myeloma using CD138 positive cells

    Directory of Open Access Journals (Sweden)

    Renata Kiyomi Kishimoto

    2016-06-01

    Full Text Available ABSTRACT BACKGROUND: Multiple myeloma is a plasma cell neoplasm with acquired genetic abnormalities of clinical and prognostic importance. Multiple myeloma differs from other hematologic malignancies due to a high fraction of low proliferating malignant plasma cells and the paucity of plasma cells in bone marrow aspiration samples, making cytogenetic analysis a challenge. An abnormal karyotype is found in only one-third of patients with multiple myeloma and interphase fluorescence in situ hybridization is the most useful test for studying the chromosomal abnormalities present in almost 90% of cases. However, it is necessary to study the genetic abnormalities in plasma cells after their identification or selection by morphology, immunophenotyping or sorting. Other challenges are the selection of the most informative FISH panel and determining cut-off levels for FISH probes. This study reports the validation of interphase fluorescence in situ hybridization using CD138 positive cells, according to proposed guidelines published by the European Myeloma Network (EMN in 2012. METHOD: Bone marrow samples from patients with multiple myeloma were used to standardize a panel of five probes [1q amplification, 13q14 deletion, 17p deletion, t(4;14, and t(14;16] in CD138+ cells purified by magnetic cell sorting. RESULTS: This test was validated with a low turnaround time and good reproducibility. Five of six samples showed genetic abnormalities. Monosomy/deletion 13 plus t(4;14 were found in two cases. CONCLUSION: This technique together with magnetic cell sorting is effective and can be used in the routine laboratory practice. In addition, magnetic cell sorting provides a pure plasma cell population that allows other molecular and genomic studies.

  9. Isolation and selection of fluorescent pseudomonads based on multiple plant growth promotion traits and siderotyping

    Directory of Open Access Journals (Sweden)

    Jayamohan Subramanian

    2014-09-01

    Full Text Available Fluorescent pseudomonads, acclaimed plant associated bacterial group, are well-known plant growth promoting-biocontrol agents in rhizosphere arena. In this study, 144 fluorescent pseudomonad isolates from rhizosphere soil samples were screened with King's medium B supplemented with 8-hydroxyquinoline (8-HQ chelator and comprehensively profiled for plant growth promotion viz., production of indole acetic acid (IAA, siderophore, ammonia, hydrogen cyanide, motility, phosphate solubilization, root growth promotion, and biofilm forming ability, along with two known control strains of pseudomonads. Iron and IAA regulated secondary metabolite siderophore production were investigated quantitatively. All isolates were positive for ammonia production and motility; 46% isolates were positive for hydrogen cyanide, 44% shown positivity for phosphate solubilization, and 40% isolates for siderophore production. Siderotyping showed production of hydroxamate type of siderophores which are known to be more efficient biocontrol agents. All isolates stimulated root growth to varying extent and had potentiality to form biofilms, a critical constituent for survival on different environments. Forty-two isolates of pseudomonads showed antagonistic behavior against the deleterious fungal pathogen Fusarium oxysporum (MTCC1755. Based on the above observations and statistical analysis, 11 isolates were shortlisted for further scrutiny. The study of biogeographic correlation and secondary metabolite profiling in association with plant growth promotion focalizes significant assessment on the behavior and antagonistic action, which probably brings out a competent biocontrol agent in a sustainable eco-friendly dimension.

  10. [Research on the Relationship between Surface Structure and Fluorescence Intensity of Ca(1-x)Al2Si2O8 : Eu(x)].

    Science.gov (United States)

    He, Xiao; Zhang, Li-sheng; Zu, En-dong; Yang, Xiao-yun; Dong, Kun

    2016-01-01

    Ca(1-x)Al2Si2O8 : Eu(x)(x = 0, 0.01, 0.05, 0.15) were synthesized by solid-state reaction respectively at 1 150, 1 250 1350 and 1 450 degrees C. With X-ray diffraction(XRD), Raman spectroscopy(Raman), photoluminescence spectroscopy(PL) and X-ray fluorescence spectrometer(XRF), the relationship between surface structure and fluorescence intensity of Ca(1-x) Al2Si2O8: Eu(x) were studied. XRD and Raman results show that, CaAl2Si2O8 anorthite single-phase has formed gradually along with the temperature rising in the process of synthesis. Raman spectroscopy is clear that when the Eu doping amount is the same, Si-O amorphous phase disappear gradually and the CaAl2Si2O8 phase form gradually with the temperature increases. As the temperature increases, vibration peaks position silicon oxygen tetrahedron shift to lower wave number. When 1 450 degrees C, the temperature is too high to destroy the structure of silicon oxygen tetrahedron. At the same time, there is a broadening amorphous peak appears in Raman spectroscopy. The procedure of Al to replace Si is hindered with Eu doped in. It is the result that the peak at 1 620 cm(-1) decreases after the first increases. The change of surface structure associated with the scattering amount of Eu. PL and XRF results show that: as the temperature increases, the amount of Eu atom scattering on the material surface increases gradually, this change lead to the fluorescence intensity raise. Therefore, there is proportional relationship between the fluorescence intensity of the samples and the number of samples per unit surface area of Eu atoms.

  11. Simultaneous tracking of movement and gene expression in multiple Drosophila melanogaster flies using GFP and DsRED fluorescent reporter transgenes

    Directory of Open Access Journals (Sweden)

    Tavaré Simon

    2009-04-01

    Full Text Available Abstract Background Fluorescent proteins such as GFP (Green Fluorescent Protein and DsRED (Discosoma sp.Red Fluorescent Protein are often used as reporter molecules for transgene expression in Drosophila and other species. We have recently reported methods that allow simultaneous tracking of animal movement and GFP expression in real time, however the assay was limited to single animals and a single transgene. Numerous studies would be facilitated by methods that allow for assay of multiple animals and multiple transgenes. Findings Here we report an improved fly video tracking system that allows multiple transgenic flies to be tracked simultaneously using visible light, GFP fluorescence and DsRED fluorescence. The movement of multiple flies could be accurately tracked at real time rates, while simultaneously assaying the expression level of two different transgenes marked with GFP and DsRED. The individual flies could be accurately tracked and distinguished even during periods when transgene fluorescence was undetected. For example, characteristic patterns of hsp70 and hsp22 transgene induction could be simultaneously quantified and correlated with animal movement in aging flies, and as groups of flies died due to dessication/starvation. Conclusion The improved methods allow for more efficient assay of the correlation between gene expression, behavior, aging and mortality: multiple animals can be assayed with simultaneous quantification of multiple transgenes using GFP and DsRED fluorescence. These methods should allow for increased flexibility in experimental designs. For example, in the future it should be possible to use gene expression levels to predict remaining life span more accurately, and to quantify gene expression changes caused by interactions between animals in real time.

  12. Absence of multiple local minima effects in intensity modulated optimization with dose-volume constraints

    Energy Technology Data Exchange (ETDEWEB)

    Llacer, Jorge [EC Engineering Consultants, LLC 130, Forest Hill Drive, Los Gatos, CA (United States); Deasy, Joseph O [Department of Radiation Oncology, Mallinckrodt Institute of Radiology, Washington University School of Medicine, St. Louis, MO (United States); Bortfeld, Thomas R [Department of Radiation Oncology, Massachusetts General Hospital and Harvard Medical School, 30 Fruit Street, Boston, MA (United States); Solberg, Timothy D [Department of Radiation Oncology, University of California, Los Angeles, CA (United States); Promberger, Claus [BrainLAB AG, Ammerthalstrasse 8, 85551 Heimstetten (Germany)

    2003-01-21

    This paper reports on the analysis of intensity modulated radiation treatment optimization problems in the presence of non-convex feasible parameter spaces caused by the specification of dose-volume constraints for the organs-at-risk (OARs). The main aim was to determine whether the presence of those non-convex spaces affects the optimization of clinical cases in any significant way. This was done in two phases: (1) Using a carefully designed two-dimensional mathematical phantom that exhibits two controllable minima and with randomly initialized beamlet weights, we developed a methodology for exploring the nature of the convergence characteristics of quadratic cost function optimizations (deterministic or stochastic). The methodology is based on observing the statistical behaviour of the residual cost at the end of optimizations in which the stopping criterion is progressively more demanding and carrying out those optimizations to very small error changes per iteration. (2) Seven clinical cases were then analysed with dose-volume constraints that are stronger than originally used in the clinic. The clinical cases are two prostate cases differently posed, a meningioma case, two head-and-neck cases, a spleen case and a spine case. Of the 14 different sets of optimizations (with and without the specification of maximum doses allowed for the OARs), 12 fail to show any effect due to the existence of non-convex feasible spaces. The remaining two sets of optimizations show evidence of multiple minima in the solutions, but those minima are very close to each other in cost and the resulting treatment plans are practically identical, as measured by the quality of the dose-volume histograms (DVHs). We discuss the differences between fluence maps resulting from those similar treatment plans. We provide a possible reason for the observed results and conclude that, although the study is necessarily limited, the annealing characteristics of a simulated annealing method may not be

  13. Absence of multiple local minima effects in intensity modulated optimization with dose-volume constraints

    Science.gov (United States)

    Llacer, Jorge; Deasy, Joseph O.; Bortfeld, Thomas R.; Solberg, Timothy D.; Promberger, Claus

    2003-01-01

    This paper reports on the analysis of intensity modulated radiation treatment optimization problems in the presence of non-convex feasible parameter spaces caused by the specification of dose-volume constraints for the organs-at-risk (OARs). The main aim was to determine whether the presence of those non-convex spaces affects the optimization of clinical cases in any significant way. This was done in two phases: (1) Using a carefully designed two-dimensional mathematical phantom that exhibits two controllable minima and with randomly initialized beamlet weights, we developed a methodology for exploring the nature of the convergence characteristics of quadratic cost function optimizations (deterministic or stochastic). The methodology is based on observing the statistical behaviour of the residual cost at the end of optimizations in which the stopping criterion is progressively more demanding and carrying out those optimizations to very small error changes per iteration. (2) Seven clinical cases were then analysed with dose-volume constraints that are stronger than originally used in the clinic. The clinical cases are two prostate cases differently posed, a meningioma case, two head-and-neck cases, a spleen case and a spine case. Of the 14 different sets of optimizations (with and without the specification of maximum doses allowed for the OARs), 12 fail to show any effect due to the existence of non-convex feasible spaces. The remaining two sets of optimizations show evidence of multiple minima in the solutions, but those minima are very close to each other in cost and the resulting treatment plans are practically identical, as measured by the quality of the dose-volume histograms (DVHs). We discuss the differences between fluence maps resulting from those similar treatment plans. We provide a possible reason for the observed results and conclude that, although the study is necessarily limited, the annealing characteristics of a simulated annealing method may not be

  14. The sensitive capillary electrophoretic-LIF method for simultaneous determination of curcuminoids in turmeric by enhancing fluorescence intensities of molecules upon inclusion into (2-hydroxypropyl)-β-cyclodextrin.

    Science.gov (United States)

    Kalaycıoğlu, Zeynep; Hashemi, Parya; Günaydın, Keriman; Erim, F Bedia

    2015-10-01

    Curcuminoids have received great attention in the past decades due to their health benefit properties. The aim of this study is to develop a very simple, rapid, and sensitive capillary zone electrophoresis technique coupled with a laser induced fluorescence detector (LIF) for the simultaneous determination of three major curcuminoids of turmeric, namely, curcumin, demethoxy curcumin (DMC), and bisdemethoxy curcumin (BDMC). Background electrolyte was selected as borate at pH 9.6 and (2-hydroxypropyl)-β-cyclodextrin (2-HP-β-CD) was added to prevent rapid alkali degradation of curcuminoids in buffer and to increase fluorescence intensities of molecules. With the addition of 2-HP-β-CD to the separation electrolyte, the fluorescence signal intensities of curcuminoids were enhanced considerably by 30, 40, and 54 fold for curcumin, DMC, and BDMC, respectively. The three curcuminoids of turmeric were fully separated and quantified in less than 4.5 min. The repeatability of the peak areas of curcuminoids for intra-day and inter-day experiments was in the satisfactory range of 2.26 and 2.55%, respectively. The LOD and LOQ values for the developed method were equal to or less than 0.081 and 0.270 μg/mL, respectively, for all curcuminoids. The developed method was successfully applied to find curcuminoids amount in turmeric samples and herbal supplements.

  15. Measurement of K-X-rays fluorescence cross-sections, fluorescence yields and intensity ratios for elements in the atomic range 21 < Z < 74 excited by 59 keV photons

    Energy Technology Data Exchange (ETDEWEB)

    Gomez-Avila, J.; Lopez-Pino, N.; Padilla-Cabal, F.; Van Espen, P.; Cabal, A.; Pena, M. Ruiz; Alessandro, K.D.; Maidana, N.L. [Instituto Superior de Tecnologia y Ciencias Aplicadas (InSTEC), La Habana (Cuba); Antwerp Univ. (Belgium). Micro Trace Analytical Center; CEADEN, La Habana (Cuba); Universidade de Sao Paulo (IF/USP), SP (Brazil). Inst. de Fisica. Lab. do Acelerador Linear

    2010-07-01

    Full text: Using 59 keV photons, we measured the K{sub {alpha}}, K{sub {beta}} and total K X-rays fluorescence cross sections of 17 elements in the atomic range 21 < Z < 74. Furthermore, the fluorescence yields and the I{sub K{beta}} / I{sub K{alpha}} intensity ratios for these elements were also determined. An annular radioactive source of {sup 241}Am (activity 1 Ci) was employed to excite the elements in targets with the shape of foils or pellets (99% purity and 20 mm, in diameter). The pellets were formed with a mixture of cellulose and a chemical compound containing the element of interest, pressed at 15 tons. The K X-rays emitted from the irradiated samples were detected by a Si(Li) detector with a frontal Pb collimator, coupled to conventional electronics, with dead time below 10%. The fluxes reaching the targets and the crystal detector were determined by means of Monte Carlo (MC) simulations using the MCNPX V 2.6 code. The input geometries included the detector, the sample-source holder and the Pb collimator. The optimal diameter for the samples as well as the collimator dimensions were estimated by means of MC simulations. Using several elements (Ti, Ni, Br, Ag, Cs, Dy and W) a calibration curve for the effective flux of photons (I{sub 0}G{sub {epsilon}}) as function of the K X-rays energy was measured. Correction by different sizes and self-absorption coefficients of the samples were also performed. The data obtained for the X-rays fluorescence cross sections were compared with semi-empirical calculations and with experimental values reported by other authors; the relative deviations were less than 10%. Keywords: fluorescence cross section, fluorescence yields, Monte Carlo (author)

  16. Waste reduction process improvements in the analysis of plutonium by x-ray fluorescence: results from multiple data sets

    Energy Technology Data Exchange (ETDEWEB)

    Worley, Christopher G [Los Alamos National Laboratory; Soderberg, Constance B [Los Alamos National Laboratory; Townsend, Lisa E [Los Alamos National Laboratory

    2010-01-01

    To minimize waste, improve process safety, and minimize costs, modifications were implemented to a method for quantifying gallium in plutonium metal using wavelength dispersive X-ray fluorescence. These changes included reducing sample sizes, reducing ion exchange process volumes, using cheaper reagent grade acids, eliminating the use of HF acid, and using more robust containment film for sample analysis. Relative precision and accuracy achieved from analyzing multiple aliquots from a single parent sample were {approx}0.2% and {approx}0.1% respectively. The same precision was obtained from analyzing a total of four parent materials, and the average relative accuracy from all the samples was 0.4%, which is within programmatic uncertainty requirements.

  17. The complementary roles of surface-immunoglobulin clonality and fluorescence intensity, mouse rosettes, and cd5 in the diagnosis of B-chronic lymphocytic-leukemia.

    Science.gov (United States)

    Batata, A; Shen, B; Yanes, B; Nicholson, G; Hines, D; Chang, J; Gross, H; Llenadolee, M; Merle, S

    1993-03-01

    Peripheral blood from 77 cases of B-CLL was analyzed to evaluate the diagnostic value of SIg clonality and fluorescence intensity, mouse rosettes (MR), and CD5. Monoclonal SIg (L-chain restriction or H-chain restriction or both) was detected in 69 cases (89.61%), with weak fluorescence (mean channel number on flow cytometry <200) in 65 (94.2%); MR was positive in 66 (85.71%); CD5 positive in 64 (83.12%). The association of SIg intensity, MR, and CD5 was as follows: weak SIg/MR+/CD5+, 41 cases (53.25%); weak SIg/MR+/CD5-, 13 (16.88%); weak SIg/MR-/CD5+, 11 (14.29%); strong SIg/MR+/CD5+, 4 (5.19%); and undetected SIg/MR+/CD5+, 8 (10.39%). Thus, by performing the three markers and accepting either two or three positive results (weak SIg, MR+, CD5+) as sufficient for diagnosis, all 77 cases (100%) were diagnosed. The study demonstrated the complementary roles between L- and H- chains, and between SIg intensity, MR, and CD5 in immunophenotyping CLL.

  18. Correlation of tryptophan fluorescence intensity decay parameters with sup 1 H NMR-determined rotamer conformations: (tryptophan sup 2 )oxytocin

    Energy Technology Data Exchange (ETDEWEB)

    Ross, J.B.A.; Schwartz, G.P.; Laws, W.R. (Mount Sinai, New York, NY (United States)); Wyssbrod, H.R.; Porter, R.A. (Univ. of Louisville, KY (United States)); Michaels, C.A. (Swarthmore Coll., PA (United States))

    1992-02-18

    While the fluorescence decay kinetics of tyrosine model compounds can be explained in terms of heterogeneity derived from the three ground-state {chi}{sup 1} rotamers, a similar correlation has yet to be directly observed for a tryptophan residue. In addition, the asymmetric indole ring might also lead to heterogeneity from {chi}{sup 2} rotations. In this paper, the time-resolved and steady-state fluorescence properties of (tryptophan{sup 2})oxytocin at pH 3 are presented and compared with {sup 1}H NMR results. According to the unrestricted analyses of individual fluorescence decay curves taken as a function of emission wavelength-independent decay constants, only three exponential terms are required. In addition, the preexponential weighting factors (amplitudes) have the same relative relationship (weights) as the {sup 1}H NMR-determined {chi}{sup 1} rotamer populations of the indole side chain. {sup 15}N was used in heteronuclear coupling experiments to confirm the rotamer assignments. Inclusion of a linked function restricting the decay amplitudes to the {chi}{sup 1} rotamer populations in the individual decay curve analyses and in the global analysis confirms this correlation. According to qualitative nuclear Overhauser data, there are two {chi}{sup 2} populations.

  19. Fluorescence excitation involving multiple electron transition states of N{sub 2} and CO{sub 2}

    Energy Technology Data Exchange (ETDEWEB)

    Wu, C.Y.R.; Chen, F.Z.; Hung, T.; Judge, D.L. [Univ. of Southern California, Los Angeles, CA (United States)

    1997-04-01

    The electronic states and electronic structures of N{sub 2} and CO{sub 2} in the 8-50 eV energy region have been studied extensively both experimentally and theoretically. In the energy region higher than 25 eV there exists many electronic states including multiple electron transition (MET) states which are responsible for producing most of the dissociative photoionization products. The electronic states at energies higher than 50 eV have been mainly determined by Auger spectroscopy, double charge transfer, photofragment spectroscopy and ion-ion coincidence spectroscopy. The absorption and ionization spectra of these molecules at energies higher than 50 eV mainly show a monotonic decrease in cross section values and exhibit structureless features. The decay channels of MET and Rydberg (or superexcited) states include autoionization, ionization, dissociative ionization, predissociation, and dissociation while those of single ion and multiple ion states may involve predissociation. and dissociation processes. The study of fluorescence specifically probes electronically excited species resulting from the above-mentioned decay channels and provides information for understanding the competition among these channels.

  20. Pointwise mutual information quantifies intratumor heterogeneity in tissue sections labeled with multiple fluorescent biomarkers

    Directory of Open Access Journals (Sweden)

    Daniel M Spagnolo

    2016-01-01

    Full Text Available Background: Measures of spatial intratumor heterogeneity are potentially important diagnostic biomarkers for cancer progression, proliferation, and response to therapy. Spatial relationships among cells including cancer and stromal cells in the tumor microenvironment (TME are key contributors to heterogeneity. Methods: We demonstrate how to quantify spatial heterogeneity from immunofluorescence pathology samples, using a set of 3 basic breast cancer biomarkers as a test case. We learn a set of dominant biomarker intensity patterns and map the spatial distribution of the biomarker patterns with a network. We then describe the pairwise association statistics for each pattern within the network using pointwise mutual information (PMI and visually represent heterogeneity with a two-dimensional map. Results: We found a salient set of 8 biomarker patterns to describe cellular phenotypes from a tissue microarray cohort containing 4 different breast cancer subtypes. After computing PMI for each pair of biomarker patterns in each patient and tumor replicate, we visualize the interactions that contribute to the resulting association statistics. Then, we demonstrate the potential for using PMI as a diagnostic biomarker, by comparing PMI maps and heterogeneity scores from patients across the 4 different cancer subtypes. Estrogen receptor positive invasive lobular carcinoma patient, AL13-6, exhibited the highest heterogeneity score among those tested, while estrogen receptor negative invasive ductal carcinoma patient, AL13-14, exhibited the lowest heterogeneity score. Conclusions: This paper presents an approach for describing intratumor heterogeneity, in a quantitative fashion (via PMI, which departs from the purely qualitative approaches currently used in the clinic. PMI is generalizable to highly multiplexed/hyperplexed immunofluorescence images, as well as spatial data from complementary in situ methods including FISSEQ and CyTOF, sampling many different

  1. A fluorescence-based centrifugal microfluidic system for parallel detection of multiple allergens

    Science.gov (United States)

    Chen, Q. L.; Ho, H. P.; Cheung, K. L.; Kong, S. K.; Suen, Y. K.; Kwan, Y. W.; Li, W. J.; Wong, C. K.

    2010-02-01

    This paper reports a robust polymer based centrifugal microfluidic analysis system that can provide parallel detection of multiple allergens in vitro. Many commercial food products (milk, bean, pollen, etc.) may introduce allergy to people. A low-cost device for rapid detection of allergens is highly desirable. With this as the objective, we have studied the feasibility of using a rotating disk device incorporating centrifugal microfluidics for performing actuationfree and multi-analyte detection of different allergen species with minimum sample usage and fast response time. Degranulation in basophils or mast cells is an indicator to demonstrate allergic reaction. In this connection, we used acridine orange (AO) to demonstrate degranulation in KU812 human basophils. It was found that the AO was released from granules when cells were stimulated by ionomycin, thus signifying the release of histamine which accounts for allergy symptoms [1-2]. Within this rotating optical platform, major microfluidic components including sample reservoirs, reaction chambers, microchannel and flow-control compartments are integrated into a single bio-compatible polydimethylsiloxane (PDMS) substrate. The flow sequence and reaction time can be controlled precisely. Sequentially through varying the spinning speed, the disk may perform a variety of steps on sample loading, reaction and detection. Our work demonstrates the feasibility of using centrifugation as a possible immunoassay system in the future.

  2. Influence of phase transitions on green fluorescence intensity ratio in Er3+ doped Ksub>0.5sub>Nasub>0.5sub>NbOsub>3sub> ceramic.

    Science.gov (United States)

    Liang, Zhang; Sun, Enwei; Pei, Shenghai; Li, Leipeng; Qin, Feng; Zheng, Yangdong; Zhao, Hua; Zhang, Zhiguo; Cao, Wenwu

    2016-12-12

    The fluorescence intensity ratio (FIR) method is a non-contact temperature (T) measurement technique based on thermally coupled levels of rare earth ions in a doped host. Green fluorescence originating from 2Hsub>11/2sub> and 4Ssub>3/2sub> states of Er3+ doped Ksub>0.5sub>Nasub>0.5sub>NbOsub>3sub> (KNN) ceramic are studied in the temperature range of 300 K to 720 K. The fluorescence intensities change dramatically around phase transition points where the crystal symmetry changes, inducing deviation of the FIR from Boltzmann's law. The temperature determined by the FIR method deviates from thermocouple measurements by 7 K at the orthorhombic to tetragonal phase transition (Tsub>O-Tsub>) point and 13 K at the Curie point (Tsub>Csub>). This finding gives guidance for developing fluorescent T sensors with ferroelectrics and may also provide a fluorescent method to detect phase transitions in ferroelectric materials.

  3. Double subgenomic alphaviruses expressing multiple fluorescent proteins using a Rhopalosiphum padi virus internal ribosome entry site element.

    Directory of Open Access Journals (Sweden)

    Michael R Wiley

    Full Text Available Double subgenomic Sindbis virus (dsSINV vectors are widely used for the expression of proteins, peptides, and RNA sequences. These recombinant RNA viruses permit high level expression of a heterologous sequence in a wide range of animals, tissues, and cells. However, the alphavirus genome structure and replication strategy is not readily amenable to the expression of more than one heterologous sequence. The Rhopalosiphum padi virus (RhPV genome contains two internal ribosome entry site (IRES elements that mediate cap-independent translation of the virus nonstructural and structural proteins. Most IRES elements that have been characterized function only in mammalian cells but previous work has shown that the IRES element present in the 5' untranslated region (UTR of the RhPV genome functions efficiently in mammalian, insect, and plant systems. To determine if the 5' RhPV IRES element could be used to express more than one heterologous sequence from a dsSINV vector, RhPV 5' IRES sequences were placed between genes for two different fluorescent marker proteins in the dsSINV, TE/3'2J/mcs. While mammalian and insect cells infected with recombinant viruses containing the RhPV sequences expressed both fluorescent marker proteins, only single marker proteins were routinely observed in cells infected with dsSINV vectors in which the RhPV IRES had been replaced by a luciferase fragment, an antisense RhPV IRES, or no intergenic sequence. Thus, we report development of a versatile tool for the expression of multiple sequences in diverse cell types.

  4. Novel lanthanide pH fluorescent probes based on multiple emissions and its visible-light-sensitized feature

    Energy Technology Data Exchange (ETDEWEB)

    Lin, Jintai [School of Chemistry and Environment, South China Normal University, Guangzhou 510006 (China); Zheng, Yuhui, E-mail: yhzheng78@scnu.edu.cn [School of Chemistry and Environment, South China Normal University, Guangzhou 510006 (China); Wang, Qianming, E-mail: qmwang@scnu.edu.cn [Key Laboratory of Theoretical Chemistry of Environment, Ministry of Education, School of Chemistry and Environment, South China Normal University, Guangzhou 510006 (China); School of Chemistry and Environment, South China Normal University, Guangzhou 510006 (China); Guangzhou Key Laboratory of Materials for Energy Conversion and Storage, Guangzhou 510006 (China); State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082 (China); Zeng, Zhi [School of Chemistry and Environment, South China Normal University, Guangzhou 510006 (China); Zhang, Cheng Cheng [Departments of Physiology and Developmental Biology, University of Texas Southwestern Medical Center, Dallas (United States)

    2014-08-11

    Graphical abstract: A new type of Eu(III) ofloxacin complex as the fluorescent pH indicator has been reported. Compared to pure ligand, the complex offers more distinguished color changes (green–red–blue) derived from both lanthanide line emissions and the secondary ionization steps of ofloxacin. - Highlights: • The pH probe offers a very wide working range in water (pH 1–14). • The emission changes have multiple colors. • Long-lived excited state lifetimes of Eu(III) has been used. • Two types of pH sensitive hydrogels were fabricated. - Abstract: A new type of Eu(III) ofloxacin complex as the fluorescent pH indicator has been presented. Compared to pure ligand, the complex offers more distinguished color changes (green–red–blue) derived from both lanthanide line emissions and the secondary ionization steps of ofloxacin. During the concentration dependence experiments, the photoluminescence studies on the complex showed that the excitation of this pH probe can occur at a very long wavelength which extends to visible range (Ex = 427 nm). Furthermore, the functional complex was successfully incorporated into soft networks and two novel luminescent hydrogels (rod and film) were fabricated. The soft materials also exhibited specific responses towards the pH variation. Finally, the onion cell-stain experiments were carried out to further confirm the validity of pH dependence and the results support the idea that the material will be suitable for monitoring biological samples in the future.

  5. High Intensity Exercise in Multiple Sclerosis: Effects on Muscle Contractile Characteristics and Exercise Capacity, a Randomised Controlled Trial.

    Directory of Open Access Journals (Sweden)

    Inez Wens

    Full Text Available Low-to-moderate intensity exercise improves muscle contractile properties and endurance capacity in multiple sclerosis (MS. The impact of high intensity exercise remains unknown.Thirty-four MS patients were randomized into a sedentary control group (SED, n = 11 and 2 exercise groups that performed 12 weeks of a high intensity interval (HITR, n = 12 or high intensity continuous cardiovascular training (HCTR, n = 11, both in combination with resistance training. M.vastus lateralis fiber cross sectional area (CSA and proportion, knee-flexor/extensor strength, body composition, maximal endurance capacity and self-reported physical activity levels were assessed before and after 12 weeks.Compared to SED, 12 weeks of high intensity exercise increased mean fiber CSA (HITR: +21 ± 7%, HCTR: +23 ± 5%. Furthermore, fiber type I CSA increased in HCTR (+29 ± 6%, whereas type II (+23 ± 7% and IIa (+23 ± 6%, CSA increased in HITR. Muscle strength improved in HITR and HCTR (between +13 ± 7% and +45 ± 20% and body fat percentage tended to decrease (HITR: -3.9 ± 2.0% and HCTR: -2.5 ± 1.2%. Furthermore, endurance capacity (Wmax +21 ± 4%, time to exhaustion +24 ± 5%, VO2max +17 ± 5% and lean tissue mass (+1.4 ± 0.5% only increased in HITR. Finally self-reported physical activity levels increased 73 ± 19% and 86 ± 27% in HCTR and HITR, respectively.High intensity cardiovascular exercise combined with resistance training was safe, well tolerated and improved muscle contractile characteristics and endurance capacity in MS.ClinicalTrials.gov NCT01845896.

  6. Intensity dependent absorption bleaching of high subband excitons in GaAs/AlGaAs multiple quantum wells

    CERN Document Server

    Shin, S H; Lee, E H; Chae, K M; Park, S H; Kim, U

    1998-01-01

    We have investigated the influence of carrier generation on the absorption bleaching of the n=2 and n=3 excitons in GaAs/AlGaAs multiple quantum wells (MQWs). With the excitation near the resonance of the n=1 exciton absorption, the long range coulomb screening and collision broadening had significant effects on the exciton bleaching. At low excitation intensity, the absorption bleaching of the n=2 exciton in 75 A-thick MQWs and that of the n=3 exciton in 150 A-thick MQWs were due to linewidth broadening by the collision broadening effect only. At high excitation intensity, however, the reduction of oscillator strength due to the long range coulomb screening contributed dominantly to absorption bleaching.

  7. Changes to processes in estuaries and coastal waters due to intense multiple pressures - An introduction and synthesis

    Science.gov (United States)

    Mitchell, Steven B.; Jennerjahn, Tim C.; Vizzini, Salvatrice; Zhang, Weiguo

    2015-04-01

    From the 2013 ECSA conference 'Estuaries and Coastal Areas in Times of Intense Change' a theme emerged that has ended up being the focus of this Special Issue of Estuarine Coastal and Shelf Science, namely 'Changes to processes in estuaries and coastal waters due to intense multiple pressures'. Many parts of the world are continuing to experience unprecedented rates of economic growth, and those responsible for managing coastal and estuarine areas must respond accordingly. At the same time, global climate change and sea level rise are also continuing, placing new or more intense pressures on coastal areas that must be dealt with in ways that are as far as possible managed as a result of good scientific understanding. There are other pressures too, which depend on the system concerned. This article provides an overview of the papers contained within the Special Issue and provides a discussion of how these fit within the main theme of intense multiple stressors, considering how a balance can be achieved between the needs of various different stakeholders and interest groups, and the sustainability of the system concerned. We categorise the papers in four main groupings: (1) stressors related to sea level rise; (2) stressors related to changes in fresh water inputs; (3) stressors related to anthropogenic pollution; and (4) the use of indicators as a means of assessing the effects of stressors, and reflect on the fact that despite the diversity of different challenges and geographical regions involved many of the approaches and discussions contained within the Special Issue have strong similarities, leading to a set of overarching principles that should be considered when making recommendations on management strategies.

  8. Comparison study of intensity modulated arc therapy using single or multiple arcs to intensity modulated radiation therapy for high-risk prostate cancer

    Energy Technology Data Exchange (ETDEWEB)

    Ashamalla, Hani; Tejwani, Ajay; Parameritis, Loannis; Swamy, Uma; Luo, Pei Ching; Guirguis, Adel; Lavaf, Amir [Weill Medical College of Cornell University, Brooklyn, NY (United States)

    2013-06-15

    Intensity modulated arc therapy (IMAT) is a form of intensity modulated radiation therapy (IMRT) that delivers dose in single or multiple arcs. We compared IMRT plans versus single-arc field (1ARC) and multi-arc fields (3ARC) IMAT plans in high-risk prostate cancer. Sixteen patients were studied. Prostate (PTV{sub P}), right pelvic (PTV{sub RtLN}) and left pelvic lymph nodes (PTV{sub LtLN}), and organs at risk were contoured. PTVP, PTV{sub RtLN}, and PTV{sub LtLN} received 50.40 Gy followed by a boost to PTV{sub B} of 28.80 Gy. Three plans were per patient generated: IMRT, 1ARC, and 3ARC. We recorded the dose to the PTV, the mean dose (D{sub MEAN}) to the organs at risk, and volume covered by the 50% isodose. Efficiency was evaluated by monitor units (MU) and beam on time (BOT). Conformity index (CI), Paddick gradient index, and homogeneity index (HI) were also calculated. Average Radiation Therapy Oncology Group CI was 1.17, 1.20, and 1.15 for IMRT, 1ARC, and 3ARC, respectively. The plans' HI were within 1% of each other. The D{sub MEAN} of bladder was within 2% of each other. The rectum D{sub MEAN} in IMRT plans was 10% lower dose than the arc plans (p < 0.0001). The GI of the 3ARC was superior to IMRT by 27.4% (p = 0.006). The average MU was highest in the IMRT plans (1686) versus 1ARC (575) versus 3ARC (1079). The average BOT was 6 minutes for IMRT compared to 1.3 and 2.9 for 1ARC and 3ARC IMAT (p < 0.05). For high-risk prostate cancer, IMAT may offer a favorable dose gradient profile, conformity, MU and BOT compared to IMRT.

  9. Validation of eGFP fluorescence intensity for testing in vitro cytotoxicity according to ISO 10993-5.

    Science.gov (United States)

    Miller, Felicitas; Hinze, Ulf; Chichkov, Boris; Leibold, Wolfgang; Lenarz, Thomas; Paasche, Gerrit

    2017-05-01

    ISO 10993-5 provides one of the accepted standards for testing the biotoxicity of new materials. All of the recommended test procedures rely upon the uptake or metabolism of dye by living cells. Results of direct contact tests can be potentially compromised by interaction or adsorption of the dye or its metabolic products. Therefore, the aim of the current study was to validate the use of the eGFP signal of transfected NIH-3T3 fibroblasts with the results of the MTT test in order to provide a test procedure that is very close to the ISO 10993-5 but has the advantage of not relying on the addition of dye. Our tests show that the MTT assay detects cytotoxicity in the eGFP NIH-3T3 cells at least as well as in the L929 cells. To facilitate the validation, we chose to integrate the fluorescence measurements into the MTT test procedure. To that end, an additional washing step was introduced. Additionally, medium without phenol red was used, resulting in a very high correlation of both measurements. Without these modifications, the fluorescence test was comparable to the MTT test in its ability to detect the cytotoxic potential of substances; however, it did result in slightly elevated IC50 concentrations. As the results of both tests correlated highly, measurement of the eGFP signal appears to present a reliable tool for detecting cytotoxicity of materials in line with the ISO 10993-5 norm with the advantage of avoiding the addition of dyes and the subsequent potential interaction with test materials. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 715-722, 2017. © 2015 Wiley Periodicals, Inc.

  10. Multiple magnetic relaxation processes, magnetocaloric effect and fluorescence properties of rhombus-shaped tetranuclear rare earth complexes.

    Science.gov (United States)

    Gao, Hong-Ling; Jiang, Li; Liu, Shuang; Shen, Hai-Yun; Wang, Wen-Min; Cui, Jian-Zhong

    2016-01-07

    Seven new tetranuclear rare earth (RE) complexes [RE4(acac)4L6(μ3-OH)2] (HL = 5-(4-fluorobenzylidene)-8-hydroxylquinoline; acac = acetylacetonate; RE = Y (1), Eu (2), Gd (3), Tb (4), Dy (5), Tm (6) and Lu (7)) have been synthesized and completely characterized. Complex exhibits multiple zero-field slow magnetic relaxation processes typical of Single Molecule Magnets (SMMs). Two distinct slow magnetic relaxation processes, with effective energy barriers of Ueff = 48 K for the slow relaxation (SR) process and Ueff = 121 K for the fast relaxation (FR) process, are mainly attributed to the presence of two crystallographically independent Dy(III) sites. The magnetocaloric effect (MCE) was detected as -ΔSm(T) = 20.8 J kg(-1) K(-1) for complex . The fluorescence properties of complexes 1, 2, 4, 5 and 7 were also investigated. Complexes 2, 4 and 5 show the characteristic peaks for their corresponding RE(III) center, while complexes 1 and 7 show similar emission peaks to the Schiff base ligand when they are excited at the appropriate wavelength.

  11. Bayesian Hierarchical Model for Estimating Gene Expression Intensity Using Multiple Scanned Microarrays

    Directory of Open Access Journals (Sweden)

    Auvinen Petri

    2008-01-01

    Full Text Available We propose a method for improving the quality of signal from DNA microarrays by using several scans at varying scanner sen-sitivities. A Bayesian latent intensity model is introduced for the analysis of such data. The method improves the accuracy at which expressions can be measured in all ranges and extends the dynamic range of measured gene expression at the high end. Our method is generic and can be applied to data from any organism, for imaging with any scanner that allows varying the laser power, and for extraction with any image analysis software. Results from a self-self hybridization data set illustrate an improved precision in the estimation of the expression of genes compared to what can be achieved by applying standard methods and using only a single scan.

  12. Intense red upconversion fluorescence emission in NIR-excited erbium-ytterbium doped laponite-derived phosphor

    Science.gov (United States)

    da Silva, Andréa F.; Moura, Diógenes S.; Gouveia-Neto, Artur S.; Silva, Elias A.; Bueno, Luciano A.; Costa, Ernande B.; Azevedo, Eduardo N.

    2011-09-01

    In this report the optical properties and energy-transfer frequency upconversion luminescence of Er 3+/Yb 3+-codoped laponite-derived powders under 975 nm infrared excitation is investigated. The 75%(laponite):25%(PbF 2) samples doped with erbium and ytterbium ions, generated high intensity red emission around 660 nm and lower intensity green emission around 525, and 545 nm. The observed emission signals were examined as a function of the excitation power and annealing temperature. The results indicate that energy-transfer, and excited-state absorption are the major upconversion excitation mechanism for the erbium excited-state red emitting level. The precursor glass samples were also heat treated at annealing temperatures of 300 °C, 400 °C, 500 °C, and 600 °C, for a 2 h period. The dependence of the visible upconversion luminescence emission upon the annealing temperature indicated the existence of an optimum temperature which leads to the generation of the most intense and spectrally pure red emission signal.

  13. Additive effects prevail: The response of biota to multiple stressors in an intensively monitored watershed.

    Science.gov (United States)

    Gieswein, Alexander; Hering, Daniel; Feld, Christian K

    2017-09-01

    Freshwater ecosystems are impacted by a range of stressors arising from diverse human-caused land and water uses. Identifying the relative importance of single stressors and understanding how multiple stressors interact and jointly affect biology is crucial for River Basin Management. This study addressed multiple human-induced stressors and their effects on the aquatic flora and fauna based on data from standard WFD monitoring schemes. For altogether 1095 sites within a mountainous catchment, we used 12 stressor variables covering three different stressor groups: riparian land use, physical habitat quality and nutrient enrichment. Twenty-one biological metrics calculated from taxa lists of three organism groups (fish, benthic invertebrates and aquatic macrophytes) served as response variables. Stressor and response variables were subjected to Boosted Regression Tree (BRT) analysis to identify stressor hierarchy and stressor interactions and subsequently to Generalised Linear Regression Modelling (GLM) to quantify the stressors standardised effect size. Our results show that riverine habitat degradation was the dominant stressor group for the river fauna, notably the bed physical habitat structure. Overall, the explained variation in benthic invertebrate metrics was higher than it was in fish and macrophyte metrics. In particular, general integrative (aggregate) metrics such as % Ephemeroptera, Plecoptera and Trichoptera (EPT) taxa performed better than ecological traits (e.g. % feeding types). Overall, additive stressor effects dominated, while significant and meaningful stressor interactions were generally rare and weak. We concluded that given the type of stressor and ecological response variables addressed in this study, river basin managers do not need to bother much about complex stressor interactions, but can focus on the prevailing stressors according to the hierarchy identified. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. EFFECT OF Cu AND Mn TOXICITY ON CHLOROPHYLL FLUORESCENCE AND GAS EXCHANGE IN RICE AND SUNFLOWER UNDER DIFFERENT LIGHT INTENSITIES

    Directory of Open Access Journals (Sweden)

    Hajiboland R.

    2007-06-01

    Full Text Available Copper (Cu and manganese (Mn are essential micronutrients for plants, but toxic at high concentrations. Responses of rice (Oryza sativa L. and sunflower (Helianthus annuus L. to toxic concentrations of Mn and Cu (up to 100 μM were studied under three light intensities including low (LL, PPFD=100, intermediate (IL, PPFD=500 and high (HL, PPFD=800 light intensities in hydroponic medium. Rice plants showed higher susceptibility than sunflower to both heavy metals concerning dry matter of shoot and root. Growing under higher light intensity strengthened the effect of Cu toxicity while ameliorated that of Mn, the latter was attributed to the lower Mn accumulation of HL plants in both shoot and root. Chlorophyll content of leaves was influenced negatively only by Cu treatment and that at the highest concentration in the medium (100 μM. Similar with growth results, reduction of net assimilation rate (A was higher in HL than LL plants treated by excess Cu, but in contrast to growth response, reduction was more prominent in sunflower than rice. Excess Mn-induced reduction of A was similar between LL and HL plants and was greater in sunflower than rice. Reduction of A was partly attributable to stomatal limitation, but non-stomatal mechanisms were also involved in this reduction. Copper and Mn treatment did not change the optimal quantum efficiency of PSII in dark-adapted chloroplasts (Fv/Fm ratio, but Fv/F0 was influenced particularly by Cu treatment, the reduction was higher in rice than sunflower and in HL compared to LL plants. Regarding excess Cu and Mn-mediated alterations in chlorophyll concentration, Fv/F0 and Tm values, it was suggested that, Cu and Mn toxicity depress the leaf photosynthetic capacity primarily by causing a significant alteration of the composition and functional competence of the photosynthetic units rather a reduction in the number of photosynthetic units (PSUs per unit leaf area.

  15. Land-Use Intensity of Electricity Production: Comparison Across Multiple Sources

    Science.gov (United States)

    Swain, M.; Lovering, J.; Blomqvist, L.; Nordhaus, T.; Hernandez, R. R.

    2015-12-01

    Land is an increasingly scarce global resource that is subject to competing pressures from agriculture, human settlement, and energy development. As countries concerned about climate change seek to decarbonize their power sectors, renewable energy sources like wind and solar offer obvious advantages. However, the land needed for new energy infrastructure is also an important environmental consideration. The land requirement of different electricity sources varies considerably, but there are very few studies that offer a normalized comparison. In this paper, we use meta-analysis to calculate the land-use intensity (LUI) of the following electricity generation sources: wind, solar photovoltaic (PV), concentrated solar power (CSP), hydropower, geothermal, nuclear, biomass, natural gas, and coal. We used data from existing studies as well as original data gathered from public records and geospatial analysis. Our land-use metric includes land needed for the generation facility (e.g., power plant or wind farm) as well as the area needed to mine fuel for natural gas, coal, and nuclear power plants. Our results found the lowest total LUI for nuclear power (115 ha/TWh/y) and the highest LUI for biomass (114,817 ha/TWh/y). Solar PV and CSP had a considerably lower LUI than wind power, but both were an order of magnitude higher than fossil fuels (which ranged from 435 ha/TWh/y for natural gas to 579 ha/TWh/y for coal). Our results suggest that a large build-out of renewable electricity, though it would offer many environmental advantages over fossil fuel power sources, would require considerable land area. Among low-carbon energy sources, relatively compact sources like nuclear and solar have the potential to reduce land requirements.

  16. Health-related quality of life and rehabilitation cost following intensive care unit stay in multiple trauma patients.

    Science.gov (United States)

    Stergiannis, Pantelis; Katsoulas, Theodoros; Fildissis, George; Intas, George; Galanis, Peter; Kosta, Natalia; Zidianakis, Vasilios; Baltopoulos, George

    2014-01-01

    The objective of this study was to assess changes in health-related quality of life (HRQOL) in multiple trauma patients due to motor vehicle crashes during a follow-up period of 2 years after discharge from an intensive care unit (ICU) and the effect of income and financial cost of rehabilitation in HRQOL. The study was a prospective observational study of multiple trauma patients from January 2009 to January 2011 who were hospitalized in a general, medical, and surgical ICU of a district hospital in Athens, Greece. Eighty-five patients with multiple traumas due to motor vehicle crashes and with an ICU stay of more than 24 hours were included in the study. HRQOL was assessed by a general questionnaire, the EuroQol 5D. Increased monthly household income and absence of traumatic brain injuries were associated with an improved EQ-VAS score. The frequency of severe problems in mobility, self-care, usual activities, pain/discomfort, and anxiety/depression decreased over time. The financial cost of rehabilitation was initially high but decreased over time. Severely injured victims of motor vehicle crashes suffer from serious problems in terms of HRQOL which is gradually improved even 2 years after hospital discharge. In addition, HRQOL is significantly related to income. Resources used for rehabilitation decrease over time, but even at 24 months, the patients still use half of the amount as compared with the cost of the first 6 months after trauma.

  17. Detection of Inter-chromosomal Stable Aberrations by Multiple Fluorescence In Situ Hybridization (mFISH) and Spectral Karyotyping (SKY) in Irradiated Mice.

    Science.gov (United States)

    Pathak, Rupak; Koturbash, Igor; Hauer-Jensen, Martin

    2017-01-11

    Ionizing radiation (IR) induces numerous stable and unstable chromosomal aberrations. Unstable aberrations, where chromosome morphology is substantially compromised, can easily be identified by conventional chromosome staining techniques. However, detection of stable aberrations, which involve exchange or translocation of genetic materials without considerable modification in the chromosome morphology, requires sophisticated chromosome painting techniques that rely on in situ hybridization of fluorescently labeled DNA probes, a chromosome painting technique popularly known as fluorescence in situ hybridization (FISH). FISH probes can be specific for whole chromosome/s or precise sub-region on chromosome/s. The method not only allows visualization of stable aberrations, but it can also allow detection of the chromosome/s or specific DNA sequence/s involved in a particular aberration formation. A variety of chromosome painting techniques are available in cytogenetics; here two highly sensitive methods, multiple fluorescence in situ hybridization (mFISH) and spectral karyotyping (SKY), are discussed to identify inter-chromosomal stable aberrations that form in the bone marrow cells of mice after exposure to total body irradiation. Although both techniques rely on fluorescent labeled DNA probes, the method of detection and the process of image acquisition of the fluorescent signals are different. These two techniques have been used in various research areas, such as radiation biology, cancer cytogenetics, retrospective radiation biodosimetry, clinical cytogenetics, evolutionary cytogenetics, and comparative cytogenetics.

  18. Deployment and operation of a transportable burn intensive care unit in response to a burn multiple casualty incident.

    Science.gov (United States)

    Barillo, David J; Cancio, Leopoldo C; Stack, Richard S; Carr, Shamus R; Broger, Kristine P; Crews, David M; Renz, Evan M; Blackbourne, Lorne H

    2010-01-01

    In many hospitals, intensive care units (ICUs) operate at or above capacity on a daily basis. Multiple casualty incidents will create a sudden need for additional ICU beds and hospital planning for disaster response must anticipate the need for rapid ICU expansion. In this article, the authors describe the management of 6 patients who were burned in Guam and successfully transported a distance of 7,268 miles to San Antonio, TX, for tertiary burn center care. The mission required creation of a temporary burn ICU at Tripler Army Medical Center in Hawaii, approximately midway between the referring hospital and the receiving burn center. A method of creating a temporary burn center is described. Lessons learned, including the need to standardize equipment, and to cross-train and cross-credential medical personnel, are applicable to both military and civilian mass casualty management.

  19. Stationary Intensity Distribution of Single-Mode Laser Driven by Additive and Multiplicative Colored Noises with Colored Cross-Correlation

    Institute of Scientific and Technical Information of China (English)

    LIANG Gui-Yun; CAO Li; WANG Jun; Wu Da-Jin

    2003-01-01

    Applying the approximate Fokker-Planck equation we derived, we obtain the analytic expression of thestationary laser intensity distribution Pst(Ⅰ) by studying the single-mode laser cubic model subject to colored cross-correlation additive and multiplicative noise, each of which is colored. Based on it, we discuss the effects on the stationarylaser intensity distribution Pst(Ⅰ) by cross-correlation between noises and "color" of noises (non-Markovian effect) whenthe laser system is above the threshold. In detail, we analyze two cases: One is that the three correlation-times (i.e.the self-correlation and cross-correlation times of the additive and multiplicative noise) are chosen to be the same value(τ1 = τ2 = τ3 = τ). For this case, the effect of noise cross-correlation is investigated emphatically, and we detect thatonly when λ≠ 0 can the noise-induced transition occur in the Pst(Ⅰ) curve, and only when τ≠ 0 and λ≠ 0, can the"reentrant noise-induced transition" occur. The other case is that the three correlation times are not the same value,τ1 ≠τ2 ≠τ3. For this case, we find that the noise-induced transition occurring in the Pst (Ⅰ) curve is entirely differentwhen the values of τ1, τ2, and τ3 are changed respectively. In particular, when τ2 (self-correlation time of additivenoise) is changing, the ratio of the two maximums of the Pst(Ⅰ) curve R exhibits an interesting phenomenon, "reentrantnoise-induced transition", which demonstrates the effect of noise "color" (non-Markovian effect).

  20. Stationary Intensity Distribution of Single-Mode Laser Driven by Additive and Multiplicative Colored Noises with Colored Cross-Correlation

    Institute of Scientific and Technical Information of China (English)

    CAOLi; WANGJun; WuDa-Jin; LIANGGui-Yun

    2003-01-01

    Applying the approximate Fokker-Planck equation we derived, we obtain the analytic expression of the stationary laser intensity distribution Pst(l) by studying the single-mode laser cubic model subject to colored cross-correlation additive and multiplicative noise, each of which is colored. Based on it, we discuss the effects on the stationary laser intensity distribution Pot(I) by cross-correlation between noises and "color" of noises (non-Markovian effect) when the laser system is above the threshold. In detail, we analyze two cases: One is that the three correlation-times (i.e.the self-correlation and cross-correlation times of the additive and multiplicative noise) are chosen to be the same value(Tl=T2=T3=T). For this case, the effect of noise cross-correlation is investigated emphatically, and we detect that only when λ≠ 0 can the noise-induced transition occur in the Pst (I) curve, and only when T≠ 0 and λ≠0, can the "reentrant noise-induced transition" occur. The other case is that the three correlation times are not the same value,T1≠T2≠T3. For this case, we find that the noise-induced transition occurring in the Pst(I) curve is entirely different when the values of T1,T2, and T3 are changed respectively. In particular, when T2 (serf-correlation time of additive noise) is cha~g~g, the ratio of the two maximums of the Pst( I) curve R exhibits an interesting phenomenon,"reentrant noise-induced transition", which demonstrates the effect of noise "color" (non-Markovian effect).

  1. Effects of temperature, CO 2/O 2 concentrations and light intensity on cellular multiplication of microalgae, Euglena gracilis

    Science.gov (United States)

    Kitaya, Y.; Azuma, H.; Kiyota, M.

    Microalgae culture is likely to play an important role in aquatic food production modules in bioregenerative systems for producing feeds for fish, converting CO 2 to O 2 and remedying water quality as well as aquatic higher plants. In the present study, the effects of culture conditions on the cellular multiplication of microalgae, Euglena gracilis, was investigated as a fundamental study to determine the optimum culture conditions for microalgae production in aquatic food production modules including both microalgae culture and fish culture systems. E. gracilis was cultured under conditions with five levels of temperatures (25-33 °C), three levels of CO 2 concentrations (2-6%), five levels of O 2 concentrations (10-30%), and six levels of photosynthetic photon flux (20-200 μmol m -2 s -1). The number of Euglena cells in a certain volume of solution was monitored with a microscope under each environmental condition. The multiplication rate of the cells was highest at temperatures of 27-31 °C, CO 2 concentration of 4%, O 2 concentration of 20% and photosynthetic photon flux of about 100 μmol m -2 s -1. The results demonstrate that E. gracilis could efficiently produce biomass and convert CO 2 to O 2 under relatively low light intensities in aquatic food production modules.

  2. Effects of temperature, CO2/O2 concentrations and light intensity on cellular multiplication of microalgae, Euglena gracilis

    Science.gov (United States)

    Kitaya, Y.; Azuma, H.; Kiyota, M.

    2005-01-01

    Microalgae culture is likely to play an important role in aquatic food production modules in bioregenerative systems for producing feeds for fish, converting CO2 to O2 and remedying water quality as well as aquatic higher plants. In the present study, the effects of culture conditions on the cellular multiplication of microalgae, Euglena gracilis, was investigated as a fundamental study to determine the optimum culture conditions for microalgae production in aquatic food production modules including both microalgae culture and fish culture systems. E. gracilis was cultured under conditions with five levels of temperatures (25-33 degrees C), three levels of CO2 concentrations (2-6%), five levels of O2 concentrations (10-30%), and six levels of photosynthetic photon flux (20-200 micromoles m-2 s-1). The number of Euglena cells in a certain volume of solution was monitored with a microscope under each environmental condition. The multiplication rate of the cells was highest at temperatures of 27-31 degrees C, CO2 concentration of 4%, O2 concentration of 20% and photosynthetic photon flux of about 100 micromoles m-2 s-1. The results demonstrate that E. gracilis could efficiently produce biomass and convert CO2 to O2 under relatively low light intensities in aquatic food production modules. c2005 Published by Elsevier Ltd on behalf of COSPAR.

  3. Neutrophil and Monocyte CD64 and CD163 Expression in Critically Ill Neonates and Children with Sepsis: Comparison of Fluorescence Intensities and Calculated Indexes

    Directory of Open Access Journals (Sweden)

    Mojca Groselj-Grenc

    2008-01-01

    Full Text Available Objective. To evaluate the expression of CD64 and CD163 on neutrophils and monocytes in SIRS with/without sepsis and to compare the diagnostic accuracy of CD64 and CD163 molecules expression determined as (1 mean fluorescence intensities (MFI of CD64 and CD163; and (2 the ratio (index of linearized MFI to the fluorescence signal of standardized beads. Patients and methods. Fifty-six critically ill neonates and children with systemic inflammatory response syndrome (SIRS and suspected sepsis, classified into two groups: SIRS with sepsis (n=29 and SIRS without sepsis (n=27. Results. CD64 and CD163 MFI measured on neutrophils and monocytes were elevated in patients with SIRS with sepsis. Diagnostic accuracy of indexes was equal to diagnostic accuracy of MFI for CD64 on neutrophils (0.833 versus 0.854 for day 0 and 0.975 versus 0.983 for day 1 and monocytes (0.811 versus 0.865 for day 0 and 0.825 versus 0.858 for day 1, and CD163 on neutrophils (0.595 versus 0.655 for day 0 and 0.677 versus 0.750 for day 1, but not for CD163 on monocytes. Conclusion. CD64 MFI, CD163 MFI, CD64 indexes for neutrophils and monocytes, and CD163 index for neutrophils can all be used for discrimination of SIRS and sepsis in critically ill neonates and children. CD64 index for neutrophils, however, is superior to all other markers.

  4. Using a low-order model to detect and characterize intense vortices in multiple-Doppler radar data

    Science.gov (United States)

    Potvin, Corey Keith

    A new multiple-Doppler radar analysis technique is presented for the objective detection and characterization of intense vortices. The technique consists of fitting radial wind data from two or more radars to a simple analytical model of a vortex and its near-environment. The model combines a uniform flow, linear shear flow, linear divergence flow (all of which comprise a broadscale flow), and modified combined Rankine vortex. The vortex and its environment are allowed to translate. A cost-function accounting for the discrepancy between the model and observed radial winds is evaluated over space and time so that observations can be used at the actual times and locations they were acquired. The parameters in the low-order model are determined by minimizing this cost function. The development of the method is initially guided by emulated radial velocity observations of analytical vortices. A high-resolution Advanced Regional Prediction System (ARPS) simulation of a supercellular tornado is then used to generate more realistic pseudo-observations. Finally, the technique is tested using real dual-Doppler tornado and mesocyclone observations from a variety of radar platforms including Weather Surveillance Radar - 1988 Doppler (WSR-88D), Terminal Doppler Weather Radar (TDWR), Shared Mobile Atmospheric Research and Teaching Radar (SMART-R), and Doppler on Wheels (DOW). The technique shows skill in detecting intense vortices and, when the vortex is well-resolved, in retrieving key model parameters including vortex location, translational velocity, radius and maximum tangential wind speed. In cases where the vortex is not well-resolved, additional vortex characteristics computed from the retrieved model parameters and verified against radial velocity observations can still provide useful information about vortex size and strength.

  5. High intensity training may reverse the fiber type specific decline in myogenic stem cells in multiple sclerosis patients

    Directory of Open Access Journals (Sweden)

    Jean eFarup

    2016-05-01

    Full Text Available Multiple sclerosis (MS is associated with loss of skeletal muscle mass and function. The myogenic stem cells (satellite cells – SCs are instrumental to accretion of myonuclei, but remain to be investigated in MS. The present study aimed to compare the SC and myonuclei content between MS patients (n=23 and age matched healthy controls (HC, n=18. Furthermore, the effects of 12 weeks of high intensity training on SC and myonuclei content were explored in MS. Muscle biopsies were obtained from m. Vastus Lateralis at baseline (MS+HC and following 12 weeks of training (MS only. Frozen biopsies were sectioned followed by immunohistochemical analysis for fiber type specific SCs (Pax7+, myonuclei (MN and central nuclei content and fiber cross-sectional area (fCSA using ATPase histochemistry. At baseline the SCs per fiber was lower in type II compared to type I fiber in both MS (119%, p<0.01 and HC (69%, p<0.05, whereas the SCs per fCSA was lower in type II fibers compared to type I only in MS (72%, p<0.05. No differences were observed in MN or central nuclei between MS and HC. Following training the type II fiber SCs per fiber and fCSA in MS patients increased by 165% (p<0.05 and 135% (p<0.05, respectively. Furthermore, the type II fiber MN content increased by 35% (p<0.05 following training. In conclusion, the SC content is lower in type II compared to type I fibers in both MS and HC. Furthermore, high intensity training was observed to selectively increase the SC and myonuclei content in type II fibers in MS patients.

  6. Expanding the potential of standard flow cytometry by extracting fluorescence lifetimes from cytometric pulse shifts

    National Research Council Canada - National Science Library

    Cao, Ruofan; Naivar, Mark A; Wilder, Mark; Houston, Jessica P

    2014-01-01

    Fluorescence lifetime measurements provide information about the fluorescence relaxation, or intensity decay, of organic fluorophores, fluorescent proteins, and other inorganic molecules that fluoresce...

  7. Fate of biopolymers during rapeseed meal and wheat bran composting as studied by two-dimensional correlation spectroscopy in combination with multiple fluorescence labeling techniques.

    Science.gov (United States)

    Wang, Li-Ping; Shen, Qi-Rong; Yu, Guang-Hui; Ran, Wei; Xu, Yang-Chun

    2012-02-01

    Detailed knowledge of the molecular events during composting is important in improving the efficiency of this process. By combining two-dimensional Fourier transform infrared (FTIR) correlation spectroscopy and multiple fluorescent labeling, it was possible to study the degradation of biopolymers during rapeseed meal and wheat bran composting. Two-dimensional FTIR correlation spectroscopy provided structural information and was used to deconvolute overlapping bands found in the compost FTIR spectra. The degradation of biopolymers in rapeseed meal and wheat bran composts followed the sequence: cellulose, heteropolysaccharides, and proteins. Fluorescent labeling suggested that cellulose formed an intact network-like structure and the other biopolymers were embedded in the core of this structure. The sequence of degradation of biopolymers during composting was related to their distribution patterns.

  8. Continuous ultra-low-intensity artificial daylight is not as effective as red LED light in photodynamic therapy of multiple actinic keratoses

    DEFF Research Database (Denmark)

    Wiegell, Stine Regin; Heydenreich, Jakob; Fabricius, Susanne;

    2011-01-01

    Daylight-mediated photodynamic therapy (PDT) is a simple and tolerable treatment of nonmelanoma skin cancer. It is of interest which light intensity is sufficient to prevent accumulation of protoporphyrin IX (PpIX) and effectively treat actinic keratoses (AKs). We compared the efficacy of PDT...... with light-emitting diode (LED) to daylight-mediated PDT with very low-intensity artificial daylight ('daylight') in the treatment of multiple AKs in the face or scalp....

  9. The Cyan Fluorescent Protein (CFP) Transgenic Mouse as a Model for Imaging Pancreatic Exocrine Cells

    Science.gov (United States)

    Cao, Hop S Tran; Kimura, Hiroaki; Kaushal, Sharmeela; Snyder, Cynthia S; Reynoso, Jose; Hoffman, Robert M; Bouvet, Michael

    2015-01-01

    Context The use of fluorescent proteins for in vivo imaging has opened many new areas of research. Among the important advances in the field have been the development of transgenic mice expressing various fluorescent proteins. Objective To report whole-body and organ-specific fluorescence imaging to characterize the transgenic cyan fluorescent protein mouse. Design Mice were imaged using two devices. Brightfield images were obtained with the OV100 Small Animal Imaging System (Olympus Corp., Tokyo, Japan). Fluorescence imaging was performed under the cyan fluorescent protein filter using the iBox Small Animal Imaging System (UVP, Upland, CA, USA). Intervention All animals were sacrificed immediately before imaging. They were imaged before and throughout multiple steps of a complete necropsy. Harvested organs were also imaged with both devices. Selected organs were then frozen and processed for histology, fluorescence microscopy, and H&E staining. Fluorescence microscopy was performed with an Olympus IMT-2 inverted fluorescence microscope. Main outcome measure Determination of fluorescence intensity of different organs. Results Surprisingly, we found that there is differential enhancement of fluorescence among organs; most notably, the pancreas stands out from the rest of the gastrointestinal tract, displaying the strongest fluorescence of all organs in the mouse. Fluorescence microscopy demonstrated that the cyan fluorescent protein fluorescence resided in the acinar cells of the pancreas and not the islet cells. Conclusions The cyan fluorescent protein mouse should lead to a deeper understanding of pancreatic function and pathology, including cancer. PMID:19287108

  10. The Cyan Fluorescent Protein (CFP Transgenic Mouse as a Model for Imaging Pancreatic Exocrine Cells

    Directory of Open Access Journals (Sweden)

    Hop S Tran Cao

    2009-03-01

    Full Text Available The use of fluorescent proteins for in vivo imaging has opened many new areas of research. Among the important advances in the field have been the development of transgenic mice expressing various fluorescent proteins. Objective To report whole-body and organ-specific fluorescence imaging to characterize the transgenic cyan fluorescent protein mouse. Design Mice were imaged using two devices. Brightfield images were obtained with the OV100 Small Animal Imaging System (Olympus Corp., Tokyo, Japan. Fluorescence imaging was performed under the cyan fluorescent protein filter using the iBox Small Animal Imaging System (UVP, Upland, CA, USA. Intervention All animals were sacrificed immediately before imaging. They were imaged before and throughout multiple steps of a complete necropsy. Harvested organs were also imaged with both devices. Selected organs were then frozen and processed for histology, fluorescence microscopy, and H&E staining. Fluorescence microscopy was performed with an Olympus IMT-2 inverted fluorescence microscope. Main outcome measure Determination of fluorescence intensity of different organs. Results Surprisingly, we found that there is differential enhancement of fluorescence among organs; most notably, the pancreas stands out from the rest of the gastrointestinal tract, displaying the strongest fluorescence of all organs in the mouse. Fluorescence microscopy demonstrated that the cyan fluorescent protein fluorescence resided in the acinar cells of the pancreas and not the islet cells. Conclusions The cyan fluorescent protein mouse should lead to a deeper understanding of pancreatic function and pathology, including cancer.

  11. Interphase fluorescence in situ hybridization in multiple myeloma and monoclonal gammopathy of undetermined significance without and with positive plasma cell identification

    DEFF Research Database (Denmark)

    Christensen, Jacob H; Abildgaard, Niels; Plesner, Torben

    2007-01-01

    Interphase fluorescence in-situ hybridization (i-FISH) was used to investigate 192 patients with multiple myeloma (MM; n = 182) and benign monoclonal gammopathy of undetermined significance (MGUS; n = 10). Of the 182 MM cases, 132 were investigated without and 50 with positive plasma cell......32. Of these, translocations t(4;14) constituted 9% and t(11;14), 20%. Finally, based on the small number of cytogenetically abnormal cases, it is recommended to include cytogenetics (and, for example, the DNA index) in the prognostic armamentarium....

  12. Encodable multiple-fluorescence CdTe@carbon nanoparticles from nanocrystal/colloidal crystal guest-host ensembles.

    Science.gov (United States)

    Guo, Xin; Wang, Cai-Feng; Mao, Li-Hua; Zhang, Jing; Yu, Zi-Yi; Chen, Su

    2013-04-05

    We report herein the controllable generation of encodable multi-fluorescence CdTe@carbon nanoparticles (CdTe@C NPs) via the pyrolysis of quantum dot/photonic crystal (QD/PC) guest-host ensembles. The precursors of CdTe/poly(styrene-co-glycidylmethacrylate) (PS-co-PGMA) QD/PC guest-host ensembles were initially formed via the assembly of epoxy groups of PCs and carboxyl groups on the surface of CdTe QDs, followed by a pyrolysis process to generate CdTe@C NPs. The as-prepared CdTe@C NPs not only integrate the optical properties for both the carbon and CdTe QD constituents, but also enable an impressive enhancement of the fluorescence lifetime for CdTe QDs. The multifarious fluorescent spectra coding for CdTe@C NPs was further generated through regulating the embedded sizes or concentrations of CdTe QDs and the excitation wavelength, and their applications in DNA detection and luminescent patterns were achieved.

  13. A comparison of the intensity contours of fringes due to multiple reflection and spatial hole burning in semiclassical theory of laser

    Indian Academy of Sciences (India)

    Rita Moni Borah; G D Baruah

    2000-02-01

    A comparison between spatial burning in the semiclassical theory of laser and the intensity contours of the fringes due to multiple reflections in a Fabry–Perot cavity is presented. The concept of spatial hole burning is also used in a quantum well system.

  14. Stoke's and anti-Stoke's characteristics of anaerobic and aerobic bacterias at excitation of fluorescence by low-intensity red light: I. Research of anaerobic bacterias

    Science.gov (United States)

    Masychev, Victor I.; Alexandrov, Michail T.

    2000-04-01

    Biopsy or photo dynamic therapy of tumors are usually investigated by fluorescent diagnostics methods. Information on modified method of fluorescence diagnostics of inflammatory diseases is represented in this research. Anaerobic micro organisms are often the cause of these pathological processes. These micro organisms also accompany disbiotic processes in intestines.

  15. Purification of recombinant green fluorescent protein using chromatofocusing with a pH gradient composed of multiple stepwise fronts.

    Science.gov (United States)

    Narahari, C R; Randers-Eichhorn, L; Strong, J C; Ramasubramanyan, N; Rao, G; Frey, D D

    2001-01-01

    Green fluorescent protein (GFP), which fluoresces in the green region of the visible spectrum and is widely used as a reporter for gene expression and regulation, was overexpressed in the JM105 strain of Escherichia coli transformed with pBAD-GFP. A two-step chromatofocusing procedure was used to purify GFP starting from cell lysate, with each step employing a pH gradient extending from pH 5.5 to 4.0. The first chromatofocusing step was performed using a low-pressure column in which a retained stepwise pH front formed by adsorbed buffering species was used to capture GFP directly from clarified cell lysate and selectively focus it into a chromatographic band. The second step utilized a high-performance column under mass overloaded conditions where a similar pH front acted as a protein displacer and led to the formation of a highly concentrated rectangular band of GFP. The overall procedure yielded a 50-fold increase in purity, a 20-fold volume reduction, and a recovery and purity for GFP of 60% and 80%, respectively. Because the method employs a strong-base ion-exchange column packing and low-cost buffers formed with formic and acetic acids instead of the proprietary column packings and polyampholyte elution buffers more generally used for chromatofocusing, it appears to be a practical alternative for the preparative ion-exchange chromatography of GFP in particular and for the recovery of recombinant proteins from cell lysate in general. A discussion is also given concerning the choice of appropriate buffers for the rational design of pH gradients involving retained, stepwise pH fronts that span a given pH range and of the use of the fluorescence properties of GFP for flow visualization and chromatographic process development.

  16. Multiple effects of cadmium on the photosynthetic apparatus of Avicennia germinans L. as probed by OJIP chlorophyll fluorescence measurements

    Energy Technology Data Exchange (ETDEWEB)

    Gonzales-Mendoza, D.; Zapata-Perez, O. [Cinvestav Unidad Merida, Yucatan (Mexico). Dept. de Recursos del Mar; Espadas y Gil, F.; Santamaria, J.M. [Unidad de Biotecnologia, CICY, Yucatan (Mexico)

    2007-03-15

    The toxic effects of cadmium on the photosynthetic apparatus of Avicennia germinans were evaluated by means of the chlorophyll fluorescence transient O-J-I-P. The chlorophyll fluorescence transients were recorded in vivo with high time resolution and analyzed according to the OJIP-test that can quantify the performance of photosystem II. Cadmium-treated plants showed a decrease in yield for primary photochemistry, TR{sup 0}/ABS. The performance index of photosystem II (PSII), PI{sub ABS}, decreased due to cadmium treatment. This performance index is the combination of the indexes of three independent parameters: (1) total number of active reaction centers per absorption (RC/ABS), (2) yield of primary photochemistry (TR{sup 0}/ABS), and (3) efficiency with which a trapped exciton can move an electron into the electron transport chain (ET{sup 0}/TR{sup 0}). Additionally, the F{sub 0}/F{sub v} registered the highest sensitivity to the metal, thus indicating that the water-splitting apparatus of the oxidizing side of PSII is the primary site of action of cadmium. In summary, cadmium affects several targets of photosystem II. More specifically the main targets of cadmium, according to the OJIP-test, can be listed as a decrease in the number of active reaction centers and damage to the activity of the water-splitting complex. (orig.)

  17. Single Molecule Spectroscopy of Fluorescent Proteins

    NARCIS (Netherlands)

    Blum, Christian; Subramaniam, Vinod

    2009-01-01

    The discovery and use of fluorescent proteins has revolutionized cellular biology. Despite the widespread use of visible fluorescent proteins as reporters and sensors in cellular environments the versatile photophysics of fluorescent proteins is still subject to intense research. Understanding the

  18. MO-F-CAMPUS-T-02: Optimizing Orientations of Hundreds of Intensity-Modulated Beams to Treat Multiple Brain Targets

    Energy Technology Data Exchange (ETDEWEB)

    Ma, L; Dong, P; Larson, D [University of California San Francisco, San Francisco, CA (United States); Keeling, V; Hossain, S; Ahmad, S [University of Oklahoma Health Science Center, Oklahoma City, OK (United States); Sahgal, A [University of Toronto, Toronto, ON (Canada)

    2015-06-15

    Purpose: To investigate a new modulated beam orientation optimization (MBOO) approach maximizing treatment planning quality for the state-of-the-art flattening filter free (FFF) beam that has enabled rapid treatments of multiple brain targets. Methods: MBOO selects and optimizes a large number of intensity-modulated beams (400 or more) from all accessible beam angles surrounding a patient’s skull. The optimization algorithm was implemented on a standalone system that interfaced with the 3D Dicom images and structure sets. A standard published data set that consisted of 1 to 12 metastatic brain tumor combinations was selected for MBOO planning. The planning results from various coplanar and non-coplanar configurations via MBOO were then compared with the results obtained from a clinical volume modulated arc therapy (VMAT) delivery system (Truebeam RapidArc, Varian Oncology). Results: When planning a few number of targets (n<4), MBOO produced results equivalent to non-coplanar multi-arc VMAT planning in terms of target volume coverage and normal tissue sparing. For example, the 12-Gy and 4-Gy normal brain volumes for the 3-target plans differed by less than 1 mL ( 3.0 mLvs 3.8 mL; and 35.2 mL vs 36.3 mL, respectively) for MBOO versus VMAT. However, when planning a larger number of targets (n≥4), MBOO significantly reduced the dose to the normal brain as compared to VMAT, though the target volume coverage was equivalent. For example, the 12-Gy and 4-Gy normal brain volumes for the 12-target plans were 10.8 mL vs. 18.0 mL and 217.9 mL vs. 390.0 mL, respectively for the non-coplanar MBOO versus the non-coplanar VMAT treatment plans, yielding a reduction in volume of more than 60% for the case. Conclusion: MBOO is a unique approach for maximizing normal tissue sparing when treating a large number (n≥4) of brain tumors with FFF linear accelerators. Dr Ma and Dr Sahgal are currently on the board of international society of stereotactic radiosurgery. Dr Sahgal has

  19. Increased fluorescence intensity in CaTiO3:Pr3+ phosphor due to NH3 treatment and Nb Co-doping

    Science.gov (United States)

    Holliday, K. S.; Kohlgruber, T. A.; Tran, I. C.; Åberg, D.; Seeley, Z. M.; Bagge-Hansen, M.; Srivastava, A. M.; Cherepy, N. J.; Payne, S. A.

    2016-10-01

    Development of next generation red phosphors for commercial lighting requires understanding of how increased luminescence is achieved by various treatment strategies. In this work, we compare co-doping with Nb to NH3 treatment of CaTiO3:Pr phosphors to reveal a general mechanism responsible for the increased luminescence. The phosphors were synthesized using standard solid-state synthesis techniques and the fluorescence was characterized for potential use in fluorescent lighting, with 254 nm excitation. The lifetime of the fluorescence was determined and used to identify a change in a trap state by the co-doping of Nb5+ in the phosphor. The oxidation state of the Pr was probed by NEXAFS and revealed that both Nb5+ co-doping and NH3 treatment reduced the number of non-fluorescing Pr4+ centers. Calculations were performed to determine the energetically favorable defects. Vacuum annealing was also used to further probe the nature of the trap state. It was determined that NH3 treatments reduce the number of Pr4+ non-fluorescing centers, while Nb5+ co-doping additionally reduces the number of excess oxygen trap states that quench the fluorescence.

  20. Resolving colocalization of bacteria and metal(loid)s on plant root surfaces by combining fluorescence in situ hybridization (FISH) with multiple-energy micro-focused X-ray fluorescence (ME μXRF).

    Science.gov (United States)

    Honeker, Linnea K; Root, Robert A; Chorover, Jon; Maier, Raina M

    2016-12-01

    Metal(loid)-contamination of the environment due to anthropogenic activities is a global problem. Understanding the fate of contaminants requires elucidation of biotic and abiotic factors that influence metal(loid) speciation from molecular to field scales. Improved methods are needed to assess micro-scale processes, such as those occurring at biogeochemical interfaces between plant tissues, microbial cells, and metal(loid)s. Here we present an advanced method that combines fluorescence in situ hybridization (FISH) with synchrotron-based multiple-energy micro-focused X-ray fluorescence microprobe imaging (ME μXRF) to examine colocalization of bacteria and metal(loid)s on root surfaces of plants used to phytostabilize metalliferous mine tailings. Bacteria were visualized on a small root section using SytoBC nucleic acid stain and FISH probes targeting the domain Bacteria and a specific group (Alphaproteobacteria, Gammaproteobacteria, or Actinobacteria). The same root region was then analyzed for elemental distribution and metal(loid) speciation of As and Fe using ME μXRF. The FISH and ME μXRF images were aligned using ImageJ software to correlate microbiological and geochemical results. Results from quantitative analysis of colocalization show a significantly higher fraction of As colocalized with Fe-oxide plaques on the root surfaces (fraction of overlap 0.49±0.19) than to bacteria (0.072±0.052) (pbacteria that colocalized with metal(loid)s, Actinobacteria, known for their metal tolerance, had a higher correlation with both As and Fe than Alphaproteobacteria or Gammaproteobacteria. This method demonstrates how coupling these micro-techniques can expand our understanding of micro-scale interactions between roots, metal(loid)s and microbes, information that should lead to improved mechanistic models of metal(loid) speciation and fate. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Fluorescent dendritic organogels based on 2-(2'-hydroxyphenyl)benzoxazole: emission enhancement and multiple stimuli-responsive properties.

    Science.gov (United States)

    Chen, Hui; Feng, Yu; Deng, Guo-Jun; Liu, Zhi-Xiong; He, Yan-Mei; Fan, Qing-Hua

    2015-07-27

    A new highly efficient and versatile poly(benzyl ether) dendritic organogelator HPB-G1 with 2-(2'-hydroxyphenyl)benzoxazole (HPB) at the focal point has been designed and synthesized. HPB-G1 can form stable organogels toward various apolar and polar organic solvents. Further studies revealed that intermolecular multiple π-π stacking interactions are the main driving forces for the formation of the organogels. Notably, dendron HPB-G1 exhibited a significantly enhanced emission in the gel state in contrast to weak emission in solution. Most interestingly, these dendritic organogels exhibited multiple stimuli-responsive behaviors upon exposure to environmental stimuli, including temperature, sonication, shear stress, and the presence of anions, metal cations, acids/bases, thus leading to reversible sol-gel phase transitions.

  2. Increased signal intensities in the dentate nucleus and globus pallidus on unenhanced T1-weighted images: evidence in children undergoing multiple gadolinium MRI exams

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Houchun H.; Pokorney, Amber; Towbin, Richard B.; Miller, Jeffrey H. [Phoenix Children' s Hospital, Department of Medical Imaging and Radiology, Phoenix, AZ (United States)

    2016-10-15

    Recent reports have suggested residual gadolinium deposition in the brain in subjects undergoing multiple contrast-enhanced MRI exams. These findings have raised some concerns regarding gadolinium-based contrast agent (GBCA) usage and retention in brain tissues. To summarize findings of hyperintense brain structures on precontrast T1-weighted images in 21 children undergoing multiple GBCA MRI exams. This retrospective study involved 21 patients, each of whom received multiple MRI examinations (range: 5-37 exams) with GBCA over the course of their medical treatment (duration from first to most recent exam: 1.2-12.9 years). The patients were between 0.9 and 14.4 years of age at the time of their first GBCA exam. Regions of interest were drawn in the dentate nucleus and the globus pallidus on 2-D fast spin echo images acquired at 1.5 T. The signal intensities of these two structures were normalized by that of the corpus callosum genu. Signal intensity ratios from these patients were compared to control patients of similar ages who have never received GBCA. Signal intensity ratios increased between the first and the most recent MRI exam in all 21 patients receiving GBCA, with an increase of 18.6%±12.7% (range: 0.5% to 47.5%) for the dentate nucleus and 12.4%±7.4% (range: -1.2% to 33.7%) for the globus pallidus (P<0.0001). Signal intensity ratios were also higher in GBCA patients than in controls (P<0.01). The degree of signal intensity enhancement did not correlate with statistical significance to the cumulative number or volume of GBCA administrations each patient received, the patient's age or the elapsed time between the first and most recent GBCA MRI exams. These results in children are consistent with recent findings in adults, suggesting possible gadolinium deposition in the brain. (orig.)

  3. Fluorescence antibunching microscopy

    CERN Document Server

    Schwartz, Osip

    2011-01-01

    Breaking the diffraction limit in microscopy by utilizing quantum properties of light has been the goal of intense research in the recent years. We propose a quantum superresolution technique based on non-classical emission statistics of fluorescent markers, routinely used as contrast labels for bio-imaging. The technique can be readily implemented using standard fluorescence microscopy equipment.

  4. Fluorescence of atopic allergens

    NARCIS (Netherlands)

    Berrens, L.

    1967-01-01

    Purified atopic allergens have been found to emit flue fluorescence upon irradiation with ultraviolet light of 365 mμ wavelength. The maximum of fluorescence is in the region 445–490 mμ and the intensity is of the same order of magnitude for different atopic allergens. Synthetic model compounds, inc

  5. Development of indirect competitive fluorescence immunoassay for 2,2′,4,4′-tetrabromodiphenyl ether using DNA/dye conjugate as antibody multiple labels

    Institute of Scientific and Technical Information of China (English)

    Zi-Yan Fan; Young Soo Keum; Qing-Xiao Li; Weilin L. Shelver; Liang-Hong Guo

    2012-01-01

    An indirect competitive fluorescence immunoassay using a DNA/dye conjugate as antibody multiple labels was developed on 96-well plates for the identification and quantification of 2,2′,4,4′-tetrabromodiphenyl ether (BDE-47) in aqueous samples.A hapten,2,4,2′-tribromodiphenyl ether-4′-aldehyde,was synthesized,and was conjugated to bovine serum albumin to form a coating antigen,Specific recognition of the antigen by anti-PBDE antiserum was confirmed by a surface plasmon resonance measurement.In the immunoassay,the coating antigen was adsorbed on a 96-well plate first,and a sample,antiserum and biotinylated goat anti-rabbit secondary antibody were then added and reacted sequentially.A biotinylated,double-stranded DNA with 219 base pairs was attached to the secondary antibody by using streptavidin as a molecular bridge.In situ multiple labeling of the antibody was accomplished after addition of a DNA-binding fluorescent dye,SYBR Green I.The working range of the immunoassay for the BDE-47 standard was 3.1-390 μg/L,with an IC50 value of 15.6 μg/L.The calculated LOD of the immunoassay is 0.73 μg/L.The immunoassay demonstrated relatively high selectivity for BDE-47,showing very low cross-reactivity (< 3%) with BDE-15,BDE-153 and BDE-209.With a spiked river water sample containing 50 μg/L BDE-47,quantification by the immunoassay was 41.9 μg/L,which compared well with the standard GC-ECD method (45.7 μg/L).The developed immunoassay provides a rapid screening tool for polybrominated diphenyl ethers in environmental samples.

  6. The influence of electron multiplication and internal X-ray fluorescence on the performance of a scintillator-based gamma camera

    Energy Technology Data Exchange (ETDEWEB)

    Hall, David J., E-mail: d.j.hall@open.ac.uk [e2v centre for electronic imaging, The Open University, Walton Hall, Milton Keynes MK7 6AA (United Kingdom); Holland, Andrew; Soman, Matthew [e2v centre for electronic imaging, The Open University, Walton Hall, Milton Keynes MK7 6AA (United Kingdom)

    2012-06-21

    When considering the 'standard' gamma-camera, one might picture an array of photo-multiplier tubes or a similar array of small-area detectors. This array of imaging detectors would be attached to a corresponding array of scintillator modules (or a solid layer of scintillator) in order to give a high detection efficiency in the energy region of interest, usually 8-140 keV. Over recent years, developments of gamma-cameras capable of achieving much higher spatial resolutions have led to a new range of systems based on Charge-Coupled Devices with some form of signal multiplication between the scintillator and the CCD in order for one to distinguish the light output from the scintillator above the CCD noise. The use of an Electron-Multiplying Charge-Coupled Device (EM-CCD) incorporates the gain process within the CCD through a form of 'impact ionisation', however, the gain process introduces an 'excess noise factor' due to the probabilistic nature of impact ionisation and this additional noise consequently has an impact on the spatial and spectral resolution of the detector. Internal fluorescence in the scintillator, producing K-shell X-ray fluorescence photons that can be detected alongside the incident gamma-rays, also has a major impact on the imaging capabilities of gamma-cameras. This impact varies dramatically from the low spatial resolution to high spatial resolution camera system. Through a process of simulation and experimental testing focussed on the high spatial resolution (EM-CCD based) variant, the factors affecting the performance of gamma-camera systems are discussed and the results lead to important conclusions to be considered for the development of future systems. This paper presents a study into the influence of the EM-CCD gain process and the internal X-ray fluorescence in the scintillator on the performance of scintillator-based gamma cameras (CCD-based or otherwise), making use of Monte Carlo simulations to demonstrate

  7. Cancer detection by quantitative fluorescence image analysis.

    Science.gov (United States)

    Parry, W L; Hemstreet, G P

    1988-02-01

    Quantitative fluorescence image analysis is a rapidly evolving biophysical cytochemical technology with the potential for multiple clinical and basic research applications. We report the application of this technique for bladder cancer detection and discuss its potential usefulness as an adjunct to methods used currently by urologists for the diagnosis and management of bladder cancer. Quantitative fluorescence image analysis is a cytological method that incorporates 2 diagnostic techniques, quantitation of nuclear deoxyribonucleic acid and morphometric analysis, in a single semiautomated system to facilitate the identification of rare events, that is individual cancer cells. When compared to routine cytopathology for detection of bladder cancer in symptomatic patients, quantitative fluorescence image analysis demonstrated greater sensitivity (76 versus 33 per cent) for the detection of low grade transitional cell carcinoma. The specificity of quantitative fluorescence image analysis in a small control group was 94 per cent and with the manual method for quantitation of absolute nuclear fluorescence intensity in the screening of high risk asymptomatic subjects the specificity was 96.7 per cent. The more familiar flow cytometry is another fluorescence technique for measurement of nuclear deoxyribonucleic acid. However, rather than identifying individual cancer cells, flow cytometry identifies cellular pattern distributions, that is the ratio of normal to abnormal cells. Numerous studies by others have shown that flow cytometry is a sensitive method to monitor patients with diagnosed urological disease. Based upon results in separate quantitative fluorescence image analysis and flow cytometry studies, it appears that these 2 fluorescence techniques may be complementary tools for urological screening, diagnosis and management, and that they also may be useful separately or in combination to elucidate the oncogenic process, determine the biological potential of tumors

  8. Multiple Linear Regression Analysis Indicates Association of P-Glycoprotein Substrate or Inhibitor Character with Bitterness Intensity, Measured with a Sensor.

    Science.gov (United States)

    Yano, Kentaro; Mita, Suzune; Morimoto, Kaori; Haraguchi, Tamami; Arakawa, Hiroshi; Yoshida, Miyako; Yamashita, Fumiyoshi; Uchida, Takahiro; Ogihara, Takuo

    2015-09-01

    P-glycoprotein (P-gp) regulates absorption of many drugs in the gastrointestinal tract and their accumulation in tumor tissues, but the basis of substrate recognition by P-gp remains unclear. Bitter-tasting phenylthiocarbamide, which stimulates taste receptor 2 member 38 (T2R38), increases P-gp activity and is a substrate of P-gp. This led us to hypothesize that bitterness intensity might be a predictor of P-gp-inhibitor/substrate status. Here, we measured the bitterness intensity of a panel of P-gp substrates and nonsubstrates with various taste sensors, and used multiple linear regression analysis to examine the relationship between P-gp-inhibitor/substrate status and various physical properties, including intensity of bitter taste measured with the taste sensor. We calculated the first principal component analysis score (PC1) as the representative value of bitterness, as all taste sensor's outputs shared significant correlation. The P-gp substrates showed remarkably greater mean bitterness intensity than non-P-gp substrates. We found that Km value of P-gp substrates were correlated with molecular weight, log P, and PC1 value, and the coefficient of determination (R(2) ) of the linear regression equation was 0.63. This relationship might be useful as an aid to predict P-gp substrate status at an early stage of drug discovery.

  9. Multiple ionization of atom clusters by intense soft X-rays from a free-electron laser.

    Science.gov (United States)

    Wabnitz, H; Bittner, L; de Castro, A R B; Döhrmann, R; Gürtler, P; Laarmann, T; Laasch, W; Schulz, J; Swiderski, A; von Haeften, K; Möller, T; Faatz, B; Fateev, A; Feldhaus, J; Gerth, C; Hahn, U; Saldin, E; Schneidmiller, E; Sytchev, K; Tiedtke, K; Treusch, R; Yurkov, M

    2002-12-05

    Intense radiation from lasers has opened up many new areas of research in physics and chemistry, and has revolutionized optical technology. So far, most work in the field of nonlinear processes has been restricted to infrared, visible and ultraviolet light, although progress in the development of X-ray lasers has been made recently. With the advent of a free-electron laser in the soft-X-ray regime below 100 nm wavelength, a new light source is now available for experiments with intense, short-wavelength radiation that could be used to obtain deeper insights into the structure of matter. Other free-electron sources with even shorter wavelengths are planned for the future. Here we present initial results from a study of the interaction of soft X-ray radiation, generated by a free-electron laser, with Xe atoms and clusters. We find that, whereas Xe atoms become only singly ionized by the absorption of single photons, absorption in clusters is strongly enhanced. On average, each atom in large clusters absorbs up to 400 eV, corresponding to 30 photons. We suggest that the clusters are heated up and electrons are emitted after acquiring sufficient energy. The clusters finally disintegrate completely by Coulomb explosion.

  10. L X-ray energy shifts and intensity ratios in tantalum with C and N ions – multiple vacancies in M, N and O shells

    Indian Academy of Sciences (India)

    Y Ramakrishna; K Ramachandra Rao; G J Naga Raju; K Bhaskara Rao; V Seshagiri Rao; P Venkateswarlu; S Bhuloka Reddy

    2002-10-01

    The energy shifts and intensity ratios of different L X-ray components in tantalum element due to 10 MeV carbon and 12 MeV nitrogen ions are estimated. From the observed energy shifts, the possible number of simultaneous vacancies in M shell are estimated. A comparison of L/L 2,15, L 1/L 1 and L 2,3/L 4,4 with the ratios due to Scofield theoretical transition rates indicate that the number of multiple vacancies in N shell are higher than the vacancies in M and O shell. Employing Larkin’s statistical scaling procedure, the number of possible multiple vacancies in N and O shells are estimated quantitatively.

  11. The detectability of nitrous oxide mitigation efficacy in intensively grazed pastures using a multiple-plot micrometeorological technique

    Directory of Open Access Journals (Sweden)

    A. M. S. McMillan

    2014-05-01

    Full Text Available Methodologies are required to verify agricultural greenhouse gas mitigation at scales relevant to farm management. Micrometeorological techniques provide a viable approach for comparing fluxes between fields receiving mitigation treatments and control fields. However, they have rarely been applied to spatially verifying treatments aimed at mitigating nitrous oxide emission from intensively grazed pastoral systems. We deployed a micrometeorological system to compare N2O flux among several ~1.5 ha plots in intensively grazed dairy pasture. The sample collection and measurement system is referred to as the Field-Scale Nitrous Oxide Mitigation Assessment System (FS-NOMAS and used a tuneable diode laser absorption spectrometer to measure N2O gradients to high precision at four locations along a 300 m transect. The utility of the FS-NOMAS to assess mitigation efficacy depends largely on its ability to resolve very small vertical N2O gradients. The performance of the FS-NOMAS was assessed in this respect in laboratory and field-based studies. The FS-NOMAS could reliably resolve gradients of 0.039 ppb between a height of 0.5 and 1.0 m. The gradient resolution achieved corresponded to the ability to detect an inter-plot N2O flux difference of 26 μg N2O–N m−2 h−1 under the most commonly encountered conditions of atmospheric mixing (quantified here by a turbulent transfer coefficient, but this ranged from 11 to 59 μg N2O–N m−2 h−1 as the transfer coefficient ranged between its 5th and 95th percentile. Assuming a likely value of 100 μg N2O–N m−2 h−1 for post-grazing N2O fluxes from intensively grazed New Zealand dairy pasture, the system described here would be capable of detecting a mitigation efficacy of 26% for a single (40 min comparison. We demonstrate that the system has considerably greater sensitivity to treatment effects by measuring cumulative fluxes over extended periods.

  12. Polar plot representation of time-resolved fluorescence.

    Science.gov (United States)

    Eichorst, John Paul; Wen Teng, Kai; Clegg, Robert M

    2014-01-01

    Measuring changes in a molecule's fluorescence emission is a common technique to study complex biological systems such as cells and tissues. Although the steady-state fluorescence intensity is frequently used, measuring the average amount of time that a molecule spends in the excited state (the fluorescence lifetime) reveals more detailed information about its local environment. The lifetime is measured in the time domain by detecting directly the decay of fluorescence following excitation by short pulse of light. The lifetime can also be measured in the frequency domain by recording the phase and amplitude of oscillation in the emitted fluorescence of the sample in response to repetitively modulated excitation light. In either the time or frequency domain, the analysis of data to extract lifetimes can be computationally intensive. For example, a variety of iterative fitting algorithms already exist to determine lifetimes from samples that contain multiple fluorescing species. However, recently a method of analysis referred to as the polar plot (or phasor plot) is a graphical tool that projects the time-dependent features of the sample's fluorescence in either the time or frequency domain into the Cartesian plane to characterize the sample's lifetime. The coordinate transformations of the polar plot require only the raw data, and hence, there are no uncertainties from extensive corrections or time-consuming fitting in this analysis. In this chapter, the history and mathematical background of the polar plot will be presented along with examples that highlight how it can be used in both cuvette-based and imaging applications.

  13. Fluorescent multiple staining and CASA system to assess boar sperm viability and membranes integrity in short and long-term extenders.

    Science.gov (United States)

    Lange-Consiglio, A; Meucci, A; Cremonesi, F

    2013-01-01

    The aim of this study was to assess the effect on boar spermatozoa quality of in vitro storage in short and long-term extenders by fluorescent multiple staining (FMS) and computer assisted semen analyzer (CASA). Fresh ejaculates from three healthy, sexually mature boars were diluted with equal volumes of six short-term or three long-term commercial extenders and stored at 19°C for 6 days (short-term) or 12 days (long-term). The integrity of spermatozoa membranes was analyzed by FMS using propidium iodide, 5,5',6,6'-tetrachloro-1,1',3,3' tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) and fluorescein isothiocyanate-conjugated peanut agglutinin (PNA). The results obtained from this staining were compared with spermatozoa motility assessed by CASA. Our study showed that the number of viable spermatozoa with non-reacted acrosomes and intact mitochondria was positively correlated with the rate of motile spermatozoa (r(2)>0.9) irrespective of the extender used. In all extenders the number of motile spermatozoa significantly decreased as preservation period increased (P<0.05). FMS test is a potent indicator of sperm motility because it analyses mitochondrial integrity independently from observable alterations in motility. The best performing extenders were BTS for short-term storage and TRI-x-Cell for long-term storage.

  14. Fluorescent multiple staining and CASA system to assess boar sperm viability and membranes integrity in short and long-term extenders

    Directory of Open Access Journals (Sweden)

    F. Cremonesi

    2013-03-01

    Full Text Available The aim of this study was to assess the effect on boar spermatozoa quality of in vitro storage in short and long-term extenders by fluorescent multiple staining (FMS and computer assisted semen analyzer (CASA. Fresh ejaculates from three healthy, sexually mature boars were diluted with equal volumes of six short-term or three long-term commercial extenders and stored at 19°C for 6 days (short-term or 12 days (long-term. The integrity of spermatozoa membranes was analyzed by FMS using propidium iodide, 5,5’,6,6’-tetrachloro-1,1’,3,3’ tetraethylbenzimidazolyl-carbocyanine iodide (JC-1 and fluorescein isothiocyanate-conjugated peanut agglutinin (PNA. The results obtained from this staining were compared with spermatozoa motility assessed by CASA. Our study showed that the number of viable spermatozoa with non-reacted acrosomes and intact mitochondria was positively correlated with the rate of motile spermatozoa (r2>0.9 irrespective of the extender used. In all extenders the number of motile spermatozoa significantly decreased as preservation period increased (P<0.05. FMS test is a potent indicator of sperm motility because it analyses mitochondrial integrity independently from observable alterations in motility. The best performing extenders were BTS for short-term storage and TRI-x-Cell for long-term storage.

  15. Reactive Astrocytes Expressing Intense Estrogen Receptor-alpha Immunoreactivities Have Much Elongated Cytoplasmic Processes: An Autopsy Case of Human Cerebellar Tissue with Multiple Genitourinary and Gastrointestinal Anomalies

    Science.gov (United States)

    Kim, Eo-Jin; Oh, Chang Seok; Kim, Jaehyup; Kim, Wu Ho; Chung, Yoon Hee

    2007-01-01

    We performed an immunohistochemical study on the estrogen receptor alpha (ER-α) distribution in the cerebellum of a human neonate with multiple congenital anomalies, that had been acquired during autopsy. Although the exact pathology in the brain was not clearly elucidated in this study, an unidentified stressful condition might have induced the astrocytes into reactive states. In this immunohistochemical study on the neonatal cerebellum with multiple congenital anomalies, intense ER-α immunoreactivities (IRs) were localized mainly within the white matter even though ER-α IRs were known to be mainly localized in neurons. Double immunohistochemical staining showed that ER-α IR cells were reactive astrocytes, but not neurons. Interestingly, there were differences in the process length among the reactive astrocytes showing ER-α IRs. Our quantitative data confirmed that among the glial fibrillary acidic protein (GFAP)-expressing reactive astrocytes, the cells exhibiting intense ER-α IRs have much longer cytoplasmic processes and relatively weaker GFAP IRs. Taken together, the elongated processes of reactive astrocytes might be due to decreased expression of GFAP, which might be induced by elevated expression of ER-α even though the elucidation of the exact mechanism needs further studies. PMID:17982251

  16. Biomolecule-to-fluorescent-color encoder: modulation of fluorescence emission via DNA structural changes.

    Science.gov (United States)

    Nishimura, Takahiro; Ogura, Yusuke; Yamada, Kenji; Ohno, Yuko; Tanida, Jun

    2014-07-01

    A biomolecule-to-fluorescent-color (B/F) encoder for optical readout of biomolecular information is proposed. In the B/F encoder, a set of fluorescence wavelengths and their intensity levels are used for coding of a biomolecular signal. A hybridization chain reaction of hairpin DNAs labeled with fluorescent reporters was performed to generate the fluorescence color codes. The fluorescence is modulated via fluorescence resonance energy transfer, which is controlled by DNA structural changes. The results demonstrate that fluorescent color codes can be configured based on two wavelengths and five intensities using the B/F encoder, and the assigned codes can be retrieved via fluorescence measurements.

  17. Quantification of fluorescent reporters in plant cells.

    Science.gov (United States)

    Pound, Michael; French, Andrew P; Wells, Darren M

    2015-01-01

    Fluorescent reporters are powerful tools for plant research. Many studies require accurate determination of fluorescence intensity and localization. Here, we describe protocols for the quantification of fluorescence intensity in plant cells from confocal laser scanning microscope images using semiautomated software and image analysis techniques.

  18. The paradox between low shock-stage and evidence for compaction in CM carbonaceous chondrites explained by multiple low-intensity impacts

    Science.gov (United States)

    Lindgren, Paula; Hanna, Romy D.; Dobson, Katherine J.; Tomkinson, Tim; Lee, Martin R.

    2015-01-01

    Petrographic analysis of eight CM carbonaceous chondrites (EET 96029, LAP 031166, LON 94101, MET 01072, Murchison, Murray, SCO 06043, QUE 93005) by electron imaging and diffraction, and X-ray computed tomography, reveals that six of them have a petrofabric defined by shock flattened chondrules. With the exception of Murchison, those CMs that have a strong petrofabric also contain open or mineralized fractures, indicating that tensional stresses accompanying the impacts were sufficient to locally exceed the yield strength of the meteorite matrix. The CMs studied span a wide range of petrologic subtypes, and in common with Rubin (2012) we find that the strength of their petrofabrics increases with their degree of aqueous alteration. This correspondence suggests that impacts were responsible for enhancing alteration, probably because the fracture networks they formed tapped fluid reservoirs elsewhere in the parent body. Two meteorites that do not fit this pattern are MET 01072 and Murchison; both have a strong petrofabric but are relatively unaltered. In the case of MET 01072, impact deformation is likely to have postdated parent body aqueous activity. The same may also be true for Murchison, but as this meteorite also lacks fractures and veins, its chondrules were most likely flattened by multiple low intensity impacts. Multiphase deformation of Murchison is also revealed by the microstructures of calcite grains, and chondrule-defined petrofabrics as revealed by X-ray computed tomography. The contradiction between the commonplace evidence for impact-deformation of CMs and their low shock stages (most belong to S1) can be explained by most if not all having been exposed to multiple low intensity (i.e., enhanced by those impacts that were of sufficient intensity to open high permeability fracture networks that could connect to fluid reservoirs.

  19. A novel approach to identify non-palpable breast lesions combining fluorescent liposomes and magnetic resonance-guided high intensity focused ultrasound-triggered release

    NARCIS (Netherlands)

    Oerlemans, Chris; Nijsen, Frank; van Amersfoort, Miranda; van Bloois, Louis; Heijman, Edwin; Luijten, Peter; Mali, Willem; Storm, Gert

    2011-01-01

    The combination of fluorescein-containing liposomes (FCL) and magnetic resonance-guided high intensity focused ultrasound (MR-HIFU)-triggered release is a promising approach for lesion demarcation and more efficient removal of non-palpable breast lesions. Exposure of FCL to ablation temperatures (60

  20. Computed Flow and Fluorescence Over the Ocular Surface

    CERN Document Server

    Li, Longfei; Henshaw, W D; King-Smith, P E

    2016-01-01

    Fluorescein is perhaps the most commonly used substance to visualize tear film thickness and dynamics; better understanding of this process aids understanding of dry eye syndrome which afflicts millions of people. We study a mathematical model for tear film flow, evaporation, solutal transport and fluorescence over the exposed ocular surface during the interblink. Transport of the fluorescein ion by fluid flow in the tear film affects the intensity of fluorescence via changes in concentration and tear film thickness. Evaporation causes increased osmolarity and potential irritation over the ocular surface; it also alters fluorescein concentration and thus fluorescence. Using thinning rates from in vivo measurements together with thin film equations for flow and transport of multiple solutes, we compute dynamic results for tear film quantities of interest. We compare our computed intensity distributions with in vivo observations. A number of experimental features are recovered by the model.

  1. Effect of solution pH value changes on fluorescence intensity of magnetic-luminescent Fe3O4@Gd2O3:Eu3+ nanoparticles

    Institute of Scientific and Technical Information of China (English)

    吴拓; 潘桦滟; 陈如标; 罗东; 张宏; 沈晔; 李旸晖; 王乐

    2016-01-01

    In this paper, bifunctional Fe3O4@Gd2O3:Eu3+ core-shell nanoparticles with both magnetic and fluorescent properties were synthesized through a urea homogeneous precipitation (UHP) method. Particular emphasis was placed on investigating the influence of the solution pH value on the photoluminescence of the core-shell nanocomposites. It showed that the samples treated at the solution of pH=3.0 had the highest luminescence due to the enhanced crystallinity and size uniformity of nanoparticles. The Fe3O4@Gd2O3:Eu3+ nanocomposites exhibited an almost spherical shape with a mean diameter of 60 nm, and had strong red emis-sions of Eu3+ at 612 nm as well as good magnetization with the saturation magnetization of 1.29 emu/g. It thus indicated that the core-shell nanocomposites investigated has great potential in biomedical applications.

  2. Intense inflammation and nerve damage in early multiple sclerosis subsides at older age: a reflection by cerebrospinal fluid biomarkers.

    Directory of Open Access Journals (Sweden)

    Mohsen Khademi

    Full Text Available Inflammatory mediators have crucial roles in leukocyte recruitment and subsequent central nervous system (CNS neuroinflammation. The extent of neuronal injury and axonal loss are associated with the degree of CNS inflammation and determine physical disability in multiple sclerosis (MS. The aim of this study was to explore possible associations between a panel of selected cerebrospinal fluid biomarkers and robust clinical and demographic parameters in a large cohort of patients with MS and controls (n = 1066 using data-driven multivariate analysis. Levels of matrix metalloproteinase 9 (MMP9, chemokine (C-X-C motif ligand 13 (CXCL13, osteopontin (OPN and neurofilament-light chain (NFL were measured by ELISA in 548 subjects comprising different MS subtypes (relapsing-remitting, secondary progressive and primary progressive, clinically isolated syndrome and persons with other neurological diseases with or without signs of inflammation/infection. Principal component analyses and orthogonal partial least squares methods were used for unsupervised and supervised interrogation of the data. Models were validated using data from a further 518 subjects in which one or more of the four selected markers were measured. There was a significant association between increased patient age and lower levels of CXCL13, MMP9 and NFL. CXCL13 levels correlated well with MMP9 in the younger age groups, but less so in older patients, and after approximately 54 years of age the levels of CXCL13 and MMP9 were consistently low. CXCL13 and MMP9 levels also correlated well with both NFL and OPN in younger patients. We demonstrate a strong effect of age on both inflammatory and neurodegenerative biomarkers in a large cohort of MS patients. The findings support an early use of adequate immunomodulatory disease modifying drugs, especially in younger patients, and may provide a biological explanation for the relative inefficacy of such treatments in older patients at later

  3. Intense inflammation and nerve damage in early multiple sclerosis subsides at older age: a reflection by cerebrospinal fluid biomarkers.

    Science.gov (United States)

    Khademi, Mohsen; Dring, Ann M; Gilthorpe, Jonathan D; Wuolikainen, Anna; Al Nimer, Faiez; Harris, Robert A; Andersson, Magnus; Brundin, Lou; Piehl, Fredrik; Olsson, Tomas; Svenningsson, Anders

    2013-01-01

    Inflammatory mediators have crucial roles in leukocyte recruitment and subsequent central nervous system (CNS) neuroinflammation. The extent of neuronal injury and axonal loss are associated with the degree of CNS inflammation and determine physical disability in multiple sclerosis (MS). The aim of this study was to explore possible associations between a panel of selected cerebrospinal fluid biomarkers and robust clinical and demographic parameters in a large cohort of patients with MS and controls (n = 1066) using data-driven multivariate analysis. Levels of matrix metalloproteinase 9 (MMP9), chemokine (C-X-C motif) ligand 13 (CXCL13), osteopontin (OPN) and neurofilament-light chain (NFL) were measured by ELISA in 548 subjects comprising different MS subtypes (relapsing-remitting, secondary progressive and primary progressive), clinically isolated syndrome and persons with other neurological diseases with or without signs of inflammation/infection. Principal component analyses and orthogonal partial least squares methods were used for unsupervised and supervised interrogation of the data. Models were validated using data from a further 518 subjects in which one or more of the four selected markers were measured. There was a significant association between increased patient age and lower levels of CXCL13, MMP9 and NFL. CXCL13 levels correlated well with MMP9 in the younger age groups, but less so in older patients, and after approximately 54 years of age the levels of CXCL13 and MMP9 were consistently low. CXCL13 and MMP9 levels also correlated well with both NFL and OPN in younger patients. We demonstrate a strong effect of age on both inflammatory and neurodegenerative biomarkers in a large cohort of MS patients. The findings support an early use of adequate immunomodulatory disease modifying drugs, especially in younger patients, and may provide a biological explanation for the relative inefficacy of such treatments in older patients at later disease stages.

  4. SynPAnal: software for rapid quantification of the density and intensity of protein puncta from fluorescence microscopy images of neurons.

    Directory of Open Access Journals (Sweden)

    Eric Danielson

    Full Text Available Continuous modification of the protein composition at synapses is a driving force for the plastic changes of synaptic strength, and provides the fundamental molecular mechanism of synaptic plasticity and information storage in the brain. Studying synaptic protein turnover is not only important for understanding learning and memory, but also has direct implication for understanding pathological conditions like aging, neurodegenerative diseases, and psychiatric disorders. Proteins involved in synaptic transmission and synaptic plasticity are typically concentrated at synapses of neurons and thus appear as puncta (clusters in immunofluorescence microscopy images. Quantitative measurement of the changes in puncta density, intensity, and sizes of specific proteins provide valuable information on their function in synaptic transmission, circuit development, synaptic plasticity, and synaptopathy. Unfortunately, puncta quantification is very labor intensive and time consuming. In this article, we describe a software tool designed for the rapid semi-automatic detection and quantification of synaptic protein puncta from 2D immunofluorescence images generated by confocal laser scanning microscopy. The software, dubbed as SynPAnal (for Synaptic Puncta Analysis, streamlines data quantification for puncta density and average intensity, thereby increases data analysis throughput compared to a manual method. SynPAnal is stand-alone software written using the JAVA programming language, and thus is portable and platform-free.

  5. Synthesis and Fluorescence Spectra of Triazolylcoumarin Fluorescent Dyes

    Institute of Scientific and Technical Information of China (English)

    PENG Xian-fu; LI Hong-qi

    2009-01-01

    Much attention is devoted to fluorescent dyes especially those with potential in versatile applications. Reactions under "click" conditions between nonfluorescent 3 - azidocoumarins and terminal alkynes produced 3 -(1, 2, 3- triazol- 1 - yl)cournarins, a novel type of fluorescent dyes with intense fluorescence. The structures of the new coumarins were characterized by 1H NMR, MS, and IR spectra. Fluorescence spectra measurement demonstrated excellent fluorescence performance of the triazolylcoumarins and this click reaction is a promising candidate for bioconjugation and bioimaging applications since both azide and alkynes are quite inert to biological systems.

  6. Clinical applicability and prognostic significance of molecular response assessed by fluorescent-PCR of immunoglobulin genes in multiple myeloma. Results from a GEM/PETHEMA study.

    Science.gov (United States)

    Martinez-Lopez, Joaquin; Fernández-Redondo, Elena; García-Sánz, Ramón; Montalbán, María Angeles; Martínez-Sánchez, Pilar; Pavia, Bruno; Mateos, María Victoria; Rosiñol, Laura; Martín, Marisa; Ayala, Rosa; Martínez, Rafael; Blanchard, María Jesus; Alegre, Adrian; Besalduch, Joan; Bargay, Joan; Hernandez, Miguel T; Sarasquete, María Eugenia; Sanchez-Godoy, Pedro; Fernández, Manuela; Blade, Joan; San Miguel, Jesús F; Lahuerta, Juan Jose

    2013-12-01

    Minimal residual disease monitoring is becoming increasingly important in multiple myeloma (MM), but multiparameter flow cytometry (MFC) and allele-specific oligonucleotide polymerase chain reaction (ASO-PCR) techniques are not routinely available. This study investigated the prognostic influence of achieving molecular response assessed by fluorescent-PCR (F-PCR) in 130 newly diagnosed MM patients from Grupo Español Multidisciplinar de Melanoma (GEM)2000/GEM05 trials (NCT00560053, NCT00443235, NCT00464217) who achieved almost very good partial response after induction therapy. As a reference, we used the results observed with simultaneous MFC. F-PCR at diagnosis was performed on DNA using three different multiplex PCRs: IGH D-J, IGK V-J and KDE rearrangements. The applicability of F-PCR was 91·5%. After induction therapy, 64 patients achieved molecular response and 66 non-molecular response; median progression-free survival (PFS) was 61 versus 36 months, respectively (P = 0·001). Median overall survival (OS) was not reached (NR) in molecular response patients (5-year survival: 75%) versus 66 months in the non-molecular response group (P = 0·03). The corresponding PFS and OS values for patients with immunophenotypic versus non-immunophenotypic response were 67 versus 42 months (P = 0·005) and NR (5-year survival: 95%) versus 69 months (P = 0·004), respectively. F-PCR analysis is a rapid, affordable, and easily performable technique that, in some circumstances, may be a valid approach for minimal residual disease investigations in MM.

  7. In situ observation of the growth of biofouling layer in osmotic membrane bioreactors by multiple fluorescence labeling and confocal laser scanning microscopy.

    Science.gov (United States)

    Yuan, Bo; Wang, Xinhua; Tang, Chuyang; Li, Xiufen; Yu, Guanghui

    2015-05-15

    Since the concept of the osmotic membrane bioreactor (OMBR) was introduced in 2008, it has attracted growing interests for its potential applications in wastewater treatment and reclamation; however, the fouling mechanisms of forward osmosis (FO) membrane especially the development of biofouling layer in the OMBR are not yet clear. Here, the fouled FO membranes were obtained from the OMBRs on days 3, 8 and 25 in sequence, and then the structure and growing rule of the biofouling layer formed on the FO membrane samples were in-situ characterized by multiple fluorescence labeling and confocal laser scanning microscopy (CLSM). CLSM images indicated that the variations in abundance and distribution of polysaccharides, proteins and microorganisms in the biofouling layer during the operation of OMBRs were significantly different. Before the 8th day, their biovolume dramatically increased. Subsequently, the biovolumes of β-d-glucopyranose polysaccharides and proteins continued increasing and leveled off after 8 days, respectively, while the biovolumes of α-d-glucopyranose polysaccharides and microorganisms decreased. Extracellular polymeric substances (EPS) played a significant role in the formation and growth of biofouling layer, while the microorganisms were seldom detected on the upper fouling layer after 3 days. Based on the results obtained in this study, the growth of biofouling layer on the FO membrane surface in the OMBR could be divided into three stages. Initially, EPS was firstly deposited on the FO membrane surface, and then microorganisms associated with EPS located in the initial depositing layer to form clusters. After that, the dramatic increase of the clusters of EPS and microorganisms resulted in the quick growth of biofouling layer during the flux decline of the OMBR. However, when the water flux became stable in the OMBR, some microorganisms and EPS would be detached from the FO membrane surface.

  8. Microfluidic flow cytometer for quantifying photobleaching of fluorescent proteins in cells.

    Science.gov (United States)

    Lubbeck, Jennifer L; Dean, Kevin M; Ma, Hairong; Palmer, Amy E; Jimenez, Ralph

    2012-05-01

    Traditional flow cytometers are capable of rapid cellular assays on the basis of fluorescence intensity and light scatter. Microfluidic flow cytometers have largely followed the same path of technological development as their traditional counterparts; however, the significantly smaller transport distance and resulting lower cell speeds in microchannels provides for the opportunity to detect novel spectroscopic signatures based on multiple, nontemporally coincident excitation beams. Here, we characterize the design and operation of a cytometer with a three-beam, probe/bleach/probe geometry, employing HeLa suspension cells expressing fluorescent proteins. The data collection rate exceeds 20 cells/s under a range of beam intensities (5 kW to 179 kW/cm(2)). The measured percent photobleaching (ratio of fluorescence intensities excited by the first and third beams: S(beam3)/S(beam1)) partially resolves a mixture of four red fluorescent proteins in mixed samples. Photokinetic simulations are presented and demonstrate that the percent photobleaching reflects a combination of the reversible and irreversible photobleaching kinetics. By introducing a photobleaching optical signature, which complements traditional fluorescence intensity-based detection, this method adds another dimension to multichannel fluorescence cytometry and provides a means for flow-cytometry-based screening of directed libraries of fluorescent protein photobleaching.

  9. The ROCK inhibitor Y-27632 improves recovery of human embryonic stem cells after fluorescence-activated cell sorting with multiple cell surface markers.

    Directory of Open Access Journals (Sweden)

    Nil Emre

    Full Text Available BACKGROUND: Due to the inherent sensitivity of human embryonic stem cells (hESCs to manipulations, the recovery and survival of hESCs after fluorescence-activated cell sorting (FACS can be low. Additionally, a well characterized and robust methodology for performing FACS on hESCs using multiple-cell surface markers has not been described. The p160-Rho-associated coiled kinase (ROCK inhibitor, Y-27632, previously has been identified as enhancing survival of hESCs upon single-cell dissociation, as well as enhancing recovery from cryopreservation. Here we examined the application of Y-27632 to hESCs after FACS to improve survival in both feeder-dependent and feeder-independent growth conditions. METHODOLOGY/PRINCIPAL FINDINGS: HESCs were sorted using markers for SSEA-3, TRA-1-81, and SSEA-1. Cells were plated after sorting for 24 hours in either the presence or the absence of Y-27632. In both feeder-dependent and feeder-independent conditions, cell survival was greater when Y-27632 was applied to the hESCs after sort. Specifically, treatment of cells with Y-27632 improved post-sort recovery up to four fold. To determine the long-term effects of sorting with and without the application of Y-27632, hESCs were further analyzed. Specifically, hESCs sorted with and without the addition of Y-27632 retained normal morphology, expressed hESC-specific markers as measured by immunocytochemistry and flow cytometry, and maintained a stable karyotype. In addition, the hESCs could differentiate into three germ layers in vitro and in vivo in both feeder-dependent and feeder-independent growth conditions. CONCLUSIONS/SIGNIFICANCE: The application of Y-27632 to hESCs after cell sorting improves cell recovery with no observed effect on pluripotency, and enables the consistent recovery of hESCs by FACS using multiple surface markers. This improved methodology for cell sorting of hESCs will aid many applications such as removal of hESCs from secondary cell types

  10. Time-dependent multiconfiguration self-consistent-field method based on occupation restricted multiple active space model for multielectron dynamics in intense laser fields

    CERN Document Server

    Sato, Takeshi

    2014-01-01

    The time-dependent multiconfiguration self-consistent-field method based on the occupation-restricted multiple active space model is proposed (TD-ORMAS) for multielectron dynamics in intense laser fields. Extending the previously proposed time-dependent complete-active-space self-consistent-field method [TD-CASSCF; Phys. Rev. A, {\\bf 88}, 023402 (2013)], which divides the occupied orbitals into core and active orbitals, the TD-ORMAS method {\\it further} subdivides the active orbitals into an arbitrary number of subgroups, and poses the {\\it occupation restriction} by giving the minimum and maximum number of electrons distributed in each subgroup. This enables highly flexible construction of the configuration interaction (CI) space, allowing a large-active-space simulation of dynamics, e.g., the core excitation or ionization. The equations of motion both for CI coefficients and spatial orbitals are derived based on the time-dependent variational principle, and an efficient algorithm is proposed to solve for th...

  11. Relative effectiveness of insulin pump treatment over multiple daily injections and structured education during flexible intensive insulin treatment for type 1 diabetes: cluster randomised trial (REPOSE).

    Science.gov (United States)

    2017-03-30

    Objective To compare the effectiveness of insulin pumps with multiple daily injections for adults with type 1 diabetes, with both groups receiving equivalent training in flexible insulin treatment.Design Pragmatic, multicentre, open label, parallel group, cluster randomised controlled trial (Relative Effectiveness of Pumps Over MDI and Structured Education (REPOSE) trial).Setting Eight secondary care centres in England and Scotland.Participants Adults with type 1 diabetes who were willing to undertake intensive insulin treatment, with no preference for pumps or multiple daily injections. Participants were allocated a place on established group training courses that taught flexible intensive insulin treatment ("dose adjustment for normal eating," DAFNE). The course groups (the clusters) were then randomly allocated in pairs to either pump or multiple daily injections.Interventions Participants attended training in flexible insulin treatment (using insulin analogues) structured around the use of pump or injections, followed for two years.Main outcome measures The primary outcomes were a change in glycated haemoglobin (HbA1c) values (%) at two years in participants with baseline HbA1c value of ≥7.5% (58 mmol/mol), and the proportion of participants achieving an HbA1c value of insulin dose, and episodes of moderate and severe hypoglycaemia. Ancillary outcomes included quality of life and treatment satisfaction.Results 317 participants (46 courses) were randomised (156 pump and 161 injections). 267 attended courses and 260 were included in the intention to treat analysis, of which 235 (119 pump and 116 injection) had baseline HbA1c values of ≥7.5%. Glycaemic control and rates of severe hypoglycaemia improved in both groups. The mean change in HbA1c at two years was -0.85% with pump treatment and -0.42% with multiple daily injections. Adjusting for course, centre, age, sex, and accounting for missing values, the difference was -0.24% (-2.7 mmol/mol) in favour

  12. A Novel Approach to the Fabrication of CdSe Quantum Dots in Aqueous Solution: Procedures for Controlling Size, Fluorescence Intensity, and Stability over Time

    Directory of Open Access Journals (Sweden)

    M. J. Almendral-Parra

    2014-01-01

    Full Text Available This paper report a straightforward approach for the synthesis of CdSe quantum dots (CdSe QDs in aqueous solution. This method, performed in homogeneous phase, affords optimal sizes and high quantum yields for each application desired. It is an a la carte procedure for the synthesis of nanoparticles aimed at their later application. By controlling the experimental conditions, CdSe QDs of sizes ranging between 2 and 6 nm can be obtained. The best results were achieved in an ice-bath thermostated at 4°C, using mercaptoacetic acid as dispersant. Under these conditions, a slow growth of quantum nanocrystals was generated and this was controlled kinetically by the hydrolysis of SeSO32- to generate Se2-   in situ, one of the forming species of the nanocrystal. The organic dispersant mercaptoacetate covalently binds to the Cd2+ ion, modifying the diffusion rate of the cation, and plays a key role in the stabilization of CdSe QDs. In optimum conditions, when kept in their own solution CdSe QDs remain dispersed over 4 months. The NPs obtained under optimal conditions show high fluorescence, which is a great advantage as regards their applications. The quantum efficiency is also high, owing to the formation under certain conditions of a nanoshell of Cd(OH2, values of 60% being reached.

  13. A homogeneous, high-throughput-compatible, fluorescence intensity-based assay for UDP-N-acetylenolpyruvylglucosamine reductase (MurB) with nanomolar product detection.

    Science.gov (United States)

    Shapiro, Adam B; Livchak, Stephania; Gao, Ning; Whiteaker, James; Thresher, Jason; Jahić, Haris; Huang, Jian; Gu, Rong-Fang

    2012-03-01

    A novel assay for the NADPH-dependent bacterial enzyme UDP-N-acetylenolpyruvylglucosamine reductase (MurB) is described that has nanomolar sensitivity for product formation and is suitable for high-throughput applications. MurB catalyzes an essential cytoplasmic step in the synthesis of peptidoglycan for the bacterial cell wall, reduction of UDP-N-acetylenolpyruvylglucosamine to UDP-N-acetylmuramic acid (UNAM). Interruption of this biosynthetic pathway leads to cell death, making MurB an attractive target for antibacterial drug discovery. In the new assay, the UNAM product of the MurB reaction is ligated to L-alanine by the next enzyme in the peptidoglycan biosynthesis pathway, MurC, resulting in hydrolysis of adenosine triphosphate (ATP) to adenosine diphosphate (ADP). The ADP is detected with nanomolar sensitivity by converting it to oligomeric RNA with polynucleotide phosphorylase and detecting the oligomeric RNA with a fluorescent dye. The product sensitivity of the new assay is 1000-fold greater than that of the standard assay that follows the absorbance decrease resulting from the conversion of NADPH to NADP(+). This sensitivity allows inhibitor screening to be performed at the low substrate concentrations needed to make the assay sensitive to competitive inhibition of MurB.

  14. Intensive herbicide use has selected for constitutively elevated levels of stress-responsive mRNAs and proteins in multiple herbicide-resistant Avena fatua L.

    Science.gov (United States)

    Keith, Barbara K; Burns, Erin E; Bothner, Brian; Carey, Charles C; Mazurie, Aurélien J; Hilmer, Jonathan K; Biyiklioglu, Sezgi; Budak, Hikmet; Dyer, William E

    2017-05-09

    Intensive use of herbicides has led to the evolution of two multiple herbicide-resistant (MHR) Avena fatua (wild oat) populations in Montana that are resistant to members of all selective herbicide families available for A. fatua control in US small grain crops. We used transcriptome and proteome surveys to compare constitutive changes in MHR and herbicide-susceptible (HS) plants associated with non-target site resistance. Compared to HS plants, MHR plants contained constitutively elevated levels of differentially expressed genes (DEGs) with functions in xenobiotic catabolism, stress response, redox maintenance and transcriptional regulation that are similar to abiotic stress-tolerant phenotypes. Proteome comparisons identified similarly elevated proteins including biosynthetic and multifunctional enzymes in MHR plants. Of 25 DEGs validated by RT-qPCR assay, differential regulation of 21 co-segregated with flucarbazone-sodium herbicide resistance in F3 families, and a subset of 10 of these were induced or repressed in herbicide-treated HS plants. Although the individual and collective contributions of these DEGs and proteins to MHR remain to be determined, our results support the idea that intensive herbicide use has selected for MHR populations with altered, constitutively regulated patterns of gene expression that are similar to those in abiotic stress-tolerant plants. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  15. Fluorescence spectroscopy

    DEFF Research Database (Denmark)

    Bagatolli, Luis

    2016-01-01

    Fluorescence spectroscopy is a powerful experimental tool used by scientists from many disciplines. During the last decades there have been important developments on distinct fluorescence methods, particularly those related to the study of biological phenomena. This chapter discusses...

  16. 乙醇和丙酮对绿色荧光蛋白强度作用的对比%The Comparison of the Effects of Alcohol and Acetone on Green Fluorescent Protein Intensity

    Institute of Scientific and Technical Information of China (English)

    常新; 郭泾; Yasuaki Shibata

    2005-01-01

    Objective To find out a proper way to detect green fluorescent protein (GFP). Methods Kidneys, livers and femurs from GFP transgenic mice and C57BL/6J wild type mice were employed for in vivo study.The samples were dehydrated with alcohol and acetone individually before embedding, then frozen, paraffin and resin sections were made for the detection of GFP. C3 P12 cells which derived from calvaria bone cells of GFP transgenic mouse were used for the detection of GFP in vitro. Cells were exposed to alcohol, acetone and PBS after paraformaldehyde fixation. Laser scanning microscopy was employed for GFP detection. Results In frozen sections, both kidney and liver samples which exposed to 4% buffered paraformaldehyde fixation had strong GFP signals, while GFP signal disappeared completely in fresh frozen sections without fixation. Much stronger GFP intensity was found in acetone treated samples than in alcohol treated paraffin sections, but without apparent difference in GFP intensity in acetone and alcohol treated resin samples. Acetone and alcohol made no difference in fixed C3 cells in different time courses. Conclusion Acetone treated paraffin sections are preferable for GFP detection.

  17. Common fluorescent proteins for single-molecule localization microscopy

    Science.gov (United States)

    Klementieva, Natalia V.; Bozhanova, Nina G.; Mishina, Natalie M.; Zagaynova, Elena V.; Lukyanov, Konstantin A.; Mishin, Alexander S.

    2015-07-01

    Super-resolution techniques for breaking the diffraction barrier are spread out over multiple studies nowadays. Single-molecule localization microscopy such as PALM, STORM, GSDIM, etc allow to get super-resolved images of cell ultrastructure by precise localization of individual fluorescent molecules via their temporal isolation. However, these methods are supposed the use of fluorescent dyes and proteins with special characteristics (photoactivation/photoconversion). At the same time, there is a need for retaining high photostability of fluorophores during long-term acquisition. Here, we first showed the potential of common red fluorescent protein for single-molecule localization microscopy based on spontaneous intrinsic blinking. Also, we assessed the effect of different imaging media on photobleaching of these fluorescent proteins. Monomeric orange and red fluorescent proteins were examined for stochastic switching from a dark state to a bright fluorescent state. We studied fusions with cytoskeletal proteins in NIH/3T3 and HeLa cells. Imaging was performed on the Nikon N-STORM system equipped with EMCCD camera. To define the optimal imaging conditions we tested several types of cell culture media and buffers. As a result, high-resolution images of cytoskeleton structure were obtained. Essentially, low-intensity light was sufficient to initiate the switching of tested red fluorescent protein reducing phototoxicity and provide long-term live-cell imaging.

  18. Highly fluorescent gold nanoclusters based sensor for the detection of quercetin

    Energy Technology Data Exchange (ETDEWEB)

    Chen Zhanguang, E-mail: kqlu@stu.edu.cn; Qian Sihua [Shantou University, Department of Chemistry (China); Chen Junhui, E-mail: chenjupush@126.com [Peking University Shenzhen Hospital, Interventional Oncology and Minimally Invasive Therapies Department (China); Chen Xi [Guangdong Pharmaceutical University (China)

    2012-12-15

    In this contribution, novel luminescent gold nanoclusters were synthesized by utilizing bovine serum albumin as templates with a simple, rapid, and one-pot procedure. The as-prepared gold nanoclusters were highly dispersed in aqueous solution and emitted an intense red fluorescence under UV light (365 nm). They exhibited strong fluorescence and the maximum excitation and emission wavelengths were 480 and 613.5 nm. In addition, the bovine serum albumin-stabilized gold nanoclusters were successfully utilized as novel fluorescent probes for the detection of quercetin for the first time. It was found that the addition of quercetin induced the strong fluorescence intensity of the gold nanoclusters to decrease. The decrease in fluorescence intensity of the gold nanoclusters caused by quercetin allowed the sensitive detection of quercetin in the range of 8.9 Multiplication-Sign 10{sup -8}-1.8 Multiplication-Sign 10{sup -4} mol L{sup -1}. The detection limit for quercetin is 1.8 Multiplication-Sign 10{sup -8} mol L{sup -1} at a signal-to-noise ratio of 3. The present sensor for quercetin detection possessed a low detection limit and wide linear range. In addition, the real samples were analyzed with satisfactory results.

  19. Use of O-J-I-P Chlorophyll Fluorescence Transients to Probe Multiple Effects of UV-C Radiation on the Photosynthetic Apparatus of Euglena

    Directory of Open Access Journals (Sweden)

    Chalinda Koshitha Beneragama

    2014-12-01

    Full Text Available Although the kinetic chlorophyll fluorescence signals are rich in information, most of the chlorophyll fluorescence related studies deal only with the quantum yield of primary photochemistry (Fv/Fm. JIP-test based OJIP fluorescence transient analysis is relatively a new technique to investigate the environmental stress responses of photosynthetic organisms. In the present study, the deleterious effects of ultraviolet (UV radiation on the photosynthetic machinery were probed by the JIP-test in Euglena, one of the most potent organisms for the future space stations. The cells were exposed to a series of UV-C doses and immediately after exposure, survival percentage was determined with Neutral Red staining, and the chlorophyll fluorescence was measured using AquaPen AP-C 100 fluorometer. Resultant OJIP transients were analyzed according to JIP-test, and several functional and structural parameters were derived to explain the PSII behavior. Results indicated that the UV-C induced inhibition of electron transport is severely affected due to higher sensitivity of dark reactions after QA -, represented as ψo, the electron transfer probability, than of the light dependent reactions, represented as φPo, the trapping probability. The performance index (PIABS of PSII, which is a combination of the indices of three independent parameters, decreased markedly in exponential manner in response to UVC. Results illustrate the advantage of using a number of fluorescent parameters over the use of one parameter, often the Fv/Fm.

  20. Influence of two-electrode montages on the level-specific (LS) CE-Chirp auditory brainstem response (ABR) at multiple intensity levels.

    Science.gov (United States)

    Dzulkarnain, Ahmad Aidil Arafat; Noor Ibrahim, Siti Hajra Mu'minah; Anuar, Nur Farah Aida; Abdullah, Siti Aisyah; Tengku Zam Zam, Tengku Zulaila Hasma; Rahmat, Sarah; Mohd Ruzai, Muhammad Amar

    2017-10-01

    To investigate the influence of two different electrode montages (ipsilateral: reference to mastoid and vertical: reference to nape of neck) to the ABR results recorded using a level-specific (LS)-CE-Chirp® in normally hearing subjects at multiple intensities levels. Quasi-experimental and repeated measure study designs were applied in this study. Two different stopping criteria were used, (1) a fixed-signal averaging 4000 sweeps and, (2) a minimum quality indicator of Fmp = 3.1 with a minimum of 800 sweeps. Twenty-nine normally hearing adults (18 females, 11 male) participated. Wave V amplitudes were significantly larger in the LS CE-Chirp® recorded from the vertical montage than the ipsilateral montage. Waves I and III amplitudes were significantly larger from the ipsilateral LS CE-Chirp® than from the other montages and stimulus combinations. The differences in the quality of the ABR recording between the vertical and ipsilateral montages were marginal. Overall, the result suggested that the vertical LS CE-Chirp® ABR had a high potential for a threshold-seeking application, because it produced a higher wave V amplitude. The Ipsilateral LS CE-Chirp® ABR, on the other hand, might also have a high potential for the site of lesion application, because it produced larger waves I and III amplitudes.

  1. A phase 2 study of three low-dose intensity subcutaneous bortezomib regimens in elderly frail patients with untreated multiple myeloma.

    Science.gov (United States)

    Larocca, A; Bringhen, S; Petrucci, M T; Oliva, S; Falcone, A P; Caravita, T; Villani, O; Benevolo, G; Liberati, A M; Morabito, F; Montefusco, V; Passera, R; De Rosa, L; Omedé, P; Vincelli, I D; Spada, S; Carella, A M; Ponticelli, E; Derudas, D; Genuardi, M; Guglielmelli, T; Nozzoli, C; Aghemo, E; De Paoli, L; Conticello, C; Musolino, C; Offidani, M; Boccadoro, M; Sonneveld, P; Palumbo, A

    2016-06-01

    This phase 2 trial evaluated three low-dose intensity subcutaneous bortezomib-based treatments in patients ⩾75 years with newly diagnosed multiple myeloma (MM). Patients received subcutaneous bortezomib plus oral prednisone (VP, N=51) or VP plus cyclophosphamide (VCP, N=51) or VP plus melphalan (VMP, N=50), followed by bortezomib maintenance, and half of the patients were frail. Response rate was 64% with VP, 67% with VCP and 86% with VMP, and very good partial response rate or better was 26%, 28.5% and 49%, respectively. Median progression-free survival was 14.0, 15.2 and 17.1 months, and 2-year OS was 60%, 70% and 76% in VP, VCP, VMP, respectively. At least one drug-related grade ⩾3 non-hematologic adverse event (AE) occurred in 22% of VP, 37% of VCP and 33% of VMP patients; the discontinuation rate for AEs was 12%, 14% and 20%, and the 6-month rate of toxicity-related deaths was 4%, 4% and 8%, respectively. The most common grade ⩾3 AEs included infections (8-20%), and constitutional (10-14%) and cardiovascular events (4-12%); peripheral neuropathy was limited (4-6%). Bortezomib maintenance was effective and feasible. VP, VCP and VMP regimens demonstrated no substantial difference. Yet, toxicity was higher with VMP, suggesting that a two-drug combination followed by maintenance should be preferred in frail patients.

  2. Signal intensity change on unenhanced T1-weighted images in dentate nucleus following gadobenate dimeglumine in patients with and without previous multiple administrations of gadodiamide

    Energy Technology Data Exchange (ETDEWEB)

    Ramalho, Joana [University of North Carolina Hospital, Chapel Hill, NC (United States); Centro Hospitalar de Lisboa Central, Lisbon (Portugal); Semelka, Richard C.; Castillo, Mauricio [University of North Carolina Hospital, Chapel Hill, NC (United States); AlObaidy, Mamdoh [University of North Carolina Hospital, Chapel Hill, NC (United States); King Faisal Specialist Hospital and Research Center, Riyadh (Saudi Arabia); Ramalho, Miguel [University of North Carolina Hospital, Chapel Hill, NC (United States); Hospital Garcia de Orta, Almada (Portugal); Nunes, Renato H. [University of North Carolina Hospital, Chapel Hill, NC (United States); Santa Casa de Misericordia de Sao Paulo, Sao Paulo (Brazil)

    2016-11-15

    To evaluate the impact of previous administration of gadodiamide and neural tissue gadolinium deposition in patients who received gadobenate dimeglumine. Our population included 62 patients who underwent at least three administrations of gadobenate dimeglumine, plus an additional contrast-enhanced last MRI for reference, divided into two groups: group 1, patients who in addition to gadobenate dimeglumine administrations had prior exposure to multiple doses of gadodiamide; group 2, patients without previous exposure to other gadolinium-based contrast agent (GBCAs). Quantitative analysis was performed on the first and last gadobenate dimeglumine MRIs in both groups. Dentate nucleus-to-middle cerebellar peduncle signal intensity ratios (DN/MCP) and relative change (RC) in signal over time were calculated and compared between groups using generalized additive model. Group 1 showed significant increase in baseline and follow-up DN/MCP compared to group 2 (p < 0.0001). The RC DN/MCP showed a non-statistically significant trend towards an increase in patients who underwent previous gadodiamide (p = 0.0735). There is increased T1 signal change over time in patients who underwent gadobenate dimeglumine and had received prior gadodiamide compared to those without known exposure to previous gadodiamide. A potentiating effect from prior gadodiamide on subsequent administered gadobenate dimeglumine may occur. (orig.)

  3. Factors affecting the implementation of complex and evolving technologies: multiple case study of intensity-modulated radiation therapy (IMRT in Ontario, Canada

    Directory of Open Access Journals (Sweden)

    Bak Kate

    2011-07-01

    Full Text Available Abstract Background Research regarding the decision to adopt and implement technological innovations in radiation oncology is lacking. This is particularly problematic since these technologies are often complex and rapidly evolving, requiring ongoing revisiting of decisions regarding which technologies are the most appropriate to support. Variations in adoption and implementation decisions for new radiation technologies across cancer centres can impact patients' access to appropriate and innovative forms of radiation therapy. This study examines the key steps in the process of adopting and implementing intensity modulated radiation therapy (IMRT in publicly funded cancer centres and identifies facilitating or impeding factors. Methods A multiple case study design, utilizing document analysis and key informant interviews was employed. Four cancer centres in Ontario, Canada were selected and interviews were conducted with radiation oncologists, medical physicists, radiation therapists, and senior administrative leaders. Results Eighteen key informants were interviewed. Overall, three centres made fair to excellent progress in the implementation of IMRT, while one centre achieved only limited implementation as of 2009. Key factors that influenced the extent of IMRT implementation were categorized as: 1 leadership, 2 training, expertise and standardization, 3 collaboration, 4 resources, and 5 resistance to change. Conclusion A framework for the adoption and implementation of complex and evolving technologies is presented. It identifies the key factors that should be addressed by decision-makers at specific stages of the adoption/implementation process.

  4. Mean frequency and relative fluorescence intensity measurement of γ-H2AX foci dose response in PBL exposed to γ-irradiation: An inter- and intra-laboratory comparison and its relevance for radiation triage.

    Science.gov (United States)

    Venkateswarlu, Raavi; Tamizh, Selvan G; Bhavani, Manivannan; Kumar, Arun; Alok, Amit; Karthik, Kanagaraj; Kalra, Namita; Vijayalakshmi, J; Paul, Solomon F D; Chaudhury, N K; Venkatachalam, Perumal

    2015-12-01

    Measurement of γ-H2AX protein changes in the peripheral blood lymphocytes (PBL) of individuals exposed to ionizing radiation is a simple, sensitive, and rapid assay for radiation triage and early marker of dose estimation. The qualitative and quantitative measurements of the protein changes were examined using flow cytometry and microscopy. Whole blood and isolated lymphocytes were exposed in vitro between 0.1 and 5 Gy doses of (60) Co γ-radiation at a dose rate of 1 Gy/min. Radiation induced γ-H2AX foci frequency (n = 3) and relative fluorescence intensity (n = 7) in PBL was measured at 0.5 and 2 hrs postexposure. The observed dose response for γ-H2AX foci frequency at both time points, for whole blood and isolated lymphocytes did not show any significant (P > 0.05) differences. However, when compared with γ-H2AX foci frequency scored manually (microscopy), the semiautomated analysis (captured images) showed a better correlation (r(2) = 0.918) than that obtained with automated (Metafer) scoring (r(2) = 0.690). It is noteworthy to mention that, the γ-H2AX foci frequency quantified using microscopy showed a dose dependent increase up to 2 Gy and the relative fluorescence intensity (RFI) measured with flow cytometry revealed an increase up to 5 Gy in the PBL exposed in vitro. Moreover, a better correlation was observed between the γ-H2AX foci frequency obtained by manual scoring and RFI (r(2) = 0.910). Kinetic studies showed that the γ-H2AX foci remain more or less unchanged up to 4 hrs and reduces gradually over 48 hrs of postexposure at 37°C. Further, inter and intra-laboratory comparisons showed consistency in the scoring of γ-H2AX foci frequency by manual and semiautomated scoring. The overall results suggest that measurement of γ-H2AX (microscopy and flow cytometry) should be employed within 4 to 6 hrs for a reliable dosimetry either by sharing the work load between the laboratories or investing more manpower; however, triage can be possible even up

  5. Design and daytime performance of laser-induced fluorescence spectrum lidar for simultaneous detection of multiple components, dissolved organic matter, phycocyanin, and chlorophyll in river water.

    Science.gov (United States)

    Saito, Yasunori; Kakuda, Kei; Yokoyama, Mizuho; Kubota, Tomoki; Tomida, Takayuki; Park, Ho-Dong

    2016-08-20

    In this work, we developed mobile laser-induced fluorescence spectrum (LIFS) lidar based on preliminary experiments on the excitation emission matrix of a water sample and a method for reducing solar background light using the synchronous detection technique. The combination of a UV short-pulse laser (355 nm, 6 ns) for fluorescence excitation with a 10-100 ns short-time synchronous detection using a gated image-intensified multi-channel CCD of the fluorescence made the LIFS lidar operation possible even in daytime. The LIFS lidar with this construction demonstrated the potential of natural river/lake water quality monitoring at the Tenryu River/Lake Suwa. Three main components in the fluorescence data of the water, dissolved organic matter, phycocyanin, and chlorophyll, were extracted by spectral analysis using the standard spectral functions of these components. Their concentrations were estimated by adapting experimentally calibrated data. Results of long-term field observations using our LIFS lidar from 2010 to 2012 show the necessity of simultaneous multi-component detection to understand the natural water environment.

  6. Diffusion behavior of the fluorescent proteins eGFP and Dreiklang in solvents of different viscosity monitored by fluorescence correlation spectroscopy

    Science.gov (United States)

    Junghans, Cornelia; Schmitt, Franz-Josef; Vukojević, Vladana; Friedrich, Thomas

    2016-12-01

    Fluorescence correlation spectroscopy relies on temporal autocorrelation analysis of fluorescence intensity fluctuations that spontaneously arise in systems at equilibrium due to molecular motion and changes of state that cause changes in fluorescence, such as triplet state transition, photoisomerization and other photophysical transformations, to determine the rates of these processes. The stability of a fluorescent molecule against dark state conversion is of particular concern for chromophores intended to be used as reference tags for comparing diffusion processes on multiple time scales. In this work, we analyzed properties of two fluorescent proteins, the photoswitchable Dreiklang and its parental eGFP, in solvents of different viscosity to vary the diffusion time through the observation volume element by several orders of magnitude. In contrast to eGFP, Dreiklang undergoes a dark-state conversion on the time scale of tens to hundreds of microseconds under conditions of intense fluorescence excitation, which results in artificially shortened diffusion times if the diffusional motion through the observation volume is sufficiently slowed down. Such photophysical quenching processes have also been observed in FCS studies on other photoswitchable fluorescent proteins including Citrine, from which Dreiklang was derived by genetic engineering. This property readily explains the discrepancies observed previously between the diffusion times of eGFP- and Dreiklang-labeled plasma membrane protein complexes.

  7. Phasor approaches simplify the analysis of tryptophan fluorescence data in protein denaturation studies

    NARCIS (Netherlands)

    Bader, A.N.; Visser, N.V.; Amerongen, van H.; Visser, A.J.W.G.

    2014-01-01

    The intrinsic fluorescence of tryptophan is frequently used to investigate the structure of proteins. The analysis of tryptophan fluorescence data is challenging: fluorescence (anisotropy) decays typically have multiple lifetime (correlation time) components and fluorescence spectra are broad and ex

  8. Fluorescence Studies of Selected 2-Alkylaminopyrimidines

    Directory of Open Access Journals (Sweden)

    B. K. Low

    2004-06-01

    Full Text Available The reactions of 2-chloropyrimidine with methylamine, ethylamine and piperidine gave the corresponding 2-N-methylamino-, 2-N-ethylamino- and 2N- piperidinopyrimidines, respectively. The fluorescence properties of these alkylamino derivatives in chloroform, ethyl acetate, carbon tetrachloride, acetone, ether, ethanol and methanol were studied. All the alkylamino derivatives showed the highest fluorescence intensity in polar protic solvents; thus 2-N-methylaminopyrimidine (highest fluorescence intensity at 377 nm when excited at 282 nm and 2-N-ethylaminopyrimidine (highest fluorescence intensity at 375 nm, when excited at 286 nm showed the highest fluorescence in methanol. In ethanol, 2-N-piperidinopyrimidine showed a fluorescence peak at 403 nm when excited at 360 nm and in chloroform it fluoresced at 392 nm when excited at 356 nm.

  9. Technique of Hadamard transform microscope fluorescence image analysis

    Institute of Scientific and Technical Information of China (English)

    梅二文; 顾文芳; 曾晓斌; 陈观铨; 曾云鹗

    1995-01-01

    Hadamard transform spatial multiplexed imaging technique is combined with fluorescence microscope and an instrument of Hadamard transform microscope fluorescence image analysis is developed. Images acquired by this instrument can provide a lot of useful information simultaneously, including three-dimensional Hadamard transform microscope cell fluorescence image, the fluorescence intensity and fluorescence distribution of a cell, the background signal intensity and the signal/noise ratio, etc.

  10. Characterization of obstetric patients with multiple organ failure in the intensive care unit of a Havana teaching hospital, 1998 to 2006.

    Science.gov (United States)

    Pérez, Albadio; Acevedo, Omar; Tamayo, Felicia del Consuelo; Oviedo, Regino

    2010-01-01

    Most obstetric patients admitted to intensive care units (ICU) present life-threatening complications of pregnancy, delivery, and postpartum, often leading to multiple organ failure (MOF), considered the main cause of death in ICUs. Although maternal mortality is an important indicator of health status in women and nations, few studies have analyzed MOF in obstetric ICU patients. The Sequential Organ Failure Assessment (SOFA) scale is a prognostic tool for ranking organ dysfunction in critically ill patients and correlating scores with outcome. Characterize obstetric patients diagnosed with MOF in the ICU of the Enrique Cabrera General Teaching Hospital, Havana, Cuba, between January 1, 1998 and December 31, 2006. A descriptive observational study was conducted of obstetric patients admitted to the ICU for >24 hours during the study period and diagnosed with MOF. Of 422 obstetric patients admitted, 58 met inclusion criteria. Patients' clinical characteristics, chronic diseases, diagnoses, Acute Physiology and Chronic Health Evaluation (APACHE II) scores, SOFA scores and discharge status were recorded in a data collection form. Day 1, Day 3, and maximum APACHE II and SOFA scores were calculated, and 3 prognostic groups were formed based on the difference between SOFA Day 1 and Day 3 scores (SOFAd 3-1: 0, poor prognosis). Data was entered in an EXCEL database and processed using SPSS 13.0 software. Quantitative variables were described by mean and standard deviation; qualitative variables by totals and percentages. A chi-square test was used to analyze associations between these variables and discharge status (alive or deceased) with significance level p 0 score with an unfavorable prognosis (84.0% mortality). Respiratory and cardiovascular systems were the most affected (75.9% each). Further studies analyzing MOF in obstetric patients are urgently needed in order to adopt more effective strategies for reducing maternal mortality in Cuba.

  11. Quantitative evaluation of local pulmonary distribution of TiO2 in rats following single or multiple intratracheal administrations of TiO2 nanoparticles using X-ray fluorescence microscopy.

    Science.gov (United States)

    Zhang, Guihua; Shinohara, Naohide; Kano, Hirokazu; Senoh, Hideki; Suzuki, Masaaki; Sasaki, Takeshi; Fukushima, Shoji; Gamo, Masashi

    2016-10-01

    Uneven pulmonary nanoparticle (NP) distribution has been described when using single-dose intratracheal administration tests. Multiple-dose intratracheal administrations with small quantities of NPs are expected to improve the unevenness of each dose. The differences in local pulmonary NP distribution (called microdistribution) between single- and multiple-dose administrations may cause differential pulmonary responses; however, this has not been evaluated. Here, we quantitatively evaluated the pulmonary microdistribution (per mesh: 100 μm × 100 μm) of TiO2 in lung sections from rats following one, two, three, or four doses of TiO2 NPs at a same total dosage of 10 mg kg(-1) using X-ray fluorescence microscopy. The results indicate that: (i) multiple-dose administrations show lower variations in TiO2 content (ng mesh(-1) ) for sections of each lobe; (ii) TiO2 appears to be deposited more in the right caudal and accessory lobes located downstream of the administration direction of NP suspensions, and less so in the right middle lobes, irrespective of the number of doses; (iii) there are not prominent differences in the pattern of pulmonary TiO2 microdistribution between rats following single and multiple doses of TiO2 NPs. Additionally, the estimation of pulmonary TiO2 deposition for multiple-dose administrations imply that every dose of TiO2 would be randomly deposited only in part of the fixed 30-50% of lung areas. The evidence suggests that multiple-dose administrations do not offer remarkable advantages over single-dose administration on the pulmonary NP microdistribution, although multiple-dose administrations may reduce variations in the TiO2 content for each lung lobe. Copyright © 2016 John Wiley & Sons, Ltd.

  12. A fluorescence switch based on a controllable photochromic naphthopyran group

    Energy Technology Data Exchange (ETDEWEB)

    Chen Lizhen [Faculty of Chemistry, Northeast Normal University, Changchun 130024 (China); Wang Guang, E-mail: wangg923@nenu.edu.cn [Faculty of Chemistry, Northeast Normal University, Changchun 130024 (China); Zhao Xiancai [Faculty of Chemistry, Northeast Normal University, Changchun 130024 (China)

    2011-08-15

    A fluorescence switch based on photoisomerization of naphthopyran (NP) has been designed by employing 2-(pyridin-2-yl)-benzimidazole (BPI) and the naphthopyran containing two pyran rings (NP) as fluorescent dye and photochromic compound, respectively. The fluorescence switch of benzimidazole derivative can be modulated either by controlling the irradiation time of UV light or by adjusting the amount ratio of fluorescent benzimidazole derivative to photochromic naphthopyran in both solution and polymethyl methacrylate (PMMA) film. The experimental results indicated that the decrease of fluorescence intensity of benzimidazole derivative is attributed to the interaction of benzimidazole with naphthopyran. - Highlights: > Naphthopyran was first used to fabricate fluorescence switch with benzimidazole derivative. > Fluorescence intensity can be modulated by controlling the UV irradiation time. > Fluorescence intensity can be adjusted by changing the ratio of benzimidazole derivative to naphthopyran. > Decrease of fluorescence intensity is attributed to the interaction of benzimidazole derivative and naphthopyran.

  13. Flow cytometric fluorescence lifetime analysis of DNA binding fluorochromes

    Energy Technology Data Exchange (ETDEWEB)

    Crissman, Harry A.; Cui, H. H. (H. Helen); Steinkamp, J. A.

    2002-01-01

    Most flow cytometry (FCM) applications monitor fluorescence intensity to quantitate the various cellular parameters; however, the fluorescence emission also contains information relative to the fluorescence lifetime. Recent developments in FCM (Pinsky et al., 1993; Steinkamp & Crissman, 1993; Steinkamp et al., 1993), provide for the measurement of fluorescence lifetime which is also commonly referred to as fluorescence decay, or the time interval in which a fluorochrome remains in the excited state. Many unbound fluorochromes have characteristic lifetime values that are determined by their molecular structure; however, when the probe becomes bound, the lifetime value is influenced by a number of factors that affect the probe interaction with a target molecule. Monitoring the changes in the lifetime of the probe yields information relating to the molecular conformation, the functional state or activity of the molecular target. In addition, the lifetime values can be used as signatures to resolve the emissions of multiple fluorochrome labels with overlapping emission spectra that cannot be resolved by conventional FCM methodology. Such strategies can increase the number of fluorochrome combinations used in a flow cytometer with a single excitation source. Our studies demonstrate various applications of lifetime measurements for the analysis of the binding of different fluorochromes to DNA in single cells. Data presented in this session will show the utility of lifetime measurements for monitoring changes in chromatin structure associated with cell cycle progression, cellular differentiation, or DNA damage, such as induced during apoptosis. Several studies show that dyes with specificity for nucleic acids display different lifetime values when bound to DNA or to dsRNA. The Phase Sensitive Flow Cytometer is a multiparameter instrument, capable of performing lifetime measurements in conjunction with all the conventional FCM measurements. Future modifications of this

  14. Nine New Fluorescent Probes

    Science.gov (United States)

    Lin, Tsung-I.; Jovanovic, Misa V.; Dowben, Robert M.

    1989-06-01

    Absorption and fluorescence spectroscopic studies are reported here for nine new fluorescent probes recently synthesized in our laboratories: four pyrene derivatives with substituents of (i) 1,3-diacetoxy-6,8-dichlorosulfonyl, (ii) 1,3-dihydroxy-6,8-disodiumsulfonate, (iii) 1,3-disodiumsulfonate, and (iv) l-ethoxy-3,6,8-trisodiumsulfonate groups, and five [7-julolidino] coumarin derivatives with substituents of (v) 3-carboxylate-4-methyl, (vi) 3- methylcarboxylate, (vii) 3-acetate-4-methyl, (viii) 3-propionate-4-methyl, and (ix) 3-sulfonate-4-methyl groups. Pyrene compounds i and ii and coumarin compounds v and vi exhibit interesting absorbance and fluorescence properties: their absorption maxima are red shifted compared to the parent compound to the blue-green region, and the band width broadens considerably. All four blue-absorbing dyes fluoresce intensely in the green region, and the two pyrene compounds emit at such long wavelengths without formation of excimers. The fluorescence properties of these compounds are quite environment-sensitive: considerable spectral shifts and fluorescence intensity changes have been observed in the pH range from 3 to 10 and in a wide variety of polar and hydrophobic solvents with vastly different dielectric constants. The high extinction and fluorescence quantum yield of these probes make them ideal fluorescent labeling reagents for proteins, antibodies, nucleic acids, and cellular organelles. The pH and hydrophobicity-dependent fluorescence changes can be utilized as optical pH and/or hydrophobicity indicators for mapping environmental difference in various cellular components in a single cell. Since all nine probes absorb in the UV, but emit at different wavelengths in the visible, these two groups of compounds offer an advantage of utilizing a single monochromatic light source (e.g., a nitrogen laser) to achieve multi-wavelength detection for flow cytometry application. As a first step to explore potential application in

  15. IMPACT OF PRETRANSPLANT DONOR AND RECIPIENT CYTOMEGALOVIRUS SEROSTATUS ON OUTCOME FOR MULTIPLE MYELOMA PATIENTS UNDERGOING REDUCED INTENSITY CONDITIONING ALLOGENEIC STEM CELL TRANSPLANTATION.

    Directory of Open Access Journals (Sweden)

    Jean Elcheikh

    2013-04-01

    Full Text Available To investigate the impact of pre-transplant CMV serostatus of donor or recipient on outcome of patients undergoing allogeneic hematopoietic stem cell transplantation (Allo-SCT for Multiple Myeloma (MM. To our knowledge no data are available in the literature about this issue. We retrospectively followed 99 consecutive patients who underwent reduced-intensity conditioning (RIC Allo-SCT for MM in our cancer centre at Marseille between January 2000 and January 2012. Based upon CMV serostatus, patients were classified as low risk (donor [D]-/recipient [R]- 17 patients (17.1%, intermediate risk (D+/R 14 patients (14.1%, or high risk – either (D-/R+ 31 patients (31.3% or (D+/R+, 37 patients (37.3%. Cumulative incidence of CMV reactivation was 39% with a median time of 61 days (26–318. Three patients (3% developed CMV disease. Two factors were associated with CMV reactivation: CMV serostatus group (low: 0% vs intermediate: 29% vs high: 50%; p=0.001 and the presence of grade II–IV acute GvHD (Hazard Ratio: HR=2.1 [1.1–3.9]. Thirty-six of the 39 patients (92% with CMV reactivation did not present positive detection of CMV after a 21-day median duration preemptive treatment with ganciclovir. Cumulative incidence of day 100 grade II–IV acute GvHD, 1-year chronic GvHD and day 100 transplantation related mortality (TRM were 37%, 36% and 9%, respectively. CMV reactivation and serostatus were not associated with increased GvHD and TRM or short survival. Only the presence of acute GvHD as a time dependent variable was significantly associated with increased TRM (p=0.005. Two-year overall and progression free survival were 56% and 34%, respectively. Donor and recipient CMV serostatus and acute GvHD are independent factors for increased CMV reactivation in high-risk MM patients undergoing RIC Allo-SCT. However, we did not find any influence of CMV reactivation on post transplantation outcome. CMV monitoring and pre-emptive treatment strategy could in

  16. ICU在严重多发伤救治中的作用%Role of intensive care unit in treatment of severe multiple trauma

    Institute of Scientific and Technical Information of China (English)

    施建国; 姚远; 周继红; 李勇; 骆建军; 侯振海; 邱俊; 张良

    2013-01-01

    Objective To investigate the criteria and significance for management of severe multiple trauma in the intensive care unit (ICU).Methods Data of 64 patients with non-fatal severe multi-trauma (ISS≥20) treated in the First Aid Center for Traffic Injuries in 117th Hospital of PLA between 2006-2012 documented in China Trauma Database were analyzed.Differences in trauma scoring,complications,functional prognosis and medical cost were analyzed between ICU and non-ICU patients.Results There were statistical differences for GCS and revised trauma score (RTS) between ICU patients and non-ICU patients (P < 0.05),but the differences were insignificant for ISS and new ISS (NISS).Likewise,the differences were significant for complication incidence and functional independence measure (FIM) on discharge (P < 0.05),but not in the length of hospital stay and medical cost.Complication incidence of non-ICU patients was 2.96 times that of ICU patients.FIM of ICU patients scored 10.11 ± 2.10,far higher than that of non-ICU patients (P < 0.05).Conclusion For severe multiple trauma patients with ISS≥20,the admission to the ICU is helpful for reduction of the complication rate.%目的 探讨ICU治疗严重多发伤的伤情标准及意义. 方法 通过分析“中华创伤数据库”收录的解放军第一一七医院交通伤急救中心2006-2012年收治的64例(ISS≥20分)非死亡严重多发伤患者资料,比较ICU救治组和非ICU救治组在创伤评分、并发症、功能预后及医疗成本等方面的差异. 结果 两组患者ISS与新损伤严重度评分(new ISS,NISS)差异无统计学意义,但GCS和修正创伤记分(revised trauma score,RTS)差异有统计学意义(P<0.05);两组患者住院天数和医疗费用等差异无统计学意义,但发生并发症例数和出院时功能独立性评定(functional independence measure,FIM)差异有统计学意义(P<0.05).非ICU救治组发生并发症的可能性是ICU救治组的2.96

  17. Fluorescent sensors based on bacterial fusion proteins

    Science.gov (United States)

    Prats Mateu, Batirtze; Kainz, Birgit; Pum, Dietmar; Sleytr, Uwe B.; Toca-Herrera, José L.

    2014-06-01

    Fluorescence proteins are widely used as markers for biomedical and technological purposes. Therefore, the aim of this project was to create a fluorescent sensor, based in the green and cyan fluorescent protein, using bacterial S-layers proteins as scaffold for the fluorescent tag. We report the cloning, expression and purification of three S-layer fluorescent proteins: SgsE-EGFP, SgsE-ECFP and SgsE-13aa-ECFP, this last containing a 13-amino acid rigid linker. The pH dependence of the fluorescence intensity of the S-layer fusion proteins, monitored by fluorescence spectroscopy, showed that the ECFP tag was more stable than EGFP. Furthermore, the fluorescent fusion proteins were reassembled on silica particles modified with cationic and anionic polyelectrolytes. Zeta potential measurements confirmed the particle coatings and indicated their colloidal stability. Flow cytometry and fluorescence microscopy showed that the fluorescence of the fusion proteins was pH dependent and sensitive to the underlying polyelectrolyte coating. This might suggest that the fluorescent tag is not completely exposed to the bulk media as an independent moiety. Finally, it was found out that viscosity enhanced the fluorescence intensity of the three fluorescent S-layer proteins.

  18. Fluorescence spectral studies of Gum Arabic: Multi-emission of Gum Arabic in aqueous solution

    Energy Technology Data Exchange (ETDEWEB)

    Dhenadhayalan, Namasivayam, E-mail: ndhena@gmail.com [Department of Chemistry, National Taiwan University, Taipei, Taiwan (China); Mythily, Rajan, E-mail: rajanmythily@gmail.com [Department of Chemistry, Dwaraka Doss Goverdhan Doss Vaishnav College (Autonomous), 833, Gokul Bagh, E.V.R. Periyar Road, Arumbakkam, Chennai 600 106 (India); Kumaran, Rajendran, E-mail: kumaranwau@rediffmail.com [Department of Chemistry, Dwaraka Doss Goverdhan Doss Vaishnav College (Autonomous), 833, Gokul Bagh, E.V.R. Periyar Road, Arumbakkam, Chennai 600 106 (India)

    2014-11-15

    Gum Arabic (GA), a food hydrocolloid is a natural composite obtained from the stems and branches of Acacia Senegal and Acacia Seyal trees. GA structure is made up of highly branched arabinogalactan polysaccharides. Steady-state absorption, fluorescence, and time-resolved fluorescence spectral studies of acid hydrolyzed GA solutions were carried out at various pH conditions. The fluorescence in GA is predominantly attributed to the presence of tyrosine and phenylalanine amino acids. The presence of multi-emissive peaks at different pH condition is attributed to the exposure of the fluorescing amino acids to the aqueous phase, which contains several sugar units, hydrophilic and hydrophobic moieties. Time-resolved fluorescence studies of GA exhibits a multi-exponential decay with different fluorescence lifetime of varying amplitude which confirms that tyrosine is confined to a heterogeneous microenvironment. The existence of multi-emissive peaks with large variation in the fluorescence intensities were established by 3D emission contour spectral studies. The probable location of the fluorophore in a heterogeneous environment was further ascertained by constructing a time-resolved emission spectrum (TRES) and time-resolved area normalized emission spectrum (TRANES) plots. Fluorescence spectral technique is used as an analytical tool in understanding the photophysical properties of a water soluble complex food hydrocolloid containing an intrinsic fluorophore located in a multiple environment is illustrated. - Highlights: • The Manuscript deals with the steady state absorption, emission, fluorescence lifetime and time-resolved emission spectrum studies of Gum Arabic in aqueous medium at various pH conditions. • The fluorescence emanates from the tyrosine amino acid present in GA. • Change in pH results in marked variation in the fluorescence spectral properties of tyrosine. • Fluorescence spectral techniques are employed as a tool in establishing the

  19. Changes in the fluorescence composition of multiple DOM sources over pH gradients assessed by combining parallel factor analysis and self-organizing maps

    Science.gov (United States)

    Cuss, C. W.; Shi, Y. X.; McConnell, S. M.; Guéguen, C.

    2014-09-01

    Dissolved organic matter is a ubiquitous constituent of natural waters that plays key roles in several important processes. The fluorescence properties of DOM have been linked to its functionality, but these properties may vary with pH. In this study Kohonen's self-organizing maps (SOMs) were applied to excitation-emission matrices (EEMs) of fresh dissolved organic matter (DOM) from three sources: senescent sugar-maple leaves and white spruce needles, and humified white spruce needles, over a pH range of ~4.5 - 12.5. SOMs were applied to raw EEMs, EEMs reduced in dimensionality by pre-processing using parallel factor analysis (PARAFAC), and PARAFAC loading proportions normalized to values at initial pH. Some separation of EEMs into source-based clusters was achieved in the SOM of raw EEMs, but commingling was apparent and evidence of changes over pH gradients was overshadowed. SOMs of PARAFAC component proportions demonstrated clear source-based clustering, and pH-based gradients were visible for DOM from senescent and humified spruce needles. Changes in optical properties were obvious over pH gradients in the SOM of components normalized to starting condition. Component proportions decreased to values as low as 5% of the initial values for microbial humic-like peak M and increased to as high as 278% for a humic-like component. Tyrosine-like fluorescence increased to 112% of initial over increasing pH in humified spruce leachates but decreased to as low as 45% in the other leachates. The combination of PARAFAC and SOM drastically enhanced visualization and interpretability of pH-induced changes in DOM compared to either method alone.

  20. New fluorescence parameters for monitoring photosynthesis in plants

    NARCIS (Netherlands)

    Force, L.; Critchley, Ch.; Rensen, van J.J.S.

    2003-01-01

    Chlorophyll fluorescence measurements have a wide range of applications from basic understanding of photosynthesis functioning to plant environmental stress responses and direct assessments of plant health. The measured signal is the fluorescence intensity (expressed in relative units) and the most

  1. Evidence of Multiple Treponema Phylotypes Involved in Bovine Digital Dermatitis as Shown by 16S rRNA Gene Analysis and Fluorescence In Situ Hybridization

    DEFF Research Database (Denmark)

    Klitgaard, Kirstine; Boye, Mette; Capion, Nynne

    2008-01-01

    The etiopathogenesis of the skin disease digital dermatitis (DD), an important cause of lameness in cattle, remains uncertain. Microscopically, the disease appears to be polymicrobial, with spirochetes as the predominant bacteria. The objective of this study was to identify the main part of the b......The etiopathogenesis of the skin disease digital dermatitis (DD), an important cause of lameness in cattle, remains uncertain. Microscopically, the disease appears to be polymicrobial, with spirochetes as the predominant bacteria. The objective of this study was to identify the main part...... of the bacteria involved in DD lesions of cattle by using culture-independent molecular methods. Ten different phylotypes of Treponema were identified either by 16S rRNA gene sequencing of bacteria from DD lesions or by fluorescence in situ hybridization (FISH) analysis using phylotype-specific 16S r...... that Treponema accounted for more than 90% of the total bacterial population in the biopsy specimens. These data strongly suggest that a group of apparently symbiotic Treponema species are involved as primary bacterial pathogens in DD....

  2. Coumarinyl(thienyl)thiazoles: novel photochromes with modulated fluorescence.

    Science.gov (United States)

    Traven, Valery F; Bochkov, Andrei Y; Krayushkin, Mikhail M; Yarovenko, Vladimir N; Nabatov, Boris V; Dolotov, Sergei M; Barachevsky, Valery A; Beletskaya, Irina P

    2008-03-20

    Novel photochromic 5-(3'-coumarinyl)-4-(3''-thienyl)thiazoles have been synthesized. These compounds display intensive fluorescence emission in the open form A, which is modulated by light. Fluorescence intensity decreases significantly upon irradiation of A with UV-light (lambdaB. Irradiation of B with visible light (lambda>470 nm) promotes its opening and the recovering of fluorescence. Novel dihetarylethenes undergo photochromic modulation of fluorescence both in solution and in polymeric matrices.

  3. Effect of Different Fixatives and Passage Number on Fluorescence Intensity of pDsRed2-Mito Plasmid%固定剂和传代次数对线粒体荧光质粒 pDsRed2-Mito荧光强度的影响

    Institute of Scientific and Technical Information of China (English)

    张丽丽; 耿慧霞; 石贞玉; 邓锦波; 王来

    2016-01-01

    采用标记线粒体的荧光质粒 pDsRed2-Mito 转染 PC12细胞,分别以95%乙醇、甲醇、丙酮和4%多聚甲醛作为固定剂对筛选后的细胞进行固定,最后在激光共聚焦显微镜下观察不同固定剂对荧光质粒荧光强度的影响。结果表明,95%乙醇或甲醇作为固定剂固定细胞后易导致荧光猝灭,而丙酮或4%多聚甲醛固定细胞后,对质粒荧光强度没有显著影响。不同传代次数对 pDsRed2-Mito 质粒的荧光强度也没有显著影响。因此,在选择 pDsRed2-Mito 质粒标记线粒体时,不用考虑传代次数的影响,宜选择丙酮或4%多聚甲醛作为固定剂才能最大化的保持质粒的荧光强度。%In this paper,the pDsRed2-Mito plasmid,which specifically marks the mitochondrial morphology and structure in cells,was transfected to PC12 cells,and fixed by 95% ethanol,methyl alcohol,acetone and 4%paraformaldehyde respectively.The effect of different fixatives on fluorescence intensity of the plasmid was observed with laser scanning confocal microscope.The results indicate that the fluorescence intensity of pDsRed2-Mito would quench easily when the fixative was 95% ethanol or methyl alcohol,while the introduction of the acetone or 4%paraformaldehyde has no remarkable effect on its fluorescence intensity.Furthermore,the times of passages has also no significant impact on the fluorescence intensity of pDsRed2-Mito.In this way,with pDsRed2-Mito to label mitochondria,it is a good choose to use acetone and 4% paraformaldehyde as a fixative to maintain fluorescence intensity.

  4. Parent-child attitude congruence on type and intensity of physical activity: Testing multiple mediators of sedentary behavior in older children

    Science.gov (United States)

    This study examined parent–child attitudes on value of specific types and intensities of physical activity, which may explain gender differences in child activity, and evaluated physical activity as a mechanism to reduce time spent in sedentary behaviors. A community sample of 681 parents and 433 ch...

  5. Analysis of quantum dot fluorescence stability in primary blood mononuclear cells.

    Science.gov (United States)

    Summers, Huw D; Holton, Mark D; Rees, Paul; Williams, Paul M; Thornton, Catherine A

    2010-10-01

    A quantitative assessment of fluorescence signal generation and persistence in blood cells, measured at multiple points over a time course, is presented. Quantum dots (QDs) are inorganic fluorophores that are photostable and nonmetabolized and so can provide quantitative measures of cell biology over multiple cell generations. However, if the potential of these nanoparticles for long-term reporting is to be realized, an understanding of the stability of their fluorescence in living cells is essential. CdTe/ZnS and CdSe/ZnS core/shell dots with peak emission wavelengths of 705 nm and 585 nm, respectively, were loaded, via endocytosis into mononuclear cells extracted from primary blood and flow cytometry used to measure the average fluorescence intensity per cell within populations >10⁴. Time-based study showed a saturation-limited uptake of QDs with a characteristic time of 20 min and a maximum fluorescence signal that is linearly proportional to dot solution concentration. The fluorescence signal decreases after attachment and internalization within cells and is accurately described by a biexponential decay with a rapid initial decay followed by a much slower signal loss with characteristic times of 435 and 7,000 min respectively. Comparison with control samples indicates that interaction with the culture media is a major contributory factor to the initial signal decay. These results provide phenomenological descriptions of the evolving QD fluorescence within live cells with associated analytical equations that allow quantitative assessment of QD-based assays.

  6. Laser-induced fluorescence: quantitative analysis of atherosclerotic plaque chemical content in human aorta

    Science.gov (United States)

    Dai, Erbin; Wishart, David; Khoury, Samir; Kay, Cyril M.; Jugdutt, Bodh I.; Tulip, John; Lucas, Alexandra

    1996-05-01

    We have been studying laser-induced fluorescence as a technique for identification of selected changes in the chemical composition of atherosclerotic plaque. Formulae for quantification of chemical changes have been developed based upon analysis of fluorescence emission spectra using multiple regression analysis and the principal of least squares. The intima of human aortic necropsy specimens was injected with chemical compounds present in atherosclerotic plaque. Spectra recorded after injection of selected chemical components found in plaque (collagen I, III, IV, elastin and cholesterol) at varying concentrations (0.01 - 1.0 mg) were compared with saline injection. A single fiber system was used for both fluorescence excitation (XeCl excimer laser, 308 nm, 1.5 - 2.0 mJ/ pulse, 5 Hz) and fluorescence emission detection. Average spectra for each chemical have been developed and the wavelengths of peak emission intensity identified. Curve fitting analysis as well as multiple regression analysis were used to develop formulae for assessment of chemical content. Distinctive identifying average curves were established for each chemical. Excellent correlations were identified for collagen I, III, and IV, elastin, and cholesterol (R2 equals 0.92 6- 0.997). Conclusions: (1) Fluorescence spectra of human aortas were significantly altered by collagen I, collagen III, elastin and cholesterol. (2) Fluorescence spectroscopic analysis may allow quantitative assessment of atherosclerotic plaque chemical content in situ.

  7. Fluorescent microspheres

    Science.gov (United States)

    Rembaum, A.

    1978-01-01

    Latex particles with attached antibodies have potential biochemical and environmental applications. Human red blood cells and lymphocytes have been labeled with fluorescent microspheres by either direct or indirect immunological technique. Immunolatex spheres can also be used for detecting and localizing specific cell surface receptors. Hormones and toxins may also be bondable.

  8. Delayed fluorescence in photosynthesis.

    Science.gov (United States)

    Goltsev, Vasilij; Zaharieva, Ivelina; Chernev, Petko; Strasser, Reto J

    2009-01-01

    Photosynthesis is a very efficient photochemical process. Nevertheless, plants emit some of the absorbed energy as light quanta. This luminescence is emitted, predominantly, by excited chlorophyll a molecules in the light-harvesting antenna, associated with Photosystem II (PS II) reaction centers. The emission that occurs before the utilization of the excitation energy in the primary photochemical reaction is called prompt fluorescence. Light emission can also be observed from repopulated excited chlorophylls as a result of recombination of the charge pairs. In this case, some time-dependent redox reactions occur before the excitation of the chlorophyll. This delays the light emission and provides the name for this phenomenon-delayed fluorescence (DF), or delayed light emission (DLE). The DF intensity is a decreasing polyphasic function of the time after illumination, which reflects the kinetics of electron transport reactions both on the (electron) donor and the (electron) acceptor sides of PS II. Two main experimental approaches are used for DF measurements: (a) recording of the DF decay in the dark after a single turnover flash or after continuous light excitation and (b) recording of the DF intensity during light adaptation of the photosynthesizing samples (induction curves), following a period of darkness. In this paper we review historical data on DF research and recent advances in the understanding of the relation between the delayed fluorescence and specific reactions in PS II. An experimental method for simultaneous recording of the induction transients of prompt and delayed chlorophyll fluorescence and decay curves of DF in the millisecond time domain is discussed.

  9. Synthesis of novel fluorescent probe Tb(III)-7-carboxymethoxy-4-methylcoumarin complex for sensing of DNA

    Energy Technology Data Exchange (ETDEWEB)

    Hussein, Belal H.M., E-mail: belalhussein102@yahoo.com [Department of Chemistry, Faculty of Science, Suez Canal University, Ismailia (Egypt); Azab, Hassan A. [Department of Chemistry, Faculty of Science, Suez Canal University, Ismailia (Egypt); Fathalla, Walid [Department of Mathematical and Physical Sciences, Faculty of Engineering, Port-Said University, Port-Said (Egypt); Ali, Sherin A.M. [Department of Mathematical and Physical Sciences, Faculty of Engineering, Suez Canal University, Ismailia (Egypt)

    2013-02-15

    New fluorescent probe Tb(III) (7-carboxymethoxy-4-methylcoumarin)2(SCN) (C2H5OH)(H2O) was synthesized and characterized by spectroscopy and thermal analysis. The absorption and fluorescence spectra of 7-carboxymethoxy-4-methylcoumarin (CMMC) and Tb(III)-CMMC complex have been measured in different solvents. The interactions of Tb(III)-CMMC complex with calf thymus nucleic acid (CT-DNA) have been investigated using steady state fluorescence measurements. The changes in the fluorescence intensity have been used for the quantitative determination of DNA with LOD of 3.45 ng in methanol-water (9:1, v/v). The association constants of DNA with Tb(III)-CMMC complex was found to be 2.62 Multiplication-Sign 1010 M{sup -1}. - Highlights: Black-Right-Pointing-Pointer New fluorescent probe Terbium (III)-7-carboxy methoxy-4-methylcoumarin complex has been synthesized and characterized. Black-Right-Pointing-Pointer FTIR spectrum of Tb(III)-complex shows a characteristic band for thiocyanate group. Black-Right-Pointing-Pointer DNA interaction with Terbium (III)-7-carboxy methoxy-4-methylcoumarin has been studied by fluorescence techniques. Black-Right-Pointing-Pointer The change in the fluorescence intensity has been used for the quantitative determination of DNA. Black-Right-Pointing-Pointer The result was better than most of the well-known methods including the ethidium bromide method.

  10. Autologous/reduced-intensity allogeneic stem cell transplantation vs autologous transplantation in multiple myeloma: long-term results of the EBMT-NMAM2000 study

    NARCIS (Netherlands)

    Gahrton, G.; Iacobelli, S.; Bjorkstrand, B.; Hegenbart, U.; Gruber, A.; Greinix, H.; Volin, L.; Narni, F.; Carella, A.M.; Beksac, M.; Bosi, A.; Milone, G.; Corradini, P.; Schonland, S.; Friberg, K.; Biezen, A. van; Goldschmidt, H.; Witte, T.J.M. de; Morris, C.; Niederwieser, D.; Garderet, L.; Kroger, N.

    2013-01-01

    Long-term follow-up of prospective studies comparing allogeneic transplantation to autologous transplantation in multiple myeloma is few and controversial. This is an update at a median follow-up of 96 months of the European Group for Blood and Marrow Transplantation Non-Myeloablative Allogeneic ste

  11. Improved Stress Intensity Solutions Developed for the Multiple Site Damage Scenario: Two Unequal Through Cracks on Either Side of an Open Hole, Multiple Through Cracks, and Through Cracks Approaching an Open Hole

    Science.gov (United States)

    2014-10-01

    so the stress intensity factor at each crack tip is a function of the length of both cracks. The handbook solution by Murakami [3], was reported to...G.R., "The Stress Analysis of Cracks Handbook," Second Edition, Paris Productions, Inc., St. Louis, MO, 1985 3. Murakami , Y., et al., "Stress

  12. Control of excitation in the fluorescence microscope.

    Science.gov (United States)

    Lea, D J; Ward, D J

    1979-01-01

    In fluorescence microscopy image brightness and contrast and the rate of fading depend upon the intensity of illumination of the specimen. An iris diaphragm or neutral density filters may be used to reduce fluorescence excitation. Also the excitation bandwidth may be varied by using a broad band exciter filter with a set of interchangeable yellow glass filters at the lamphouse.

  13. Studying Photosynthesis by Measuring Fluorescence

    Science.gov (United States)

    Sanchez, Jose Francisco; Quiles, Maria Jose

    2006-01-01

    This paper describes an easy experiment to study the absorption and action spectrum of photosynthesis, as well as the inhibition by heat, high light intensity and the presence of the herbicide 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) on the photosynthetic process. The method involves measuring the chlorophyll fluorescence emitted by intact…

  14. Quantum dot imaging in the second near-infrared optical window: studies on reflectance fluorescence imaging depths by effective fluence rate and multiple image acquisition.

    Science.gov (United States)

    Jung, Yebin; Jeong, Sanghwa; Nayoun, Won; Ahn, Boeun; Kwag, Jungheon; Geol Kim, Sang; Kim, Sungjee

    2015-04-01

    Quantum dot (QD) imaging capability was investigated by the imaging depth at a near-infrared second optical window (SOW; 1000 to 1400 nm) using time-modulated pulsed laser excitations to control the effective fluence rate. Various media, such as liquid phantoms, tissues, and in vivo small animals, were used and the imaging depths were compared with our predicted values. The QD imaging depth under excitation of continuous 20 mW/cm(2) laser was determined to be 10.3 mm for 2 wt%hemoglobin phantom medium and 5.85 mm for 1 wt% intralipid phantom, which were extended by more than two times on increasing the effective fluence rate to 2000 mW/cm(2). Bovine liver and porcine skin tissues also showed similar enhancement in the contrast-to-noise ratio (CNR) values. A QD sample was inserted into the abdomen of a mouse.With a higher effective fluence rate, the CNR increased more than twofold and the QD sample became clearly visualized, which was completely undetectable under continuous excitation.Multiple acquisitions of QD images and averaging process pixel by pixel were performed to overcome the thermal noise issue of the detector in SOW, which yielded significant enhancement in the imaging capability, showing up to a 1.5 times increase in the CNR.

  15. Nano-sensing of the orientation of fluorescing molecules with active coated nano-particles

    DEFF Research Database (Denmark)

    Arslanagic, Samel; Ziolkowski, Richard W.

    2015-01-01

    The potential of using active coated nano-particles to determine the orientation of fluorescing molecules is reported. By treating each fluorescing molecule as an electric Hertzian dipole, single and multiple fluorescing molecules emitting coherently and incoherently in various orientations...

  16. Quasi-real-time fluorescence imaging with lifetime dependent contrast

    Science.gov (United States)

    Jiang, Pei-Chi; Grundfest, Warren S.; Stafsudd, Oscar M.

    2011-08-01

    Conventional fluorescence lifetime imaging requires complicated algorithms to extract lifetimes of fluorophores and acquisition of multiple data points at progressively longer delay times to characterize tissues. To address diminishing signal-to-noise ratios at these progressively longer time delays, we report a time-resolved fluorescence imaging method, normalized fluorescence yield imaging that does not require the extraction of lifetimes. The concept is to extract the ``contrast'' instead of the lifetime value of the fluorophores by using simple mathematical algorithms. This process converts differences in decay times directly to different intensities. The technique was verified experimentally using a gated iCCD camera and an ultraviolet light-emitting diode light source. It was shown that this method can distinguish between chemical dyes (Fluorescein and Rhodamine-B) and biomedical samples, such as powders of elastin and collagen. Good contrast was obtained between fluorophores that varied by less than 6% in lifetime. Additionally, it was shown that long gate times up to 16 ns achieve good contrast depending upon the samples to be studied. These results support the feasibility of time-resolved fluorescence imaging without lifetime extraction, which has a potential clinical role in noninvasive real-time imaging.

  17. Nanostructured surfaces and detection instrumentation for photonic crystal enhanced fluorescence.

    Science.gov (United States)

    Chaudhery, Vikram; George, Sherine; Lu, Meng; Pokhriyal, Anusha; Cunningham, Brian T

    2013-04-26

    Photonic crystal (PC) surfaces have been demonstrated as a compelling platform for improving the sensitivity of surface-based fluorescent assays used in disease diagnostics and life science research. PCs can be engineered to support optical resonances at specific wavelengths at which strong electromagnetic fields are utilized to enhance the intensity of surface-bound fluorophore excitation. Meanwhile, the leaky resonant modes of PCs can be used to direct emitted photons within a narrow range of angles for more efficient collection by a fluorescence detection system. The multiplicative effects of enhanced excitation combined with enhanced photon extraction combine to provide improved signal-to-noise ratios for detection of fluorescent emitters, which in turn can be used to reduce the limits of detection of low concentration analytes, such as disease biomarker proteins. Fabrication of PCs using inexpensive manufacturing methods and materials that include replica molding on plastic, nano-imprint lithography on quartz substrates result in devices that are practical for single-use disposable applications. In this review, we will describe the motivation for implementing high-sensitivity fluorescence detection in the context of molecular diagnosis and gene expression analysis though the use of PC surfaces. Recent efforts to improve the design and fabrication of PCs and their associated detection instrumentation are summarized, including the use of PCs coupled with Fabry-Perot cavities and external cavity lasers.

  18. Nanostructured Surfaces and Detection Instrumentation for Photonic Crystal Enhanced Fluorescence

    Directory of Open Access Journals (Sweden)

    Brian T. Cunningham

    2013-04-01

    Full Text Available Photonic crystal (PC surfaces have been demonstrated as a compelling platform for improving the sensitivity of surface-based fluorescent assays used in disease diagnostics and life science research. PCs can be engineered to support optical resonances at specific wavelengths at which strong electromagnetic fields are utilized to enhance the intensity of surface-bound fluorophore excitation. Meanwhile, the leaky resonant modes of PCs can be used to direct emitted photons within a narrow range of angles for more efficient collection by a fluorescence detection system. The multiplicative effects of enhanced excitation combined with enhanced photon extraction combine to provide improved signal-to-noise ratios for detection of fluorescent emitters, which in turn can be used to reduce the limits of detection of low concentration analytes, such as disease biomarker proteins. Fabrication of PCs using inexpensive manufacturing methods and materials that include replica molding on plastic, nano-imprint lithography on quartz substrates result in devices that are practical for single-use disposable applications. In this review, we will describe the motivation for implementing high-sensitivity fluorescence detection in the context of molecular diagnosis and gene expression analysis though the use of PC surfaces. Recent efforts to improve the design and fabrication of PCs and their associated detection instrumentation are summarized, including the use of PCs coupled with Fabry-Perot cavities and external cavity lasers.

  19. Modular generation of fluorescent phycobiliproteins.

    Science.gov (United States)

    Wu, Xian-Jun; Chang, Kun; Luo, Juan; Zhou, Ming; Scheer, Hugo; Zhao, Kai-Hong

    2013-06-01

    Phycobiliproteins are brightly-fluorescent light-harvesting pigments for photosynthesis in cyanobacteria and red algae. They are also of interest as fluorescent biomarkers, but their heterologous generation in vivo has previously required multiple transformations. We report here a modular approach that requires only two DNA segments. The first codes for the apo-protein. The second codes for fusions capable of chromophore biosynthesis and its covalent attachment to the apo-protein; it contains the genes of heme oxygenase, a bilin reductase, and a chromophore lyase. Phycobiliproteins containing phycoerythrobilin (λ(fluor) ~ 560 nm), phycourobilin (λ(fluor) ~ 500 nm), phycocyanobilin (λ(fluor) ~ 630 nm) or phycoviolobilin (λ(fluor) ~ 580 nm) were obtained in high yield in E. coli. This approach facilitates chromophorylation studies of phycobiliproteins, as well as their use for fluorescence labeling based on their high fluorescence.

  20. Enhanced speed in fluorescence imaging using beat frequency multiplexing

    Science.gov (United States)

    Mikami, Hideharu; Kobayashi, Hirofumi; Wang, Yisen; Hamad, Syed; Ozeki, Yasuyuki; Goda, Keisuke

    2016-03-01

    Fluorescence imaging using radiofrequency-tagged emission (FIRE) is an emerging technique that enables higher imaging speed (namely, temporal resolution) in fluorescence microscopy compared to conventional fluorescence imaging techniques such as confocal microscopy and wide-field microscopy. It works based on the principle that it uses multiple intensity-modulated fields in an interferometric setup as excitation fields and applies frequency-division multiplexing to fluorescence signals. Unfortunately, despite its high potential, FIRE has limited imaging speed due to two practical limitations: signal bandwidth and signal detection efficiency. The signal bandwidth is limited by that of an acousto-optic deflector (AOD) employed in the setup, which is typically 100-200 MHz for the spectral range of fluorescence excitation (400-600 nm). The signal detection efficiency is limited by poor spatial mode-matching between two interfering fields to produce a modulated excitation field. Here we present a method to overcome these limitations and thus to achieve higher imaging speed than the prior version of FIRE. Our method achieves an increase in signal bandwidth by a factor of two and nearly optimal mode matching, which enables the imaging speed limited by the lifetime of the target fluorophore rather than the imaging system itself. The higher bandwidth and better signal detection efficiency work synergistically because higher bandwidth requires higher signal levels to avoid the contribution of shot noise and amplifier noise to the fluorescence signal. Due to its unprecedentedly high-speed performance, our method has a wide variety of applications in cancer detection, drug discovery, and regenerative medicine.

  1. IKZF1 expression is a prognostic marker in newly diagnosed standard-risk multiple myeloma treated with lenalidomide and intensive chemotherapy: a study of the German Myeloma Study Group (DSMM).

    Science.gov (United States)

    Krönke, J; Kuchenbauer, F; Kull, M; Teleanu, V; Bullinger, L; Bunjes, D; Greiner, A; Kolmus, S; Köpff, S; Schreder, M; Mügge, L-O; Straka, C; Engelhardt, M; Döhner, H; Einsele, H; Bassermann, F; Bargou, R; Knop, S; Langer, C

    2017-01-20

    Lenalidomide is an immunomodulatory compound with high clinical activity in multiple myeloma. Lenalidomide binding to the Cereblon (CRBN) E3 ubiquitin ligase results in targeted ubiquitination and degradation of the lymphoid transcription factors Ikaros (IKZF1) and Aiolos (IKZF3) leading to growth inhibition of multiple myeloma cells. Recently, Basigin (BSG) was identified as another protein regulated by CRBN that is involved in the activity of lenalidomide. Here, we analyzed the prognostic value of IKZF1, IKZF3, CRBN and BSG mRNA expression levels in pretreatment plasma cells from 60 patients with newly diagnosed multiple myeloma uniformly treated with lenalidomide in combination with intensive chemotherapy within a clinical trial. We found that IKZF1 mRNA expression levels are significantly associated with progression-free survival (PFS). Patients in the lowest quartile (Q1) of IKZF1 expression had a superior PFS compared with patients in the remaining quartiles (Q2-Q4; 3-year PFS of 86 vs 51%, P=0.01). This translated into a significant better overall survival (100 vs 74%, P=0.03). Subgroup analysis revealed a significant impact of IKZF1, IKZF3 and BSG expression levels on PFS in cytogenetically defined standard-risk but not high-risk patients. Our data suggest a prognostic role of IKZF1, IKZF3 and BSG expression levels in lenalidomide-treated multiple myeloma.Leukemia advance online publication, 20 January 2017; doi:10.1038/leu.2016.384.

  2. Increasing signal intensity within the dentate nucleus and globus pallidus on unenhanced T1W magnetic resonance images in patients with relapsing-remitting multiple sclerosis: correlation with cumulative dose of a macrocyclic gadolinium-based contrast agent, gadobutrol

    Energy Technology Data Exchange (ETDEWEB)

    Stojanov, Dragan A. [University of Nis, Faculty of Medicine, Nis (Serbia); Clinical Center Nis, Center for Radiology, Nis (Serbia); Aracki-Trenkic, Aleksandra [Clinical Center Nis, Center for Radiology, Nis (Serbia); Vojinovic, Slobodan; Ljubisavljevic, Srdjan [University of Nis, Faculty of Medicine, Nis (Serbia); Clinical Center Nis, Clinic for Neurology, Nis (Serbia); Benedeto-Stojanov, Daniela [University of Nis, Faculty of Medicine, Nis (Serbia)

    2016-03-15

    To evaluate correlation between cumulative dose of gadobutrol and signal intensity (SI) within dentate nucleus and globus pallidus on unenhanced T1-weighted images in patients with relapsing-remitting multiple sclerosis (RRMS). Dentate nucleus-to-pons and globus pallidus-to-thalamus SI ratios, and renal and liver functions, were evaluated after multiple intravenous administrations of 0.1 mmol/kg gadobutrol at 27, 96-98, and 168 weeks. We compared SI ratios based on the number of administrations, total amount of gadobutrol administered, and time between injections. Globus pallidus-to-thalamus (p = 0.025) and dentate nucleus-to-pons (p < 0.001) SI ratios increased after multiple gadobutrol administrations, correlated with the number of administrations (ρ = 0.263, p = 0.046, respectively) and depended on the length of administration (p = 0.017, p = 0.037, respectively). Patients receiving gadobutrol at 27 weeks showed the greatest increase in both SI ratios (p = 0.006; p = 0.014, respectively, versus 96-98 weeks). GGT increased at the end of the study (p = 0.004). In patients with RRMS, SI within the dentate nucleus and globus pallidus increased on unenhanced T1-weighted images after multiple gadobutrol injections. Administration of the same total amount of gadobutrol over a shorter period caused greater SI increase. (orig.)

  3. Human cytomegaloviruses expressing yellow fluorescent fusion proteins--characterization and use in antiviral screening.

    Science.gov (United States)

    Straschewski, Sarah; Warmer, Martin; Frascaroli, Giada; Hohenberg, Heinrich; Mertens, Thomas; Winkler, Michael

    2010-02-11

    Recombinant viruses labelled with fluorescent proteins are useful tools in molecular virology with multiple applications (e.g., studies on intracellular trafficking, protein localization, or gene activity). We generated by homologous recombination three recombinant cytomegaloviruses carrying the enhanced yellow fluorescent protein (EYFP) fused with the viral proteins IE-2, ppUL32 (pp150), and ppUL83 (pp65). In growth kinetics, the three viruses behaved all like wild type, even at low multiplicity of infection (MOI). The expression of all three fusion proteins was detected, and their respective localizations were the same as for the unmodified proteins in wild-type virus-infected cells. We established the in vivo measurement of fluorescence intensity and used the recombinant viruses to measure inhibition of viral replication by neutralizing antibodies or antiviral substances. The use of these viruses in a pilot screen based on fluorescence intensity and high-content analysis identified cellular kinase inhibitors that block viral replication. In summary, these viruses with individually EYFP-tagged proteins will be useful to study antiviral substances and the dynamics of viral infection in cell culture.

  4. Human cytomegaloviruses expressing yellow fluorescent fusion proteins--characterization and use in antiviral screening.

    Directory of Open Access Journals (Sweden)

    Sarah Straschewski

    Full Text Available Recombinant viruses labelled with fluorescent proteins are useful tools in molecular virology with multiple applications (e.g., studies on intracellular trafficking, protein localization, or gene activity. We generated by homologous recombination three recombinant cytomegaloviruses carrying the enhanced yellow fluorescent protein (EYFP fused with the viral proteins IE-2, ppUL32 (pp150, and ppUL83 (pp65. In growth kinetics, the three viruses behaved all like wild type, even at low multiplicity of infection (MOI. The expression of all three fusion proteins was detected, and their respective localizations were the same as for the unmodified proteins in wild-type virus-infected cells. We established the in vivo measurement of fluorescence intensity and used the recombinant viruses to measure inhibition of viral replication by neutralizing antibodies or antiviral substances. The use of these viruses in a pilot screen based on fluorescence intensity and high-content analysis identified cellular kinase inhibitors that block viral replication. In summary, these viruses with individually EYFP-tagged proteins will be useful to study antiviral substances and the dynamics of viral infection in cell culture.

  5. Data-intensive science

    CERN Document Server

    Critchlow, Terence

    2013-01-01

    Data-intensive science has the potential to transform scientific research and quickly translate scientific progress into complete solutions, policies, and economic success. But this collaborative science is still lacking the effective access and exchange of knowledge among scientists, researchers, and policy makers across a range of disciplines. Bringing together leaders from multiple scientific disciplines, Data-Intensive Science shows how a comprehensive integration of various techniques and technological advances can effectively harness the vast amount of data being generated and significan

  6. Detection of Counterfeit Tequila by Fluorescence Spectroscopy

    OpenAIRE

    José Manuel de la Rosa Vázquez; Diego Adrián Fabila-Bustos; Luis Felipe de Jesús Quintanar-Hernández; Alma Valor; Suren Stolik

    2015-01-01

    An ultraviolet (UV) light induced fluorescence study to discriminate fake tequila from genuine ones is presented. A portable homemade system based on four light emitting diodes (LEDs) from 255 to 405 nm and a miniature spectrometer was used. It has been shown that unlike fake and silver tequila, which produce weak fluorescence signal, genuine mixed, rested, and aged tequilas show high fluorescence emission in the range from 400 to 750 nm. The fluorescence intensity grows with aging in 100% ag...

  7. Differentiation of cancerous and normal brain tissue using label free fluorescence and Stokes shift spectroscopy

    Science.gov (United States)

    Zhou, Yan; Wang, Leana; Liu, Cheng-hui; He, Yong; Yu, Xinguang; Cheng, Gangge; Wang, Peng; Shu, Cheng; Alfano, Robert R.

    2016-03-01

    In this report, optical biopsy was applied to diagnose human brain cancer in vitro for the identification of brain cancer from normal tissues by native fluorescence and Stokes shift spectra (SSS). 77 brain specimens including three types of human brain tissues (normal, glioma and brain metastasis of lung cancers) were studied. In order to observe spectral changes of fluorophores via fluorescence, the selected excitation wavelength of UV at 300 and 340 nm for emission spectra and a different Stokes Shift spectra with intervals Δλ = 40 nm were measured. The fluorescence spectra and SSS from multiple key native molecular markers, such as tryptophan, collagen, NADH, alanine, ceroid and lipofuscin were observed in normal and diseased brain tissues. Two diagnostic criteria were established based on the ratios of the peak intensities and peak position in both fluorescence and SSS spectra. It was observed that the ratio of the spectral peak intensity of tryptophan (340 nm) to NADH (440 nm) increased in glioma, meningioma (benign), malignant meninges tumor, and brain metastasis of lung cancer tissues in comparison with normal tissues. The ratio of the SS spectral peak (Δλ = 40 nm) intensities from 292 nm to 366 nm had risen similarly in all grades of tumors.

  8. Measurement of event-by-event transverse momentum and multiplicity fluctuations using strongly intensive measures $\\Delta[P_T, N]$ and $\\Sigma[P_T, N]$ in nucleus-nucleus collisions at the CERN Super Proton Synchrotron

    CERN Document Server

    Anticic, T; Bartke, J; Beck, H; Betev, L; Bialkowska, H; Blume, C; Boimska, B; Book, J; Botje, M; Buncic, P; Christakoglou, P; Chung, P; Chvala, O; Cramer, J; Eckardt, V; Fodor, Z; Foka, P; Friese, V; Gazdzicki, M; Grebieszkow, K; Hohne, C; Kadija, K; Karev, A; Kolesnikov, V; Kowalski, M; Kresan, D; Laszlo, A; Lacey, R; van Leeuwen, M; Mackowiak-Pawlowska, M; Makariev, M; Malakhov, A; Melkumov, G; Mitrovski, M; Mrowczynski, S; Palla, G; Panagiotou, A; Pluta, J; Prindle, D; Puhlhofer, F; Renfordt, R; Roland, C; Roland, G; Rybczynski, M; Rybicki, A; Sandoval, A; Rustamov, A; Schmitz, N; Schuster, T; Seyboth, P; Sikler, F; Skrzypczak, E; Slodkowski, M; Stefanek, G; Stock, R; Strobele, H; Susa, T; Szuba, M; Varga, D; Vassiliou, M; Veres, G; Vesztergombi, G; Vranic, D; Wlodarczyk, Z; Wojtaszek-Szwarc, A

    2015-01-01

    Results from the NA49 experiment at the CERN SPS are presented on event-by-event transverse momentum and multiplicity fluctuations of charged particles, produced at forward rapidities in central Pb+Pb interactions at beam momenta 20$A$, 30$A$, 40$A$, 80$A$, and 158$A$ GeV/c, as well as in systems of different size ($p+p$, C+C, Si+Si, and Pb+Pb) at 158$A$ GeV/c. This publication extends the previous NA49 measurements of the strongly intensive measure $\\Phi_{p_T}$ by a study of the recently proposed strongly intensive measures of fluctuations $\\Delta[P_T, N]$ and $\\Sigma[P_T, N]$. In the explored kinematic region transverse momentum and multiplicity fluctuations show no significant energy dependence in the SPS energy range. However, a remarkable system size dependence is observed for both $\\Delta[P_T, N]$ and $\\Sigma[P_T, N]$, with the largest values measured in peripheral Pb+Pb interactions. The results are compared with NA61/SHINE measurements in $p+p$ collisions, as well as with predictions of the UrQMD and ...

  9. Multichromophoric sugar for fluorescence photoswitching

    Directory of Open Access Journals (Sweden)

    Stéphane Maisonneuve

    2014-06-01

    Full Text Available A multichromophoric glucopyranoside 2 bearing three dicyanomethylenepyran (DCM fluorophores and one diarylethene (DAE photochrome has been prepared by Cu(I-catalyzed alkyne–azide cycloaddition reaction. The fluorescence of 2 was switched off upon UV irradiation, in proportion with the open to closed form (OF to CF conversion extent of the DAE moiety. A nearly 100% Förster-type resonance energy transfer (FRET from all three DCM moieties to a single DAE (in its CF moiety was achieved. Upon visible irradiation, the initial fluorescence intensity was recovered. The observed photoswiching is reversible, with excellent photo resistance.

  10. Fluorescent Approaches to High Throughput Crystallography

    Science.gov (United States)

    Pusey, Marc L.; Forsythe, Elizabeth; Achari, Aniruddha

    2006-01-01

    We have shown that by covalently modifying a subpopulation, less than or equal to 1%, of a macromolecule with a fluorescent probe, the labeled material will add to a growing crystal as a microheterogeneous growth unit. Labeling procedures can be readily incorporated into the final stages of purification, and the presence of the probe at low concentrations does not affect the X-ray data quality or the crystallization behavior. The presence of the trace fluorescent label gives a number of advantages when used with high throughput crystallizations. The covalently attached probe will concentrate in the crystal relative to the solution, and under fluorescent illumination crystals show up as bright objects against a dark background. Non-protein structures, such as salt crystals, will not incorporate the probe and will not show up under fluorescent illumination. Brightly fluorescent crystals are readily found against less bright precipitated phases, which under white light illumination may obscure the crystals. Automated image analysis to find crystals should be greatly facilitated, without having to first define crystallization drop boundaries as the protein or protein structures is all that shows up. Fluorescence intensity is a faster search parameter, whether visually or by automated methods, than looking for crystalline features. We are now testing the use of high fluorescence intensity regions, in the absence of clear crystalline features or "hits", as a means for determining potential lead conditions. A working hypothesis is that kinetics leading to non-structured phases may overwhelm and trap more slowly formed ordered assemblies, which subsequently show up as regions of brighter fluorescence intensity. Preliminary experiments with test proteins have resulted in the extraction of a number of crystallization conditions from screening outcomes based solely on the presence of bright fluorescent regions. Subsequent experiments will test this approach using a wider

  11. Timing and Operating Mode Design for Time-Gated Fluorescence Lifetime Imaging Microscopy

    OpenAIRE

    Chao Liu; Xinwei Wang; Yan Zhou; Yuliang Liu

    2013-01-01

    Steady-state fluorence imaging and time-resolved fluorescence imaging are two important areas in fluorescence imaging research. Fluorescence lifetime imaging is an absolute measurement method which is independent of excitation laser intensity, fluorophore concentration, and photobleaching compared to fluorescence intensity imaging techniques. Time-gated fluorescence lifetime imaging microscopy (FLIM) can provide high resolution and high imaging frame during mature FLIM methods. An abstract ti...

  12. Spectroscopic quantification of 5-hydroxymethylcytosine in genomic DNA using boric acid-functionalized nano-microsphere fluorescent probes.

    Science.gov (United States)

    Chen, Hua-Yan; Wei, Jing-Ru; Pan, Jiong-Xiu; Zhang, Wei; Dang, Fu-Quan; Zhang, Zhi-Qi; Zhang, Jing

    2017-05-15

    5-hydroxymethylcytosine (5hmC) is the sixth base of DNA. It is involved in active DNA demethylation and can be a marker of diseases such as cancer. In this study, we developed a simple and sensitive 2-(4-boronophenyl)quinoline-4-carboxylic acid modified poly (glycidyl methacrylate (PBAQA-PGMA) fluorescent probe to detect the 5hmC content of genomic DNA based on T4 β-glucosyltransferase-catalyzed glucosylation of 5hmC. The fluorescence-enhanced intensity recorded from the DNA sample was proportional to its 5-hydroxymethylcytosine content and could be quantified by fluorescence spectrophotometry. The developed probe showed good detection sensitivity and selectivity and a good linear relationship between the fluorescence intensity and the concentration of 5 hmC within a 0-100nM range. Compared with other fluorescence detection methods, this method not only could determine trace amounts of 5 hmC from genomic DNA but also could eliminate the interference of fluorescent dyes and the need for purification. It also could avoid multiple labeling. Because the PBAQA-PGMA probe could enrich the content of glycosyl-5-hydroxymethyl-2-deoxycytidine from a complex ground substance, it will broaden the linear detection range and improve sensitivity. The limit of detection was calculated to be 0.167nM after enrichment. Furthermore, the method was successfully used to detect 5-hydroxymethylcytosine from mouse tissues. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. SU-E-T-450: Dosimetric Impact of Rotational Error On Multiple-Target Intensity-Modulated Radiosurgery (IMRS) with Single-Isocenter

    Energy Technology Data Exchange (ETDEWEB)

    Jang, S; Huq, M [University of Pittsburgh Medical Center, Pittsburgh, PA (United States)

    2014-06-01

    Purpose: Evaluating the dosimetric-impact on multiple-targets placed away from the isocenter-target with varying rotational-error introduced by initial setup uncertainty and/or intrafractional-movement Methods: CyberKnife-Phantom was scanned with the Intracranial SRS-protocol of 1.25mm slice-thickness and the multiple-targets(GTV) of 1mm and 10mm in diameter were contoured on the Eclipse. PTV for distal-target only was drawn with 1mm expansion around the GTV to find out how much margin is needed to compensate for the rotational-error. The separation between the isocenter-target and distal-target was varied from 3cm to 7cm. RapidArc-based IMRS plans of 16Gy single-fraction were generated with five non-coplanar arcs by using Varian TrueBeam-STx equipped with high resolution MLC leaves of 2.5mm at center and with dose-rate of 1400MU/min at 6MV for flatteringfilter- free(FFF). An identical CT image with intentionally introduced 1° rotational-error was registered with the planning CT image, and the isodose distribution and Dose-Volume-Histogram(DVH) were compared with the original plans. Additionally, the dosimetric-impact of rotational error was evaluated with that of 6X photon energy which was generated with the same target-coverage. Results: For the 1mm-target with 6X-FFF, PTV-coverage(D100) of the distal-target with 1° rotational-error decreased from 1.00 to 0.35 as the separation between isocenter-target and distal-target increased from 3cm to 7cm. However, GTV-coverage(D100) was 1.0 except that of 7cm-separation(0.55), which resulted from the 1mm-margin around the distal-target. For 6X photon, GTV-coverage remained at 1.0 regardless of the separation of targets, showing that the dosimetric-impact of rotational error depends on the degree of rotational-error, separation of targets, and dose distribution around targets. For 10mm-target, PTV-coverage of distaltarget located 3cm-away was better than that of 1mm-target(0.93 versus 0.7) and GTV-coverage was 1

  14. Fluorescence lifetime plate reader: resolution and precision meet high-throughput.

    Science.gov (United States)

    Petersen, Karl J; Peterson, Kurt C; Muretta, Joseph M; Higgins, Sutton E; Gillispie, Gregory D; Thomas, David D

    2014-11-01

    We describe a nanosecond time-resolved fluorescence spectrometer that acquires fluorescence decay waveforms from each well of a 384-well microplate in 3 min with signal-to-noise exceeding 400 using direct waveform recording. The instrument combines high-energy pulsed laser sources (5-10 kHz repetition rate) with a photomultiplier and high-speed digitizer (1 GHz) to record a fluorescence decay waveform after each pulse. Waveforms acquired from rhodamine or 5-((2-aminoethyl)amino) naphthalene-1-sulfonic acid dyes in a 384-well plate gave lifetime measurements 5- to 25-fold more precise than the simultaneous intensity measurements. Lifetimes as short as 0.04 ns were acquired by interleaving with an effective sample rate of 5 GHz. Lifetime measurements resolved mixtures of single-exponential dyes with better than 1% accuracy. The fluorescence lifetime plate reader enables multiple-well fluorescence lifetime measurements with an acquisition time of 0.5 s per well, suitable for high-throughput fluorescence lifetime screening applications.

  15. Detection of Counterfeit Tequila by Fluorescence Spectroscopy

    Directory of Open Access Journals (Sweden)

    José Manuel de la Rosa Vázquez

    2015-01-01

    Full Text Available An ultraviolet (UV light induced fluorescence study to discriminate fake tequila from genuine ones is presented. A portable homemade system based on four light emitting diodes (LEDs from 255 to 405 nm and a miniature spectrometer was used. It has been shown that unlike fake and silver tequila, which produce weak fluorescence signal, genuine mixed, rested, and aged tequilas show high fluorescence emission in the range from 400 to 750 nm. The fluorescence intensity grows with aging in 100% agave tequila. Such fluorescence differences can even be observed with naked eyes. The presented results demonstrate that the fluorescence measurement could be a good method to detect counterfeit tequila.

  16. A label-free, fluorescence based assay for microarray

    Science.gov (United States)

    Niu, Sanjun

    intensity of excitation light. The fluorescence contrast is used to quantify the amount of probe-target hybridization. A mathematical model that considers multiple reflections and scattering is developed to explain the mechanism of the fluorescence contrast which depends on the thickness of the PS film. Scattering is the dominant factor that contributes to the contrast. The potential of this assay to detect single nucleotide polymorphism is also tested.

  17. Treatment Plan Technique and Quality for Single-Isocenter Stereotactic Ablative Radiotherapy of Multiple Lung Lesions with Volumetric-Modulated Arc Therapy or Intensity-Modulated Radiosurgery.

    Science.gov (United States)

    Quan, Kimmen; Xu, Karen M; Lalonde, Ron; Horne, Zachary D; Bernard, Mark E; McCoy, Chuck; Clump, David A; Burton, Steven A; Heron, Dwight E

    2015-01-01

    The aim of this study is to provide a practical approach to the planning technique and evaluation of plan quality for the multi-lesion, single-isocenter stereotactic ablative radiotherapy (SABR) of the lung. Eleven patients with two or more lung lesions underwent single-isocenter volumetric-modulated arc therapy (VMAT) radiosurgery or IMRS. All plans were normalized to the target maximum dose. For each plan, all targets were treated to the same dose. Plan conformity and dose gradient were maximized with dose-control tuning structures surrounding targets. For comparison, multi-isocenter plans were retrospectively created for four patients. Conformity index (CI), homogeneity index (HI), gradient index (GI), and gradient distance (GD) were calculated for each plan. V5, V10, and V20 of the lung and organs at risk (OARs) were collected. Treatment time and total monitor units (MUs) were also recorded. One patient had four lesions and the remainder had two lesions. Six patients received VMAT and five patients received intensity-modulated radiosurgery (IMRS). For those treated with VMAT, two patients received 3-arc VMAT and four received 2-arc VMAT. For those treated with IMRS, two patients were treated with 10 and 11 beams, respectively, and the rest received 12 beams. Prescription doses ranged from 30 to 54 Gy in three to five fractions. The median prescribed isodose line was 84% (range: 80-86%). The median maximum dose was 57.1 Gy (range: 35.7-65.1 Gy). The mean combined PTV was 49.57 cm(3) (range: 14.90-87.38 cm(3)). For single-isocenter plans, the median CI was 1.15 (range: 0.97-1.53). The median HI was 1.19 (range: 1.16-1.28). The median GI was 4.60 (range: 4.16-7.37). The median maximum radiation dose (Dmax) to total lung was 55.6 Gy (range: 35.7-62.0 Gy). The median mean radiation dose to the lung (Dmean) was 4.2 Gy (range: 1.1-9.3 Gy). The median lung V5 was 18.7% (range: 3.8-41.3%). There was no significant difference in CI, HI, GI, GD, V5, V10

  18. 肺炎克雷伯菌致多发性肝脓肿患者的加强护理%The intensive care to patients with multiple liver abscess caused by Klebsiella pneumonia

    Institute of Scientific and Technical Information of China (English)

    顾朝丽; 单荣芳; 刘颖; 黄金兰; 蒋雅琼

    2011-01-01

    Objective To conclude the successful rescue and intensive care to patients with multiple liver abscess caused by Klebsiella pneumonia. Methods The rescuing coordination to two critically ill patients, the nursing of liver puncture and tube indwelling, incision and drainage were summarized. The characteristics of the infection of Klebsiella pneumonia and the prevention of cross infection, the nursing of high fever, medicine taking, basic nursing, nutrition support, psychological nursing, discharge instructions and so on were introduced as well. Results After intensive care, two patients were discharged from hospital after recovery. Conclusion Klebsiella pneumonia is multidrug resistant, which can cause multiple liver abscess. Patients with multiple liver abscess are seriously sick and it takes a long time to cure them. The key to a successful rescue includes intensive care, dealing with the life - threatened symptoms and physical signs, early using anti - biotic drugs, cutting open the pyogenic liver abscess and leading the liquid out effectively and preventing the cross infection.%目的 总结肺炎克雷伯菌致多发性肝脓肿患者的成功抢救与加强护理.方法 介绍2例危重患者的抢救配合、肝脓肿穿刺置管与手术切开冲洗引流护理、肺炎克雷伯菌感染特点与预防交叉感染、高热护理、用药护理、基础护理、营养支持、心理护理、出院指导等.结果 2例患者经加强护理,完全康复出院.结论 肺炎克雷伯菌是常见的多重耐药菌,其引起的多发性肝脓肿患者病情危重、病程长,加强监护、积极处理危及生命的症状和体征、尽早使用敏感抗菌药物、有效的脓肿切开引流、预防交义感染是抢救成功的关键.

  19. CdSe/ZnS quantum dot fluorescence spectra shape-based thermometry via neural network reconstruction

    Energy Technology Data Exchange (ETDEWEB)

    Munro, Troy [Multiscale Thermal-Physics Lab, Department of Mechanical and Aerospace Engineering, Utah State University, 4130 Old Main Hill, Logan, Utah 84322 (United States); Laboratory of Soft Matter and Biophysics, Department of Physics and Astronomy, KU Leuven, Celestijnenlaan 200D, B-3001 Heverlee (Belgium); Liu, Liwang; Glorieux, Christ [Laboratory of Soft Matter and Biophysics, Department of Physics and Astronomy, KU Leuven, Celestijnenlaan 200D, B-3001 Heverlee (Belgium); Ban, Heng [Multiscale Thermal-Physics Lab, Department of Mechanical and Aerospace Engineering, Utah State University, 4130 Old Main Hill, Logan, Utah 84322 (United States)

    2016-06-07

    As a system of interest gets small, due to the influence of the sensor mass and heat leaks through the sensor contacts, thermal characterization by means of contact temperature measurements becomes cumbersome. Non-contact temperature measurement offers a suitable alternative, provided a reliable relationship between the temperature and the detected signal is available. In this work, exploiting the temperature dependence of their fluorescence spectrum, the use of quantum dots as thermomarkers on the surface of a fiber of interest is demonstrated. The performance is assessed of a series of neural networks that use different spectral shape characteristics as inputs (peak-based—peak intensity, peak wavelength; shape-based—integrated intensity, their ratio, full-width half maximum, peak normalized intensity at certain wavelengths, and summation of intensity over several spectral bands) and that yield at their output the fiber temperature in the optically probed area on a spider silk fiber. Starting from neural networks trained on fluorescence spectra acquired in steady state temperature conditions, numerical simulations are performed to assess the quality of the reconstruction of dynamical temperature changes that are photothermally induced by illuminating the fiber with periodically intensity-modulated light. Comparison of the five neural networks investigated to multiple types of curve fits showed that using neural networks trained on a combination of the spectral characteristics improves the accuracy over use of a single independent input, with the greatest accuracy observed for inputs that included both intensity-based measurements (peak intensity) and shape-based measurements (normalized intensity at multiple wavelengths), with an ultimate accuracy of 0.29 K via numerical simulation based on experimental observations. The implications are that quantum dots can be used as a more stable and accurate fluorescence thermometer for solid materials and that use of

  20. CdSe/ZnS quantum dot fluorescence spectra shape-based thermometry via neural network reconstruction

    Science.gov (United States)

    Munro, Troy; Liu, Liwang; Glorieux, Christ; Ban, Heng

    2016-06-01

    As a system of interest gets small, due to the influence of the sensor mass and heat leaks through the sensor contacts, thermal characterization by means of contact temperature measurements becomes cumbersome. Non-contact temperature measurement offers a suitable alternative, provided a reliable relationship between the temperature and the detected signal is available. In this work, exploiting the temperature dependence of their fluorescence spectrum, the use of quantum dots as thermomarkers on the surface of a fiber of interest is demonstrated. The performance is assessed of a series of neural networks that use different spectral shape characteristics as inputs (peak-based—peak intensity, peak wavelength; shape-based—integrated intensity, their ratio, full-width half maximum, peak normalized intensity at certain wavelengths, and summation of intensity over several spectral bands) and that yield at their output the fiber temperature in the optically probed area on a spider silk fiber. Starting from neural networks trained on fluorescence spectra acquired in steady state temperature conditions, numerical simulations are performed to assess the quality of the reconstruction of dynamical temperature changes that are photothermally induced by illuminating the fiber with periodically intensity-modulated light. Comparison of the five neural networks investigated to multiple types of curve fits showed that using neural networks trained on a combination of the spectral characteristics improves the accuracy over use of a single independent input, with the greatest accuracy observed for inputs that included both intensity-based measurements (peak intensity) and shape-based measurements (normalized intensity at multiple wavelengths), with an ultimate accuracy of 0.29 K via numerical simulation based on experimental observations. The implications are that quantum dots can be used as a more stable and accurate fluorescence thermometer for solid materials and that use of

  1. Fluorescence applications in molecular neurobiology.

    Science.gov (United States)

    Taraska, Justin W; Zagotta, William N

    2010-04-29

    Macromolecules drive the complex behavior of neurons. For example, channels and transporters control the movements of ions across membranes, SNAREs direct the fusion of vesicles at the synapse, and motors move cargo throughout the cell. Understanding the structure, assembly, and conformational movements of these and other neuronal proteins is essential to understanding the brain. Developments in fluorescence have allowed the architecture and dynamics of proteins to be studied in real time and in a cellular context with great accuracy. In this review, we cover classic and recent methods for studying protein structure, assembly, and dynamics with fluorescence. These methods include fluorescence and luminescence resonance energy transfer, single-molecule bleaching analysis, intensity measurements, colocalization microscopy, electron transfer, and bimolecular complementation analysis. We present the principles of these methods, highlight recent work that uses the methods, and discuss a framework for interpreting results as they apply to molecular neurobiology.

  2. Lasing from fluorescent protein crystals.

    Science.gov (United States)

    Oh, Heon Jeong; Gather, Malte C; Song, Ji-Joon; Yun, Seok Hyun

    2014-12-15

    We investigated fluorescent protein crystals for potential photonic applications, for the first time to our knowledge. Rod-shaped crystals of enhanced green fluorescent protein (EGFP) were synthesized, with diameters of 0.5-2 μm and lengths of 100-200 μm. The crystals exhibit minimal light scattering due to their ordered structure and generate substantially higher fluorescence intensity than EGFP or dye molecules in solutions. The magnitude of concentration quenching in EGFP crystals was measured to be about 7-10 dB. Upon optical pumping at 485 nm, individual EGFP crystals located between dichroic mirrors generated laser emission with a single-mode spectral line at 513 nm. Our results demonstrate the potential of protein crystals as novel optical elements for self-assembled, micro- or nano-lasers and amplifiers in aqueous environment.

  3. Mean fluorescence lifetime and its error

    Energy Technology Data Exchange (ETDEWEB)

    Fiserova, Eva [Department of Mathematical Analysis and Applications of Mathematics, Faculty of Science, Palacky University in Olomouc, tr. 17. listopadu 12, CZE-77146 Olomouc (Czech Republic); Kubala, Martin, E-mail: mkubala@prfnw.upol.cz [Department of Biophysics, Faculty of Science, Palacky University in Olomouc, tr. 17. listopadu 12, CZE-77146 Olomouc (Czech Republic)

    2012-08-15

    Mean excited-state lifetime is one of the fundamental fluorescence characteristics and enters as an important parameter into numerous calculations characterizing molecular interactions, such as e.g. FRET or fluorescence quenching. Our experiments demonstrated that the intensity-weighted mean fluorescence lifetime is very robust characteristic, in contrast to the amplitude-weighted one, which value is dependent on the data quality and particularly on the used fitting model. For the first time, we also report the procedure for the error estimation for both the intensity- and amplitude-weighted mean fluorescence lifetimes. Furthermore, we present a method for estimation of the mean fluorescence lifetime directly from the fluorescence-decay curve recorded by TCSPC (Time-Correlated Single-Photon Counting) method. For its simplicity and low computational demands, it could be a useful tool in the high-throughput applications, such as FACS, FLIM-FRET or HPLC detectors. - Highlights: Black-Right-Pointing-Pointer Intensity-weighted mean fluorescence lifetime is very robust characteristic. Black-Right-Pointing-Pointer The amplitude-weighted mean lifetime depends on the selection of fitting model. Black-Right-Pointing-Pointer Rigorous procedure for estimation of confidence intervals for mean lifetime. Black-Right-Pointing-Pointer The mean lifetime can be estimated directly from the TCSPC histogram.

  4. Fluorescence behaviour and dural infiltration of meningioma analyzed by 5-ALA based fluorescence: Operating microscope versus mini-spectrometer.

    Science.gov (United States)

    Knipps, Johannes; Beseoglu, Kerim; Kamp, Marcel; Fischer, Igor; Felsberg, Joerg; Neumann, Lisa M; Steiger, Hans-Jakob; Cornelius, Jan F

    2017-08-30

    To compare fluorescence intensity of tumor specimens as measured by a FGS-microscope and a spectrometer, 2.) to evaluate tumor infiltration of dura mater around meningiomas with help of these two different 5-ALA based fluorescence tools and 3.) to correlate fluorescence intensity with histopathological data. In a clinical series menigiomas were resected by 5-ALA fluorescence guided surgery. Fluorescence intensity was semi-quantitatively rated by the surgeon at pre-defined points. Biopsies were harvested and fluorescence intensity measured by a spectrometer and histopathologically analysed. Sampling was realized at the level of the dura in a centrifugal direction. 104 biopsies (n= 13 tumors) were analysed. Specificity and sensitivity of the microscope were 0.96 and 0.53 and of the spectrometer 0.95 and 0.93, respectively. Fluorescence intensity as measured by the spectrometer was correlated to histologically confirmed tumor burden. In a centrifugal direction, tumor burden and fluorescence intensity continuously decreased (along the dural tail). Below a threshold value of 639 arbitrary units no tumor was histologically detectable. At the level of the dura the spectrometer was highly sensitive for detection of meningioma cells. The surgical microscope showed false negative results and missed residual tumor cells in more than half of the cases. The complementary use of both fluorescence tools may improve resection quality. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Accurate study of FosPeg® distribution in a mouse model using fluorescence imaging technique and fluorescence white monte carlo simulations

    DEFF Research Database (Denmark)

    Xie, Haiyan; Liu, Haichun; Svenmarker, Pontus

    2010-01-01

    Fluorescence imaging is used for quantitative in vivo assessment of drug concentration. Light attenuation in tissue is compensated for through Monte-Carlo simulations. The intrinsic fluorescence intensity, directly proportional to the drug concentration, could be obtained....

  6. Red and Green Fluorescence from Oral Biofilms

    Science.gov (United States)

    Hoogenkamp, Michel A.; Krom, Bastiaan P.; Janus, Marleen M.; ten Cate, Jacob M.; de Soet, Johannes J.; Crielaard, Wim; van der Veen, Monique H.

    2016-01-01

    Red and green autofluorescence have been observed from dental plaque after excitation by blue light. It has been suggested that this red fluorescence is related to caries and the cariogenic potential of dental plaque. Recently, it was suggested that red fluorescence may be related to gingivitis. Little is known about green fluorescence from biofilms. Therefore, we assessed the dynamics of red and green fluorescence in real-time during biofilm formation. In addition, the fluorescence patterns of biofilm formed from saliva of eight different donors are described under simulated gingivitis and caries conditions. Biofilm formation was analysed for 12 hours under flow conditions in a microfluidic BioFlux flow system with high performance microscopy using a camera to allow live cell imaging. For fluorescence images dedicated excitation and emission filters were used. Both green and red fluorescence were linearly related with the total biomass of the biofilms. All biofilms displayed to some extent green and red fluorescence, with higher red and green fluorescence intensities from biofilms grown in the presence of serum (gingivitis simulation) as compared to the sucrose grown biofilms (cariogenic simulation). Remarkably, cocci with long chain lengths, presumably streptococci, were observed in the biofilms. Green and red fluorescence were not found homogeneously distributed within the biofilms: highly fluorescent spots (both green and red) were visible throughout the biomass. An increase in red fluorescence from the in vitro biofilms appeared to be related to the clinical inflammatory response of the respective saliva donors, which was previously assessed during an in vivo period of performing no-oral hygiene. The BioFlux model proved to be a reliable model to assess biofilm fluorescence. With this model, a prediction can be made whether a patient will be prone to the development of gingivitis or caries. PMID:27997567

  7. Red and Green Fluorescence from Oral Biofilms.

    Science.gov (United States)

    Volgenant, Catherine M C; Hoogenkamp, Michel A; Krom, Bastiaan P; Janus, Marleen M; Ten Cate, Jacob M; de Soet, Johannes J; Crielaard, Wim; van der Veen, Monique H

    2016-01-01

    Red and green autofluorescence have been observed from dental plaque after excitation by blue light. It has been suggested that this red fluorescence is related to caries and the cariogenic potential of dental plaque. Recently, it was suggested that red fluorescence may be related to gingivitis. Little is known about green fluorescence from biofilms. Therefore, we assessed the dynamics of red and green fluorescence in real-time during biofilm formation. In addition, the fluorescence patterns of biofilm formed from saliva of eight different donors are described under simulated gingivitis and caries conditions. Biofilm formation was analysed for 12 hours under flow conditions in a microfluidic BioFlux flow system with high performance microscopy using a camera to allow live cell imaging. For fluorescence images dedicated excitation and emission filters were used. Both green and red fluorescence were linearly related with the total biomass of the biofilms. All biofilms displayed to some extent green and red fluorescence, with higher red and green fluorescence intensities from biofilms grown in the presence of serum (gingivitis simulation) as compared to the sucrose grown biofilms (cariogenic simulation). Remarkably, cocci with long chain lengths, presumably streptococci, were observed in the biofilms. Green and red fluorescence were not found homogeneously distributed within the biofilms: highly fluorescent spots (both green and red) were visible throughout the biomass. An increase in red fluorescence from the in vitro biofilms appeared to be related to the clinical inflammatory response of the respective saliva donors, which was previously assessed during an in vivo period of performing no-oral hygiene. The BioFlux model proved to be a reliable model to assess biofilm fluorescence. With this model, a prediction can be made whether a patient will be prone to the development of gingivitis or caries.

  8. Sound Intensity

    DEFF Research Database (Denmark)

    Crocker, M.J.; Jacobsen, Finn

    1997-01-01

    This chapter is an overview, intended for readers with no special knowledge about this particular topic. The chapter deals with all aspects of sound intensity and its measurement from the fundamental theoretical background to practical applications of the measurement technique....

  9. Plasmonics Enhanced Smartphone Fluorescence Microscopy

    KAUST Repository

    Wei, Qingshan

    2017-05-12

    Smartphone fluorescence microscopy has various applications in point-of-care (POC) testing and diagnostics, ranging from e.g., quantification of immunoassays, detection of microorganisms, to sensing of viruses. An important need in smartphone-based microscopy and sensing techniques is to improve the detection sensitivity to enable quantification of extremely low concentrations of target molecules. Here, we demonstrate a general strategy to enhance the detection sensitivity of a smartphone-based fluorescence microscope by using surface-enhanced fluorescence (SEF) created by a thin metal-film. In this plasmonic design, the samples are placed on a silver-coated glass slide with a thin spacer, and excited by a laser-diode from the backside through a glass hemisphere, generating surface plasmon polaritons. We optimized this mobile SEF system by tuning the metal-film thickness, spacer distance, excitation angle and polarization, and achieved ~10-fold enhancement in fluorescence intensity compared to a bare glass substrate, which enabled us to image single fluorescent particles as small as 50 nm in diameter and single quantum-dots. Furthermore, we quantified the detection limit of this platform by using DNA origami-based brightness standards, demonstrating that ~80 fluorophores per diffraction-limited spot can be readily detected by our mobile microscope, which opens up new opportunities for POC diagnostics and sensing applications in resource-limited-settings.

  10. Sound Intensity

    DEFF Research Database (Denmark)

    Crocker, M.J.; Jacobsen, Finn

    1997-01-01

    This chapter is an overview, intended for readers with no special knowledge about this particular topic. The chapter deals with all aspects of sound intensity and its measurement from the fundamental theoretical background to practical applications of the measurement technique.......This chapter is an overview, intended for readers with no special knowledge about this particular topic. The chapter deals with all aspects of sound intensity and its measurement from the fundamental theoretical background to practical applications of the measurement technique....

  11. Sound intensity

    DEFF Research Database (Denmark)

    Crocker, Malcolm J.; Jacobsen, Finn

    1998-01-01

    This chapter is an overview, intended for readers with no special knowledge about this particular topic. The chapter deals with all aspects of sound intensity and its measurement from the fundamental theoretical background to practical applications of the measurement technique.......This chapter is an overview, intended for readers with no special knowledge about this particular topic. The chapter deals with all aspects of sound intensity and its measurement from the fundamental theoretical background to practical applications of the measurement technique....

  12. A PDMS-Based Cylindrical Hybrid Lens for Enhanced Fluorescence Detection in Microfluidic Systems

    Directory of Open Access Journals (Sweden)

    Bor-Shyh Lin

    2014-02-01

    Full Text Available Microfluidic systems based on fluorescence detection have been developed and applied for many biological and chemical applications. Because of the tiny amount of sample in the system; the induced fluorescence can be weak. Therefore, most microfluidic systems deploy multiple optical components or sophisticated equipment to enhance the efficiency of fluorescence detection. However, these strategies encounter common issues of complex manufacturing processes and high costs. In this study; a miniature, cylindrical and hybrid lens made of polydimethylsiloxane (PDMS to improve the fluorescence detection in microfluidic systems is proposed. The hybrid lens integrates a laser focusing lens and a fluorescence collecting lens to achieve dual functions and simplify optical setup. Moreover, PDMS has advantages of low-cost and straightforward fabrication compared with conventional optical components. The performance of the proposed lens is first examined with two fluorescent dyes and the results show that the lens provides satisfactory enhancement for fluorescence detection of Rhodamine 6G and Nile Red. The overall increments in collected fluorescence signal and detection sensitivity are more than 220% of those without lens, and the detection limits of Rhodamine 6G and Nile red are lowered to 0.01 μg/mL and 0.05 μg/mL, respectively. The hybrid lens is further applied to the detection of Nile red-labeled Chlorella vulgaris cells and it increases both signal intensity and detection sensitivity by more than 520%. The proposed hybrid lens also dramatically reduces the variation in detected signal caused by the deviation in incident angle of excitation light.

  13. A PDMS-based cylindrical hybrid lens for enhanced fluorescence detection in microfluidic systems.

    Science.gov (United States)

    Lin, Bor-Shyh; Yang, Yu-Ching; Ho, Chong-Yi; Yang, Han-Yu; Wang, Hsiang-Yu

    2014-02-13

    Microfluidic systems based on fluorescence detection have been developed and applied for many biological and chemical applications. Because of the tiny amount of sample in the system; the induced fluorescence can be weak. Therefore, most microfluidic systems deploy multiple optical components or sophisticated equipment to enhance the efficiency of fluorescence detection. However, these strategies encounter common issues of complex manufacturing processes and high costs. In this study; a miniature, cylindrical and hybrid lens made of polydimethylsiloxane (PDMS) to improve the fluorescence detection in microfluidic systems is proposed. The hybrid lens integrates a laser focusing lens and a fluorescence collecting lens to achieve dual functions and simplify optical setup. Moreover, PDMS has advantages of low-cost and straightforward fabrication compared with conventional optical components. The performance of the proposed lens is first examined with two fluorescent dyes and the results show that the lens provides satisfactory enhancement for fluorescence detection of Rhodamine 6G and Nile Red. The overall increments in collected fluorescence signal and detection sensitivity are more than 220% of those without lens, and the detection limits of Rhodamine 6G and Nile red are lowered to 0.01 μg/mL and 0.05 μg/mL, respectively. The hybrid lens is further applied to the detection of Nile red-labeled Chlorella vulgaris cells and it increases both signal intensity and detection sensitivity by more than 520%. The proposed hybrid lens also dramatically reduces the variation in detected signal caused by the deviation in incident angle of excitation light.

  14. Study of influencing factors to chromophoric dissolved organic matter absorption properties from fluorescence features in Taihu lake in autumn

    Directory of Open Access Journals (Sweden)

    Chuang-Chun Huang

    2013-04-01

    Full Text Available In order to identify the components of chromophoric dissolved organic matter (CDOM, confirm the influence of components to the absorption coefficient of CDOM (aCDOM, and estimate aCDOM from fluorescence spectra, fluorescence and optical measurements of CDOM were carried out in November 2008. The results indicate that, the primary component of CDOM is humic-like. The secondary component is tryptophan-like, which is the product of phytoplankton and aquatic debris rather than the wastewater treatment drainaged from city. In this study, six fluorophores with multiple excitation-emission matrices (EEMs peaks (A, B, C, N, M, T were identified according to the parallel factor analysis (PARAFAC. The average contribution of each component to the CDOM is 19.93, 18.82, 16.88, 16.39, 12.26, and 15.72%, respectively. Red Shifted phenomenon will happen with the increase of fluorescence intensity for ultraviolet and terrestrially humic-like. Conversely, marine humic-like will appear Reverse Red Shifted with the increase of fluorescence intensity. The primary contributor to the shoulder value of CDOM’s absorption coefficient at 275 nm is phytoplankton productivity, followed by marine humic-like. The main contributors to the shoulder shape are UV humic-like and phytoplankton productivity, followed by marine humic-like and tryptophan-like. A strong correlation between CDOM absorption and fluorescence intensity at emission wavelength of 424 nm and excitation wavelength ranging from 280 to 360 nm was found. The absorption coefficient can be retrieved successfully from the same excitation wavelength’s fluorescence intensity by an exponential model.

  15. Fluorescent molecularly imprinted polymer film binds glucose with a concomitant changes in fluorescence.

    Science.gov (United States)

    Manju, S; Hari, P R; Sreenivasan, K

    2010-10-15

    A fluorescent molecularly imprinted polymeric formulation capable of picking up glucose from aqueous media is reported. The fluorescence intensity of the polymer film was found to reduce proportionally with the concentration of glucose facilitating its use as a glucose sensing element. We used commercially available tear fluid to demonstrate the ability of the film to recognize glucose among other sugar molecules. Fluorescence was measured after equilibrating the film in tear fluid in the presence of a mixture of different sugars. We observed a reduction in fluorescence intensity due to the nonspecific binding of the sugars. The intensity remains the same even if we added additional quantities of the sugars. Interestingly, the fluorescence intensity of the film was found to decrease proportionally when varied concentrations of glucose was added indicating the ability of the film to recognize and bind glucose from a mixture of other sugars. Detectable changes in fluorescence intensity were observed with a concentration of 10 μg/mL of glucose. The results show that the polymer film could be used for detecting glucose in aqueous fluids such as tear. Copyright © 2010 Elsevier B.V. All rights reserved.

  16. Cryo-imaging of fluorescently labeled single cells in a mouse

    Science.gov (United States)

    Steyer, Grant J.; Roy, Debashish; Salvado, Olivier; Stone, Meredith E.; Wilson, David L.

    2009-02-01

    We developed a cryo-imaging system to provide single-cell detection of fluorescently labeled cells in mouse, with particular applicability to stem cells and metastatic cancer. The Case cryoimaging system consists of a fluorescence microscope, robotic imaging positioner, customized cryostat, PC-based control system, and visualization/analysis software. The system alternates between sectioning (10-40 μm) and imaging, collecting color brightfield and fluorescent blockface image volumes >60GB. In mouse experiments, we imaged quantum-dot labeled stem cells, GFP-labeled cancer and stem cells, and cell-size fluorescent microspheres. To remove subsurface fluorescence, we used a simplified model of light-tissue interaction whereby the next image was scaled, blurred, and subtracted from the current image. We estimated scaling and blurring parameters by minimizing entropy of subtracted images. Tissue specific attenuation parameters were found [uT : heart (267 +/- 47.6 μm), liver (218 +/- 27.1 μm), brain (161 +/- 27.4 μm)] to be within the range of estimates in the literature. "Next image" processing removed subsurface fluorescence equally well across multiple tissues (brain, kidney, liver, adipose tissue, etc.), and analysis of 200 microsphere images in the brain gave 97+/-2% reduction of subsurface fluorescence. Fluorescent signals were determined to arise from single cells based upon geometric and integrated intensity measurements. Next image processing greatly improved axial resolution, enabled high quality 3D volume renderings, and improved enumeration of single cells with connected component analysis by up to 24%. Analysis of image volumes identified metastatic cancer sites, found homing of stem cells to injury sites, and showed microsphere distribution correlated with blood flow patterns. We developed and evaluated cryo-imaging to provide single-cell detection of fluorescently labeled cells in mouse. Our cryo-imaging system provides extreme (>60GB), micron

  17. Chemical Address Tags of Fluorescent Bioimaging Probes

    OpenAIRE

    Shedden, Kerby; Rosania, Gus R.

    2010-01-01

    Chemical address tags can be defined as specific structural features shared by a set of bioimaging probes having a predictable influence on cell-associated visual signals obtained from these probes. Here, using a large image dataset acquired with a high content screening instrument, machine vision and cheminformatics analysis have been applied to reveal chemical address tags. With a combinatorial library of fluorescent molecules, fluorescence signal intensity, spectral, and spatial features c...

  18. The Dynamics of Intensive Cultivation

    OpenAIRE

    Christian Bidard

    2008-01-01

    An increase in the demand for agricultural goods leads to the use of more intensive cultivation methods. Though Ricardo sees no difficulties in the intensification process, their existence is revealed by the possible occurrence of multiple equilibria. A general theory of intensive rent is based on a formal parallel with single-product systems without land.

  19. Center for Fluorescence Spectroscopy: advanced studies of fluorescence dynamics, lifetime imaging, clinical sensing, two-photon excitation, and light quenching

    Science.gov (United States)

    Lakowicz, Joseph R.; Malak, Henryk M.; Gryczynski, Ignacy; Szmacinski, Henryk; Kusba, Jozef; Akkaya, Engin; Terpetschnig, Ewald A.; Johnson, Michael L.

    1994-08-01

    The Center for Fluorescence Spectroscopy (CFS) is a multi-user facility providing state of the art time-resolved fluorescence instrumentation and software for scientists, whose research can be enhanced by such experimental data. The CFS is a national center, supported by the National Center for Research Resources Division of the National Institutes of Health, and in part by the National Science Foundation. Both time-domain (TD) and frequency- domain (FD) measurements (10 MHz to 10 Ghz) are available, with a wide range of excitation and emission wavelengths (UV to NIR). The data can be used to recover distances and site-to-site diffusion in protein, interactions between macromolecules, accessibility of fluorophores to quenchers, and the dynamic properties of proteins, membranes and nucleic acids. Current software provides for analysis of multi-exponential intensity and anisotropy decays, lifetime distribution, distance distributions for independent observation of fluorescence donors and acceptors, transient effects in collisional quenching, phase-modulation spectra and time-resolved emission spectra. Most programs provide for global analysis of multiple data sets obtained under similar experimental conditions. Data can be analyzed on-site by connection with the CFS computers through the internet. During six years of operation we have established scientific collaborations with over 30 academic and industrial groups in the United States. These collaborations have resulted in 63 scientific papers.

  20. Fluorescent multiplex cell flow systems and methods

    KAUST Repository

    Merzaban, Jasmeen

    2017-06-01

    Systems and methods are provided for simultaneously assaying cell adhesion or cell rolling for multiple cell specimens. One embodiment provides a system for assaying adhesion or cell rolling of multiple cell specimens that includes a confocal imaging system containing a parallel plate flow chamber, a pump in fluid communication with the parallel plate flow chamber via a flow chamber inlet line and a cell suspension in fluid communication with the parallel plate flow chamber via a flow chamber outlet line. The system also includes a laser scanning system in electronic communication with the confocal imaging system, and a computer in communication with the confocal imaging system and laser scanning system. In certain embodiments, the laser scanning system emits multiple electromagnetic wavelengths simultaneously it cause multiple fluorescent labels having different excitation wavelength maximums to fluoresce. The system can simultaneously capture real-time fluorescence images from at least seven cell specimens in the parallel plate flow chamber.

  1. Fluorescent Probes and Fluorescence (Microscopy Techniques — Illuminating Biological and Biomedical Research

    Directory of Open Access Journals (Sweden)

    Gregor P. C. Drummen

    2012-11-01

    Full Text Available Fluorescence, the absorption and re-emission of photons with longer wavelengths, is one of those amazing phenomena of Nature. Its discovery and utilization had, and still has, a major impact on biological and biomedical research, since it enables researchers not just to visualize normal physiological processes with high temporal and spatial resolution, to detect multiple signals concomitantly, to track single molecules in vivo, to replace radioactive assays when possible, but also to shed light on many pathobiological processes underpinning disease states, which would otherwise not be possible. Compounds that exhibit fluorescence are commonly called fluorochromes or fluorophores and one of these fluorescent molecules in particular has significantly enabled life science research to gain new insights in virtually all its sub-disciplines: Green Fluorescent Protein. Because fluorescent proteins are synthesized in vivo, integration of fluorescent detection methods into the biological system via genetic techniques now became feasible. Currently fluorescent proteins are available that virtually span the whole electromagnetic spectrum. Concomitantly, fluorescence imaging techniques were developed, and often progress in one field fueled innovation in the other. Impressively, the properties of fluorescence were utilized to develop new assays and imaging modalities, ranging from energy transfer to image molecular interactions to imaging beyond the diffraction limit with super-resolution microscopy. Here, an overview is provided of recent developments in both fluorescence imaging and fluorochrome engineering, which together constitute the “fluorescence toolbox” in life science research.

  2. Peptide-stabilized, fluorescent silver nanoclusters

    DEFF Research Database (Denmark)

    Gregersen, Simon; Vosch, Tom André Jos; Jensen, Knud Jørgen

    2016-01-01

    . Herein, we demonstrate how solid-phase methods can increase throughput dramatically in peptide ligand screening and in initial evaluation of fluorescence intensity and chemical stability of peptide-stabilized AgNCs (P-AgNCs). 9-Fluorenylmethyloxycarbonyl (Fmoc) solid-phase peptide synthesis......Few-atom silver nanoclusters (AgNCs) can exhibit strong fluorescence; however, they require ligands to prevent aggregation into larger nanoparticles. Fluorescent AgNCs in biopolymer scaffolds have so far mainly been synthesized in solution, and peptides have only found limited use compared to DNA...

  3. Fluorescent properties of low-molecular-weight fractions from chernozem humic acids

    Science.gov (United States)

    Trubetskoi, O. A.; Demin, D. V.; Trubetskaya, O. E.

    2013-10-01

    The polyacrylamide gel electrophoresis of chernozem humic acids (HAs) followed by ultraviolet detection (λ = 312 nm) has revealed a new highly fluorescent fraction that has the highest electrophoretic mobility and the lowest nominal molecular weight (NMW). The preparative isolation of the fraction has been performed using the multiple microfiltration of the same HA sample in a 7 M carbamide solution on a membrane with a nominal pore size of 5 kDa. Thirty ultrafiltrates with NMW 5 kDa. Fluorescence maximums at and below 490 nm have been noted only in the first four ultrafiltrates. All the ultrafiltrates have been combined into the fraction with NMW membranes of 3 and 1 kDa. Solutions of subfractions F 3-5 kDa, F 1-3 kDa, and F < 1 kDa with fluorescence maximums at 505, 488, and 465 nm, respectively, have been prepared. The F < 1 kDa subfraction with the lowest NMW had the highest fluorescence intensity. The distribution of the fluorescence maximums in the ultrafiltrates has indicated the presence of at least two groups of fluorophores and has confirmed the supramolecular organization of the extracted soil HAs.

  4. Indocyanine green fluorescence angiography for quantitative evaluation of in situ parathyroid gland perfusion and function after total thyroidectomy.

    Science.gov (United States)

    Lang, Brian Hung-Hin; Wong, Carlos K H; Hung, Hing Tsun; Wong, Kai Pun; Mak, Ka Lun; Au, Kin Bun

    2017-01-01

    Because the fluorescent light intensity on an indocyanine green fluorescence angiography reflects the blood perfusion within a focused area, the fluorescent light intensity in the remaining in situ parathyroid glands may predict postoperative hypocalcemia risk after total thyroidectomy. Seventy patients underwent intraoperative indocyanine green fluorescence angiography after total thyroidectomy. Any parathyroid glands with a vascular pedicle was left in situ while any parathyroid glands without pedicle or inadvertently removed was autotransplanted. After total thyroidectomy, an intravenous 2.5 mg indocyanine green fluorescence angiography was given and real-time fluorescent images of the thyroid bed were recorded using the SPY imaging system (Novadaq, Ontario, Canada). The fluorescent light intensity of each indocyanine green fluorescence angiography as well as the average and greatest fluorescent light intensity in each patient were calculated. Postoperative hypocalcemia was defined as adjusted calcium 150% developed postoperative hypocalcemia while 9 (81.8%) patients with a greatest fluorescent light intensity ≤150% did. Similarly, no patients with an average fluorescent light intensity >109% developed PH while 9 (30%) with an average fluorescent light intensity ≤109% did. The greatest fluorescent light intensity was more predictive than day-0 postoperative hypocalcemia (P = .027) and % PTH drop day-0 to 1 (P < .001). Indocyanine green fluorescence angiography is a promising operative adjunct in determining residual parathyroid glands function and predicting postoperative hypocalcemia risk after total thyroidectomy. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Assessing the Transferability of Statistical Predictive Models for Leaf Area Index Between Two Airborne Discrete Return LiDAR Sensor Designs Within Multiple Intensely Managed Loblolly Pine Forest Locations in the South-Eastern USA

    Science.gov (United States)

    Sumnall, Matthew; Peduzzi, Alicia; Fox, Thomas R.; Wynne, Randolph H.; Thomas, Valerie A.; Cook, Bruce

    2016-01-01

    Leaf area is an important forest structural variable which serves as the primary means of mass and energy exchange within vegetated ecosystems. The objective of the current study was to determine if leaf area index (LAI) could be estimated accurately and consistently in five intensively managed pine plantation forests using two multiple-return airborne LiDAR datasets. Field measurements of LAI were made using the LiCOR LAI2000 and LAI2200 instruments within 116 plots were established of varying size and within a variety of stand conditions (i.e. stand age, nutrient regime and stem density) in North Carolina and Virginia in 2008 and 2013. A number of common LiDAR return height and intensity distribution metrics were calculated (e.g. average return height), in addition to ten indices, with two additional variants, utilized in the surrounding literature which have been used to estimate LAI and fractional cover, were calculated from return heights and intensity, for each plot extent. Each of the indices was assessed for correlation with each other, and was used as independent variables in linear regression analysis with field LAI as the dependent variable. All LiDAR derived metrics were also entered into a forward stepwise linear regression. The results from each of the indices varied from an R2 of 0.33 (S.E. 0.87) to 0.89 (S.E. 0.36). Those indices calculated using ratios of all returns produced the strongest correlations, such as the Above and Below Ratio Index (ABRI) and Laser Penetration Index 1 (LPI1). The regression model produced from a combination of three metrics did not improve correlations greatly (R2 0.90; S.E. 0.35). The results indicate that LAI can be predicted over a range of intensively managed pine plantation forest environments accurately when using different LiDAR sensor designs. Those indices which incorporated counts of specific return numbers (e.g. first returns) or return intensity correlated poorly with field measurements. There were

  6. The use of fluorescence correlation spectroscopy to probe mitochondrial mobility and intramatrix protein diffusion

    NARCIS (Netherlands)

    P.H.G.M. Willems; H.G. Swarts; M.A. Hink; W.J.H. Koopman

    2009-01-01

    Within cells, functional changes in mitochondrial metabolic state are associated with alterations in organelle mobility, shape, and configuration of the mitochondrial matrix. Fluorescence correlation spectroscopy (FCS) is a technique that measures intensity fluctuations caused by single fluorescent

  7. Fundamentals of fluorescence and fluorescence microscopy.

    Science.gov (United States)

    Wolf, David E

    2013-01-01

    This chapter discusses the fundamental physics of fluorescence. The application of fluorescence to microscopy represents an important transition in the development of microscopy, particularly as it applies to biology. It enables quantitating the amounts of specific molecules within a cell, determining whether molecules are complexing on a molecular level, measuring changes in ionic concentrations within cells and organelles, and measuring molecular dynamics. This chapter also discusses the issues important to quantitative measurement of fluorescence and focuses on four of quantitative measurements of fluorescence--boxcar-gated detection, streak cameras, photon correlation, and phase modulation. Although quantitative measurement presents many pitfalls to the beginner, it also presents significant opportunities to one skilled in the art. This chapter also examines how fluorescence is measured in the steady state and time domain and how fluorescence is applied in the modern epifluorescence microscope.

  8. Alloying effect on K shell X-ray fluorescence cross-sections and intensity ratios of Cu and Sn in Cu1Sn1-x alloys using the 59.5 keV gamma rays

    Science.gov (United States)

    Dogan, M.; Olgar, M. A.; Cengiz, E.; Tıraşoglu, E.

    2016-09-01

    Kβ/Kα, intensity ratios and σKα,β production cross-sections of Cu and Sn were measured in pure metals and in different alloys which have different compositions (CuxSn1-x x=0.48, 0.41, 0.14 and 0.06). The samples were excited by 59.5 keV γ-rays from 241Am annular radioactive source. K X-rays emitted by samples were counted by an Ultra-LEGe detector with a resolution of 150 eV at 5.9 keV. Comparison of the σKβ production cross-sections and Kβ/Kα X-ray intensity ratio values for Cu and Sn with the theoretical and semi-empirical calculations indicates that they are in the inverse direction with concentration of constituent element in the alloys. The results show that variations in these parameters can be explained with the charge transfer process between the elements which constitute the alloys.

  9. Terapia fotodinâmica com ftalocianina de zinco tópica: avaliação da intensidade de fluorescência, absorção cutânea, alterações histológicas e imuno-histoquímicas na pele do modelo animal Topical photodynamic therapy with zinc phthalocyanine: evaluation of fluorescence intensity, skin absorption, skin histological and immunohistochemical changes in animal model

    Directory of Open Access Journals (Sweden)

    Marília Vannuchi Tomazini

    2007-12-01

    . After an 8-hour occlusion of different formulations, the mice were exposed to 670 nm laser, at a 50 J.cm-² dose. RESULTS - Fluorescence was slightly higher after 8 hours, and also with emulsion formulation at one-, two- and four-hour occlusion periods. The intensity of edema and erosion were correlated to epidermal necrosis and to Fas immunoexpression in skin histological specimens. CONCLUSIONS - The results show the effects of photodynamic action promoted by the interaction between Zn-PC formulation and a 670-nm light source. Macro and micromorphological alterations were correlated and more substantial with monolein and Zn-PC emulsion, suggesting more marked effects with this formulation. The Fas immunoexpression and histological changes suggested that apoptosis plays a role in the mechanism of cell death caused by PDT based in Zn-PC.

  10. Fluorescence emission of pyrene in surfactant solutions.

    Science.gov (United States)

    Piñeiro, Lucas; Novo, Mercedes; Al-Soufi, Wajih

    2015-01-01

    The systematic description of the complex photophysical behaviour of pyrene in surfactant solutions in combination with a quantitative model for the surfactant concentrations reproduces with high accuracy the steady-state and the time resolved fluorescence intensity of pyrene in surfactant solutions near the cmc, both in the monomer and in the excimer emission bands. We present concise model equations that can be used for the analysis of the pyrene fluorescence intensity in order to estimate fundamental parameters of the pyrene-surfactant system, such as the binding equilibrium constant K of pyrene to a given surfactant micelle, the rate constant of excimer formation in micelles, and the equilibrium constant of pyrene-surfactant quenching. The values of the binding equilibrium constant K(TX100)=3300·10³ M⁻¹ and K(SDS)=190·10³ M⁻¹ for Triton X-100 (TX100) and SDS micelles, respectively, show that the partition of pyrene between bulk water and micelles cannot be ignored, even at relatively high surfactant concentrations above the cmc. We apply the model to the determination of the cmc from the pyrene fluorescence intensity, especially from the intensity ratio at two vibronic bands in the monomer emission or from the ratio of excimer to monomer emission intensity. We relate the finite width of the transition region below and above the cmc with the observed changes in the pyrene fluorescence in this region.

  11. Fabrication of Indocyanine Green and 2H, 3H-perfluoropentane loaded microbubbles for fluorescence and ultrasound imaging

    Science.gov (United States)

    He, Yutong; Wu, Qiang; Ma, Rong; Chang, Shufang; Shao, Pengfei; Xu, Ronald

    2016-03-01

    As a near-infrared (NIR) fluorescence dye, Indocyanine Green (ICG) has not gained broader clinical applications, owing to its multiple limitations such as concentration-dependent aggregation, low fluorescence quantum yield, poor physicochemical stability and rapid elimination from the body. In the meanwhile, 2H,3H-perfluoropentane (H-PFP) has been widely studied in ultrasound imaging as a vehicle for targeted delivery of contrast agents and drugs. We synthesized a novel dual-modal fluorescence and ultrasound contrast agent by encapsulating ICG and H-PFP in lipid microbubbles using a liquid-driven coaxial flow focusing (LDCFF) process. Uniform microbubbles with the sizes ranging from 1-10um and great ICG loading efficiency was achieved by this method. Our benchtop experiments showed that ICG/H-PFP microbubbles exhibited less aggregation, increased fluorescence intensity and more stable photostability compared to free ICG aqueous solution. Our phantom experiments demonstrated that ICG/H-PFP microbubbles enhanced the imaging contrasts in fluorescence imaging and ultrasonography. Our animal experiments indicated that ICG/H-PFP microbubbles extended the ICG life time and facilitated dual mode fluorescence and ultrasound imaging in vivo.

  12. Fluorescent Labeling of Nanometer Hydroxyapatite

    Institute of Scientific and Technical Information of China (English)

    Yuan ZHANG; Yuan YUAN; Changsheng LIU

    2008-01-01

    A novel surface treatment method using 3-aminopropyltriethoxysilane (AMPTES), was developed to immobilize the fluorescein molecule on nano-HAP (nanometer hydroxyapatite) powders. By pretreating the nano-HAP powders surface with AMPTES, fluorescein, chosen on the basis of the chemical structure of the nano- HAP powders, could be bound to the nano-HAP powders surface. The chemical compositions of nano-HAP before and after being labeled were characterized by Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS). The morphology, phase composition, and the fluorescence characteristics of the nano-HAP powders with and without staining were also investigated. The FTIR and XPS results revealed that fiuorescein had been successfully immobilized on the surface of AMPTES-bound nano-HAP powders via the acylamide bond formation between the -COOH of fluorescein and the -NH2 of AMPTES. The labeled nano-HAP powders possessed strong fluorescent intensity with a little deviation from the maximum emission wavelength of fluorescein. But the morphology and phase composition had no obvious alteration. Under fluorescence microscopy, the labeled nano-HAP powders., even after 24 h cell incubation, exhibited strong fluorescence.

  13. Effects of growth characteristics and chlorophyll fluorescence characteristics in leaves of Ocimum basilicum L. seedlings under low light intensity%弱光环境对罗勒幼苗叶片生长和叶绿素荧光特性的影响

    Institute of Scientific and Technical Information of China (English)

    林晗婧; 田野; 张秀丽; 孙广玉

    2015-01-01

    以花色素苷含量不同的紫罗勒和大叶罗勒为对象,分别对其进行弱光和自然光处理30d后进行生长参数、色素含量和叶绿素荧光参数的测定,结果显示:弱光导致两种罗勒幼苗叶片比叶面积、叶面积、叶宽、叶长和株高显著增加,幼苗根长,地上、地下生物量和总生物量明显降低。经过弱光处理的叶片花色苷与总叶绿素比和花色苷含量均降低。总叶绿素含量、叶绿素a含量、叶绿素b含量和叶绿素a/b比值较自然光明显升高。在弱光下,两种罗勒幼苗叶片失活PSⅡ反应中心的热耗散量子产额(ФNF)降低,而基本的荧光量子产额和热耗散的量子产额(Фf,D)升高。说明,花色素苷可能在光破坏防御上发挥了作用。%Based on Ocimum basilicum L. seedlings with different content of anthocyanins, this article studied on the growth parameters, pigment content and chlorophyll fluorescence parameters dealed with low light intensity and nature light intensity for 30 days. Finally, the specific leaf area, leaf area, blade width, leaf length, plant height of leaves of Ocimum basilicum L. seedlings increased significantly, however root length, aboveground biomass, underground biomass, total biomass of leaves decreased significantly with low light intensity. The content of anthocyanins and the ratio of anthocyanins and total chlorophyll of Ocimum basilicum L. seedlings with low light intensity was lower than the one with nature light intensity,however the content of total chlorophyll, chlorophyll a, chlorophyll b, the ratio of chlorophyll a and b of Ocimum basilicum L. seedlings with low light intensity was higher. Dealed with low light intensity, the quantum yield of thermal dissipation in nonfunctional PSⅡ (ФNF) decreased, the basic quantum yield of fluorescence and thermal dissipation(Фf,D) increased in leaves of Ocimum basilicum L. seedlings. The result showed that anthocyanins may have the effect

  14. Reviews in fluorescence 2010

    CERN Document Server

    Geddes, Chris D

    2011-01-01

    ""Reviews in Fluorescence 2010"", the seventh volume of the book serial from Springer, serves as a comprehensive collection of current trends and emerging hot topics in the field of fluorescence and closely related disciplines. It summarizes the year's progress in fluorescence and its applications, with authoritative analytical reviews specialized enough to be attractive to professional researchers, yet also appealing to the wider audience of scientists in related disciplines of fluorescence. ""Reviews in Fluorescence"" offers an essential reference material for any lab working in the fluoresc

  15. Principles of fluorescence techniques

    CERN Document Server

    2016-01-01

    Fluorescence techniques are being used and applied increasingly in academics and industry. The Principles of Fluorescence Techniques course will outline the basic concepts of fluorescence techniques and the successful utilization of the currently available commercial instrumentation. The course is designed for students who utilize fluorescence techniques and instrumentation and for researchers and industrial scientists who wish to deepen their knowledge of fluorescence applications. Key scientists in the field will deliver theoretical lectures. The lectures will be complemented by the direct utilization of steady-state and lifetime fluorescence instrumentation and confocal microscopy for FLIM and FRET applications provided by leading companies.

  16. Technical principles and neurosurgical applications of fluorescein fluorescence using a microscope-integrated fluorescence module.

    Science.gov (United States)

    Rey-Dios, Roberto; Cohen-Gadol, Aaron A

    2013-04-01

    Fluorescent technology has recently become a valuable tool in the surgical management of neoplastic and vascular lesions. The availability of microscope-integrated fluorescent modules has facilitated incorporation of this technology within the microsurgical workflow. The currently available microscope integrated modules use 5-aminolevulinic acid (5-ALA) and indocyanine green (ICG) as fluorophores. Fluorescein sodium is a fluorescent molecule that has been used specifically in ophthalmology for the treatment of retinal angiography. A new microscope-integrated fluorescent module has been recently developed for fluorescein. We employed this technology to maximize resection of tumors and perform intraoperative angiography to guide microsurgical management of aneurysms and arteriovenous malformations. Fluorescein fluorescence allows the surgeon to appreciate fluorescent structures through the oculars while visualizing non-fluorescent tissues in near natural colors. Therefore, the operator can proceed with microsurgery under the fluorescent mode. We present three representative cases in which the use of fluorescein fluorescence was found useful in the surgeon's decision making during surgery. The applications of this new microscope-integrated fluorescent module are multiple, and include vascular and oncologic neurosurgery. Further clinical investigations with large patient cohorts are needed to fully establish the role of this new technology.

  17. Fluorescence enhancement of CdTe/CdS quantum dots by coupling of glyphosate and its application for sensitive detection of copper ion

    Energy Technology Data Exchange (ETDEWEB)

    Liu Zhengqing; Liu Shaopu; Yin Pengfei [Key Laboratory on Luminescence and Real-Time Analysis, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); He Youqiu, E-mail: heyq@swu.edu.cn [Key Laboratory on Luminescence and Real-Time Analysis, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China)

    2012-10-01

    Graphical abstract: Glyphosate (Glyp) had been used to modify the surface of CdTe/CdS QDs, resulting in the enhancement of fluorescence intensity. The Glyp-functionalized QDs fluorescent probe offers good sensitivity and selectivity for detecting Cu{sup 2+} based on the fluorescence quenching. Highlights: Black-Right-Pointing-Pointer Water soluble CdTe/CdS quantum dots capped with glyphosate were firstly synthesized. Black-Right-Pointing-Pointer The fluorescence of the Glyp-functionalized QDs was quenched by copper ion. Black-Right-Pointing-Pointer A new fluorescent sensor for copper ion was developed based on the prepared QDs. Black-Right-Pointing-Pointer The sensor exhibited high sensitivity and good selectivity for copper ion. - Abstract: A novel fluorescent probe for Cu{sup 2+} determination based on the fluorescence quenching of glyphosate (Glyp)-functionalized quantum dots (QDs) was firstly reported. Glyp had been used to modify the surface of QDs to form Glyp-functionalized QDs following the capping of thioglycolic acid on the core-shell CdTe/CdS QDs. Under the optimal conditions, the response was linearly proportional to the concentration of Cu{sup 2+} between 2.4 Multiplication-Sign 10{sup -2} {mu}g mL{sup -1} and 28 {mu}g mL{sup -1}, with a detection limit of 1.3 Multiplication-Sign 10{sup -3} {mu}g mL{sup -1} (3{delta}). The Glyp-functionalized QDs fluorescent probe offers good sensitivity and selectivity for detecting Cu{sup 2+}. The fluorescent probe was successfully used for the determination of Cu{sup 2+} in environmental samples. The mechanism of reaction was also discussed.

  18. Laser induced fluorescence photobleaching anemometer for microfluidic devices.

    Science.gov (United States)

    Wang, G R

    2005-04-01

    We have developed a novel, non-intrusive fluid velocity measurement method based on photobleaching of a fluorescent dye for microfluidic devices. The residence time of the fluorescent dye in a laser beam depends on the flow velocity and approximately corresponds to the decaying time of the photobleaching of the dye in the laser beam. The residence time is inversely proportional to the flow velocity. The fluorescence intensity increases with the flow velocity due to the decrease of the residence time. A calibration curve between fluorescence intensity and known flow velocity should be obtained first. The calibration relationship is then used to calculate the flow velocity directly from the measured fluorescence intensity signal. The new method can measure the velocity very quickly and is easy to use. It is demonstrated for both pressure driven flow and electroosmotic flow.

  19. Fluorescence Spectra Studies on the Interaction between Lanthanides and Calmodulin

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    The conformation of Calmodulin(CaM) induced by lanthanides has been examined using fluorescence methods.With the addition of lanthanide (Ln3+), the intrinsic fluorescence intensity of CaM without calcium ions (Apo-CaM) first increases and then decreases.Ln3+ causes the decrease of intrinsic fluorescence intensity of calcium saturated CaM (Ca2+4-CaM) only at high concentrations.At low concentrations, Ln3+ results not only in the enhancement of fluorescence intensity of Apo-CaM, but also in a blue shift of the maximum emission wavelengh of dansyl labeled calmodulin(Apo-D-CaM).The molecular mechanism of the interaction between Ln3+ and CaM has been discussed in the light of the fluorescence spectra.

  20. Fluorescence spectroscopy for neoplasms control

    Science.gov (United States)

    Bratchenko, I. A.; Kristoforova, Yu. A.; Myakinin, O. O.; Artemyev, D. N.; Kozlov, S. V.; Moryatov, A. A.; Zakharov, V. P.

    2016-04-01

    Investigation of malignant skin tumors diagnosis was performed involving two setups for native tissues fluorescence control in visible and near infrared regions. Combined fluorescence analysis for skin malignant melanomas and basal cell carcinomas was performed. Autofluorescence spectra of normal skin and oncological pathologies stimulated by 457 nm and 785 nm lasers were registered for 74 skin tissue samples. Spectra of 10 melanomas and 27 basal cell carcinomas were registered ex vivo. Skin tumors analysis was made on the basis of autofluorescence spectra intensity and curvature for analysis of porphyrins, lipo-pigments, flavins and melanin. Separation of melanomas and basal cell carcinomas was performed on the basis of discriminant analysis. Overall accuracy of basal cell carcinomas and malignant melanomas separation in current study reached 86.5% with 70% sensitivity and 92.6% specificity.

  1. IR-stimulated visible fluorescence in pink and brown diamond.

    Science.gov (United States)

    Byrne, K S; Chapman, J G; Luiten, A N

    2014-03-19

    Irradiation of natural pink and brown diamond by middle-ultraviolet light (photon energy ϵ ≥ 4.1 eV ) is seen to induce anomalous fluorescence phenomena at N3 defect centres (structure N3-V). When diamonds primed in this fashion are subsequently exposed to infrared light (even with a delay of many hours), a transient burst of blue N3 fluorescence is observed. The dependence of this IR-triggered fluorescence on pump wavelength and intensity suggest that this fluorescence phenomena is intrinsically related to pink diamond photochromism. An energy transfer process between N3 defects and other defect species can account for both the UV-induced fluorescence intensity changes, and the apparent optical upconversion of IR light. From this standpoint, we consider the implications of this N3 fluorescence behaviour for the current understanding of pink diamond photochromism kinetics.

  2. Fluorescence imaging of soybean flavonol isolines

    Science.gov (United States)

    Kim, Moon S.; Lee, Edward H.; Mulchi, Charles L.; McMurtrey, James E., III; Chappelle, Emmett W.; Rowland, Randy A.

    1998-07-01

    Experiments were conducted to characterize the fluorescence emission of leaves from four soybean ('Harosoy') plants containing different concentrations of flavonols (kaempferol glycosides). The investigation utilized genetically mutated soybean flavonol isolines grown in a constant environment, thus limiting factors known to affect fluorescence emission characteristics other than different kaempferol glycosides concentrations. Flavonol isolines included OX922, OX941, OX942, OX944. The first two isolines contain kaempferol (K) glycosides; K3, K6, and K9, and the latter two did not have K3, K6, and K9. A fluorescence imaging system (FIS) was used to characterize steady state florescence images of the sample leaves measured at wavelengths centered at 450, 550, 680, and 740 nm with an excitation at 360 nm. Images taken with FIS greatly complement non-imaging fluorescence measurements by characterizing the spatial variation of fluorescence within leaves. We also acquired fluorescence emission spectra to characterize spectral features of the soybean flavonol isolines. The emission spectral shape of the fluorescence emission characteristics were not significantly different between the soybeans that contain kaempferol glycosides and the ones that do not contain kaempferol glycosides. Typical emission maxima of green vegetation in the blue, green, red, and far-red bands were noticed in all four soybean isolines. However, plants containing kaempferol glycosides, OX922 and OX941 had significantly lower intensities throughout the wavelength regions. These results imply that fluorescence emission intensities in the fluorescence emission bands studied are significantly affected by the presence and absence of kaempferol glycosides concentrations (UV radiation screening compounds). Pure kaempferol glycoside dissolved in solution show minimal fluorescence emission when excited with the absorption maximum radiation at 365 nm. However, a broad band emission can be seen in the green

  3. Fluorescence correlation spectroscopy

    NARCIS (Netherlands)

    Hink, M.A.; Verveer, P.J.

    2015-01-01

    Fluorescence fluctuation spectroscopy techniques allow the quantification of fluorescent molecules present at the nanomolar concentration level. After a brief introduction to the technique, this chapter presents a protocol including background information in order to measure and quantify the molecul

  4. Chemical approaches for mimicking logic functions within fluorescent MPT dyes

    Institute of Scientific and Technical Information of China (English)

    2009-01-01

    The progress of the design, synthesis, fluorescence properties and application of a new family of fluorescent molecular switches towards information processing at the molecular level was reviewed. On the basis of the high fluorescence quantum yields and surroundings-sensitive fluorescent properties of the 5-methoxy-2-(2-pyridyl)-thiazole (2-MPT, 1) and a series of its derivatives as prepared, multiple binary logic and arithmetic functionalities were realized through encoding the controllable fluorescence switching properties with binary digit. Combined with the microfluidic platform, the fabrication of the molecular logic devices was attempted.

  5. Chemical approaches for mimicking logic functions within fluorescent MPT dyes

    Institute of Scientific and Technical Information of China (English)

    XU ChunHu; SUN Wei; ZHANG Chao; BAI YanChun; FANG ChenJie; LI WenTao; HUANG YanYi; YAN ChunHua

    2009-01-01

    The progress of the design, synthesis, fluorescence properties and application of a new family of fluorescent molecular switches towards information processing at the molecular level was reviewed. On the basis of the high fluorescence quantum yields and surroundings-sensitive fluorescent proper-ties of the 5-methoxy-2-(2-pyridyl)-thiazole (2-MPT, 1) and a series of its derivatives as prepared, multi-ple binary logic and arithmetic functionalities were realized through encoding the controllable fluo-rescence switching properties with binary digit. Combined with the microfluidic platform, the fabrica-tion of the molecular logic devices was attempted.

  6. Safe biodegradable fluorescent particles

    Science.gov (United States)

    Martin, Sue I.; Fergenson, David P.; Srivastava, Abneesh; Bogan, Michael J.; Riot, Vincent J.; Frank, Matthias

    2010-08-24

    A human-safe fluorescence particle that can be used for fluorescence detection instruments or act as a safe simulant for mimicking the fluorescence properties of microorganisms. The particle comprises a non-biological carrier and natural fluorophores encapsulated in the non-biological carrier. By doping biodegradable-polymer drug delivery microspheres with natural or synthetic fluorophores, the desired fluorescence can be attained or biological organisms can be simulated without the associated risks and logistical difficulties of live microorganisms.

  7. Intensive mobilities:

    DEFF Research Database (Denmark)

    Vannini, Phillip; Bissell, David; Jensen, Ole B.

    This paper explores the intensities of long distance commuting journeys as a way of exploring how bodily sensibilities are being changed by the mobilities that they undertake. The context of this paper is that many people are travelling further to work than ever before owing to a variety of facto....... By exploring how experiences of long-distance workers become constituted by a range of different material forces enables us to more sensitively consider the practical, technical, and political implications of this increasingly prevalent yet underexplored regime of work....... which relate to transport, housing and employment. Yet we argue that the experiential dimensions of long distance mobilities have not received the attention that they deserve within geographical research on mobilities. This paper combines ideas from mobilities research and contemporary social theory...... with fieldwork conducted in Canada, Denmark and Australia to develop our understanding of the experiential politics of long distance workers. Rather than focusing on the extensive dimensions of mobilities that are implicated in patterns and trends, our paper turns to the intensive dimensions of this experience...

  8. Optical antenna design for fluorescence enhancement in the ultraviolet.

    Science.gov (United States)

    Jiao, Xiaojin; Blair, Steve

    2012-12-31

    Through rational design, we compare the performance of three plasmonic antenna structures for UV fluorescence enhancement. Among the antenna performance metrics considered are the local increase in excitation intensity and the increase in quantum efficiency, the product of which represents the net fluorescence enhancement. With realistic structures in aluminum, we predict that greater than 100× net enhancement can be obtained.

  9. Detection of rheumatoid arthritis in humans by fluorescence imaging

    Science.gov (United States)

    Ebert, Bernd; Dziekan, Thomas; Weissbach, Carmen; Mahler, Marianne; Schirner, Michael; Berliner, Birgitt; Bauer, Daniel; Voigt, Jan; Berliner, Michael; Bahner, Malte L.; Macdonald, Rainer

    2010-02-01

    The blood pool agent indo-cyanine green (ICG) has been investigated in a prospective clinical study for detection of rheumatoid arthritis using fluorescence imaging. Temporal behavior as well as spatial distribution of fluorescence intensity are suited to differentiate healthy and inflamed finger joints after i.v. injection of an ICG bolus.

  10. Enhanced fluorescence of silver nanoclusters stabilized with branched oligonucleotides.

    Science.gov (United States)

    Latorre, Alfonso; Lorca, Romina; Zamora, Félix; Somoza, Álvaro

    2013-05-28

    DNA stabilized silver nanoclusters (AgNCs) are promising optical materials, whose fluorescence properties can be tuned by the selection of the DNA sequence employed. In this work we have used modified oligonucleotides in the preparation of AgNCs. The fluorescent intensity obtained was 60 times higher than that achieved with standard oligonucleotides.

  11. Fluorescence Hybridization Assay Based On Chitosan-Linked Softarrays

    Science.gov (United States)

    2003-07-01

    was incubated in the wells to reduce the Schiff base resulting from the reaction of aldehyde and amine groups. After this reaction, the yellowish...color representative of a Schiff base disappeared and the background fluorescence signal dropped to the initial ~8 to 12 fluorescence intensity (FI

  12. Red fluorescent biofilm: the thick, the old, and the cariogenic

    Directory of Open Access Journals (Sweden)

    Catherine M.C. Volgenant

    2016-04-01

    Full Text Available Background: Some dental plaque fluoresces red. The factors involved in this fluorescence are yet unknown. Objective: The aim of this study was to assess systematically the effect of age, thickness, and cariogenicity on the extent of red fluorescence produced by in vitro microcosm biofilms. Design: The effects of biofilm age and thickness on red fluorescence were tested in a constant depth film fermentor (CDFF by growing biofilms of variable thicknesses that received a constant supply of defined mucin medium (DMM and eight pulses of sucrose/day. The influence of cariogenicity on red fluorescence was tested by growing biofilm on dentin disks receiving DMM, supplemented with three or eight pulses of sucrose/day. The biofilms were analyzed at different time points after inoculation, up to 24 days. Emission spectra were measured using a fluorescence spectrophotometer (λexc405 nm and the biofilms were photographed with a fluorescence camera. The composition of the biofilms was assessed using 454-pyrosequecing of the 16S rDNA gene. Results: From day 7 onward, the biofilms emitted increasing intensities of red fluorescence as evidenced by the combined red fluorescence peaks. The red fluorescence intensity correlated with biofilm thickness but not in a linear way. Biofilm fluorescence also correlated with the imposed cariogenicity, evidenced by the induced dentin mineral loss. Increasing the biofilm age or increasing the sucrose pulsing frequency led to a shift in the microbial composition. These shifts in composition were accompanied by an increase in red fluorescence. Conclusions: The current study shows that a thicker, older, or more cariogenic biofilm results in a higher intensity of red fluorescence.

  13. Red fluorescent biofilm: the thick, the old, and the cariogenic

    Science.gov (United States)

    Volgenant, Catherine M.C.; Hoogenkamp, Michel A.; Buijs, Mark J.; Zaura, Egija; ten Cate, Jacob (Bob) M.; van der Veen, Monique H.

    2016-01-01

    Background Some dental plaque fluoresces red. The factors involved in this fluorescence are yet unknown. Objective The aim of this study was to assess systematically the effect of age, thickness, and cariogenicity on the extent of red fluorescence produced by in vitro microcosm biofilms. Design The effects of biofilm age and thickness on red fluorescence were tested in a constant depth film fermentor (CDFF) by growing biofilms of variable thicknesses that received a constant supply of defined mucin medium (DMM) and eight pulses of sucrose/day. The influence of cariogenicity on red fluorescence was tested by growing biofilm on dentin disks receiving DMM, supplemented with three or eight pulses of sucrose/day. The biofilms were analyzed at different time points after inoculation, up to 24 days. Emission spectra were measured using a fluorescence spectrophotometer (λexc405 nm) and the biofilms were photographed with a fluorescence camera. The composition of the biofilms was assessed using 454-pyrosequecing of the 16S rDNA gene. Results From day 7 onward, the biofilms emitted increasing intensities of red fluorescence as evidenced by the combined red fluorescence peaks. The red fluorescence intensity correlated with biofilm thickness but not in a linear way. Biofilm fluorescence also correlated with the imposed cariogenicity, evidenced by the induced dentin mineral loss. Increasing the biofilm age or increasing the sucrose pulsing frequency led to a shift in the microbial composition. These shifts in composition were accompanied by an increase in red fluorescence. Conclusions The current study shows that a thicker, older, or more cariogenic biofilm results in a higher intensity of red fluorescence. PMID:27060056

  14. Intensive Farmaco- Surveillance of Interferon Alpha 2b Recombinant in Multiple Sclerosis. Farmacovigilancia intensiva al interferón alfa 2b recombinante en la esclerosis múltiple.

    Directory of Open Access Journals (Sweden)

    Hailen Bobillo López

    Full Text Available

    Background: The interferon alpha 2b recombinant, produced in Cuba, is used in the treatment of different illnesses, such as the multiple sclerosis. For its commercialization it is needed to know its safety scope. Objective: To assess the adverse reactions of interferon alpha 2b recombinant in the treatment of multiple sclerosis. Method: During the period between 1996 and 2006, 70 clinical histories and data collection notebooks of patients included in the randomized double blind national clinical trial phase IV were revised. This trial was developed in the Clinic of Multiple Sclerosis of the Hospital "Dr. Gustavo Aldereguía Lima" in Cienfuegos. With regard to the total of manifested adverse reactions we analyzed type, length, and use of a given treatment to counteract them, intensity level (light, moderate, serious or lethal and causality level (definitive, probable, possible, conditional or not related. Results: 53 patients out of the total presented 207 adverse reactions to interferon. The most frequent were: fever, migraine, chills, arthralgia, asthenia and myalgia, being most of them moderate collateral effects of definitive character. In 197 patients the outcome was favorable. Conclusion: The use of the interferon alpha 2b

  15. Conjugation of fluorescent proteins with DNA oligonucleotides.

    Science.gov (United States)

    Lapiene, Vidmantas; Kukolka, Florian; Kiko, Kathrin; Arndt, Andreas; Niemeyer, Christof M

    2010-05-19

    This work describes the synthesis of covalent ssDNA conjugates of six fluorescent proteins, ECFP, EGFP, E(2)GFP, mDsRed, Dronpa, and mCherry, which were cloned with an accessible C-terminal cystein residue to enable site-selective coupling using a heterobispecific cross-linker. The resulting conjugates revealed similar fluorescence emission intensity to the unconjugated proteins, and the functionality of the tethered oligonucleotide was proven by specific Watson-Crick base pairing to cDNA-modified gold nanoparticles. Fluorescence spectroscopy analysis indicated that the fluorescence of the FP is quenched by the gold particle, and the extent of quenching varied with the intrinsic spectroscopic properties of FP as well as with the configuration of surface attachment. Since this study demonstrates that biological fluorophores can be selectively incorporated into and optically coupled with nanoparticle-based devices, applications in DNA-based nanofabrication can be foreseen.

  16. Quantitative X-ray fluorescence analysis of samples of less than `infinite thickness': Difficulties and possibilities

    Science.gov (United States)

    Sitko, Rafał

    2009-11-01

    X-ray fluorescence spectrometry due to its nondestructive nature is widely applied in analysis of single layers and multiple layer films (e.g. semiconductors, electrooptic and solar cell devices, coatings, corrosion and paint layers), individual particles (airborne, fly ash, gunshot residue particles, etc.), art and archeological objects (manuscripts, paintings, icons) and many others. Quantitative analysis of these materials, frequently classified as samples of less than infinite thickness (thin or intermediate-thickness samples), required applying adequate matrix correction methods taking into account complex dependence of analyte fluorescent radiation intensity on full matrix composition and sample thickness. In this article, the matrix correction methods including fundamental parameters, Monte Carlo simulations, influence coefficients algorithms and methods based on X-ray transmission measurements are reviewed. The difficulties in the analysis of single layer and multiple layer films and the accuracy of fundamental parameter methods in simultaneous determination of their thickness and composition are discussed. The quantitative analysis of individual particles and inhomogeneous and/or complex structure materials using fundamental parameter and Monte Carlo simulation methods in micro-beam X-ray fluorescence spectrometry are also reviewed. Some references are devoted to the analysis of light matrix samples, e.g. geological, environmental and biological samples, in which undetectable low-Z elements are present (so-called 'dark matrix') using backscattered fundamental parameter methods. Since the samples of less than infinite thickness are partially transparent for X-ray beams, the transmission measurements present possibilities that are unattainable for bulk samples. Thus, the emission-transmission method and also new instruments allowing measurements of the primary X-ray beam transmitted through the sample together with measurements of X-ray fluorescence

  17. One- and two-photon excited fluorescence lifetimes and anisotropy decays of green fluorescent proteins.

    OpenAIRE

    2000-01-01

    We have used one- (OPE) and two-photon (TPE) excitation with time-correlated single-photon counting techniques to determine time-resolved fluorescence intensity and anisotropy decays of the wild-type Green Fluorescent Protein (GFP) and two red-shifted mutants, S65T-GFP and RSGFP. WT-GFP and S65T-GFP exhibited a predominant approximately 3 ns monoexponential fluorescence decay, whereas for RSGFP the main lifetimes were approximately 1.1 ns (main component) and approximately 3.3 ns. The anisotr...

  18. Fluorescent-magnetic Janus particles prepared via seed emulsion polymerization.

    Science.gov (United States)

    Kaewsaneha, Chariya; Bitar, Ahmad; Tangboriboonrat, Pramuan; Polpanich, Duangporn; Elaissari, Abdelhamid

    2014-06-15

    Anisotropic polymeric colloidal or Janus particles possessing simultaneous magnetic and fluorescent properties were successfully prepared via the swelling-diffusion or the in situ emulsion polymerization method. In the swelling-diffusion process, magnetic emulsions (an organic ferrofluid dispersed in aqueous medium) were synthesized and used for seeds of submicron magnetic Janus particles. After swelling the anisotropic particles obtained by 1-pyrene-carboxaldehyde fluorescent dye dissolved in tetrahydrofuran, well-defined fluorescent-magnetic Janus particles were produced. In the in situ emulsion polymerization, styrene monomer mixed with fluorescent dye monomers, i.e., 1-pyrenylmethyl methacrylate (PyMMA) or fluorescein dimethacrylate (FDMA), and an oil-soluble initiator (2,2'-azobis(2-isobutyronitrile)) were emulsified in the presence of magnetic seed emulsions. The confocal microscopic images showed the fluorescent-magnetic Janus particles with high fluorescent intensity when a fluorescent crosslinker monomer FDMA was employed.

  19. Carboxyfluorescein Diacetate Succinimidyl Ester Fluorescent Dye for Cell Labeling

    Institute of Scientific and Technical Information of China (English)

    Xiao-Qi WANG; Xiu-Mei DUAN; Li-Hua LIU; Yan-Qiu FANG; Yan TAN

    2005-01-01

    Our objective was to study the properties of the carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) and the methodology of cell labeling using CFDA-SE fluorescent dye. First, we analyzed the kinetics of CFDA-SE fluorescent dye intensity over time. Second, we determined the optimal concentration of CFDA-SE fluorescent dye for cell labeling. Third, we tested the toxicity of CFDA-SE fluorescent dye on labeled cells. Finally, we determined the optimal staining time of CFDA-SE fluorescent dye for cell labeling.The results show that the optimal concentration of CFDA-SE fluorescent dye for cell labeling varies according to different cell types. CFDA-SE fluorescent dye is non-toxic to cells as the cell death rate caused by CFDASE labeling is below 5%. The optimal cell labeling time was determined to be 8 min of incubation with CFDA-SE fluorescent dye. We concluded that the advantages of using CFDA-SE fluorescent dye for cell labeling are as follows: (1) the binding of CFDA-SE fluorescent dye to cells is stable; (2) CFDA-SE fluorescent dye is not toxic and does not modify the viability of labeled cells; and (3) CFDA-SE fluorescent dye is a suitable fluorochrome for cell labeling.

  20. 基于GSH-CdTe/CdS量子点的荧光变化研究hsDNA与盐酸洛美沙星-Cu(Ⅱ)配合物的相互作用%Interaction of Herring Sperm DNA with Lomefloxacin Hydrochloride-Cu(Ⅱ) Based on Changes in the Fluorescence Intensity of GSH-CdTe/CdS Quantum Dots

    Institute of Scientific and Technical Information of China (English)

    沈益忠; 刘绍璞; 殷鹏飞; 何佑秋

    2013-01-01

    Glutathione (GSH)-capped CdTe/CdS quantum dots(GSH-CdTe/CdS QDs) were synthesized in aqueous solution.The particle sizes and morphological characteristics of GSH-CdTe/CdS QDs were investigated by transmission electron microscopy(TEM).The results exhibits that the particle size of as-prepared QDs has a narrow size distribution and good dispersivity.In Tris-HCl buffer medium (pH =7.6),lomefloxacin hydrochloride-Cu (Ⅱ) coordination compound(LMFH-Cu2+) was adsorbed to the surfaces of GSH-CdTe/CdS QDs through electrostatic attraction and formed ground state complex,which resulted in the quenching of the fluorescence of GSH-CdTe/CdS QDs.Adding herring sperm DNA (hsDNA) to GSH-CdTe/CdS QDs-LMFH-Cu(Ⅱ) system led to the fluorescence intensity of GSH-CdTe/CdS QDs recover,which can be explained by that the addition of hsDNA to the system induced LMFH-Cu(Ⅱ) to dissociate from the surface of GSH-CdTe/CdS QDs and embed into its double helix structure.According to the fluorescence quenching and restoration for GSH-CdTe/CdS QDs,fluorescence reversible control of GSH-CdTe/CdS QDs was realized.Compared with the interaction between GSH-CdTe/CdS QDs and LMFH,the interaction of GSH-CdTe/CdS QDs-LMFH-Cu (Ⅱ)-hsDNA was studied by fluorescence (FL),resonance Rayleigh scattering(RRS) and ultraviolet-visible absorption (UV-Vis) spectra.Meanwhile,the interaction mechanism was discussed and corresponding model of interaction was built.%采用水相法合成了谷胱甘肽(GSH)修饰的CdTe/CdS量子点(GSH-CdTe/CdS QDs).透射电子显微镜表征结果表明,GSH-CdTe/CdS QDs的粒径分布均匀,分散性好.在Tris-HCl(pH=7.6)缓冲液中,由于静电引力作用,带正电的盐酸洛美沙星(LMFH)-Cu(Ⅱ)配合物[LMFH-Cu(Ⅱ)]吸附到带负电的GSH-CdTe/CdSQDs表面形成基态复合物,导致GSH-CdTe/CdS QDs的荧光猝灭.随后,向GSH-CdTe/CdS QDs-LMFH-Ca(Ⅱ)配合物体系中加入鲱鱼精DNA(hsDNA),hsDNA可诱导LMFH-Cu(Ⅱ)配合物从GSH-CdTe/CdS QDs表面脱落而嵌

  1. Effects of light intensity, temperature and salinity on newborn branches of Sargassum thunbergii evaluated with chlorophyll fluorescence assay%利用叶绿素荧光技术揭示光照、温度和盐度对鼠尾藻嫩芽的影响

    Institute of Scientific and Technical Information of China (English)

    梁洲瑞; 王飞久; 孙修涛; 汪文俊; 丁昌玲; 李涛

    2011-01-01

    The effects of different light intensity (25~220 μmol/(m2·s)), temperatures (5~34 ) and different ℃ erent salinities (0~60) on chlorophyll fluorescence parameters of newborn branches of Sargassum thunbergii were studies in this paper. The optimal chlorophyll fluorescence quantum yield of photosystem II (Fv/Fm), maximum relative electron transport rate (RrET,max) and initial slope of rapid light curve were determined. The results show: (1) The newborn branch of S. thunbergii suffered from high intensity light and high temperature stress easily and the Fv/Fm decreased significantly under high intensity light. The photosynthesis was remarkably affected when the temperature was higher than 30℃. (2) One hour treatment at 5℃ or nine-hour treatment at 0~60 salinity affected the photosystem II obviously. However, the chlorophyll fluorescence parameters could nearly recover to normal level after 24 hours under standard culture condition except those in 60 salinity group. The preliminary analysis of the newborn branches of S. thunbergii about resistance physiology could provide reference for the artificial cultivation of S. thunbergii.%采用鼠尾藻嫩芽为实验材料, 研究了不同光强(25~220 μmol/(m2·s))、温度(5~34℃)和盐度(0~60)对其叶绿素荧光参数的影响。测定的叶绿素荧光参数包括光系统II 最大荧光产量(Fv/Fm)、最大潜在相对电子传递速率(RrET,max)、快速光曲线的初始斜率等。结果表明: (1)鼠尾藻嫩芽易受强光、高温胁迫, 在强光照射下, 其Fv/Fm值明显降低, 温度大于30℃时对其光合作用有显著影响; (2)5℃低温处理1 h 或0~60 盐度处理9 h 对光系统Ⅱ均有明显影响, 但24 h 恢复后, 除了盐度60 组, 其他组的叶绿素荧光参数均可基本恢复正常。初步分析了鼠尾藻嫩芽的抗逆生理, 可为鼠尾藻的人工栽培提供参考。

  2. Construction of In Vivo Fluorescent Imaging of Echinococcus granulosus in a Mouse Model

    Science.gov (United States)

    Wang, Sibo; Yang, Tao; Zhang, Xuyong; Xia, Jie; Guo, Jun; Wang, Xiaoyi; Hou, Jixue; Zhang, Hongwei; Chen, Xueling; Wu, Xiangwei

    2016-01-01

    Human hydatid disease (cystic echinococcosis, CE) is a chronic parasitic infection caused by the larval stage of the cestode Echinococcus granulosus. As the disease mainly affects the liver, approximately 70% of all identified CE cases are detected in this organ. Optical molecular imaging (OMI), a noninvasive imaging technique, has never been used in vivo with the specific molecular markers of CE. Thus, we aimed to construct an in vivo fluorescent imaging mouse model of CE to locate and quantify the presence of the parasites within the liver noninvasively. Drug-treated protoscolices were monitored after marking by JC-1 dye in in vitro and in vivo studies. This work describes for the first time the successful construction of an in vivo model of E. granulosus in a small living experimental animal to achieve dynamic monitoring and observation of multiple time points of the infection course. Using this model, we quantified and analyzed labeled protoscolices based on the intensities of their red and green fluorescence. Interestingly, the ratio of red to green fluorescence intensity not only revealed the location of protoscolices but also determined the viability of the parasites in vivo and in vivo tests. The noninvasive imaging model proposed in this work will be further studied for long-term detection and observation and may potentially be widely utilized in susceptibility testing and therapeutic effect evaluation. PMID:27417083

  3. Construction of In Vivo Fluorescent Imaging of Echinococcus granulosus in a Mouse Model.

    Science.gov (United States)

    Wang, Sibo; Yang, Tao; Zhang, Xuyong; Xia, Jie; Guo, Jun; Wang, Xiaoyi; Hou, Jixue; Zhang, Hongwei; Chen, Xueling; Wu, Xiangwei

    2016-06-01

    Human hydatid disease (cystic echinococcosis, CE) is a chronic parasitic infection caused by the larval stage of the cestode Echinococcus granulosus. As the disease mainly affects the liver, approximately 70% of all identified CE cases are detected in this organ. Optical molecular imaging (OMI), a noninvasive imaging technique, has never been used in vivo with the specific molecular markers of CE. Thus, we aimed to construct an in vivo fluorescent imaging mouse model of CE to locate and quantify the presence of the parasites within the liver noninvasively. Drug-treated protoscolices were monitored after marking by JC-1 dye in in vitro and in vivo studies. This work describes for the first time the successful construction of an in vivo model of E. granulosus in a small living experimental animal to achieve dynamic monitoring and observation of multiple time points of the infection course. Using this model, we quantified and analyzed labeled protoscolices based on the intensities of their red and green fluorescence. Interestingly, the ratio of red to green fluorescence intensity not only revealed the location of protoscolices but also determined the viability of the parasites in vivo and in vivo tests. The noninvasive imaging model proposed in this work will be further studied for long-term detection and observation and may potentially be widely utilized in susceptibility testing and therapeutic effect evaluation.

  4. Dynamic noninvasive monitoring of renal function in vivo by fluorescence lifetime imaging

    Science.gov (United States)

    Goiffon, Reece J.; Akers, Walter J.; Berezin, Mikhail Y.; Lee, Hyeran; Achilefu, Samuel

    2009-03-01

    Kidneys normally filter the blood of excess salts and metabolic products, such as urea, while retaining plasma proteins. In diseases such as multiple myeloma and diabetes mellitus, the renal function is compromised and protein escapes into the urine. In this study, we present the use of fluorescence lifetime imaging (FLI) to image excess serum protein in urine (proteinuria). The near-infrared fluorescent dye LS-288 has distinct lifetimes when bound to protein versus free in solution, providing contrast between the protein-rich viscera and the mostly protein-free bladder. FLI with LS-288 in mice revealed that fluorescence lifetime (FLT) differences in the bladder relative to surrounding tissues was due to the fractional contributions of the bound and unbound dye molecules. The FLT of LS-288 decreased in the case of proteinuria while fluorescence intensity was unchanged. The results show that FLI can be useful for the dynamic imaging of protein-losing nephropathy due to diabetes mellitus and other renal diseases and suggest the potential use of the FLI to distinguish tumors from fluid-filled cysts in the body.

  5. Gene transfer and expression of enhanced green fluorescent protein in variant HT-29c cells

    Institute of Scientific and Technical Information of China (English)

    Min Wang; Lars Boenicke; Bradley D. Howard; Ilka Vogel; Hoiger Kalthoff

    2003-01-01

    AIM: To study the expression of enhanced green fluorescent protein (EGFP) gene in retrovirally transduced variant HT29 cells.METHODS: The retroviral vector prkat EGFP/neo was constructed and transfected into the 293T cell using a standard calcium phosphate precipitation method. HT-29c cells (selected from HT-29 cells) were transduced by a retroviral vector encoding the GEFP gene. The fluorescence intensity of colorectal carcinoma HT-29c cells after transduced with the EGFP bearing retrovirus was visualized using fluorescence microscope and fluorescence activated cell sorter (FACS) analysis. Multiple biological behaviors of transduced cells such as the proliferating potential and the expression of various antigens were comparatively analyzed between untransduced and transduced cells in vitro. EGFP expression of the fresh tumor tissue was assessed in vivo.RESULTS: After transduced, HT-29c cells displayed a stable and long-term EGFP expression under the nonselective conditionsin vitro. After cells were successively cultured to passage 50 in vitro, EGFP expression was still at a high level. Their biological behaviors, such as expression of tumor antigens, proliferation rate and aggregation capability were not different compared to untransduced parental cells in vitro. In subcutaneous tumors, EGFP was stable and highly expressed.CONCLUSION: An EGFP expressing retroviral vector was used to transduce HT-29c cells. The transduced cells show a stable and long-term EGFP expression in vitro and in vivo.These cells with EGFP are a valuable tool forin vivo research of tumor metastatic spread.

  6. Quantitative imaging with fluorescent biosensors.

    Science.gov (United States)

    Okumoto, Sakiko; Jones, Alexander; Frommer, Wolf B

    2012-01-01

    Molecular activities are highly dynamic and can occur locally in subcellular domains or compartments. Neighboring cells in the same tissue can exist in different states. Therefore, quantitative information on the cellular and subcellular dynamics of ions, signaling molecules, and metabolites is critical for functional understanding of organisms. Mass spectrometry is generally used for monitoring ions and metabolites; however, its temporal and spatial resolution are limited. Fluorescent proteins have revolutionized many areas of biology-e.g., fluorescent proteins can report on gene expression or protein localization in real time-yet promoter-based reporters are often slow to report physiologically relevant changes such as calcium oscillations. Therefore, novel tools are required that can be deployed in specific cells and targeted to subcellular compartments in order to quantify target molecule dynamics directly. We require tools that can measure enzyme activities, protein dynamics, and biophysical processes (e.g., membrane potential or molecular tension) with subcellular resolution. Today, we have an extensive suite of tools at our disposal to address these challenges, including translocation sensors, fluorescence-intensity sensors, and Förster resonance energy transfer sensors. This review summarizes sensor design principles, provides a database of sensors for more than 70 different analytes/processes, and gives examples of applications in quantitative live cell imaging.

  7. Interconversion of Anthozoa GFP-like fluorescent and non-fluorescent proteins by mutagenesis

    Directory of Open Access Journals (Sweden)

    Mudrik Nikolay N

    2002-04-01

    Full Text Available Abstract Background Within the family of green fluorescent protein (GFP homologs, one can mark two main groups, specifically, fluorescent proteins (FPs and non-fluorescent or chromoproteins (CPs. Structural background of differences between FPs and CPs are poorly understood to date. Results Here, we applied site-directed and random mutagenesis in order to to transform CP into FP and vice versa. A purple chromoprotein asCP (asFP595 from Anemonia sulcata and a red fluorescent protein DsRed from Discosoma sp. were selected as representatives of CPs and FPs, respectively. For asCP, some substitutions at positions 148 and 165 (numbering in accordance to GFP were found to dramatically increase quantum yield of red fluorescence. For DsRed, substitutions at positions 148, 165, 167, and 203 significantly decreased fluorescence intensity, so that the spectral characteristics of these mutants became more close to those of CPs. Finally, a practically non-fluorescent mutant DsRed-NF was generated. This mutant carried four amino acid substitutions, specifically, S148C, I165N, K167M, and S203A. DsRed-NF possessed a high extinction coefficient and an extremely low quantum yield ( Conclusions We located a novel point in asCP sequence (position 165 mutations at which can result in red fluorescence appearance. Probably, this finding could be applied onto other CPs to generate red and far-red fluorescent mutants. A possibility to transform an FP into CP was demonstrated. Key role of residues adjacent to chromophore's phenolic ring in fluorescent/non-fluorescent states determination was revealed.

  8. Quantification of Coffea arabica and Coffea canephora var. robusta concentration in blends by means of synchronous fluorescence and UV-Vis spectroscopies.

    Science.gov (United States)

    Dankowska, A; Domagała, A; Kowalewski, W

    2017-09-01

    The potential of fluorescence, UV-Vis spectroscopies as well as the low- and mid-level data fusion of both spectroscopies for the quantification of concentrations of roasted Coffea arabica and Coffea canephora var. robusta in coffee blends was investigated. Principal component analysis was used to reduce data multidimensionality. To calculate the level of undeclared addition, multiple linear regression (PCA-MLR) models were used with lowest root mean square error of calibration (RMSEC) of 3.6% and root mean square error of cross-validation (RMSECV) of 7.9%. LDA analysis was applied to fluorescence intensities and UV spectra of Coffea arabica, canephora samples, and their mixtures in order to examine classification ability. The best performance of PCA-LDA analysis was observed for data fusion of UV and fluorescence intensity measurements at wavelength interval of 60nm. LDA showed that data fusion can achieve over 96% of correct classifications (sensitivity) in the test set and 100% of correct classifications in the training set, with low-level data fusion. The corresponding results for individual spectroscopies ranged from 90% (UV-Vis spectroscopy) to 77% (synchronous fluorescence) in the test set, and from 93% to 97% in the training set. The results demonstrate that fluorescence, UV, and visible spectroscopies complement each other, giving a complementary effect for the quantification of roasted Coffea arabica and Coffea canephora var. robusta concentration in blends. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Intensity-intensity and intensity-amplitude correlation of microwave photons from a superconducting artificial atom

    Science.gov (United States)

    Wang, Fei; Feng, Xunli; Oh, C. H.

    2016-10-01

    We investigate the dynamics of the microwave-frequency nonclassical correlations in a three-level Δ -configuration artificial atom, which is realized by superconducting quantum circuits. The intensity-intensity correlation and intensity field are strongly dependent on the relative phase Φ of the driven fields. It is found that two interference loops are formed in the dressed state picture at Φ =0 or π, which are responsible for the generation of nonclassical microwave photons. When the phase is changed into Φ =π /2 or 3π /2 , the temporal correlation functions exhibit different oscillating behaviors. The phase-sensitive nonclassical correlations of fluorescence photons may find practical application in the design of all-optical switches and quantum information processing.

  10. Engineering color variants of green fluorescent protein (GFP) for thermostability, pH-sensitivity, and improved folding kinetics.

    Science.gov (United States)

    Aliye, Naser; Fabbretti, Attilio; Lupidi, Giulio; Tsekoa, Tsepo; Spurio, Roberto

    2015-02-01

    A number of studies have been conducted to improve chromophore maturation, folding kinetics, thermostability, and other traits of green fluorescent protein (GFP). However, no specific work aimed at improving the thermostability of the yellow fluorescent protein (YFP) and of the pH-sensitive, yet thermostable color variants of GFP has so far been done. The protein variants reported in this study were improved through rational multiple site-directed mutagenesis of GFP (ASV) by introducing up to ten point mutations including the mutations near and at the chromophore region. Therefore, we report the development and characterization of fast folder and thermo-tolerant green variant (FF-GFP), and a fast folder thermostable yellow fluorescent protein (FFTS-YFP) endowed with remarkably improved thermostability and folding kinetics. We demonstrate that the fluorescence intensity of this yellow variant is not affected by heating at 75 °C. Moreover, we have developed a pH-unresponsive cyan variant AcS-CFP, which has potential use as part of in vivo imaging irrespective of intracellular pH. The combined improved properties make these fluorescent variants ideal tools to study protein expression and function under different pH environments, in mesophiles and thermophiles. Furthermore, coupling of the FFTS-YFP and AcS-CFP could potentially serve as an ideal tool to perform functional analysis of live cells by multicolor labeling.

  11. Fabrication of fluorescent chitosan-containing microcapsules

    Directory of Open Access Journals (Sweden)

    Zhang R.

    2013-08-01

    Full Text Available Intense emission peaks of Eu(DBM3Phen (DBM and Phen are dibenzoylmethane and 1,10-phenanthroline, respectively in the microcapsules containing molecules of quaternary ammonium chitosan (QACS and sodium alginate are observed. The microcapsules are assembled by using CaCO3 particles as template cores by the layer-by-layer (LbL technique. Observation of microcapsules by the fluorescence mode and the transmission mode in the confocal laser scanning microscopy shows that the microcapsules are intact after core decomposition. Fluorescence under ultraviolet irradiation comes directly from the Eu(DBM3Phen. Homogeneous assembly of Eu(DBM3Phen can be deduced due to the homogeneous fluorescence of the microcapsules in the fluorescence micrographs. The microcapsules show adherence to solid substrates due to large quantities of hydroxyl groups of QACS. AFM measurements of dried hollow microcapsules with only 4 bilayers of (CS/SA fabricated with Eu(DBM3Phen show the intact shell with a thickness of 3.0 nm. Regarding the biocompatible natural polysaccharides and the intense fluorescence emission, the microcapsules in this work might be of great importance in potential application in drug delivery and bioassay.

  12. PHOTODYNAMIC DIAGNOSIS AND FLUORESCENCE SPECTROSCOPY IN SUPERFICIAL BLADDER CANCER

    Directory of Open Access Journals (Sweden)

    I. G. Rusakov

    2009-01-01

    Full Text Available A comprehensive fluorescence technique has been developed to study the urinary bladder mucosa in patients with superficial bladder cancer (BC, by using alasense, white light cystoscopy, fluorescence cytoscopy, and local fluorescence spectroscopy in vivo. Quantification of urothelium fluorescence in the red emission foci of 5-ALA-induced protophorphyrin, with the local autofluorescence intensity being borne in mind, has been shown to increase the specificity of photodynamic diagnosis of superficial BC from 70 to 85% (p ≤ 0.05 and the total accuracy of the technique from 80 to 86%.  

  13. Lymphocyte fluorescent profiles (LFP): a possible screening technique in neoplasia.

    Science.gov (United States)

    Love, L A; Metcalf, N F; Metcalf, W K; Sharp, J G

    1979-05-01

    A technique involving fluorescent protein staining and microfluorometry has been developed for measuring the lymphocyte fluorescent profile (LFP) of peripheral blood lymphocytes. In contrast to normal humans who display a regular bell-shaped curve, the profile from patients with cancer is irregular, showing a bimodal distribution of fluorescence, with a significant population of cells fluorescing at a higher relative intensity. It is suggested that this elevation in protein concentration is due to an immune response to the presence of a neoplasm, and thus this technique may prove to be a useful indicator of malignancy.

  14. Intense electron and ion beams

    CERN Document Server

    Molokovsky, Sergey Ivanovich

    2005-01-01

    Intense Ion and Electron Beams treats intense charged-particle beams used in vacuum tubes, particle beam technology and experimental installations such as free electron lasers and accelerators. It addresses, among other things, the physics and basic theory of intense charged-particle beams; computation and design of charged-particle guns and focusing systems; multiple-beam charged-particle systems; and experimental methods for investigating intense particle beams. The coverage is carefully balanced between the physics of intense charged-particle beams and the design of optical systems for their formation and focusing. It can be recommended to all scientists studying or applying vacuum electronics and charged-particle beam technology, including students, engineers and researchers.

  15. Functionalization of embedded thiol-ene waveguides for evanescent wave induced fluorescence detection in a microfluidic device

    DEFF Research Database (Denmark)

    Feidenhans'l, Nikolaj Agentoft; Jensen, Thomas Glasdam; Lafleur, Josiane P.;

    2013-01-01

    functionalized with biotin using photografting. The biotin was used for immobilization of fluorescently labelled streptavidin, and experiments revealed a linear correlation between streptavidin concentration and fluorescent intensity. To further demonstrate the attractiveness of using thiol−ene for optofluidic...

  16. The first bifluoride sensor based on fluorescent enhancement.

    Science.gov (United States)

    Dutta, Kaku; Deka, Ramesh Ch; Das, Diganta Kumar

    2013-07-01

    The first fluorescent sensor for HF2(-) anion, N(1), N(3)-di(naphthalene-1-yl)isophthalamide (L) has been derived from α-Napthylamine and isopthaloyl chloride. In 1:1 (v/v) DMSO:H2O, L exhibits high selectivity towards HF2(-) anion with a 4-fold enhancement in fluorescent intensity. Very little enhancement in fluorescence intensity is observed for F(-), Cl(-), Br(-), I(-), SCN(-), PO4(3-), SO4(2-), and CH3COO(-) anions. The stoichiometry interaction between L and HF2 (-) is found to be 1:1 from fluorescence and UV/Visible spectral data. DFT calculation shows that binding between HF2(-) and L is 1:1 and increases the relative planarity between the two naphthyl rings causing fluorescence enhancement. A shift of 0.080 V in oxidation potential of L is observed on interaction with HF2(-) by cyclic voltammetry and square wave voltammetry.

  17. Tailoring cyanine dark states for improved optically modulated fluorescence recovery.

    Science.gov (United States)

    Mahoney, Daniel P; Owens, Eric A; Fan, Chaoyang; Hsiang, Jung-Cheng; Henary, Maged M; Dickson, Robert M

    2015-04-02

    Cyanine dyes are well-known for their bright fluorescence and utility in biological imaging. However, cyanines also readily photoisomerize to produce nonemissive dark states. Co-illumination with a secondary, red-shifted light source on-resonance with the longer wavelength absorbing dark state reverses the photoisomerization and returns the cyanine dye to the fluorescent manifold, increasing steady-state fluorescence intensity. Modulation of this secondary light source dynamically alters emission intensity, drastically improving detection sensitivity and facilitating fluorescence signals to be recovered from an otherwise overwhelming background. Red and near-IR emitting cyanine derivatives have been synthesized with varying alkyl chain lengths and halogen substituents to alter dual-laser fluorescence enhancement. Photophysical properties and enhancement with dual laser modulation were coupled with density functional calculations to characterize substituent effects on dark state photophysics, potentially improving detection in high background biological environments.

  18. SIMULTANEOUS MEASUREMENT OF CIRCULAR DICHROISM AND FLUORESCENCE POLARIZATION ANISOTROPY.

    Energy Technology Data Exchange (ETDEWEB)

    SUTHERLAND,J.C.

    2002-01-19

    Circular dichroism and fluorescence polarization anisotropy are important tools for characterizing biomolecular systems. Both are used extensively in kinetic experiments involving stopped- or continuous flow systems as well as titrations and steady-state spectroscopy. This paper presents the theory for determining circular dichroism and fluorescence polarization anisotropy simultaneously, thus insuring the two parameters are recorded under exactly the same conditions and at exactly the same time in kinetic experiments. The approach to measuring circular dichroism is that used in almost all conventional dichrographs. Two arrangements for measuring fluorescence polarization anisotropy are described. One uses a single fluorescence detector and signal processing with a lock-in amplifier that is similar to the measurement of circular dichroism. The second approach uses classic ''T'' format detection optics, and thus can be used with conventional photon-counting detection electronics. Simple extensions permit the simultaneous measurement of the absorption and excitation intensity corrected fluorescence intensity.

  19. PNA-induced assembly of fluorescent proteins using DNA as a framework.

    Science.gov (United States)

    Gholami, Zahra; Brunsveld, Luc; Hanley, Quentin

    2013-08-21

    Controlled alignment of proteins on molecular frameworks requires the development of facile and orthogonal chemical approaches and molecular scaffolds. In this work, protein-PNA conjugates are brought forward as new chemical components allowing efficient assembly and alignment on DNA scaffolds. Site-selective monomeric teal fluorescent protein (mTFP)-peptide nucleic acid (PNA) (mTFP-PNA) conjugation was achieved by covalent linkage of the PNA to the protein through expressed protein ligation (EPL). A DNA beacon, with 6-Fam and Dabcyl at its ends, acts as a framework to create an assembled hetero-FRET system with the mTFP-PNA conjugate. Using fluorescence intensity, frequency domain lifetime measurements, and anisotropy measurements, the system was shown to produce FRET as indicated by decreased donor intensity, decreased donor lifetime, and increased donor anisotropy. Extension of the DNA scaffold allowed for the assembly of multiple mTFP-PNA constructs. Efficient formation of protein dimers and oligomers on the DNA-PNA frameworks could be shown, as visualized via size exclusion chromatography (SEC) and electrophoresis (SDS-PAGE). Assembly of multiple proteins in a row induced homo-FRET for the mTFP-PNA's assembled on the DNA scaffolds. The oligonucleotide framework allows an induced and controllable assembly of proteins by fusing them to PNAs directed to align on DNA scaffolds.

  20. Synthesis of 2,3-dimorpholino-6-aminoquinoxaline derivatives and application to a new intramolecular fluorescent probe

    Energy Technology Data Exchange (ETDEWEB)

    Matsumura, Yuriko [Department of Materials and Life Science, Faculty of Science and Technology, Seikei University, 3-3-1 Kitamachi, Kichijoji, Musashino-shi, Tokyo 180-8633 (Japan)], E-mail: matsumura@st.seikei.ac.jp; Katoh, Akira [Department of Materials and Life Science, Faculty of Science and Technology, Seikei University, 3-3-1 Kitamachi, Kichijoji, Musashino-shi, Tokyo 180-8633 (Japan)], E-mail: katoh@st.seikei.ac.jp

    2008-04-15

    Two fluorescent monomers having a quinoxaline skeleton, N-(2,3-dimorpholinoquinoxalin-6-yl)acrylamide (QxA) and N-(1-(2,3-dimorpholinoquinoxalin-6-ylamino)prop-2-yl)methacrylamide (QxAlaMA), were synthesized. Thermo-responsive copolymers of N-isopropylacrylamide (NIPAM) and a small amount of a fluorescent monomer were synthesized and their fluorescence properties investigated. The fluorescent monomers showed intense solvatochromism in their fluorescence. The wavelength at the maximum fluorescence intensity of the QxAlaMA-labeled PNIPAM dramatically blue-shifted and the fluorescence intensity of the QxA-labeled PNIPAM significantly increased around the transition temperature. It was found that these fluorescent dyes can sense and report the thermo-responsive behavior of the PNIPAM in water. Both QxAlaMA and QxA were demonstrated to be applicable to new intramolecular fluorescent probes.

  1. A robust co-localisation measurement utilising z-stack image intensity similarities for biological studies.

    Directory of Open Access Journals (Sweden)

    Yinhai Wang

    Full Text Available BACKGROUND: Co-localisation is a widely used measurement in immunohistochemical analysis to determine if fluorescently labelled biological entities, such as cells, proteins or molecules share a same location. However the measurement of co-localisation is challenging due to the complex nature of such fluorescent images, especially when multiple focal planes are captured. The current state-of-art co-localisation measurements of 3-dimensional (3D image stacks are biased by noise and cross-overs from non-consecutive planes. METHOD: In this study, we have developed Co-localisation Intensity Coefficients (CICs and Co-localisation Binary Coefficients (CBCs, which uses rich z-stack data from neighbouring focal planes to identify similarities between image intensities of two and potentially more fluorescently-labelled biological entities. This was developed using z-stack images from murine organotypic slice cultures from central nervous system tissue, and two sets of pseudo-data. A large amount of non-specific cross-over situations are excluded using this method. This proposed method is also proven to be robust in recognising co-localisations even when images are polluted with a range of noises. RESULTS: The proposed CBCs and CICs produce robust co-localisation measurements which are easy to interpret, resilient to noise and capable of removing a large amount of false positivity, such as non-specific cross-overs. Performance of this method of measurement is significantly more accurate than existing measurements, as determined statistically using pseudo datasets of known values. This method provides an important and reliable tool for fluorescent 3D neurobiological studies, and will benefit other biological studies which measure fluorescence co-localisation in 3D.

  2. Fluorescent N-Doped Carbon Dots as in Vitro and in Vivo Nanothermometer.

    Science.gov (United States)

    Yang, Yanmei; Kong, Weiqian; Li, Hao; Liu, Juan; Yang, Manman; Huang, Hui; Liu, Yang; Wang, Zhongyang; Wang, Zhiqiang; Sham, Tsun-Kong; Zhong, Jun; Wang, Chao; Liu, Zhuang; Lee, Shuit-Tong; Kang, Zhenhui

    2015-12-16

    The fluorescent N-doped carbon dots (N-CDs) obtained from C3N4 emit strong blue fluorescence, which is stable with different ionic strengths and time. The fluorescence intensity of N-CDs decreases with the temperature increasing, while it can recover to the initial one with the temperature decreasing. It is an accurate linear response of fluorescence intensity to temperature, which may be attributed to the synergistic effect of abundant oxygen-containing functional groups and hydrogen bonds. Further experiments also demonstrate that N-CDs can serve as effective in vitro and in vivo fluorescence-based nanothermometer.

  3. Note: Rapid measurement of fluorescence lifetimes using SiPM detection and waveform sampling

    Science.gov (United States)

    Tsai, H.-M.; Souris, J. S.; Kim, H.-J.; Cheng, S.-H.; Chen, L.; Lo, L.-W.; Chen, C.-T.; Kao, C.-M.

    2017-09-01

    In fluorescence spectroscopy and imaging, fluorescence lifetime measurement—assessing the average time fluorophores spend in their excited state before returning to their ground state—offers a number of advantages over quantifying fluorescence intensities that include resistance to photo-bleaching and independence from fluorophore concentration, excitation intensity, and measurement methodology. Despite growing interest, fluorescence lifetime techniques frequently mandate relatively complex instrumentation, slow data acquisition rates, and significant data analyses. In this work, we demonstrate the feasibility of measuring fluorescence lifetimes using off-the-shelf analog silicon photomultipliers and switched-capacitor array waveform sampling techniques, with precision matching that of much larger and more elaborate commercial instruments.

  4. Multiple Pregnancy

    Science.gov (United States)

    ... Education & Events Advocacy For Patients About ACOG Multiple Pregnancy Home For Patients Search FAQs Multiple Pregnancy Page ... Multiple Pregnancy FAQ188, July 2015 PDF Format Multiple Pregnancy Pregnancy How does multiple pregnancy occur? What are ...

  5. A highly selective fluorescent sensor for glucosamine.

    Science.gov (United States)

    Tran, Tam Minh; Alan, Yuksel; Glass, Timothy Edward

    2015-05-07

    A new fluorescent chemical sensor for glucosamine is reported. The sensor is based on a boronic acid-containing coumarin aldehyde and shows excellent selectivity for glucosamine by forming a boronic ester with the sugar diol as well as an iminium ion with the amine group of glucosamine. The sensor successfully discriminates glucosamine over other similar biomolecules in terms of both fluorescence intensity and binding affinity. This method provides a new concept for the design and synthesis of very selective turn-on optical sensors for selective detection of multi-functional biomolecules.

  6. Tomato seeds maturity detection system based on chlorophyll fluorescence

    Science.gov (United States)

    Li, Cuiling; Wang, Xiu; Meng, Zhijun

    2016-10-01

    Chlorophyll fluorescence intensity can be used as seed maturity and quality evaluation indicator. Chlorophyll fluorescence intensity of seed coats is tested to judge the level of chlorophyll content in seeds, and further to judge the maturity and quality of seeds. This research developed a detection system of tomato seeds maturity based on chlorophyll fluorescence spectrum technology, the system included an excitation light source unit, a fluorescent signal acquisition unit and a data processing unit. The excitation light source unit consisted of two high power LEDs, two radiators and two constant current power supplies, and it was designed to excite chlorophyll fluorescence of tomato seeds. The fluorescent signal acquisition unit was made up of a fluorescence spectrometer, an optical fiber, an optical fiber scaffolds and a narrowband filter. The data processing unit mainly included a computer. Tomato fruits of green ripe stage, discoloration stage, firm ripe stage and full ripe stage were harvested, and their seeds were collected directly. In this research, the developed tomato seeds maturity testing system was used to collect fluorescence spectrums of tomato seeds of different maturities. Principal component analysis (PCA) method was utilized to reduce the dimension of spectral data and extract principal components, and PCA was combined with linear discriminant analysis (LDA) to establish discriminant model of tomato seeds maturity, the discriminant accuracy was greater than 90%. Research results show that using chlorophyll fluorescence spectrum technology is feasible for seeds maturity detection, and the developed tomato seeds maturity testing system has high detection accuracy.

  7. Laser-induced fluorescence in diagnosis of dental caries

    Science.gov (United States)

    Drakaki, Eleni A.; Makropoulou, Mersini I.; Khabbaz, Maruan; Serafetinides, Alexandros A.

    2003-09-01

    The autofluorescence spectra of hard dental tissues, both in normal and pathological areas were investigated in this study. The measurements were performed both on the intact hard tissues of the examined teeth, such as enamel, dentine, cementum, and root canal, and on the tissues pathologically affected by caries (superficial, intermediate, and deep). Various laser wavelengths (337 nm, 488 nm, and 514 nm) were used to irradiate the dental surfaces and a computer-controlled spectrograph captured the fluorescent spectra. The emission signals were stored, measured, analyzed and quantified in terms of wavelength distribution and the relative photon intensity. Results indicated that the fluorescent spectra from healthy enamel, dentine, and cementum were almost identical in form, depending on the excitation wavelength. The intact and affected hard tissues were greatly different in the integral fluorescent intensity. Healthy areas were found to produce the most pronounced fluorescent intensity, whereas the carious regions produced the weaker fluorescent intensity. Independently of the laser excitation wavelength, dentin regions were found to produce the most pronounced fluorescent intensity than any other dental component. The fluorescence signal of carious affected dental structure revealed a reed shifted spectral curve, more pronounced after 488 nm excitation. There was a pronounced red shift for deep caries (crown -- root caries), after ultraviolet laser excitation. Excitation with visible wavelengths did not produce such differences between intact and cervical, deep carious affected tissue. Using a monochromatic light source without any light output at the wavelengths of fluorescence, e.g. a laser with the appropriate filters, the difference in fluorescence between intact and carious enamel was generally easy to observe. Finally, we found that the blue line of an argon ion laser is preferable for superficial caries detection, while the ultraviolet emitting nitrogen

  8. Computer generated holography with intensity-graded patterns

    Directory of Open Access Journals (Sweden)

    Rossella Conti

    2016-10-01

    Full Text Available Computer Generated Holography achieves patterned illumination at the sample plane through phase modulation of the laser beam at the objective back aperture. This is obtained by using liquid crystal-based spatial light modulators (LC-SLMs, which modulate the spatial phase of the incident laser beam. A variety of algorithms are employed to calculate the phase modulation masks addressed to the LC-SLM. These algorithms range from simple gratings-and-lenses to generate multiple diffraction-limited spots, to iterative Fourier-transform algorithms capable of generating arbitrary illumination shapes perfectly tailored on the base of the target contour. Applications for holographic light patterning include multi-trap optical tweezers, patterned voltage imaging and optical control of neuronal excitation using uncaging or optogenetics. These past implementations of computer generated holography used binary input profile to generate binary light distribution at the sample plane. Here we demonstrate that using graded input sources, enables generating intensity graded light patterns and extend the range of application of holographic light illumination. At first, we use intensity-graded holograms to compensate for LC-SLM position dependent diffraction efficiency or sample fluorescence inhomogeneity. Finally we show that intensity-graded holography can be used to equalize photo evoked currents from cells expressing different level of chanelrhodopsin2 (ChR2, one of the most commonly used optogenetics light gated channels, taking into account the non-linear dependence of channel opening on incident light.

  9. Saturated excitation of fluorescence to quantify excitation enhancement in aperture antennas.

    Science.gov (United States)

    Aouani, Heykel; Hostein, Richard; Mahboub, Oussama; Devaux, Eloïse; Rigneault, Hervé; Ebbesen, Thomas W; Wenger, Jérôme

    2012-07-30

    Fluorescence spectroscopy is widely used to probe the electromagnetic intensity amplification on optical antennas, yet measuring the excitation intensity amplification is a challenge, as the detected fluorescence signal is an intricate combination of excitation and emission. Here, we describe a novel approach to quantify the electromagnetic amplification in aperture antennas by taking advantage of the intrinsic non linear properties of the fluorescence process. Experimental measurements of the fundamental f and second harmonic 2f amplitudes of the fluorescence signal upon excitation modulation are used to quantify the electromagnetic intensity amplification with plasmonic aperture antennas.

  10. Saturated excitation of Fluorescence to quantify excitation enhancement in aperture antennas

    KAUST Repository

    Aouani, Heykel

    2012-07-23

    Fluorescence spectroscopy is widely used to probe the electromagnetic intensity amplification on optical antennas, yet measuring the excitation intensity amplification is a challenge, as the detected fluorescence signal is an intricate combination of excitation and emission. Here, we describe a novel approach to quantify the electromagnetic amplification in aperture antennas by taking advantage of the intrinsic non linear properties of the fluorescence process. Experimental measurements of the fundamental f and second harmonic 2f amplitudes of the fluorescence signal upon excitation modulation are used to quantify the electromagnetic intensity amplification with plasmonic aperture antennas. © 2012 Optical Society of America.

  11. Normalized fluorescence lifetime imaging for tumor identification and margin delineation

    Science.gov (United States)

    Sherman, Adria J.; Papour, Asael; Bhargava, Siddharth; Taylor, Zach; Grundfest, Warren S.; Stafsudd, Oscar M.

    2013-03-01

    Fluorescence lifetime imaging microscopy (FLIM) is a technique that has been proven to produce quantitative and qualitative differentiation and identification of substances with good specificity and sensitivity based on lifetime extracted information. This technique has shown the ability to also differentiate between a wide range of tissue types to identify malignant from benign tissue in vivo and ex vivo. However, the complexity, long duration and effort required to generate this information has limited the adoption of these techniques in a clinical setting. Our group has developed a time-resolved imaging system (patent pending) that does not require the extraction of lifetimes or use of complex curve fitting algorithms to display the needed information. The technique, entitled Lifetime Fluorescence Imaging (LFI, or NoFYI), converts fluorescence lifetime decay information directly into visual contrast. Initial studies using Fluorescein and Rhodamine-B demonstrated the feasibility of this approach. Subsequent studies demonstrated the ability to separate collagen and elastin powders. The technique uses nanosecond pulsed UV LEDs at 375 nm for average illumination intensities of ~4.5 μW on the tissue surface with detection by a gated CCD camera. To date, we have imaged 11 surgical head and neck squamous cell carcinoma and brain cancer biopsy specimens including 5 normal and 6 malignant samples. Images at multiple wavelengths clearly demonstrate differentiation between benign and malignant tissue, which was later confirmed by histology. Contrast was obtained between fluorophores with 35 μm spatial resolution and an SNR of ~30 dB allowing us to clearly define tumor margins in these highly invasive cancers. This method is capable of providing both anatomical and chemical information for the pathologist and the surgeon. These results suggest that this technology has a possible role in identifying tumors in tissue specimens and detecting tumor margins during procedures.

  12. Multiple sclerosis; Multiple Sklerose

    Energy Technology Data Exchange (ETDEWEB)

    Grunwald, I.Q.; Kuehn, A.L.; Backens, M.; Papanagiotou, P. [Universitaet des Saarlandes, Abteilung fuer Diagnostische und Interventionelle Neuroradiologie, Radiologische Klinik, Homburg/Saar (Germany); Shariat, K. [Universitaet des Saarlandes, Klinik fuer Neurochirurgie, Homburg/Saar (Germany); Kostopoulos, P. [Universitaet des Saarlandes, Klinik fuer Neurologie, Homburg/Saar (Germany)

    2008-06-15

    Multiple sclerosis is the most common chronic inflammatory disease of myelin with interspersed lesions in the white matter of the central nervous system. Magnetic resonance imaging (MRI) plays a key role in the diagnosis and monitoring of white matter diseases. This article focuses on key findings in multiple sclerosis as detected by MRI. (orig.) [German] Die Multiple Sklerose (MS) ist die haeufigste chronisch-entzuendliche Erkrankung des Myelins mit eingesprengten Laesionen im Bereich der weissen Substanz des zentralen Nervensystems. Die Magnetresonanztomographie (MRT) hat bei der Diagnosestellung und Verlaufskontrolle eine Schluesselrolle. Dieser Artikel befasst sich mit Hauptcharakteristika der MR-Bildbebung. (orig.)

  13. Tryptophan content for monitoring breast cancer cell aggressiveness by native fluorescence spectroscopy

    Science.gov (United States)

    Zhang, Lin; Pu, Yang; Xue, Jianpeng; Pratavieira, Sebastião.; Xu, Baogang; Achilefu, Samuel; Alfano, R. R.

    2014-03-01

    This study shows tryptophan as the key native marker in cells to determine the level of aggressive cancer in breast cell lines using native fluorescence spectroscopy. An algorithm based on the ratio of tryptophan fluorescence intensity at 340 nm to intensity at 460 nm is associated with aggressiveness of the cancer cells. The higher the ratio is, the more aggressive the tumor towards metastasis.

  14. Synthesis and fluorescence enhancement behavior of a novel fluorescent aqueous polyurethane emulsion DDAQ-TDI-PU

    Institute of Scientific and Technical Information of China (English)

    Xian Hai Hu; Xing Yuan Zhang; Jia Bing Dai

    2012-01-01

    A novel fluorescent aqueous polyurethane emulsion DDAQ-TDI-PU was synthesized by blocking the anthraquinone moiety of 1,4-diamino-2,3-diphenoxyanthraquinone (DDAQ) into polyurethane chain using 2,4-tolylene diisocyanate (TDI),poly(propylene glycol) and 2,2-dimethylol propionic acid.The chain structure of DDAQ-TDI-PU was confirmed by means of Fourier transform infrared spectroscopy and UV-vis analysis.Comparing to the UV-vis spectrum of DDAQ,DDAQ-TDI-PU showed a hypsochromic shift from the absorption maxima of 518,558,609 nm to 510,548,586 nm,respectively.It was found that the fluorescence intensity of DDAQ-TDI-PU emulsion was enhanced greatly comparing with that of DDAQ.The fluorescence of DDAQ-TDI-PU was very stable not only for the long term storage but also for the fluorescence quencher.

  15. Fluorescent Lamp Replacement Study

    Science.gov (United States)

    2017-07-01

    Light-emitting diode, LED, lighting, fluorescent, waste reduction, energy conservation, net zero , mercury 16. SECURITY CLASSIFICATION OF: 17. LIMITATION...Center (ATC) to assess the benefits of converting fluorescent tube lighting to light-emitting diode (LED) technology. The report documents the waste ...1-15 SECTION 2. SUBTESTS 2.1 HAZARDOUS WASTE REDUCTION

  16. LEDs for fluorescence microscopy

    NARCIS (Netherlands)

    Young, I.T.; Garini, Y.; Dietrich, H.R.C.; Van Oel, W.; Liqui Lung, G.

    2004-01-01

    Traditional light sources for fluorescence microscopy have been mercury lamps, xenon lamps, and lasers. These sources have been essential in the development of fluorescence microscopy but each can have serious disadvantages: lack of near monochromaticity, heat generation, cost, lifetime of the light

  17. Development of Functional Fluorescent Molecular Probes for the Detection of Biological Substances

    Directory of Open Access Journals (Sweden)

    Yoshio Suzuki

    2015-06-01

    Full Text Available This review is confined to sensors that use fluorescence to transmit biochemical information. Fluorescence is, by far, the most frequently exploited phenomenon for chemical sensors and biosensors. Parameters that define the application of such sensors include intensity, decay time, anisotropy, quenching efficiency, and luminescence energy transfer. To achieve selective (biomolecular recognition based on these fluorescence phenomena, various fluorescent elements such as small organic molecules, enzymes, antibodies, and oligonucleotides have been designed and synthesized over the past decades. This review describes the immense variety of fluorescent probes that have been designed for the recognitions of ions, small and large molecules, and their biological applications in terms of intracellular fluorescent imaging techniques.

  18. Multiparameter fluorescence spectroscopic imaging of cell function

    Science.gov (United States)

    Bright, Gary R.

    1994-08-01

    The ability to quantitate physiological parameters in single living cells using fluorescence spectroscopic imaging has expanded our understanding of many cell regulatory processes. Previous studies have focussed on the measurement of single parameters, such as the concentration of calcium, and more recently two parameters, such as calcium and pH using fluorescence ratio imaging. The complexity of the interrelationships among cell biochemical reactions suggests a need to extend the measurement scheme to several parameters. Expansion of the number of parameters involves several complexities associated with fluorescent probe selection and instrumentation design as well as the processing and management of the data. A system has been assembled which provides maximum flexibility in multiparameter fluorescence imaging measurements. The system provides multiple combinations of excitation, dichroic mirror, and emission wavelengths. It has automatic acquisition of any number of parameters. The number of parameters is primarily limited by the selection of fluorescent probes with nonoverlapping spectra. We demonstrate the utility of the system by the coordinated monitoring of stimulated changes in the concentrations of calcium, magnesium, and pH using fluorescence ratio imaging coupled with a conventional transmitted light image of single smooth muscle cells. The results demonstrate coordinated changes in some instances but uncoordinated changes in others.

  19. Measuring Physical Activity Intensity

    Science.gov (United States)

    ... aerobic activity: relative intensity and absolute intensity. Relative Intensity The level of effort required by a person to do an activity. When using relative intensity, people pay attention to how physical activity affects ...

  20. Measuring Physical Activity Intensity

    Medline Plus

    Full Text Available ... aerobic activity: relative intensity and absolute intensity. Relative Intensity The level of effort required by a person to do an activity. When using relative intensity, people pay attention to how physical activity affects ...

  1. Measuring Physical Activity Intensity

    Medline Plus

    Full Text Available ... Compartir For more help with what counts as aerobic activity, watch this video: Windows Media Player, 4: ... ways to understand and measure the intensity of aerobic activity: relative intensity and absolute intensity. Relative Intensity ...

  2. Conditions for NIR fluorescence-guided tumor resectioning in preclinical lung cancer model (Conference Presentation)

    Science.gov (United States)

    Kim, Minji; Quan, Yuhua; Choi, Byeong Hyun; Choi, Yeonho; Kim, Hyun Koo; Kim, Beop-Min

    2016-03-01

    Pulmonary nodule could be identified by intraoperative fluorescence imaging system from systemic injection of indocyanine green (ICG) which achieves enhanced permeability and retention (EPR) effects. This study was performed to evaluate optimal injection time of ICG for detecting cancer during surgery in rabbit lung cancer model. VX2 carcinoma cell was injected in rabbit lung under fluoroscopic computed tomography-guidance. Solitary lung cancer was confirmed on positron emitting tomography with CT (PET/CT) 2 weeks after inoculation. ICG was administered intravenously and fluorescent intensity of lung tumor was measured using the custom-built intraoperative color and fluorescence merged imaging system (ICFIS) for 15 hours. Solitary lung cancer was resected through thoracoscopic version of ICFIS. ICG was observed in all animals. Because Lung has fast blood pulmonary circulation, Fluorescent signal showed maximum intensity earlier than previous studies in other organs. Fluorescent intensity showed maximum intensity within 6-9 hours in rabbit lung cancer. Overall, Fluorescent intensity decreased with increasing time, however, all tumors were detectable using fluorescent images until 12 hours. In conclusion, while there had been studies in other organs showed that optimal injection time was at least 24 hours before operation, this study showed shorter optimal injection time at lung cancer. Since fluorescent signal showed the maximum intensity within 6-9 hours, cancer resection could be performed during this time. This data informed us that optimal injection time of ICG should be evaluated in each different solid organ tumor for fluorescent image guided surgery.

  3. Measurement of protein-like fluorescence in river and waste water using a handheld spectrophotometer.

    Science.gov (United States)

    Baker, Andy; Ward, David; Lieten, Shakti H; Periera, Ryan; Simpson, Ellie C; Slater, Malcolm

    2004-07-01

    Protein-like fluorescence intensity in rivers increases with increasing anthropogenic DOM inputs from sewerage and farm wastes. Here, a portable luminescence spectrophotometer was used to investigate if this technology could be used to provide both field scientists with a rapid pollution monitoring tool and process control engineers with a portable waste water monitoring device, through the measurement of river and waste water tryptophan-like fluorescence from a range of rivers in NE England and from effluents from within two waste water treatment plants. The portable spectrophotometer determined that waste waters and sewerage effluents had the highest tryptophan-like fluorescence intensity, urban streams had an intermediate tryptophan-like fluorescence intensity, and the upstream river samples of good water quality the lowest tryptophan-like fluorescence intensity. Replicate samples demonstrated that fluorescence intensity is reproducible to +/- 20% for low fluorescence, 'clean' river water samples and +/- 5% for urban water and waste waters. Correlations between fluorescence measured by the portable spectrophotometer with a conventional bench machine were 0.91; (Spearman's rho, n = 143), demonstrating that the portable spectrophotometer does correlate with tryptophan-like fluorescence intensity measured using the bench spectrophotometer.

  4. Canopy Level Solar Induced Fluorescence for Vegetation in Controlled Experiments

    Science.gov (United States)

    Middleton, E. M.; Corp, L. A.; Campbell, P. K. Entcheva

    2007-01-01

    Solar induced chlorophyll fluorescence (SIF) was retrieved from high resolution reflectance spectra acquired one meter above saplings of three deciduous tree species during springtime (three weeks after leaf flush) and in late summer when foliage was mature. SIF was determined by application of the Fraunhofer Line Depth (FLD) Principal to above-canopy spectra acquired with an Analytical Spectral Devices (ASD) Fieldspec spectroradiometer (3.2 nm resolution with 1.2 nm sampling interval). SIF retrievals were made at the two atmospheric oxygen (O2) absorption features that occur in the chlorophyll fluorescence (ChlF) region (660 -780 nm). These telluric features are 02V, the broader and deeper feature centered at 760 nm, but located on the shoulder of the far-red ChlF peak at 740 nm; and 023, a narrow feature centered at 688 nm that is positioned near the red ChlF peak at 685 nm. Supporting, coincident leaf level fluorescence, reflectance, photochemical and other measurements were also made. At the leaf level, these measurements included in situ photosynthetic capacity (Pmax) and light adapted total chlorophyll fluorescence (Fs') collected at steady state under high light and controlled chamber conditions (e.g., temperature, PAR, humidity, and COz); optical properties (reflectance, transmittance, absorptance); chlorophyll and carotenoid content; specific leaf mass; carbon (C) and nitrogen (N) content; fluorescence emission spectra at multiple excitation wavelengths; the ChlF contribution to red (R) and far-red (FR) reflectance; fluorescence imagery; and fluorescence excitation-emission matrices (EEMs). The tree species examined were tulip poplar (Liriodendron tulipifera L.), red maple (Acer rubrum L.), and sweetgum (Liquidambar styraczflua L.), and each had been provided four levels of N augmentation (0, 19, 37, and 75 kg Nhectare seasonally) to simulate atmospheric deposition from air pollution. Whole-plant SIF measurements of these species were compared with SIF

  5. The development of attenuation compensation models of fluorescence spectroscopy signals

    Science.gov (United States)

    Dremin, Victor V.; Zherebtsov, Evgeny A.; Rafailov, Ilya E.; Vinokurov, Andrey Y.; Novikova, Irina N.; Zherebtsova, Angelina I.; Litvinova, Karina S.; Dunaev, Andrey V.

    2016-04-01

    This study examines the effect of blood absorption on the endogenous fluorescence signal intensity of biological tissues. Experimental studies were conducted to identify these effects. To register the fluorescence intensity, the fluorescence spectroscopy method was employed. The intensity of the blood flow was measured by laser Doppler flowmetry. We proposed one possible implementation of the Monte Carlo method for the theoretical analysis of the effect of blood on the fluorescence signals. The simulation is constructed as a four-layer skin optical model based on the known optical parameters of the skin with different levels of blood supply. With the help of the simulation, we demonstrate how the level of blood supply can affect the appearance of the fluorescence spectra. In addition, to describe the properties of biological tissue, which may affect the fluorescence spectra, we turned to the method of diffuse reflectance spectroscopy (DRS). Using the spectral data provided by the DRS, the tissue attenuation effect can be extracted and used to correct the fluorescence spectra.

  6. Fluorescence spectroscopy of the retina from scrapie-infected mice.

    Science.gov (United States)

    Bose, Sayantan; Schönenbrücher, Holger; Richt, Jürgen A; Casey, Thomas A; Rasmussen, Mark A; Kehrli, Marcus E; Petrich, Jacob W

    2013-01-01

    Recently, we have proposed that the fluorescence spectra of sheep retina can be well correlated with the presence or absence of scrapie. Scrapie is the most widespread TSE (transmissible spongiform encephalopathy) affecting sheep and goats worldwide. Mice eyes have been previously reported as a model system to study age-related accumulation of lipofuscin, which has been investigated by monitoring the increasing fluorescence with age covering its entire life span. The current work aims at developing mice retina as a convenient model system to diagnose scrapie and other fatal TSE diseases in animals such as sheep and cows. The objective of the research reported here was to determine whether the spectral features are conserved between two different species namely mice and sheep, and whether an appropriate small animal model system could be identified for diagnosis of scrapie based on the fluorescence intensity in retina. The results were consistent with the previous reports on fluorescence studies of healthy and scrapie-infected retina of sheep. The fluorescence from the retinas of scrapie-infected sheep was significantly more intense and showed more heterogeneity than that from the retinas of uninfected mice. Although the structural characteristics of fluorescence spectra of scrapie-infected sheep and mice eyes are slightly different, more importantly, murine retinas reflect the enhancement of fluorescence intensity upon infecting the mice with scrapie, which is consistent with the observations in sheep eyes.

  7. Semi-automated discrimination of retinal pigmented epithelial cells in two-photon fluorescence images of mouse retinas

    Science.gov (United States)

    Alexander, Nathan S.; Palczewska, Grazyna; Palczewski, Krzysztof

    2015-01-01

    Automated image segmentation is a critical step toward achieving a quantitative evaluation of disease states with imaging techniques. Two-photon fluorescence microscopy (TPM) has been employed to visualize the retinal pigmented epithelium (RPE) and provide images indicating the health of the retina. However, segmentation of RPE cells within TPM images is difficult due to small differences in fluorescence intensity between cell borders and cell bodies. Here we present a semi-automated method for segmenting RPE cells that relies upon multiple weak features that differentiate cell borders from the remaining image. These features were scored by a search optimization procedure that built up the cell border in segments around a nucleus of interest. With six images used as a test, our method correctly identified cell borders for 69% of nuclei on average. Performance was strongly dependent upon increasing retinosome content in the RPE. TPM image analysis has the potential of providing improved early quantitative assessments of diseases affecting the RPE. PMID:26309765

  8. Laser induced fluorescence of dental caries

    Science.gov (United States)

    Albin, S.; Byvik, C. E.; Buoncristiani, A. M.

    1988-01-01

    Significant differences between the optical spectra taken from sound regions of teeth and carious regions have been observed. These differences appear both in absorption and in laser induced fluorescence spectra. Excitation by the 488 nm line of an argon ion laser beam showed a peak in the emission intensity around 553 nm for the sound dental material while the emission peak from the carious region was red-shifted by approximately 40 nm. The relative absorption of carious region was significantly higher at 488 nm; however its fluorescence intensity peak was lower by an order of magnitude compared to the sound tooth. Implications of these results for a safe, reliable and early detection of dental caries are discussed.

  9. Laser induced fluorescence of dental caries

    Science.gov (United States)

    Albin, S.; Byvik, C. E.; Buoncristiani, A. M.

    1988-01-01

    Significant differences between the optical spectra taken from sound regions of teeth and carious regions have been observed. These differences appear both in absorption and in laser induced fluorescence spectra. Excitation by the 488 nm line of an argon ion laser beam showed a peak in the emission intensity around 553 nm for the sound dental material while the emission peak from the carious region was red-shifted by approximately 40 nm. The relative absorption of carious region was significantly higher at 488 nm; however its fluorescence intensity peak was lower by an order of magnitude compared to the sound tooth. Implications of these results for a safe, reliable and early detection of dental caries are discussed.

  10. Fluorescence and Spectral Imaging

    Directory of Open Access Journals (Sweden)

    Ralph S. DaCosta

    2007-01-01

    Full Text Available Early identification of dysplasia remains a critical goal for diagnostic endoscopy since early discovery directly improves patient survival because it allows endoscopic or surgical intervention with disease localized without lymph node involvement. Clinical studies have successfully used tissue autofluorescence with conventional white light endoscopy and biopsy for detecting adenomatous colonic polyps, differentiating benign hyperplastic from adenomas with acceptable sensitivity and specificity. In Barrett's esophagus, the detection of dysplasia remains problematic because of background inflammation, whereas in the squamous esophagus, autofluorescence imaging appears to be more dependable. Point fluorescence spectroscopy, although playing a crucial role in the pioneering mechanistic development of fluorescence endoscopic imaging, does not seem to have a current function in endoscopy because of its nontargeted sampling and suboptimal sensitivity and specificity. Other point spectroscopic modalities, such as Raman spectroscopy and elastic light scattering, continue to be evaluated in clinical studies, but still suffer the significant disadvantages of being random and nonimaging. A recent addition to the fluorescence endoscopic imaging arsenal is the use of confocal fluorescence endomicroscopy, which provides real-time optical biopsy for the first time. To improve detection of dysplasia in the gastrointestinal tract, a new and exciting development has been the use of exogenous fluorescence contrast probes that specifically target a variety of disease-related cellular biomarkers using conventional fluorescent dyes and novel potent fluorescent nanocrystals (i.e., quantum dots. This is an area of great promise, but still in its infancy, and preclinical studies are currently under way.

  11. Intrinsic fluorescence of protein in turbid media using empirical relation based on Monte Carlo lookup table

    Science.gov (United States)

    Einstein, Gnanatheepam; Udayakumar, Kanniyappan; Aruna, Prakasarao; Ganesan, Singaravelu

    2017-03-01

    Fluorescence of Protein has been widely used in diagnostic oncology for characterizing cellular metabolism. However, the intensity of fluorescence emission is affected due to the absorbers and scatterers in tissue, which may lead to error in estimating exact protein content in tissue. Extraction of intrinsic fluorescence from measured fluorescence has been achieved by different methods. Among them, Monte Carlo based method yields the highest accuracy for extracting intrinsic fluorescence. In this work, we have attempted to generate a lookup table for Monte Carlo simulation of fluorescence emission by protein. Furthermore, we fitted the generated lookup table using an empirical relation. The empirical relation between measured and intrinsic fluorescence is validated using tissue phantom experiments. The proposed relation can be used for estimating intrinsic fluorescence of protein for real-time diagnostic applications and thereby improving the clinical interpretation of fluorescence spectroscopic data.

  12. An approach to estimate spatial distribution of analyte within cells using spectrally-resolved fluorescence microscopy

    Science.gov (United States)

    Sharma, Dharmendar Kumar; Irfanullah, Mir; Basu, Santanu Kumar; Madhu, Sheri; De, Suman; Jadhav, Sameer; Ravikanth, Mangalampalli; Chowdhury, Arindam

    2017-03-01

    While fluorescence microscopy has become an essential tool amongst chemists and biologists for the detection of various analyte within cellular environments, non-uniform spatial distribution of sensors within cells often restricts extraction of reliable information on relative abundance of analytes in different subcellular regions. As an alternative to existing sensing methodologies such as ratiometric or FRET imaging, where relative proportion of analyte with respect to the sensor can be obtained within cells, we propose a methodology using spectrally-resolved fluorescence microscopy, via which both the relative abundance of sensor as well as their relative proportion with respect to the analyte can be simultaneously extracted for local subcellular regions. This method is exemplified using a BODIPY sensor, capable of detecting mercury ions within cellular environments, characterized by spectral blue-shift and concurrent enhancement of emission intensity. Spectral emission envelopes collected from sub-microscopic regions allowed us to compare the shift in transition energies as well as integrated emission intensities within various intracellular regions. Construction of a 2D scatter plot using spectral shifts and emission intensities, which depend on the relative amount of analyte with respect to sensor and the approximate local amounts of the probe, respectively, enabled qualitative extraction of relative abundance of analyte in various local regions within a single cell as well as amongst different cells. Although the comparisons remain semi-quantitative, this approach involving analysis of multiple spectral parameters opens up an alternative way to extract spatial distribution of analyte in heterogeneous systems. The proposed method would be especially relevant for fluorescent probes that undergo relatively nominal shift in transition energies compared to their emission bandwidths, which often restricts their usage for quantitative ratiometric imaging in

  13. Highly thermostable fluorescent proteins

    Science.gov (United States)

    Bradbury, Andrew M [Santa Fe, NM; Waldo, Geoffrey S [Santa Fe, NM; Kiss, Csaba [Los Alamos, NM

    2012-05-01

    Thermostable fluorescent proteins (TSFPs), methods for generating these and other stability-enhanced proteins, polynucleotides encoding such proteins, and assays and method for using the TSFPs and TSFP-encoding nucleic acid molecules are provided. The TSFPs of the invention show extremely enhanced levels of stability and thermotolerance. In one case, for example, a TSFP of the invention is so stable it can be heated to 99.degree. C. for short periods of time without denaturing, and retains 85% of its fluorescence when heated to 80.degree. C. for several minutes. The invention also provides a method for generating stability-enhanced variants of a protein, including but not limited to fluorescent proteins.

  14. Highly thermostable fluorescent proteins

    Science.gov (United States)

    Bradbury, Andrew M.; Waldo, Geoffrey S.; Kiss, Csaba

    2011-03-22

    Thermostable fluorescent proteins (TSFPs), methods for generating these and other stability-enhanced proteins, polynucleotides encoding such proteins, and assays and method for using the TSFPs and TSFP-encoding nucleic acid molecules are provided. The TSFPs of the invention show extremely enhanced levels of stability and thermotolerance. In one case, for example, a TSFP of the invention is so stable it can be heated to 99.degree. C. for short periods of time without denaturing, and retains 85% of its fluorescence when heated to 80.degree. C. for several minutes. The invention also provides a method for generating stability-enhanced variants of a protein, including but not limited to fluorescent proteins.

  15. Origin of tryptophan fluorescence lifetimes. Part 2: fluorescence lifetimes origin of tryptophan in proteins.

    Science.gov (United States)

    Albani, J R

    2014-01-01

    Fluorescence intensity decays of L-tryptophan in proteins dissolved in pH 7 buffer, in ethanol and in 6 M guanidine pH 7.8 and in lyophilized proteins were measured. In all protein conditions, three lifetimes were obtained along the emission spectrum (310-410 nm). The two shortest lifetimes are in the same range of those obtained for L-Trp in water or in ethanol. Thus, these two lifetimes originate from specific two sub-structures existing in the excited state and are inherent to the tryptophan structure independently of the surrounding environment (amino acids residues, solvent, etc.) In proteins, the third lifetime originates from the interactions that are occurring between tryptophan residues and neighboring amino acids. Populations of these lifetimes are independent of the excitation wavelength and thus originate from pre-defined sub structures existing in the excited state and put into evidence after tryptophan excitation. Fluorescence decay studies of different tripeptides having a tryptophan residue in second position show that the best analysis is obtained with two fluorescence lifetimes. Consequently, this result seems to exclude the possibility that peptide bond induces the third fluorescence lifetimes. Indole dissolved in water and/or in ethanol emits with two fluorescence lifetimes that are completely different from those observed for L-Trp. Absence of the third lifetime in ethanol demonstrates that indole behaves differently when compared to tryptophan. Thus, it seems not adequate to attribute fluorescence lifetime or fluorescence properties of tryptophan to indole ring and to compare tryptophan fluorescence properties in proteins to molecules having close structures such as NATA which fluoresces with one lifetime.

  16. Quantitative X-ray fluorescence analysis of samples of less than 'infinite thickness': Difficulties and possibilities

    Energy Technology Data Exchange (ETDEWEB)

    Sitko, Rafal, E-mail: rafal.sitko@us.edu.p [Institute of Chemistry, Silesian University, 40-006 Katowice (Poland)

    2009-11-15

    X-ray fluorescence spectrometry due to its nondestructive nature is widely applied in analysis of single layers and multiple layer films (e.g. semiconductors, electrooptic and solar cell devices, coatings, corrosion and paint layers), individual particles (airborne, fly ash, gunshot residue particles, etc.), art and archeological objects (manuscripts, paintings, icons) and many others. Quantitative analysis of these materials, frequently classified as samples of less than infinite thickness (thin or intermediate-thickness samples), required applying adequate matrix correction methods taking into account complex dependence of analyte fluorescent radiation intensity on full matrix composition and sample thickness. In this article, the matrix correction methods including fundamental parameters, Monte Carlo simulations, influence coefficients algorithms and methods based on X-ray transmission measurements are reviewed. The difficulties in the analysis of single layer and multiple layer films and the accuracy of fundamental parameter methods in simultaneous determination of their thickness and composition are discussed. The quantitative analysis of individual particles and inhomogeneous and/or complex structure materials using fundamental parameter and Monte Carlo simulation methods in micro-beam X-ray fluorescence spectrometry are also reviewed. Some references are devoted to the analysis of light matrix samples, e.g. geological, environmental and biological samples, in which undetectable low-Z elements are present (so-called 'dark matrix') using backscattered fundamental parameter methods. Since the samples of less than infinite thickness are partially transparent for X-ray beams, the transmission measurements present possibilities that are unattainable for bulk samples. Thus, the emission-transmission method and also new instruments allowing measurements of the primary X-ray beam transmitted through the sample together with measurements of X

  17. Multispectral fluorescence imaging techniques for nondestructive food safety inspection

    Science.gov (United States)

    Kim, Moon S.; Lefcourt, Alan M.; Chen, Yud-Ren

    2004-03-01

    The use of spectral sensing has gained acceptance as a rapid means for nondestructive inspection of postharvest food produce. Current technologies generally use color or a single wavelength camera technology. The applicability and sensitivity of these techniques can be expanded through the use of multiple wavelengths. Reflectance in the Vis/NIR is the prevalent spectral technique. Fluorescence, compared to reflectance, is regarded as a more sensitive technique due to its dynamic responses to subtle changes in biological entities. Our laboratory has been exploring fluorescence as a potential means for detection of quality and wholesomeness of food products. Applications of fluorescence sensing require an understanding of the spectral characteristics emanating from constituents and potential contaminants. A number of factors affecting fluorescence emission characteristics are discussed. Because of relatively low fluorescence quantum yield from biological samples, a system with a powerful pulse light source such as a laser coupled with a gated detection device is used to harvest fluorescence, in the presence of ambient light. Several fluorescence sensor platforms developed in our laboratory, including hyperspectral imaging, and laser-induced fluorescence (LIF) and steady-state fluorescence imaging systems with multispectral capabilities are presented. We demonstrate the potential uses of recently developed fluorescence imaging platforms in food safety inspection of apples contaminated with animal feces.

  18. Fluorescent Nanoparticle Uptake for Brain Tumor Visualization

    Directory of Open Access Journals (Sweden)

    Rachel Tréhin

    2006-04-01

    Full Text Available Accurate delineation of tumor margins is vital to the successful surgical resection of brain tumors. We have previously developed a multimodal nanoparticle CLIO-Cy5.5, which is detectable by both magnetic resonance imaging and fluorescence, to assist in intraoperatively visualizing tumor boundaries. Here we examined the accuracy of tumor margin determination of orthotopic tumors implanted in hosts with differing immune responses to the tumor. Using a nonuser-based signal intensity method applied to fluorescent micrographs of 9L gliosarcoma green fluorescent protein (GFP tumors, mean overestimations of 2 and 24 µm were obtained using Cy5.5 fluorescence, compared to the true tumor margin determined by GFP fluorescence, in nude mice and rats, respectively. To resolve which cells internalized the nanoparticle and to quantitate degree of uptake, tumors were disaggregated and cells were analyzed by flow cytometry and fluorescence microscopy. Nanoparticle uptake was seen in both CD11b+ cells (representing activated microglia and macrophages and tumor cells in both animal models by both methods. CD11b+ cells were predominantly found at the tumor margin in both hosts, but were more pronounced at the margin in the rat model. Additional metastatic (CT26 colon and primary (Gli36 glioma brain tumor models likewise demonstrated that the nanoparticle was internalized both by tumor cells and by host cells. Together, these observations suggest that fluorescent nanoparticles provide an accurate method of tumor margin estimation based on a combination of tumor cell and host cell uptake for primary and metastatic tumors in animal model systems and offer potential for clinical translation.

  19. Recent Progress on Plasmon-Enhanced Fluorescence

    Directory of Open Access Journals (Sweden)

    Dong Jun

    2015-12-01

    Full Text Available The optically generated collective electron density waves on metal–dielectric boundaries known as surface plasmons have been of great scientific interest since their discovery. Being electromagnetic waves on gold or silver nanoparticle’s surface, localised surface plasmons (LSP can strongly enhance the electromagnetic field. These strong electromagnetic fields near the metal surfaces have been used in various applications like surface enhanced spectroscopy (SES, plasmonic lithography, plasmonic trapping of particles, and plasmonic catalysis. Resonant coupling of LSPs to fluorophore can strongly enhance the emission intensity, the angular distribution, and the polarisation of the emitted radiation and even the speed of radiative decay, which is so-called plasmon enhanced fluorescence (PEF. As a result, more and more reports on surface-enhanced fluorescence have appeared, such as SPASER-s, plasmon assisted lasing, single molecule fluorescence measurements, surface plasmoncoupled emission (SPCE in biological sensing, optical orbit designs etc. In this review, we focus on recent advanced reports on plasmon-enhanced fluorescence (PEF. First, the mechanism of PEF and early results of enhanced fluorescence observed by metal nanostructure will be introduced. Then, the enhanced substrates, including periodical and nonperiodical nanostructure, will be discussed and the most important factor of the spacer between molecule and surface and wavelength dependence on PEF is demonstrated. Finally, the recent progress of tipenhanced fluorescence and PEF from the rare-earth doped up-conversion (UC and down-conversion (DC nanoparticles (NPs are also commented upon. This review provides an introduction to fundamentals of PEF, illustrates the current progress in the design of metallic nanostructures for efficient fluorescence signal amplification that utilises propagating and localised surface plasmons.

  20. Recent Progress on Plasmon-Enhanced Fluorescence

    Science.gov (United States)

    Dong, Jun; Zhang, Zhenglong; Zheng, Hairong; Sun, Mentao

    2015-12-01

    The optically generated collective electron density waves on metal-dielectric boundaries known as surface plasmons have been of great scientific interest since their discovery. Being electromagnetic waves on gold or silver nanoparticle's surface, localised surface plasmons (LSP) can strongly enhance the electromagnetic field. These strong electromagnetic fields near the metal surfaces have been used in various applications like surface enhanced spectroscopy (SES), plasmonic lithography, plasmonic trapping of particles, and plasmonic catalysis. Resonant coupling of LSPs to fluorophore can strongly enhance the emission intensity, the angular distribution, and the polarisation of the emitted radiation and even the speed of radiative decay, which is so-called plasmon enhanced fluorescence (PEF). As a result, more and more reports on surface-enhanced fluorescence have appeared, such as SPASER-s, plasmon assisted lasing, single molecule fluorescence measurements, surface plasmoncoupled emission (SPCE) in biological sensing, optical orbit designs etc. In this review, we focus on recent advanced reports on plasmon-enhanced fluorescence (PEF). First, the mechanism of PEF and early results of enhanced fluorescence observed by metal nanostructure will be introduced. Then, the enhanced substrates, including periodical and nonperiodical nanostructure, will be discussed and the most important factor of the spacer between molecule and surface and wavelength dependence on PEF is demonstrated. Finally, the recent progress of tipenhanced fluorescence and PEF from the rare-earth doped up-conversion (UC) and down-conversion (DC) nanoparticles (NPs) are also commented upon. This review provides an introduction to fundamentals of PEF, illustrates the current progress in the design of metallic nanostructures for efficient fluorescence signal amplification that utilises propagating and localised surface plasmons.

  1. Fluorescein Derivatives in Intravital Fluorescence Imaging

    Directory of Open Access Journals (Sweden)

    Michael S. Roberts

    2013-08-01

    Full Text Available Intravital fluorescence microscopy enables the direct imaging of fluorophores in vivo and advanced techniques such as fluorescence lifetime imaging (FLIM enable the simultaneous detection of multiple fluorophores. Consequently, it is now possible to record distribution and metabolism of a chemical in vivo and to optimise the delivery of fluorophores in vivo. Recent clinical applications with fluorescein and other intravital fluorescent stains have occurred in neurosurgery, dermatology [including photodynamic therapy (PDT] and endomicroscopy. Potential uses have been identified in periodontal disease, skin graft and cancer surgery. Animal studies have demonstrated that diseased tissue can be specifically stained with fluorophore conjugates. This review focuses on the fluorescein derived fluorophores in common clinical use and provides examples of novel applications from studies in tissue samples.

  2. Introduction to fluorescence

    CERN Document Server

    Jameson, David M

    2014-01-01

    "An essential contribution to educating scientists in the principles of fluorescence. It will also be an important addition to the libraries of practitioners applying the principles of molecular fluorescence."-Ken Jacobson, Kenan Distinguished Professor of Cell Biology and Physiology, University of North Carolina at Chapel Hill"An exquisite compendium of fluorescence and its applications in biochemistry enriched by a very exciting historical perspective. This book will become a standard text for graduate students and other scientists."-Drs. Zygmunt (Karol) Gryczynski and Ignacy Gryczynski, University of North Texas Health Science Center"… truly a masterwork, combining clarity, precision, and good humor. The reader, novice or expert, will be pleased with the text and will not stop reading. It is a formidable account of the fluorescence field, which has impacted the life sciences so considerably in the last 60 years."-Jerson L. Silva, M.D., Ph.D., Professor and Director, National Institute of Science and Tech...

  3. Fluorescence optimisation and lifetime studies of fingerprints treated with magnetic powders.

    Science.gov (United States)

    Seah, L K; Dinish, U S; Phang, W F; Chao, Z X; Murukeshan, V M

    2005-09-10

    Fluorescence study plays a significant role in fingerprint detection when conventional chemical enhancement methods fail. The basic properties of fluorescence emission such as colour, intensity and lifetime could be well exploited in the detection of latent fingerprints under steady state and in dynamic methods. This paper describes a systematic study of fluorescence emission intensity from fingerprint samples treated with different magnetic powders. Understanding of suitable excitation wavelength required for getting maximum fluorescence emission intensity could be beneficial when selecting the appropriate fluorescent powders for the fingerprint detection. Lifetime study of fingerprints treated with various magnetic powders was also carried out. The importance of lifetime study is well explained through the time-resolved (TR) imaging of fingerprints with nanosecond resolution. Results from the TR imaging study revealed an improvement in the fingerprint image contrast. This is significant when the print is deposited on fluorescing background and its emission wavelength is close to that of treated fingerprint.

  4. Functional Fluorescent Organic Nanoparticles

    OpenAIRE

    Campioli, Elisa

    2013-01-01

    This thesis presents an extensive study on fluorescent organic nanoparticles and fluorescent organic binary and ternary nanoassemblies. In particular the attention is focused on the preparation and characterization of organic nanoparticles and new nanocomposites obtained from different types of small organic molecules, their stabilization and the use of these materials for biological and optoelectronics applications. The work deals at the beginning with the description of some methods used...

  5. Optical modulator based on propagating surface plasmon coupled fluorescent thin film: proof-of-concept studies

    Science.gov (United States)

    Cao, Shuo-Hui; Wang, Zheng-Chuang; Weng, Yu-Hua; Xie, Kai-Xin; Chen, Min; Zhai, Yan-Yun; Li, Yao-Qun

    2017-06-01

    We demonstrate that the propagating surface plasmon coupled fluorescent thin film can be utilized as a fluorescence modulator to mimic multiple representative Boolean logic operations. Surface plasmon mediated fluorescence presents characteristic properties including directional and polarized emission, which hold the feasibility in creating a universal optical modulator. In this work, through constructing the thin layer with the specific thickness, surface plasmon mediated fluorescence can be modulated with an ON-OFF ratio by more than 5-fold, under a series of coupling configurations.

  6. Anthracene Fluorescence Quenching by a Tetrakis (Ketocarboxamide Cavitand

    Directory of Open Access Journals (Sweden)

    Tibor Zoltan Janosi

    2014-01-01

    Full Text Available Quenching of both fluorescence lifetime and fluorescence intensity of anthracene was investigated in the presence of a newly derived tetrakis (ketocarboxamide cavitand at various concentrations. Time-correlated single photon counting method was applied for the lifetime measurements. A clear correlation between the fluorescence lifetime of anthracene as a function of cavitand concentration in dimethylformamide solution was observed. The bimolecular collisional quenching constant was derived from the decrease of lifetime. Fluorescence intensity was measured in the emission wavelength region around 400 nm as a result of excitation at 280 nm. Effective quenching was observed in the presence of the cavitand. The obtained Stern-Volmer plot displayed upward curvature. The results did not follow even extended Stern-Volmer behavior, often used to describe deviations from static bimolecular quenching. To explain our results we adopted the Smoluchowski model and obtained a reasonable estimate for the molecular radius of the cavitand in solution.

  7. Sequence-Dependent Fluorescence of Cy3- and Cy5-Labeled Double-Stranded DNA.

    Science.gov (United States)

    Kretschy, Nicole; Sack, Matej; Somoza, Mark M

    2016-03-16

    The fluorescent intensity of Cy3 and Cy5 dyes is strongly dependent on the nucleobase sequence of the labeled oligonucleotides. Sequence-dependent fluorescence may significantly influence the data obtained from many common experimental methods based on fluorescence detection of nucleic acids, such as sequencing, PCR, FRET, and FISH. To quantify sequence dependent fluorescence, we have measured the fluorescence intensity of Cy3 and Cy5 bound to the 5' end of all 1024 possible double-stranded DNA 5mers. The fluorescence intensity was also determined for these dyes bound to the 5' end of fixed-sequence double-stranded DNA with a variable sequence 3' overhang adjacent to the dye. The labeled DNA oligonucleotides were made using light-directed, in situ microarray synthesis. The results indicate that the fluorescence intensity of both dyes is sensitive to all five bases or base pairs, that the sequence dependence is stronger for double- (vs single-) stranded DNA, and that the dyes are sensitive to both the adjacent dsDNA sequence and the 3'-ssDNA overhang. Purine-rich sequences result in higher fluorescence. The results can be used to estimate measurement error in experiments with fluorescent-labeled DNA, as well as to optimize the fluorescent signal by considering the nucleobase environment of the labeling cyanine dye.

  8. Real-time quantitative fluorescence measurement of microscale cell culture analog systems

    Science.gov (United States)

    Oh, Taek-il; Kim, Donghyun; Tatosian, Daniel; Sung, Jong Hwan; Shuler, Michael

    2007-02-01

    A microscale cell culture analog (μCCA) is a cell-based lab-on-a-chip assay that, as an animal surrogate, is applied to pharmacological studies for toxicology tests. A μCCA typically comprises multiple chambers and microfluidics that connect the chambers, which represent animal organs and blood flow to mimic animal metabolism more realistically. A μCCA is expected to provide a tool for high-throughput drug discovery. Previously, a portable fluorescence detection system was investigated for a single μCCA device in real-time. In this study, we present a fluorescence-based imaging system that provides quantitative real-time data of the metabolic interactions in μCCAs with an emphasis on measuring multiple μCCA samples simultaneously for high-throughput screening. The detection system is based on discrete optics components, with a high-power LED and a charge-coupled device (CCD) camera as a light source and a detector, for monitoring cellular status on the chambers of each μCCA sample. Multiple samples are characterized mechanically on a motorized linear stage, which is fully-automated. Each μCCA sample has four chambers, where cell lines MES-SA/DX- 5, and MES-SA (tumor cells of human uterus) have been cultured. All cell-lines have been transfected to express the fusion protein H2B-GFP, which is a human histone protein fused at the amino terminus to EGFP. As a model cytotoxic drug, 10 μM doxorubicin (DOX) was used. Real-time quantitative data of the intensity loss of enhanced green fluorescent protein (EGFP) during cell death of target cells have been collected over several minutes to 40 hours. Design issues and improvements are also discussed.

  9. Combined influences of chromatic aberration and scattering in depth-resolved two-photon fluorescence endospectroscopy.

    Science.gov (United States)

    Wu, Yicong; Li, Xingde

    2010-10-27

    The influence of chromatic aberration of an objective lens in two-photon fluorescence (TPF) endospectroscopy of scattering media has been systematically investigated through both experiments and numerical simulations. Experiments were carried out on a uniform 3D scattering gelatin phantom embedded with TiO(2) granules (to mimic tissue scattering) and fluorescein-tagged polystyrene beads. It was found that fluorescence spectral intensity and lineshape varied as a function of depth when measured with a gradient-index (GRIN) lens which has severe chromatic aberration. The spectral distortion caused by the chromatic aberration became diminishing as the imaging depth increased. Ray tracing analysis and Monte Carlo simulations were carried out to study the interplay of chromatic aberration and scattering in the depth-resolved TPF spectra. The simulation results suggest that the collected fluorescence signals from deeper layers included more out-of-focus photons that experienced a few or multiple scatterings, which diminish the influence of chromatic aberration on the measured TPF spectra. The simulated collection efficiencies of TPF at different wavelengths and depths can be used to properly recover the true depth-resolved TPF spectra of a relatively uniform scattering medium.

  10. Deconvolution of calcium fluorescent indicator signal from AFM cantilever reflection.

    Science.gov (United States)

    Lopez-Ayon, G Monserratt; Oliver, David J; Grutter, Peter H; Komarova, Svetlana V

    2012-08-01

    Atomic force microscopy (AFM) can be combined with fluorescence microscopy to measure the changes in intracellular calcium levels (indicated by fluorescence of Ca²⁺ sensitive dye fluo-4) in response to mechanical stimulation performed by AFM. Mechanical stimulation using AFM is associated with cantilever movement, which may interfere with the fluorescence signal. The motion of the AFM cantilever with respect to the sample resulted in changes of the reflection of light back to the sample and a subsequent variation in the fluorescence intensity, which was not related to changes in intracellular Ca²⁺ levels. When global Ca²⁺ responses to a single stimulation were assessed, the interference of reflected light with the fluorescent signal was minimal. However, in experiments where local repetitive stimulations were performed, reflection artifacts, correlated with cantilever motion, represented a significant component of the fluorescent signal. We developed a protocol to correct the fluorescence traces for reflection artifacts, as well as photobleaching. An added benefit of our method is that the cantilever reflection in the fluorescence recordings can be used for precise temporal correlation of the AFM and fluorescence measurements.

  11. Polyester Fabric's Fluorescent Dyeing in Supercritical Carbon Dioxide and its Fluorescence Imaging.

    Science.gov (United States)

    Xiong, Xiaoqing; Xu, Yanyan; Zheng, Laijiu; Yan, Jun; Zhao, Hongjuan; Zhang, Juan; Sun, Yanfeng

    2017-03-01

    As one of the most important coumarin-like dyes, disperse fluorescent Yellow 82 exhibits exceptionally large two-photon effects. Here, it was firstly introduced into the supercritical CO2 dyeing polyester fabrics in this work. Results of the present work showed that the dyeing parameters such as the dyeing time, pressure and temperature had remarkable influences on the color strength of fabrics. The optimized dyeing condition in supercritical CO2 dyeing has been proposed that the dyeing time was 60 min; the pressure was 25 MPa and the temperature was 120 °C. As a result, acceptable products were obtained with the wash and rub fastness rating at 5 or 4-5. The polyester fabrics dyed with fluorescent dyes can be satisfied for the requirement of manufacturing warning clothing. Importantly, the confocal microscopy imaging technology was successfully introduced into textile fields to observe the distribution and fluorescence intensity of disperse fluorescent Yellow 82 on polyester fabrics. As far as we know, this is the first report about supercritical CO2 dyeing polyester fabrics based on disperse fluorescent dyes. It will be very helpful for the further design of new fluorescent functional dyes suitable for supercritical CO2 dyeing technique.

  12. Photonic Crystal Enhanced Fluorescence for Early Breast Cancer Biomarker Detection

    OpenAIRE

    Cunningham, Brian T.; Zangar, Richard C.

    2012-01-01

    Photonic crystal surfaces offer a compelling platform for improving the sensitivity of surface-based fluorescent assays used in disease diagnostics. Through the complementary processes of photonic crystal enhanced excitation and enhanced extraction, a periodic dielectric-based nanostructured surface can simultaneously increase the electric field intensity experienced by surface-bound fluorophores and increase the collection efficiency of emitted fluorescent photons. Through the ability to ine...

  13. Light emission from compound eye with conformal fluorescent coating

    Science.gov (United States)

    Martín-Palma, Raúl J.; Miller, Amy E.; Pulsifer, Drew P.; Lakhtakia, Akhlesh

    2015-03-01

    Compound eyes of insects are attractive biological systems for engineered biomimicry as artificial sources of light, given their characteristic wide angular field of view. A blowfly eye was coated with a thin conformal fluorescent film, with the aim of achieving wide field-of-view emission. Experimental results showed that the coated eye emitted visible light and that the intensity showed a weaker angular dependence than a fluorescent thin film deposited on a flat surface.

  14. Boron Polylactide Nanoparticles Exhibiting Fluorescence and Phosphorescence in Aqueous Medium

    Science.gov (United States)

    Pfister, Anne; Zhang, Guoqing; Zareno, Jessica; Horwitz, Alan F.; Fraser, Cassandra L.

    2008-01-01

    Difluoroboron dibenzoylmethane-polylactide, BF2dbmPLA, a biocompatible polymerluminophore conjugate was fabricated as nanoparticles. Spherical particles <100 nm in size were generated via nanoprecipitation. Intense blue fluorescence, two-photon absorption, and long-lived room temperature phosphorescence (RTP) are retained in aqueous suspension. The nanoparticles were internalized by cells and visualized by fluorescence microscopy. Luminescent boron biomaterials show potential for imaging and sensing. PMID:19081748

  15. A cluster randomised trial, cost-effectiveness analysis and psychosocial evaluation of insulin pump therapy compared with multiple injections during flexible intensive insulin therapy for type 1 diabetes: the REPOSE Trial.

    Science.gov (United States)

    Heller, Simon; White, David; Lee, Ellen; Lawton, Julia; Pollard, Daniel; Waugh, Norman; Amiel, Stephanie; Barnard, Katharine; Beckwith, Anita; Brennan, Alan; Campbell, Michael; Cooper, Cindy; Dimairo, Munyaradzi; Dixon, Simon; Elliott, Jackie; Evans, Mark; Green, Fiona; Hackney, Gemma; Hammond, Peter; Hallowell, Nina; Jaap, Alan; Kennon, Brian; Kirkham, Jackie; Lindsay, Robert; Mansell, Peter; Papaioannou, Diana; Rankin, David; Royle, Pamela; Smithson, W Henry; Taylor, Carolin

    2017-04-01

    Insulin is generally administered to people with type 1 diabetes mellitus (T1DM) using multiple daily injections (MDIs), but can also be delivered using infusion pumps. In the UK, pumps are recommended for patients with the greatest need and adult use is less than in comparable countries. Previous trials have been small, of short duration and have failed to control for training in insulin adjustment. To assess the clinical effectiveness and cost-effectiveness of pump therapy compared with MDI for adults with T1DM, with both groups receiving equivalent structured training in flexible insulin therapy. Pragmatic, multicentre, open-label, parallel-group cluster randomised controlled trial, including economic and psychosocial evaluations. After participants were assigned a group training course, courses were randomly allocated in pairs to either pump or MDI. Eight secondary care diabetes centres in the UK. Adults with T1DM for > 12 months, willing to undertake intensive insulin therapy, with no preference for pump or MDI, or a clinical indication for pumps. Pump or MDI structured training in flexible insulin therapy, followed up for 2 years. MDI participants used insulin analogues. Pump participants used a Medtronic Paradigm(®) Veo(TM) (Medtronic, Watford, UK) with insulin aspart (NovoRapid, Novo Nordisk, Gatwick, UK). Primary outcome - change in glycated haemoglobin (HbA1c) at 2 years in participants whose baseline HbA1c was ≥ 7.5% (58 mmol/mol). Key secondary outcome - proportion of participants with HbA1c ≤ 7.5% at 2 years. Other outcomes at 6, 12 and 24 months - moderate and severe hypoglycaemia; insulin dose; body weight; proteinuria; diabetic ketoacidosis; quality of life (QoL); fear of hypoglycaemia; treatment satisfaction; emotional well-being; qualitative interviews with participants and staff (2 weeks), and participants (6 months); and ICERs in trial and modelled estimates of cost-effectiveness. We randomised 46 courses comprising 317 participants

  16. Lifetime Resolved Fluorescence Fluctuation Spectroscopy

    Science.gov (United States)

    Guo, Peng; Berland, Keith

    2009-11-01

    Fluorescence correlation spectroscopy (FCS) has been widely used investigate molecular dynamics and interactions in biological systems. FCS typically resolves the component species of a sample either through differences in diffusion coefficient or molecular brightness. Diffusion based assays currently have a major limitation which requires that the diffusion coefficients of component species in a sample must be substantially different in order to be resolved. This criterion is not met in many important cases, such as when molecules of similar molecular weight bind to each other. This limitation can be overcome, and resolution of FCS measurements enhanced, by combining FCS measurements with measurements of fluorescence lifetimes. By using of global analysis on simultaneously acquired FCS and lifetime data we show that we can dramatically enhance resolution in FCS measurements, and accurately resolve the concentration and diffusion coefficients of multiple sample components even when their diffusion coefficients are identical provided there is a difference in the lifetime of the component species. We show examples of this technique using both simulations and experiments. It is expected that this method will be of significance for binding assays studying molecular interactions.

  17. Near-membrane refractometry using supercritical angle fluorescence

    CERN Document Server

    Brunstein, Maia; Oheim, Martin

    2016-01-01

    Total internal reflection fluorescence (TIRF) microscopy and its variants are key technologies for visualizing the dynamics of single molecules or organelles in live cells. Yet, truely quantitative TIRF remains problematic. One unknown hampering the interpretation of evanescent-wave excited fluorescence intensities is the undetermined cell refractive index (RI). Here, we use a combination of TIRF excitation and supercritical angle fluorescence emission detection to directly mea-sure the average RI in the 'footprint' region of the cell, during imaging. Our RI measurement is based on the determination on a back-focal plane image of the critical angle separating supercritical and undercritial fluorescence emission components. We validate our method by imaging mouse embryonic fibroblasts. By targeting various dyes and fluorescent-protein chimerae to vesicles, the plasma membrane as well as mitochondria and the ER, we demonstrate local RI measurements with subcellular resolution on a standard TIRF microscope with ...

  18. Fluorescent angiography of chicken embryo and photobleaching velocimetry

    Science.gov (United States)

    Namykin, Anton A.; Stiukhina, Elena S.; Fedosov, Ivan V.; Postnov, Dmitry E.; Tuchin, Valery V.

    2017-03-01

    Fluorescent angiography approach in application to a living chicken embryo is discussed. It provides precise vessel wall detection and demonstrates usefulness for real time monitoring of vasoconstriction and vasodilatation related to self regulation of vascular network as well as to response to external factors. On the other hand, high stability of fluorescence and long period of dye elimination makes variations of fluorescent intensity practically independent from fast variations of blood flow rate. Therefore, we proposed the improvement of fluorescent angiography technique by introduction of photobleaching fluorescent velocimetry approach. We have developed the imaging system for intravital microscopic photobleaching velocimetry and tested it by using a glass capillary tube as a model of blood vessel. We demonstrated high potential of the technique for instant flow velocity distribution profile measurement with high spatial and temporal resolution up to 2 μm and 60 ms, respectively.

  19. Octyl gallate: An antioxidant demonstrating selective and sensitive fluorescent property.

    Science.gov (United States)

    Wang, Qing; Zhang, Yongkui; Li, Hui

    2017-03-15

    Octyl gallate (OG) is an internationally recognized antioxidant that demonstrates selective and sensitive fluorescent property. The fluorescence of OG can be selectively enhanced in the presence of human serum albumin (HSA) and bovine serum albumin (BSA). The specific structures of HSA and BSA provided the basic conditions for fluorescence enhancement. OG yielded approximately 49- and 11-fold increments in emission intensity in the presence of HSA and BSA at a molar ratio of 1:1, respectively. The lifetimes of HSA and BSA correspondingly decreased. A Förster resonance energy transfer phenomenon occurred during interaction between OG and HSA or BSA. Our in-depth investigation of OG-HSA interaction showed that formation of a stable complex was an important prerequisite to efficiently enhance the fluorescence of OG. The selective and sensitive fluorescent property of OG can possibly be used to determine OG concentration via the standard addition method, which must be performed under certain conditions.

  20. Quenching of chlorophyll fluorescence induced by silver nanoparticles

    Science.gov (United States)

    Queiroz, A. M.; Mezacasa, A. V.; Graciano, D. E.; Falco, W. F.; M'Peko, J.-C.; Guimarães, F. E. G.; Lawson, T.; Colbeck, I.; Oliveira, S. L.; Caires, A. R. L.

    2016-11-01

    The interaction between chlorophyll (Chl) and silver nanoparticles (AgNPs) was evaluated by analyzing the optical behavior of Chl molecules surrounded by different concentrations of AgNPs (10, 60, and 100 nm of diameter). UV-Vis absorption, steady state and time-resolved fluorescence measurements were performed for Chl in the presence and absence of these nanoparticles. AgNPs strongly suppressed the Chl fluorescence intensity at 678 nm. The Stern-Volmer constant (KSV) showed that fluorescence suppression is driven by the dynamic quenching process. In particular, KSV was nanoparticle size-dependent with an exponential decrease as a function of the nanoparticle diameter. Finally, changes in the Chl fluorescence lifetime in the presence of nanoparticles demonstrated that the fluorescence quenching may be induced by the excited electron transfer from the Chl molecules to the metal nanoparticles.

  1. Fluorescence dynamics of human epidermis (ex vivo) and skin (in vivo)

    Science.gov (United States)

    Salomatina, Elena V.; Pravdin, Alexander B.

    2003-10-01

    The temporal behavior of autofluorescence of human skin and epidermis under continuous UV-irradiation has been studied. Fluorescence spectra and kinetic curves of fluorescence intensity have been obtained. The fluorescence intensity recovery after dark period also has been examined. The vitiligo skin and epidermis were used for comparing their spectra with reflectance and fluorescence spectra of healthy skin. The epidermal samples were prepared using surface epidermis stripping technique. It has been concluded that fluorophores being undergone the UVA photobleaching are actually present in epidermal layer, and immediate pigment darkening does contribute, no less than a half of magnitude, to the autofluorescence decrease under continuous UVA irradiation.

  2. Finding of Optimal Calcium Ion Probes for Fluorescence Lifetime Measurement

    Science.gov (United States)

    Yoshiki, Keisuke; Azuma, Hiroki; Yoshioka, Kazuhiko; Hashimoto, Mamoru; Araki, Tsutomu

    We have investigated the fluorescence lifetime properties of 8 calcium ion probes, calcium-green-1, calcium green-2, calcium green-5N, calcium orange, oregon green 488 BAPTA-6F, fluo-3, fluo-4, and fluo-5N. We found that the decay time of calcium green-5N varied more sensitively with calcium concentration than calcium green-1 which was known to be a highly sensitive probe. We have also found that the center of observable range of calcium concentration by fluorescence lifetime measurement is lower than that by fluorescence intensity measurement.

  3. The nature of multiphoton fluorescence from red blood cells

    Science.gov (United States)

    Saytashev, Ilyas; Murphy, Michael; Osseiran, Sam; Spence, Dana M.; Evans, Conor L.; Dantus, Marcos

    2016-03-01

    We report on the nature of multiphoton excited fluorescence observed from human erythrocytes (red blood cells RBC's) and their "ghosts" following 800nm sub-15 fs excitation. The detected optical signal is assigned as two-photon excited fluorescence from hemoglobin. Our findings are supported by wavelength-resolved fluorescence lifetime decay measurements using time-correlated single photon counting system from RBC's, their ghosts as well as in vitro samples of various fluorophores including riboflavin, NADH, NAD(P)H, hemoglobin. We find that low-energy and short-duration pulses allow two-photon imaging of RBC's, but longer more intense pulses lead to their destruction.

  4. Analyzing of LED-induced Blood Fluorescent Spectra

    Institute of Scientific and Technical Information of China (English)

    高淑梅; 骆晓森; 兰秀风; 刘莹; 陆健; 倪晓武

    2002-01-01

    The visible fluorescence spectra of healthy mice blood cells have been measured by light emitting diode (LED) excitation at about 570 nm (Δλ1/2≈32 nm ) in vitro. It is found that the spectral profiles of mouse blood, erythrocyte and hemoglobin are similar. The physical mechanism for LED-induced blood cells fluorescence spectra is analyzed. The development of low intensity laser or light irradiating blood therapy in vivo and in vitro may be guided by the understanding of blood fluorescence characteristics.

  5. Preparation and Characterization of Fluorescence Probe from Assembly Hydroxyapatite Nanocomposite

    Directory of Open Access Journals (Sweden)

    Li Guang-Ming

    2010-01-01

    Full Text Available Abstract A new nanocomposite fluorescence probe with thioglycolic acid (TA functional layers embedded inside the hydroxyapatite nanoribbon spherulites has been synthesized. The fluorescence intensity of the novel probe is about 1.5–3.3-fold increase compared with the probe containing no TA. When used to detect cadmium ion, the most of original assembly nanoribbon spherulites structure in the novel probe is found to have been damaged to new flake structures. The mechanism of determining cadmium ion in alcohol solution has been studied. The present systematic study provides significant information on the effect of assembly nanostructure on the metal-enhanced fluorescence phenomenon.

  6. Genetic Analysis of Digestive Physiology Using Fluorescent Phospholipid Reporters

    Science.gov (United States)

    Farber, Steven A.; Pack, Michael; Ho, Shiu-Ying; Johnson, Iain D.; Wagner, Daniel S.; Dosch, Roland; Mullins, Mary C.; Hendrickson, H. Stewart; Hendrickson, Elizabeth K.; Halpern, Marnie E.

    2001-05-01

    Zebrafish are a valuable model for mammalian lipid metabolism; larvae process lipids similarly through the intestine and hepatobiliary system and respond to drugs that block cholesterol synthesis in humans. After ingestion of fluorescently quenched phospholipids, endogenous lipase activity and rapid transport of cleavage products results in intense gall bladder fluorescence. Genetic screening identifies zebrafish mutants, such as fat free, that show normal digestive organ morphology but severely reduced phospholipid and cholesterol processing. Thus, fluorescent lipids provide a sensitive readout of lipid metabolism and are a powerful tool for identifying genes that mediate vertebrate digestive physiology.

  7. Research of the fluorescence detection apparatus for nutrients

    Science.gov (United States)

    Wang, Yu; Yan, Huimin; Ni, Xuxiang; Xu, Xiaoyi; Chen, Shibing

    2015-10-01

    The research of the multifunctional analyzer of Clinical Nutrition, which integrates the absorbance, luminescence, fluorescence and other optical detection methods, can overcome the functional limitations of a single technology on human nutrition analysis, and realize a rapid and accurate analysis of the nutrients. This article focuses on the design of fluorescence detection module that uses a photomultiplier tube(PMT) to detect weak fluorescence, and utilizes the single photon counting method to measure the fluorescence intensity, and then according to the relationship between the fluorescent marker and fluorescence intensity, the concentration of the analyte can be derived. Using fluorescein isothiocyanate(FITC, the most widely used fluorescein currently)to mark antibodies in the experiment, therefore, according to the maximum absorption wavelength and the maximum emission wavelength of the fluorescein isothiocyanate, to select the appropriate filters to set up the optical path. In addition, the fluorescence detection apparatus proposed in this paper uses an aspherical lens with large numerical aperture, in order to improve the capacity of signal acquisition more effectively, and the selective adoption of flexible optical fiber can realize a compact opto-mechanical structure, which is also conducive to the miniaturization of the device. The experimental results show that this apparatus has a high sensitivity, can be used for the detection and analysis of human nutrition.

  8. Laser-saturated fluorescence measurements in laminar sooting diffusion flames

    Science.gov (United States)

    Wey, Changlie

    1993-01-01

    The hydroxyl radical is known to be one of the most important intermediate species in the combustion processes. The hydroxyl radical has also been considered a dominant oxidizer of soot particles in flames. In this investigation the hydroxyl concentration profiles in sooting diffusion flames were measured by the laser-saturated fluorescence (LSF) method. The temperature distributions in the flames were measured by the two-line LSF technique and by thermocouple. In the sooting region the OH fluorescence was too weak to make accurate temperature measurements. The hydroxyl fluorescence profiles for all four flames presented herein show that the OH fluorescence intensities peaked near the flame front. The OH fluorescence intensity dropped sharply toward the dark region of the flame and continued declining to the sooting region. The OH fluorescence profiles also indicate that the OH fluorescence decreased with increasing height in the flames for all flames investigated. Varying the oxidizer composition resulted in a corresponding variation in the maximum OH concentration and the flame temperature. Furthermore, it appears that the maximum OH concentration for each flame increased with increasing flame temperature.

  9. Sequence-dependent fluorescence of cyanine dyes on microarrays.

    Science.gov (United States)

    Agbavwe, Christy; Somoza, Mark M

    2011-01-01

    Cy3 and Cy5 are among the most commonly used oligonucleotide labeling molecules. Studies of nucleic acid structure and dynamics use these dyes, and they are ubiquitous in microarray experiments. They are sensitive to their environment and have higher quantum yield when bound to DNA. The fluorescent intensity of terminal cyanine dyes is also known to be significantly dependent on the base sequence of the oligonucleotide. We have developed a very precise and high-throughput method to evaluate the sequence dependence of oligonucleotide labeling dyes using microarrays and have applied the method to Cy3 and Cy5. We used light-directed in-situ synthesis of terminally-labeled microarrays to determine the fluorescence intensity of each dye on all 1024 possible 5'-labeled 5-mers. Their intensity is sensitive to all five bases. Their fluorescence is higher with 5' guanines, and adenines in subsequent positions. Cytosine suppresses fluorescence. Intensity falls by half over the range of all 5-mers for Cy3, and two-thirds for Cy5. Labeling with 5'-biotin-streptavidin-Cy3/-Cy5 gives a completely different sequence dependence and greatly reduces fluorescence compared with direct terminal labeling.

  10. Sequence-dependent fluorescence of cyanine dyes on microarrays.

    Directory of Open Access Journals (Sweden)

    Christy Agbavwe

    Full Text Available Cy3 and Cy5 are among the most commonly used oligonucleotide labeling molecules. Studies of nucleic acid structure and dynamics use these dyes, and they are ubiquitous in microarray experiments. They are sensitive to their environment and have higher quantum yield when bound to DNA. The fluorescent intensity of terminal cyanine dyes is also known to be significantly dependent on the base sequence of the oligonucleotide. We have developed a very precise and high-throughput method to evaluate the sequence dependence of oligonucleotide labeling dyes using microarrays and have applied the method to Cy3 and Cy5. We used light-directed in-situ synthesis of terminally-labeled microarrays to determine the fluorescence intensity of each dye on all 1024 possible 5'-labeled 5-mers. Their intensity is sensitive to all five bases. Their fluorescence is higher with 5' guanines, and adenines in subsequent positions. Cytosine suppresses fluorescence. Intensity falls by half over the range of all 5-mers for Cy3, and two-thirds for Cy5. Labeling with 5'-biotin-streptavidin-Cy3/-Cy5 gives a completely different sequence dependence and greatly reduces fluorescence compared with direct terminal labeling.

  11. Multispectral fluorescence imaging of human ovarian and fallopian tube tissue for early-stage cancer detection

    Science.gov (United States)

    Tate, Tyler H.; Baggett, Brenda; Rice, Photini F. S.; Koevary, Jennifer Watson; Orsinger, Gabriel V.; Nymeyer, Ariel C.; Welge, Weston A.; Saboda, Kathylynn; Roe, Denise J.; Hatch, Kenneth D.; Chambers, Setsuko K.; Utzinger, Urs; Barton, Jennifer Kehlet

    2016-05-01

    With early detection, 5-year survival rates for ovarian cancer exceed 90%, yet no effective early screening method exists. Emerging consensus suggests over 50% of the most lethal form of the disease originates in the fallopian tube. Twenty-eight women undergoing oophorectomy or debulking surgery provided informed consent for the use of surgical discard tissue samples for multispectral fluorescence imaging. Using multiple ultraviolet and visible excitation wavelengths and emissions bands, 12 fluorescence and 6 reflectance images of 47 ovarian and 31 fallopian tube tissue samples were recorded. After imaging, each sample was fixed, sectioned, and stained for pathological evaluation. Univariate logistic regression showed cancerous tissue samples had significantly lower intensity than noncancerous tissue for 17 image types. The predictive power of multiple image types was evaluated using multivariate logistic regression (MLR) and quadratic discriminant analysis (QDA). Two MLR models each using two image types had receiver operating characteristic curves with area under the curve exceeding 0.9. QDA determined 56 image type combinations with perfect resubstituting using as few as five image types. Adaption of the system for future in vivo fallopian tube and ovary endoscopic imaging is possible, which may enable sensitive detection of ovarian cancer with no exogenous contrast agents.

  12. Stroboscopic fluorescence lifetime imaging.

    Science.gov (United States)

    Holton, Mark D; Silvestre, Oscar R; Errington, Rachel J; Smith, Paul J; Matthews, Daniel R; Rees, Paul; Summers, Huw D

    2009-03-30

    We report a fluorescence lifetime imaging technique that uses the time integrated response to a periodic optical excitation, eliminating the need for time resolution in detection. A Dirac pulse train of variable period is used to probe the frequency response of the total fluorescence per pulse leading to a frequency roll-off that is dependent on the relaxation rate of the fluorophores. The technique is validated by demonstrating wide-field, realtime, lifetime imaging of the endocytosis of inorganic quantum dots by a cancer cell line. Surface charging of the dots in the intra-cellular environment produces a switch in the fluorescence lifetime from approximately 40 ns to technique offers lifetime based imaging at video rates with standard CCD cameras and has application in probing millisecond cell dynamics and in high throughput imaging assays.

  13. Analysis of green fluorescent protein bioluminescence in vivo and in vitro using a glow discharge

    Science.gov (United States)

    Hernández, L.; Mandujano, L. A.; Cuevas, J.; Reyes, P. G.; Osorio-González, D.

    2015-03-01

    The discovery of fluorescent proteins has been a revolution in cell biology and related sciences because of their many applications, mainly emphasizing their use as cellular markers. The green fluorescent protein (GFP) is one of the most used as it requires no cofactors to generate fluorescence and retains this property into any organism when it is expressed by recombinant DNA techniques, which is a great advantage. In this work, we analyze the emission spectra of recombinant green fluorescent protein in vivo and in vitro exposed to a glow discharge plasma of nitrogen in order to relate electron temperature to fluorescence intensity.

  14. Short-range ordered photonic structures of lamellae-forming diblock copolymers for excitation-regulated fluorescence enhancement

    Science.gov (United States)

    Kim, Se Hee; Kim, Ki-Se; Char, Kookheon; Yoo, Seong Il; Sohn, Byeong-Hyeok

    2016-05-01

    Photonic crystals can be represented by periodic nanostructures with alternating refractive indices, which create artificial stop bands with the appearance of colors. In this regard, nanodomains of block copolymers and the corresponding structural colors have been intensively studied in the past. However, the practical application of photonic crystals of block copolymers has been limited to a large degree because of the presence of large defects and grain boundaries in the nanodomains of block copolymers. The present study focuses on the alternative opportunity of short-range ordered nanodomains of block copolymers for fluorescence enhancement, which also has a direct relevance to the development of fluorescence sensors or detectors. The enhancement mechanism was found to be interconnected with the excitation process rather than the alternation of the decay kinetics. In particular, we demonstrate that randomly oriented, but regular grains of lamellae of polystyrene-block-polyisoprene, PS-b-PI, diblock copolymers and their blend with PS homopolymers can behave as Bragg mirrors to induce multiple reflections of the excitation source inside the photonic structures. This process in turn significantly increases the effective absorption of the given fluorophores inside the polymeric photonic structures to amplify the fluorescence signal.Photonic crystals can be represented by periodic nanostructures with alternating refractive indices, which create artificial stop bands with the appearance of colors. In this regard, nanodomains of block copolymers and the corresponding structural colors have been intensively studied in the past. However, the practical application of photonic crystals of block copolymers has been limited to a large degree because of the presence of large defects and grain boundaries in the nanodomains of block copolymers. The present study focuses on the alternative opportunity of short-range ordered nanodomains of block copolymers for fluorescence

  15. Faster Improvement in Migraine Pain Intensity and Migraine-Related Disability at Early Time Points with AVP-825 (Sumatriptan Nasal Powder Delivery System) versus Oral Sumatriptan: A Comparative Randomized Clinical Trial Across Multiple Attacks from the COMPASS Study.

    Science.gov (United States)

    Lipton, Richard B; McGinley, James S; Shulman, Kenneth J; Wirth, R J; Buse, Dawn C

    2017-09-07

    Fast relief of migraine pain, associated symptoms, and migraine-related disability are priorities in the acute treatment of migraine. Efforts to improve the pharmacokinetic profiles of acute migraine treatments with the aim of providing faster relief include the development of non-oral routes of administration. AVP-825 (ONZETRA(®) Xsail(®) ) is a delivery system containing 22 mg sumatriptan powder that uses a patient's own breath to deliver medication intranasally, targeting the upper posterior nasal cavity beyond the narrow nasal valve, an area lined with vascular mucosa conducive to rapid drug absorption into the systemic circulation. While most studies comparing treatments measure differences in proportions of patients achieving a dichotomous endpoint at fixed time intervals, in this study we compare trajectories of migraine pain and disability over time for AVP-825 versus 100 mg oral sumatriptan tablets. We used data from the COMPASS study (NCT01667679, clinicaltrials.gov), a double-blind, double-dummy, active-comparator, cross-over study of people with a diagnosis of migraine. Participants treated up to five qualifying migraine attacks within 1 hour of onset with either AVP-825 plus placebo tablets or 100 mg oral sumatriptan tablets plus placebo delivery system during the first of two 12-week treatment periods, and then switched treatment sequences to treat up to five more attacks in the second treatment period. Patients recorded ordinal migraine pain intensity and migraine-related disability before dosing (predose), and at 10, 15, 30, 45, 60, 90 and 120 minutes. Three-level ordinal multilevel models accounted for unique data structure (repeated measures nested within attacks for each patient) and tested for treatment differences in migraine pain and migraine-related disability through the first 2 hours of attacks post dose. Among 259 study participants (mean age 40.0 years, 84.6% female, 78.4% white), there was significant between and within person

  16. Nanosecond fluorescence spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Leskovar, B.

    1985-03-01

    This article is a summary of a short course lecture given in conjunction with the 1984 Nuclear Science Symposium. Measuring systems for nanosecond fluorescence spectroscopy using single-photon counting techniques are presented. These involve systems based on relaxation-type spark gap light pulser and synchronously pumped mode-locked dye lasers. Furthermore, typical characteristics and optimization of operating conditions of the critical components responsible for the system time resolution are discussed. A short comparison of the most important deconvolution methods for numerical analysis of experimental data is given particularly with respect to the signal-to-noise ratio of the fluorescence signal. 22 refs., 8 figs.

  17. Chamber catalogues of optical and fluorescent signatures distinguish bioaerosol classes

    Science.gov (United States)

    Hernandez, Mark; Perring, Anne E.; McCabe, Kevin; Kok, Greg; Granger, Gary; Baumgardner, Darrel

    2016-07-01

    Rapid bioaerosol characterization has immediate applications in the military, environmental and public health sectors. Recent technological advances have facilitated single-particle detection of fluorescent aerosol in near real time; this leverages controlled ultraviolet exposures with single or multiple wavelengths, followed by the characterization of associated fluorescence. This type of ultraviolet induced fluorescence has been used to detect airborne microorganisms and their fragments in laboratory studies, and it has been extended to field studies that implicate bioaerosol to compose a substantial fraction of supermicron atmospheric particles. To enhance the information yield that new-generation fluorescence instruments can provide, we report the compilation of a referential aerobiological catalogue including more than 50 pure cultures of common airborne bacteria, fungi and pollens, recovered at water activity equilibrium in a mesoscale chamber (1 m3). This catalogue juxtaposes intrinsic optical properties and select bandwidths of fluorescence emissions, which manifest to clearly distinguish between major classes of airborne microbes and pollens.

  18. Bioaerosol Analysis by Online Fluorescence Detection and Fluorescence Microscopy

    Science.gov (United States)

    Huffman, Alex; Pöhlker, Christopher; Treutlein, Bärbel; Pöschl, Ulrich

    2010-05-01

    Primary biological aerosol particles (PBAPs), including bacteria, spores and pollen, are essential for the spread of organisms and disease in the biosphere, and numerous studies have suggested that they may be important for atmospheric processes, including the formation of clouds and precipitation. The atmospheric abundance and size distribution of PBAPs, however, are largely unknown. At a semi-urban site in Mainz, Germany, we used an ultraviolet aerodynamic particle sizer (UV-APS) to measure fluorescent biological aerosol particles (FBAPs), which can be regarded as viable bioaerosol particles representing a lower limit for the actual abundance of PBAPs. Fluorescence of non-biological aerosol components are likely to influence the measurement results obtained for fine particles (concentration of coarse FBAPs was 3x10-2 cm-3, corresponding to 4% of total coarse particle number [1]. The mean mass concentration of FBAPs was 1 ?g m-3, corresponding to 20% of total coarse particle mass. The FBAP number size distributions exhibited alternating patterns with peaks at various diameters, though a pronounced peak at 3 μm was essentially always observed. This peak is likely due to fungal spores or agglomerated bacteria, and it exhibited a pronounced diel cycle with maximum intensity during early/mid-morning. FBAP peaks around 1.5 μm, 5 μm, and 13 μm were also observed, but less pronounced and less frequent. These may be explained by single bacterial cells, larger fungal spores, and pollen grains, respectively. The observed number concentrations and characteristic sizes of FBAPs are consistent with microscopic, biological and chemical analyses of PBAPs in aerosol filter samples. To our knowledge, however, this is the first study reporting continuous online measurements of bioaerosol particles over several months, a range of characteristic size distribution patterns, and a persistent bioaerosol peak at 3 μm. The measurement results confirm that PBAPs account for a

  19. Fluorescent property of 3-hydroxymethyl imidazo[1,2-a]pyridine and pyrimidine derivatives

    Directory of Open Access Journals (Sweden)

    Velázquez-Olvera Stephania

    2012-08-01

    Full Text Available Abstract Background Imidazo[1,2-a]pyridines and pyrimidines are important organic fluorophores which have been investigated as biomarkers and photochemical sensors. The effect on the luminescent property by substituents in the heterocycle and phenyl rings, have been studied as well. In this investigation, series of 3-hydroxymethyl imidazo[1,2-a]pyridines and pyrimidines were synthesized and evaluated in relation to fluorescence emission, based upon the hypothesis that the hydroxymethyl group may act as an enhancer of fluorescence intensity. Results Compounds of both series emitted light in organic solvents dilutions as well as in acidic and alkaline media. Quantitative fluorescence spectroscopy determined that both fused heterocycles fluoresced more intensely than the parent unsubstituted imidazo[1,2-a]azine fluorophore. In particular, 3-hydroxymethyl imidazo[1,2-a]pyridines fluoresced more intensely than 3-hydroxymethyl imidazo[1,2-a]pyrimidines, the latter emitting blue light at longer wavelengths, whereas the former emitted purple light. Conclusion It was concluded that in most cases the hydroxymethyl moiety did act as an enhancer of the fluorescence intensity, however, a comparison made with the fluorescence emitted by 2-aryl imidazo[1,2-a]azines revealed that in some cases the hydroxymethyl substituent decreased the fluorescence intensity.

  20. The effect of temperature of fluorescence: an animal study

    Science.gov (United States)

    Walsh, Alex; Masters, Bart; Jansen, Duco; Welch, A. J.; Mahadevan-Jansen, Anita

    2010-02-01

    The effect of temperature on the fluorescence of enucleated porcine eyes and rat skin was studied. The fluorescence peak intensity was found to decrease as the tissue temperature increased. A dual-excitation, fiber-based system was used to collect fluorescence and diffuse-reflectance spectra from the samples. A thermal camera was used to determine the temperature of the tissue at the time of fluorescence measurement. The samples were mounted in a saline bath and measurements were made as the tissue temperature was increased from -20°C to 70°C. Results indicate that temperature affects several fluorescence spectra characteristics. The peak height decreased as temperature increased. At temperatures above 60°C, the peak position shifted to lower wavelengths. Heating and cooling experiments of the rat skin demonstrate the recovery of the loss in fluorescence. The diffuse reflectance spectra indicated a change in optical properties past 60°C, but prior to the denaturation temperature for collagen at 57°C, no change in optical properties was observed. Results suggest that the decrease in fluorescence is both a property of fluorescence and a result of altering optical properties.

  1. Fluorescence Talbot microscope using incoherent source

    Science.gov (United States)

    Sun, Yangyang; Pang, Shuo

    2016-08-01

    Fluorescence Talbot microscope is a scalable field-of-view (FOV) imaging platform, which takes advantage of the phase sensitivity of the self-image of a periodic structure. Such a system can maintain the microscopic resolution and extend the FOV for the whole slide (15 mm×15 mm) scanning. Previously reported Talbot fluorescence systems, tabletop and on-chip device alike, rely on the coherence of the illumination source, limiting their potential applications in low-resource setting environment. A more cost-effective setup using a light-emitting diode, which has an area of 4 mm2 and a full width at half maximum of 16 nm in wavelength, is demonstrated. Compared to the illumination that is spatially filtered by a single pinhole, our system has achieved an illumination intensity that is 357 times higher. The reconstructed image quality is comparable to that of a 10× microscope objective. Various samples, such as fluorescent beads, green fluorescence protein-labeled HeLa cells, and a mouse kidney slide, were reconstructed by the system.

  2. Fluorescence self-quenching of tetraphenylporphyrin in liquid medium

    Energy Technology Data Exchange (ETDEWEB)

    Ghosh, Mihir [Integrated Science Education and Research Centre, Siksha-Bhavana, Visva-Bharati, Santiniketan 731235 (India); Nath, Sukhendu [Radiation and Photochemistry Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400085 (India); Hajra, Alakananda [Department of Chemistry, Siksha-Bhavana, Visva-Bharati, Santiniketan 731235 (India); Sinha, Subrata, E-mail: subratasinha67@rediffmail.com [Integrated Science Education and Research Centre, Siksha-Bhavana, Visva-Bharati, Santiniketan 731235 (India)

    2013-09-15

    Self-quenching of the fluorescence emission of tetraphenylporphyrin at high concentrations in toluene at the ambient temperature (300 K) is discussed in detail based on steady state and time-resolved fluorescence measurements. The fluorescence self-quenching is mainly attributed to re-absorption effect and the Förster type resonance energy transfer process (homotransfer). The re-absorption effect is found to deform the fluorescence emission spectra significantly in energy positions as well as relative intensities of different peaks at high concentrations. Nearly ideal fluorescence emission spectra are observed at a concentration ∼10{sup −7} mol/L. Moreover, there is an apparent enhancement of the fluorescence lifetime value of tetraphenylporphyrin in toluene at high concentrations, especially on the blue side of the fluorescence emission spectra. To the best knowledge of the authors, this is the first detail report on the fluorescence self-quenching of porphyrins in liquid medium. This finding carries great importance in view of the widespread research on porphyrins in the fields of solar light harvesting, artificial photosynthesis, photodynamic therapy, etc. -- Highlights: • The effect of concentration on the fluorescence emission spectra of tetraphenylporphyrin (TPhP) in toluene at 300 K is investigated by using steady state and time-resolved fluorescence techniques. • Both re-absorption effect and the Förster type resonance energy transfer are found to be responsible for the observed fluorescence self-quenching at high concentrations. • These investigations are extremely important in view of the extensive applications of porphyrins in the fabrication of molecular electronic devices, especially for solar and artificial photosynthetic devices, where highly concentrated porphyrins are often used for efficient light harvesting.

  3. Construction of fluorescence resonance energy transfer vectors and their application in study of structure and function of signal transducers and activators of transcription 1

    Institute of Scientific and Technical Information of China (English)

    Fujun Han; Yongfeng Luo; Nanhai Ge; Jun Xu

    2008-01-01

    Protein-protein interactions have been studied extensively by green fluorescent protein-based fluorescence resonance energy transfer (FRET). The fluorescent proteins (FP) can be fused either to the N- or C-terminus of a host protein, but it is difficult to predict which order will perturb the host protein the least and provide the largest FRET. Therefore, a researcher needs to fuse host proteins with FP at both the N- and C-termini and test every possible combination (N-N,N-C, or C-C) to promote the energy transfer efficiency.Consequently, researchers required to do many subelonings.Herein, we designed FRET vectors to make them more efficient. The expression vectors ofpCTP.YFP and pYFP-CFP were constructed with both cyan fluorescent protein (CFP)-yellow fluorescent protein (YFP) and YFP-CFP coding sequences flanked by two restriction enzyme sites, and with multiple cloning regions in the middle of both coding sequences. To select an optimal combination for FRET detection, we created plasmids encoding various fusion proteins of FP and signal transducers and activators of transcription 1 (STAT1). We found that the nuclear:cytoplasmic fluorescence intensity ratios of STAT1 -FP were significantly higher than those of FP-STAT1 at steady state,and fluorescence redistribution was only observed for STAT1-FP upon interferon gamma (IFNΥ) stimulation. In addition, positive FRET signals were only detected in the C-C interactions of STAT 1 homodimer. Taken together, these data indicate that fusing STATI at the N.terminus with Fpimpairs the interactions ofunphospborylated STAT1 homodimers and possibly diminishes its binding with DNA. In contrast, STATIFP was functional with respect to its activation. Moreover, the FRET vectors are able to facilitate FRET studies.

  4. 多元智能理论下英语专业精读课程任务型教学模式探索%Exploration on the Mode of Task-based Teaching of Intensive Reading Course for English Majors with the Theory of Multiple Intelligences

    Institute of Scientific and Technical Information of China (English)

    姜冬艳

    2011-01-01

    With the theory of multiple intelligences,this article analyzes the existing problems in current task-based teaching of intensive reading course for English majors,and discussed the advantages and construction of the new mode of task-based teaching with the theory of multiple intelligences.%本文从多元智能理论出发,分析了当前英语专业精读课程任务型教学存在的问题,探讨了融入多元智能理论的英语专业精读课程任务型教学新模式的优势及构建。

  5. Directly labeled fluorescent DNA probes for chromosome mapping

    Energy Technology Data Exchange (ETDEWEB)

    Marrone, B.L.; Deaven, L.L.; Chen, D.J.; Park, Min S.; MacInnes, M.A.; Salzman, G.C.; Yoshida, T.M.

    1995-12-31

    A new strategy is briefly described for employing nucleic acid probes that are directly labeled with fluorochromes in fluorescence in situ hybridization techniques. These probes will permit the detection, quantitation, and high-precision spatial analysis of multiple DNA sequences along a single chromosome using video-enhanced fluorescence microscopy and digital image processing and analysis. Potential advantages of direct labeled DNA probes for fluorescence in situ hybridization far surpass currently available, indirect DNA probe labeling techniques in ease of use, versatility, and increased signal- to-noise ratio.

  6. Fluorescence Experiments with Quinine

    Science.gov (United States)

    O'Reilly, James E.

    1975-01-01

    Describes a series of experiments which illustrate the analytical capabilities of fluorescence, and outlines two straightforward analyses involving real analyses. These experiments are suitable for an undergraduate instrumental analysis course and require approximately six to seven hours of laboratory time. (MLH)

  7. FLEX: fluorescence explorer

    NARCIS (Netherlands)

    Stoll, M.Ph.; Court, A.J.; Smorenburg, C.; Visser, H.; Crocco, L.; Heilimo, J.; Honig, A.

    1999-01-01

    FLEX is a scientifically driven space mission to provide demonstration/validation of the instrumentation and technique for measuring the natural fluorescence of vegetation in the Fraunhofer lines. The payload consists of high spectral resolution (0.1-0.3 nm) CCD imaging grating spectrometer with two

  8. Improved method for efficient imaging of intracellular Cl(-) with Cl-Sensor using conventional fluorescence setup.

    Science.gov (United States)

    Friedel, Perrine; Bregestovski, Piotr; Medina, Igor

    2013-01-01

    Chloride (Cl(-)) homeostasis is known to be fundamental for central nervous system functioning. Alterations in intracellular Cl(-) concentration ([Cl(-)]i) and changes in the efficacy of Cl(-) extrusion are involved in numerous neurological disorders. Therefore, there is a strong need for studies of the dynamics of [Cl(-)]i in different cell types under physiological conditions and during pathology. Several previous works reported having successfully achieved recording of [Cl(-)]i using genetically encoded Cl-Sensor that is composed of the cyan fluorescent protein (CFP) and Cl(-)-sensitive mutant of the yellow fluorescent protein (YFPCl). However, all reported works were performed using specially designed setups with ultra-sensitive CCD cameras. Our multiple attempts to monitor Cl(-)-dependent fluorescence of Cl-Sensor using conventional epifluorescence microscopes did not yield successful results. In the present work, we have analysed the reason of our failures and found that they were caused by a strong inactivation of the YFPCl component of Cl-Sensor during excitation of the CFP with 430 nm light. Based on the obtained results, we reduced 20-fold the intensity of the 430 nm excitation and modified the recording protocol that allows now stable long-lasting ratiometric measurements of Cl-Sensor fluorescence in different cell types including cultured hippocampal neurons and their tiny dendrites and spines. Simultaneous imaging and patch clamp recording revealed that in mature neurons, the novel protocol allows detection of as little as 2 mM changes of [Cl(-)]i from the resting level of 5-10 mM. We demonstrate also a usefulness of the developed [Cl(-)]i measurement procedure for large scale screening of the activity of exogenously expressed potassium-chloride co-transporter KCC2, a major neuronal Cl(-) extruder that is implicated in numerous neurological disorders and is a target for novel therapeutical treatments.

  9. Improved method for efficient imaging of intracellular Cl- with Cl-Sensor using conventional fluorescence setup

    Directory of Open Access Journals (Sweden)

    Perrine eFriedel

    2013-04-01

    Full Text Available Chloride (Cl- homeostasis is known to be fundamental for central nervous system functioning. Alterations in intracellular Cl- concentration ([Cl-]i and changes in the efficacy of Cl- extrusion are involved in numerous neurological disorders. Therefore there is a strong need for studies of the dynamics of [Cl-]i in different cell types under physiological conditions and during pathology. Several previous works reported having successfully achieved recording of [Cl-]i using genetically encoded Cl-Sensor that is composed of the cyan fluorescent protein (CFP and Cl--sensitive mutant of the yellow fluorescent protein (YFPCl. However all reported works were performed using specially designed setups with ultra-sensitive CCD cameras. Our multiple attempts to monitor Cl--dependent fluorescence of Cl-Sensor using conventional epifluorescence microscopes did not yield successful results. In the present work, we have analysed the reason of our failures and found that they were caused by a strong inactivation of the YFPCl component of Cl-Sensor during excitation of the CFP with 430 nm light. Based on the obtained results, we reduced 20-fold the intensity of the 430 nm excitation and modified the recording protocol that allows now stable long-lasting ratiometric measurements of Cl-Sensor fluorescence in different cell types including cultured hippocampal neurons and their tiny dendrites and spines. Simultaneous imaging and patch clamp recording revealed that in mature neurons, the novel protocol allows detection of as little as 2 mM changes of [Cl-]i from the resting level of 5-10 mM. We demonstrate also a usefulness of the developed [Cl-]i measurement procedure for large scale screening of the activity of exogenously expressed potassium-chloride co-transporter KCC2, a major neuronal Cl- extruder, that is implicated in numerous neurological disorders and is a target for novel therapeutical treatments.

  10. Fluorescent dye labeled DNA size standards for molecular mass detection in visible/infrared range

    Directory of Open Access Journals (Sweden)

    Sreelakshmi Yellamaraju

    2011-01-01

    Full Text Available Abstract Background Targeting Induced Local Lesions in Genomes (TILLING is a high throughput reverse genetics tool which detects mismatches (single point mutations or small indels in large number of individuals of mutagenized populations. Currently, TILLING is intensively used for genomics assisted molecular breeding of several crop plants for desired traits. Most commonly used platform for mutation detection is Li-COR DNA Analyzer, where PCR amplified products treated with single strand mismatch specific nuclease are resolved on denaturing gels. The molecular size of any cut product can be easily estimated by comparing with IR dye labeled markers of known sizes. Similar fluorescent dye labeled size markers are also used for several genotyping experiments. Currently, commercially available size standards are expensive and are restricted up to only 700 bp which renders estimation of products of sizes greater than 700 bases inaccurate. Findings A simple protocol was developed for labeling 5' end of multiple DNA size markers with fluorescent dyes. This method involves cloning a pool of different size markers of DNA in a plasmid vector. PCR amplification of plasmid using IR dye labeled universal primers generates 5' fluorescent labeled products of various sizes. The size of products constituting the ladder can be customized as per the need. The generated size markers can be used without any further purification and were found to be stable up to one year at -20°C. Conclusions A simple method was developed for generating fluorescent dye labeled size standards. This method can be customized to generate different size standards as per experimental needs. The protocol described can also be adapted for developing labeled size standards for detection on platforms other than Li-COR i.e. other than infra red range of the spectrum.

  11. Plasmonically amplified bioassay - Total internal reflection fluorescence vs. epifluorescence geometry.

    Science.gov (United States)

    Hageneder, Simone; Bauch, Martin; Dostalek, Jakub

    2016-08-15

    This paper investigates plasmonic amplification in two commonly used optical configurations for fluorescence readout of bioassays - epifluorescence (EPF) and total internal reflection fluorescence (TIRF). The plasmonic amplification in the EPF configuration was implemented by using crossed gold diffraction grating and Kretschmann geometry of attenuated total reflection method (ATR) was employed in the TIRF configuration. Identical assay, surface architecture for analyte capture, and optics for the excitation, collection and detection of emitted fluorescence light intensity were used in both TIRF and EPF configurations. Simulations predict that the crossed gold diffraction grating (EPF) can amplify the fluorescence signal by a factor of 10(2) by the combination of surface plasmon-enhanced excitation and directional surface plasmon-coupled emission in the red part of spectrum. This factor is about order of magnitude higher than that predicted for the Kretschmann geometry (TIRF) which only took advantage of the surface plasmon-enhanced excitation. When applied for the readout of sandwich interleukin 6 (IL-6) immunoassay, the plasmonically amplified EPF geometry designed for Alexa Fluor 647 labels offered 4-times higher fluorescence signal intensity compared to TIRF. Interestingly, both geometries allowed reaching the same detection limit of 0.4pM despite of the difference in the fluorescence signal enhancement. This is attributed to inherently lower background of fluorescence signal for TIRF geometry compared to that for EPF which compensates for the weaker fluorescence signal enhancement. The analysis of the inflammation biomarker IL-6 in serum at medically relevant concentrations and the utilization of plasmonic amplification for the fluorescence measurement of kinetics of surface affinity reactions are demonstrated for both EPF and TIRF readout.

  12. Novel application of low pH-dependent fluorescent dyes to examine colitis

    Directory of Open Access Journals (Sweden)

    Watanabe Osamu

    2010-01-01

    Full Text Available Abstract Background Endoscopy capable of fluorescence observation provides histological information on gastrointestinal lesions. We explored the novel application of low pH-dependent fluorescent dyes for fluorescence observation of crypt structure and inflammatory cell infiltration in the colon. Methods Low pH-dependent fluorescent dyes were applied to the colonic mucosa of normal mice for observation under fluorescence stereomicroscopy system. We also examined mouse models of colitis, which were induced by trinitrobenzenesulfonic acid, dextran sulfate sodium or interleukin-10 deficiency. Results Topical application of low pH-dependent fluorescent dyes revealed crypts as ring-shaped fluorescent stains by visualizing the mucin granules of goblet cells. Because of the minimal fluorescence intensity of the low pH-dependent fluorescent dyes in phosphate-buffered saline, it was not necessary to wash the mucosa before the fluorescence observation. 4-Nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ was quicker to achieve complete staining (three minutes than LysoSensor Green DND-153 and DND-189 (20 minutes. In each type of colitis, NBD-PZ revealed the destruction of the crypts as the disappearance of the ring-shaped fluorescent stains and the infiltration of inflammatory cells as the aggregation of punctate fluorescent stains through visualization of lysosomes. Conclusions Low pH-dependent fluorescent dyes, especially NBD-PZ, are suitable for topical application to the colonic mucosa and have characteristics that allow for the histological examination of colitis.

  13. Highly sensitive turn-on fluorescence detection of thrombomodulin based on fluorescence resonance energy transfer

    Science.gov (United States)

    Kong, Liyan; Zhu, Jiaming; Wang, Wen; Jin, Lehe; Fu, Yanjiao; Duan, Bohui; Tan, Liang

    2017-02-01

    As an integral glycoprotein on the surface of endothelial cells, thrombomodulin (TM) has very high affinity for thrombin. TM has been regarded to be a marker of endothelial damage since it can be released during endothelial cell injury. In this work, a highly sensitive fluorescence method for the quantitative detection of TM was developed. TM antibody (Ab) and bovine serum albumin (BSA) were bound on gold nanoparticles (AuNPs) to construct BSA-AuNPs-Ab nanocomposites and they were characterized by transmission electron microscope and UV-vis spectrophotometry. The fluorescence of acridine orange (AO) was quenched by the prepared gold nanocomposites based on fluorescence resonance energy transfer (FRET). In the presence of TM, the fluorescence was turned on due to the effective separation of AO from the surface of gold nanocomposites. Under optimum conditions, the enhanced fluorescence intensity displayed a linear relationship with the logarithm of the TM concentration from 0.1 pg mL- 1 to 5 ng mL- 1 with a low detection limit of 12 fg mL- 1. The release of soluble thrombomodulin (sTM) by the injured HUVEC-C cells in the presence of H2O2 was investigated using the proposed method. The released sTM content in the growth medium was found to be increased with the enhancement of contact time of the cells with H2O2.

  14. Fluorescence derivatization of clarithromycin for high performance liquid chromatographic determination with fluorescence detection

    Directory of Open Access Journals (Sweden)

    Boonleang, J.

    2007-03-01

    Full Text Available Clarithromycin (CAM is a semisynthetic macrolide antibiotic whose chemical structure has no suitable chromophore for highly sensitive and accurate direct determination. The aim of this study was toderivatize CAM with fluorescence-labeling compounds capable of enhancing the sensitivity of CAM determination. Two fluorescence-labeling compounds were used in this study, 9-fluorenylmethyloxycarbonylchloride (FMOC-Cl and 1-naphthylisocyanate (NIC, both of which gave the fluorescent derivatives of CAM with approximately the same fluorescence intensity. The derivatization reactions in the concentration rangestudied (0.1-2.4 μg/ml were reproducible with coefficient of variation of less than 6.01% and the fluorescence responses were linearly proportional to CAM concentration with r2 of greater than 0.99. The reaction of CAM with FMOC-Cl optimally occurred in a solvent mixture of acetonitrile and phosphate buffer pH 7.5(4:1 volume ratio at 40oC for 40 min. The optimum derivatization reaction of CAM with NIC took place in acetonitrile with triethylamine as catalyst at 30oC for 60 min. It was mild and quantitative giving CAM-NICfluorescent derivative, which is more stable at room temperature than CAM-FMOC derivative. This derivatization should, therefore, be more applicable for highly sensitive CAM determination especially for thestudy involving the analysis of several samples.

  15. Fluorescence lifetime measurements of intrinsically unstructured proteins: application to α-synuclein.

    Science.gov (United States)

    Schreurs, Sarah; Kluba, Malgorzata; Meuvis, Jessika; Engelborghs, Yves

    2012-01-01

    Lifetimes of fluorescent states and their fluorescence intensities are strictly coupled and very sensitive to the environment of the fluorophores. The advantage of measuring lifetimes, next to intensities, comes from the fact that it can reveal heterogeneity and dynamic properties of this environment. In this way lifetime analysis can be used to characterize static and dynamic conformational properties and heterogeneity of fluorescent groups in different areas of a protein and as a function of time for an evolving protein. The phenomena that determine the lifetime of a label are its intrinsic properties, dynamic quenching by neighboring groups, exposure to the solvent, as well as Förster resonance energy transfer (FRET) between different groups. The basic principles of these fluorescence phenomena can be found extensively described in the excellent book of Lakowicz (Principles of fluorescence spectroscopy, 3rd edn. Springer, New York, 2006). The fluorescent groups involved are either natural amino acid side chains like tryptophan (Trp) or tyrosine (Tyr), or fluorescent labels covalently engineered into the protein. Even a single fluorescent group can show indications of heterogeneity in the local environment. If several natural fluorescent groups are present, the properties of the individual groups can be separated using site-directed mutagenesis, and additivity of their contributions can be analyzed (Engelborghs, Spectrochim Acta A Mol Biomol Spectrosc 57(11):2255-2270, 2001). If no fluorescent group is naturally present, site-directed mutagenesis can be used to introduce either a fluorescent amino acid or a cysteine allowing chemical labeling.

  16. Fluorescence and fluorescence-lifetime imaging microscopy (FLIM) to characterize yeast strains by autofluorescence

    Science.gov (United States)

    Bhatta, H.; Goldys, E. M.; Ma, J.

    2006-02-01

    We characterised populations of wild type baking and brewing yeast cells using intrinsic fluorescence and fluorescence lifetime microscopy, in order to obtain quantitative identifiers of different strains. The cell autofluorescence was excited at 405 nm and observed within 440-540 nm range where strong cell to cell variability was observed. The images were analyzed using customised public domain software, which provided information on cell size, intensity and texture-related features. In light of significant diversity of the data, statistical methods were utilized to assess the validity of the proposed quantitative identifiers for strain differentiation. The Kolmogorov-Smirnov test was applied to confirm that empirical distribution functions for size, intensity and entropy for different strains were statistically different. These characteristics were followed with culture age of 24, 48 and 72 h, (the latter corresponding to a stationary growth phase) and size, and to some extent entropy, were found to be independent of age. The fluorescence intensity presented a distinctive evolution with age, different for each of the examined strains. The lifetime analysis revealed a short decay time component of 1.4 ns and a second, longer one with the average value of 3.5 ns and a broad distribution. High variability of lifetime values within cells was observed however a lifetime texture feature in the studied strains was statistically different.

  17. The application of time decay characteristics of laser-induced fluorescence in the classification of vegetation.

    Science.gov (United States)

    Gong, Wei; Yang, Jian; Shi, Shuo; Du, Lin; Sun, Jia; Song, Shalei

    2017-02-01

    In this study, the time decay of the chlorophyll fluorescence intensity (TDCFI) of vegetation was measured based on laser-induced fluorescence (LIF) technology with a 355 nm laser serving as the excitation light source. The pseudo-color diagram of the TDCFI (PDTDCFIs) was proposed for use as a characteristic fingerprint for the analysis of various plant species based on variations in the fluorescence intensity over time. Compared with the steady-state fluorescence spectra, two-dimensional PDTDCFIs contained more spectral information, including variations in both the shape of the laser-induced fluorescence spectra and the relative intensity. The experimental results demonstrated that the PDTDCFIs of various plant species show distinct differences, and this was successfully applied in the classification of plant species. Therefore, the PDTDCFIs of plants could provide researchers with a more reliable and useful tool for the characterization of vegetation. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  18. Portable fluorescence meter for medical applications

    Science.gov (United States)

    Kornilin, Dmitriy V.; Grishanov, Vladimir N.

    2016-04-01

    Recently, there are great deals of skin fluorescence studies for diagnostic purposes in medicine. Measurement of the intensity of autofluorescence (AF) is suitable method for diagnostic, because it does not require traumatic procedures. Skin AF is widely used by doctors in order to assess the concentration of advanced glycation endproduct (AGE). There are no in vivo fluorescence meters made in Russia, which are affordable, portable, easy-to-use and easily replicable. This paper is devoted to study of the fluorimeter and its mathematical model of spectral characteristics that were developed by authors. Fluorimeter and its software are fully operational and they were given to doctors for testing in the real clinic conditions in order to get a set of AF statistics for patients.

  19. Fluorescence Spectroscopic Studies on Ovis Lactoperoxidase

    Directory of Open Access Journals (Sweden)

    P. V. Joseph

    2004-01-01

    Full Text Available Ovis lactoperoxidase (sLP, on excitation at 280 nm shows fluorescence emission of a single broad maximum at 332 nm. The conformational stability was measured by unfolding studies in urea and guanidine hydrochloride. The fluorescence intensity gradually decreased with increase in urea concentrations. The decline might have been caused by partial unfolding, affecting some of the tryptophan residues. In 5 M GuHCl concentrations, a red shift in emission maximum to 356 nm was observed. It indicates that tryptophan is buried in the interior of the hydrophobic environment in native folded state and inaccessible to solvent water but on unfolding all get exposed to aqueous environment. Acrylamide is an efficient quencher and the quenching process is essentially homogenous with all tryptophan being accessible. A little quenching is observed for KI is interpreted as sLP has tryptophan residues that are buried inside the core of the protein.

  20. 1-Imino Nitroxide Pyrene, a Burst-type H+ Fluorescent Switch

    Institute of Scientific and Technical Information of China (English)

    Jun LI; Hong Mei WANG; Hui Min MA; De Qing ZHANG; Shao Xiang XIONG

    2005-01-01

    1-Imino nitroxide pyrene 1 has been characterized as a H+ fluorescent switch. The effects of various interfering species, solvents and the irradiation of ultraviolet light on the fluorescence intensity of 1 are also investigated. 1 fluoresces weakly in the 85% ethanol media containing no more than 4.5×10-5 mol/L of H+ (HCl), but can produce a burst increase of about 4times in the fluorescence intensity when the concentration of H+ is increased to or over 4.9×10-5mol/L. The extremely sharp response of 1 to H+ occurs only within such a narrow concentration range. Moreover, the decrease of H+ concentration through adding sodium hydroxide causes the fluorescence quenching, suggesting that 1 may serve as a fluorescent switch for monitoring the concentration change of H+ around the point of 4.5× 10-5 mol/L.

  1. Fluorescence dynamics of interactions between polyamide PyPyPyβDp and DNA

    Institute of Scientific and Technical Information of China (English)

    ZHANG; Huijuan; WANG; Jin; WU; Yishi; YUAN; Gu; AI; Xicheng; WANG; Li

    2006-01-01

    The photophysical properties of the polyamide PyPyPyβDp (PPP) were investigated by means of steady-state absorption and fluorescence spectroscopies, as well as time-resolved fluorescence spectroscopy. It was found that the excited-state properties of PPP are very sensitive to solvents. In TKMC buffer PPP exhibited weak fluorescence with a decay time constant of 16 ps, while with the decrease of the solvent polarity PPP showed the blue-shifted peak position, increased intensity and lengthened life-time for its fluorescence behavior. In the presence of calf thymus DNA, it was observed that the fluorescence intensity was enhanced and the fluorescence lifetime increased from 16 to 32 ps for PPP, which verified that PPP bound into the minor groove of DNA duplex.

  2. Tracking quasi-stationary flow of weak fluorescent signals by adaptive multi-frame correlation.

    Science.gov (United States)

    Ji, L; Danuser, G

    2005-12-01

    We have developed a novel cross-correlation technique to probe quasi-stationary flow of fluorescent signals in live cells at a spatial resolution that is close to single particle tracking. By correlating image blocks between pairs of consecutive frames and integrating their correlation scores over multiple frame pairs, uncertainty in identifying a globally significant maximum in the correlation score function has been greatly reduced as compared with conventional correlation-based tracking using the signal of only two consecutive frames. This approach proves robust and very effective in analysing images with a weak, noise-perturbed signal contrast where texture characteristics cannot be matched between only a pair of frames. It can also be applied to images that lack prominent features that could be utilized for particle tracking or feature-based template matching. Furthermore, owing to the integration of correlation scores over multiple frames, the method can handle signals with substantial frame-to-frame intensity variation where conventional correlation-based tracking fails. We tested the performance of the method by tracking polymer flow in actin and microtubule cytoskeleton structures labelled at various fluorophore densities providing imagery with a broad range of signal modulation and noise. In applications to fluorescent speckle microscopy (FSM), where the fluorophore density is sufficiently low to reveal patterns of discrete fluorescent marks referred to as speckles, we combined the multi-frame correlation approach proposed above with particle tracking. This hybrid approach allowed us to follow single speckles robustly in areas of high speckle density and fast flow, where previously published FSM analysis methods were unsuccessful. Thus, we can now probe cytoskeleton polymer dynamics in living cells at an entirely new level of complexity and with unprecedented detail.

  3. Management of fluorescent lamps in controlled environment chambers

    Science.gov (United States)

    Romer, Mark

    1994-01-01

    Management of fluorescent lights is recommended to (1) maintain uniformity of light intensity over time and (2) permit reproducibility of lighting conditions during experimental replications. At the McGill Phytotron, the lighting intensity can be controlled to desired level because any individual pair of the 40 lamps in each chamber can be set to be 'on' at any particular time. A lamp canopy service history is maintained for each experiment permitting accurate replication of lighting conditions for subsequent replicate trials.

  4. Position-specific incorporation of fluorescent non-natural amino acids into maltose-binding protein for detection of ligand binding by FRET and fluorescence quenching.

    Science.gov (United States)

    Iijima, Issei; Hohsaka, Takahiro

    2009-04-17

    Position-specific incorporation of fluorescent groups is a useful method for analysis of the functions and structures of proteins. We have developed a method for the incorporation of visible-wavelength-fluorescent non-natural amino acids into proteins in a cell-free translation system. Using this technique, we introduced one or two BODIPY-linked amino acids into maltose-binding protein (MBP) to obtain MBP derivatives showing ligand-dependent changes in fluorescence intensity or intensity ratio. BODIPY-FL-aminophenylalanine was incorporated in place of 15 tyrosines, as well as the N-terminal Lys1, and the C-terminal Lys370 of MBP. Fluorescence measurements revealed that MBP containing a BODIPY-FL moiety in place of Tyr210 showed a 13-fold increase in fluorescence upon binding of maltose. Tryptophan-to-phenylalanine substitutions suggest that the increase in fluorescence was the result of a decrease in the quenching of BODIPY-FL by tryptophan located around the binding site. MBP containing a BODIPY-558 moiety also showed a maltose-dependent increase in fluorescence. BODIPY-FL was then additionally incorporated in place of Lys1 of the BODIPY-558-containing MBP as a response to the amber codon. Fluorescence measurements with excitation of BODIPY-FL showed a large change in fluorescence intensity ratio (0.13 to 1.25) upon binding of maltose; this change can be attributed to fluorescence resonance energy transfer (FRET) and maltose-dependent quenching of BODIPY-558. These results demonstrate the usefulness of the position-specific incorporation of fluorescent amino acids in the fluorescence-based detection of protein functions.

  5. Fluorescence quantum yields of natural organic matter and organic compounds: Implications for the fluorescence-based interpretation of organic matter composition

    DEFF Research Database (Denmark)

    Wünsch, Urban; Murphy, Kathleen R.; Stedmon, Colin

    2015-01-01

    Absorbance and fluorescence spectroscopy are economical tools for tracing the supply, turnover and fate of dissolved organic matter (DOM). The colored and fluorescent fractions of DOM (CDOM and FDOM, respectively) are linked by the apparent fluorescence quantum yield (AQY) of DOM, which reflects ...... to confirm matches was limited due to multiple compounds exhibiting very similar spectra. This reiterates the fact that spectral similarity alone is insufficient evidence of the presence of particular compounds, and additional evidence is required...

  6. Progressive increase of T1 signal intensity in the dentate nucleus and globus pallidus on unenhanced T1-weighted MR images in the pediatric brain exposed to multiple doses of gadolinium contrast.

    Science.gov (United States)

    Roberts, Donna R; Holden, Kenton R

    2016-03-01

    Recently, there have been reports of gadolinium accumulation in the brain and bone of adult patients with normal renal function who have undergone multiple gadolinium contrast administrations. This case report gives the first description of a pediatric patient who, following multiple contrasted MRI exams, demonstrated abnormal signal on unenhanced T1-weighted imaging involving the dentate nucleus and globus pallidus, a finding which has previously been shown to represent gadolinium deposition in adults. The patient presented here had no history of intracranial pathology which would alter the blood brain barrier or abnormal renal function. The clinical significance of gadolinium accumulation in the human body is currently unknown but is of concern, particularly in pediatric patients who have a lifetime to manifest any potential adverse consequences. Therefore, research is needed to address the clinical significance, if any, of gadolinium deposition in the developing pediatric brain. Given these current uncertainties, clinicians should continue to use prudence in selecting pediatric patients to undergo contrasted MRI and in selecting the appropriate contrast agents to use.

  7. Magnetic-fluorescent-targeting multifunctional aptasensorfor highly sensitive and one-step rapid detection of ochratoxin A.

    Science.gov (United States)

    Wang, Chengquan; Qian, Jing; Wang, Kan; Wang, Kun; Liu, Qian; Dong, Xiaoya; Wang, Chengke; Huang, Xingyi

    2015-06-15

    A multifunctional aptasensor for highly sensitive and one-step rapid detection of ochratoxin A (OTA), has been developed using aptamer-conjugated magnetic beads (MBs) as the recognition and concentration element and a heavy CdTe quantum dots (QDs) as the label. Initially, the thiolated aptamer was conjugated on the Fe3O4@Au MBs through Au-S covalent binding. Subsequently, multiple CdTe QDs were loaded both in and on a versatile SiO2 nanocarrier to produce a large amplification factor of hybrid fluorescent nanoparticles (HFNPs) labeled complementary DNA (cDNA). The magnetic-fluorescent-targeting multifunctional aptasensor was thus fabricated by immobilizing the HFNPs onto MBs' surface through the hybrid reaction between the aptamer and cDNA. This aptasensor can be produced at large scale in a single run, and then can be conveniently used for rapid detection of OTA through a one-step incubation procedure. The presence of OTA would trigger aptamer-OTA binding, resulting in the partial release of the HFNPs into bulk solution. After a simple magnetic separation, the supernatant liquid of the above solution contained a great number of CdTe QDs produced an intense fluorescence emission. Under the optimal conditions, the fluorescence intensity of the released HFNPs was proportional to the concentration of OTA in a wide range of 15 pg mL(-1) -100 ng mL(-1) with a detection limit of 5.4 pg mL(-1) (S/N=3). This multifunctional aptasensor represents a promising path toward routine quality control of food safety, and also creates the opportunity to develop aptasensors for other targets using this strategy.

  8. Spatial distribution of the internal and near-field intensities of large cylindrical and spherical scatterers

    Science.gov (United States)

    Benincasa, Daniel S.; Barber, Peter W.; Zhang, Jian-Zhi; Hsieh, Wen-Feng; Chang, Richard K.

    1987-04-01

    Spatial distributions of the near-field and internal electromagnetic intensities have been calculated and experimentally observed for dielectric cylinders and spheres which are large relative to the incident wavelength. Two prominent features of the calculated results are the high intensity peaks which exist in both the internal and near fields of these objects, even for nonresonant conditions, and the well-defined shadow behind the objects. Such intensity distributions were confirmed by using the fluorescence from iodine vapor to image the near-field intensity distribution and the fluorescence from ethanol droplets impregnated with rhodamine 590 to image the internal-intensity distribution.

  9. Emotionally Intense Science Activities

    Science.gov (United States)

    King, Donna; Ritchie, Stephen; Sandhu, Maryam; Henderson, Senka

    2015-08-01

    Science activities that evoke positive emotional responses make a difference to students' emotional experience of science. In this study, we explored 8th Grade students' discrete emotions expressed during science activities in a unit on Energy. Multiple data sources including classroom videos, interviews and emotion diaries completed at the end of each lesson were analysed to identify individual student's emotions. Results from two representative students are presented as case studies. Using a theoretical perspective drawn from theories of emotions founded in sociology, two assertions emerged. First, during the demonstration activity, students experienced the emotions of wonder and surprise; second, during a laboratory activity, students experienced the intense positive emotions of happiness/joy. Characteristics of these activities that contributed to students' positive experiences are highlighted. The study found that choosing activities that evoked strong positive emotional experiences, focused students' attention on the phenomenon they were learning, and the activities were recalled positively. Furthermore, such positive experiences may contribute to students' interest and engagement in science and longer term memorability. Finally, implications for science teachers and pre-service teacher education are suggested.

  10. Portable fluorescence meter with reference backscattering channel

    Science.gov (United States)

    Kornilin, Dmitriy V.; Grishanov, Vladimir N.; Zakharov, Valery P.; Burkov, Dmitriy S.

    2016-09-01

    Methods based on fluorescence and backscattering are intensively used for determination of the advanced glycation end products (AGE) concentration in the biological tissues. There are strong correlation between the AGE concentration and the severity of such diseases like diabetes, coronary heart disease and renal failure. This fact can be used for diagnostic purposes in medical applications. Only few investigations in this area can be useful for development of portable and affordable in vivo AGE meter because the most of them are oriented on using spectrometers. In this study we describe the design and the results of tests on volunteers of portable fluorescence meter based on two photodiodes. One channel of such fluorimeter is used for measurement of the autofluorescence (AF) intensity, another one - for the intensity of elastically scattered radiation, which can be used as a reference. This reference channel is proposed for normalization of the skin autofluorescence signal to the human skin photo type. The fluorimeter, that was developed is relatively compact and does not contain any expensive optical and electronic components. The experimental results prove that proposed tool can be used for the AGE estimation in human skin.

  11. Fluorescence spectroscopic detection of early injury-induced atherosclerosis

    Science.gov (United States)

    Lucas, Alexandra; Perk, Masis; Wen, Yue; Smith, Carol

    1992-08-01

    Laser-induced fluorescence spectroscopy has been used for the detection of advanced atherosclerotic lesions. Angioplasty balloon-mediated injury was examined spectroscopically in order to assess the sensitivity of fluorescence spectroscopy for detection of early atherosclerosis. Abdominal aortic balloon angioplasty was performed via femoral artery cutdown in nine White Leghorn roosters (five normal, four atherogenic diet). Roosters were sacrificed at 1, 2, 4, 8, and 12 week intervals. Fluorescence emission spectra (n equals 114) were recorded from each aortic section (XeCl excimer laser, 308 nm, 1.5 - 2.0 mJ/pulse, 5 Hz). Changes in normalized fluorescence emission intensity were correlated with selected sections of histology. All balloon-injured segments showed intimal fibrous proliferation. For intimal thickness measuring > 70 (mu) , fluorescence emission intensity was decreased at 440 - 460 nm (p Lesions complicated by thrombus also had lower fluorescence emission at 425 - 450 nm when compared to histologically normal aorta (p muscular (abdominal) aorta (p muscular abdominal aorta.

  12. Fluorescence spectrum analysis of atherosclerotic plaque using doxycycline.

    Science.gov (United States)

    Miyagi, M; Nakajima, H; Katoh, T; Usui, M; Amemiya, T; Nagai, Y; Ibukiyama, C

    1999-05-01

    Using doxycycline (DOXY), fluorescence spectrum analysis was performed on arteriosclerotic lesions, and the efficacy of this method was examined in basic and clinical studies. In the basic study, DOXY 50 mg was administered intravenously to arteriosclerotic rabbits, and the thoracoabdominal aorta removed. Fluorescence spectral analysis was performed on each specimen, and the fluorescence spectral pattern, peak intensity and degree of intimal hypertrophy were studied. In the clinical study, DOXY 200 mg was administered intravenously to 6 human subjects with stable angina and coronary arterial stenosis of greater than 90%, and coronary angiography, coronary angioscopy and fluorescence spectral analysis were performed. DOXY accumulation in the arteriosclerotic intima of rabbit aortae was confirmed. The fluorescence spectrum was monomodal, peaking at around 532 nm. In the noncalcification group, significant correlation was observed between peak intensity and arteriosclerotic intimal thickness. Using DOXY as a fluorescent marker, it was possible to assess the level of arteriosclerotic intimal hypertrophy. Clinically, it was possible to obtain the DOXY spectrum of the coronary arteries.

  13. Laser-stimulated fluorescence in paleontology.

    Science.gov (United States)

    Kaye, Thomas G; Falk, Amanda R; Pittman, Michael; Sereno, Paul C; Martin, Larry D; Burnham, David A; Gong, Enpu; Xu, Xing; Wang, Yinan

    2015-01-01

    Fluorescence using ultraviolet (UV) light has seen increased use as a tool in paleontology over the last decade. Laser-stimulated fluorescence (LSF) is a next generation technique that is emerging as a way to fluoresce paleontological specimens that remain dark under typical UV. A laser's ability to concentrate very high flux rates both at the macroscopic and microscopic levels results in specimens fluorescing in ways a standard UV bulb cannot induce. Presented here are five paleontological case histories that illustrate the technique across a broad range of specimens and scales. Novel uses such as back-lighting opaque specimens to reveal detail and detection of specimens completely obscured by matrix are highlighted in these examples. The recent cost reductions in medium-power short wavelength lasers and use of standard photographic filters has now made this technique widely accessible to researchers. This technology has the potential to automate multiple aspects of paleontology, including preparation and sorting of microfossils. This represents a highly cost-effective way to address paleontology's preparatory bottleneck.

  14. Laser-stimulated fluorescence in paleontology.

    Directory of Open Access Journals (Sweden)

    Thomas G Kaye

    Full Text Available Fluorescence using ultraviolet (UV light has seen increased use as a tool in paleontology over the last decade. Laser-stimulated fluorescence (LSF is a next generation technique that is emerging as a way to fluoresce paleontological specimens that remain dark under typical UV. A laser's ability to concentrate very high flux rates both at the macroscopic and microscopic levels results in specimens fluorescing in ways a standard UV bulb cannot induce. Presented here are five paleontological case histories that illustrate the technique across a broad range of specimens and scales. Novel uses such as back-lighting opaque specimens to reveal detail and detection of specimens completely obscured by matrix are highlighted in these examples. The recent cost reductions in medium-power short wavelength lasers and use of standard photographic filters has now made this technique widely accessible to researchers. This technology has the potential to automate multiple aspects of paleontology, including preparation and sorting of microfossils. This represents a highly cost-effective way to address paleontology's preparatory bottleneck.

  15. Polarisation Sensitive Single Molecule Fluorescence Detection with Linear Polarised Excitation Light and Modulated Polarisation Direction Applied to Multichromophoric Entities

    NARCIS (Netherlands)

    Gensch, T.; Hofkens, J.; Köhn, F.; Vosch, T.; Herrmann, A.; Müllen, K.; Schryver, F.C. De

    2001-01-01

    Recently, investigations of the fluorescence properties of a multichromophoric dendritic entity at the single molecule level have revealed multiple fluorescence levels, collective off-states, variations of the polarisation, large shifts in the spectral position and changes in the fluorescence decay

  16. Rise-time of FRET-acceptor fluorescence tracks protein folding

    OpenAIRE

    Simon Lindhoud; Adrie H. Westphal; van Mierlo, Carlo P. M.; Visser, Antonie J. W. G.; Jan Willem Borst

    2014-01-01

    Uniform labeling of proteins with fluorescent donor and acceptor dyes with an equimolar ratio is paramount for accurate determination of Förster resonance energy transfer (FRET) efficiencies. In practice, however, the labeled protein population contains donor-labeled molecules that have no corresponding acceptor. These FRET-inactive donors contaminate the donor fluorescence signal, which leads to underestimation of FRET efficiencies in conventional fluorescence intensity and lifetime-based F...

  17. Determination of elements in some lichens growing in Giresun and Ordu province (Turkey) using energy dispersive X-ray fluorescence spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Aslan, A. [Department of Biology, Education Faculty, Atatuerk University, 25240 Erzurum (Turkey); Budak, G. [Department of Physics, Science and Art Faculty, Atatuerk University, 25240 Erzurum (Turkey)]. E-mail: gbudak@atauni.edu.tr; Tirasoglu, E. [Department of Physics, Faculty of Arts and Sciences, Karadeniz Technical University, Trabzon (Turkey); Karabulut, A. [Department of Physics, Science and Art Faculty, Atatuerk University, 25240 Erzurum (Turkey)

    2006-01-15

    The concentration of five different elements in six lichens species of different regions in Giresun and Ordu (Turkey) was determined using the energy dispersive X-ray fluorescence method. A radioisotope excited X-ray fluorescence analysis using the method of multiple standard addition is applied to the elemental analysis of lichens. An annular 50 mCi {sup 241}Am radioactive source and an annular 50 mCi {sup 55}Fe radioactive source were used for excitation of characteristic K X-rays. An Si(Li)detector which has a 147 eV full-width at half-maximum for 5.9 keV photons was used for intensity measurements. A qualitative analysis of spectral peaks showed that the samples contained potassium, calcium, titanium, iron, and barium.

  18. Burdekin River Runoff Reconstruction from Fluorescence Data in Havannah and Pandora Reefs for 1644 to 1980

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Annual (water year, October-September) runoff (mm) of the Burdekin River, Queensland, Australia was reconstructed from intensity of fluorescence measured in two...

  19. Fluorescence nanoscopy. Methods and applications

    OpenAIRE

    Requejo-Isidro, Jose

    2013-01-01

    Fluorescence nanoscopy refers to the experimental techniques and analytical methods used for fluorescence imaging at a resolution higher than conventional, diffraction-limited, microscopy. This review explains the concepts behind fluorescence nanoscopy and focuses on the latest and promising developments in acquisition techniques, labelling strategies to obtain highly detailed super-resolved images and in the quantitative methods to extract meaningful information from them.

  20. Photoacoustic-fluorescence in vitro flow cytometry for quantification of absorption, scattering and fluorescence properties of the cells

    Science.gov (United States)

    Nedosekin, D. A.; Sarimollaoglu, M.; Foster, S.; Galanzha, E. I.; Zharov, V. P.

    2013-03-01

    Fluorescence flow cytometry is a well-established analytical tool that provides quantification of multiple biological parameters of cells at molecular levels, including their functional states, morphology, composition, proliferation, and protein expression. However, only the fluorescence and scattering parameters of the cells or labels are available for detection. Cell pigmentation, presence of non-fluorescent dyes or nanoparticles cannot be reliably quantified. Herewith, we present a novel photoacoustic (PA) flow cytometry design for simple integration of absorbance measurements into schematics of conventional in vitro flow cytometers. The integrated system allow simultaneous measurements of light absorbance, scattering and of multicolor fluorescence from single cells in the flow at rates up to 2 m/s. We compared various combinations of excitation laser sources for multicolor detection, including simultaneous excitation of PA and fluorescence using a single 500 kHz pulsed nanosecond laser. Multichannel detection scheme allows simultaneous detection of up to 8 labels, including 4 fluorescent tags and 4 PA colors. In vitro PA-fluorescence flow cytometer was used for studies of nanoparticles uptake and for the analysis of cell line pigmentation, including genetically encoded melanin expression in breast cancer cell line. We demonstrate that this system can be used for direct nanotoxicity studies with simultaneous quantification of nanoparticles content and assessment of cell viability using a conventional fluorescent apoptosis assays.

  1. Fluorescence characteristics of polychlorinated biphenyl isomers in cyclodextrin media

    Energy Technology Data Exchange (ETDEWEB)

    Femia, R.A.; Scypinski, S.; Love, L.J.C.

    1985-01-01

    The fluorescence characteristics of several polychlorinated biphenyl (PCB) isomers in ..cap alpha..- and ..beta..-cyclodextrin (CD) are discussed and contrasted. The steric hindrance of PCBs imposed by the positions of the chlorine atoms on the rings determines the overall stability of the resulting inclusion complexes with cyclodextrin which is reflected in the fluorescence intensities. Less substituted homologues include into the CD cavity equally well in both cyclodextrins, but very heavily substituted molecules show drastic differences between their fluorescence intensities in the ..cap alpha..- and ..beta..-cyclodextrin solutions. This behavior can be employed as a method for spectral fractionation of PCB isomers. Spectral separation is demonstrated here for two variably substituted molecules, and the photophysical limitations of this approach are discussed.

  2. Quantitative analysis for nonlinear fluorescent spectra based on edges matching

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A novel spectra-edge-matching approach is proposed for the quantitative analysis of the nonlinear fluorescence spectra of the air impurities excited by a femtosecond laser.The fluorescence spectra are first denoised and compressed,both by wavelet transform,and several peak groups are then picked from each spectrum according to a threshold of intensity and are used to extract the spectral features through principal component analysis.It is indicated that the first two principle components actually cover up to 98% of the total information and are sufficient for the final concentration analysis.The analysis reveals a monotone relationship between the spectra intensity and the concentration of the air impurities,suggesting that the femtosecond laser induced fluorescence spectroscopy along with the proposed spectra analysis method can become a powerful tool for monitoring environmental pollutants.

  3. Background suppression in fluorescence nanoscopy with stimulated emission double depletion

    Science.gov (United States)

    Gao, Peng; Prunsche, Benedikt; Zhou, Lu; Nienhaus, Karin; Nienhaus, G. Ulrich

    2017-01-01

    Stimulated emission depletion (STED) fluorescence nanoscopy is a powerful super-resolution imaging technique based on the confinement of fluorescence emission to the central subregion of an observation volume through de-excitation of fluorophores in the periphery via stimulated emission. Here, we introduce stimulated emission double depletion (STEDD) as a method to selectively remove artificial background intensity. In this approach, a first, conventional STED pulse is followed by a second, delayed Gaussian STED pulse that specifically depletes the central region, thus leaving only background. Thanks to time-resolved detection we can remove this background intensity voxel by voxel by taking the weighted difference of photons collected before and after the second STED pulse. STEDD thus yields background-suppressed super-resolved images as well as STED-based fluorescence correlation spectroscopy data. Furthermore, the proposed method is also beneficial when considering lower-power, less redshifted depletion pulses.

  4. Measurement of cell volume changes by fluorescence self-quenching

    DEFF Research Database (Denmark)

    Hamann, Steffen; Kiilgaard, J.F.; Litman, Thomas

    2002-01-01

    At high concentrations, certain fluorophores undergo self-quenching, i.e., fluorescence intensity decreases with increasing fluorophore concentration. Accordingly, the self-quenching properties can be used for measuring water volume changes in lipid vesicles. In cells, quantitative determination...... of water transport using fluorescence self-quenching has been complicated by the requirement of relatively high (mM) and often toxic loading concentrations. Here we report a simple method that uses low (muM) loading concentrations of calcein-acetoxymethyl ester (calcein-AM) to obtain intracellular...... concentrations of the fluorophore calcein suitable for measurement of changes in cell water volume by self-quenching. The relationship between calcein fluorescence intensity, when excited at 490 nm (its excitation maximum), and calcein concentration was investigated in vitro and in various cultured cell types...

  5. Laser induced fluorescence measurements of the mixing of fuel oil with air

    Energy Technology Data Exchange (ETDEWEB)

    Arnold, A.; Bombach, R.; Hubschmid, W.; Kaeppeli, B. [Paul Scherrer Inst. (PSI), Villigen (Switzerland)

    1999-08-01

    We report on measurements of the mixing of fuel oil with air at atmospheric pressure in an industrial premixed gas turbine burner. The concentration of the vaporized fuel oil was measured with laser induced fluorescence. We reason that the fuel oil concentration can be considered with good accuracy as proportional to the fluorescence intensity. (author) 6 fig., 3 refs.

  6. Multiple Sclerosis

    Science.gov (United States)

    Multiple sclerosis (MS) is a nervous system disease that affects your brain and spinal cord. It damages the ... attacks healthy cells in your body by mistake. Multiple sclerosis affects women more than men. It often begins ...

  7. Multiple Myeloma

    Science.gov (United States)

    Multiple myeloma is a cancer that begins in plasma cells, a type of white blood cell. These cells ... bones. No one knows the exact causes of multiple myeloma, but it is more common in older people ...

  8. Theory of analytical curves in atomic fluorescence flame spectrometry

    NARCIS (Netherlands)

    Hooymayers, H.P.

    1968-01-01

    An explicit expression for the intensity of atomic resonance fluorescence as a function of atomic concentration in a flame is derived under certain idealized conditions. The expression is generally valid for a pure Doppler absorption line profile as well as for a combined Doppler and collisional bro

  9. [Synthesis of functionalized cyanines. Fluorescence properties following complexation of cations].

    Science.gov (United States)

    Mazières, M R; Duprat, C; Sutra, E; Lamandé, L; Bergon, M; Bellan, J; Wolf, J G; Roques, C

    2003-01-01

    The ionophoric properties of podands containing dioxazaphosphocane moieties linked by inactive spacers were studied. To increase the detection sensibility of these compounds we introduced a cyanine as spacer. Fluorescence analysis demonstrated the interest of cyanines as active spacers since the complexation by cations as Ca2+ and Mg2+ gives an enhancement of the emission intensity.

  10. The Effect of Time on Bone Fluorescence: Implications for Using Alternate Light Sources to Search for Skeletal Remains.

    Science.gov (United States)

    Swaraldahab, Mohamed A H; Christensen, Angi M

    2016-03-01

    Bones fluoresce when exposed to certain wavelengths of shortwave light, and this property can be useful in locating and sorting skeletal remains in forensic contexts. The proteins in bone collagen are largely responsible for its fluorescent properties, but these proteins degrade and denature over time. This study examined the fluorescence of bones from four temporal groups (recent, semi-recent, ancient, and historic) ranging from 0 to 1064 years before present. Specimens were photographed under 490 nm wavelength light, and fluorescence was quantified by converting intensity to a gray scale value based on the RGB color model using ImageJ(®) software. Significant (p fluorescence between all four temporal groups, and a 0.324 coefficient of correlation indicates a significant (inverse) relationship between fluorescence and time. Bone fluorescence decreases with time, but some fluorescence is retained even in older samples. Fluorescence can therefore be reliably used in many modern skeletal remains searches.

  11. "Color Timer" mice: visualization of neuronal differentiation with fluorescent proteins.

    Science.gov (United States)

    Kanki, Hiroaki; Shimabukuro, Marilia Kimie; Miyawaki, Atsushi; Okano, Hideyuki

    2010-02-02

    The molecular mechanisms governing the differentiation of neural stem cells (NSCs) into neuronal progenitor cells and finally into neurons are gradually being revealed. The lack of convenient means for real-time determination of the stages of differentiation of individual neural cells, however, has been hindering progress in elucidating the mechanisms. In order to be able to easily identify the stages of differentiation of neural cells, we have been attempting to establish a mouse system that would allow progression of neuronal differentiation to be visualized based on transitions between fluorescence colors by using a combination of mouse genetics and the ever-expanding repertoire of fluorescent proteins. In this study we report the initial version of such a mouse system, which we call "Color Timer." We first generated transgenic (Tg; nestin/KOr Tg) mice in which production of the fluorescent protein Kusabira-Orange (KOr) is controlled by the gene regulatory elements within the 2nd intronic enhancer of the nestin gene, which is a good marker for NSCs, so that NSCs would emit orange fluorescence upon excitation. We then confirmed by immunohistochemical and immunocytochemical analyses that the KOr fluorescence closely reflected the presence of the Nestin protein. We also confirmed by a neurosphere formation assay that the intensity of the KOr fluorescence correlated with "stemness" of the cell and it was possible to readily identify NSCs in the two neurogenic regions, namely the dentate gyrus of the hippocampus and the subventricular zone of the lateral ventricle, in the brain of adult nestin/KOr Tg mice by the orange fluorescence they emitted. We then crossed nestin/KOr mice with doublecortin-enhanced Green Fluorescent Protein Tg mice, whose immature neurons emit green fluorescence upon excitation, and it was possible to visualize the progress of NSC-to-neuron differentiation by the transition between fluorescence colors from orange to green. This two-color initial

  12. Iowa Intensive Archaeological Survey

    Data.gov (United States)

    Iowa State University GIS Support and Research Facility — This shape file contains intensive level archaeological survey areas for the state of Iowa. All intensive Phase I surveys that are submitted to the State Historic...

  13. Measuring Physical Activity Intensity

    Medline Plus

    Full Text Available ... for a breath. Absolute Intensity The amount of energy used by the body per minute of activity. ... or vigorous-intensity based upon the amount of energy used by the body while doing the activity. ...

  14. Measuring Physical Activity Intensity

    Medline Plus

    Full Text Available ... Older Adults Overcoming Barriers Measuring Physical Activity Intensity Target Heart Rate & Estimated Maximum Heart Rate Perceived Exertion ( ... a heavy backpack Other Methods of Measuring Intensity Target Heart Rate and Estimated Maximum Heart Rate Perceived ...

  15. Graphene as a Reversible and Spectrally Selective Fluorescence Quencher

    Science.gov (United States)

    Salihoglu, Omer; Kakenov, Nurbek; Balci, Osman; Balci, Sinan; Kocabas, Coskun

    2016-09-01

    We report reversible and spectrally selective fluorescence quenching of quantum dots (QDs) placed in close proximity to graphene. Controlling interband electronic transitions of graphene via electrostatic gating greatly modifies the fluorescence lifetime and intensity of nearby QDs via blocking of the nonradiative energy transfer between QDs and graphene. Using ionic liquid (IL) based electrolyte gating, we are able to control Fermi energy of graphene in the order of 1 eV, which yields electrically controllable fluorescence quenching of QDs in the visible spectrum. Indeed, our technique enables us to perform voltage controllable spectral selectivity among quantum dots at different emission wavelengths. We anticipate that our technique will provide tunable light-matter interaction and energy transfer that could yield hybrid QDs-graphene based optoelectronic devices with novel functionalities, and additionally, may be useful as a spectroscopic ruler, for example, in bioimaging and biomolecular sensing. We propose that graphene can be used as an electrically tunable and wavelength selective fluorescence quencher.

  16. Intraoperative fluorescence diagnosis for removal of cervical and thoracic ependymoma

    Directory of Open Access Journals (Sweden)

    A. M. Zaytcev

    2014-01-01

    Full Text Available The case of successful intraoperative fluorescence diagnosis (IOFD for removal of cervical and thoracic ependymoma performed in P.A. Herzen MCRI is reported. For FD we used the Alasens (Research Institute of Organic Semi-Finished Products and Dyes. The drug solution was given per os at a dose of 20 mg/kg body weight 2.5 h before surgery. IOFD was per-formed 3 h after intake of photosensitizer. For fluorescence diagnosis there was average in-tensity of fluorescence in tumor and no fluorescence in normal spinal tissues. The extent of surgery was determined according to results of IOFD. The control MRI of cervical and supeior thoracic spine with contrast enhancement and follow-up confirmed definitive removal of tumor and showed no postoperative complications.

  17. Fluorescence cell imaging and manipulation using conventional halogen lamp microscopy.

    Directory of Open Access Journals (Sweden)

    Kazuo Yamagata

    Full Text Available Technologies for vitally labeling cells with fluorescent dyes have advanced remarkably. However, to excite fluorescent dyes currently requires powerful illumination, which can cause phototoxic damage to the cells and increases the cost of microscopy. We have developed a filter system to excite fluorescent dyes using a conventional transmission microscope equipped with a halogen lamp. This method allows us to observe previously invisible cell organelles, such as the metaphase spindle of oocytes, without causing phototoxicity. Cells remain healthy even after intensive manipulation under fluorescence observation, such as during bovine, porcine and mouse somatic cell cloning using nuclear transfer. This method does not require expensive epifluorescence equipment and so could help to reduce the science gap between developed and developing countries.

  18. Absolute calibration of the Auger fluorescence detectors

    Energy Technology Data Exchange (ETDEWEB)

    Bauleo, P.; Brack, J.; Garrard, L.; Harton, J.; Knapik, R.; Meyhandan, R.; Rovero, A.C.; /Buenos Aires, IAFE; Tamashiro, A.; Warner, D.

    2005-07-01

    Absolute calibration of the Pierre Auger Observatory fluorescence detectors uses a light source at the telescope aperture. The technique accounts for the combined effects of all detector components in a single measurement. The calibrated 2.5 m diameter light source fills the aperture, providing uniform illumination to each pixel. The known flux from the light source and the response of the acquisition system give the required calibration for each pixel. In the lab, light source uniformity is studied using CCD images and the intensity is measured relative to NIST-calibrated photodiodes. Overall uncertainties are presently 12%, and are dominated by systematics.

  19. Rainfed intensive crop systems

    DEFF Research Database (Denmark)

    Olesen, Jørgen E

    2014-01-01

    This chapter focuses on the importance of intensive cropping systems in contributing to the world supply of food and feed. The impact of climate change on intensive crop production systems is also discussed.......This chapter focuses on the importance of intensive cropping systems in contributing to the world supply of food and feed. The impact of climate change on intensive crop production systems is also discussed....

  20. Quantitative Fluorescence Studies in Living Cells: Extending Fluorescence Fluctuation Spectroscopy to Peripheral Membrane Proteins

    Science.gov (United States)

    Smith, Elizabeth Myhra

    The interactions of peripheral membrane proteins with both membrane lipids and proteins are vital for many cellular processes including membrane trafficking, cellular signaling, and cell growth/regulation. Building accurate biophysical models of these processes requires quantitative characterization of the behavior of peripheral membrane proteins, yet methods to quantify their interactions inside living cells are very limited. Because peripheral membrane proteins usually exist both in membrane-bound and cytoplasmic forms, the separation of these two populations is a key challenge. This thesis aims at addressing this challenge by extending fluorescence fluctuation spectroscopy (FFS) to simultaneously measure the oligomeric state of peripheral membrane proteins in the cytoplasm and at the plasma membrane. We developed a new method based on z-scan FFS that accounts for the fluorescence contributions from cytoplasmic and membrane layers by incorporating a fluorescence intensity z-scan through the cell. H-Ras-EGFP served as a model system to demonstrate the feasibility of the technique. The resolvability and stability of z-scanning was determined as well as the oligomeric state of H-Ras-EGFP at the plasma membrane and in the cytoplasm. Further, we successfully characterized the binding affinity of a variety of proteins to the plasma membrane by quantitative analysis of the z-scan fluorescence intensity profile. This analysis method, which we refer to as z-scan fluorescence profile deconvoution, was further used in combination with dual-color competition studies to determine the lipid specificity of protein binding. Finally, we applied z-scan FFS to provide insight into the early assembly steps of the HTLV-1 retrovirus.

  1. Plasmonic-enhanced two-photon fluorescence with single gold nanoshell

    Science.gov (United States)

    Zhang, TianYue; Lu, GuoWei; Shen, HongMing; Perriat, P.; Martini, M.; Tillement, O.; Gong, QiHuang

    2014-06-01

    Single gold nanoshell with mutilpolar plasmon resonances is proposed to enhance two-photon fluorescence efficiently. The single emitter single nanoshell configuration is studied systematically by employing the finite-difference time-domain method. The emitter located inside or outside the nanoshell at various positions leads to a significantly different enhancement effect. The fluorescent emitter placed outside the nanoshell can achieve large fluorescence intensity given that both the position and orientation of the emission dipole are optimally controlled. In contrast, for the case of the emitter placed inside the nanoshell, it can experience substantial two-photon fluorescence enhancement without strict requirements upon the position and dipole orientations. Metallic nanoshell encapsulating many fluorescent emitters should be a promising nanocomposite configuration for bright two-photon fluorescence label. The results provide a comprehensive understanding about the plasmonic-enhanced two-photon fluorescence behaviors, and the nanocomposite configuration has great potential for optical detecting, imaging and sensing in biological applications.

  2. Rise-Time of FRET-Acceptor Fluorescence Tracks Protein Folding

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    Lindhoud, Simon; Westphal, Adrie H.; van Mierlo, Carlo P. M.; Visser, Antonie J. W. G.; Borst, Jan Willem

    2014-01-01

    Uniform labeling of proteins with fluorescent donor and acceptor dyes with an equimolar ratio is paramount for accurate determination of Förster resonance energy transfer (FRET) efficiencies. In practice, however, the labeled protein population contains donor-labeled molecules that have no corresponding acceptor. These FRET-inactive donors contaminate the donor fluorescence signal, which leads to underestimation of FRET efficiencies in conventional fluorescence intensity and lifetime-based FRET experiments. Such contamination is avoided if FRET efficiencies are extracted from the rise time of acceptor fluorescence upon donor excitation. The reciprocal value of the rise time of acceptor fluorescence is equal to the decay rate of the FRET-active donor fluorescence. Here, we have determined rise times of sensitized acceptor fluorescence to study the folding of double-labeled apoflavodoxin molecules and show that this approach tracks the characteristics of apoflavodoxinʼs complex folding pathway. PMID:25535076

  3. Plasmonic Molecular Nanohybrids—Spectral Dependence of Fluorescence Quenching

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    Maria Olejnik

    2012-01-01

    Full Text Available We demonstrate strong spectral dependence of the efficiency of fluorescence quenching in molecular systems composed of organic dyes and gold nanoparticles. In order to probe the coupling with metallic nanoparticles we use dyes with varied spectral overlap between the plasmon resonance and their absorption. Hybrid molecular structures were obtained via conjugation of metallic nanoparticles with the dyes using biotin-streptavidin linkage. For dyes featuring absorption above the plasmon excitation in gold nanoparticles, laser excitation induces minute changes in the fluorescence intensity and its lifetime for both conjugated and non-conjugated mixtures, which are the reference. In contrast, when the absorption of the dye overlaps with the plasmon resonance, the effect is quite dramatic, reaching 85% and 95% fluorescence quenching for non-conjugated and conjugated mixtures, respectively. The degree of fluorescence quenching strongly depends upon the concentration of metallic nanoparticles. Importantly, the origin of the fluorescence quenching is different in the case of the conjugated mixture, as evidenced by time-resolved fluorescence. For conjugated mixtures of dyes resonant with plasmon, excitation features two-exponential decay. This is in contrast to the single exponential decay measured for the off-resonant configuration. The results provide valuable insight into spectral dependence of the fluorescence quenching in molecular assemblies involving organic dyes and metallic nanoparticles.

  4. Fluorescence of Alexa fluor dye tracks protein folding.

    Science.gov (United States)

    Lindhoud, Simon; Westphal, Adrie H; Visser, Antonie J W G; Borst, Jan Willem; van Mierlo, Carlo P M

    2012-01-01

    Fluorescence spectroscopy is an important tool for the characterization of protein folding. Often, a protein is labeled with appropriate fluorescent donor and acceptor probes and folding-induced changes in Förster Resonance Energy Transfer (FRET) are monitored. However, conformational changes of the protein potentially affect fluorescence properties of both probes, thereby profoundly complicating interpretation of FRET data. In this study, we assess the effects protein folding has on fluorescence properties of Alexa Fluor 488 (A488), which is commonly used as FRET donor. Here, A488 is covalently attached to Cys69 of apoflavodoxin from Azotobacter vinelandii. Although coupling of A488 slightly destabilizes apoflavodoxin, the three-state folding of this protein, which involves a molten globule intermediate, is unaffected. Upon folding of apoflavodoxin, fluorescence emission intensity of A488 changes significantly. To illuminate the molecular sources of this alteration, we applied steady state and t